The primary purpose of this book and its companion volume The Neuropharmacology of Nicotine Dependence is to explore the ways in which recent studies on nicotine and its role in tobacco addiction have opened our eyes to the psychopharmacological properties of this unique and fascinating drug. While the present volume considers the molecular and genetic factors which influence behavioral responses to nicotine and how these may impact on the role of nicotine in tobacco dependence, the book The Neuropharmacology of Nicotine Dependence focuses on the complex neural and psychological mechanisms that mediate nicotine dependence in experimental animal models and their relationship to tobacco addiction in humans.  These volumes will provide readers with a contemporary overview of current research on nicotine psychopharmacology and its role in tobacco dependence from leaders in this field of research and will hopefully prove valuable to those who are developing their own research programmes in this important topic.

Only in recent times has the possibility of growing and implanting replacement teeth, made from one's own cells, moved into the realm of realistic possibilities; however, the molecular and cellular mechanisms of tooth development must be studied in a range of vertebrates, from zebrafish to mice, so that evolutionarily conserved network kernels, which will define the cellular states of generic vertebrate tooth development, can be recognized. In Odontogenesis: Methods and Protocols, experts in the field examine techniques to approach this burgeoning field. This detailed volume includes chapters on the detection of "tooth development" gene expression, both at the RNA and protein level, current approaches to the manipulation of gene expression levels and subsequent analysis of tooth phenotypes, as well as chapters concerning current efforts to get living tooth implants working without waiting for a full understanding of the developmental pathways at the molecular level. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls. Practical and easy to use, Odontogenesis: Methods and Protocols aims to help researchers move forward toward the ultimate goal of getting a "bioengineered tooth" into the patient's mouth.

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 (TM) 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.

This book presents the statistical aspects of designing, analyzing and interpreting the results of genome-wide association scans (GWAS studies) for genetic causes of disease using unrelated subjects. Particular detail is given to the practical aspects of employing the bioinformatics and data handling methods necessary to prepare data for statistical analysis. The goal in writing this book is to give statisticians, epidemiologists, and students in these fields the tools to design a powerful genome-wide study based on current technology. The other part of this is showing readers how to conduct analysis of the created study. Design and Analysis of Genome-Wide Association Studies provides a compendium of well-established statistical methods based upon single SNP associations. It also provides an introduction to more advanced statistical methods and issues. Knowing that technology, for instance large scale SNP arrays, is quickly changing, this text has significant lessons for future use with sequencing data. Emphasis on statistical concepts that apply to the problem of finding disease associations irrespective of the technology ensures its future applications. The author includes current bioinformatics tools while outlining the tools that will be required for use with extensive databases from future large scale sequencing projects. The author includes current bioinformatics tools while outlining additional issues and needs arising from the extensive databases from future large scale sequencing projects.

In Functional Analysis of DNA and Chromatin, expert researchers in the field provide an overview of standard and more recent methods for the functional analysis of the genetic material. These include methods on DNA-dye binding modes, chromatin staining, nuclear dispersion assays, DNA labeling in vivo, sister chromatid exchanges, FISH, DNA and chromatin imaging by fluorescence, electron and atomic force microscopy, detection of apoptotic DNA, cytosine methylation and hydroxy-methylation, DNA thermophoresis, improved methods for histone analysis, chromatin immunoprecipitation, and analysis of rDNA genes and chromatin-associated RNA. 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, Functional Analysis of DNA and Chromatin seeks to aid scientists in the further study of cellular and molecular biology of the nucleus.

Chromatin is of central importance to gene regulation in eukaryotes. Reflecting this singular role for chromatin, numerous approaches have evolved in the laboratory over the past three decades to study chromatin structure and its alterations. Methods of investigating chromatin remodeling, whether in changes in nucleosome structure or position with respect to the incorporated DNA or in histone modifications, have progressed rapidly over the recent years. In Chromatin Remodeling: Methods and Protocols, expert researchers contribute chapters which include methods for investigating chromatin remodeling in vitro and in vivo, in yeast, plants, and mammalian cells, and at local and global levels. Both gene-specific and genome-wide approaches are covered, and in recognition of the increasing prevalence of the latter type of study, the final two chapters focus on bioinformatic/computational approaches to analyzing genome-wide data on chromatin structure. Written in the highly successful Methods in Molecular Biology (TM) series format, the 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. Comprehensive and essential, Chromatin Remodeling: Methods and Protocols serves as a key source of clear techniques as well as a collection of chapters that can inspire future techniques in this vital field of study.

Focusing on in vitro and intracellular RNA structure formation, RNA Folding: Methods and Protocols provides a comprehensive collection of experimental protocols which are suitable to dissect RNA folding pathways and to characterize the structure of RNA folding intermediates at nucleotide or even atomic resolution. The presented techniques include powerful tools with a long tradition in RNA research as well as more advanced, novel methods, thus the methods span multiple disciplines, including molecular biology, biochemistry, biophysics, and computational biology. 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. Practical and authoritative, RNA Folding: Methods and Protocols serves as a vital reference for researchers attempting to gain insights into the secrets of this astounding macromolecule.

This volume provides updates of this established field in both methods and applications, as well as advances in applications of the microarray method to biomarkers such as DNAs, RNAs, proteins, glycans and whole cells. Written for the Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Microarray Technology: Methods and Applications aims to ensure successful results in the further study of this vital field.

The huge potential for gene therapy to cure a wide range of diseases has led to high expectations and a great increase in research efforts in this area, particularly in the study of delivery via viral vectors, widely considered to be more efficient than DNA transfection. In Viral Vectors for Gene Therapy: Methods and Protocols, experts in the field present a collection of their knowledge and experience featuring methodologies that involve virus production, transferring protocols, and evaluating the efficacy of gene products. While thoroughly covering the most popular viral vector systems of adenovirus, retrovirus, and adeno-associated virus, this detailed volume also explores less common viral vector systems such as baculovirus, herpes virus, and measles virus, the growing interest in which is creating a considerable demand for large scale manufacturing and purification procedures. Written in the highly successful Methods in Molecular Biology (TM) series format, many chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and vital tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Viral Vectors for Gene Therapy: Methods and Protocols provides basic principles accessible to scientists from a wide variety of backgrounds for the development of gene therapy viral products that are safe and effective.

PIWI-interacting RNAs (piRNAs) are the third and most-recently discovered group of silencing-inducible small RNAs in animals. PIWI-Interacting RNAs: Methods and Protocols provides the most recent methods and protocols for studying piRNAs in the gonads of a wide range of species, as well as in any other organs where piRNAs may be detected. Comprehensive high-throughput sequencing analysis of piRNAs in embryos, testes and ovaries of D. melanogaster, as well as in mouse and rat testes, has raised the profile of piRNAs and thus further accelerated piRNA studies. In addition to C. elegans, other model species such as Drosophila melanogaster, Arabidopsis thaliana and mice, along with cultured cell lines such as HeLa and Drosophila Schneider 2 (S2), and other such organisms have been used to address the fundamental questions of the biogenesis and functions of RNAi-triggering small non-coding RNAs. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, PIWI-Interacting RNAs: Methods and Protocols seeks to serve both established researchers and newcomers to the field to progress towards the ultimate goal of understanding the mechanisms and actions of piRNAs.

Primary Aldosteronism (PA) is a disease caused by the overproduction of aldosterone hormone from the adrenal glands. PA causes hypertension and the majority with this disease are undiagnosed for PA. There are new insights into this matter by using biochemistry as well as advanced radiology. In 2011, a breakthrough in the genetic derangements came, identifying a mutated potassium channel gene - KCNJ5 - in about 40% of PA with adenoma. Chapters in this book include a history of the disorder, epidemiology, genetics derangements, the KCNJ5 mutations and phenotype and more.

Recent work has revealed that stabilizing G-quadruplexes in telomeric DNA inhibits telomerase activity, providing impetus for the development of G-quartet-interacting drugs, while G-quartet-containing oligonucleotides have been recognized as a potent class of aptamers effective against STAT3 and other transcription factors implicated in oncogenesis, proving these guanine-quartets to be a vital and rich area for future study. In "G-Quadruplex DNA: Methods and Protocols", experts in the field present a collection of detailed techniques for studying G-quartet formation, dynamics, and molecular recognition. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, "G-Quadruplex DNA: Methods and Protocols "promises to be a useful resource for those familiar with G-quartets as well as an easy entry point for those researchers from diverse fields who are just developing an interest in the exciting implications of G-quadruplex DNA.

Carbohydrate microarrays emerged as a key technology for the deciphering of the glycospace by providing a multiplex technology where tens to hundreds of carbohydrates/protein interactions can be probed in parallel. Carbohydrate Microarrays: Methods and Protocols aims to give the reader the theoretical and experimental clues necessary for the fabrication and implementation of carbohydrate microarrays. This requires three essential steps: 1) to obtain the carbohydrate probes (monosacharides, oligosacchrides, polysacchairdes, glycoconjugates or glycoclusters), 2) to immobilize these probes, and 3) to implement the protocols for biological/biochemical interaction with the desired target. This volume gives an overview of carbohydrate microarray and carbohydrate chemistry and illustrates different detection techniques and their applications. 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, Carbohydrate Microarrays: Methods and Protocols compiles a catalogue of protocols on carbohydrate microarrays to span the needs of researchers around the globe.

This volume provides a clear and detailed roadmap of how to design and execute a gene therapy experiment in order to obtain consistent results. Chapters in this book disseminate bits of unknown information that are important to consider during the course of experimentation and will answer questions such as: What delivery vehicle do you use?; How will you ensure that your vector retains stability?; What expression system best fits your needs?; What route will you choose to deliver your gene therapy agent?; How will you model the neurodegenerative disorder that you aim to investigate and what are the proven methods to treat these disorders in preclinical models? 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 thorough, Gene Therapy for Neurological Disorders: Methods and Protocols, is a compilation of protocols and instructive chapters intended to give researchers, clinicians, and students of all levels, a foundation upon which future gene therapy experiments can be designed.

In his 1894 book, Materials for the Study of Variation, William Bateson coined the term Homoeosis with the following prose: The case of the modification of the antenna of an insect into a foot, of the eye of a Crustacean into an antenna, of a petal into a stamen, and the like, are examples of the same kind. It is desirable and indeed necessary that such Variations, which consist in the assumption by one member of a Meristic series, of the form or characters proper to other members of the series, should be recognized as constituting a distinct group of phenomena. ...I therefore propose...the term HOMOEOSIS...; for the essential phenomenon is not that there has merely been a change, but that something has been changed into the likeness of something else. The book was intended as a listing of the kinds of naturally occurring variation that could act as a substrate for the evolutionary process and Bateson took his examples from collections, both private and in museums, of materials displaying morphological oddities. Interestingly the person who also coined the term "Genetics" proffered little in the way of speculation on the possible genetic underpinnings of these oddities. It wasn't until the early part of the next century that these changes in meristic series were shown to be heritable.

During the 20th century Xenopus has flourished as an accessible experimental model system for studying vertebrate biology. The second edition of Xenopus Protocols:Post-Genomic Approaches expands upon the previous edition with novel approaches inspired by Xenopus tropicalis and the sequencing of its genome. Xenopus tropicalis genomics and tractable genetics combined with the unique experimental advantages of Xenopus laevis and their distinctive phylogenetic position make them unique model systems for vertebrate embryonic development in the 21st century. 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, Xenopus Protocols: Post-Genomic Approaches, Second Edition provides a toolkit that will enable every laboratory to maximize the power of this extraordinary experimental system in the post-genomic age.

Chemical genomics technology has been steadily improving, delivering new biological probes and drugs, and the explicit use of the term 'chemical proteomics' has increased with it, as proteins have always been at the heart of this technology. In Chemical Genomics and Proteomics: Reviews and Protocols, experts in the field present updated reviews of the chemistry of small molecules and their interaction with protein targets as well as detailed protocols that cover different types of ligands, carbohydrates, and lipids. For example, the generation of their protein targets and methods for measuring their interactions is covered. Written in the highly successful Methods in Molecular Biology (TM) series format, methodology 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. Thorough and up to date, Chemical Genomics and Proteomics: Reviews and Protocols aims to provide inspiration to those who wish to use chemical genomics and proteomics in their work and develop this young field into full maturity through the incorporation of the new biological and chemical technologies beginning to emerge here.

The determination of protein function has been a major goal of molecular biology since the founding of the discipline. However, as we learn more about gene function, we discover that the context within which a gene is expressed controls the specific function of that gene. It has become critical to establish the background in which gene function is determined and to perform experiments in multiple applicable backgrounds. In Gene Function Analysis, Second Edition, a number of computational and experimental techniques are presented for identifying not only the function of an individual gene, but also the partners that work with that gene. The theme of data integration runs strongly through the computational techniques, with many focusing on gathering data from different sources and different biomolecular types. Experimental techniques have evolved to determine function in specific tissues and at specific times during development. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, Gene Function Analysis, Second Edition seeks to serve both professionals and novices with a growing understanding of the complexity of gene function.

The discovery of RNA interference (RNAi) as a methodology for gene silencing has revolutionized biological research, providing an invaluable avenue for therapeutics, and small interfering RNA (siRNA) is the most common strategy utilized for enacting RNAi. siRNA Design: Methods and Protocols offers expertly crafted guidelines and protocols for the selection of siRNA targeting sequences, for the strategic incorporation of chemical modifications, and for advantageous structural modifications to the classic siRNA design. Protocols are provided for using endogenous cellular machinery to produce siRNA from optimized precursor short hairpin RNA (shRNA) and artificial microRNA (amiRNA) molecules. Strategies are also described for specific applications such as immunostimulatory siRNA that may provide therapeutic benefit against viral infections in mammals, the simultaneous targeting of multiple siRNAs, and siRNA-mediated crop virus resistance. The design of RNAi for gene silencing in embryonic, invertebrate, and plant systems requires a variety of unique approaches, several of which are described towards the end of this volume. Written for the highly successful Methods in Molecular Biology (TM) series, this work contains the kind of detailed description and implementation advice that guarantees successful results. Authoritative and easy to use, siRNA Design: Methods and Protocols will provide researchers, educators, clinicians, and biotech specialists with a broad understanding of the issues in RNAi and how they can be overcome strategically through design.

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.

Since the publication of the popular first edition, the explosion of DNA sequence information, the access to bioinformatics and mutation databases coupled with the ability to readily detect and confirm mutations has cemented the role of molecular diagnostics in medicine and, in particular, mutation detection by the polymerase chain reaction (PCR). In PCR Mutation Detection Protocols, Second Edition, expert researchers bring the subject up-to-date with key protocols involving the PCR and its many various incarnations such as SSCP, CSGE, and dHPLC. The volume also addresses key areas such as Southern blotting, accurate diagnostics with high throughput, as well as microarray systems. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include brief introductions their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes which provide the often hard to find information that may mean the difference between the success and failure of the method. Authoritative and cutting-edge, PCR Mutation Detection Protocols, Second Edition aims to stimulate postgraduate scientists, researchers, and clinicians already engaged in the area and to provide an important first step for those new to this practice wanting to adopt the powerful and essential technique in their own laboratories.

The tiny microRNAs (miRNAs) can have huge impacts on the regulation of a variety of genes and play crucial roles in the fundamental cellular processes. Recent miRNA studies change the landscape of cancer genetics by scrutinizing the alterations of genome-wide miRNA expressions in most common cancers and their regulatory functions during the development of cancer. The connections between miRNAs and cancer are widespread enough to warrant more comprehensive investigations in the systems biology perspective. In MicroRNA and Cancer: Methods and Protocols, internationally renowned experts provide the latest miRNA knowledge, the various techniques and methodologies currently available for cancer research application. Ranging from the fundamental concepts to practical applications, this book presents: * Overview of microRNA biogenesis, computational prediction of new miRNAs in the cancer genome, and miRNA-based therapeutic approaches for cancer treatment * Detailed experimental protocols in miRNA detection with novel and high-throughput technology, miRNA library cloning, miRNA epigenetic regulation, and miRNA pathway study * Stepwise computational and bioinformatic procedures for miRNA complex networks in cancer genomes with a variety of softwares and programs * Cross-cited notes on troubleshooting and avoiding known pitfalls Authoritative and cutting-edge, MicroRNA and Cancer: Methods and Protocols serves researchers with the basic principles of experimental and computational methods for microRNA study in cancer research and provides a firm grounding for those who wish to further develop their own applications and tailor them to their own specific research needs.

microRNAs (miRNAs) are small non-coding RNAs that regulate various biological phenomena, such as development and homeostasis. The dysregulation of miRNA leads to disease progression, particularly of cancer. In Circulating MicroRNAs: Methods and Protocols, expert researchers in the field detail recent advances in the isolation, purification and analysis of circulating miRNAs from a variety of sources for research. The book is divided into three main topics. The first section involves the study of secretory miRNAs in cell-cell communication, and the second, the study of circulating miRNAs in body fluids. The last describes the novel techniques used to study circulating miRNAs. 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, Circulating MicroRNAs: Methods and Protocols seeks to aid scientists in dealing with the recent advances of RNAi technology from the bench to the bedside.

Fluorescence in situ Hybridization (FISH) belongs to that special category of well-established molecular biology techniques that, since their inception a few decades ago, have succeeded in keeping a prominent position within the constantly expanding list of laboratory pro- dures for biomedical research and clinical diagnostics. The design simplicity and cost-effectiveness of the early FISH protocols, combined with the signifcant acceleration of discoveries in related technical areas such as fuor- cence microscopy, digital imaging, and nucleic acid technology have prompted the div- sifcation of the original technique into an outstanding number of imaginative and useful applications, and thus have not only held back its outmoding but have also promoted its expansion into different areas of basic and applied research in the post-genomic era. The 34 chapters included in this book aim at portraying the vibrant complexity and diversity of the current FISH protocol landscape, providing cutting-edge examples of va- ous applications for genetic and developmental research, cancer research, reproductive medicine, diagnostic and prognostic purposes, microbial ecology, and evolutionary st- ies. The book is divided in four parts: (I) Core Techniques, (II) Technical Advancements and Novel Adaptations, (III) Translational FISH: Applications for Human Genetics and Medicine, and (IV) Protocols for Model Organisms.

Small molecule microarrays (SMM) were introduced just a decade ago in 1999 and, within a short space of time, have already established themselves as a vibrant, next generation platform for high-throughput screening. Small Molecule Microarrays: Methods and Protocols showcases a collection of contributions guiding researchers toward ways in which small molecule microarray technology may be deployed for multiplexed screening and profiling. Organized by the categories of small molecules presented on the microarrays, this detailed volume describes in-depth techniques for chemical libraries, peptide libraries, and carbohydrate microarrays. Written in the highly successful Methods in Molecular Biology (TM) series, chapters contain brief introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, vital tips on troubleshooting this often difficult technology, and advice on avoiding known pitfalls. Authoritative and cutting-edge, Small Molecule Microarrays: Methods and Protocols provides meticulous depictions of key hands-on experience and seeks to inspire a future generation of microarray practitioners to take this significant technology forward.