This detailed volume focuses on the CRISPR-associated guide RNA and how it can be designed, modified, and validated for a broad repertoire of purposes. Beginning with a section on computational design of target-specific guide RNAs, the book continues by covering chemical modifications to alter guide RNA stability, specificity, and efficiency, as well as to create inducible guide RNAs, append additional functional domains, and express guide RNAs in a conditional manner. It concludes with methods for measuring off-target guide RNA activity. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and essential, CRISPR Guide RNA Design: Methods and Protocols provides a comprehensive pipeline for guide RNA design and aims to be an invaluable resource in applying this powerful technology to basic research and therapeutic applications.

This detailed edition reflects the significant new findings in the components of permeability barriers and how they work in different tissues with a collection of cutting-edge techniques. Chapters explore the formation, maintenance, regulation, and dynamics of permeability barriers in an effort to push the boundaries of the field. Written for the highly successful 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 up-to-date, Permeability Barrier: Methods and Protocols, Second Edition serves as an invaluable guide for both experts but novices in the stem cell field and other related areas of research.

This book explores the potential applications of animal stem cells in veterinary medicine. It begins with an overview of stem cells and their application in treating various animal diseases, including mastitis. In turn, the book discusses the challenges of using stem cells in regenerative medicine and emphasizes the importance of understanding the action of stem cells and preclinical evidence for ensuring safety and therapeutic efficacy. It also presents methods for the identification, characterization, and quantification of stem cells. Further, it discusses the therapeutic applications of different stem cells, including milk-derived, testicular, and mesenchymal stem cells in veterinary medicine. Lastly, it discusses strategies for and therapeutic applications of genome editing by CRISPER/Cas9 in mammary stem cells. As such, the book offers a valuable resource for students and scientists working in the veterinary sciences and veterinarians.

The reproduction and spread of a virus during an epidemic proceeds when the virus attaches to a host cell and viral genetic material (VGM) (protein, DNA, RNA) enters the cell, then replicates, and perhaps mutates, in the cell. The movement of the VGM across the host cell outer membrane and within the host cell is a spatiotemporal dynamic process that is modeled in this book as a system of ordinary and partial differential equations (ODE/PDEs). The movement of the virus proteins through the cell membrane is modeled as a diffusion process expressed by the diffusion PDE (Fick's second law). Within the cell, the time variation of the VGM is modeled as ODEs. The evolution of the dependent variables is computed by the numerical integration of the ODE/PDEs starting from zero initial conditions (ICs). The departure of the dependent variables from zero is in response to the virus protein concentration at the outer membrane surface (the point at which the virus binds to the host cell). The numerical integration of the ODE/PDEs is performed with routines coded (programmed) in R, a quality, open-source scientific computing system that is readily available from the Internet. Formal mathematics is minimized, e.g., no theorems and proofs. Rather, the presentation is through detailed examples that the reader/researcher/analyst can execute on modest computers. The ODE/PDE dependent variables are displayed graphically with basic R plotting utilities. The R routines are available from a download link so that the example models can be executed without having to first study numerical methods and computer coding. The routines can then be applied to variations and extensions of the ODE/PDE model, such as changes in the parameters and the form of the model equations.

This volume provides detailed methods and key approaches used to mechanistically study DNA damage, as well as the factors involved in the damage response. Chapters guide readers through proteomics and biophysical approaches, analyzing protein function, quantifying DNA replication dynamics and nucleic acid base damage, as well as biochemical reconstitution of key pathways involved in DNA repair. 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, DNA Damage Responses: Methods and Protocols aims to be a useful practical guide to researches to help further their study in this field.

Codon-based models of evolution are a relatively new addition to the toolkit of computational biologists, and in recent years remarkable progress has been made in this area. The study of evolution at the codon level captures information contained in both amino acid and synonymous DNA substitutions. By combining these two types of information, codon analyses are more powerful than those of either amino acid or DNA evolution alone. This is a clear benefit for most evolutionary analyses, including phylogenetic reconstruction, detection of selection, ancestral sequence reconstruction, and alignment of coding DNA. Despite the theoretical advantages of codon based models, their relative complexity delayed their widespread use. Only in recent years, when large-scale sequencing projects produced sufficient genomic data and computational power increased, did their usage become more common. In Codon Evolution, leading researchers in the field of molecular evolution provide the latest insights from codon-based analyses of genetic sequences. The first part of the book provides comprehensive coverage of the developments of various types of codon substitution models such as parametric and empirical models used in maximum likelihood as well as Bayesian frameworks. Subsequent chapters examine the use of codon models to infer selection and other applications of codon models to biological systems. The second part of the book focuses on codon usage bias. Both the underlying mechanisms as well as current methods to analyse codon usage bias are presented.

This book highlights the uses for underutilized crops, presenting the state-of-the-art in terms of genome sequencing for over 30 crops, previously understudied and under-researched. In a changing climate and with significant pressure on the land, it is the ideal time to be discussing novel crops, with significant biotic and abiotic tolerances and/or rich nutrient profiles for consumers. Previously, the only species with sequenced genomes were high-profile internationally recognized crops, but in the current era genomes are being sequenced for dozens of crops, including those previously classified as underutilized, now being investigated. This book covers food crops, from fruits to tubers, and from grasses to legumes, as well as crops with non-food applications. Some of these crops have draft genomes, and others have polished genomes with extensive resequencing panels. Each chapter tells the story of an individual crop or crop group, written by experts, focusing on the genome data available, revealing more about crop domestication and genetic variation, and the current and future prospects given that this data is now becoming available. It also highlights how even small sequencing projects can provide draft genome sequences suitable for gene discovery, comparative genomics, and identification of molecular markers for understanding these crops further.

This detailed book serves as a systematic examination of the analytical methods to study the transcription factor NF- B in physiology and disease. It provides an up-to-date guidebook to navigate both conventional and highly specialized methods to detect and analyze the different signaling pathways of NF- B activation and contextualize them within organismal physiology and disease pathogenesis, using genetic and biochemical techniques and some of the most advanced computational and systems biology methods. Additionally, the volume includes several examples of approaches utilized by leading experts in the NF- B field to analyze and modulate NF- B signaling in specific physiological and disease contexts, along with some of the most promising approaches to pharmacologically target the NF- B pathway in human disease. Written for the highly successful 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, NF- B Transcription Factors: Methods and Protocols is an up-to-date guide intended for both basic and translational scientists who are working in the dynamic NF- B field.

Ever since the pioneering work of Darwin and Wallace, evolutionary biologists have attempted to understand the evolutionary dynamics of genetic systems. A range of theories on evolutionary ratesfrom static to gradual to punctuated to quantumhave been developed, primarily by comparing morphological changes over geological timescales as described in the fossil record. Recent studies, however, are beginning to change the way that we view evolutionary patterns and processes. New paleontological, experimental, molecular, and genomic investigations are providing a tremendous amount of novel data and fresh perspectives, offering valuable insights on the rates of evolutionary change, particularly in fast-evolving genetic systems. Rapidly Evolving Genes and Genetic Systems captures these recent exciting developments across a broad range of morphological, molecular, cellular, developmental, and genetic investigations in both natural and experimental populations over a diversity of life forms. The book provides a fascinating series of case studies that exemplify rapid evolution, and showcases the diversity of rapidly evolving genes and genetic systems, emphasizing the extremely important roles that they play in adaptation, speciation, and the generation and maintenance of a diversity of biological traits and properties. This exciting collection showcases the latest research of more than 50 eminent evolutionary biologists. It will be suitable for senior undergraduate students, graduate students, researchers, and for all those interested in the study of evolution.

Ever since the pioneering work of Darwin and Wallace, evolutionary biologists have attempted to understand the evolutionary dynamics of genetic systems. A range of theories on evolutionary ratesfrom static to gradual to punctuated to quantumhave been developed, primarily by comparing morphological changes over geological timescales as described in the fossil record. Recent studies, however, are beginning to change the way that we view evolutionary patterns and processes. New paleontological, experimental, molecular, and genomic investigations are providing a tremendous amount of novel data and fresh perspectives, offering valuable insights on the rates of evolutionary change, particularly in fast-evolving genetic systems. Rapidly Evolving Genes and Genetic Systems captures these recent exciting developments across a broad range of morphological, molecular, cellular, developmental, and genetic investigations in both natural and experimental populations over a diversity of life forms. The book provides a fascinating series of case studies that exemplify rapid evolution, and showcases the diversity of rapidly evolving genes and genetic systems, emphasizing the extremely important roles that they play in adaptation, speciation, and the generation and maintenance of a diversity of biological traits and properties. This exciting collection showcases the latest research of more than 50 eminent evolutionary biologists. It will be suitable for senior undergraduate students, graduate students, researchers, and for all those interested in the study of evolution.

The 'Adaptive Landscape' has been a central concept in population genetics and evolutionary biology since this powerful metaphor was first formulated by Sewall Wright in 1932. Eighty years later, it has become a central framework in evolutionary quantitative genetics, selection studies in natural populations, and in studies of ecological speciation and adaptive radiations. Recently, the simple concept of adaptive landscapes in two dimensions (genes or traits) has been criticized and several new and more sophisticated versions of the original adaptive landscape evolutionary model have been developed in response. No published volume has yet critically discussed the past, present state, and future prospect of the adaptive landscape in evolutionary biology. This volume brings together prominent historians of science, philosophers, ecologists, and evolutionary biologists, with the aim of discussing the state of the art of the Adaptive Landscape from several different perspectives.

This detailed book brings together a new set of protocols to arm cell biologists with techniques that are currently being used in a number of well-established laboratories around the world. The contents represent the great strides made in the field of cell-cell communications with respect to the identification and characterization of key components of the communication apparatus, assembly and maintenance of the communications structures, and concomitantly their roles in not only tissue formation and maintenance but also regeneration and repair. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective chapters, 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, Stem Cell Renewal and Cell-Cell Communication: Methods and Protocols, Second Edition serves as an ideal guide for experts and newcomers alike seeking to increase our understanding of the crucial biological and physiological roles of cell-cell communications in tissue function and organismal integrity.

Basics of proteins and proteomics techniques In-depth understanding of mass-spectrometry and quantitative proteomics An overview of interactomics and its application for translational research. Advancement in the field of proteomics and challenges in clinical applications.

The book presents comprehensive information on fundamental, and applied knowledge for developing varieties resistant individually as well as to all the major pathogens of crucifers, such as Albugo, Alternaria, Erysiphe, Hyaloperonospora, Plasmodiophora, Leptosphaeria, Sclerotinia, Turnip mosaic virus, Verticillium, and Xanthomonas through the use of latest biotechnological approaches including identification of R genes and their incorporation into agronomically superior varieties. The chapters include the information's viz., principles of host resistance, identification of R-genes sources, inheritance of disease resistance, host resistance signaling network system to multiple stresses. The book also covers transfer of disease resistance, and management of disease resistance. Standardized, reproducible techniques are also included for the researchers of cruciferous crops for developing resistant cultivars. The book deals with the gaps in understanding, knowledge of genomics, and offers suggestions for future research priorities in order to initiate the advance research on disease resistance. This book is immensely useful to the researchers especially Brassica breeders, teachers, extension specialists, students, industrialists, farmers, and all others who are interested to grow healthy, and profitable cruciferous crops all over the world.

The present book is an attempt to describe the most recent developments in the area of pericyte biology which is one of the emergent hot topics in the field of molecular and cellular biology today. Here, we present a selected collection of thirteen detailed chapters on what we know so far about pericytes in distinct organs in physiological and pathological conditions. Further, it provides an update on the most novel functions attributed to these cells and will introduce a newer generation of researchers and scientists to the importance of these cells, ranging from their discovery in different organs through current state-of-the-science. It will be invaluable for both advanced cell biology students as well as researchers in cell biology, stem cells and vascular research. This volume explores pericytes' physiologic roles in different tissues, ranging from the pancreas, lungs and liver through skeletal muscle, gut, retina and more. Together with its companion volumes Pericyte Biology in Disease and Pericyte Biology - Novel Concepts, Pericyte Biology in Different Organs presents a comprehensive update on the latest information and most novel functions attributed to pericytes. To those researchers newer to this area, it will be useful to have the background information on these cells' unique history. It will be invaluable for both advanced cell biology students as well as researchers in cell biology, stem cells and researchers or clinicians involved with specific organs.

This 2nd edition of the book on DNA methyltransferases has been comprehensively updated to reflect many novel research findings regarding the structure, function, and technology of these enzymes that have emerged over the past 6 years. Like the previous edition, this 2nd edition explains the biochemical properties of DNA methyltransferases, describing their structures, mechanisms and biological roles in bacteria, humans and plants. It also discusses the biological processes of reading DNA methylation and the mechanisms of DNA demethylation. This volume highlights the newest findings on DNA methyltransferase inhibitors and their use in cancer therapy as well as the latest epigenome editing systems based on these enzymes. Overall, this 2nd edition comprehensively summarizes the current state of research in the field of DNA methylation and DNA methyltransferase and is essential reading for early career and advanced researchers in this exciting field.

Statistical genomics is a rapidly developing field, with more and more people involved in this area. However, a lack of synthetic reference books and textbooks in statistical genomics has become a major hurdle on the development of the field. Although many books have been published recently in bioinformatics, most of them emphasize DNA sequence analysis under a deterministic approach.

"Principles of Statistical Genomics" synthesizes the state-of-the-art statistical methodologies (stochastic approaches) applied to genome study. It facilitates understanding of the statistical models and methods behind the major bioinformatics software packages, which will help researchers choose the optimal algorithm to analyze their data and better interpret the results of their analyses. Understanding existing statistical models and algorithms assists researchers to develop improved statistical methods to extract maximum information from their data.

Resourceful and easy to use, "Principles of Statistical Genomics" isa comprehensive reference for researchers and graduate students studying statistical genomics. "

This book focuses on the discoveries in M. truncatula genomic research which has been undertaken in the last two decades. Legumes are important for their economic values as food, feed, and fodder and also serve as the pillar of sustainable agriculture because of its biological nitrogen fixation capacity. Medicago truncatula was established as a model legume in the 1990s and has been well adopted as a model internationally since then. M. truncatula is an autogamous, diploid (2n = 16) species with a short generation time, and relatively small genome size (~375 Mbp). The M. truncatula genome was initially sequenced by the International Medicago Genome Annotation Group (IMGAG) in 2011 and has been well-annotated. M. truncatula research benefits from the availability of several genetic and genomic tools, such as gene expression atlas (MtGEA), insertion and neutron bombardment mutant populations, and a HapMap panel containing 384 sequenced inbred lines for genome-wide association studies. This book covers the current status and latest advancements of the M. truncatula genomics and transcriptomics resources along with a glimpse of newly developed tools that makes M. truncatula a front runner model in functional genomic studies.

This wide-ranging book introduces information as a key concept not only in physics, from quantum mechanics to thermodynamics, but also in the neighboring sciences and in the humanities. The central part analyzes dynamical processes as manifestations of information flows between microscopic and macroscopic scales and between systems and their environment. Quantum mechanics is interpreted as a reconstruction of mechanics based on fundamental limitations of information processing on the smallest scales. These become particularly manifest in quantum chaos and in quantum computing. Covering subjects such as causality, prediction, undecidability, chaos, and quantum randomness, the book also provides an information-theoretical view of predictability. More than 180 illustrations visualize the concepts and arguments. The book takes inspiration from the author's graduate-level topical lecture but is also well suited for undergraduate studies and is a valuable resource for researchers and professionals.

Biostimulants for crops from seed germination to plant development focuses on the effects and roles of natural biostimulants in every aspect of plant growth development to reduce the use of harmful chemical fertilizers and pesticides. Biostimulants are a group of substances of natural origin that offer a potential to reduce the dependency on harmful chemical fertilizers causing environmental degradation. While there is extensive literature on biostimulants, there remains a gap in understanding how natural biostimulants work and their practical application. This book fills that gap, presenting the ways in which biostimulants enhance seed vigor and plant productivity by looking into their mode of action, an area still being researched for deeper understanding. Exploring the roles of seed germination, pollen tube formation, pollen-pistil interaction, flower and fruit setting, to plant pigments, rhizospheric and soil microorganisms, the book also sheds light on the challenges and realistic opportunities for the use of natural biostimulants.

For more than two decades the concept of phenotypic plasticity has allowed researchers to go beyond the nature-nurture dichotomy to gain deeper insights into how organisms are shaped by the interaction of genetic and ecological factors. "Phenotypic Plasticity: Beyond Nature and Nurture" is the first work to synthesize the burgeoning area of plasticity studies, providing a conceptual overview as well as a technical treatment of its major components.

Phenotypic plasticity integrates the insights of ecological genetics, developmental biology, and evolutionary theory. Plasticity research asks foundational questions about how living organisms are capable of variation in their genetic makeup and in their responses to environmental factors. For instance, how do novel adaptive phenotypes originate? How do organisms detect and respond to stressful environments? What is the balance between genetic or natural constraints (such as gravity) and natural selection? The author begins by defining phenotypic plasticity and detailing its history, including important experiments and methods of statistical and graphical analysis. He then provides extended examples of the molecular basis of plasticity, the plasticity of development, the ecology of plastic responses, and the role of costs and constraints in the evolution of plasticity. A brief epilogue looks at how plasticity studies shed light on the nature/nurture debate in the popular media.

"Phenotypic Plasticity: Beyond Nature and Nurture" thoroughly reviews more than two decades of research, and thus will be of interest to both students and professionals in evolutionary biology, ecology, and genetics.

Beginning with the Escherichia coli ? protein, or bacterial DNA topoisomerase I, an ever-increasing number of enzymes have been identified that catalyze changes in the linkage of DNA strands. DNA topoisomerases are ubiquitous in nature and have been shown to play critical roles in most p- cesses involving DNA, including DNA replication, transcription, and rec- bination. These enzymes further constitute the cellular targets of a number of clinically important antibacterial and anticancer agents. Thus, further studies of DNA topology and DNA topoisomerases are critical to advance our und- standing of the basic biological processes required for cell cycle progression, cell division, genomic stability, and development. In addition, these studies will continue to provide critical insights into the cytotoxic action of drugs that target DNA topoisomerases. Such mechanistic studies have already played an important role in the development and clinical application of antimicrobial and chemotherapeutic agents. The two volumes of DNA Topoisomerase Protocols are designed to help new and established researchers investigate all aspects of DNA topology and the function of these enzymes. The chapters are written by prominent investigators in the field and provide detailed background information and st- by-step experimental protocols. The topics covered in Part I: DNA Topology and Enzymes, range from detailed methods to analyze various aspects of DNA structure, from linking number, knotting/unknotting, site-specific recombi- tion, and decatenation to the overexpression and purification of bacterial and eukaryotic DNA topoisomerases from a variety of cell systems and tissues.

This book presents a comprehensive discussion on the novel concepts in stem cell heterogeneity, from pluripotent stem cells to human mesenchymal stem cells, adult and cancer stem cells of the thyroid, sarcoma, and more. Thus, Stem Cell Heterogeneity - Novel Concepts starts from a timely update on the current information on stem cells heterogeneity in various tissues and discusses new concepts and future directions. It also provides a solid foundation of the history of stem cells from specific tissues and the current applications of this knowledge in regenerative medicine. When taken as a whole, alongside its companion volumes Stem Cells Heterogeneity in Different Organs, and Stem Cells Heterogeneity in Cancer, these three books present a comprehensive reference on stem cell heterogeneity in various tissues and current and future applications for regenerative medicine. It is essential reading for advanced cell biology students as well as researchers in stem cells and clinicians.

Pillars of Evolution provides a fresh and provocative perspective on adaptive evolution. Readers new to the study of evolution will find a refreshing new insight that establishes evolutionary biology as a rigorous and predictive science, whilst practicing biologists will discover a provocative book that challenges traditional approaches.
The book begins by leading readers through the mechanics of heredity, reproduction, movement, survival, and development. With that framework in place, it then explores the numerous ways that traits emerge from the interactions between genetics, development, and the environment. The key message is that adaptive changes in traits (and their underlying allelic frequencies) evolve through the traits' functions and their connection with fitness. The complex mappings from genes-to-traits-to-fitness are characterized in the structure of evolution. A single "structure matrix" describes why individuals vary in the values of adaptive traits, their ability to perform the function of those traits, and in the fitness they accrue. Fitness depends on how organisms interact with and perceive their environment in time and space. These relationships are made explicit in spatial, temporal, and organizational scale that also sets the stage for the crucially important role that ecology always plays in evolution. The ecological hallmarks of density- and frequency-dependent interactions allow the authors to explore new and exciting insights into evolution's dynamics. The theories and principles are then brought together in a final synthesis on adaptation.
The book's unique approach unites genetic, development, and environmental influences into a single comprehensive treatment of the eco-evolutionary process.

The history of life is a nearly four billion year old story of transformative change. This change ranges from dramatic macroscopic innovations such as the evolution of wings or eyes, to a myriad of molecular changes that form the basis of macroscopic innovations. We are familiar with many examples of innovations (qualitatively new phenotypes that can provide a critical benefit) but have no systematic understanding of the principles that allow organisms to innovate. This book proposes several such principles as the basis of a theory of innovation, integrating recent knowledge about complex molecular phenotypes with more traditional Darwinian thinking. Central to the book are genotype networks: vast sets of connected genotypes that exist in metabolism and regulatory circuitry, as well as in protein and RNA molecules. The theory can successfully unify innovations that occur at different levels of organization. It captures known features of biological innovation, including the fact that many innovations occur multiple times independently, and that they combine existing parts of a system to new purposes. It also argues that environmental change is important to create biological systems that are both complex and robust, and shows how such robustness can facilitate innovation. Beyond that, the theory can reconcile neutralism and selectionism, as well as explain the role of phenotypic plasticity, gene duplication, recombination, and cryptic variation in innovation. Finally, its principles can be applied to technological innovation, and thus open to human engineering endeavours the powerful principles that have allowed life's spectacular success.