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This current edition explores new tests for genotoxicity testing, along with other well-known techniques. This will further help in our understanding of the genotoxic effects of chemicals. The book has different sections dealing with various assays for gene mutation, chromosomal abnormalities, primary DNA damage, etc. It also delves into plant models, animals and their alternates, as well as in silico approaches for genetic toxicology. 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, tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Genotoxicity Assessment: Methods and Protocols, Second Edition serves as a highly useful and ready resource for research students and scientists working in regulatory toxicology as well as biomedical, biochemical, and pharmaceutical sciences.

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The book focuses on the regulation of protein function and biological activity by the post-translational modification known as acetylation. Chapters detail a wide range of topics in protein acetylation dealing with pathways relevant to cellular homeostasis, gene expression, and DNA metabolism. 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, Protein Acetylation: Methods and Protocols aims to be an extremely valuable resource to novices and experts alike whose research takes them in the direction of characterizing regulatory switches in protein acetylation events, and how they are manifested in significant ways in diverse mechanisms biological systems.

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The natural, biological, medical, and related sciences would not be what they are today without the microscope. After the introduction of the optical microscope, a second breakthrough in morphostructural surface analysis occurred in the 1940s with the development of the scanning electron microscope (SEM), which, instead of light (i. e. , photons) and glass lenses, uses electrons and electromagnetic lenses (magnetic coils). Optical and scanning (or transmission) electron microscopes are called “far-field microscopes” because of the long distance between the sample and the point at which the image is obtained in comparison with the wavelengths of the photons or electrons involved. In this case, the image is a diffraction pattern and its resolution is wavelength limited. In 1986, a completely new type of microscopy was proposed, which, without the use of lenses, photons, or electrons, directly explores the sample surface by means of mechanical scanning, thus opening up unexpected possibilities for the morphostructural and mechanical analysis of biological specimens. These new scanning probe microscopes are based on the concept of near-field microscopy, which overcomes the problem of the limited diffraction-related resolution inherent in conventional microscopes. Located in the immediate vicinity of the sample itself (usually within a few nanometers), the probe records the intensity, rather than the interference signal, thus significantly improving resolution. Since the most we- known microscopes of this type operate using atomic forces, they are frequently referred to as atomic force microscopes (AFMs).

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This volume provides methods to study bone morphogenetic proteins (BMPs) and BMP modulators in cell culture, in both invertebrate and vertebrate animal models, and for therapeutic applications. Chapters guide the reader in the use of primary and immortalized cell lines, methodologies that exploit transcription and receptor mechanisms, newly developed animal models using nematodes, flies, amphioxus, frogs, zebrafish, and genetically engineered mice, and pre-clinical approaches to understanding BMP function in bone regeneration, heterotopic ossification, and cancers. 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 cutting-edge, Bone Morphogenetic Proteins: Methods and Protocols aims to ensure successful results in the further study of this vital field.

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This detailed volume addresses the challenge of how to instruct stem/early progenitor cells to progress through appropriate steps to generate functional 3-dimensional organs, one of the outstanding issues in regenerative medicine. The field of organoids is geared towards defining and demonstrating the in vitro conditions that achieve this goal. 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. Comprehensive and cutting-edge, Organoids: Stem Cells, Structure, and Function serves as an aid to researchers working in this vital area of research.

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This volume presents standard approaches and the most recent technical advances used to study innate immune activation. Chapters detail the assessment of macrophage activation, measuring innate immune responses to bacterial viability, quantification of secreted proteins, reporter systems, protocols examining specific innate immune activation by TLRs, RLRs, cGAS, and inflammasomes. 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 practical, Innate Immune Activation: Methods and Protocols aims to be a useful and informative reference tool for further study into this vital field.

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With the completion of a finished rice genome sequence, increasing efforts have focused on functional characterization of rice genes, elucidation of the underlying mechanisms involved in major agronomic traits (e.g., high yield, grain quality, abiotic stress tolerance, and disease resistance), and the subsequent translation of genomic knowledge into agricultural productivity via molecular breeding and improved cultural practice. To meet increasing interest in this field, Rice Protocols has been compiled to provide a series of core techniques and approaches commonly used in studying rice molecular biology and functional genomics. These approaches include genetic and molecular techniques such as artificial hybridization, fluorescence in situ hybridization, generation and characterization of chemical and T-DNA insertional mutants, quantitative trait loci (QTLs) analysis and map-based cloning, site-specific transgene integration, and artificial microRNA-mediated gene silencing, along with a variety of “omics” techniques. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and easy to use, Rice Protocols will prove useful for both beginners and experienced researchers whether they are molecular biologists who want to study rice plants or rice researchers who are interested in learning molecular techniques.

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A critical review of the classic, as wells as most recent-and quite seminal-findings concerning the phenotypic and molecular characteristics of both fetal and neonatal B and T cells, the cells that mediate antibody and cellular immune responses in newborns and infants. Dr. Bona shows how the antibody response of neonates is modulated by maternal antibodies and how, in certain cases, this can cause transient or life-threatening neonatal autoimmune disease. He also describes the characteristics of neonatal tolerance induced by foreign allo- and self-antigens, which are the basis for understanding impaired infant immune response and which provide a rationale for the development of efficient neonatal vaccines. By making clear the characteristics and differences between the immune system and the immune responses of both newborns and infants, compared to those of adults, Dr. Bona offers insights and challenging hypotheses that promise to help overcome the poor responses of neonates to various antigens.

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In this greatly enlarged and thoroughly updated edition of his much praised Cardiac Markers, Alan Wu and his contributors focus on the use of markers in the practice of cardiology and-for the first time-on the use of natriuretic peptides for congestive heart failure. Here, leading international authorities in clinical chemistry and laboratory medicine, cardiology, emergency medicine, and the in vitro diagnostics industry describe the state-of-the-art uses of cardiac markers when treating coronary artery disease, and discuss in detail how they may be optimally used in a clinical setting. Comprehensive and cutting-edge, Cardiac Markers, Second Edition offers physicians a complete guide to the use of cardiac markers in clinical practice and clinical laboratorians a close-up view of the new markers now becoming standard.

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This detailed second edition volume expands upon the prior edition by addressing newly emerged technologies as well as improved existing protocols in nucleic acid delivery via nanotechnology. The book addresses topics ranging from chemical synthesis of macromolecules and bioconjugates, novel and established nanoformulations, characterization of these nanoformulations for biophysical, biological and toxicological aspects, and also protocols dealing with application and imaging of such carrier systems in vivo. 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 up-to-date, Nanotechnology for Nucleic Acid Delivery: Methods and Protocols, Second Edition serves as an ideal guide to researchers seeking to use this vital area of study in their own lab work or to progress knowledge within the field itself.

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This book aims to provide a guide to researchers on how to work with Transient Receptor Potential (TRP) channels, describing current methodologies and protocols, and highlighting the challenges ahead that will require the development of new technologies. Articles cover a wide range of techniques, from structure to function, concluding with how to best to evaluate some of the in vivo function of TRP channels, particularly in regard to their involvement in hyperalgesia and allodynia. Also, the collection describes important and inspiring available tools that are currently being used to investigate the activity of these channels in vitro and in vivo in terms of their underlying protein structure, segueing into dealing with the generation of pharmacological tools to explore their in vivo activity and their involvement in cellular signaling pathways, which may be developed into drugs to treat diseases caused by channel dysfunction or hyperactivity. Written for the highly successful Methods in Molecular Biology series, chapters include the necessary detail and implementation advice to ensure successful results in the lab. Authoritative and practical, TRP Channels: Methods and Protocols is a useful set of methodologies for scientists seeking to better understand this vital family of channels.

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This volume covers several aspects of rational drug design, such as synthesis of novel bioactive drugs; development and application of new methodologies; computational methods valuable for the establishment of new approaches in drug discovery; and the effects of physical-chemical and ADMET properties of the designed potential drugs. Chapters guide readers through amyloid deposits, Saturation Transfer Difference (STD) NMR, methods on bioguided design, the importance of lipophilicity in drug design, ADMET, FRET, structural biology, and homology modeling. 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, Rational Drug Design: Methods and Protocols aims to ensure successful results in the further study of this vital field.

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This volume provides readers with a better understanding of organogenesis in developmental biology and next-generation organ regenerative therapy. This book focuses on recent studies of organ regeneration from stem cells using in vitro 3D cell culture and manipulation. The chapters cover topics such as generation of a 3D retinal tissue formation; functional salivary gland regeneration; liver regeneration using cultures liver bud; and in vivo model of small intestine. 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.Comprehensive and thorough, Organ Regeneration: 3D Stem Cell Culture and Manipulation is a valuable resource for scientists and researchers who are interested in this field.

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This volume presents an overview of recent advances, innovative applications, and future prospects of in vitro embryogenesis in higher plants. The book's chapters are divided into five parts: Part I contains reviews on general topics (microspore; zygotic and somatic embryogenesis; in vitro and in vivo asexual embryogenesis; advances on the genetic, physiological, and proteomic knowledge of somatic embryo formation; role of apoptosis and mitochondria in somatic embryogenesis; and innovation in the use of bioreactors). The remaining four parts discuss step-wise protocols on somatic embryogenesis in selected horticultural plants (Part II); forest trees (Part III); gametic embryogenesis (Part IV); and pivotal topics, such as the detection of epigenetics modifications during microspore embryogenesis, the in vitro embryogenesis and plant regenerations from isolated zygotes, the synthetics seed production, the induction and maturation of somatic embryos, and the cryostorage of embryogenic cultures (Part V). 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, In Vitro Embryogenesis in Higher Plants, is a useful source of information and ideas for plant tissue culturists, cell biologists, embryologists, horticulturists, and operators of commercial nurseries. This book will introduce the fascinating work in in vitro embryogenesis in higher plants to students and young scientists.

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Filamentous fungi have long been known for their ability to produce an enormous range of unusual chemical compounds known as secondary metabolites, many of which have potentially useful antibiotic or pharmacological properties. Recent focus on fungal genomics coupled with advances in detection and molecular manipulation techniques has galvanized a revitalization of this field. Fungal Secondary Metabolism: Methods and Protocols is aimed at providing the key methodologies currently in use and necessary for accessing and exploiting the natural product information provided by the genomes of this large and varied kingdom. Written by active researchers in the field, the chapters deal with all the steps necessary, from optimization of fungal culture conditions for metabolite production, through rapid genome sequencing and bioinformatics, and genetic manipulations for functional analysis, to detection and testing of metabolites. In addition, chapters on basic science address approaches to the genetic regulation, protein biochemistry, and cellular localization of the biosynthetic pathways. 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 hands-on, Fungal Secondary Metabolism: Methods and Protocols encourages new investigators to enter the field and expands upon the expertise and range of skills of those already researching fungal natural products.

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The second edition of this volume focuses on applied bioinformatics with specific applications to crops and model plants. Plant Bioinformatics: Methods and Protocols is aimed at plant biologists who have an interest in, or requirement for, accessing and manipulating huge amounts of data being generated by high throughput technologies. This book would also be of interest to bioinformaticians and computer scientists who would benefit from an introduction to the different tools and systems available for plant research. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and software, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and thorough, Plant Bioinformatics: Methods and Protocols helps researchers with the increasing volume and diversity of data from different plants and also the integration of multiple diverse forms of data.

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Engineering Multicellular Systems: Methods and Protocols, focuses on laboratory procedures used in recent efforts for constructing synthetic multicellular systems and their applications. In particular, constructing multicellular systems to form various microbial ecosystems has been extensively explored to examine evolution and interactions of microbial ecosystems, while co-cultures have emerged as an efficient tool to produce some complex chemical molecules. 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.Engineering Multicellular Systems: Methods and Protocols provide a comprehensive laboratory protocol reference for constructing multicellular systems for various applications.

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The application of molecular techniques to gastroenterology continues to yield important advances in the development of drugs to treat gastrointestinal disorders. Important new drugs have emerged through the collaborative and complementary efforts of basic scientists, clinicians, and clinical researchers in academia and the pharmaceutical industry. The challenge has been exciting, with a few surprises along the way. Consider peptic ulcer disease as an example. The discovery of H receptors and the availability of potent and 2 selective H-receptor antagonists signaled the beginning of a new era 2 in the treatment of gastric hypersecretory states and peptic ulcers. Introduction of proton pump inhibitors offered another therapeutic option. Though H-receptor antagonists and proton pump inhibitors 2 are important and useful drugs, the discovery of the link between H. pylori infection and peptic ulcer disease has led to even more effective pharmacotherapeutic regimens. Our intent in Drug Development: Molecular Targets for GI Diseases is to bring together hands-on experts to review promising areas of gastrointestinal pharmacology. The contemporary topics covered, from a mechanistic viewpoint, are relevant to gastrointestinal inflammation and motility disorders. Authoritative opinions are offered on both future research directions and potential applications for new therapies.

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Physicians, nurses, and safety experts comprehensively review sedation and analgesia to provide a completely new reference guide to safe sedation practices consistent with existing guidelines. Starting with an integrated review of the basic physiology and neurobiology of the sedated state, the authors proceed through clinical guidelines and practices, and conclude with an examination of quality-outcome measures and processes. They also review current mandates for safe sedation practices and address the key clinical issues of pharmacology, monitoring, and recovery. Special tables and figures throughout the book summarize protocols, regulatory requirements, recommended dosages, monitoring requirements, and quality assurance tools.

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