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This volume covers comprehensive methods on ways to assess structural and ultrastructural changes in the mitochondria, cytoskeleton, and microglia using state-of-the-art microscopy techniques including super-resolution imaging, electron microscopy, and ultra-high field MRI. The chapters in this book cover topics such as analysis of neurodegeneration in the post-mortem characterization of preclinical animal models, in vivo modeling in cell death in different model systems and brain organoids, single cell clonal analysis using Mosaic Analysis with Double Markers in genetic mouse models, and genome and proteomic methods for analysis of mRNA dynamics and quantitation of targeted peptides. 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.Cutting-edge and thorough, Neuronal Cell Death: Methods and Protocols is a valuable resource for any scientist and researcher interested in learning more about this developing field.
This detailed volume explores protocols for the production of membrane proteins in a panel of heterologous organisms for structural studies. Beginning with techniques using E. coli as a host for the overproduction and purification of membrane proteins, the book continues with chapters covering mammalian membrane protein production in yeast, insect cells, mammalian cells, as well as using virus like particles and acellular systems. Additionally, new detergents and alternatives to detergents allowing membrane protein purification for structural analyses are described. The book closes with a chapter exploring the use of microscale thermophoresis (MST) to evaluate the binding activity of heterologously expressed proteins directly in crude membrane extracts. 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, Heterologous Expression of Membrane Proteins: Methods and Protocols, Third Edition serves as an ideal guide for scientists aiming to produce and purify functional recombinant membrane proteins for structural studies.
This volume details comprehensive protocols and methodologies to assess mitochondrial bioenergetics and dynamics in different tissues and cells involving health and pathological states. Chapters guide readers through methods for assessment of the energy metabolism including Oxygen Consumption Rate (OCR), mitochondrial membrane potential, and measuring mitochondrial Ca2+ handling, and ROS emission. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting, and systematic reproducible protocols. Authoritative and cutting-edge, Mitochondria: Methods and Protocols aims to be a foundation for future studies and to be a source of inspiration for new investigations in the field.
This volume details a collection of laboratory protocols to study asthma in mice and humans. Chapters cover animal models of asthma, methods to measure asthma-related molecules, protocols to detect, isolate, culture and stimulate cells that contribute to asthma in mice and humans, methods to deplete these cells in vivo and study responses of intact airway tissues ex vivo. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Asthma: Methods and Protocols aims to be a foundation for future studies and to be a source of inspiration for new investigations in the field.
This volume provides detailed protocols for the identification and understanding of vasculogenic mimicry process in vitro and in vivo, in addition to protocols for microscopy and histology. Chapters guide readers through different materials, commercial and homemade scaffolds, Matrigel, cancer spheroids, 3D tissue constructs, vasculogenic processes, and mathematical model building. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Vasculogenic Mimicry: Methods and Protocols aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
This detailed volume provides a comprehensive collection of methods and protocols in food allergy and food allergens studies. The selected protocols explore the study of food allergens, from recombinant production, purification procedures, IgE and T cell epitopes characterization, to allergen structure description, cellular responses, and tolerance induction, through a variety of techniques and animal models. 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 and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Food Allergens: Methods and Protocols serves as an ideal reference for scientists at all stages involved in the study of food allergy and allergenic components.
This volume explores the application of high-performance computing (HPC) technologies to computational drug discovery (CDD) and biomedicine. The first section collects CDD approaches that, together with HPC, can revolutionize and automate drug discovery process, such as knowledge graphs, natural language processing (NLP), Bayesian optimization, automated virtual screening platforms, alchemical free energy workflows, fragment-molecular orbitals (FMO), HPC-adapted molecular dynamic simulation (MD-HPC), and the potential of cloud computing for drug discovery. The second section delves into computational algorithms and workflows for biomedicine, featuring an HPC framework to assess drug-induced arrhythmic risk, digital patient applications relevant to the clinic, virtual human simulations, cellular and whole-body blood flow modeling for stroke treatments, prediction of the femoral bone strength from CT data, and many more subjects. Written for the highly successful Methods inMolecular Biology series, chapters include introductions to their respective topics, lists of the necessary software and tools, step-by-step and readily reproducible modeling protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, High Performance Computing for Drug Discovery and Biomedicine allows a diverse audience, including computer scientists, computational and medicinal chemists, biologists, clinicians, pharmacologists and drug designers, to navigate the complex landscape of what is currently possible and to understand the challenges and future directions of HPC-based technologies.
This second edition details new and updated protocols that cover techniques used to study secretion systems. Chapters focus on identifying and localizing the different subunits, defining interactions within subunits, monitoring conformational changes, purifying and imaging of large complexes, defining the assembly pathway by fluorescence microscopy and the role of energy during assembly and/or secretion, identifying secreted effectors as well as using reporters to follow effector transport. 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, Bacterial Secretion Systems: Methods and Protocols- Second Edition aims to be a useful andpractical guide to new researchers and experts looking to expand their knowledge.
This second edition provides new and updated methods and techniques for identification of drug target, binding sites prediction, high- throughput virtual screening, lead discovery and optimization, conformational sampling, prediction of pharmacokinetic properties using computer-based methodologies. Chapters also focus on the application of the latest artificial intelligence technologies for computer aided drug discovery. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Computational Drug Discovery and Design, Second Edition aims to effectively utilize computational methodologies in discovery and design of novel drugs.
This detailed volume presents a series of protocols that are representative of recent developments and improvements in induced pluripotent stem cells (iPS cells) and corresponding human disease models. Reflecting the latest technology for generating induced pluripotent stem cells (iPS cells) and their initial characterization, the book explores techniques invaluable both for studies of disease-specific cell types and for their potential applications in regenerative medicine. Written for the highly successful Methods in Molecular Biology series, chapters include introduction to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, as well as tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Induced Pluripotent Stem Cells and Human Disease: Methods and Protocols serves as a vital guide that is valuable for not only experts but also novices in the stem cell field.
This volume covers the latest developments in different areas of plant pathology. The chapters in this volume are organized into seven parts. Part One provides traditional methods for isolation and identification of invasive pathogens and root disease. Part Two looks at new and rapid DNA extraction protocols from different samples, and Part Three focuses on molecular detection protocols for identifying and quantifying plant pathogens, including fungal and bacterial invasive species. Part Four describes the application of metabarcoding in plant pathology, and Part Five talks about plant pathogen interactions. Part Six concentrates on population genomics of plant pathogens, and Part Seven covers biocontrol on plant pathogens. 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.Cutting-edge and thorough, Plant Pathology: Methods and Protocols is a valuable resource for researchers in the plant pathology community, and discusses various approaches for the detection, identification, and control of plant diseases.
This second edition volume expands on the previous edition with new and updated chapters on the latest developments in the study of yeast within the biotechnology field. The chapters in this book cover topics such as transformation protocols for genetic engineering of Saccaromyces cerevisiae and Komagataella spp.; an overview of selection markers, promoters, and strains used for metabolic engineering of S. cerevisiae, P. pastoris, and Z. bailii; the use of yeast in CRISPR/Cas9 technology; tools to study metabolic pathway in Yarrowia lypolitica; and a discussion on the "e;universal expression system"e; that is applied in a broad spectrum of fungal species. 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.Cutting-edge and authoritative, Yeast Metabolic Engineering: Methods and Protocols, Second Edition is a valuable resource for researchers and scientists interested in learning more about this important and developing field.
This detailed book presents recent methodologies for the task of inspecting the genomic world of plants, extracting valuable information, and presenting it in a readable way. With a focus on bioinformatics tools, the volume explores phylogenetics and evolution, Omics analysis, as well as experimental procedures for trait characterization. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of vital expert implementation advice that will lead to successful results. Authoritative and practical, Plant Comparative Genomics serves as an ideal resource for researchers looking to implement comparative tools in order to explore their genomic data for their daily scientific work.
This detailed volume explores the application of multiplex biomarker methods in the critical area of COVID-19 research through state-of-the-art technologies in the fields of genomics, proteomics, transcriptomics, metabolomics, and imaging. The book features a series of protocols from labs across the globe employing multiplex molecular approaches, which can be applied to accelerate progress in the research of SARS-CoV-2 and other infectious illnesses. 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 essential, Multiplex Biomarker Techniques: Methods and Applications for COVID-19 Disease Diagnosis and Risk Stratification serves as a vital resource for researchers in the areas of virology, metabolic diseases, respiratory disorders, as well as to clinical scientists, physicians, pharmacologists, and the healthcare services.
This detailed volume explores newly-developed methods in PIWI-interacting RNAs (piRNAs) research, methods currently applied to other ncRNAs involved in nuclear regulation which can be used to study piRNAs, and piRNA methods applied in non-classical organisms. It also includes several bioinformatic and biophysical methods related to piRNA studies, consistent with the increasing importance of high-throughput sequencing and computational methods. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, piRNA: Methods and Protocols serves as an ideal guide for researchers seeking to elucidate the numerous mysteries of this area of multicellular biology.
This detailed volume presents a series of methods exploring membrane trafficking research, ranging from genetics and high-resolution imaging to in vitro biochemical and biophysical assays. Covering virtually all the major trafficking branches, the book delves into the exocytic pathway, which focuses on cargo transport from the ER to the Golgi, through the Golgi cisternae, and to the plasma membrane and the extracellular space; the endocytic pathway, which includes cargo endocytosis, endosomal recycling, and lysosomal degradation; as well as emerging topics beyond the conventional exocytic and endocytic pathways. 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, Membrane Trafficking: Methods and Protocols provides techniques with broad applications as an ideal guide for junior researchers new to membrane trafficking as well as established membrane biologists seeking to expand their research programs.
This detailed volume examines the complex study of the assessment of in situ bioavailability and toxicity of organic chemicals in aquatic systems with a toolbox of reliable techniques. Beginning with a section on approaches for chemical analytical and bioanalytical techniques in bioavailability research, the book continues with methods to monitor effects in situ and conduct bioassays to assess the effects of complex environmental samples. It concludes with descriptions of various computational models. Written for the Methods in Pharmacology and Toxicology series, chapters feature the kind of expert implementation advice that leads to greater success in the field. Authoritative and versatile, In Situ Bioavailability and Toxicity of Organic Chemicals in Aquatic Systems serves as an ideal guide to aid in tackling the challenge of analyzing and understanding chemical pollution in aquatic systems.
This volume provides updated protocols for chemical protein synthesis. Chapters guide readers through development methods, strategies, and applications of protein chemical synthesis. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols. Authoritative and cutting-edge, Chemical Protein Synthesis aims to be a useful and practical guide to new researchers and experts looking to expand their knowledge.
This volume presents a collection of protocols to study effector-triggered immunity (ETI) in both plants and animals from eminent groups in the field. The chapters in this book cover topics such as genetic manipulation of plant and animal pathogens, host cells, and the analysis of key host responses; and techniques used for the analysis of inflammasome activation, cell death pathways, and mitochondria damage in response to pathogens. All of these topics cover a broad spectrum of immunological, biochemical, cell biological, and structural biology approaches to examine ETI. 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.Cutting-edge and practical, Effector-Triggered Immunity: Methods and Protocols is a valuable resource for both expert and novice researchers who are interested in learning more about the important and developing field of ETI.
Polymer science is a technology-driven science. More often than not, technological breakthroughs opened the gates to rapid fundamental and theoretical advances, dramatically broadening the understanding of experimental observations, and expanding the science itself. Some of the breakthroughs involved the creation of new materials. Among these one may enumerate the vulcanization of natural rubber, the derivatization of cellulose, the giant advances right before and during World War II in the preparation and characterization of synthetic elastomers and semi crystalline polymers such as polyesters and polyamides, the subsequent creation of aromatic high-temperature resistant amorphous and semi-crystal line polymers, and the more recent development of liquid-crystalline polymers mostly with n~in-chain mesogenicity. other breakthroughs involve the development of powerful characterization techniques. Among the recent ones, the photon correlation spectroscopy owes its success to the advent of laser technology, small angle neutron scattering evolved from n~clear reactors technology, and modern solid-state nuclear magnetic resonance spectroscopy exists because of advances in superconductivity. The growing need for high modulus, high-temperature resistant polymers is opening at present a new technology, that of more or less rigid networks. The use of such networks is rapidly growing in applications where they are used as such or where they serve as matrices for fibers or other load bearing elements. The rigid networks are largely aromatic. Many of them are prepared from multifunctional wholly or almost-wholly aromatic kernels, while others contain large amount of stiff difunctional residus leading to the presence of many main-chain "liquid-crystalline" segments in the "infinite" network.
This book represents the proceedings of the First International Conference on Frontiers of Polymer Research held in New Delhi, India during January 20-25, 1991. Polymers have usually been perceived as substances to be used in insulations, coatings, fabrics, and structural materials. Defying this classical view, polymers are emerging as a new class of materials with potential applications in many new technologies. They also offer challenging opportunities for fundamental research. Recognizing a tremendous growth in world wide interest in polymer research and technology, a truly global "1st International Conference on Frontiers of Polymer Research" was organized by P. N. Prasad (SUNY at Buffalo), F. E. Karasz (University of Massachusetts) and J. K. Nigam (Shriram Institute for Industrial Research, India). The 225 participants represented 25 countries and a wide variety of academic, industrial and government groups. The conference was inaugurated by the Prime Minister of India, Mr. Chandra Shekhar and had a high level media coverage. The focus of the conference was on three frontier areas of polymer research: (i) Polymers for photonics, where nonlinear optical properties of polymers show great promise, (ii) Polymers for electronics, where new conduction mechanisms and photophysics have generated considerable enthusiasm and (iii) High performance polymers as new advanced polymers have exhibited exceptionally high mechanical strength coupled with light weight.
Several hundred nuclear scientists from more than a dozen countries met in Miami Beach, Florida, in September 1989 for a Symposium on Exotic Nuclear Spectroscopy, sponsored by the Division of Nuclear Chemistry and Technology of the Amencan Chemical Society. Braving five days of beckoning beaches, they presented, listened to, and discussed a series spectroscopy-both experimental and of invited papers covering the renaissance of nuclear theoretical-that has occurred during the past decade and which promises to continue well into the decade to come. This book contains the Proceedings of that Symposium on Exotic Nuclear Spectroscopy. But it is much more: During the ensuing six months, most of the pa{>ers were rewritten, polished, and/or expanded; so the resulting book is a much more fimshed, comprehensive, and up-to-date product than a general proceedings. Chapter se~uences follow the original symposium program, which, with a few exceptions, juxtaposes like topics. Each chapter can stand more or less on its own-although, as in any complex, interrelated scientific field, there are many cross-references among chapters. The manuscripts were edited and then uniformly typed in the Department of Chemistry at Michigan State University. I tried to keep the editing to a minimum so as to preserve the flavor of individual styles. A great debt of gratitude goes to those who helped make this volume possible, especially to Vada O'Donnell, who bore the largest burden of typing (and clarifying) the manuscripts.
This volume details protocols on mass spectrometry and associated techniques. Chapters guide readers through micro- and macronutrients analysis, mass spectrometry-related methodologies, direct insertion, matrix-assisted laser desorption ionization (MALDI), gas chromatography (uni- and bi-dimensional), liquid chromatography, plasma mass spectrometry (ICP-MS), and analyses in food samples. Authoritative and cutting-edge, Mass Spectrometry for Food Analysis aims to provide comprehensive and updated state-of-art methodologies and models for food analysis.
This volume provides practical guidance on a variety of techniques and steps to ensure successful variant calling. Chapters detail methods for variant calling from single-nucleotide variants to structural variants, variant calling in specialized data types such as RNA-seq and UMI-tagged sequencing, alignment-free genotyping and SNP calling, variant detection in single-cell DNA sequencing data, variant annotation, and preanalytical quality control to ensure successful variant calling. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists step-by-step protocol to execute the algorithms, describes the input and output data, and includes tips on troubleshooting and known pitfalls. Authoritative and cutting-edge, Variant Calling: Methods and Protocols aims to be a foundation for future studies and to be a source of inspiration for new investigations in the field.
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