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This detailed book focuses on cutting-edge methods for Parkinson¿s disease (PD) research, such as the analysis of the prion-like properties of ¿-synuclein, mitochondrial functions related to the PINK1-Parkin pathway/CHCHD2, the endolysosome pathway related to LRRK2, VPS35, and ATP13A2 using cultured cells (including patient iPS cells), deep brain stimulation therapy, classic mitochondrial toxins related to PD, and genetic associations and screenings using mammalian and invertebrate genetic models of PD. The collection intends to serve as an introductory protocol book for basic research on PD pathogenesis. 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, Experimental Models of Parkinson's Disease is an ideal guide for researchers continuing to work toward making PD risk predictable and surmountable.
This volume of "Encyclopedia of Sustainability Science and Technology, Second Edition," covers the electrification of vehicles, which is key to a sustainable future of transportation in both light-duty and heavy-duty vehicle sectors to address global concerns of climate change, air pollutant emissions, energy efficiency and energy security. Vehicle electrification includes several existing and emerging technologies and powertrain architectures such as conventional hybrid electric vehicles (HEVs), plug-in hybrids with various electric driving range, short- and long-range battery electric vehicles, as well as hydrogen fuel cell electric vehicles (FCEVs). Electrification will be key to connected autonomous vehicles, which are perceived to improve mobility, increase safety, reduce energy consumption and infrastructure costs, improve productivity, decrease traffic congestion and increase customer satisfaction. While electrification of vehicle technologies is relatively mature, technology improvement and economies of scale are needed to compete against incumbent technologies and to realize their benefits in the marketplace. Significant infrastructure development is needed in the case of hydrogen fuel cell vehicles and to a lesser extent for plug-in electric vehicles.Vehicle efficiency improvement is sought through a combination of several approaches, including weight reduction, engine downsizing, increased engine compression ratio with high octane fuels, and the use of compression ignition engines with low octane fuels. Liquid hydrocarbon fuels are needed in applications where high storage energy density is required such as long-haul class-8 combination heavy-duty trucks. Shared mobility is another emerging concept that enables access to transportation services on an as-needed basis. This approach can enhance accessibility to transportation, decrease number of vehicles on the road, reduce energy use and impact on the environment, reduce cost of transportation and the need for parking, and reduce transportation time between origin and destination. In all, the reader will receive a comprehensive introduction to electric vehicles and technology trends, including energy storage, in light-, medium-, and heavy-duty sectors, as well as the infrastructure development that will be required to realize these benefits for society.
This detailed volume presents a variety of animal models that are commonly used to study sepsis and some key procedures to measure specific disease outcomes. The chapters describe well-established surgical and nonsurgical rodent models of sepsis, presented by experts in the field. In addition, the book includes protocols for burn injury and sepsis, modeling the mouse intensive care unit (MICU), and the development of humanized mice, which may be useful tools to increase the translational potential of rodent sepsis research. There is a chapter discussing the use of biomarkers for sepsis diagnosis and prognosis in humans and mice, as well as chapters describing specific methods for biomarker measurement, and the use of analgesics and humane endpoints in rodent sepsis research. Finally, agent-based computational modeling is presented as a valuable complementary approach to study sepsis. 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, Sepsis: Methods and Protocols serves as an ideal guide for researchers working toward improving the identification and treatment of septic patients.
This second edition provides new and updated chapters detailing recent advances in MYC research and current techniques. Chapters guide readers through protocols on how to express and purify MYC protein, X-ray crystallography, NMR, techniques to study how MYC is modified, apoptosis, senescence, proliferation, metabolic changes, translation, tumorigenesis,reprogramming, and clinical application of MYC studies.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, The Myc Gene: Methods and Protocols, Second Edition aims to ensure successful results in the further study of this vital field.
This second edition volume expands on the previous edition with updated chapters in model systems and new crops. The book contains protocols for plastid engineering in leaves, tissue culture cells, and the shoot apex of plants, as well as for marker excision from the plastid genome and engineering Rubisco, the key enzyme of photosynthesis. 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, Chloroplast Biotechnology: Methods and Protocols, Second Edition is a valuable resource for researchers who wish to enter the field, and for practitioners looking for insights of applications in agriculture, industrial biotechnology and healthcare.
This volume provides an overview of the current successes as well as pitfalls and caveats that are hindering the design of membrane proteins. Divided into six parts, chapters detail membrane transporter, FoldX force field, protein stability, G-Protein Coupled Receptors (GPCR) structures, transmembrane helices, membrane molecular dynamics (MD) simulations, pH-dependent protonation states, membrane permeability, and passive 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 tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Computational Design of Membrane Proteins aims to ensure successful results in the further study of this vital field.Chapter 4 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.
Percolation theory describes the effects of the connectivity of microscopic or small-scale elements of a complex medium to its macroscopic or large-scale properties. It also describes the conditions under which there may be a continuously connected path of local elements across the medium. The point at which the path is formed is called the percolation threshold. Percolation theory also predicts that many macroscopic properties of complex media follow universal power laws near the percolation threshold that are independent of many microscopic features of such media.There are many applications of percolation theory across the natural sciences, from porous materials, to composite solids, complex networks, and biological systems. This book presents the essential elements of percolation theory, covers the problem of calculating the exponents that characterize the power laws that the percolation quantities follow near the percolation threshold, provides a clear description of the geometry of percolation clusters of the connected paths, and addresses several variations of percolation theory. In particular, bootstrap percolation, explosive percolation, and invasion percolation are featured, which expand the range of natural systems to which percolation may be applicable. In addition, coverage includes several important applications of percolation theory to a range of phenomena, ranging from electrical conductivity, thermopower, the Hall effect, and photoconductivity of disordered semiconductors, to flow, transport and reaction in porous media, geochemistry, biology, and ecology.
This book provides basic information and different protocols associated with the Entomopathogenic nematodes (EPNs) and their symbiotic bacteria. Entomopathogenic nematodes (EPNs) of the genera Steinernema and Heterorhabditis and their associated bacterial symbionts Xenorhabdus and Photorhabdus aid nematode infective juveniles (IJs) in infecting and killing their insect hosts, creating a unique tripartite complex of host-vector-symbiont interactions. Due to this insect killing capability, EPNs are used as biological control agents of economically important insect pests. They are also a model system to study host-parasite interactions.It provides a systematic approach to various nematode procedures including pathogenicity, reproduction, foraging behaviour. It gives a brief outline on historical aspects, nematode-bacterium complex, biology and chemical ecology of EPNs. It concisely describes host insect rearing, nematode samplingand storage, isolation techniques, counting, handling and staining of nematodes, characterization including morphological, molecular and ecological studies, mass production, virulence bioassay, field application and efficacy. The book also includes methods and techniques for their associated symbiotic bacteria.This book serves as a laboratory manual and assists the readers to undertake advanced research in different aspects related to nematodes. It is useful for researchers in the fields of nematology, microbiology, bacteriology and entomology.
This second edition volume details the latest aspects of neural cells covering the practical and theoretical considerations of each techniques involved. Chapters guide readers through a general overview of the neuronal culturing principles, cell line models for neural cells, the isolation and propagation of primary cultures, stem cells, transfection and transduction of neural cultures, and other more advanced 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. Practical and easy to use, Neuronal Cell Culture: Methods and Protocols, Second Edition aims to be of interest to scientists at all levels studying cell culture models for neuroscientific studies.
This volume presents the most useful laboratory protocols in strigolactones(SL) research. Chapters guide readers through wet-lab paths, issues around stability, protocols to evaluate SL activity, effects towards soil inhabitants such as parasitic plants, mycorrhizal and non-mycorrhizal fungi, nodulating bacteria, and protocols to assess effects on plant development are discussed. 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, Strigolactones: Methods and Protocols aims to deliver a clear-cut and standardized set of experimental protocols to a broad scientific community.
This volume provides a collection of protocols for the most common experimental methods used for engineering Yarrowia lipolytica. Chapters detail the basic theories underlying the methods described in each chapter. 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, Yarrowia lipolytica: Methods and Protocols aims to provide experimentalists with a full account of the practical steps necessary for carrying out each protocol successfully.
This detailed volume covers conventional MS-based ¿shotgun lipidomics¿ by which samples are introduced by infusion or loop injection, as well as LC-MS-based lipidomics, which are becoming increasingly important due to the ever-increasing demand for a complete and precise lipid analysis of the complex and diversified lipids in nature. The volume features protocols applying chemical reactions, the on-line photochemical reactions combined with various MS methods for comprehensive characterization of various lipid classes, and quantification of specific and rare lipids. 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, Mass Spectrometry-Based Lipidomics: Methods and Protocols serves as an invaluable guide for biochemists and mass spectroscopists who are interested in lipid studies.
This updated and expanded volume reflects the current state of the structural protein field with improved and refined protocols that have been applied to particularly challenging proteins. Beginning with a section on structural bioinformatics, the book continues with sections covering the challenge of producing high quality samples for structural studies, particularly mammalian membrane proteins and protein complexes, as well as protocols for structure determination, including the use of electrons in structural biology and more. 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, Structural Proteomics: High-Throughput Methods, Third Edition will aid researchers in expanding our knowledge of this vital and expansive area of protein science.Chapter 9 is available open access under a CC BY 4.0 license.
This volume provides a wide range of imaging protocols that can be tailored to specific organisms or cell-types. Chapters guide readers through fixed-cell, live-cell, phenotype screening, super-resolution, intravital imaging techniques, and fluorescence life-time imaging microscopy (FLIM). 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, Confocal Microscopy: Methods and Protocols aims to ensure successful results in the further study of this vital field.
This second edition volume expands on the previous edition with new information on the structural analysis of glycosaminoglycans (GAGs); chemical and enzymatic synthesis of GAGs; biophysical, biochemical, and computational analysis of GAG-protein interactions; molecular and genetic approaches to manipulating GAG cell biology; and molecular and genetic methods involving animal models and organoids. This volume also offers insight and guidance to non-glycoscience researchers who are performing advanced experiments on GAGs. 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, Glycosaminoglycans: Methods and Protocols, Second Edition is a valuable experimental manual for practicing researchersincluding graduate and post-doctoral fellows, and provides them with the latest methodologies and practical tips to overcoming barriers in understanding the chemistry and biology of GAGs.
This book examines detailed experimental and computational approaches for the analysis of many aspects vital to the understanding of membrane protein structure and function. Readers will receive guidance on the selection and use of methods for over-expression and purification, tools to characterize membrane proteins within different phospholipid bilayers, direction on functional studies, and approaches to determine the structures of membrane proteins. Detailed experimental steps for specific membrane proteins with critical notes allow the protocols to be modified to different systems. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of practical information and implementation advice that leads to excellent, reproducible results. Authoritative and up-to-date, Structure and Function Studies of Membrane Proteins serves as an ideal guide for biologists, biochemists, and biophysicists striving to further understand these essential proteins and their many biological roles.
This volume details a comprehensive set of methods and tools for Hi-C data processing, analysis, and interpretation. Chapters cover applications of Hi-C to address a variety of biological problems, with a specific focus on state-of-the-art computational procedures adopted for the data analysis. 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, Hi-C Data Analysis: Methods and Protocols aims to help computational and molecular biologists working in the field of chromatin 3D architecture and transcription regulation.
This second edition provides new and updated chapters detailing methods tackling all aspects of small non-coding RNAs biology. Chapters guide readers through customized dedicated protocols and technologies that will be of valuable help decipher the numerous functions of small non-coding RNAs. Written in the highly successful Methods of 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.Instructive and practical, Small Non-Coding RNAs: Methods and Protocols, Second Edition aims to be a valuable help and of great interest to a large number of researchers working in the field of RNA or willing to develop RNA studies.
This detailed book explores the fundamentals of myofibroblast biology in tissue repair, fibrosis, and tumors as well as providing cutting-edge laboratory methods used to investigate myofibroblast functions in physiological and pathological settings in vitro and in vivo, as written by leading academic scientists. Section I of this volume focuses on fundamental methods to study myofibroblast biology and covers topics such as methods for detecting myofibroblasts and senescent myofibroblasts in cell culture and histology, single cell RNA sequencing to identify myofibroblast subsets in fibrotic tissues, and functional assays to assess TGF-¿ activation, myofibroblast apoptosis, or matrix deposition and crosslinking. Section II discusses methods to investigate the mechanobiology of myofibroblasts in vitro, including the fabrication of functional hydrogels with tunable stiffness, the use of atomic force microscopy to characterize matrix and cellular stiffness, as well as molecular assays to assess fibroblast mechanotransduction pathways and durotaxis. Section III describes multiple animal models to investigate myofibroblast functions across organs in vivo as well as human organoid systems, precision tissue slices and decellularized 3D tissue scaffolds to assess myofibroblast functions in relevant human ex vivo 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and extensive, Myofibroblasts: Methods and Protocols is an essential collection for researchers delving into the processes and effects of these important cells.
This detailed book describes some of the most recent advances and up-to-date methodologies to detect, quantify, analyze, and elucidate the biological function of different types of RNA modifications. Importantly, the methodologies and tools described herein can be applied to a wide variety of organisms and can be used to address biological and clinical questions. Beginning with a section on bioinformatics tools, the collection continues with sections on detecting RNA modifications using Nanopore direct RNA sequencing, next-generation sequencing approaches, qPCR- and molecular biology-based methods, mass spectrometry- and NMR-based methods, as well as approaches to assess kinetics, determinants, and functions of RNA modifications. Written for the highly successful Methods in Molecular Biology series style, chapters include introduction 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, RNA Modifications: Methods and Protocols serves as an ideal guide for those working directly in the fields of epitranscriptomics and post-transcriptional gene regulation, as well as for scientists and clinicians interested in bioinformatic tools to study RNA modifications and techniques to dissect their roles in physiology and disease.Chapter 20 is available open access under a CC BY 4.0 license.
This essential volume explores a variety of tools and protocols of structure-based (homology modeling, molecular docking, molecular dynamics, protein-protein interaction network) and ligand-based (pharmacophore mapping, quantitative structure-activity relationships or QSARs) drug design for ranking and prioritization of candidate molecules in search of effective treatment strategy against coronaviruses. Beginning with an introductory section that discusses coronavirus interactions with humanity and COVID-19 in particular, the book then continues with sections on tools and methodologies, literature reports and case studies, as well as online tools and databases that can be used for computational anti-coronavirus drug research. Written for the Methods in Pharmacology and Toxicology series, chapters include the kind of practical detail and implementation advice that ensures high quality results in the lab. Comprehensive and timely, In Silico Modeling ofDrugs Against Coronaviruses: Computational Tools and Protocols is an ideal reference for researchers working on the development of novel anti-coronavirus drugs for SARS-CoV-2 and for coronaviruses that will likely appear in the future.
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