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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.

This second edition further develops the principles of applying kinetic principles to drug metabolizing enzymes and transporters. Chapters are divided into six sections detailing fundamental principles of enzyme kinetics, enzyme and transporter structures, highlighting specific oxidative and conjugative drug metabolizing enzymes and drug transporters, modeling approaches for drug metabolizing enzymes and transporters, understanding of variability both experimental and interindividual (pharmacogenomic), and expanded case studies that provide real life examples of applying these principles. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, in some cases step-by-step instructions with readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls with extensive cross referencing to assist in learning. Authoritativeand fully updated, Enzyme Kinetics in Drug Metabolism: Fundamentals and Applications, Second Edition serves as a practical teaching tool for novice and advanced scientists interested in the fundamental concepts.

This collection explores detailed experimental protocols necessary for setting up a variety of in vitro cytochrome P450 (CYP) assays that are vital in selecting drug candidates in a drug discovery pipeline. Major factors affecting drug metabolism include CYP expression levels, kinetic parameters for individual CYP enzymes, CYP inhibition and induction, time-dependent inhibition (TDI), CYP stability, non-CYP stability, UDP-glucuronosyltransferases (UGT) stability, excretion mechanisms, and drug-drug interactions (DDI), all addressed in this volume. Written for the Methods in Pharmacology and Toxicology series, chapters include helpful background information on the in vitro assay, a list of all the materials, reagents, and equipment necessary to carry out the assay, a step-by-step protocol, notes containing common and unexpected experimental problems in the assay, as well as references containing important supplementary reading. Authoritativeand practical, Cytochrome P450: In Vitro Methods and Protocols serves as a key guide for researchers in the area of discovery and development of new medicines.

This volume provides a comprehensive collection of methods for plant breeders and researchers working in functional genomics of cereal crops. Chapters detail advances in sequencing of cereal genomes, methods of traditional plant breeding, use of machine learning for genomic selection, random and targeted mutagenesis with CRISPR/Cas9, quantitative proteomics and phenotyping in cereals. Authoritative and cutting-edge, Accelerated Breeding of Cereal Crops aims to be of interest to plant breeders, researchers, postdoctoral fellows, and students working in functional genomics for the development of the next generation of crop plants.

This new edition explores lab protocols describing new techniques to study cytotoxic T-cells (CTLs), as well as chapters of a more general discursive nature, all with an emphasis on the use of systems biology in immunology. Beginning with phenotypical characterization of CTL populations, the volume continues with in vitro and in vivo cytotoxicity assays, methods to detect senescent T cells, in vivo and in vitro models to understand immune and bone cells cross-talk, microscopy and in vivo imaging, as well as "Omics" approaches and molecular methods, concluding with chapters on CTL involvement in transplantation and link microbiota-immunity. Written for the highly successful Methods in Molecular Biology series, chapters feature the kind of detail and key implementation advice for best results in the lab. Authoritative and up-to-date, Cytotoxic T-Cells: Methods and Protocols, Second Edition serves as an ideal guide for researchers working with these vital cells.

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.