In the last few years, significant advances have been made in understanding how a yeast cell responds to the stress of producing a recombinant protein, and how this information can be used to engineer improved host strains. The molecular biology of the expression vector, through the choice of promoter, tag and codon optimization of the target gene, is also a key determinant of a high-yielding protein production experiment. Recombinant Protein Production in Yeast: Methods and Protocols examines the process of preparation of expression vectors, transformation to generate high-yielding clones, optimization of experimental conditions to maximize yields, scale-up to bioreactor formats and disruption of yeast cells to enable the isolation of the recombinant protein prior to purification. Written in the highly successful Methods in Molecular Biology (TM) 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 practical, Recombinant Protein Production in Yeast: Methods and Protocols, seeks to aid scientists in adopting yeast as a protein production host.

Computational Biomechanics for Medicine: Solid and fluid mechanics for the benefit of patients contributions and papers from the MICCAI Computational Biomechanics for Medicine Workshop help in conjunction with Medical Image Computing and Computer Assisted Intervention conference (MICCAI 2020) in Lima, Peru. The content is dedicated to research in the field of methods and applications of computational biomechanics to medical image analysis, image-guided surgery, surgical simulation, surgical intervention planning, disease prognosis and diagnostics, analysis of injury mechanisms, implant and prostheses design, as well as artificial organ design and medical robotics. This book appeals to researchers, students and professionals in the field.

Flow cytometry forms an integral part of both basic biological research and clinical diagnosis in pathology. This straightforward new volume provides a clear, easy-to-read, and practical manual for both clinicians and non-clinicians at all levels of their careers.

The chapter topics range from basic principles to more advanced subjects, such as apoptosis and cell sorting. The book charts the history, development and basic principles of flow cytometry.

This book summarizes the main surface analysis techniques that are being used to study biological specimens/systems. The compilation of chapters in this book highlight the benefits that surface analysis provides.

The outer layer of bulk solid or liquid samples is referred to as the surface of the sample/material. At the surface, the composition, microstructure, phase, chemical bonding, electronic states, and/or texture is often different than that of the bulk material. The outer surface is where many material interactions/reactions take place. This is especially true biomaterials which may be fabricated into bio-devices and in turn implanted into tissues and organs. Surfaces of biomaterials (synthetic or modified natural materials) are of critical importance since the surface is typically the only part of the biomaterial/bio-device that comes in contact with the biological system.

Analytical techniques are required to characterize the surface of biomaterials and quantify their impact in real-world biological systems. Surface analysis of biological materials started in the 1960 s and the number of researchers working in this area have increased very rapidly since then, a number of advances have been made to standard surface analytical instrumentation, and a number of new instruments have been introduced.

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International Review of Cytology presents current advances and comprehensive reviews in cell biology both plant and animal. Authored by some of the foremost scientists in the field, each volume provides up-to-date information and directions for future research. Articles in this volume address endogenous ligands of PACAP, VIP receptors in the autocrine-paracrine regulation of the adrenal gland; ultrastructural dynamcs of human reproduction, from ovulation to fertilization and early embryo development; chromosomal variation in mammalian neuronal cells; automated interpretation of protein subcellular location patterns; cell and molecular biology of human lacrimal gland and nasolacrimal duct mucins.

This book presents recent methods for Systems Genetics (SG) data analysis, applying them to a suite of simulated SG benchmark datasets. Each of the chapter authors received the same datasets to evaluate the performance of their method to better understand which algorithms are most useful for obtaining reliable models from SG datasets. The knowledge gained from this benchmarking study will ultimately allow these algorithms to be used with confidence for SG studies e.g. of complex human diseases or food crop improvement. The book is primarily intended for researchers with a background in the life sciences, not for computer scientists or statisticians.

The proposed volume provides both fundamental and detailed information about the computational and computational-experimental studies which improve our knowledge of how leaving matter functions, the different properties of drugs (including the calculation and the design of new ones), and the creation of completely new ways of treating numerical diseases. Whenever it is possible, the interplay between theory and experiment is provided. The book features computational techniques such as quantum-chemical and molecular dynamic approaches and quantitative structure-activity relationships. The initial chapters describe the state-of-the art research on the computational investigations in molecular biology, molecular pharmacy, and molecular medicine performed with the use of pure quantum-chemical techniques. The central part of the book illustrates the status of computational techniques that utilize hybrid, so called QM/MM approximations as well as the results of the QSAR studies which now are the most popular in predicting drugs' efficiency. The last chapters describe combined computational and experimental investigations.

This thorough volume provides an in-depth introduction to and discussion of microRNAs (miRs) and their targets, miR functions, and computational techniques applied in miR research, thus serving the need for a comprehensive book focusing on miR target genes, miR regulation mechanisms, miR functions performed in various human diseases, and miR databases/knowledgebases. Without prior knowledge of the area of study, computational biologists, computer scientists, bioinformaticians, bench biologists, as well as clinical investigators will find it easy to follow the techniques in this collection. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detailed implementation advice that ensures successful results. Accessible and practical, Bioinformatics in MicroRNA Research functions as an ideal guide for researchers of all backgrounds to explore this vital area of study.

Thisbookisintendedformolecularbiologistswhoperformquantitativeanalysesondata emanatingfromtheir?eldandforthestatisticianswhoworkwithmolecularbiologists andotherbiomedicalresearchers. Therearemanyexcellenttextbooksthatprovidefun- mentalcomponentsforstatisticaltrainingcurricula. Therearealsomany"byexpertsfor experts"booksinstatisticsandmolecularbiologywhichrequirein-depthknowledgein bothsubjectstobetakenfulladvantageof. Sofar,nobookinstatisticshasbeenpublished thatprovidesthebasicprinciplesofproperstatisticalanalysesandprogressestoamore advancedstatisticsinresponsetorapidlydevelopingtechnologiesandmethodologiesin the?eldofmolecularbiology. Respondingtothissituation,ourbookaimsatbridgingthegapbetweenthesetwo extremes. Molecularbiologistswillbene?tfromtheprogressivestyleofthebookwhere basicstatisticalmethodsareintroducedandgraduallyelevatedtoanintermediatelevel. Similarly,statisticianswillbene?tfromlearningthevariousbiologicaldatageneratedfrom the?eldofmolecularbiology,thetypesofquestionsofinteresttomolecularbiologists, andthestatisticalapproachestoanalyzingthedata. Thestatisticalconceptsandmethods relevanttostudiesinmolecularbiologyarepresentedinasimpleandpracticalmanner. Speci?cally,thebookcoversbasicandintermediatestatisticsthatareusefulforclassical and molecular biology settings and advanced statistical techniques that can be used to helpsolveproblemscommonlyencounteredinmodernmolecularbiologystudies,such assupervisedandunsupervisedlearning,hiddenMarkovmodels,manipulationandan- ysisofdatafromhigh-throughputmicroarrayandproteomicplatform,andsynthesisof these evidences. A tutorial-type format is used to maximize learning in some chapters. Advicefromjournaleditorsonpeer-reviewedpublicationandsomeusefulinformationon softwareimplementationarealsoprovided. Thisbookisrecommendedforuseassupplementarymaterialbothinsideandoutside classroomsorasaself-learningguideforstudents,scientists,andresearcherswhodealwith numericdatainmolecularbiologyandrelated?elds. Thosewhostartasbeginners,but desiretobeatanintermediatelevel,will?ndthisbookespeciallyusefulintheirlearning pathway. WewanttothankJohnWalker(serieseditor),PatrickMarton,DavidCasey,andAnne Meagher,(editorsatSpringerandHumana)andShanthyJaganathan(Integra-India). The followingpersonsprovidedusefuladviceandcommentsonselectionoftopics,referralto expertsineachtopic,and/orchapterreviewsthatwetrulyappreciate:StephenLooney(a former editor of this book), Stan Young, Dmitri Zaykin, Douglas Hawkins, Wei Pan, Alexandre Almeida, John Ho, Rebecca Doerge, Paula Trushin, Kevin Morgan, Jason Osborne,PeterWestfall,JennyXiang,Ya-linChiu,YolandaBarron,HuiboShao,Alvin Mushlin,andRonaldFanta. Drs. Bang,Zhou,andMazumdarwerepartiallysupported byClinicalTranslationalScienceCenter(CTSC)grant(UL1-RR024996). HeejungBang vii Contents Preface...vii Contributors...xi PARTIBASICSTATISTICS...1 1. ExperimentalStatisticsforBiologicalSciences...3 HeejungBangandMarieDavidian 2. NonparametricMethodsforMolecularBiology...105 KnutM. WittkowskiandTingtingSong 3. BasicsofBayesianMethods...155 SujitK. Ghosh 4. TheBayesiant-TestandBeyond ...179 MithatGonen PARTII DESIGNSANDMETHODSFORMOLECULARBIOLOGY...201 5. SampleSizeandPowerCalculationforMolecularBiologyStudies...203 Sin-HoJung 6. DesignsforLinkageAnalysisandAssociationStudiesofComplexDiseases...219 YuehuaCui,GengxinLi,ShaoyuLi,andRonglingWu 7. IntroductiontoEpigenomicsandEpigenome-WideAnalysis...243 MelissaJ. FazzariandJohnM. Greally 8. Exploration,Visualization,andPreprocessingofHigh-DimensionalData...267 ZhijinWuandZhiqiangWu PARTIII STATISTICALMETHODSFORMICROARRAYDATA ...285 9. IntroductiontotheStatisticalAnalysisofTwo-ColorMicroarrayData...287 MartinaBremer,EdwardHimelblau,andAndreasMadlung 10. BuildingNetworkswithMicroarrayData...315 BradleyM. Broom,WareeRinsurongkawong,LajosPusztai, andKim-AnhDo PARTIV ADVANCEDORSPECIALIZEDMETHODSFORMOLECULARBIOLOGY. . 345 11. SupportVectorMachinesforClassi?cation:AStatisticalPortrait...347 YoonkyungLee 12. AnOverviewofClusteringAppliedtoMolecularBiology ...369 RebeccaNugentandMarinaMeila ix xContents 13. HiddenMarkovModelandItsApplicationsinMotifFindings...405 JingWuandJunXie 14. DimensionReductionforHigh-DimensionalData...417 LexinLi 15. IntroductiontotheDevelopmentandValidationofPredictiveBiomarker ModelsfromHigh-ThroughputDataSets ...435 XutaoDengandFabienCampagne 16. Multi-geneExpression-basedStatisticalApproachestoPredicting Patients'ClinicalOutcomesandResponses...471 FengCheng,Sang-HoonCho,andJaeK. Lee 17. Two-StageTestingStrategiesforGenome-WideAssociationStudies inFamily-BasedDesigns ...485 AmyMurphy,ScottT. Weiss,andChristophLange 18. StatisticalMethodsforProteomics ...497 KlausJung PARTVMETA-ANALYSISFORHIGH-DIMENSIONALDATA ...509 19. StatisticalMethodsforIntegratingMultipleTypesofHigh-ThroughputData. . 511 YangXieandChulAhn 20. ABayesianHierarchicalModelforHigh-DimensionalMeta-analysis...531 FeiLiu 21. MethodsforCombiningMultipleGenome-WideLinkageStudies...541 TreciaA. KippolaandStephanieA. Santorico PARTVI OTHERPRACTICALINFORMATION ...561 22. ImprovedReportingofStatisticalDesignandAnalysis:Guidelines, Education,andEditorialPolicies...5 63 MadhuMazumdar,SampritBanerjee,andHeatherL. VanEpps 23. StataCompanion...599 JenniferSousaBrennan SubjectIndex...627 Contributors CHULAHN* Division of Biostatistics, Department of Clinical Sciences, The Harold C.

This second edition expands upon the previous volume with additional emphasis on recent innovation in basic renal research and has a more holistic approach on associated disorders such as complications associated with peritoneal dialysis, ischemic acute kidney injury, the sympathetic nervous system, and vascular calcification. The book is divided into five parts: Part I provides a number of in vitro, in vivo, and ex vivo models of kidney disease and associated complications; Part II looks at recent advances in imaging techniques; Part III covers recent developments in studying metabolism in renal ischemia and reperfusion; Part IV addresses study and measurement of vascular calcification; and Part V explores analytical techniques that are both topical and of widespread relevance to the study of experimental renal disease. 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, Kidney Research: Experimental Protocols, Second Edition is a valuable collection of protocols useful to new and experienced researchers who are interested in the field of Nephrology.

The field of bacterial diagnostics has seen unprecedented advances in recent years. The increased need for accurate detection and identification of bacteria in human, animal, food, and environmental samples has fueled the development of new techniques. The field has seen extensive research aided by the information from bacterial genome sequencing projects. Although traditional methods of bacterial detection and identification remain in use in laboratories around the world, there is now a growing trend toward the use of nucleic ac- based diagnostics and alternative biochemically and immunologically based formats. The ultimate goal of all diagnostic tests is the accurate detection, identification, or typing of microorganisms in samples of interest. Although the resulting information is of obvious use in the areas of patient management, animal health, and quality control, it is also of use in monitoring routes of infection and outlining strategies for infection control. There is, therefore, a need to ensure that the information being provided is of the highest standard and that any new technique is capable of delivering this.

Leading chemists and engineers concisely explain the principles behind microchip capillary electrophoresis and demonstrate its use in a variety of biochemical applications, ranging from the analysis of DNA, proteins, and peptides to single cell analysis and measuring the impact of surface modification on flow in microfluidic channels. Since surface chemistry must be carefully considered for optimal operation at this scale, the authors also discuss methods of both adsorbed and covalent surface modification for its control. Fabrication methods for producing microchips with glass, poly(dimethylsiloxane), and other polymers are also provided so that even novices can produce simple devices for standard separations. Microchip Capillary Electrophoresis: Methods and Protocols provides a practical starting point for either initiating research in the field of microchip capillary electrophoresis or understanding the full range of what can be done with existing systems.

In this book, seven chapters describe studies aimed at understanding and exploiting the key features of such molecular RNA and DNA devices. In the first section of the book, four chapters are devoted to artificial nucleic acid switches and sensors. These chapters introduce the concept of allosteric ribozymes as molecular switches and sensors; describe nucleic acid enzymes that are switched by oligonucleotides and other nucleic acid enzymes that are switched by proteins; and illustrate how switching elements can be integrated rationally into fluorescently signaling molecular sensors made out of nucleic acids. In the second section of the book, three chapters show that nature has been as crafty a molecular-scale engineer as any modern scientist via evolution of natural nucleic acid switches and sensors. RNAs have been found whose activities are modulated either by proteins or by small-molecule metabolites, and both kinds of system are described. Finally, the notion of exploiting naturally occurring RNA switches for drug development is discussed.

This book brings together the most important research developments of the past 45 years that have enriched our knowledge and contributed to a better understanding of the biochemistry and cell and molecular biology of basement membranes. It describes the studies that shed light on the ultrastructural organization, the biosynthesis of the macromolecular components, their functions in embryonic development and differentiation, and in the mature state. A major portion of the book is devoted to the description of the genes that regulate the expression of the various structural macromolecules.
* Reviews the early years of research and the discovery of type IV collagen
* Presents the diversity of basement membrane morphology
* Discusses gene structure

Goringer 's brilliant new work dedicates a chapter to each of the main types of RNA editing the very first volume to do so. All of the sections here have been written by experts in the various research areas and a specific focus is put on the correlation between RNA structure and function, as well as on the complex cellular machineries that catalyze the different editing reactions. This leads to a "state of the art" compendium of our current knowledge on RNA editing.

High throughput sequencing (HTS) technologies have conquered the genomics and epigenomics worlds. The applications of HTS methods are wide, and can be used to sequence everything from whole or partial genomes, transcriptomes, non-coding RNAs, ribosome profiling, to single-cell sequencing. Having such diversity of alternatives, there is a demand for information by research scientists without experience in HTS that need to choose the most suitable methodology or combination of platforms and to define their experimental designs to achieve their specific objectives. Field Guidelines for Genetic Experimental Designs in High-Throughput Sequencing aims to collect in a single volume all aspects that should be taken into account when HTS technologies are being incorporated into a research project and the reasons behind them. Moreover, examples of several successful strategies will be analyzed to make the point of the crucial features. This book will be of use to all scientist that are unfamiliar with HTS and want to incorporate such technologies to their research.

Sheds new light on intrinsically disordered proteins and peptides, including their role in neurodegenerative diseases

With the discovery of intrinsically disordered proteins and peptides (IDPs), researchers realized that proteins do not necessarily adopt a well defined secondary and tertiary structure in order to perform biological functions. In fact, IDPs play biologically relevant roles, acting as inhibitors, scavengers, and even facilitating DNA/RNA-protein interactions. Due to their propensity for self-aggregation and fibril formation, some IDPs are involved in neurodegenerative diseases such as Parkinson's and Alzheimer's.

With contributions from leading researchers, this text reviews the most recent studies, encapsulating our understanding of IDPs. The authors explain how the growing body of IDP research is building our knowledge of the folding process, the binding of ligands to receptor molecules, and peptide self-aggregation. Readers will discover a variety of experimental, theoretical, and computational approaches used to better understand the properties and function of IDPs. Moreover, they'll discover the role of IDPs in human disease and as drug targets.

Protein and Peptide Folding, Misfolding, and Non-Folding begins with an introduction that explains why research on IDPs has significantly expanded in the past few years. Next, the book is divided into three sections:

Conformational Analysis of Unfolded States

Disordered Peptides and Molecular Recognition

Aggregation of Disordered Peptides

Throughout the book, detailed figures help readers understand the structure, properties, and function of IDPs. References at the end of each chapter serve as a gateway to the growing body of literature in the field.

With the publication of Protein and Peptide Folding, Misfolding, and Non-Folding, researchers now have a single place to discover IDPs, their diverse biological functions, and the many disciplines that have contributed to our evolving understanding of them.

Nucleic acids are the fundamental building blocks of DNA and RNA and are found in virtually every living cell. Molecular biology is a branch of science that studies the physicochemical properties of molecules in a cell, including nucleic acids, proteins, and enzymes. Increased understanding of nucleic acids and their role in molecular biology will further many of the biological sciences including genetics, biochemistry, and cell biology. Progress in Nucleic Acid Research and Molecular Biology is intended to bring to light the most recent advances in these overlapping disciplines with a timely compilation of reviews comprising each volume.

"Next generation" sequencing techniques allow for more detailed analysis of exons and introns in multiple genes at the same time. This will reveal many mutations that potentially lead to exon skipping. To functionally test these a lot can be achieved with a limited set of protocols, while for the intentional induction of exon skipping different tools and target genes are involved and the translational path from in vitro splicing to in vivo tests in animal models requiring a more extensive set of protocols. Exon Skipping: Methods and Protocols provides scientist with a comprehensive guide to many of the methods and techniques used for exon skipping, such as methods on how to discriminate "real polymorphisms" from mutations that affect splicing. Written in the highly successful Methods in Molecular Biology (TM) 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 practical Exon Skipping: Methods and Protocols seeks to aid scientists in the continuing study of exon skipping.

This volume arranged into three sections describes biochemical, in vitro, and in vivo protocols on Semaphorins. Chapters focus on approaches that would allow the novice to study Semaphorins and employ robust assays to characterize mechanisms of action. 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, Semaphorin Signaling: Methods and Protocols aims to ensure successful results in the further study of this vital field.

What Arieh Warshel and fellow 2013 Nobel laureates Michael Levitt and Martin Karplus achieved - beginning in the late 1960s and early 1970s when computers were still very primitive - was the creation of methods and programs that describe the action of biological molecules by 'multiscale models'. In this book, Warshel describes this fascinating, half-century journey to the apex of science.From Kibbutz Fishponds to The Nobel Prize is as much an autobiography as an advocacy for the emerging field of computational science. We follow Warshel through pivotal moments of his life, from his formative years in war-torn Israel in an idealistic kibbutz that did not encourage academic education; to his time in the army and his move to the Technion where he started in his obsession of understanding the catalytic power of enzymes; to his eventual scientific career which took him to the Weizmann Institute, Harvard University, Medical Research Council, and finally University of Southern California. We read about his unique contributions to the elucidation of the molecular basis of biological functions, which are combined with instructive stories about his persistence in advancing ideas that contradict the current dogma, and the nature of his scientific struggle for recognition, both personal and for the field to which he devoted his life. This is, in so many ways, more than just a memoir: it is a profoundly inspirational tale of one man's odyssey from a kibbutz that did not allow him to go to a university to the pinnacle of the scientific world, highlighting that the correct mixture of persistence, talent and luck can lead to a Nobel Prize.

Mass Spectroscopy Imaging (MSI) has emerged as an enabling technique to provide insight into the molecular entities within cells, tissues and whole-body samples and to understand inherent complexities within biological metabolomes. In "Mass ""Spectrometry"" Imaging of Small Molecules"" Methods and Protocols," experts in the MSI field present techniques for 2D and 3D visualization and quantification of a wide array of small molecular species present in biologically relevant samples. Chapters provide detailed operational instructions from sample preparation to method selection, from comparative quantification to structural identification and from data collection to visualization of small molecule mapping in complex samples. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, "Mass ""Spectrometry"" Imaging of Small Molecules"" Methods and Protocols" aims to bring the rapidly maturing methods of metabolic imaging to life science researchers and to minimize technical intimidation in adapting new technological platforms in biological research.

This volume discusses various basic and advanced methods and protocols that have been proven to be successful among certain bacterial species, or a family of species, in type III secretion systems (T3S system). The chapters in this book cover topics such as: site-directed mutagenesis and its application in studying the interactions of T3S components; use of transcriptional control to increase expression and secretion of heterologous proteins in T3S systems; fractionation techniques to examine effector translocation; detecting immune responses to T3S systems; mouse immunization with purified needle proteins from T3S systems and the characterization of the immune responses to these proteins; and detection of protein interactions in T3S systems using yeast 2-hybrid 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. Practical and comprehensive, Type 3 Secretion Systems: Methods and Protocols is a valuable resource for anyone interested in learning about the fascinating and ever-changing T3S systems.

The intersection of nanotechnology with biology has given rise to numerous ideas for new ways to use nanotechnology for biological applications. Nanomaterials possess unique size- and material-dependent properties which make them attractive for improving regular biomedical fields, such as drug delivery, imaging, therapy, and diagnostics. Divided into three convenient sections, Nanomaterial Interfaces in Biology: Methods and Protocols covers protocols describing synthesis, fabrication, and construction of bio-nanomaterial interfaces, characterization protocols of bio-nanomaterial interfaces, and applications which utilize the bio-nanomaterial interfaces. Written in the highly successful Methods in Molecular Biology series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible Nanomaterial Interfaces in Biology: Methods and Protocols will serve the new and emerging scientific community, enabling new capabilities and technologies that were not previously possible in medicine and biology.