Computational And Statistical Methods For Protein Quantification By Mass Spectrometry

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Computational and Statistical Methods for Protein Quantification by Mass Spectrometry

Author : Ingvar Eidhammer
Publisher : John Wiley & Sons
ISBN 13 : 111849377X
Total Pages : 290 pages
Book Rating : 4.1/5 (184 download)

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Book Synopsis Computational and Statistical Methods for Protein Quantification by Mass Spectrometry by : Ingvar Eidhammer

Download or read book Computational and Statistical Methods for Protein Quantification by Mass Spectrometry written by Ingvar Eidhammer and published by John Wiley & Sons. This book was released on 2012-12-10 with total page 290 pages. Available in PDF, EPUB and Kindle. Book excerpt: The definitive introduction to data analysis in quantitative proteomics This book provides all the necessary knowledge about mass spectrometry based proteomics methods and computational and statistical approaches to pursue the planning, design and analysis of quantitative proteomics experiments. The author’s carefully constructed approach allows readers to easily make the transition into the field of quantitative proteomics. Through detailed descriptions of wet-lab methods, computational approaches and statistical tools, this book covers the full scope of a quantitative experiment, allowing readers to acquire new knowledge as well as acting as a useful reference work for more advanced readers. Computational and Statistical Methods for Protein Quantification by Mass Spectrometry: Introduces the use of mass spectrometry in protein quantification and how the bioinformatics challenges in this field can be solved using statistical methods and various software programs. Is illustrated by a large number of figures and examples as well as numerous exercises. Provides both clear and rigorous descriptions of methods and approaches. Is thoroughly indexed and cross-referenced, combining the strengths of a text book with the utility of a reference work. Features detailed discussions of both wet-lab approaches and statistical and computational methods. With clear and thorough descriptions of the various methods and approaches, this book is accessible to biologists, informaticians, and statisticians alike and is aimed at readers across the academic spectrum, from advanced undergraduate students to post doctorates entering the field.

Statistical Analysis of Proteomics, Metabolomics, and Lipidomics Data Using Mass Spectrometry

Author : Susmita Datta
Publisher : Springer
ISBN 13 : 3319458094
Total Pages : 295 pages
Book Rating : 4.3/5 (194 download)

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Book Synopsis Statistical Analysis of Proteomics, Metabolomics, and Lipidomics Data Using Mass Spectrometry by : Susmita Datta

Download or read book Statistical Analysis of Proteomics, Metabolomics, and Lipidomics Data Using Mass Spectrometry written by Susmita Datta and published by Springer. This book was released on 2016-12-15 with total page 295 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book presents an overview of computational and statistical design and analysis of mass spectrometry-based proteomics, metabolomics, and lipidomics data. This contributed volume provides an introduction to the special aspects of statistical design and analysis with mass spectrometry data for the new omic sciences. The text discusses common aspects of design and analysis between and across all (or most) forms of mass spectrometry, while also providing special examples of application with the most common forms of mass spectrometry. Also covered are applications of computational mass spectrometry not only in clinical study but also in the interpretation of omics data in plant biology studies. Omics research fields are expected to revolutionize biomolecular research by the ability to simultaneously profile many compounds within either patient blood, urine, tissue, or other biological samples. Mass spectrometry is one of the key analytical techniques used in these new omic sciences. Liquid chromatography mass spectrometry, time-of-flight data, and Fourier transform mass spectrometry are but a selection of the measurement platforms available to the modern analyst. Thus in practical proteomics or metabolomics, researchers will not only be confronted with new high dimensional data types—as opposed to the familiar data structures in more classical genomics—but also with great variation between distinct types of mass spectral measurements derived from different platforms, which may complicate analyses, comparison, and interpretation of results.

Computational Methods for Mass Spectrometry Proteomics

Author : Ingvar Eidhammer
Publisher : John Wiley & Sons
ISBN 13 : 9780470724293
Total Pages : 296 pages
Book Rating : 4.7/5 (242 download)

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Book Synopsis Computational Methods for Mass Spectrometry Proteomics by : Ingvar Eidhammer

Download or read book Computational Methods for Mass Spectrometry Proteomics written by Ingvar Eidhammer and published by John Wiley & Sons. This book was released on 2008-02-28 with total page 296 pages. Available in PDF, EPUB and Kindle. Book excerpt: Proteomics is the study of the subsets of proteins present in different parts of an organism and how they change with time and varying conditions. Mass spectrometry is the leading technology used in proteomics, and the field relies heavily on bioinformatics to process and analyze the acquired data. Since recent years have seen tremendous developments in instrumentation and proteomics-related bioinformatics, there is clearly a need for a solid introduction to the crossroads where proteomics and bioinformatics meet. Computational Methods for Mass Spectrometry Proteomics describes the different instruments and methodologies used in proteomics in a unified manner. The authors put an emphasis on the computational methods for the different phases of a proteomics analysis, but the underlying principles in protein chemistry and instrument technology are also described. The book is illustrated by a number of figures and examples, and contains exercises for the reader. Written in an accessible yet rigorous style, it is a valuable reference for both informaticians and biologists. Computational Methods for Mass Spectrometry Proteomics is suited for advanced undergraduate and graduate students of bioinformatics and molecular biology with an interest in proteomics. It also provides a good introduction and reference source for researchers new to proteomics, and for people who come into more peripheral contact with the field.

Selected Reaction Monitoring Mass Spectrometry (SRM-MS) in Proteomics

Author : Mahmud Hossain
Publisher : Springer Nature
ISBN 13 : 3030534332
Total Pages : 283 pages
Book Rating : 4.0/5 (35 download)

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Book Synopsis Selected Reaction Monitoring Mass Spectrometry (SRM-MS) in Proteomics by : Mahmud Hossain

Download or read book Selected Reaction Monitoring Mass Spectrometry (SRM-MS) in Proteomics written by Mahmud Hossain and published by Springer Nature. This book was released on 2020-09-26 with total page 283 pages. Available in PDF, EPUB and Kindle. Book excerpt: Covering a wide-ranging facet of a “gold-standard” targeted mass spectrometry (MS) method for the consistent detection and accurate quantification of preselected proteins in complex biological matrices, Selected Reaction Monitoring Mass Spectrometry (SRM-MS) in Proteomics: A Comprehensive View describes: The knowledge-based development of highly efficient SRM methodology including assay workflow, selection of proteins, peptides, transitions and its validation, and quality assessment Available bioinformatic tools – for both pre-acquisition method development and post-MS acquisition data analysis and data repositories Various relative and absolute quantification techniques SRM-MS’ widespread applications in biomarker development and in clinical studies, as well as in the analysis of various posttranslational modifications (PTMs) Current challenges and contemporary trends to overcome those difficulties In addition, it features the historical development of modern-day mass spectrometry with its vivid applications and also covers basic MS instrumentation, ionization techniques, and various proteomics approaches. Comprehensive discussion, extensive references at the end of each chapter, and the list of review articles in the bibliography offer invaluable resources for advanced readings. Researchers from the undergraduate to postgraduate level and beyond in both academic or industry settings studying and working on mass spectrometry and/or proteomics will benefit from this book.

Mass Spectrometry Data Analysis in Proteomics

Author : Rune Matthiesen
Publisher : Humana
ISBN 13 : 9781493997435
Total Pages : 470 pages
Book Rating : 4.9/5 (974 download)

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Book Synopsis Mass Spectrometry Data Analysis in Proteomics by : Rune Matthiesen

Download or read book Mass Spectrometry Data Analysis in Proteomics written by Rune Matthiesen and published by Humana. This book was released on 2019-10-01 with total page 470 pages. Available in PDF, EPUB and Kindle. Book excerpt: The aim of this new edition is to provide detailed information on each topic and present novel ideas and views that can influence future developments in mass spectrometry-based proteomics. In contrast to the previous editions, this third edition aims to provide the most relevant computational methods, focusing on computational concepts. 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, Mass Spectrometry Data Analysis in Proteomics, Third Edition to ensure successful results in the further study of this vital field.

Separation Techniques Applied to Omics Sciences

Author : Ana Valéria Colnaghi Simionato
Publisher : Springer Nature
ISBN 13 : 3030772527
Total Pages : 277 pages
Book Rating : 4.0/5 (37 download)

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Book Synopsis Separation Techniques Applied to Omics Sciences by : Ana Valéria Colnaghi Simionato

Download or read book Separation Techniques Applied to Omics Sciences written by Ana Valéria Colnaghi Simionato and published by Springer Nature. This book was released on 2021-10-09 with total page 277 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book covers liquid chromatography, gas chromatography and capillary electrophoresis, the three main separation techniques lately available, applied to key omic sciences, such as genomics, proteomics, metabolomics and foodomics. The fundamentals of each technique are not covered herein. Instead, the recent advances in such techniques are presented focusing on the application to omics analyses and unique aspects in each case. This volume intends to offer wide ranging options available to researchers on omics sciences, and how to integrate them in order to achieve the comprehension of a biological system as a whole. Omic sciences have been of ultimate importance to comprehend the complex biochemical reactions and related events that occurs upon a biological system. The classical central dogma of molecular biology, which states that genetic information flows unidirectionally from DNA to RNA and then to proteins, has been gradually replaced by the systems biology approach. This book presents a multidisciplinary approach that explains the biological system as a whole, where the entire organism is influenced by a variety of internal events as well as by the environment, showing that each level of the biological information flux may influence the previous or the subsequent one.

Proteomics Data Analysis

Author : Daniela Cecconi
Publisher :
ISBN 13 : 9781071616413
Total Pages : 326 pages
Book Rating : 4.6/5 (164 download)

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Book Synopsis Proteomics Data Analysis by : Daniela Cecconi

Download or read book Proteomics Data Analysis written by Daniela Cecconi and published by . This book was released on 2021 with total page 326 pages. Available in PDF, EPUB and Kindle. Book excerpt: This thorough book collects methods and strategies to analyze proteomics data. It is intended to describe how data obtained by gel-based or gel-free proteomics approaches can be inspected, organized, and interpreted to extrapolate biological information. Organized into four sections, the volume explores strategies to analyze proteomics data obtained by gel-based approaches, different data analysis approaches for gel-free proteomics experiments, bioinformatic tools for the interpretation of proteomics data to obtain biological significant information, as well as methods to integrate proteomics data with other omics datasets including genomics, transcriptomics, metabolomics, and other types of data. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detailed implementation advice that will ensure high quality results in the lab. Authoritative and practical, Proteomics Data Analysis serves as an ideal guide to introduce researchers, both experienced and novice, to new tools and approaches for data analysis to encourage the further study of proteomics.

Quantitative Methods in Proteomics

Author : Katrin Marcus
Publisher :
ISBN 13 : 9781071610244
Total Pages : 483 pages
Book Rating : 4.6/5 (12 download)

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Book Synopsis Quantitative Methods in Proteomics by : Katrin Marcus

Download or read book Quantitative Methods in Proteomics written by Katrin Marcus and published by . This book was released on 2021 with total page 483 pages. Available in PDF, EPUB and Kindle. Book excerpt: This second edition provides new and updated methods on the principles underlying modern protein analysis, from statistical issues to gel-based and mass spectrometry-based applications. Chapters detail protein quantification as basis for realisation of quantitative studies, gel-based and mass spectrometry-based quantification techniques, TMT, IPTL, PRM, MALDI Imaging, SILAC, PTM analysis, DIA, cross-linking, and the up-to-date topics of software and 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, Quantitative Methods in Proteomics, Second Edition aims to provide comprehensive and competent overview in the important and still growing field of quantitative proteomics.

Quantitative Methods in Proteomics

Author : Katrin Marcus
Publisher : Humana Press
ISBN 13 : 9781617798849
Total Pages : 539 pages
Book Rating : 4.7/5 (988 download)

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Book Synopsis Quantitative Methods in Proteomics by : Katrin Marcus

Download or read book Quantitative Methods in Proteomics written by Katrin Marcus and published by Humana Press. This book was released on 2012-06-08 with total page 539 pages. Available in PDF, EPUB and Kindle. Book excerpt: Protein modifications and changes made to them, as well as the quantities of expressed proteins, can define the various functional stages of the cell. Accordingly, perturbations can lead to various diseases and disorders. As a result, it has become paramount to be able to detect and monitor post-translational modifications and to measure the abundance of proteins within the cell with extreme sensitivity. While protein identification is an almost routine requirement nowadays, reliable techniques for quantifying unmodified proteins (including those that escape detection under standard conditions, such as protein isoforms and membrane proteins) is not routine. Quantitative Methods in Proteomics gives a detailed survey of topics and methods on the principles underlying modern protein analysis, from statistical issues when planning proteomics experiments, to gel-based and mass spectrometry-based applications. The quantification of post-translational modifications is also addressed, followed by the “hot” topics of software and data analysis, as well as various overview chapters which provide a comprehensive overview of existing methods in quantitative proteomics. Written in the successful Methods in Molecular BiologyTM 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, Quantitative Methods in Proteomics serves as a comprehensive and competent overview of the important and still growing field of quantitative proteomics.

Systems Biology in Toxicology and Environmental Health

Author : Rebecca Fry
Publisher : Academic Press
ISBN 13 : 0128015683
Total Pages : 284 pages
Book Rating : 4.1/5 (28 download)

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Book Synopsis Systems Biology in Toxicology and Environmental Health by : Rebecca Fry

Download or read book Systems Biology in Toxicology and Environmental Health written by Rebecca Fry and published by Academic Press. This book was released on 2015-06-11 with total page 284 pages. Available in PDF, EPUB and Kindle. Book excerpt: Systems Biology in Toxicology and Environmental Health uses a systems biological perspective to detail the most recent findings that link environmental exposures to human disease, providing an overview of molecular pathways that are essential for cellular survival after exposure to environmental toxicants, recent findings on gene-environment interactions influencing environmental agent-induced diseases, and the development of computational methods to predict susceptibility to environmental agents. Introductory chapters on molecular and cellular biology, toxicology and computational biology are included as well as an assessment of systems-based tools used to evaluate environmental health risks. Further topics include research on environmental toxicants relevant to human health and disease, various high-throughput technologies and computational methods, along with descriptions of the biological pathways associated with disease and the developmental origins of disease as they relate to environmental contaminants. Systems Biology in Toxicology and Environmental Health is an essential reference for undergraduate students, graduate students, and researchers looking for an introduction in the use of systems biology approaches to assess environmental exposures and their impacts on human health. Provides the first reference of its kind, demonstrating the application of systems biology in environmental health and toxicology Includes introductions to the diverse fields of molecular and cellular biology, toxicology, and computational biology Presents a foundation that helps users understand the connections between the environment and health effects, and the biological mechanisms that link them

Biochemistry

Author : John T. Tansey
Publisher : John Wiley & Sons
ISBN 13 : 1119610559
Total Pages : 994 pages
Book Rating : 4.1/5 (196 download)

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Book Synopsis Biochemistry by : John T. Tansey

Download or read book Biochemistry written by John T. Tansey and published by John Wiley & Sons. This book was released on 2020-07-15 with total page 994 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biochemistry: An Integrative Approach with Expanded Topics is addressed to premed, biochemistry, and life science majors taking a two-semester biochemistry course. This version includes all 25 chapters, offering a holistic approach to learning biochemistry. An integrated, skill-focused approach to the study of biochemistry and metabolism Biochemistry integrates subjects of interest to undergraduates majoring in premed, biochemistry, life science, and beyond, while preserving a chemical perspective. Respected biochemistry educator John Tansey takes a unique approach to the subject matter, emphasizing problem solving and critical thinking over rote memorization. Key concepts such as metabolism, are introduced and then revisited and cross-referenced throughout the text to establish pattern recognition and help students commit their new knowledge to long-term memory. As part of WileyPLUS, Biochemistry includes access to video walkthroughs of worked problems, interactive elements, and expanded end-of-chapter problems with a wide range of subject matter and difficulty. Students will have access to both qualitative and quantitative worked problems, and videos model the biochemical reasoning students will need to master. This approach helps students learn to analyze data and make critical assessments of experiments—key skills for success across scientific disciplines. Introduces students in scientific majors to the basics of biochemistry and metabolism Integrates and synthesizes topics throughout the text, allowing students to learn through repetition and pattern recognition Emphasizes problem solving and reasoning skills essential to life sciences, including data analysis and research assessment Provides access to video walkthroughs of worked problems, interactive features, and additional study material through WileyPLUS This volume covers DNA, RNA, gene regulation, synthetic proteins, omics, plant biochemistry, and more. With this text, students studying a range of disciplines are empowered to develop a lasting foundation in biochemistry and metabolism that will serve them as they advance through their careers.

Encyclopedia of Bioinformatics and Computational Biology

Author :
Publisher : Elsevier
ISBN 13 : 0128114320
Total Pages : 3421 pages
Book Rating : 4.1/5 (281 download)

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Book Synopsis Encyclopedia of Bioinformatics and Computational Biology by :

Download or read book Encyclopedia of Bioinformatics and Computational Biology written by and published by Elsevier. This book was released on 2018-08-21 with total page 3421 pages. Available in PDF, EPUB and Kindle. Book excerpt: Encyclopedia of Bioinformatics and Computational Biology: ABC of Bioinformatics, Three Volume Set combines elements of computer science, information technology, mathematics, statistics and biotechnology, providing the methodology and in silico solutions to mine biological data and processes. The book covers Theory, Topics and Applications, with a special focus on Integrative –omics and Systems Biology. The theoretical, methodological underpinnings of BCB, including phylogeny are covered, as are more current areas of focus, such as translational bioinformatics, cheminformatics, and environmental informatics. Finally, Applications provide guidance for commonly asked questions. This major reference work spans basic and cutting-edge methodologies authored by leaders in the field, providing an invaluable resource for students, scientists, professionals in research institutes, and a broad swath of researchers in biotechnology and the biomedical and pharmaceutical industries. Brings together information from computer science, information technology, mathematics, statistics and biotechnology Written and reviewed by leading experts in the field, providing a unique and authoritative resource Focuses on the main theoretical and methodological concepts before expanding on specific topics and applications Includes interactive images, multimedia tools and crosslinking to further resources and databases

HT-MS Tools for Data Quantification

Author : Joanne Ava
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.8/5 (689 download)

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Book Synopsis HT-MS Tools for Data Quantification by : Joanne Ava

Download or read book HT-MS Tools for Data Quantification written by Joanne Ava and published by . This book was released on 2023-10-27 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Computational Medicine

Author : Zlatko Trajanoski
Publisher : Springer Science & Business Media
ISBN 13 : 3709109477
Total Pages : 207 pages
Book Rating : 4.7/5 (91 download)

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Book Synopsis Computational Medicine by : Zlatko Trajanoski

Download or read book Computational Medicine written by Zlatko Trajanoski and published by Springer Science & Business Media. This book was released on 2012-09-18 with total page 207 pages. Available in PDF, EPUB and Kindle. Book excerpt: Computational methodologies and modeling play a growing role for investigating mechanisms, and for the diagnosis and therapy of human diseases. This progress gave rise to computational medicine, an interdisciplinary field at the interface of computer science and medicine. The main focus of computational medicine lies in the development of data analysis methods and mathematical modeling as well as computational simulation techniques specifically addressing medical problems. In this book, we present a number of computational medicine topics at several scales: from molecules to cells, organs, and organisms. At the molecular level, tools for the analysis of genome variations as well as cloud computing resources for medical genetics are reviewed. Then, an analysis of gene expression data and the application to the characterization of microbial communities are highlighted. At the protein level, two types of analyses for mass spectrometry data are reviewed: labeled quantitative proteomics and lipidomics, followed by protein sequence analysis and a 3D structure and drug design chapter. Finally, three chapters on clinical applications focus on the integration of biomolecular and clinical data for cancer research, biomarker discovery, and network-based methods for computational diagnostics.

Statistical Analysis of Proteomic Data

Author : Thomas Burger
Publisher : Springer Nature
ISBN 13 : 1071619675
Total Pages : 398 pages
Book Rating : 4.0/5 (716 download)

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Book Synopsis Statistical Analysis of Proteomic Data by : Thomas Burger

Download or read book Statistical Analysis of Proteomic Data written by Thomas Burger and published by Springer Nature. This book was released on 2022-10-29 with total page 398 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book explores the most important processing steps of proteomics data analysis and presents practical guidelines, as well as software tools, that are both user-friendly and state-of-the-art in chemo- and biostatistics. Beginning with methods to control the false discovery rate (FDR), the volume continues with chapters devoted to software suites for constructing quantitation data tables, missing value related issues, differential analysis software, and more. Written for the highly successful Methods in Molecular Biology series, chapters include the kind of detail and implementation advice that leads to successful results. Authoritative and practical, Statistical Analysis of Proteomic Data: Methods and Tools serves as an ideal guide for proteomics researchers looking to extract the best of their data with state-of-the art tools while also deepening their understanding of data analysis.

Biological Knowledge Discovery Handbook

Author : Mourad Elloumi
Publisher : John Wiley & Sons
ISBN 13 : 1118853725
Total Pages : 1192 pages
Book Rating : 4.1/5 (188 download)

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Book Synopsis Biological Knowledge Discovery Handbook by : Mourad Elloumi

Download or read book Biological Knowledge Discovery Handbook written by Mourad Elloumi and published by John Wiley & Sons. This book was released on 2015-02-04 with total page 1192 pages. Available in PDF, EPUB and Kindle. Book excerpt: The first comprehensive overview of preprocessing, mining,and postprocessing of biological data Molecular biology is undergoing exponential growth in both thevolume and complexity of biological data—and knowledgediscovery offers the capacity to automate complex search and dataanalysis tasks. This book presents a vast overview of the mostrecent developments on techniques and approaches in the field ofbiological knowledge discovery and data mining (KDD)—providingin-depth fundamental and technical field information on the mostimportant topics encountered. Written by top experts, Biological Knowledge DiscoveryHandbook: Preprocessing, Mining, and Postprocessing of BiologicalData covers the three main phases of knowledge discovery (datapreprocessing, data processing—also known as datamining—and data postprocessing) and analyzes both verificationsystems and discovery systems. BIOLOGICAL DATA PREPROCESSING Part A: Biological Data Management Part B: Biological Data Modeling Part C: Biological Feature Extraction Part D Biological Feature Selection BIOLOGICAL DATA MINING Part E: Regression Analysis of Biological Data Part F Biological Data Clustering Part G: Biological Data Classification Part H: Association Rules Learning from Biological Data Part I: Text Mining and Application to Biological Data Part J: High-Performance Computing for Biological DataMining Combining sound theory with practical applications in molecularbiology, Biological Knowledge Discovery Handbook is idealfor courses in bioinformatics and biological KDD as well as forpractitioners and professional researchers in computer science,life science, and mathematics.

Proteome Informatics

Author : Conrad Bessant
Publisher : Royal Society of Chemistry
ISBN 13 : 1782626735
Total Pages : 412 pages
Book Rating : 4.7/5 (826 download)

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Book Synopsis Proteome Informatics by : Conrad Bessant

Download or read book Proteome Informatics written by Conrad Bessant and published by Royal Society of Chemistry. This book was released on 2016-11-15 with total page 412 pages. Available in PDF, EPUB and Kindle. Book excerpt: The field of proteomics has developed rapidly over the past decade nurturing the need for a detailed introduction to the various informatics topics that underpin the main liquid chromatography tandem mass spectrometry (LC-MS/MS) protocols used for protein identification and quantitation. Proteins are a key component of any biological system, and monitoring proteins using LC-MS/MS proteomics is becoming commonplace in a wide range of biological research areas. However, many researchers treat proteomics software tools as a black box, drawing conclusions from the output of such tools without considering the nuances and limitations of the algorithms on which such software is based. This book seeks to address this situation by bringing together world experts to provide clear explanations of the key algorithms, workflows and analysis frameworks, so that users of proteomics data can be confident that they are using appropriate tools in suitable ways.