Peptides And Peptide Based Biomaterials And Their Biomedical Applications

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Peptides and Peptide-based Biomaterials and their Biomedical Applications

Author : Anwar Sunna
Publisher : Springer
ISBN 13 : 3319660950
Total Pages : 309 pages
Book Rating : 4.3/5 (196 download)

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Book Synopsis Peptides and Peptide-based Biomaterials and their Biomedical Applications by : Anwar Sunna

Download or read book Peptides and Peptide-based Biomaterials and their Biomedical Applications written by Anwar Sunna and published by Springer. This book was released on 2017-10-26 with total page 309 pages. Available in PDF, EPUB and Kindle. Book excerpt: Solid-binding peptides have been used increasingly as molecular building blocks in nanobiotechnology as they can direct the assembly and functionalisation of a diverse range of materials and have the ability to regulate the synthesis of nanoparticles and complex nanostructures. Nanostructured materials such as β-sheet fibril-forming peptides and α-helical coiled coil systems have displayed many useful properties including stimulus-responsiveness, modularity and multi-functionality, providing potential technological applications in tissue engineering, antimicrobials, drug delivery and nanoscale electronics. The current situation with respect to self-assembling peptides and bioactive matrices for regenerative medicine are reviewed, as well as peptide-target modeling and an examination of future prospects for peptides in these areas.

Peptide-based Biomaterials

Author : Mustafa O. Guler
Publisher : Royal Society of Chemistry
ISBN 13 : 1839161159
Total Pages : 413 pages
Book Rating : 4.8/5 (391 download)

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Book Synopsis Peptide-based Biomaterials by : Mustafa O. Guler

Download or read book Peptide-based Biomaterials written by Mustafa O. Guler and published by Royal Society of Chemistry. This book was released on 2020-11-18 with total page 413 pages. Available in PDF, EPUB and Kindle. Book excerpt: Research and new tools in biomaterials development by using peptides are currently growing, as more functional and versatile building blocks are used to design a host of functional biomaterials via chemical modifications for health care applications. It is a field that is attracting researchers from across soft matter science, molecular engineering and biomaterials science. Covering the fundamental concepts of self-assembly, design and synthesis of peptides, this book will provide a solid introduction to the field for those interested in developing functional biomaterials by using peptide derivatives. The bioactive nature of the peptides and their physical properties are discussed in various applications in biomedicine. This book will help researchers and students working in biomaterials and biomedicine fields and help their understanding of modulating biological processes for disease diagnosis and treatments.

Peptides and Proteins as Biomaterials for Tissue Regeneration and Repair

Author : Mario Barbosa
Publisher : Woodhead Publishing
ISBN 13 : 008100852X
Total Pages : 392 pages
Book Rating : 4.0/5 (81 download)

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Book Synopsis Peptides and Proteins as Biomaterials for Tissue Regeneration and Repair by : Mario Barbosa

Download or read book Peptides and Proteins as Biomaterials for Tissue Regeneration and Repair written by Mario Barbosa and published by Woodhead Publishing. This book was released on 2017-09-25 with total page 392 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peptides and Proteins as Biomaterials for Tissue Regeneration and Repair highlights the various important considerations that go into biomaterial development, both in terms of fundamentals and applications. After covering a general introduction to protein and cell interactions with biomaterials, the book discusses proteins in biomaterials that mimic the extracellular matrix (ECM). The properties, fabrication and application of peptide biomaterials and protein-based biomaterials are discussed in addition to in vivo and in vitro studies. This book is a valuable resource for researchers, scientists and advanced students interested in biomaterials science, chemistry, molecular biology and nanotechnology. Presents an all-inclusive and authoritative coverage of the important role which protein and peptides play as biomaterials for tissue regeneration Explores protein and peptides from the fundamentals, to processing and applications Written by an international group of leading biomaterials researchers

Peptide Applications in Biomedicine, Biotechnology and Bioengineering

Author : Sotirios Koutsopoulos
Publisher : Woodhead Publishing
ISBN 13 : 0081007426
Total Pages : 655 pages
Book Rating : 4.0/5 (81 download)

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Book Synopsis Peptide Applications in Biomedicine, Biotechnology and Bioengineering by : Sotirios Koutsopoulos

Download or read book Peptide Applications in Biomedicine, Biotechnology and Bioengineering written by Sotirios Koutsopoulos and published by Woodhead Publishing. This book was released on 2017-11-16 with total page 655 pages. Available in PDF, EPUB and Kindle. Book excerpt: Peptide Applications in Biomedicine, Biotechnology and Bioengineering summarizes the current knowledge on peptide applications in biomedicine, biotechnology and bioengineering. After a general introduction to peptides, the book addresses the many applications of peptides in biomedicine and medical technology. Next, the text focuses on peptide applications in biotechnology and bioengineering and reviews of peptide applications in nanotechnology. This book is a valuable resource for biomaterial scientists, polymer scientists, bioengineers, mechanical engineers, synthetic chemists, medical doctors and biologists. Presents a self-contained work for the field of biomedical peptides Summarizes the current knowledge on peptides in biomedicine, biotechnology and bioengineering Covers current and potential applications of biomedical peptides

Peptide Bionanomaterials

Author : Mohamed A. Elsawy
Publisher : Springer Nature
ISBN 13 : 3031293606
Total Pages : 556 pages
Book Rating : 4.0/5 (312 download)

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Book Synopsis Peptide Bionanomaterials by : Mohamed A. Elsawy

Download or read book Peptide Bionanomaterials written by Mohamed A. Elsawy and published by Springer Nature. This book was released on 2023-08-28 with total page 556 pages. Available in PDF, EPUB and Kindle. Book excerpt: Molecular self-assembly has been exploited by nature for developing the higher functional macromolecular structures of both the genome and proteome. Inspired by nature, there has been a surge of research, in the last two decades, for the molecular engineering of peptide-based self-assembling nanostructures, adopting the bottom-up design approach. This book gives the reader an overview on the design rules for de novo self-assembling peptide and reviews the diverse range of bioinspired peptide nanostructures such as β-sheet and β-hairpin, α-helical and coiled coil, self-assembling short peptides and peptidomimetics, collagen-based and elastin-like peptides, silk peptides, peptide amphiphiles, peptides co-polymers and others. The book also covers the wide variety of responsive and functional biomaterials that have been innovated based on those nanostructures for various applications ranging from tissue engineering, therapeutics and drug delivery to antimicrobial nanomaterials and biosensors. Finally, the book also discusses the peptide bionanomaterials global market and the future of the emerging industry. Chapter “Characterization of Peptide-Based Nanomaterials” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Design and Characterization of Peptide-based Biomaterials

Author : Sivakumar Ramachandran
Publisher :
ISBN 13 : 9781109918731
Total Pages : 94 pages
Book Rating : 4.9/5 (187 download)

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Book Synopsis Design and Characterization of Peptide-based Biomaterials by : Sivakumar Ramachandran

Download or read book Design and Characterization of Peptide-based Biomaterials written by Sivakumar Ramachandran and published by . This book was released on 2007 with total page 94 pages. Available in PDF, EPUB and Kindle. Book excerpt: The main objective of this dissertation is to lay out the design principles involved in assembling stimuli-sensitive, peptide-based biomaterials that has potential for various biomedical applications like tissue engineering and drug delivery. Supramolecular systems, which enable one to assemble novel materials from molecular level, have fascinated researchers in many disciplines. Inspired by such systems, a set of mutually complementary, self-repulsive oligopeptide modules (with alternating polar-apolar amino acid sequence) were designed to gain better control and wide range of tunability over the assembling process. These peptide modules (at 0.25 wt% in aqueous buffer) assembled into a hydrogel network with change in pH/ionic strength (self-assembly) and upon mixing the two mutually-complementary peptide modules (co-assembly). Mixing induced hydrogels are particularly attractive as they can be easily assembled by simple mixing of peptide solutions prior to application. Another advantage of mixing-induced gelation is that it preserves the pH and ionic strength of the original peptide solutions. Circular dichroism spectroscopy of individual decapeptide solutions revealed their random coil conformation. Transmission electron microscopy images showed the nanofibrillar network structure of the hydrogel. Dynamic rheological characterization revealed its high elasticity and shear-thinning nature. Furthermore, the co-assembled hydrogel was capable of rapid recoveries from repeated shear-induced breakdowns, a property desirable for designing injectable biomaterials. A systematic variation of the neutral amino acids in the sequence revealed some of the design principles for this class of biomaterials. First, viscoelastic properties of the hydrogels can be tuned through adjusting the hydrophobicity of the neutral amino acids. Second, the beta-sheet propensity of the neutral amino acid residue in the peptides is critical for hydrogelation. The compatibility of these hydrogels with entrapped biomolecules (molecular biocompatibility) was confirmed using high-resolution, 1H-15N heteronuclear NMR spectroscopy. Sticky-ends in the peptide fibers did not provide any advantages over blunt-ends in terms of biomaterial property, suggesting that peptides might be aligned perpendicular to the fiber axis. Once the design principles of these peptide-based biomaterials are laid out, the next step would be to incorporate biological functionalities like cell-adhesive motifs and enzyme recognition sequences at strategic positions of these biomaterials for specific tissue engineering or drug delivery applications.

Engineering a Peptide-based Tool for Biomedical Applications

Author : Seren Hamsici
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (139 download)

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Book Synopsis Engineering a Peptide-based Tool for Biomedical Applications by : Seren Hamsici

Download or read book Engineering a Peptide-based Tool for Biomedical Applications written by Seren Hamsici and published by . This book was released on 2022 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:

Peptide-Based Materials

Author : Timothy Deming
Publisher : Springer Science & Business Media
ISBN 13 : 3642271391
Total Pages : 184 pages
Book Rating : 4.6/5 (422 download)

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Book Synopsis Peptide-Based Materials by : Timothy Deming

Download or read book Peptide-Based Materials written by Timothy Deming and published by Springer Science & Business Media. This book was released on 2012-01-10 with total page 184 pages. Available in PDF, EPUB and Kindle. Book excerpt: Synthesis of Polypeptides by Ring-Opening Polymerization of α-Amino Acid N-Carboxyanhydrides, by Jianjun Cheng and Timothy J. Deming.- Peptide Synthesis and Self-Assembly, by S. Maude, L. R. Tai, R. P. W. Davies, B. Liu, S. A. Harris, P. J. Kocienski and A. Aggeli.- Elastomeric Polypeptides, by Mark B. van Eldijk, Christopher L. McGann, Kristi L. Kiick andJan C. M. van Hest.- Self-Assembled Polypeptide and Polypeptide Hybrid Vesicles: From Synthesis to Application, by Uh-Joo Choe, Victor Z. Sun, James-Kevin Y. Tan and Daniel T. Kamei.- Peptide-Based and Polypeptide-Based Hydrogels for Drug Delivery and Tissue Engineering, by Aysegul Altunbas and Darrin J. Pochan.-

Self-Assembled Peptide Nanostructures

Author : Jaime Castillo
Publisher : CRC Press
ISBN 13 : 9814316946
Total Pages : 326 pages
Book Rating : 4.8/5 (143 download)

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Book Synopsis Self-Assembled Peptide Nanostructures by : Jaime Castillo

Download or read book Self-Assembled Peptide Nanostructures written by Jaime Castillo and published by CRC Press. This book was released on 2012-11-21 with total page 326 pages. Available in PDF, EPUB and Kindle. Book excerpt: The self-organization of bionanostructures into well-defined functional machineries found in nature has been a priceless source of ideas for researchers. The molecules of life, proteins, DNA, RNA, etc., as well as the structures and forms that these molecules assume serve as rich sources of ideas for scientists or engineers who are interested in developing bio-inspired materials for innovations in biomedical fields. In nature, molecular self-assembly is a process by which complex three-dimensional structures with well-defined functions are constructed, starting from simple building blocks such as proteins and peptides. This book introduces readers to the theory and mechanisms of peptide self-assembly processes. The authors present the more common peptide self-assembled building blocks and discuss how researchers from different fields can apply self-assembling principles to bionanotechnology applications. The advantages and challenges are mentioned together with examples that reflect the state of the art of the use of self-assembled peptide building blocks in nanotechnology.

Folded Synthetic Peptides for Biomedical Applications

Author : Alessandro Contini
Publisher : Frontiers Media SA
ISBN 13 : 2889630455
Total Pages : 145 pages
Book Rating : 4.8/5 (896 download)

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Book Synopsis Folded Synthetic Peptides for Biomedical Applications by : Alessandro Contini

Download or read book Folded Synthetic Peptides for Biomedical Applications written by Alessandro Contini and published by Frontiers Media SA. This book was released on 2019-09-26 with total page 145 pages. Available in PDF, EPUB and Kindle. Book excerpt: Folded peptides - and peptide motifs within proteins - are abundant in living organisms, where they are essential for the biological activities of the peptides and proteins. During the past decades, much research has been dedicated to understanding the rules that govern peptide folding. Simultaneously, a range of strategies have been established for the conformational stabilization of bioactive peptides, as well as for the de novo design of peptides with defined secondary structures. These methods are either based on the chemical modification of the peptide backbone, such as cyclization and stapled peptides, or on the use of a range of non-proteinogenic amino acids that, in a defined sequential arrangement, induce secondary structures peptides. Such building blocks include D- and other non-proteinogenic amino acids, as well as beta- and gamma-amino acids. This Research Topic comprises a collection of papers by an international group of 77 scientists with a background in synthetic, analytical, computational and medicinal chemistry, as well as in biochemistry and pharmacology. Their research is presented here in a total of 11 papers (8 original research reports and 3 reviews), covering diverse aspects of folded synthetic peptides. These studies include the preparation and characterization of new peptide monomers with interesting folding properties, the synthesis and conformational analysis of non-natural peptides, as well as the use of folded peptidomimetics as molecular switches. Additionally, a range of biomedical applications, such as antimicrobial, anti-inflammatory, antiangiogenic and immune-stimulating activities, are also reported. We hope this eBook will be a source of inspiration and knowledge for scientist in various disciplines related to folded peptides and their many applications, as well as for those who want to learn more about this fascinating field of research.

Biopolymer Science for Proteins and Peptides

Author : Keiji Numata
Publisher : Elsevier
ISBN 13 : 0128205563
Total Pages : 310 pages
Book Rating : 4.1/5 (282 download)

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Book Synopsis Biopolymer Science for Proteins and Peptides by : Keiji Numata

Download or read book Biopolymer Science for Proteins and Peptides written by Keiji Numata and published by Elsevier. This book was released on 2021-08-14 with total page 310 pages. Available in PDF, EPUB and Kindle. Book excerpt: Biopolymer Science for Proteins and Peptides introduces all aspects of natural polymers based on structural proteins and peptides, presenting synthesis, structure, properties, proteins, materials design, and applications. The book begins by presenting the core concepts of polypeptide and protein materials, before discussing synthesis and structure in detail. The next part of the book describes physical properties, biological properties, and issues surrounding stability. Subsequent chapters offer in-depth coverage of both natural and structural protein sources, including collagen, silk, elastin, resilin, keratin, foot protein, and reflectin, and the materials that can be designed from them, such as films, fibers, textiles, microparticles, sponges and scaffolds, nanomaterials, blends, and composites. These materials are also analyzed against the available synthetic polymers. Finally, the text explores current applications and potential future developments. This is an essential resource for researchers and advanced students across a range of disciplines, including biopolymers, structural proteins, polymer science, materials science, biomaterials, biology, biotechnology, chemistry, engineering, and pharmaceutical science. In an industry setting, this is of great interest to scientists and R&D professionals working in industries with an interest in bio-based polymers for advanced applications. Explains how biopolymers from structural proteins and peptides can be developed into materials, such as films, fibers, textiles, microparticles, sponges and scaffolds, nanomaterials, blends, and polymer composites Provides the reader a solid understanding of the structure, synthesis, and properties Guides the reader from sources, including collagen, silk, elastin, resilin, keratin, and reflectin, to material design and cutting-edge applications

Biomaterials from Nature for Advanced Devices and Therapies

Author : Nuno M. Neves
Publisher : John Wiley & Sons
ISBN 13 : 1118478053
Total Pages : 724 pages
Book Rating : 4.1/5 (184 download)

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Book Synopsis Biomaterials from Nature for Advanced Devices and Therapies by : Nuno M. Neves

Download or read book Biomaterials from Nature for Advanced Devices and Therapies written by Nuno M. Neves and published by John Wiley & Sons. This book was released on 2016-10-24 with total page 724 pages. Available in PDF, EPUB and Kindle. Book excerpt: In-depth information on natural biomaterials and their applications for translational medicine! Undiluted expertise: edited by world-leading experts with contributions from top-notch international scientists, collating experience and cutting-edge knowledge on natural biomaterials from all over the world A must-have on the shelf in every biomaterials lab: graduate and PhD students beginning their career in biomaterials science and experienced researchers and practitioners alike will turn to this comprehensive reference in their daily work Link to clinical practice: chapters on translational research make readers aware of what needs to be considered when a biomaterial leaves the lab to be routinely used

Artificial Protein and Peptide Nanofibers

Author : Gang Wei
Publisher : Woodhead Publishing
ISBN 13 : 0081028512
Total Pages : 504 pages
Book Rating : 4.0/5 (81 download)

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Book Synopsis Artificial Protein and Peptide Nanofibers by : Gang Wei

Download or read book Artificial Protein and Peptide Nanofibers written by Gang Wei and published by Woodhead Publishing. This book was released on 2020-07-28 with total page 504 pages. Available in PDF, EPUB and Kindle. Book excerpt: Artificial Protein and Peptide Nanofibers: Design, Fabrication, Characterization, and Applications provides comprehensive knowledge of the preparation, modification and applications of protein and peptide nanofibers. The book reviews the synthesis and strategies necessary to create protein and peptide nanofibers, such as self-assembly (including supramolecular assembly), electrospinning, template synthesis, and enzymatic synthesis. Then, the key chemical modification and molecular design methods are highlighted that can be utilized to improve the bio-functions of these synthetic fibers. Finally, fabrication methods for key applications, such as sensing, drug delivery, imaging, tissue engineering and electronic devices are reviewed. This book will be an ideal resource for those working in materials science, polymer science, chemical engineering, nanotechnology and biomedicine. Reviews key chemical modification and molecular design methods to improve the bio-functions of synthetic peptide and protein nanofibers Discusses the most important synthesis strategies, including supramolecular assembly, electrospinning, template synthesis and enzymatic synthesis Provides information on fabrication of nanofibers for key applications such as sensing, imaging, drug delivery and tissue engineering

Simulation of Protein and Peptide-Based Biomaterials

Author :
Publisher :
ISBN 13 :
Total Pages : 0 pages
Book Rating : 4.:/5 (946 download)

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Book Synopsis Simulation of Protein and Peptide-Based Biomaterials by :

Download or read book Simulation of Protein and Peptide-Based Biomaterials written by and published by . This book was released on 2002 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: The overall goal of the proposed research is to pursue realistic molecular modeling studies of the stability, dynamics, structure, function, and folding of proteins and protein-based biomaterials in solution. Simulations of elastin-based polymers will investigate the driving forces and mechanism for the unusual temperature-dependent conformational changes, knowledge that will assist in redesign of this system for specific applications and the design of other such molecules. Studies of the stabilities of cyclic peptides and their multimers (nanotubes) to both environment and peptide substitutions will be addressed, as well as their mechanism of transport, selectivity, and gating. Methods are being developed for identifying and optimizing small, robust scaffolds for use as novel receptors, or biosensors. Functional optimization (typically binding) will also be explored.

Engineering New Self-assembling Biomaterials Based on Beta-sheet-forming Peptides

Author : Joel Henderson Collier
Publisher :
ISBN 13 :
Total Pages : pages
Book Rating : 4.:/5 (549 download)

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Book Synopsis Engineering New Self-assembling Biomaterials Based on Beta-sheet-forming Peptides by : Joel Henderson Collier

Download or read book Engineering New Self-assembling Biomaterials Based on Beta-sheet-forming Peptides written by Joel Henderson Collier and published by . This book was released on 2003 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: This thesis deals with the development of novel strategies for self-assembling biomaterials based on short beta-sheet fibril-forming peptides. These materials are potentially useful for a variety of biomedical applications, including tissue engineering and controlled drug release, and several aspects of these materials have been explored in this thesis in an effort to improve their clinical utility. First, a strategy for rapidly and uniformly triggering peptide self-assembly was developed. Stimulus-sensitive liposomes were utilized to sequester salts and release them in response to stimuli such as warming to body temperature or exposure to near infra-red light. Triggered salt release in turn initiated rapid self-assembly of extravesicular peptide and the transformation of the peptide/liposome suspension from a solution to a hydrogel. We show that peptide self-assembly can be rapidly, uniformly, and specifically triggered, with the storage modulus increasing three orders of magnitude upon triggering. This rapid triggering strategy is potentially useful for injectable biomaterial applications. In the second research thrust, a self-assembling peptide was developed that also possessed enzymatic activity for the cross-linking enzyme tissue transglutaminase. We then utilized tissue transglutaminase to conjugate cell-interactive peptides to the self-assembled structure and investigated cell attachment to these functionalized scaffolds. Such a strategy is potentially useful for designing biospecific self-assembling scaffolds for tissue engineering or wound healing. In the final research focus, to modulate the nanostructure of the self-assembled fibrillar structures, we developed a series of self-assembling polyethylene glycol-conjugated peptides. We found that polyethylene glycol conjugation significantly altered fibril morphology, including width, length, and degree of lateral aggregation. As previous beta-sheet fibrillar self-assembled materials have shown particular intransigence to such modification of the fibrillar structure, this strategy may be a useful route for tailoring the physical properties of these materials. As a whole, the strategies developed in this thesis address previous shortcomings of beta-sheet peptide-based self-assembling materials in an effort to bring them closer to clinical application.

De Novo Peptide Design

Author : Vibin Ramakrishnan
Publisher : Academic Press
ISBN 13 : 0323985432
Total Pages : 297 pages
Book Rating : 4.3/5 (239 download)

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Book Synopsis De Novo Peptide Design by : Vibin Ramakrishnan

Download or read book De Novo Peptide Design written by Vibin Ramakrishnan and published by Academic Press. This book was released on 2022-09-17 with total page 297 pages. Available in PDF, EPUB and Kindle. Book excerpt: De novo Peptide Design: Principles and Applications presents the latest developments in the fields of therapeutic peptides and bio-nanotechnology. The title focuses on the design of peptides, particularly how peptides may be tailored to specific functions. It includes computational and experimental protocols to assist in the design of peptides. Sections cover the basics of protein and peptide structure, modeling and simulation, solid phase peptide synthesis, peptide-based antibiotics, drug delivery, peptide nanomaterials, aromatic interactions directing nano-assembly, protein/peptide aggregation, therapeutic interventions against protein/peptide aggregation diseases, peptide based hydrogels, computational tools and algorithms for peptide design, and experimental protocols in peptide chemistry. In addition, the book covers key aspects in peptide design, providing a solution for researchers working within the 'peptidic universe' to create new therapeutic agents. Gives comprehensive coverage, including peptide design, modeling, synthesis and applications Presents emerging topics in the design of peptide-based therapeutics Details the latest developments in the fields of therapeutic peptides and bio-nanotechnology Considers peptide design and the tailoring of peptides to specific functions Offers computational tools and algorithms for peptide design and experimental protocols for peptide chemistry

Cellulose-Based Superabsorbent Hydrogels

Author : Md. Ibrahim H. Mondal
Publisher : Springer
ISBN 13 : 9783319778297
Total Pages : 0 pages
Book Rating : 4.7/5 (782 download)

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Book Synopsis Cellulose-Based Superabsorbent Hydrogels by : Md. Ibrahim H. Mondal

Download or read book Cellulose-Based Superabsorbent Hydrogels written by Md. Ibrahim H. Mondal and published by Springer. This book was released on 2019-02-22 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt: With the prospect of revolutionizing specific technologies, this book highlights the most exciting and impactful current research in the fields of cellulose-based superabsorbent hydrogels with their smart applications. The book assembles the newest synthetic routes, characterization methods, and applications in the emergent area. Leading experts in the field have contributed chapters representative of their most recent research results, shedding light on the enormous potential of this field and thoroughly presenting cellulose-based hydrogel functioning materials. The book is intended for the polymer chemists, academic and industrial scientists and engineers, pharmaceutical and biomedical scientists and agricultural engineers engaged in research and development on absorbency, absorbent products and superabsorbent hydrogels. It can also be supportive for undergraduate and graduate students.