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Released this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\\n\\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
\\n"}]',published:!0,mainMedia:{caption:"Highly Cited",originalUrl:"/media/original/117"}},components:[{type:"htmlEditorComponent",content:'IntechOpen is proud to announce that 191 of our authors have made the Clarivate™ Highly Cited Researchers List for 2020, ranking them among the top 1% most-cited.
\n\nThroughout the years, the list has named a total of 261 IntechOpen authors as Highly Cited. Of those researchers, 69 have been featured on the list multiple times.
\n\n\n\nReleased this past November, the list is based on data collected from the Web of Science and highlights some of the world’s most influential scientific minds by naming the researchers whose publications over the previous decade have included a high number of Highly Cited Papers placing them among the top 1% most-cited.
\n\nWe wish to congratulate all of the researchers named and especially our authors on this amazing accomplishment! We are happy and proud to share in their success!
Note: Edited in March 2021
\n'}],latestNews:[{slug:"webinar-introduction-to-open-science-wednesday-18-may-1-pm-cest-20220518",title:"Webinar: Introduction to Open Science | Wednesday 18 May, 1 PM CEST"},{slug:"step-in-the-right-direction-intechopen-launches-a-portfolio-of-open-science-journals-20220414",title:"Step in the Right Direction: IntechOpen Launches a Portfolio of Open Science Journals"},{slug:"let-s-meet-at-london-book-fair-5-7-april-2022-olympia-london-20220321",title:"Let’s meet at London Book Fair, 5-7 April 2022, Olympia London"},{slug:"50-books-published-as-part-of-intechopen-and-knowledge-unlatched-ku-collaboration-20220316",title:"50 Books published as part of IntechOpen and Knowledge Unlatched (KU) Collaboration"},{slug:"intechopen-joins-the-united-nations-sustainable-development-goals-publishers-compact-20221702",title:"IntechOpen joins the United Nations Sustainable Development Goals Publishers Compact"},{slug:"intechopen-signs-exclusive-representation-agreement-with-lsr-libros-servicios-y-representaciones-s-a-de-c-v-20211123",title:"IntechOpen Signs Exclusive Representation Agreement with LSR Libros Servicios y Representaciones S.A. de C.V"},{slug:"intechopen-expands-partnership-with-research4life-20211110",title:"IntechOpen Expands Partnership with Research4Life"},{slug:"introducing-intechopen-book-series-a-new-publishing-format-for-oa-books-20210915",title:"Introducing IntechOpen Book Series - A New Publishing Format for OA Books"}]},book:{item:{type:"book",id:"5874",leadTitle:null,fullTitle:"Structural Health Monitoring - Measurement Methods and Practical Applications",title:"Structural Health Monitoring",subtitle:"Measurement Methods and Practical Applications",reviewType:"peer-reviewed",abstract:"Structural health monitoring (SHM) is a new engineering field with a growing tendency, based on technology development focused on data acquisition and analysis, to prevent possible damage in man-made structures and land's natural faults. The data are obtained from sensors and monitoring systems that allow detecting damages on structures, space vehicles, and land natural faults, to model their behavior under adverse scenarios, in order to search the detection of anomalies. Currently, there are many SHM systems with sensors based on different technologies like optical fiber, video cameras, optical scanners, wireless networks, and piezoelectric transducers, among others. In this context, the present book includes selected chapters with theoretical models and applications, to preserve infrastructure and prevent loss of human lives.",isbn:"978-953-51-3254-7",printIsbn:"978-953-51-3253-0",pdfIsbn:"978-953-51-4787-9",doi:"10.5772/65818",price:119,priceEur:129,priceUsd:155,slug:"structural-health-monitoring-measurement-methods-and-practical-applications",numberOfPages:140,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"72f61895d84252f6f5b92b7625741743",bookSignature:"Moises Rivas-Lopez, Wendy Flores Fuentes and Oleg Sergiyenko",publishedDate:"June 21st 2017",coverURL:"https://cdn.intechopen.com/books/images_new/5874.jpg",numberOfDownloads:8150,numberOfWosCitations:7,numberOfCrossrefCitations:4,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:11,numberOfDimensionsCitationsByBook:0,hasAltmetrics:0,numberOfTotalCitations:22,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 17th 2016",dateEndSecondStepPublish:"November 21st 2016",dateEndThirdStepPublish:"February 3rd 2017",dateEndFourthStepPublish:"May 4th 2017",dateEndFifthStepPublish:"July 3rd 2017",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6,7",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"178178",title:"Dr.",name:"Moises",middleName:null,surname:"Rivas-Lopez",slug:"moises-rivas-lopez",fullName:"Moises Rivas-Lopez",profilePictureURL:"https://mts.intechopen.com/storage/users/178178/images/4675_n.jpg",biography:"Dr. Rivas was born in 1960. He received the BS and MS degrees in Autonomous University of Baja California, Mexico, in 1985 and 1991, respectively, and the PhD degree in Science, Applied Physics, in the same University, in 2010.\nHe has written 5 book chapters and 35 Journal and Proceedings Conference papers in optoelectronics and control applications. Also, he has presented different works in several International Congresses of IEEE, ICROS, SICE, in America and Europe.\nDr. Rivas was Dean of Engineering Institute of Autonomous University Baja California (1997–2005) and Rector of Polytechnic University of Baja California (2006–2010). He is a member of National Researcher System and now is the head of Physics Engineering Department, of Engineering Institute of UABC, Mexico.",institutionString:null,position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Autonomous University of Baja California",institutionURL:null,country:{name:"Mexico"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:{id:"154068",title:"MSc.",name:"Wendy",middleName:null,surname:"Flores F.",slug:"wendy-flores-f.",fullName:"Wendy Flores F.",profilePictureURL:"https://mts.intechopen.com/storage/users/154068/images/4929_n.jpg",biography:"Dr. Flores-Fuentes was born in Baja California, Mexico on January, 1978. She received the bachelor\\'s degree in Electronic Engineering from the Autonomous University of Baja California in 2001, the master’s in Engineering degree from Technological Institute of Mexicali in 2006, and the PhD. degree in Science, Applied Physics, from Autonomous University of Baja California in June 2014. Until now she is the author of 4 journal articles, 2 book chapters, and 13 proceedings articles. Recently she organized and participated as Chair of Special Session on “Machine Vision, Control and Navigation” at IEEE ISIE 2015. She has been incorporated to CONACYT National Research System in 2016.",institutionString:null,position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"Autonomous University of Baja California",institutionURL:null,country:{name:"Mexico"}}},coeditorTwo:{id:"58036",title:"Dr.",name:"Oleg",middleName:null,surname:"Sergiyenko",slug:"oleg-sergiyenko",fullName:"Oleg Sergiyenko",profilePictureURL:"https://mts.intechopen.com/storage/users/58036/images/5755_n.jpg",biography:"Oleg Yu. Sergiyenko received his BS and MS degrees from Kharkiv National University of Automobiles and Highways, Kharkiv, Ukraine, in 1991 and 1993, respectively. He received his PhD degree from Kharkiv National Polytechnic University in 1997.\nHe has written 81 papers in control systems, robot navigation, 3D coordinate measurement, and SHM. He is an editor of two books, holds two patents in Ukraine and Mexico, and is a reviewer for various journals. He participates as a reviewer and session chair in several IEEE conferences in different countries and holds several 'Best Presentation Awards.”\nFrom December 2004 to present, he is a full-time researcher and head of Applied Physics Department in Engineering Institute of Baja California Autonomous University, Mexico.",institutionString:null,position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"Autonomous University of Baja California",institutionURL:null,country:{name:"Mexico"}}},coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"1345",title:"Bioinformatics",slug:"technology-biomedical-engineering-bioinformatics"}],chapters:[{id:"54663",title:"Substructuring Method in Structural Health Monitoring",doi:"10.5772/67890",slug:"substructuring-method-in-structural-health-monitoring",totalDownloads:1282,totalCrossrefCites:1,totalDimensionsCites:1,hasAltmetrics:0,abstract:"In sensitivity-based finite element model updating, the eigensolutions and eigensensitivities are calculated repeatedly, which is a time-consuming process for large-scale structures. In this chapter, a forward substructuring method and an inverse substructuring method are proposed to fulfill the model updating of large-scale structures. In the forward substructuring method, the analytical FE model of the global structure is divided into several independent substructures. The eigensolutions of each independent substructure are used to recover the eigensolutions and eigensensitivities of the global structure. Consequently, only some specific substructures are reanalyzed in model updating and assembled with other untouched substructures to recover the eigensolutions and eigensensitivities of the global structure. In the inverse substructuring method, the experimental modal data of the global structure are disassembled into substructural flexibility. Afterwards, each substructure is treated as an independent structure to reproduce its flexibility through a model-updating process. Employing the substructuring method, the model updating of a substructure can be conducted by measuring the local area of the concerned substructure solely. Finally, application of the proposed methods to a laboratory tested frame structure reveals that the forward and inverse substructuring methods are effective in model updating and damage identification.",signatures:"Shun Weng, Hong-Ping Zhu, Yong Xia and Fei Gao",downloadPdfUrl:"/chapter/pdf-download/54663",previewPdfUrl:"/chapter/pdf-preview/54663",authors:[{id:"198741",title:"Associate Prof.",name:"Shun",surname:"Weng",slug:"shun-weng",fullName:"Shun Weng"},{id:"198763",title:"Prof.",name:"Hongping",surname:"Zhu",slug:"hongping-zhu",fullName:"Hongping Zhu"},{id:"198764",title:"Prof.",name:"Yong",surname:"Xia",slug:"yong-xia",fullName:"Yong Xia"},{id:"198765",title:"Prof.",name:"Fei",surname:"Gao",slug:"fei-gao",fullName:"Fei Gao"}],corrections:null},{id:"55607",title:"A New Method of SHM for Steel Wire Rope and its Apparatus",doi:"10.5772/intechopen.68148",slug:"a-new-method-of-shm-for-steel-wire-rope-and-its-apparatus",totalDownloads:1509,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Steel wire ropes often operate in a high‐speed swing status in practical engineering, and the reliable structural health monitoring (SHM) for them directly relates to human lives; however, they are usually beyond the capability of present portable magnet magnetic flux leakage (MFL) sensors based on yoke magnetic method due to its strong magnetic force and large weight. Unlike the yoke method, a new method of SHM for steel wire rope is proposed by theoretical analyses and also verified by finite element method (FEM) and experiments, which features much weaker magnetic interaction force and similar magnetization capability compared to the traditional yoke method. Meanwhile, the relevant detection apparatus or sensor is designed by simulation optimization. Furthermore, experimental comparisons between the new and yoke sensors for steel wire rope inspection are also conducted, which successfully confirm the characterization of smaller magnetic interaction force, less wear, and damage in contrast with traditional technologies. Finally, methods for SHM of steel wire rope and apparatus are discussed, which demonstrate the good practicability for SHM of steel wire rope under poor working conditions.",signatures:"Shiwei Liu, Yanhua Sun and Wenjia Ma",downloadPdfUrl:"/chapter/pdf-download/55607",previewPdfUrl:"/chapter/pdf-preview/55607",authors:[{id:"178404",title:"Associate Prof.",name:"Yanhua",surname:"Sun",slug:"yanhua-sun",fullName:"Yanhua Sun"},{id:"200684",title:"Dr.",name:"Shiwei",surname:"Liu",slug:"shiwei-liu",fullName:"Shiwei Liu"},{id:"200685",title:"MSc.",name:"Wenjia",surname:"Ma",slug:"wenjia-ma",fullName:"Wenjia Ma"}],corrections:null},{id:"55663",title:"Structural Damage Detection Based on Improved Multi-Particle Swarm Co-Evolution Optimization Algorithm",doi:"10.5772/intechopen.68385",slug:"structural-damage-detection-based-on-improved-multi-particle-swarm-co-evolution-optimization-algorit",totalDownloads:1025,totalCrossrefCites:0,totalDimensionsCites:2,hasAltmetrics:0,abstract:"This chapter presents an improved multi-particle swarm co-evolution optimization algorithm (IMPSCO) to detect structural damage. Firstly, IMPSCO is integrated with Newmark’s algorithm for damage detection and system identification, which just need few sensors. In addition, for reducing the searching parameters, a two-stage damage detection method based on modal strain energy and IMPSCO is presented. In order to validate the proposed method, a seven-story steel frame experiment in laboratory conditions is performed and a comparison is made between the proposed approach and genetic algorithm (GA). The results show that: (1) the proposed methods can not only effectively locate damage location but also accurately quantify the damage severity. Besides, it has excellent noise-tolerance and adaptability; (2) for integrating IMPSCO and Newmark’s algorithm, it implements only by using any kinds of structural time-series responses and the excitation force; (3) compared with genetic algorithm, IMPSCO is more efficient and robust for damage detection with a better noise-tolerance.",signatures:"Shaofei Jiang and Shenglan Ma",downloadPdfUrl:"/chapter/pdf-download/55663",previewPdfUrl:"/chapter/pdf-preview/55663",authors:[{id:"199255",title:"Prof.",name:"Shaofei",surname:"Jiang",slug:"shaofei-jiang",fullName:"Shaofei Jiang"},{id:"205301",title:"Dr.",name:"Shenglan",surname:"Ma",slug:"shenglan-ma",fullName:"Shenglan Ma"}],corrections:null},{id:"55249",title:"Operational Modal Analysis of Super Tall Buildings by a Bayesian Approach",doi:"10.5772/intechopen.68397",slug:"operational-modal-analysis-of-super-tall-buildings-by-a-bayesian-approach",totalDownloads:1496,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Structural health monitoring (SHM) has attracted increasing attention in the past few decades. It aims at monitoring the existing structures based on data acquired by different sensor networks. Modal identification is usually the first step in SHM, and it aims at identifying the modal parameters mainly including natural frequency, damping ratio and mode shape. Three different field tests can be used to collect data for modal identification, among which, ambient vibration test is the most convenient and economical one since it does not require to measure input information. This chapter will focus on the operational modal analysis (OMA), i.e. ambient modal identification of four super tall buildings by a Bayesian approach. A fast frequency domain Bayesian fast fourier transform (FFT) approach will be introduced for OMA. In addition to the most probable value (MPV) of modal parameters, the associated posterior uncertainty will be also investigated analytically. The field tests will be presented and the difficulties encountered will be discussed. Some basic dynamic characteristics will be investigated and discussed. The studies will provide baseline properties of these super tall buildings and provide a reference for future condition assessments.",signatures:"Feng-Liang Zhang and Yan-Chun Ni",downloadPdfUrl:"/chapter/pdf-download/55249",previewPdfUrl:"/chapter/pdf-preview/55249",authors:[{id:"202976",title:"Dr.",name:"Feng-Liang",surname:"Zhang",slug:"feng-liang-zhang",fullName:"Feng-Liang Zhang"},{id:"205623",title:"Dr.",name:"Yan-Chun",surname:"Ni",slug:"yan-chun-ni",fullName:"Yan-Chun Ni"}],corrections:null},{id:"54913",title:"Mooring Integrity Management: Novel Approaches Towards In Situ Monitoring",doi:"10.5772/intechopen.68386",slug:"mooring-integrity-management-novel-approaches-towards-in-situ-monitoring",totalDownloads:1504,totalCrossrefCites:3,totalDimensionsCites:8,hasAltmetrics:0,abstract:"The recent dramatic fluctuations in oil and gas prices are forcing operators to look at radically new ways of maintaining the integrity of their structures. Moreover, the life of old structures has to be extended. This includes the replacement of expensive periodic in-service inspections with cost-efficient structural health monitoring (SHM) with permanently installed sensors. Mooring chains for floating offshore installations, typically designed for a 25-year service life, are loaded in fatigue in a seawater environment. There is no industry consensus on failure mechanisms or even defect initiation that mooring chains may incur. Moorings are safety-critical areas, which by their nature are hazardous to inspect. Close visual inspection in the turret is usually too hazardous for divers, yet is not possible with remotely operated vehicles (ROVs), because of limited access. Conventional non-destructive techniques (NDTs) are used to carry out inspections of mooring chains in the turret of floating production storage and offloading (FPSO) units. Although successful at detecting and assessing the fatigue cracks, the hazardous nature of the operation calls for remote techniques that can be applied continuously to identify damage initiation and progress. Appropriate replacement plans must enhance current strategies by implementing real-time data retrofit.",signatures:"Ángela Angulo, Graham Edwards, Slim Soua and Tat-Hean Gan",downloadPdfUrl:"/chapter/pdf-download/54913",previewPdfUrl:"/chapter/pdf-preview/54913",authors:[{id:"78586",title:"Prof.",name:"Tat Hean",surname:"Gan",slug:"tat-hean-gan",fullName:"Tat Hean Gan"},{id:"178330",title:"Dr.",name:"Slim",surname:"Soua",slug:"slim-soua",fullName:"Slim Soua"},{id:"185485",title:"Ms.",name:"Ángela",surname:"Angulo",slug:"angela-angulo",fullName:"Ángela Angulo"},{id:"205582",title:"Mr.",name:"Graham",surname:"Edwards",slug:"graham-edwards",fullName:"Graham Edwards"}],corrections:null},{id:"54950",title:"Multi-Sensor Feature Extraction and Data Fusion Using ANFIS and 2D Wavelet Transform in Structural Health Monitoring",doi:"10.5772/intechopen.68147",slug:"multi-sensor-feature-extraction-and-data-fusion-using-anfis-and-2d-wavelet-transform-in-structural-h",totalDownloads:1335,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"In this chapter, a novel feature extraction and data fusion approach for structural damage detection and localisation is presented. This approach combines adaptive network-based fuzzy inference systems (ANFIS) and two-dimensional wavelet transform (2D-WT) technologies. Simultaneous multi-sensor feature extraction and data fusion based on 2D-WT is carried out by forming a 2D multivariate signal, which is used to analyse the structure vibration response by measuring all sensors jointly. Energy values obtained from two-level db3 wavelet decomposition are arranged in a so-called energy percentage matrix (EPM), which is taken as an input for the ANFIS. The system is further trained by defining its output as the structural condition represented by a condition index. A set of output index patterns are defined depending on the level of damage assessment performed. The proposed method was tested through experiments using a cantilever beam structure. The testing results showed that the method is successful in detecting and localising damage by vibration analysis in structural health monitoring.",signatures:"Ponciano Jorge Escamilla-Ambrosio, Xuefeng Liu, Juan Manuel\nRamírez-Cortés, Abraham Rodríguez-Mota and María del Pilar\nGómez-Gil",downloadPdfUrl:"/chapter/pdf-download/54950",previewPdfUrl:"/chapter/pdf-preview/54950",authors:[{id:"203558",title:"Dr.",name:"Abraham",surname:"Rodríguez-Mota",slug:"abraham-rodriguez-mota",fullName:"Abraham Rodríguez-Mota"},{id:"203701",title:"Dr.",name:"Ponciano",surname:"Escamilla-Ambrosio",slug:"ponciano-escamilla-ambrosio",fullName:"Ponciano Escamilla-Ambrosio"},{id:"203703",title:"Dr.",name:"X",surname:"Liu",slug:"x-liu",fullName:"X Liu"},{id:"203705",title:"Dr.",name:"J M",surname:"Ramirez-Cortes",slug:"j-m-ramirez-cortes",fullName:"J M Ramirez-Cortes"},{id:"205237",title:"Dr.",name:"Pilar",surname:"Gómez-Gil",slug:"pilar-gomez-gil",fullName:"Pilar 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This enormous availability of genomic information, in addition to its data availability, enables analysis of the complete genome of the species and imposes a growing demand for the understanding of several complex biological phenomena. Initially, the GWAS approach was applied in human genome studies involving entire sets of DNA from numerous individuals to find gene variations related to diseases such as asthma, diabetes, cancers, heart disease, and mental disorders. Nowadays the impact of GWS studies is not restricted only to the human genome to advances in the understanding of diseases. With the genome sequencing of several species and the availability of their complete genomes in large databases, other complex aspects can be investigated today by GWAS, such as evolutionary and phylogenetic relationships, characteristics of relevance for biodiversity conservation, and genetic improvement of plants and animals. Included in this book there are original and relevant works involving GWAS studies, carefully selected for the academic public. This book has been carefully designed to provide current and quality information to students and researchers of all levels who have an interest in Genome-Wide Association Studies and its applications.
",isbn:null,printIsbn:"979-953-307-X-X",pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,isNomenclature:!1,hash:"006916e730c66d3b84d3ec036f769e00",bookSignature:"Prof. Rafael Trindade Trindade Maia, Dr. Magnólia De Araújo Campos and Dr. Marco Antônio Alves Schetino",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11805.jpg",keywords:"Disorders, Deleterious Mutations, Association Mapping, Complex Diseases, Mega-Scale Genome Sequencing, Adaptative Fitness, Disease Resistance, Adaptative Alleles, Patterns of Inbreeding, Genome Phylogeny, Population Genetics, Population Structure",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"April 15th 2022",dateEndSecondStepPublish:"May 13th 2022",dateEndThirdStepPublish:"July 12th 2022",dateEndFourthStepPublish:"September 30th 2022",dateEndFifthStepPublish:"November 29th 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"a month",secondStepPassed:!0,areRegistrationsClosed:!1,currentStepOfPublishingProcess:3,editedByType:null,kuFlag:!1,biosketch:"Professor honoured as an academic reviewer with an interest in the areas of genetics, evolution, bioinformatics, and biotechnology. Academic editor of books and journals and author of 3 patents and 1 software registration. Dr. Rafael Trindade Maia received a master´s degree in Genetics, Conservation, and Evolutionary Biology from the National Institute of Amazonian Research, Brazil, in 2008, and a Ph.D. in Animal Biology from the Federal University of Pernambuco, Brazil, in 2013.",coeditorOneBiosketch:"Dr. Magnólia A. Campos is a researcher with experience in the Genetics and Plant genomics and the author of books and patents. She has a master's in Agronomy / Plant Breeding from the Federal University of Pelotas and a Ph.D. in Biological Sciences / Molecular Biology from the University of Brasília. Since 2008, she has been a Professor at the Federal University of Campina Grande (UFCG).",coeditorTwoBiosketch:"Dr. Marco Antônio Alves Schetino studied biological sciences at the Federal University of Viçosa (UFV), Brazil, he received a master´s degree in Genetics, Conservation, and Evolutionary Biology from the National Institute of Amazonian Research (INPA), Brazil, and a Ph.D. in Genetics from the Federal University of Minas Gerais (UFMG), Brazil. He is a researcher with experience in Genetics (with emphasis on Animal Genetics, Evolution, and major health areas) and philosophy of science areas.",coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"212393",title:"Prof.",name:"Rafael",middleName:"Trindade",surname:"Trindade Maia",slug:"rafael-trindade-maia",fullName:"Rafael Trindade Maia",profilePictureURL:"https://mts.intechopen.com/storage/users/212393/images/system/212393.jpg",biography:"Dr. Rafael Trindade Maia studied biological sciences at the Federal Rural University of Pernambuco, Brazil (2005). He received a master´s degree in Genetics, Conservation, and Evolutionary Biology from the National Institute of Amazonian Research, Brazil, in 2008, and a Ph.D. in Animal Biology from the Federal University of Pernambuco, Brazil, in 2013. He is currently an adjunct professor at the Center for the Sustainable Development for Semiarid (CDSA) at Federal University of Campina Grande (UFCG), Brazil. He has experience with population genetics, bioinformatics, molecular docking, and modeling and molecular dynamics of proteins. He works in science and biology education. Dr. Maia also leads the research groups Computational and Theoretical Biology (CTB) and Education in Sciences and Biology (ESB).",institutionString:"Federal University of Campina Grande",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"4",totalChapterViews:"0",totalEditedBooks:"3",institution:{name:"Federal University of Campina Grande",institutionURL:null,country:{name:"Brazil"}}}],coeditorOne:{id:"265397",title:"Dr.",name:"Magnólia De Araújo",middleName:null,surname:"Campos",slug:"magnolia-de-araujo-campos",fullName:"Magnólia De Araújo Campos",profilePictureURL:"https://mts.intechopen.com/storage/users/265397/images/system/265397.png",biography:"Magnólia A. Campos is a biologist, has a Masters in Agronomy / Plant Breeding from the Federal University of Pelotas and a PhD in Biological Sciences / Molecular Biology from the University of Brasília (2002). She had a total five years of experience in genomic sciences as a postdoctoral researcher at the Federal University of Lavras / Agronomic Institute (IAC). Since 2008, she has been a Professor at the Federal University of Campina Grande (UFCG). She has experience in the area of plant biotechnology, working mainly on the following topics: genomics, bioinformatics, tissue culture and plant cells, genetic transformation of plants, study of gene expression during plant-microbe interactions and expression of heterologous proteins in bacteria.",institutionString:"Federal University of Campina Grande",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"Federal University of Campina Grande",institutionURL:null,country:{name:"Brazil"}}},coeditorTwo:{id:"468502",title:"Dr.",name:"Marco Antônio",middleName:null,surname:"Alves Schetino",slug:"marco-antonio-alves-schetino",fullName:"Marco Antônio Alves Schetino",profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:"Dr. Marco Antônio Alves Schetino studied biological sciences at the Federal University of Viçosa (UFV), Brazil (2005), received a master´s degree in Genetics, Conservation, and Evolutionary Biology from the National Institute of Amazonian Research (INPA), Brazil, in 2008, and a Ph.D. in Genetics from the Federal University of Minas Gerais (UFMG), Brazil, in 2017. He is currently a Postdoc at the Federal University of Vales do Jequitinhonha e do Mucuri (UFVJM), Brazil. He has experience with Phylogenetic systematics, phylogeography, population genetics, evolution and conservation. Former professor at UNI-BH of the Biological Sciences and Ecology Course. Former substitute professor of the Bachelor of Science and Technology, BCT, at UFVJM.",institutionString:"Federal University of Vales do Jequitinhonha",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:null},coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"6",title:"Biochemistry, Genetics and Molecular Biology",slug:"biochemistry-genetics-and-molecular-biology"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"347259",firstName:"Karmen",lastName:"Daleta",middleName:null,title:"Ms.",imageUrl:"//cdnintech.com/web/frontend/www/assets/author.svg",email:"karmen@intechopen.com",biography:null}},relatedBooks:[{type:"book",id:"6694",title:"New Trends in Ion Exchange Studies",subtitle:null,isOpenForSubmission:!1,hash:"3de8c8b090fd8faa7c11ec5b387c486a",slug:"new-trends-in-ion-exchange-studies",bookSignature:"Selcan Karakuş",coverURL:"https://cdn.intechopen.com/books/images_new/6694.jpg",editedByType:"Edited by",editors:[{id:"206110",title:"Dr.",name:"Selcan",surname:"Karakuş",slug:"selcan-karakus",fullName:"Selcan Karakuş"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1591",title:"Infrared Spectroscopy",subtitle:"Materials Science, Engineering and Technology",isOpenForSubmission:!1,hash:"99b4b7b71a8caeb693ed762b40b017f4",slug:"infrared-spectroscopy-materials-science-engineering-and-technology",bookSignature:"Theophile Theophanides",coverURL:"https://cdn.intechopen.com/books/images_new/1591.jpg",editedByType:"Edited by",editors:[{id:"37194",title:"Dr.",name:"Theophile",surname:"Theophanides",slug:"theophile-theophanides",fullName:"Theophile Theophanides"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3161",title:"Frontiers in Guided Wave Optics and Optoelectronics",subtitle:null,isOpenForSubmission:!1,hash:"deb44e9c99f82bbce1083abea743146c",slug:"frontiers-in-guided-wave-optics-and-optoelectronics",bookSignature:"Bishnu Pal",coverURL:"https://cdn.intechopen.com/books/images_new/3161.jpg",editedByType:"Edited by",editors:[{id:"4782",title:"Prof.",name:"Bishnu",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"371",title:"Abiotic Stress in Plants",subtitle:"Mechanisms and Adaptations",isOpenForSubmission:!1,hash:"588466f487e307619849d72389178a74",slug:"abiotic-stress-in-plants-mechanisms-and-adaptations",bookSignature:"Arun Shanker and B. Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"2270",title:"Fourier Transform",subtitle:"Materials Analysis",isOpenForSubmission:!1,hash:"5e094b066da527193e878e160b4772af",slug:"fourier-transform-materials-analysis",bookSignature:"Salih Mohammed Salih",coverURL:"https://cdn.intechopen.com/books/images_new/2270.jpg",editedByType:"Edited by",editors:[{id:"111691",title:"Dr.Ing.",name:"Salih",surname:"Salih",slug:"salih-salih",fullName:"Salih Salih"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"117",title:"Artificial Neural Networks",subtitle:"Methodological Advances and Biomedical Applications",isOpenForSubmission:!1,hash:null,slug:"artificial-neural-networks-methodological-advances-and-biomedical-applications",bookSignature:"Kenji Suzuki",coverURL:"https://cdn.intechopen.com/books/images_new/117.jpg",editedByType:"Edited by",editors:[{id:"3095",title:"Prof.",name:"Kenji",surname:"Suzuki",slug:"kenji-suzuki",fullName:"Kenji Suzuki"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3828",title:"Application of Nanotechnology in Drug Delivery",subtitle:null,isOpenForSubmission:!1,hash:"51a27e7adbfafcfedb6e9683f209cba4",slug:"application-of-nanotechnology-in-drug-delivery",bookSignature:"Ali Demir Sezer",coverURL:"https://cdn.intechopen.com/books/images_new/3828.jpg",editedByType:"Edited by",editors:[{id:"62389",title:"PhD.",name:"Ali Demir",surname:"Sezer",slug:"ali-demir-sezer",fullName:"Ali Demir Sezer"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"872",title:"Organic Pollutants Ten Years After the Stockholm Convention",subtitle:"Environmental and Analytical Update",isOpenForSubmission:!1,hash:"f01dc7077e1d23f3d8f5454985cafa0a",slug:"organic-pollutants-ten-years-after-the-stockholm-convention-environmental-and-analytical-update",bookSignature:"Tomasz Puzyn and Aleksandra Mostrag-Szlichtyng",coverURL:"https://cdn.intechopen.com/books/images_new/872.jpg",editedByType:"Edited by",editors:[{id:"84887",title:"Dr.",name:"Tomasz",surname:"Puzyn",slug:"tomasz-puzyn",fullName:"Tomasz Puzyn"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"30757",title:"HPV Bioinformatics: In Silico Detection, Drug Design and Prevention Agent Development",doi:"10.5772/27456",slug:"hpv-bioinformatics-in-silico-detection-and-prevention-agent-development",body:'\n\t\tViral infection is a very serious threat to humanity. It causes malicious diseases, such as HIV/AIDS, dengue, and Avian Influenza, therefore, novel method in virology to combat the viral infection is necessary. Bioinformatics provides outstanding tools for developing vaccines, PCR primers, mutation detection and drugs based design on genetic engineering principles. Those tools are mostly freeware. Algorithm from the computer science has made major contribution to them. Bioinformatics experiment greatly reduces the cost and time in wet laboratory experiment. Our lab has successfully designed PCR primers, vaccine, and mutations prediction. The vaccine design is elaborated for Dengue and HPV. The design has BLAST homology of more than 90%, and RSMD value of 0.1. Those data shown, that the design have identical structure with the native viral protein.
\n\t\t\tHowever, their efficacy should be verified in the wet laboratory experiment. The future of medicine will greatly be shaped by advancement in bioinformatics (Tambunan et al, 2010). Moreover, we need to emphasize the needs to understand more about HPV genome and its protein expression.
\n\t\t\tThe human papillomavirus (HPV) is a family of sexually transmitted, double-stranded DNA viruses with over 100 different genotypes identified till date. It is associated with many different types of cancers including cervical, vaginal, head and neck, penile and anal cancer. With approximately 450,000 newly diagnosed cases each year and a 50% mortality rate, cervical cancer is the second most common cause of cancer-related death in women worldwide and it is almost always associated with HPV. Cervical cancer is the most common cancer of women in most developing countries, where it may account for as many as one fourth of female cancers.
\n\t\t\tHPV genotypes are divided into the low risk and high risk categories based on the spectrum of lesions they induce. The low-risk types induce only benign genital warts and include HPV 6 and 11. The high-risk group containing HPV 16, 18, 31, 33, 45 and 56 is associated with the development of anogenital cancers and can be detected in 99% of cervical cancers, with HPV16 found in 50% of cases.
\n\t\t\tA consistently effective and safe treatment for HPV infections is currently not yet available. Present therapeutic options are more directed at surgical eradication and/or by destroying malignant lesions via physical or chemotherapeutical intervention. A majority of these treatments have been developed empirically, few have been thoroughly tested, but none of them are completely satisfactory. In attemps to find additional drugs in the treatment of cervical cancer, inhibitors of the histone deacetylases have received much attention due to their low cytotoxic profiles (Tambunan et al, 2010).
\n\t\t\tElaborating a methodological comparison between our computational approaches with the other bioinformatics labs. We will not compare our method directly with wet labs, because our method is developed to supplement it. Our objective is to find the optimal in silico HPV therapeutic design, and implement it in the wet laboratory.
\n\t\tThis section will elaborate more on HPV Genome Annotation. The conserved region of HPV Genome needs to be annotated, in order to design a useful detection tools. Conventional method of detecting cervical cancer is done by carrying out cytological examination, which is more widely known as Pap smear. Due to rapid advancement of molecular biology, molecular based diagnostic and early detection methods on cervical cancer has been highly developed to replace conventional method of detection. Examples include polymerases chain reaction (PCR) and hybrid capture 1, 2 and 3. Hybrid capture 1 is liquid hybridization assaying method designed to detect 14 types of HPV, 9 of which are high risk (type 16, 18,31, 33, 35, 45, 51, 52, 56), while the other 5 are low risk (type 6, 11, 42, 43, and 44). Hybrid capture 2 is a development of hybrid capture 1 which uses microtitre plates instead of tubes and is capable of detecting four additional types of viral oncogenic (type 39, 58, 59, and 68) (Clavel et al, 1998). Hybrid capture 3, similar to previous hybrid capture tests, relies on the formation of target HPV DNA-RNA probe heteroduplexes during the hybridization step in specimens containing sufficient HPV DNA. The chemiluminescent detection of these hybrids is by adding an alkaline phosphatase-conjugated monoclonal antibody, specific to the DNA-RNA complexes with dioxetane substrate in a 96-well enzyme-linked immunosorbent assay format (Lorinze and Anthony, 2001). Most of the above mentioned methods are spesific, sensitive, reliable, and easy to perform. Moreover, its routine application has been very much improved by the use of non-radioactive enzyme immunoassay detection procedure (Clavel et al, 1998). Modification of the Hybrid Capture method is expected to be achieved through the use of a customized oligonucleotide probe able to detect multiple high-risk HPV infection.
\n\t\t\t\tThere are 10 genes encoded in HPV genome. These gene may be classified into two groups, namely Gene E (early) which encodes regulatory proteins and Gene L (late) which encodes structural proteins. The region of L1 and L2 of Gene L is responsible for encoding capsid protein to be used as DNA envelope in HPV. The capsid protein will serves as protection system to HPV genetic materials. It is generally assumed that there are many nucleotides sequence in the region of L1 and L2 that is conserved throughout the evolution process of HPV (Dahlgren, 2005).
\n\t\t\t\tMendez-Tenorio and his group were using DNA fingerprinting. Identification of microorganisms by whole genome DNA fingerprinting was tested “in silico”. 94 HPV genome sequences were submitted to virtual hybridization analysis on a DNA chip with 342 probes. This Universal Fingerprinting Chip (UFC) constitutes a representative set of probes of all the possible 8-mer sequences having at least two internal and non-contiguous sequence differences between all them. A virtual hybridization analysis was performed in order to find the fingerprinting pattern that represents the signals produced for the hybridization of the probes allowing at most a single mismatch. All the fingerprints for each virus were compared against each other in order to obtain all the pairwise distances measures. A match-extension strategy was applied to identify only the shared signals corresponding to the hybridization of the probes with homologous sequences between two HPV genomes. A phylogenetic tree was constructed from the fingerprint distances using the Neighbor-Joining algorithm implemented in the program Phylip 3.61. This tree was compared with that produced from the alignment of whole genome HPV sequences calculated with the program Clustal_X 1.83. The similarities between both trees are suggesting that the UFC-8 is able to discriminate accurately between viral genomes. A fingerprint comparative analysis suggests that the UFC-8 can differentiate between HPV types and subtypes ( Méndez-Tenorio
Kaladhar and his group are working on annotation HPV-92 genome. Most of the biologists focus to explore innovations of their research in faster rate using developments in Information technology. The gene identification, characterization and modeling of the proteins in HPV 92 is done using bioinformatics tools. A complete genome of HPV-92 with NCBI’s accession number NC_004500 was submitted to FGENES V0, a viral gene prediction server, predicts six genes. These six genes are characterized as E6 oncoprotein, E7 oncoprotein, E1 Replication protein, E2 Regulatory protein, L1 major capsid protein and L2 minor capsid protein. Isoelectric points and Molecular weights of all the six proteins vary largely and the modeled structures are shown. The research can provide characterization and modeling of genome which can further implemented in drug designing methods using bioinformatics tools (Kaladhar et al, 2010).
\n\t\t\t\t\tEom and his group has interesting approach towards mapping HPV genome. They are using genetic mining algorithm. Classifying the type of HPV is very important to the treat of cervical cancer. The machine learning approach to mine the structure of HPV DNA sequence for effective classification of the HPV risk types has been introduced. The most informative subsequence segment sets and its weights with genetic algorithm to classify the risk types of each HPV has been determined and learnt. To resolve the problem of computational complexity of genetic algorithm, distributed intelligent data engineering platform based on active grid concept called IDEA@Home was used. The proposed genetic mining method, with the described platform, shows about 85.6% classification accuracy with relatively fast mining speed (Eom et al, 2004).
\n\t\t\t\t\tLee and his groups are using in silico DNA microarry for detecting HPV genome. DNA microarrays are widely used techniques in molecular biology and DNA computing area. It consists of the DNA sequences called probes, which are DNA complementaries to the genes of interest, on solid surfaces. And its reliability seriously depends on the quality of the probe sequences. Therefore, one must carefully choose the probe sets in target sequences. The probe design for DNA microarrays was formulated as the multi-objective optimization problem. Multi-objective evolutionary approach was proposed, which is known to be suitable for this kind of optimization problem. Since a multi-objective evolutionary algorithm can find multiple solutions at a time, thermodynamic criteria was used to choose the most suitable one. For the experiments, the probe set generated by the proposed method is compared to the sequences used in commercial microarrays, which detects a set of Human Papillomavirus (HPV). The comparison result supports that the approach can be useful to optimize probe sequences (Lee et al, 2004).
\n\t\t\t\tThe aim of our study was to determine the conserved regions of late genes L1 and L2 from 74 sequenced and published HPV genome (Icenogle, 1995). The result was used to predict candidate template for oligonucleotide probes that are specific on types of HPV, which cause cervical cancer. Nevertheless, the spesific purpose of this study is to design primer that is able to detach on the open reading frame region and also to develop a new assay for the detection of high risk HPV DNA.
\n\t\t\t\t\tThis study was carried out to determine the conserved regions of late genes from sequenced HPV types. HPV genome sequences were collected from the Los Alamos National Laboratory
Finally, 7 selected conserved regions were examined for secondary structures using NetPrimer program. From this operation, only region 52 (5’-ACAGGCTATGGTGCTATGGA-3’) met the criteria to be used as an oligonucleotide primer (\n\t\t\t\t\t\t\tTambunan et al, 2007\n\t\t\t\t\t\t).
\n\t\t\t\t\tOligonucleotide primers are considered based upon certain properties, namely: they must not have potential secondary structures such as hairpins or dimmers have a GC content of 45-60%; have a Tm between 52-58%; their 5\' ends stability has to be greater than the stability of their 3\' ends; be 17-25 nucleotides in length. From NetPrimer analysis results of seven regions, only region 52 meet with the above mentioned criteria. with a NetPrimer rating of 100 (maxium) (\n\t\t\t\t\t\t\tTambunan et al, 2007\n\t\t\t\t\t\t). The result is shown in table 1. Based on sequence similarity, 62 conserved regions were found. Out of the 62, 7 regions were then used as templates for primers used in detection of high and low risk HPV. From the 7 template candidates, only one met the criteria to be used as an oligonucleotide primer, namely region 52. From the study, region 52 is predicted to be selective to be used in the detection of oncogenic Human
A new paradigm of vaccine design is now emerging, following essential discoveries in immunology and the development of bioinformatics tools for T-cell epitope prediction from
\n\t\t\t\t\n\t\t\t\t\t\t\t\t | \n\t\t\t\t\t\t
Conserved regions selected as templates
primary protein sequences. One rationale for this new paradigm is that following exposure to a pathogen, epitope-specific memory T-cell clones are established. These clones respond rapidly and efficiently upon any subsequent infection, elaborating cytokines, killing infected host cells, and marshalling humoral and cellular defences against the pathogen. The most efficient immune response to some pathogens is derived from a number of different T cells that respond to an ensemble of pathogen-derived short peptides called epitopes. Whether an immune response is directed against a single immunodominant epitope or against many epitopes, the generation of a protective immune response does not require the development of T-cell memory to every possible peptide in the entire pathogen. T-cell response to the ensemble of epitopes, not the whole pathogen, is the source from which a protective immune response is derived. Similarly, if an individual is previously exposed to a language, upon hearing just a few words of that language he/she will usually recognize, for example, that French or English is being spoken. Complete mastery of the language is not required for this recognition. Using this analogy to describe epitopes, one could say that they are pathogen-specific \'words\' that alert the immune system to the presence of a pathogen. It is now possible to envisage the design of vaccines based on an ensemble of epitopes (a string of words, a few sentences, a paragraph, or a chapter) derived from the genome of a pathogen, using tools that have been developed in the field of immuno-informatics (De Groot et al, 2002 ). Knowledge about immunology is crucial in designing vaccine. Immunoinformatics, which is a branch of Bioinformatics, is a flourishing field.
\n\t\t\t\tThe significant breakthrough in HPV vaccine research was accomplished, when capsid protein L1 and L2 was found to be able to assemble themselves to be Virus Like Particles (VLP) during cell expression. VLP is very similar to the native HPV particle, and it includes the conformation epitope that induce the viral neutralization antibody. This is very crucial for the immune system, in order to detect VLP as viral infection, and giving the proper response. Because VLP is coreless and didn\'t contain the viral DNA, then it is expected that it won\'t create infection. The produced VLPs are type 6,11, 16, 18, 31, 33, 35, 39, 45,and 58). One VLP Chimeric (cVLP) model has sucessfully induced Mice Cytotoxic T Cell, by joining E6 and E7 capsid protein in the VLP. Some scientist believes, that cVLP has huge potentials to be utilized as infection prevention (Kolls et al, 2000).
\n\t\t\t\tProtein must be folded like its native conformation, in order to be activated as mature protein. The protein modification into its native conformation is called as post-translation modification. The polypeptide chain, which consisted of more than 200 residues, is usually folded into two or more globular domain. Most of the domain has 100 until 200 amino acid residues, and having diameter of ~25 Å (Voet et al, 1995).
\n\t\t\t\tDNA vaccine is designed using choice of a suitable expression vector, ensuring optimal expression by codon optimization, engineering CpG motifs for enhancing immune responses and providing additional sequence signals for efficient translation. DNA vaccines have been one of the latest developments in vaccine technology. DNA vaccines are essentially plasmids capable of expressing an antigenicpeptide in the host. These expressed proteins are recognized as foreign in the cells of the body. They are processed by the host cells and displayed on their surface to alert the immune system and trigger body’s immune responses. DNA vaccines have become anattractive alternative to conventional methods due to the fact that it can elicit sustained cell-mediated as well as humoral immun responses, which is very much important in combating pathogenic organisms, especially intracellular pathogens. Vaccine efficacy can be assessed by correlating the vaccine’s immunogenicity such as its ability to induce CD8+ or CD4+ T cells to the HPV oncoproteins with its ability to protect vaccinated animals against formation of tumors or to cause clearance of already established tumors. Recently several techniques like optimizing codons, CpG optimization and promoter and resistance gene insertion have been tried to enhance the immunogenicity of DNA vaccines (Gupta et al, 2009).
\n\t\t\t\tGupta and his group are working with DNA vaccine. There is a need to develop a new prophylactic DNA vaccine, which can work against different strains of HPVs and may lead to protection of cervical cancer against new pandemicviruses. Potential prophylactic DNA vaccine has been designed by using all the consensus epitopic sequences of HPVs L2 capsid protein and performed in silico cloning of multiepitopic antigenic DNA sequence in pVAX-1 vector. Immunogenicity of vaccine has been enhanced by techniques like codon optimization, engineering CpG motifs, introducing promoters and co-injection with plasmids expressing immune-stimulatory molecules (Gupta et al, 2009).
\n\t\t\t\tUnlike others, our laboratory is using chimeric protein for designing the vaccine. The cVLP HPV-16 ANN1, ANN2, HMM1, and HMM2 in silico vaccine design were discovered. The BLAST test towards them was generating 96% identity with native L1 HPV-16 protein. Therefore, it is expected that the vaccines could cause same level of immunogenicity with the native protein. The Ramacandran Plot of them showed that the disallowed region plot of non-glycine residue was less than 15%. Henceforth, the quality of the vaccine could be structuraly good. The VAST test toward them showed the RMSD of 0,1 Å, which shows that they have a high structural similarity.
\n\t\t\t\t\tBased upon in silico predition, it was found that post-translational modification could occur at cVLP. During the formation of cVLP, the possible occurred post-translational modification is N-Glicocylation. It is because this modification has N-Xaa-S/T motif which found at our in silico detection method. Although it is expected to happen, it is predicted that this modification wouldn\'t affect the tability of the cVLP, because its epitope did not affected.
\n\t\t\t\t\tChimeric virus like particles (cVLP) has been developed as vaccine candidate for preventing cervical cancer. cVLPs are improvement of Virus Like Particles (VLP) by substituting the epitope of L1 HPV -18 and -52 protein to L1 HPV -16 protein. They are ANN1, ANN2, HMM1, and HMM2. The impact of post translation modification will be determined. Based on In Silico study, the dominant post translation modification is glycosylation (\n\t\t\t\t\t\t\tTambunan et al 2007\n\t\t\t\t\t\t).
\n\t\t\t\t\tHowever, the next step is to develop in silico plasmid vector for expressing cVLP at the euchariotic host cell. The necessary step is to conduct in vitro experimentation to construct the cVLP HPV L1 at the proper host cell. After the cVLP has sucessfully produced, we could conduct in vivo research to determine the immunogenicity of cVLP at the animal testing, for example at rabbits or mice. Figure 1 shows the amino acid sequence of our vaccine design, while figure 2 shows its 3D visualization.
\n\t\t\t\t\tThe sequence of Our ANN1 cVLP L1 HPV Vaccine design.
Our ANN1 cVLP L1 HPV Vaccine Design. The vaccine was visualized by using MacPymol application. The ribbons inside the chains are the vaccine backbone (
This section will elaborate more on Drugs design. Knowledge about protein receptor-inhibitor interaction is very important for designing drugs. Computational Chemistry is the major supporting science in it. The structural modification of histones is playing important roles in the knowledge of HPV drug design.
\n\t\t\t\tHowever, we are going to explain more about drug design biochemistry background. The structural modification of histones is regulated mainly by acetylation/deacetylation of the N-terminal tail and is crucial in modulating gene expression, because it affects the interaction of DNA with transcription-regulatory non-nucleosomal protein complexes. The balance between the acetylated/deacetylated states of histones is mediated by two different sets of enzymes: histone acetyltransferases (HATs) and histone deacetylases (HDACs). HATs prefentially acetylate specific lysine substrates among other non-histones protein sub- strates and transcription factors, affecting DNA-binding properties and, in turn, altering gene transcription. HDACs restore the positive charge on lysine residues by removing acetyl groups and thus are involved primarily in the repression of gene transcription by compacting chromatin structure. Therefore, open lysine residues attach firmly to the phosphate backbone of the DNA, preventing transcription. In this tight conformation, transcription factors, regulatory complexes, and RNA polymerases cannot bind to DNA Acetylation relaxes the DNA conformation, making it accessible to transcription machinery. High levels of acetylation of core histones are seen in chromatin-containing genes, which are highly transcribed genes; genes that are silent are associated with low levels of acetylation. Inappropriate silencing of critical genes can result in one or both hits of tumor suppressor gene inactivation in cancer. Members of the classical HDAC family fall into two different phylogenetic classes, namely class I and class II. The class I HDACs (HDAC1, HDAC2, HDAC3, and HDAC8) are most closely related to the yeast (
The inhibition of HDAC activity by a specific inhibitor induces growth arrest, differentiation, and apoptosis of transformed cells as well as several cancer cells (Subha et al, 2008). Recent studies were directed to investigate the molecular effects of HDAC inhibition on cervical carcinoma cells as well as on primary human foreskin keratino- cytes, separately immortalized with amphotropic retroviruses that carry the open reading frames of HPV 16 E6, E7 or E6/E7. In these experiments one could show that E6/E7 oncogene function of human papillomavirus can be completely bypassed by HDAC inhibition. Both malignant and immortalized HPV 16/18-positive cells became blocked in G1/S transition despite ongoing viral gene expression. G1 arrest was accompanied by a down-regulation of cyclin D and cyclin A and a concomitant up regulation of the cyclin kinase inhibitors (CKI) p21 and p27. Binding of both CKIs led to a complete block of the cyclin-dependent kinase (cdk2) activity and in turn prevented binding of E7. This was intriguing with respect to the reversibility of HPV transformation process, since it is thought that the abrogation of the growth inhibitory function of p21 and p27 through E7 represents a key event in HPV-induced carcinogenesis. HDAC inhibitors also trigger pRb degradation, while E2F expression remained unaffected. pRb degradation is an E7- specific phenomenon, since in E6-positive cells pRb only became hypophosphorylated. The presence of E2F under cell cycle arrest led to a classical “conflict situation” which finally induced apoptosis (Finzer et al, 2001; Finzer et al, 2002; Finzer et al, 2004). Hence, the knowledge how the transforming potential of HPV can be bypassed without switching off viral transcription could open new therapeutical perspectives for the treatment of cervical cancer (Acharya et al, 2006). The drug design on HPV are mainly tampering with the reactivity of E(early) protein.
\n\t\t\t\tRehmi and his group are mainly working on E2 proteins. The E2 protein from HPV 16 was selected as a molecular target and its known structures were exploited for broad scope of “hits” to be identified in the screening process. They compared both structure-based and ligand-based design approaches for virtual screening. Databases enriched in natural compounds were used for virtual screening based on molecular docking. In this study, they identified novel classes of HPV inhibitors by means of a structure-based drug-design protocol involving Pharmacophore based virtual screening with molecular docking simulation (Rehmi et al, 2009).
\n\t\t\t\t\tBaleja and his group have different approach, because they are using E6 protein as vaccine template. The E6 protein from the high-risk HPV types represents an attractive target for intervention because of its roles in viral propagation and cellular transformation. E6 functions in part by interaction with human cellular proteins, several of which possess a helical E6-binding motif. The role for each amino acid in this motif for binding E6 has been tested through structure determination and site-directed mutagenesis. These structural and molecular biological approaches defined the spatial geometry of functional groups necessary for binding to E6. This E6-binding information (the E6-binding pharmacophore) was transferred into a three-dimensional query format suitable for computational screening of large chemical databases. Compounds were identified and tested using in vitro and cell culture-based assays. Several compounds selectively inhibited E6 interaction with the E6-binding protein E6AP and interfered with the ability of E6 to promote p53 degradation. Such compounds provide leads for the development of new pharmacologic agents to treat papillomavirus infections and their associated cancers (Baleja et al, 2006).
\n\t\t\t\tOur approach was done by Identification of a better
In this paper, we present homology models of six
The certain types of HPV are involved in the development of cervical cancer. In attempts to find additional drugs in the treatment of cervical cancer, inhibitors of the histone deacetylases (HDAC) have received much attention due to their low cytotoxic profiles and the E6/E7 oncogene function of human papilomavirus can be completely by passed by HDAC inhibition. The histone deacetylase inhibitors can induce growth arrest, differentiation and apoptosis of cancer cells. HDAC class I and class II are considered the main targets for cancer. Therefore, the six HDACs class II was modelled and about two inhibitors (SAHA and TSA) were docked using AutoDock 4.2, to each of the inhibitor in order to identify the pharmacological properties. Based on the results of docking, SAHA and TSA were able to bind with zinc ion in HDACs models as a drug target. SAHA was satisfied almost all the properties i.e., binding affinity, the Drug-Likeness value and Drug Score with 70% oral bioavailability and the carbonyl group of these compound fits well into the active site of the target where the zinc is present. Hence, SAHA could be developed as potential inhibitors of class II HDACs and valuable cervical cancer drug candidate (Tambunan et al, 2010).
\n\t\t\t\t\tStructure of SAHA and TSA. SAHA and TSA are hydroxamic acid derivatives that can be HDAV inhibitors.
Each ligand shows different affinity with class II HDAC, for example SAHA compound show best affinity with HDAC 5 based on Autodock calculation, and HDAC 10 based on APBS calculation. Whereas the same compound was found to be rank 2 with HDAC7 (-7.42 kcal/mol) based on AutoDock calculation and HDAC6 (-213.60 kJ/mol) based on APBS calculation, and rank 3 (-6.72 kcal/mol) with HDAC10 (AutoDock) and HDAC7 (-203.21 kJ/mol, APBS). Local free binding energy obtained from AutoDock of
The further descriptor analysis and the toxicity prediction helped in the identification of the better inhibitor. Drug Score and the Drug-Likeness are the two properties that are important for considering a compound to become a successful drug. TSA had a drug score of 0.37 and drug likeness property score of 1.24, which is higher than those for SAHA with respective scores of 0.35 and -8.87. The molecular weight of SAHA was 264.32 g/mol and that of TSA was 302.37 g/mol (Table 1), between the preferred range of molecular weight for drug
\n\t\t\t\t\tStructures of docked SAHA with
Structures of docked TSA with HDAC Class II
likeness property (160-480 g/mol). TSA was having tumorigenic property. SAHA was the compound that had the acceptable range for toxicity risk. These values were also taken into account to decide the best inhibitor. Thus, SAHA was the best drug candidate than TSA and also found to possess better global binding affinity score. The ADME-TOX box results showed that the SAHA has an oral bioavailability of more than 70% i.e., good solubility and stability. It acts as a non-substrate and non-inhibitor of P-gp. SAHA does not undergo significant first-pass metabolism.
\n\t\t\t\t\tThe three-dimensional models for six class II histone deacetylases (HDAC4, HDAC5, HDAC6, HDAC7, HDAC9, and HDAC10), which were built using homology modeling and validated by bioinformatics techniques and by comparison to an X-ray structures, were docked to SAHA and TSA. Our studies provide the structural view of the catalytic domain of a class II HDAC and reveal for this subclass specific features: (i) novel zinc binding motif that is likely to be involved in substrate binding and/or protein-protein interactions and may provide a site for modulation of activity, and (ii) a unique active site topology in catalytic activity. SAHA and TSA predicted to inhibit the class II HDACs are effective to all forms HDACs. SAHA was satisfied almost all the properties i.e., binding affinity scores of SAHA in the six class II HDAC enzymes was -156.94 kJ/mol, -171.77 kJ/mol, -213.60 kJ/mol, -203.21 kJ/mol, -179.29 kJ/mol & -263.15 kJ/mol respectively, the Drug Likeness value (1.24) and drug score (0.37) with 70% oral bioavailability and the carbonyl group of these com- pounds fits well into the active site of the target where the zinc is present. Hence, SAHA could be developed as potential inhibitors of class II HDACs and valuable anti cancer agents.
\n\t\t\t\tThe IT industry has provided strong and robust computing power, with low cost expediture. Nowadays, a powerful low cost multiprocessor computers are available,which made the modeling of complicated proteins and sophisticated drug design possible. The major computer operating system, such as MacOSX, Linux, and Windows are already supporting open source bioinformatics software. They could do the functionalities of the commercial software, with the same robustness. Nowadays, the field of bioinformatics is growing. The In Silico (Bioinformatics) experiment will be considered as important as wet experiment by biologist and/or biotechnologist. The In silico approach did not designed to replace wet experiment, but it’s in order to supplement it. Open source implementation will help bioinformaticians to solve viral threat in efficient and effective manner. There will be more robust bioinformatics tools available in the future for solving crucial virology related problems. Our laboratory has successfully designing primer and vaccine for therapeutics. However, the efficacy of the design must be proven in the wet labs experiment. Synthesizing them by using latest molecular biology instrument is crucial for progressing towards clinical trial. Conducting it will require us to form strong cooperation with faculty of medicine in our university. We already have cooperation with them, and will verify our design in the future.
\n\t\t\tHPV Bioinformatics is a growing and developing field. Our labs has sucessfully developed HPV Genome detection, Vaccine, and drugs design. We found 7 conserve region candidates for PCR design, HPV L1 cVLP for vaccine design, and SAHA/TSA drug design. In silico PCR Primer, Vaccine, and drugs design are possible with the newest development in algorithm and programming. Our labs and others has successfully developed them. The next challenges would be implementing them in the wet laboratory research.
\n\t\tThis publication is supported by Universitas Indonesia. The authors are grateful to to Dr. Ridla Bakri, Chairman of Department of Chemistry, Faculty of Mathematics and Natural Science, University of Indonesia for his support.
\n\t\tExtracellular vesicles (EVs) are traditionally classified into three types: exosomes (Exo), microvesicles (MVs), and apoptotic vesicles. Several theories exist on how tumor cells alter their neighboring cells and matrix ultimately changing their behavior into an invasive one. This typically would involve the transport of materials from tumor cells to their adjacent surroundings. These materials include a wide range of soluble cytokines, RNA species, enzymes, and proteins. Most of which are carried in nano-sized carriers such as EVs. EVs are classified according to their size and the mechanism of genesis. The first class of EVs known as MVs or when secreted from cancer cells, are called oncosomes [1]. MVs formation is originated by the outward budding of the cell surface at specific regions along the plasma membrane enriched with high concentrations of lipids, such as cholesterol and glycosphingolipids, and proteins such as Flotillin-1 and 2 [2]. Exo represent the second major class of EVs [3]. They are formed when multivesicular bodies (MVBs) in the endo-lysosomal pathway accumulate intraluminal vesicles (ILVs) that consist of proteins and nucleic acids. Exo are smaller in size and range from 30 to 50 nm.
EVs can function in an autocrine, paracrine, and even endocrine fashion, and were shown to impact various cancer cell phenotypes, increasing their cell growth and promoting metastasis [4]. This secretome is released into the microenvironment and acts as cell-cell communicators. Tumor derived Exo (TDE) has appeared as imperative facilitators in cancer initiation, progression, metastasis, host immune suppression, and drug resistance [5]. TDE typically consists of high sphingolipids and cholesterol contents that contain major histocompatibility complex (MHC) molecules, heat shock proteins, and tetraspanin (CD63, CD81, and CD9). Additionally, tumor antigens such as Mart1, gp100, TRP, and Her2-neu have been discovered in TDE [5]. TDE also contains surface and soluble proteins and RNA species such as mRNAs and miRNAs. mRNAs conveyed in EVs result in proteins synthesis in target cells, while miRNAs alter their gene expression [6]. The
Tumor development is a multistep process that starts by cellular reprogramming of cells to acquire the hallmarks of cancer cells to gain and maintain abnormal growth and invasive capacity [8]. The complex process of tumor formation and spreading additionally requires a rewiring of the surrounding stromal cells. This can be induced by intrinsic cell events such as genetic or epigenetic aberrations or by external factors from direct or indirect cell communication [9]. In cancer, EVs especially Exo, have been shown to be essential for various steps during tumor initiation and progression. EVs disrupt signaling and gene expression regulation in the recipient cell by horizontally transferring bioactive chemicals between cancer cells and the surrounding stroma. As a result, malignant cells can change the phenotype of surrounding benign cells to one that supports tumor growth and metastasis, creating a favorable environment for cancer progression and spread. EVs play several roles in priming the surrounding environment preparing it for metastasis and invasion. The role of EVs in promoting tumor progression has been elucidated in studies on mixed populations of EVs. The function of EVs largely depends on their bioactive cargo, in particular the shuttling of tumor-specific proteins to the surrounding cells. While researchers have mainly studied the RNA content of EVs, however, the focus is starting to shift towards the EVs proteome [10].
The protein content of MVs within mixed populations of EVs was discovered to be significantly diverse from that of the Exo proteome, and is supplemented in proteins involved in microtubule, actin, and cytoskeleton networks, ARF6, its effector phospholipase D2, and parts of the endosomal sorting complex required for transport family (ESCRT-I) [11]. By transporting these molecules, MVs can impact nearby tumor cells and stromal cells.
One example in which MVs shed by the cancer cells were shown to enhance tumor cell proliferation is in multiple myeloma. This effect was shown to be related to the amelioration of the Extracellular Matrix Metalloproteinase Inducer (EMMPRIN/CD147) on the tumor MVs. This protein is known to be overexpressed in solid tumors, some lymphomas, and leukemias [12]. Another study in breast cancer cells found that the highly glycosylated version of EMMPRIN exists in high quantities in breast cancer cell-derived MVs and enhances tumor invasion through activation of p38/MAPK signaling [11]. Interestingly, it was found that MVs from patient Blood with metastatic breast cancer had a similar high-EMMPRIN expression, along with the tumor marker Mucin-1 (MUC1/CA 15-3) [11]. Additionally, the truncated oncogenic form of the epidermal growth factor receptor (EGFR), EGFRvIII, commonly expressed in aggressive brain tumor cells, is associated with pro-tumorigenic tumor–tumor crosstalk via MVs. It was discovered that EGFRvIII was present in MVs released by U373 glioma cells, allowing them to transfer malignant features from highly aggressive tumor cells to the more benign tumor cells, EGFRvIII-negative, thereby facilitating their oncogenic transformation [11]. Hence, MVs are convenient communicators within the TME, as they can either mediate the horizontal transfer of oncogenic material or activate oncogenic signaling pathways in neighboring cancer cells, enhancing their survival, proliferative, and angiogenic potential and triggering their transformation into an aggressive phenotype.
Alongside the tumor–tumor communication, MVs were proven to facilitate the crosstalk between the tumor and its surrounding stroma and immune cells which ultimately leads to cancer immune evasion. In breast cancer cells, the secretion of both tumor MV and TDE induced the expression of Wnt5a in tumor-associated macrophages. Macrophage Wnt5a promoted ß-catenin-independent Wnt signaling in breast cancer cells when delivered by macrophage-derived MVs and Exo, resulting in enhanced tumor invasion. This shows how EV-based cell-cell communication can drive tumor-associated immune cells to stimulate tumor growth [11]. MVs-enriched preparations induced the differentiation of monocytes producing anti-inflammatory cytokines such as IL-10. In line with this, early stimulation with tumor MV triggered macrophage polarization towards an anti-inflammatory phenotype with decreased anti-tumor cytotoxic potential. Additionally, as T cells represent the first line of the immune defense, tumor cells appear to suppress T cell activity and diminish antitumoral immune response via MVs-mediated cell-cell communication. For instance, leukemia-derived MVs deliver miRNAs to T cells, which alters T cell phenotype [13] (Figure 1). Moreover, MVs released by irradiated breast cancer cells were shown to carry abundant immune-suppressive proteins, such as programmed cell death ligand 1 (PD-L1) which inhibited cytotoxic T cell activity and enabled tumor growth (Figure 1)[15].
Exosome PD-L1 (similar to tumor PD-L1) can bind to PD-1 on T cells, induce T cell apoptosis, and inhibit T cell activation and proliferation [
TDEs, through their miRNAs proteins, DNAs, mRNAs lncRNAs, initiate the transformation of epithelial cells to mesenchymal cells. This transformation was due to the loss of epithelial E-cadherin expression, cell-cell adhesion and cell polarity, and gaining of vimentin expression [16].
The complex and heterogeneous microenvironment of both primary or metastatic tumor is comprised of a network of cellular and acellular constituents. The cellular compartment consists of tumor cells and assorted non-transformed cells, such as cancer-associated fibroblasts (CAFs), macrophages, and endothelial cells. The non-cellular part is formed by secreted factors and components of the ECM. The tumor microenvironment modulates tumor progression by providing inhibitory or stimulatory growth signals [17]. Thus pre-metastatic niche refers to the microenvironment, that is primed to allow tumor cells to colonize in and disseminate to distant sites. The main machineries of the premetastatic niche formation include tumor-derived secreted factors (TDSFs), EVs bone marrow-derived cells (BMDCs), suppressive immune cells and host stromal cells [4], and inflammation. Chronic inflammation is a driving force for tumor development and metastasis. Thus, the local inflammatory microenvironment is one of the essential factors for the pre-metastatic niche formation and driving force for metastasis.
Tumor development and metastasis are aided by chronic inflammation. As a result, one of the most important variables in the establishment of a pre-metastatic niche is the local inflammatory microenvironment. Tumor cells can be induced to create TDSFs such as vascular endothelial growth factor (VEGF), tumor necrosis factor alpha (TNF-α), transforming growth factor (TGF-β), and interleukin-2 by the local inflammatory microenvironment. These TDSFs then exert a paracrine effect on myeloid cells, initiating their migration to potential pre-metastatic niche formation sites [18]. Host stromal cells in the pre-metastatic niche may upregulate the expression of inflammatory factors in response to TDSF activation. The recruitment of BMDCs or immune cells to the pre-metastatic niche speeds up the release of inflammatory factors. Exo from tumors also transport inflammatory substances into the bloodstream, where they reach the pre-metastatic niche. In the pre-metastatic niche, an inflammatory milieu supportive to tumors is eventually generated [18].
In a study conducted by Hoshino, he showed that the proinflammatory cytokine s100 was upregulated up to four folds when Kupffer cells were treated with integrin intact Exo, as compared to those treated with integrin knocked out Exo. Hoshino speculated that the activation of Src, and its phosphorylation might be a causative pathway [19].
TDE and MV were also shown to modify fibroblasts in the tumor stroma. When normal human fibroblasts were exposed to oral squamous carcinoma derived MV [20] the fibroblasts were altered into a cancer phenotype. This switch to CAFs was largely mediated via metabolic reprogramming of the fibroblasts to aerobic glycolysis, with an increase in glucose uptake and lactate secretion. Some TDEs contain surface TGF-β along with betaglycan, which could trigger SMAD-dependent signaling and regulate the differentiation of fibroblasts to myofibroblasts [21]. This was further proved by co-culturing the generated CAF with cancer cells which led to enhanced cancer cell invasion and migration, creating a bidirectional cross-talk that favors tumor promotion and spread. The MVs-induced fibroblast activation and spreading seem to occur in the matrix milieu in the tumor periphery [22]. In prostate cancer, TDE were shown to induce the expression of RANKL and Metalloproteinases in CAFs, through miR-100, -21, and -139, further promoting its metastasis [23]. Hypoxia seems to stimulate prostate cancer cells to release protein-rich Exo which further induces activation of CAFs [24], promotes epithethelial mesenchymal transition (EMT), stemness, and angiogenesis by prostate cancer cells.
Additionally, TDE were also described as regulators of metabolism in the tumor microenvironment, for example, breast cancer tumors could suppress glucose uptake by lung fibroblasts, via secretion of Exo containing miR-122, increasing glucose availability and facilitating metastasis [25]. The cell-to-cell communication mediated by Exo is also affected by the genetic profile of the recipient fibroblasts. For example, fibroblasts lacking the BRCA1, a tumor suppressor gene, internalize larger amounts of serum-derived Exo when compared to BRCA1 containing fibroblasts [26]. Furthermore, these cells were found to undergo a malignant transformation when exposed to Exo derived from sera of cancer patients, implying that oncosuppressor genes can prevent exosome information from tumor cells from being integrated and thus shelter these cells from their pro-oncogenic signals [26].
Tumor MVs extravasate through the vessel wall in pancreatic cancer, reach the liver microcirculation and are picked up by perivascular macrophages to prime the liver metastatic niche in a CD36-dependent manner. Furthermore, tumor MVs produced from the B16F10 melanoma cell line was discovered to cause metastases in BALB/c mice, which are generally resistant to the B1610 tumor cell line [27]. TDEs also protect cancer cells from apoptosis by selectively effluxing apoptosis inducer proteins that are delivered by T cells or natural killer (NK) cells. TDEs also reduce the effects of therapy by preventing drug efflux or concealing the binding site of monoclonal antibodies, which could lead to the emergence of chemotherapy-resistant cell populations [28].
Exosome-derived programmed death receptor 1 (PD-1) and programmed death-ligand 1 have been linked to an immunological escape mechanism in recent years. PD-1 is mostly found on macrophages, activated T cells, and B cells, whereas PD-L1 is abundant in tumor tissues, antigen-presenting cells (APCs), and stromal cells [29]. T lymphocytes can recognize and destroy tumor cells in normal circumstances. When PD-1 attaches to PD-L1, however, it sends an inhibitory signal to T cells, causing them to die and inhibiting their activation and proliferation. As a result, blocking the PD-1/PD-L1 pathway may boost the immune response by increasing the killing effect of T cells [30]. T lymphocytes can recognize and destroy tumor cells in normal circumstances. When PD-1 attaches to PD-L1, however, it sends an inhibitory signal to T cells, causing them to die and inhibiting their activation and proliferation (Figure 1). As a result, blocking the PD-1/PD-L1 pathway may boost the immune response by increasing the killing effect of T cells. As a result, Exo containing PD-L1 suppress the immune system in the pre-metastatic milieu and promote the establishment of a pre-metastatic niche [31].
Angiogenesis within the primary tumor is also influenced by tumor MVs and TDE. Normal endothelial cells (ECs) were shown to endocytose tumor EVs, which triggered PI3K/Akt signaling and increased EC motility and tube formation ability [32]. Tumor MVs and TDE also release VEGF, a pro-angiogenic substance that stimulates ECs [33]. Similarly, MVs produced from multiple myeloma cells have been demonstrated to transfer CD138, a myeloma cell marker, to ECs, promoting their proliferation, invasion, and production of the angiogenic mediators IL-6 and VEGF, resulting in tube formation [50] (Figure 2). MVs change the environment around the main tumor and create pre-metastatic niches from afar. This was originally attributed to their procoagulant activity, which encouraged the production of microthrombi and facilitated the extravasation of trapped circulating tumor cells. ECs are important components of the tumor microenvironment because they provide a pathway for nutrients and trophic substances [34].
Possible mechanisms of pre-metastatic niche formation. The figure delineates how TDEs can modulate its surroundings of ECM, cancer-CAFs, immune cells, ECs, and MSCs all in favor of tumor support and progression. TDE can carry integrins to distant sites and create a pre-metastatic niche.
TDE enriched in vascular cell adhesion molecule (VCAM)-1 and intercellular adhesion molecule (ICAM)-1 has been demonstrated to regulate the process of neovascularization in myeloid leukemia [35]. Furthermore, enhanced vascularization has been linked to the packaging of miR-92a in Exo derived from leukemia[36] and CO-029/D6.1A Tetraspanin in Exo produced from pancreatic cancer [37]. Upregulation of Heparanase in tumor cells, such as myeloma and breast malignancies, has also been linked to increased exosome production and exosomal packing of Syndecan-1, VEGF, and hepatocyte growth factor, resulting in enhanced endothelial invasion through the ECM [38]. Exo produced from skin cancer can also enhance angiogenesis by transferring the EGFR [39] and miR-9 to ECs [26]. Furthermore, melanoma-derived Exo have been found to condition sentinel lymph nodes prior to the installation of melanoma cells and subsequent metastasis by upregulating Collagen 18 and Laminin 5, as well as producing angiogenic growth factors [26].
Another significant component in altering tumor-EC communication is hypoxia. Hypoxic glioblastoma cells, for example, release Exo that interact with ECs, promoting proliferation and angiogenesis both in vitro and in vivo [40], and also prompting tissue factor/Factor VIIa dependent activation of hypoxic ECs [26].
Exo from melanoma cause pulmonary vascular leakiness and upregulate tumor cell recruitment genes such Stabilin 1, Vitronectin, Integrins, and Ephrin receptor b4 in lymph nodes, forming pre-metastatic niches [41]. Furthermore, breast cancer-derived Exo enriched in miR-105 alter the expression of Claudin 5, Zonula Occludens protein 1, and Occludin, which promotes metastasis by disrupting vascular endothelial barriers [42]. Exo produced from brain tumors include miR-181c, which regulates EC actin dynamics and promotes the breakdown of the blood-brain barrier by three times. Protein Kinase-1 Degradation Requires Phosphoinositol [43]. Similarly, glioblastoma cells release Exo with high quantities of VEGF-A, which promote EC permeability and angiogenesis in vitro [44].
TDE can promote pro-tumorigenic microenvironments via promoting tumor-stem/progenitor cell contact, in addition to its well-known actions in differentiated cells. Melanoma-derived Exo, for example, stimulate BMDCs by transferring the oncoprotein MET, resulting in the mobilization of vasculogenic and hematopoietic bone marrow progenitor cells to ensure vascular proliferation and immunosuppression at pre-metastatic niches [45]. Communication between tumor stem/progenitor cells is also critical in bone metastasis. Exo from bone metastatic prostate cancer PC3 cells were found to influence the process of bone metastasis by modulating both osteoclast genesis and osteoblast proliferation. Exo generated from osteoblasts, on the other hand, have been demonstrated to stimulate PC3 prostate cancer cell proliferation [46].
TDE was also demonstrated to influence the development of myeloid precursor cells into myeloid-derived suppressor cells (MDSCs), which are known to aid tumor progression by permitting immune escape [47]. Exo produced from breast carcinomas have been found to be taken up by bone marrow cells and to convert these cells’ development pathways toward MDSCs via Prostaglandin E2 and TGF-β, boosting COX2, IL6, VEGF, and Arginase1 accumulation by MDSCs [48].
TDE can also cause alterations in mesenchymal stem cells (MSCs), which help to promote and maintain tumor-promoting inflammatory environments. For example, HSP70+ lung tumor-derived exosomes (TDEs) activate NF-kB and cause MSCs to secrete IL-6, IL-8, and MCP1 via TLR2-mediated signaling, causing MSCs to become more inflammatory and tumor supportive [49]. According to De Veirman et al. [50], myeloma-derived Exo transfer miR-146a to mesenchymal cells, stimulating them to secrete numerous cytokines and chemokines including CXCL1, IL6, IL-8, IP-10, MCP-1, and CCL-5 (Figure 2). Another example is Exo produced by KMBC cholangiocarcinoma cells, which cause MSCs to upregulate IL-6, and hence KMBC cell proliferation [51].
One of the proposed mechanisms of tumorigenicity of TDE is the induction of tumor cell proliferation. Studies involving various cancer cells such as, chronic myeloid leukemia and in human gastric cancer, showed that this proliferative potential is via an autocrine induction through the phosphatidylinositol 3-kinase/protein kinase B (PI3K/AKT) and MAPK/ERK signaling pathways. Additionally, through the transference of lncRNAs (reviewed in [49]).
In addition, glioblastoma-derived Exo were shown to induce proliferation of the human glioma U87 cell line [40] in a mechanism dependent on the chloride intracellular channel protein 1 (CLIC1) [52]. In a more specific context linked to prostate cancer treatment, prostate cancer LNCaP cells grown in the presence of androgens generate Exo high in CD9, which enhance the growth of androgen-depleted LNCaP cells. Another example involves the promotion of in vivo growth of murine melanomas by systemic treatment of mice with melanoma-derived Exo, which accelerated growth and inhibited apoptosis of melanoma tumors in vivo [26].
TDE can alter the migratory behavior of recipient malignant cells. Exo produced from nasopharyngeal cancer-bearing EMT-inducing signals such as TGF-β and hypoxia-inducible factor 1 alpha (HIF1a) [53], matrix metalloproteinases (MMPs) Notch1, LMP1 Casein Kinase II and Annexin A2, were shown to enhance the migratory capacity of the tumor recipient cells. Another example involves Exo derived from hypoxic prostate cancer cells, which prompted invasiveness and motility of naïve human prostate cancer cells (reviewed in [26]) through the neighboring stroma and to nearby cells.
Exo have been found to have a role in tumor-tumor communication by transferring chemoresistance. Exo have been linked to the transfer of Docetaxel resistance in prostate cancer since Corcoran and colleagues first discovered it [54]. The transfer of cisplatin resistance in lung cancer is achieved by donor resistant cells producing Exo with low levels of miR-100-5p, which leads to enhanced expression of the mammalian target of rapamycin (mTOR) protein and chemoresistance in recipient cells [55].
MiRNA packed in Exo from drug-resistant cells can modulate the expression of specific target genes in breast cancer, such as miR-23a targeting Sprouty2, miR-222 targeting PTEN, miR-452 targeting APC4, and miR-24 targeting p27, thereby modulating chemoresistance in recipient cells that integrate these Exo. In fact, exosomal miR-222 plays a key role in this process, as the silencing of miR-221/222 prevents the transmission of resistance [56].
In addition to miRNAs, the transfer of exosomal mRNAs that encode drug-resistant proteins may result in chemoresistance in the receiving cell. GSTP1 exosomal mRNA from Adriamycin-resistant breast cancer cells, for example, confers resistance to previously susceptible cells. The presence of GSTP1 in circulating Exo from patients’ peripheral blood was linked to a worse outcome in breast cancer patients receiving Adriamycin [57]. A supporting stroma is required for an optimum metabolic and physiological environment for tumor growth. Fibroblasts are the most abundant cells in most solid tissues, participating in environmental cue responses and being a common target of tumor-derived signals [58].
Integrins are a wide family of cell adhesion receptor proteins such as alpha3beta1, alpha6beta1, alpha6beta4, and alpha7beta1. Their roles have been implicated in tumor metastasis and mesenchymal transformation. TDE carry these integrins from primary tumor sites to distant sites such as lung, lymph nodes, brain, and bone creating pre-metastatic niches (Figure 2) [59].
TDEs are involved in the advancement of several forms of cancer. Because of their abundance, TDEs may serve as noninvasive diagnostic and prognostic tools for various cancers. Additionally, blocking exosome secretion can slow the growth of some malignancies. Hence, Exo have been a popular target for developing cancer treatment techniques because of this property. Decreasing the expression of the exosomal proteins, Rab27a and Rab27b, inhibit exosome secretion without matching changes in soluble proteins secretions [60]. Several drugs used in the pharmaceutical industry such as Ketoconazole (an anti-fungal) sphingomyelinase (a hydrolase enzyme that is responsible for degrading sphingomyelin) [61], are additionally Rab27a inhibitors. These drugs can be re-directed as cancer modulators for their possible effects on attenuating TDE tumor progressive effects.
Furthermore, TDE owing to its small size, cancer-homing, and nontoxic nature, TDE can be re-directed to serve as a drug delivery system. Exo have been proven in several investigations to act as drug delivery vehicles, transporting anti-cancer chemicals to target cells [62]. For example, adriamycin and paclitaxel, target cancer cells via exosomal encapsulation and have low toxicity and immunogenicity [63].
EVs modulate the environment that favors tumor growth and progression. EVs provide a method of cell-cell communication, and through their rich cargo of ECM proteins, cell adhesion proteins, tyrosine kinases, chaperones, signaling proteins, DNA and RNA binding proteins, they create a pre-metastatic niche. By priming nearby and distant cells into becoming cancerous, they promote tumor metastasis. Several mechanisms have been discovered for their actions including, promotion of migratory behavior, chemoresistance, anti-apoptosis, vascular leakage, and immune modulation. Understanding how TDE and MVs create a pre-metastatic niche and how halting the trafficking of such vesicles can produce a revolutionizing new era in the field of cancer therapeutics. By preventing TDE-promoted metastasis and tumor progression, coupled with conventional radio and chemotherapy, the survival rates of cancer patients can significantly improve.
The author declares no conflicts of interest.
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Data deluge brings substantial challenges in the collection and management of massive amounts of unstructured data towards decision making. Likewise, unprecedented production of information exceeds the ability of authoritative bodies to create regulations and policies that can keep up with these transformations in the nature of work. We explicate the impact of well-timed policies (fiscal and monetary), prediction of long-term structural changes in the industrial sector, industrial strategy formulation practices, and examine the economic studies and analysis of sustainable development in these areas.",book:{id:"11198",title:"Digital Transformation",coverURL:"https://cdn.intechopen.com/books/images_new/11198.jpg"},signatures:"Kinda R. Dahlan, Ahmed A. 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The knowledge presented in this chapter may provide tips and inspiration for some other game projects, and practical and useful notes on the advantages or disadvantages of some systems will be interesting and useful.",book:{id:"11192",title:"Computer Game Development",coverURL:"https://cdn.intechopen.com/books/images_new/11192.jpg"},signatures:"Branislav Sobota, Marián Hudák and Emília Pietriková"},{id:"82300",title:"An Effective Method for Secure Data Delivery in IoT",slug:"an-effective-method-for-secure-data-delivery-in-iot",totalDownloads:2,totalDimensionsCites:0,doi:"10.5772/intechopen.104663",abstract:"The Internet of Things (IoT) has become very popular recently due to its important features that contribute to many aspects of our lives such as health and transportation. It consists of a vast number of different projects such as sensors, tags, actuators, and mobile devices, which can communicate and collaborate without human interactions. These devices carry small memory and low-energy battery, which affects their performance and lead to many issues. In this work, we are going to focus on the efficiency and security issues. We will propose a secure and efficient routing protocol for data delivery in order to improve its performance. The proposed technique will be evaluated in an implemented platform with appropriate case study. The expected outcome of this study will be a reference design and its practical implementation to support efficiency and security in IoT.",book:{id:"11197",title:"Internet of Things - New Trends, Challenges and Hurdles",coverURL:"https://cdn.intechopen.com/books/images_new/11197.jpg"},signatures:"Mnar Alnaghes, Nickolas Falkner and Hong Shen"},{id:"82267",title:"Methods for Speech Signal Structuring and Extracting Features",slug:"methods-for-speech-signal-structuring-and-extracting-features",totalDownloads:4,totalDimensionsCites:0,doi:"10.5772/intechopen.104634",abstract:"The preliminary stage of the biometric identification is speech signal structuring and extracting features. For calculation of the fundamental tone are considered and in number investigated the following methods – autocorrelation function (ACF) method, average magnitude difference function (AMDF) method, simplified inverse filter transformation (SIFT) method, method on a basis a wavelet analysis, method based on the cepstral analysis, harmonic product spectrum (HPS) method. For speech signal extracting features are considered and in number investigated the following methods – the digital bandpass filters bank; spectral analysis; homomorphic processing; linear predictive coding. This methods make it possible to extract linear prediction coefficients (LPC), reflection coefficients (RC), linear prediction cepstral coefficients (LPCC), log area ratio (LAR) coefficients, mel-frequency cepstral coefficients (MFCC), barkfrequency cepstral coefficients (BFCC), perceptual linear prediction coefficients (PLPC), perceptual reflection coefficients (PRC), perceptual linear prediction cepstral coefficients (PLPCC), perceptual log area ratio (PLAR) coefficients, reconsidered perceptual linear prediction coefficients (RPLPC), reconsidered perceptual reflection coefficients (RPRC), reconsidered perceptual linear prediction cepstral coefficients (RPLPCC), reconsidered perceptual log area ratio (RPLAR) coefficients. The largest probability of identification (equal 0.98) and the smallest number of coefficients (4 coefficients) are provided by coding of a vocal of the speech sound from the TIMIT based on PRC.",book:{id:"10992",title:"Speech Recognition - New Perspectives",coverURL:"https://cdn.intechopen.com/books/images_new/10992.jpg"},signatures:"Eugene Fedorov, Tetyana Utkina and Tetiana Neskorodieva"},{id:"82196",title:"Multi-Features Assisted Age Invariant Face Recognition and Retrieval Using CNN with Scale Invariant Heat Kernel Signature",slug:"multi-features-assisted-age-invariant-face-recognition-and-retrieval-using-cnn-with-scale-invariant-",totalDownloads:6,totalDimensionsCites:0,doi:"10.5772/intechopen.104944",abstract:"Face recognition across aging emerges as a significant area among researchers due to its applications such as law enforcement, security. However, matching human faces with different age gaps is still bottleneck due to face appearance variations caused by aging process. In regard to mitigate such inconsistency, this chapter offers five sequential processes that are Image Quality Evaluation (IQE), Preprocessing, Pose Normalization, Feature Extraction and Fusion, and Feature Recognition and Retrieval. Primarily, our method performs IQE process in order to evaluate the quality of image and thus increases the performance of our Age Invariant Face Recognition (AIFR). In preprocessing, we carried out two processes that are Illumination Normalization and Noise Removal that have resulted in high accuracy in face recognition. Feature extraction adopts two descriptors such as Convolutional Neural Network (CNN) and Scale Invariant Heat Kernel Signature (SIHKS). CNN extracts texture feature, and SIHKS extracts shape and demographic features. These features plays vital role in improving accuracy of AIFR and retrieval. Feature fusion is established using Canonical Correlation Analysis (CCA) algorithm. Our work utilizes Support Vector Machine (SVM) to recognize and retrieve images. We implement these processes in FG-NET database using MATLAB2017b tool. At last, we validate performance of our work using seven performance metrics that are Accuracy, Recall, Rank-1 Score, Precision, F-Score, Recognition rate and computation time.",book:{id:"11442",title:"Pattern Recognition - New Insights",coverURL:"https://cdn.intechopen.com/books/images_new/11442.jpg"},signatures:"Kamarajugadda Kishore Kumar and Movva Pavani"}],onlineFirstChaptersTotal:101},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:8,limit:8,total:0},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:89,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:104,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:31,numberOfPublishedChapters:314,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:11,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:141,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:129,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:113,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:105,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:18,numberOfOpenTopics:2,numberOfUpcomingTopics:1,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:5,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:14,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. The whole process of submitting an article and editing of the submitted article goes extremely smooth and fast, the number of reads and downloads of chapters is high, and the contributions are also frequently cited.",author:{id:"55578",name:"Antonio",surname:"Jurado-Navas",institutionString:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRisIQAS/Profile_Picture_1626166543950",slug:"antonio-jurado-navas",institution:{id:"720",name:"University of Malaga",country:{id:null,name:"Spain"}}}},{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}}]},series:{item:{id:"10",title:"Physiology",doi:"10.5772/intechopen.72796",issn:"2631-8261",scope:"Modern physiology requires a comprehensive understanding of the integration of tissues and organs throughout the mammalian body, including the cooperation between structure and function at the cellular and molecular levels governed by gene and protein expression. While a daunting task, learning is facilitated by identifying common and effective signaling pathways mediated by a variety of factors employed by nature to preserve and sustain homeostatic life. \r\nAs a leading example, the cellular interaction between intracellular concentration of Ca+2 increases, and changes in plasma membrane potential is integral for coordinating blood flow, governing the exocytosis of neurotransmitters, and modulating gene expression and cell effector secretory functions. 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His primary area of interest is physiology and pathophysiology of the gastrointestinal (GI) tract, with the major focus on the mechanism of GI mucosal defense, protection, and ulcer healing. He was a postdoctoral NIH fellow at the University of California and the Gastroenterology VA Medical Center, Irvine, Long Beach, CA, USA, and at the Gastroenterology Clinics Erlangen-Nuremberg and Munster in Germany. He has published 290 original articles in some of the most prestigious scientific journals and seven book chapters on the pathophysiology of the GI tract, gastroprotection, ulcer healing, drug therapy of peptic ulcers, hormonal regulation of the gut, and inflammatory bowel disease.",institutionString:null,institution:{name:"Jagiellonian University",institutionURL:null,country:{name:"Poland"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:5,paginationItems:[{id:"4",title:"Fungal Infectious Diseases",coverUrl:"https://cdn.intechopen.com/series_topics/covers/4.jpg",editor:{id:"174134",title:"Dr.",name:"Yuping",middleName:null,surname:"Ran",slug:"yuping-ran",fullName:"Yuping Ran",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bS9d6QAC/Profile_Picture_1630330675373",biography:"Dr. Yuping Ran, Professor, Department of Dermatology, West China Hospital, Sichuan University, Chengdu, China. Completed the Course Medical Mycology, the Centraalbureau voor Schimmelcultures (CBS), Fungal Biodiversity Centre, Netherlands (2006). International Union of Microbiological Societies (IUMS) Fellow, and International Emerging Infectious Diseases (IEID) Fellow, Centers for Diseases Control and Prevention (CDC), Atlanta, USA. Diploma of Dermatological Scientist, Japanese Society for Investigative Dermatology. Ph.D. of Juntendo University, Japan. Bachelor’s and Master’s degree, Medicine, West China University of Medical Sciences. Chair of Sichuan Medical Association Dermatology Committee. General Secretary of The 19th Annual Meeting of Chinese Society of Dermatology and the Asia Pacific Society for Medical Mycology (2013). In charge of the Annual Medical Mycology Course over 20-years authorized by National Continue Medical Education Committee of China. Member of the board of directors of the Asia-Pacific Society for Medical Mycology (APSMM). Associate editor of Mycopathologia. Vice-chief of the editorial board of Chinses Journal of Mycology, China. 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He joined the Department of Microbiology the same year and has been giving lectures on topics covering parasitology, immunology, molecular biology and industrial microbiology. He is currently a rated researcher by the National Research Foundation of South Africa at category C2. He has published widely in the field of infectious diseases and has overseen several MSc’s and PhDs. His research activities mostly cover topics on infectious diseases from epidemiology to control. His particular interest lies in the study of intestinal protozoan parasites and opportunistic infections among HIV patients as well as the potential impact of childhood diarrhoea on growth and child development. He also conducts research on water-borne diseases and water quality and is involved in the evaluation of point-of-use water treatment technologies using silver and copper nanoparticles in collaboration with the University of Virginia, USA. 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Dr. Saxena is a fellow of various international societies/academies including the Royal College of Pathologists, United Kingdom; Royal Society of Medicine, London; Royal Society of Biology, United Kingdom; Royal Society of Chemistry, London; and Academy of Translational Medicine Professionals, Austria. He was named a Global Leader in Science by The Scientist. 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