The effect of different growth medium and gelling agents on shoot regeneration capacity of flax hypocotyl explants
\r\n\r\n
\r\n\r\nVitamin K2 - Vital for Health and Wellbeing has been produced and distributed through the support from Kappa Bioscience, Norway.\r\n',isbn:"978-953-51-3020-8",printIsbn:"978-953-51-3019-2",pdfIsbn:"978-953-51-4895-1",doi:"10.5772/61430",price:139,priceEur:155,priceUsd:179,slug:"vitamin-k2-vital-for-health-and-wellbeing",numberOfPages:338,isOpenForSubmission:!1,isInWos:1,hash:"b2f9f024939ddc4f5da2a8afa3fcd9c9",bookSignature:"Jan Oxholm Gordeladze",publishedDate:"March 22nd 2017",coverURL:"https://cdn.intechopen.com/books/images_new/5169.jpg",numberOfDownloads:25964,numberOfWosCitations:7,numberOfCrossrefCitations:13,numberOfDimensionsCitations:26,hasAltmetrics:1,numberOfTotalCitations:46,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 6th 2015",dateEndSecondStepPublish:"January 31st 2016",dateEndThirdStepPublish:"March 25th 2016",dateEndFourthStepPublish:"May 30th 2016",dateEndFifthStepPublish:"August 31st 2016",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,editors:[{id:"36345",title:"Prof.",name:"Jan",middleName:"Oxholm",surname:"Gordeladze",slug:"jan-gordeladze",fullName:"Jan Gordeladze",profilePictureURL:"https://mts.intechopen.com/storage/users/36345/images/3823_n.jpg",biography:"Dr. Jan O. Gordeladze, Ph.D. (born 25th of April, 1950), holds a triple professor competence (Medical Biochemistry, Physiology, and Pharmacology), and is presently working as a Professor Emeritus at the Department of Biochemistry, Institute of Basic Medical Science, University of Oslo, Norway. He has previously been employed as the Medical Director of MSD, Norway, serving two years as a Fulbright scholar at the NIH, Bethesda, Maryland, USA. From 2006-2009 he was employed as Associate Professor at the University of Montpellier, France. He is a member of the Norwegian Stem Cell Center, and his research has over the past 7-10 years been devoted to differentiation of osteochondral cells from stem cells focusing on the impact of transcription factors and microRNA species constituting regulatory loop interactions with functional target genes. He has published more than 120 scientific articles, reviews/book chapters and presented more than 250 abstracts/posters/talks at conferences worldwide. Dr. Gordeladze has served as a Fulbright Scholar at The National Institute of Health, Bethesda, Washington DC during the years 1990-91.",institutionString:null,position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"9",totalChapterViews:"0",totalEditedBooks:"3",institution:null}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"379",title:"Vitaminology",slug:"alimentology-vitaminology"}],chapters:[{id:"53889",title:"Introductory Chapter: Vitamin K2",doi:"10.5772/66384",slug:"introductory-chapter-vitamin-k2",totalDownloads:1297,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Jan Oxholm Gordeladze",downloadPdfUrl:"/chapter/pdf-download/53889",previewPdfUrl:"/chapter/pdf-preview/53889",authors:[{id:"36345",title:"Prof.",name:"Jan",surname:"Gordeladze",slug:"jan-gordeladze",fullName:"Jan Gordeladze"}],corrections:null},{id:"51057",title:"Vitamin K, SXR, and GGCX",doi:"10.5772/63983",slug:"vitamin-k-sxr-and-ggcx",totalDownloads:1237,totalCrossrefCites:0,totalDimensionsCites:1,signatures:"Kotaro Azuma and Satoshi Inoue",downloadPdfUrl:"/chapter/pdf-download/51057",previewPdfUrl:"/chapter/pdf-preview/51057",authors:[{id:"184194",title:"Dr.",name:"Satoshi",surname:"Inoue",slug:"satoshi-inoue",fullName:"Satoshi Inoue"},{id:"189851",title:"Dr.",name:"Kotaro",surname:"Azuma",slug:"kotaro-azuma",fullName:"Kotaro Azuma"}],corrections:null},{id:"51024",title:"Vitamin K2 Rich Food Products",doi:"10.5772/63902",slug:"vitamin-k2-rich-food-products",totalDownloads:1496,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Muhammad Yasin, Masood Sadiq Butt and Aurang Zeb",downloadPdfUrl:"/chapter/pdf-download/51024",previewPdfUrl:"/chapter/pdf-preview/51024",authors:[{id:"178785",title:"Dr.",name:"Muhammad",surname:"Yasin",slug:"muhammad-yasin",fullName:"Muhammad Yasin"},{id:"182656",title:"Prof.",name:"Masood Sadiq",surname:"Butt",slug:"masood-sadiq-butt",fullName:"Masood Sadiq Butt"},{id:"189674",title:"Dr.",name:"Aurang",surname:"Zeb",slug:"aurang-zeb",fullName:"Aurang Zeb"}],corrections:null},{id:"50921",title:"Menaquinones, Bacteria, and Foods: Vitamin K2 in the Diet",doi:"10.5772/63712",slug:"menaquinones-bacteria-and-foods-vitamin-k2-in-the-diet",totalDownloads:2630,totalCrossrefCites:6,totalDimensionsCites:12,signatures:"Barbara Walther and Magali Chollet",downloadPdfUrl:"/chapter/pdf-download/50921",previewPdfUrl:"/chapter/pdf-preview/50921",authors:[{id:"184784",title:"Dr.",name:"Barbara",surname:"Walther",slug:"barbara-walther",fullName:"Barbara Walther"},{id:"188194",title:"Mrs.",name:"Magali",surname:"Chollet",slug:"magali-chollet",fullName:"Magali Chollet"}],corrections:null},{id:"54263",title:"The Impact of Vitamin K2 on Energy Metabolism",doi:"10.5772/67152",slug:"the-impact-of-vitamin-k2-on-energy-metabolism",totalDownloads:1344,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Mona Møller, Serena Tonstad, Tone Bathen and Jan Oxholm\nGordeladze",downloadPdfUrl:"/chapter/pdf-download/54263",previewPdfUrl:"/chapter/pdf-preview/54263",authors:[{id:"184157",title:"M.Sc.",name:"Mona",surname:"Møller",slug:"mona-moller",fullName:"Mona Møller"}],corrections:null},{id:"52078",title:"Vitamin K2 and Bone Health",doi:"10.5772/64876",slug:"vitamin-k2-and-bone-health",totalDownloads:1570,totalCrossrefCites:1,totalDimensionsCites:1,signatures:"Niels Erik Frandsen and Jan Oxholm Gordeladze",downloadPdfUrl:"/chapter/pdf-download/52078",previewPdfUrl:"/chapter/pdf-preview/52078",authors:[{id:"186115",title:"M.D.",name:"Niels Erik",surname:"Frandsen",slug:"niels-erik-frandsen",fullName:"Niels Erik Frandsen"}],corrections:null},{id:"54291",title:"Vitamin K2 and its Impact on Tooth Epigenetics",doi:"10.5772/66383",slug:"vitamin-k2-and-its-impact-on-tooth-epigenetics",totalDownloads:1623,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Jan Oxholm Gordeladze, Maria A. Landin, Gaute Floer Johnsen,\nHåvard Jostein Haugen and Harald Osmundsen",downloadPdfUrl:"/chapter/pdf-download/54291",previewPdfUrl:"/chapter/pdf-preview/54291",authors:[{id:"36345",title:"Prof.",name:"Jan",surname:"Gordeladze",slug:"jan-gordeladze",fullName:"Jan Gordeladze"}],corrections:null},{id:"51205",title:"Anti-Inflammatory Actions of Vitamin K",doi:"10.5772/63891",slug:"anti-inflammatory-actions-of-vitamin-k",totalDownloads:1724,totalCrossrefCites:2,totalDimensionsCites:4,signatures:"Stephen J. Hodges, Andrew A. Pitsillides, Lars M. Ytrebø and Robin\nSoper",downloadPdfUrl:"/chapter/pdf-download/51205",previewPdfUrl:"/chapter/pdf-preview/51205",authors:[{id:"184368",title:"Dr.",name:"Stephen",surname:"Hodges",slug:"stephen-hodges",fullName:"Stephen Hodges"},{id:"184370",title:"Dr.",name:"Robin",surname:"Soper",slug:"robin-soper",fullName:"Robin Soper"},{id:"184371",title:"Prof.",name:"Andrew",surname:"Pitsillides",slug:"andrew-pitsillides",fullName:"Andrew Pitsillides"}],corrections:null},{id:"50916",title:"Vitamin K2: Implications for Cardiovascular Health in the Context of Plant-Based Diets, with Applications for Prostate Health",doi:"10.5772/63413",slug:"vitamin-k2-implications-for-cardiovascular-health-in-the-context-of-plant-based-diets-with-applicati",totalDownloads:3108,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Michael S. Donaldson",downloadPdfUrl:"/chapter/pdf-download/50916",previewPdfUrl:"/chapter/pdf-preview/50916",authors:[{id:"180516",title:"Dr.",name:"Michael",surname:"Donaldson",slug:"michael-donaldson",fullName:"Michael Donaldson"}],corrections:null},{id:"50958",title:"Menaquinone‐4 Enhances Steroidogenesis in Testis Derived Tumor Cells Via the Elevation of cAMP Level",doi:"10.5772/63982",slug:"menaquinone-4-enhances-steroidogenesis-in-testis-derived-tumor-cells-via-the-elevation-of-camp-level",totalDownloads:1146,totalCrossrefCites:0,totalDimensionsCites:1,signatures:"Hsin‐Jung Ho, Hitoshi Shirakawa and Michio Komai",downloadPdfUrl:"/chapter/pdf-download/50958",previewPdfUrl:"/chapter/pdf-preview/50958",authors:[{id:"180389",title:"Dr.",name:"Hitoshi",surname:"Shirakawa",slug:"hitoshi-shirakawa",fullName:"Hitoshi Shirakawa"},{id:"180489",title:"MSc.",name:"Hsin-Jung",surname:"Ho",slug:"hsin-jung-ho",fullName:"Hsin-Jung Ho"},{id:"180490",title:"Prof.",name:"Michio",surname:"Komai",slug:"michio-komai",fullName:"Michio Komai"}],corrections:null},{id:"54241",title:"Vitamin K2 Facilitating Inter-Organ Cross-Talk",doi:"10.5772/67153",slug:"vitamin-k2-facilitating-inter-organ-cross-talk",totalDownloads:1364,totalCrossrefCites:1,totalDimensionsCites:1,signatures:"Jan O. Gordeladze, Håvard J. Haugen, Gaute Floer Johnsen and\nMona Møller",downloadPdfUrl:"/chapter/pdf-download/54241",previewPdfUrl:"/chapter/pdf-preview/54241",authors:[{id:"36345",title:"Prof.",name:"Jan",surname:"Gordeladze",slug:"jan-gordeladze",fullName:"Jan Gordeladze"}],corrections:null},{id:"51126",title:"Vitamin K2 in Animal Health: An Overview",doi:"10.5772/63901",slug:"vitamin-k2-in-animal-health-an-overview",totalDownloads:1333,totalCrossrefCites:0,totalDimensionsCites:1,signatures:"Jayde O’Neil, Bethany Scarrott, Ragnhild Aven Svalheim, Jonathan\nElliott and Stephen J. Hodges",downloadPdfUrl:"/chapter/pdf-download/51126",previewPdfUrl:"/chapter/pdf-preview/51126",authors:[{id:"184368",title:"Dr.",name:"Stephen",surname:"Hodges",slug:"stephen-hodges",fullName:"Stephen Hodges"},{id:"184369",title:"Ms.",name:"Jayde",surname:"O'Neil",slug:"jayde-o'neil",fullName:"Jayde O'Neil"},{id:"184748",title:"Ms.",name:"Bethany",surname:"Scarrott",slug:"bethany-scarrott",fullName:"Bethany Scarrott"},{id:"184749",title:"Prof.",name:"Jonathan",surname:"Elliott",slug:"jonathan-elliott",fullName:"Jonathan Elliott"},{id:"184750",title:"Ms.",name:"Ragnhild",surname:"Svalheim",slug:"ragnhild-svalheim",fullName:"Ragnhild Svalheim"}],corrections:null},{id:"50754",title:"Medicinal Chemistry of Vitamin K Derivatives and Metabolites",doi:"10.5772/63511",slug:"medicinal-chemistry-of-vitamin-k-derivatives-and-metabolites",totalDownloads:1188,totalCrossrefCites:2,totalDimensionsCites:2,signatures:"Shinya Fujii and Hiroyuki Kagechika",downloadPdfUrl:"/chapter/pdf-download/50754",previewPdfUrl:"/chapter/pdf-preview/50754",authors:[{id:"180528",title:"Prof.",name:"Hiroyuki",surname:"Kagechika",slug:"hiroyuki-kagechika",fullName:"Hiroyuki Kagechika"},{id:"180529",title:"Dr.",name:"Shinya",surname:"Fujii",slug:"shinya-fujii",fullName:"Shinya Fujii"}],corrections:null},{id:"50681",title:"From Protein Folding to Blood Coagulation: Menaquinone as a Metabolic Link between Bacteria and Mammals",doi:"10.5772/63342",slug:"from-protein-folding-to-blood-coagulation-menaquinone-as-a-metabolic-link-between-bacteria-and-mamma",totalDownloads:1265,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Brian M. Meehan and Jonathan Beckwith",downloadPdfUrl:"/chapter/pdf-download/50681",previewPdfUrl:"/chapter/pdf-preview/50681",authors:[{id:"180269",title:"Dr.",name:"Brian",surname:"Meehan",slug:"brian-meehan",fullName:"Brian Meehan"},{id:"185054",title:"Prof.",name:"Jon",surname:"Beckwith",slug:"jon-beckwith",fullName:"Jon Beckwith"}],corrections:null},{id:"52618",title:"Vitamin K2 Biosynthesis: Drug Targets for New Antibacterials",doi:"10.5772/65487",slug:"vitamin-k2-biosynthesis-drug-targets-for-new-antibacterials",totalDownloads:1433,totalCrossrefCites:0,totalDimensionsCites:2,signatures:"Michio Kurosu",downloadPdfUrl:"/chapter/pdf-download/52618",previewPdfUrl:"/chapter/pdf-preview/52618",authors:[{id:"33087",title:"Prof.",name:"Michio",surname:"Kurosu",slug:"michio-kurosu",fullName:"Michio Kurosu"}],corrections:null},{id:"50717",title:"Toxicological and Pharmacological Effects of VKOR Inhibitors",doi:"10.5772/63512",slug:"toxicological-and-pharmacological-effects-of-vkor-inhibitors",totalDownloads:1191,totalCrossrefCites:0,totalDimensionsCites:0,signatures:"Yohei Miyamoto",downloadPdfUrl:"/chapter/pdf-download/50717",previewPdfUrl:"/chapter/pdf-preview/50717",authors:[{id:"172105",title:"Ph.D.",name:"Yohei",surname:"Miyamoto",slug:"yohei-miyamoto",fullName:"Yohei Miyamoto"}],corrections:null},{id:"50994",title:"Enhanced Intracellular Delivery and Improved Antitumor Efficacy of Menaquinone-4",doi:"10.5772/63343",slug:"enhanced-intracellular-delivery-and-improved-antitumor-efficacy-of-menaquinone-4",totalDownloads:1028,totalCrossrefCites:1,totalDimensionsCites:1,signatures:"Kazuhisa Matsunaga, Munechika Enjoji, Yoshiharu Karube and Jiro\nTakata",downloadPdfUrl:"/chapter/pdf-download/50994",previewPdfUrl:"/chapter/pdf-preview/50994",authors:[{id:"179737",title:"Associate Prof.",name:"Kazuhisa",surname:"Matsunaga",slug:"kazuhisa-matsunaga",fullName:"Kazuhisa Matsunaga"},{id:"179741",title:"Prof.",name:"Munechika",surname:"Enjoji",slug:"munechika-enjoji",fullName:"Munechika Enjoji"},{id:"179742",title:"Prof.",name:"Yoshiharu",surname:"Karube",slug:"yoshiharu-karube",fullName:"Yoshiharu Karube"},{id:"179744",title:"Prof.",name:"Jiro",surname:"Takata",slug:"jiro-takata",fullName:"Jiro Takata"}],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},relatedBooks:[{type:"book",id:"5483",title:"Adiposity",subtitle:"Epidemiology and Treatment Modalities",isOpenForSubmission:!1,hash:"5f19b6a0755b8a29538e3b2043d4a854",slug:"adiposity-epidemiology-and-treatment-modalities",bookSignature:"Jan Oxholm Gordeladze",coverURL:"https://cdn.intechopen.com/books/images_new/5483.jpg",editedByType:"Edited by",editors:[{id:"36345",title:"Prof.",name:"Jan",surname:"Gordeladze",slug:"jan-gordeladze",fullName:"Jan Gordeladze"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6039",title:"Adiposity",subtitle:"Omics and Molecular Understanding",isOpenForSubmission:!1,hash:"7ef4705430dd255e9105eff55b3b21a8",slug:"adiposity-omics-and-molecular-understanding",bookSignature:"Jan Oxholm Gordeladze",coverURL:"https://cdn.intechopen.com/books/images_new/6039.jpg",editedByType:"Edited by",editors:[{id:"36345",title:"Prof.",name:"Jan",surname:"Gordeladze",slug:"jan-gordeladze",fullName:"Jan Gordeladze"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"5940",title:"Vitamin C",subtitle:null,isOpenForSubmission:!1,hash:"e23e79359167bb9d4a53edd78c7b5038",slug:"vitamin-c",bookSignature:"Amal H. Hamza",coverURL:"https://cdn.intechopen.com/books/images_new/5940.jpg",editedByType:"Edited by",editors:[{id:"188326",title:"Associate Prof.",name:"Amal",surname:"Hamza",slug:"amal-hamza",fullName:"Amal Hamza"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7263",title:"Vitamin E in Health and Disease",subtitle:null,isOpenForSubmission:!1,hash:"6bd8e547b4f3ad7f1675a36b8dbde8f2",slug:"vitamin-e-in-health-and-disease",bookSignature:"Jose Antonio Morales-Gonzalez",coverURL:"https://cdn.intechopen.com/books/images_new/7263.jpg",editedByType:"Edited by",editors:[{id:"109774",title:"Dr.",name:"Jose Antonio",surname:"Morales-Gonzalez",slug:"jose-antonio-morales-gonzalez",fullName:"Jose Antonio Morales-Gonzalez"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7038",title:"Vitamin D Deficiency",subtitle:null,isOpenForSubmission:!1,hash:"ba24f0913341357b0779ff9529c4bbfc",slug:"vitamin-d-deficiency",bookSignature:"Julia Fedotova",coverURL:"https://cdn.intechopen.com/books/images_new/7038.jpg",editedByType:"Edited by",editors:[{id:"269070",title:"Prof.",name:"Julia",surname:"Fedotova",slug:"julia-fedotova",fullName:"Julia Fedotova"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8594",title:"Fads and Facts about Vitamin D",subtitle:null,isOpenForSubmission:!1,hash:"1731029867f0d79c633e3408fc03ebd2",slug:"fads-and-facts-about-vitamin-d",bookSignature:"Edward T. Zawada Jr.",coverURL:"https://cdn.intechopen.com/books/images_new/8594.jpg",editedByType:"Edited by",editors:[{id:"16344",title:"Dr.",name:"Edward T.",surname:"Zawada Jr.",slug:"edward-t.-zawada-jr.",fullName:"Edward T. Zawada Jr."}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7978",title:"Vitamin A",subtitle:null,isOpenForSubmission:!1,hash:"dad04a658ab9e3d851d23705980a688b",slug:"vitamin-a",bookSignature:"Leila Queiroz Zepka, Veridiana Vera de Rosso and Eduardo Jacob-Lopes",coverURL:"https://cdn.intechopen.com/books/images_new/7978.jpg",editedByType:"Edited by",editors:[{id:"261969",title:"Dr.",name:"Leila",surname:"Queiroz Zepka",slug:"leila-queiroz-zepka",fullName:"Leila Queiroz Zepka"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7033",title:"Vitamin C",subtitle:"an Update on Current Uses and Functions",isOpenForSubmission:!1,hash:"719a5742e3271393fe43864e13e996cd",slug:"vitamin-c-an-update-on-current-uses-and-functions",bookSignature:"Jean Guy LeBlanc",coverURL:"https://cdn.intechopen.com/books/images_new/7033.jpg",editedByType:"Edited by",editors:[{id:"67023",title:"Dr.",name:"Jean Guy",surname:"LeBlanc",slug:"jean-guy-leblanc",fullName:"Jean Guy LeBlanc"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6709",title:"B Group Vitamins",subtitle:"Current Uses and Perspectives",isOpenForSubmission:!1,hash:"f34959a0fcc33a2c6fb3d03e9ec544bf",slug:"b-group-vitamins-current-uses-and-perspectives",bookSignature:"Jean Guy LeBlanc and Graciela Savoy de Giori",coverURL:"https://cdn.intechopen.com/books/images_new/6709.jpg",editedByType:"Edited by",editors:[{id:"67023",title:"Dr.",name:"Jean Guy",surname:"LeBlanc",slug:"jean-guy-leblanc",fullName:"Jean Guy LeBlanc"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,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:"Theophanides",surname:"Theophile",slug:"theophanides-theophile",fullName:"Theophanides Theophile"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],ofsBooks:[]},correction:{item:{id:"65668",slug:"corrigendum-to-clinical-applications-of-mesenchymal-stromal-cells-mscs-in-orthopedic-diseases",title:"Corrigendum to: Clinical Applications of Mesenchymal Stromal Cells (MSCs) in Orthopedic Diseases",doi:null,correctionPDFUrl:"https://cdn.intechopen.com/pdfs/65668.pdf",downloadPdfUrl:"/chapter/pdf-download/65668",previewPdfUrl:"/chapter/pdf-preview/65668",totalDownloads:null,totalCrossrefCites:null,bibtexUrl:"/chapter/bibtex/65668",risUrl:"/chapter/ris/65668",chapter:{id:"61187",slug:"clinical-applications-of-mesenchymal-stromal-cells-mscs-in-orthopedic-diseases",signatures:"Jiazhao Yang, Shiyuan Fang, Lei Xu, Li Li, Kai Xie, Jinsen Lu, Hao\nWang, Xujin Wang and Lixin Kan",dateSubmitted:"December 5th 2017",dateReviewed:"March 29th 2018",datePrePublished:"November 5th 2018",datePublished:"January 23rd 2019",book:{id:"6658",title:"Stromal Cells",subtitle:"Structure, Function, and Therapeutic Implications",fullTitle:"Stromal Cells - Structure, Function, and Therapeutic Implications",slug:"stromal-cells-structure-function-and-therapeutic-implications",publishedDate:"January 23rd 2019",bookSignature:"Mani T. Valarmathi",coverURL:"https://cdn.intechopen.com/books/images_new/6658.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"69697",title:"Dr.",name:"Mani T.",middleName:null,surname:"Valarmathi",slug:"mani-t.-valarmathi",fullName:"Mani T. Valarmathi"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:null}},chapter:{id:"61187",slug:"clinical-applications-of-mesenchymal-stromal-cells-mscs-in-orthopedic-diseases",signatures:"Jiazhao Yang, Shiyuan Fang, Lei Xu, Li Li, Kai Xie, Jinsen Lu, Hao\nWang, Xujin Wang and Lixin Kan",dateSubmitted:"December 5th 2017",dateReviewed:"March 29th 2018",datePrePublished:"November 5th 2018",datePublished:"January 23rd 2019",book:{id:"6658",title:"Stromal Cells",subtitle:"Structure, Function, and Therapeutic Implications",fullTitle:"Stromal Cells - Structure, Function, and Therapeutic Implications",slug:"stromal-cells-structure-function-and-therapeutic-implications",publishedDate:"January 23rd 2019",bookSignature:"Mani T. Valarmathi",coverURL:"https://cdn.intechopen.com/books/images_new/6658.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"69697",title:"Dr.",name:"Mani T.",middleName:null,surname:"Valarmathi",slug:"mani-t.-valarmathi",fullName:"Mani T. Valarmathi"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:null},book:{id:"6658",title:"Stromal Cells",subtitle:"Structure, Function, and Therapeutic Implications",fullTitle:"Stromal Cells - Structure, Function, and Therapeutic Implications",slug:"stromal-cells-structure-function-and-therapeutic-implications",publishedDate:"January 23rd 2019",bookSignature:"Mani T. Valarmathi",coverURL:"https://cdn.intechopen.com/books/images_new/6658.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"69697",title:"Dr.",name:"Mani T.",middleName:null,surname:"Valarmathi",slug:"mani-t.-valarmathi",fullName:"Mani T. Valarmathi"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}}},ofsBook:{item:{type:"book",id:"10818",leadTitle:null,title:"21st Century Approaches to Management and Accounting Research",subtitle:null,reviewType:"peer-reviewed",abstract:"
\r\n\tInterdisciplinary research necessitates an integration of disciplines in a collaborative approach. Accounting is an important decision-making tool for managers in every kind of business decision making therefore the accounting discipline can not be in isolation from the business itself. Specifically contemporary topics in strategic management and accounting such as corporate governance, innovation, inter-organizational relationships, and internationalization and their links to accounting in the form of blockchain technologies, earnings management, accounting quality, and ethics. Interdisciplinary research in accounting and strategic management, necessarily challenges each of these disciplines to deploy their own theories, analytical skills, and tools. The outcome of such efforts offers a holistic perspective on the business as a whole offering insights not only for researchers but also for practitioners.
\r\n\r\n\tOverall, the topics that are covered in this book will open up new avenues of research for scholars both in accounting and strategic management literature and offer unique insights to practitioners and decision-makers. This book is a must-read for everyone interested in novel perspectives in business research and practice.
\r\n\t
Plant tissue culture is a term containing techniques used to propagate plants vegetatively by using small parts of living tissues (explants) on artificial growth mediums under sterile conditions. Explants regenerate shoots and roots, and consequently whole fertile plants under certain cultural conditions. Micropropagation is the production of whole plants through tissue culture from small parts such as shoot and root tips, leaf tissues, anthers, nodes, meristems and embryos. Micropropagation is the vegetative (asexual) propagation of plants under in vitro conditions and is widely used for commercial purposes worldwide [1-3].
Plant tissue culture techniques have certain advantages over traditional ones of propagation. These are:
Thousands of mature plants can be produced in a short time that allows fast propagation of new cultivars,
Endangered species can be cloned safely,
Large quantities of genetically identical plants can be produced,
Plant production is possible in the absence of seeds,
The production of plants having desirable traits such as good flowers, fruits and odor is possible,
Whole plants can be regenerated from genetically modified plant cells,
Disease-, pest- and pathogen-free plants in sterile vessels are produced,
Plants that their seeds have germination and growing problems such as orchids and nepenthes, can be easily produced,
Providing infection-free plants for mass production is possible.
Plant tissue culture is based on totipotency which means that a whole plant can be regenerated from a single cell on a growth medium. One of the main objectives of tissue culture studies is to obtain high-frequency shoot regeneration, which is also a prerequisite for an efficient transformation system and a clonal propagation of plants with attractive flowers and fruits in large scale for ornamental purposes. Specially the introduction of foreign genes coding agronomically important traits into plant cells has no meaning unless transgenic plants are regenerated from the genetically modified cell(s).
It is known that some families and genera such as Solanacea (Nicotiana, Petunia and Datura), Cruciferae (Brassica and Arabidopsis), Gesneriaceae (Achimenes and Streptocarpus), Asteraceae (Chichorium and Chrysanthemum) and Liliaceae (Lilium and Allium) have a high regeneration capacity while regeneration in some other families such as Malvaceae (Gossypium) and Chenopodiaceae (Beta) is difficult. In order to increase the regeneration capacity of explant from the genotype of interest, we have to find answers to such questions as “Why do some genotypes regenerate easily?”, “What can be done to increase the regeneration capacity of explant?”.
Plant material is extremely important for the success of tissue culture studies [4]. Factors affecting explant’s tissue culture response are (1) genotype, (2) physiological stage of donor plant, (3) explant source, (4) explant age, (5) explant size, (6) explant position in donor plant and (7) explant density. Plant segments used in tissue culture as explant are stem [5], root [6], leaf [7], flower [8], ovule [9], cotyledon and hypocotyl [10, 11]. Such these explants form direct and indirect organs and embriyos. Thin cell layer can also be used as explant in some species [12] while embryos can be successfully used in cereals [13]. Moreover, shoot tips and meristems may give successful results for callus formation and shoot regeneration [14].
Regeneration capacity of plants shows a wide range among families, species and even within genotypes from the same species (Figure 1). Generally dicotyledons regenerate more easily than monocots. Plants from some dicotyledon families such as Solanacea, Cruciferae, Gesneriaceae, Begoniaceae and Crassulaceae have a high regeneration capacity. In general, herbaceous plants regenerate more easily than woody plants such as trees and shrubs [3].
Sugarbeet from Chenopodiaceae family is known as a recalcitrant genotype with respect to in vitro culture and genetic transformation [15, 16] (Figure 2) while regeneration and transformation are quite easy in tobacco from Solanaceae family [17]. It was reported that somatic embryogenesis changed from 0.00% to 77.50% in 14 maize genotypes cultured in vitro [18].
Adventitious shoot regeneration from hypocotyl explants of flax (Linum usitatissimum L.) cultivars a. \'1886 Sel.\' and b. \'Clarck\' 4 weeks after culture initiation
Shoot regeneration from petiole explants of sugar beet (Beta vulgaris L.) 4 weeks after culture initiation. Only three explants out of 10 regenerated.
Explants show an ability to express totipotency are the most suitable for tissue culture [19]. Generally, vegetative segments of plants regenerate more easily in vitro than generative ones [20]. Explants should be isolated from healthy plants with high cell division for successfull response to tissue culture. On the other hand, regeneration capacity of mature tissues is quite low. In general, young tissues and organs have a high regeneration capacity than the older ones. Regeneration capacity of genotypes increases during flowering period as in Lunaria annua [21], Ranunculus sceleratus [22]. Although plants in a resting stage (dormant) are generally difficult to culture in vitro [3], there are some exceptions as flower stem explants of Tulipa form shoots only in dry storage (dormant) stage [23].
Plants grown under greenhouse conditions give rise to better results than the ones grown in field conditions [24]. There are huge variations regarding tissue culture response in explants excised from plants grown in field condition depending on wheather conditions during the year [3]. However, the best results are obtained from explants excised from in vitro grown seedlings [25].
In a study conducted by Yildiz et al. [25], regeneration capacity of flax (Linum usitatissimum L.) hypocotyl explants isolated from in vitro- and greenhouse-grown seedlings was compared. Five mm sections of hypocotyl explants from in vitro-grown seedlings were directly cultured on MS medium supplemented with 1 mg l-1 BAP and 0.02 mg l-1 NAA for shoot regeneration while hypocotyls from greenhouse-grown seedlings were surface-sterilized before culture initiation. The highest values with respect to shoot regeneration percentage, shoot number per explant and total shoot number per petri dish were recorded in explants isolated from in vitro-grown seedlings of all cultivars.
87.50 shoots were formed over 100.00 in explants isolated from in vitro-grown seedlings while only 22.50 shoots were recoved in explants of greenhouse-grown seedlings in cv. \'1886 Sel.\'. The highest shoot number per explant and total shoot number per petri dish were obtained in explants isolated from in vitro-grown seedlings as 14.43 and 126.26, respectively. Specially, total shoot number per petri dish was 42.95 times more in explants of in vitro-grown seedlings as 126.26 than in explants excised from greenhouse-grown seedlings as 2.94. It was reported that neither shoot regeneration percentage nor shoot number per explant is lonely an indicator of the success of tissue culture studies but \'total shoot number per petri dish\' is a good indicator of the success in both parameters for the genotype of interest [26]. These figures in all the parameters indicated the significance of explant source very well.
Regeneration capacity of older plants is often low. As the organ using for explant source gets older, regeneration capacity decreases. An example of differences in regeneration capacity between young and old seedlings that are used as source of explant was flax (Linum usitatissimum L.) [27]. In this study, shoot regeneration capacity of hypocotyl explants excised from in vitro-grown seedlings at different ages (7, 12 and 17 days) was examined in three flax cultivars. Explants of 7-day-old seedlings gave rise to the highest results with respect to shoot regeneration percentage, shoot number per explant and total shoot number per petri dish. The explants of 7-day-old seedlings were reported to be more vital and well grown (Figure 3).
Shoot regeneration from flax (Linum usitatissimum L.) cv. \'1886 Sel.\' hypocotyl explants excised from a. 7-day-old seedling and b. 12-day-old seedling 4 weeks after culture initiation
In the study, it was observed that shoot regeneration percentage, shoot number per explant and total shoot number per petri dish varied excessively among explants from different seedling ages. All these three parameters, which were the highest in explants of 7-day-old seedling were reduced significantly in explants of 12- and 17-day-old seedlings. Results clearly showed that there were statistically significant differences in all parameters examined in all cultivars among explants at different ages.
It is so difficult to obtain a successful tissue culture response from small parts such as cells and meristems than from larger parts such as leaves and hypocotyls due to their limited nutrients and hormone reserves. Larger explants having a big amount of nutrition reserves such as tubers and bulbs can easily regenerate in vitro and are less dependent on nutrients and hormones in growth medium [3].
In vitro growth of explants can be affected depending on the place from where they are excised. For instance, the higher parts of donor plant are older than the lower parts. Evers [28] has reported that shoot initials from lower parts of Pseudotsuga menziesii developed better under in vitro conditions. Explants excised from the base produce adventitious bulbs more easily than the top parts of the bulbs [3].
In a study conducted with flax, tissue culture response of hypocotyl explants at different positions was evaluated. Hypocotyls were classified from where they were excised as top, medium and low. Top part was just below the cotyledon leaves while lower one was close to bottom (Figure 4). Fifteen hypocotyls from 7-day-old in vitro-grown seedlings were cultured on MS medium 1 mg l-1 BAP and 0.02 mg l-1 NAA for 4 weeks.
Explants excised from top (just below cotyledon leaves) part of the seedling gave rise to the highest results with respect to shoot regeneration percentage, shoot number per explant and total shoot number per petri dish. Results were getting lower in the explants excised from medium part of the seedling. And the lowest values were recorded in the explants from lower part (Unpublished study results) (Figure 5).
The highest values in shoot regeneration percentage, shoot number per explant and total shoot number per petri dish were recorded in the explants excised from the top of the seedling as 97.78%, 5.17 and 76.00, respectively. These figures were the lowest in hypocotyl segments in the lower part of the seedling as 66.67%, 2.62 and 26.33, respectively. There were huge difference between the results of explants excised from the top and the low parts of the seedling. Shoot regeneration percentage was 97.78% in explants of the top part of the seedling while it was 66.67%. That is 97.78 explants regenerated over 100.00 in the top part of the seedling.
Mean shoot number per explant was recorded as 5.17 in explants excised from the top parts of the seedling while it was 2.62 in explants isolated from the low part of the seedling. Similar results were obtained in total shoot number per petri dish. Shoot number per explant and total shoot number per petri dish are the main indicators of success in plant tissue culture. Especially, after transformation studies via Agrobacterium tumefaciens, regeneration capacity of tissue decreases siginificantly due to plant defense mechanism against pathogen attact. That is why, the number of shoots regenerated should be as much as possible.
Seven-day-old flax (Linum usitatissimum L.) cv. \'Madaras\' seedling and the position from which explants were excised 1. top, 2. medium and 3. low. Bar = 1.0 cm
Tissue culture response of hypocotyl explants excised from different positions of flax (Linum usitatissimum L.) cv. \'Madaras\' seedling a. top, b. medium and c. low
There are many studies about competition among the plants in field conditions. Plant leaves compete for irradiance, and roots for water and nutrients [29, 30]. High plant density was accepted as a biotic stress factor [31, 32]. Stoffella and Bryan [33] have reported that plant density has an effect on plant development and yield of many vegetable crops. A linear increase has indicated in fruit yield when plant density is increased [34-36]. Abubaker [37] has noted that the highest planting density gave rise to the lowest yield in bean due to the high competition among plants for water and minerals. Asghari et al. [38] have reported that the chicory plant increased root diameter for increased absoption of water under high density by high competition between plants.
Since a plant is a unity of cells and tissues, behavior of cells and tissues as the smallest unit of the organism, represents the plant’s response against any factors arising from nearby environment. Competition and stress are the brother concepts that should be taken into account to increase the success of tissue culture studies. Explants under in vitro conditions compete with each other for a constant amount of water and nutrients in growth medium. It was revealed that competition was one of the most important factors increasing explant’s regeneration capacity [32].
Yildiz [32] has reported that encouraging hypocotyl explants of flax (Linum usitatissimum L.) for competition by decreasing the culture distances among them increased shoot regeneration capacity remarkably till a certain point from where stress initiated and significant decreases in all parameters were observed (Figure 6 and Figure 7). For shoot regeneration, hypocotyls were cultured in a petri dish (90 × 90 mm) at 0.5 x 0.5, 1.0 x 1.0, 1.5 x 1.5 and 2.0 x 2.0 cm distances.
Results clearly showed that there were statistically significant differences in fresh and dry weights in all cultivars among explants cultured at different culture distances. The highest fresh and dry weights per explant of all cultivars were obtained from 2.0 x 2.0 cm distance of hypocotyls and they decreased by decreasing distances. These findings were supported by Gersani et al. [39] and Maina et al. [40] who reported that plants grown alone produce more biomass or yield than those grown with the others. It could be concluded that the decreased distance in which explants were cultured induced stress caused likely by the deficiency of water, sucrose and nutrients. Increases in the fresh and dry weights were chiefly due to an increase in the absorption of water and other components from the basal medium [41]. It was stated that the fresh weight increase was mainly due to cell enlargement by water absorption [42] and increase in dry weight was closely related to cell division and new material synthesis [43].
The highest shoot number per hypocotyl (20.70 in \'Madaras\', 14.57 in \'1886 Sel.\' and 17.40 in \'Clarck\') and the highest shoot length (3.10, 2.14 and 2.09 cm, respectively) were obtained at 1.0 x 1.0 cm distance in all cultivars studied. It is thought that competition among explants cultured at 1.0 x 1.0 cm distance encouraged them to give higher results than at the other distances.
Plant growth regulators are perhaps the most important components affecting shoot regeneration capacity of explants [44]. In tissue culture studies, correct combinations of auxins and cytokinins have been tried to be determined for high frequency shoot regeneration for the explant [26]. This study has shown that determination of optimum levels of auxins and cytokinins in growth medium is not the only way of increasing shoot regeneration capacity, but also shoot regeneration frequency of explants could be increased simply by encouraging explants into competition.
Development of explants cultured at different distances 0.5 x 0.5 cm (A, a), 1.0 x 1.0 cm (B, b), 1.5 x 1.5 cm (C, c) and 2.0 x 2.0 cm (D, d); six weeks after culture initiation. Bar is 1.0 cm for petri dishes and 0.5 cm for shoot regeneration
Competition-stress curve of flax cultivars \'Madaras\', \'1886 Sel.\' and \'Clarck\' with respect to shoot number per hypocotyl and shoot length
All tissue culture studies which aim to obtain high-frequency shoot regeneration, which is also a prerequisite for an efficient transformation system, should be performed under sterile conditions. Explant health is the main factor determining regeneration capacity. Viability of explant and the seedling from which the explant is excised, are very important for high-frequency shoot regeneration [45]. The most important treatment prior to culture initiation is perhaps surface-sterilization of the explant. Since in vitro conditions provide bacteria and fungi with an optimal growth environment, unsuccessful sterilization hinders the progress of tissue culture studies. Surface-sterilization process aims to eliminate all microorganisms that can easily grow under in vitro conditions; on the other hand, it should guarantee the explant’s viability and regeneration capacity, which are known to be affected by the concentration, application period [46] and temperature [45] of disinfectant. Since direct contact of explant with disinfectant during the sterilization process may have a severe effect on regeneration capacity of the tissue [45], using aseptic tissues as source of explant is highly recommended [47, 48].
A wide range of surface disinfectants, such as ethanol, hydrogen peroxide, bromine water, mercuric chloride, silver nitrate, and antibiotics are used for surface-sterilization; however sodium hypochlorite (NaOCl) has been most widely used. NaOCl is highly effective against all kinds of bacteria, fungi, and viruses [49-52]. Moreover, NaOCl has a strong oxidizing property which makes it highly reactive with amino acids [53, 54], nucleic acids [55], amines, and amides [56, 57]. The general reaction between amino acids and NaOCl produces the respective aldehyde, NH4Cl and CO2 [54].
In vitro seed germination, seedling growth and the viability of the tissue were negatively affected by sodium hypochlorite (NaOCl) at high concentrations [45, 58, 59] while it is uneffective for sterilization of tissues at low concentrations. The negative effects of NaOCl concentration become more severe with increasing application period. Since regeneration capacity of the tissue is negatively affected by higher concentrations and longer application periods of disinfectants [3, 46], sterilization process under in vitro conditions should aim to use the lowest concentration of disinfectant for the shortest time.
In the study aiming to evaluate the effects of NaOCl solutions used for sterilization on in vitro seed germination and seedling growth in Lathyrus chrysanthus Boiss., the best results were obtained from 3.75% NaOCl concentration and 15 min. application period for all parameters examined [60]. Seedborne contamination increased gradually by decreasing concentrations and application periods of NaOCl below 3.75% and 15 min. Dramatic decreases were observed at 5.00% NaOCl concentration in all cases. At this concentration, NaOCl showed deleterious effect on the embryo of the seed. Seed germination decreased to 65.18% when NaOCl concentration increased to 5.00% from 3.75% at 15 min. application period. Seedling growth from seeds sterilized with 3.75% NaOCl concentration for 15 min. were observed growing faster than that of sterilized with other concentrations and application periods of NaOCl (Figure 8). By increasing NaOCl concentration to 5.00%, seedling length decrased to 3.45 cm from 3.90 cm. Higher results in seedlings grown from seeds sterilized with 3.75% NaOCl concentration for 15 min. could be caused by higher tissue water content as reported that in vitro explant growth and plantlet establishment have been affected significantly by tissue water content [41].
In vitro seedling growth from Lathyrus chrysanthus seeds sterilized with (a) 2.50%, (b) 3.75% and (c) 5.00% NaOCl for 15 min.
It was firstly reported that besides concentration and application period, temperature of NaOCl was also one of the most important factors affecting in vitro seed germination, seedling growth and explant’s regeneration capacity [45].
At the 2.00% NaOCl concentration using for surface-sterilization of flax (Linum usitatissimum L.) seeds, when the temperature of NaOCl was set below 10°C, bacterial and fungal contamination was observed. However, increases in NaOCl temperature above 10°C resulted in dramatic decreases in seed germination, seedling growth, hypocotyl and root lengths. When flax seeds were surface-sterilized with 3.00% and 4.00% NaOCl concentrations at 30°C, seed germination, seedling growth, hypocotyl and root lengths decreased dramatically. Decreases in all parameters in NaOCl temperature above 10°C could be the fact that disinfection activity of NaOCl increases [61] and disinfectant penetrates more easily through the seed coat [62]. Higher NaOCl temperatures resulted in morphologically abnormal seedlings with stunted hypocotyls and roots (Figure 9).
Yildiz and Er [45] reported that increasing disinfectant temperature using for surface-sterilization of flax seeds to obtain sterile in vitro seedlings from which hypocotyls were isolated, reduced shoot regeneration significantly.
In vitro seedling growth in flax from seeds sterilized with 2.00% NaOCl at temperatures of (a) 10ºC, (b) 20ºC and (c) 30ºC
Regeneration capacity of the explant is negatively affected by higher concentrations, application periods and temperatures of disinfectant used for surface-sterilization. In the sterilization process of the tissues, the concentration, application period and temperature of NaOCl solutions are closely related to each other and they should be considered together. Direct contact of the tissue with disinfectant during the sterilization process may have a severe effect on the viability and regeneration capacity depending on concentration, temperature and application periods of disinfectant [45, 46]. In addition to this common knowledge, not only seed germination and seedling growth are directly affected by sterilization process, but also regeneration capacity of explants and health of regenerated shoots are indirectly influenced significantly in plant tissue culture studies. That means NaOCl affects the success of in vitro studies, from in vitro seed germination and seedling establishment to regeneration capacity of the tissue and recovery of plantlets.
The composition of growth medium is an important factor affecting growth and morphogenesis of plant tissues. Plant tissue culture medium consists of macronutients, micronutrients, vitamins, amino acids or other nitrogen supplements, carbon sources, organic supplements, solidifying agents and growth regulators. Murashige and Skoog [63] are the most commonly used medium in plant tissue culture. The B5 [64], N6 [65] and Nitsch and Nitsch [66] (NN) have been widely used for many plant species. Moreover, for culture of woody species, the Driver/Kuniyuki walnut medium (DKW) [67] and the WPM medium [68] are used. The growth medium is selected for the purpose of tissue culture and for the plant species [69].
Yildiz et al. [27] have conducted a study to evaluate the effects of two different growth medium (MS and B5) and two gelling agents (Agar and Phytagel) on the regeneration capacity of flax (Linum usitatissimum L.) hypocotyl explants of three cultivars namely \'Madaras\', \'1886 Sel.\' and \'Clarck\'. Results showed that MS (Murashige and Skoog) growth medium and Agar as gelling agent gave rise to the highest results with respect to shoot regeneration percentage, shoot number per explant and total shoot number per petri dish in all cultivars studied (Table 1).
Cultivars | Shoot regeneration (%) | Shoot number per explant | Total shoot number per petri dish | |||||||||
1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | 1 | 2 | 3 | 4 | |
\'Madaras\' | 97.22 | 91.67 | 80.56 | 91.67 | 7.41a | 4.70b | 2.56cd | 2.50cd | 86.67a | 56.34bc | 30.67cde | 30.00cde |
\'1886 Sel.\' | 100 | 100 | 100 | 100 | 6.72a | 5.56ab | 4.14cd | 3.95cd | 80.34a | 66.67ab | 49.67cd | 47.34cd |
\'Clarck\' | 97.22 | 88.89 | 88.89 | 94.44 | 4.81a | 3.02b | 2.59bc | 2.34bc | 57.67a | 35.00b | 31.00bc | 28.00bcd |
The effect of different growth medium and gelling agents on shoot regeneration capacity of flax hypocotyl explants
After explants placed on growth medium for different purposes, they should be cultured in culture rooms where the environmental factors such as temperature and light are controlled. Different species may need different environmental conditions for successful culture.
Lighting in culture rooms is realized by fluorescent tubes. Control equipments of tubes should be set up outside the culture room. Otherwise they may cause over heating inside the room and in that case extra cooling is necessary. Due to light sources inside the culture room, there should be an efficient cooling system to maintain constant temperature conditions. Fluorescent tubes can be installed under the shelves, above the cultures which provide a more uniform irradiation for the cultures. Although 16h light and 8h dark photoperiod is usually used, there may be some differences according to species grown under long-day or short-day conditions.
The temperature in culture room is so important for successful tissue culture. Temperature variation in culture room should be as small as possible and generally ±1ºC is allowed. Otherwise, changing temperature regime causes stress in cultures which is one of the main reasons of unsuccess. That is why, working with many culture rooms are recommended instead of working only with one.
Water is the source of life on earth. Life in a large proportion of terrestrial ecosystems is limited by water availability. The water content of an actively growing plant can be as much as 95% of its live weight. A plant requires water as an essential ingredient of photolysis, the photochemical stage of photosynthesis where water is split using light energy. Neither carbon dioxide nor oxygen required for photosynthesis is usable by plant unless it is in solution in water. Therefore, water is the key to plant’s survival and growth. Water is also an excellent solvent. The substances (solutes) that become dissolved in water in plants include mineral ions such as potassium (K+), sugars (glucose and sucrose), and amino acids, main components of proteins.
The reduction in growth, yield and quality by water stress has been well recognized in field conditions [70-71]. Germination and seedling establishment guarantee plant survival and are very important phases of plant life. Germination rate decreases with decreasing external water potential and for each species there is a critical value of water potential below which germination will not occur [72].
Yildiz and Ozgen [41] have reported that tissue water content affected explant’s shoot regeneration capacity significantly. In the study, water-treated and non-water treated hypocotyl explants of three flax cultivars were compared with regard to fresh and dry weights, shoot regeneration percentage, shoot number per explant, shoot length and total shoot number per petri dish. Some hypocotyls were submerged in sterile distilled water with a gentle shaking for 20 min before placing on regeneration medium, while others were directly cultured on MS medium supplemented with 1 mg l-1 6-benzylaminopurine (BAP) and 0.02 mg l-1 naphthaleneacetic acid (NAA) for regeneration. Results clearly showed that there were sharp and statistically significant differences in all cultivars between water-treated and non-water-treated explants concerning all characters examined (Table 2, Figure 10).
Possibly, pretreatment of explants with water softened the epidermis layer and increased the permeability which caused to high tissue metabolic activity by increasing water and hormone uptake from the medium. Thus, increase in the fresh and dry weights of water-treated hypocotyl explants at the end of culture were chiefly due to an increase in the absorption of water and other components from the basal medium via high permeable epidermis membrane. In the study, non-water-treated explants were found smaller than water-treated ones in all cultivars. Dale [42] stated that the fresh weight increase is mainly due to cell enlargement by water absorption, cell vacuolation, and turgor-deriven wall expansion. The increase in dry weight was closely related to cell division and new material synthesis [43]. Dry weight increase of water-treated explants is due to an increase in carbohydrate metabolism resulting from increased water uptake. On the other hand, lower levels of all parameters of non-water-treated explants were directly due to a decreased water uptake from the environment and consequently, a reduced mobilization of plant growth regulators. Hsiao [73] has reported that the inhibition of growth under water stress conditions is the result of inhibition of cell division, cell elongation or both. Osmotic water absorption affects cell elongation. It has been suggested that osmotic stress modifies the biochemical changes taking place in the cell wall during growth thereby preventing extension [74]. The primary action of osmotic inhibition is retardation of water uptake which is vital for germination and growth [75]. It has been stated that water stress alters the level of plant hormones [76].
Cultivar | Hypocotyl | Shoot regeneration (%) | |||||
Fresh weight (g) | Dry weight (g) | ||||||
WT | NWT | WT | NWT | WT | NWT | ||
\'Madaras\' | 0.430±0.047 | 0.216±0.019 | 0.034±0.002 | 0.025±0.002 | 100±0.000 | 75.00±2.937 | |
\'1886 Sel.\' | 0.343±0.011 | 0.231±0.013 | 0.029±0.013 | 0.021±0.001 | 100±0.000 | 73.34±4.051 | |
\'Clarck\' | 0.396±0.013 | 0.192±0.025 | 0.031±0.001 | 0.019±0.002 | 100±0.000 | 78.33±2.305 | |
Cultivar | Shoot number per hypocotyl | Shoot length (cm) | Total shoot number per petri dish | ||||
WT | NWT | WT | NWT | WT | NWT | ||
\'Madaras\' | 12.17±0.210 | 8.00±0.328 | 0.56±0.043 | 0.25±0.018 | 182.50±3.142 | 119.88±4.90 | |
\'1886 Sel.\' | 11.20±0.114 | 8.15±0.367 | 0.58±0.029 | 0.22±0.009 | 167.88±1.712 | 122.25±5.487 | |
\'Clarck\' | 10.83±0.265 | 5.26±0.491 | 0.60±0.043 | 0.27±0.020 | 162.50±3.974 | 78.88±7.392 |
Adventitious shoot regeneration from water-treated (WT) and non-water-treated (NWT) hypocotyls of three flax cultivars 6 weeks after culture initiation on MS medium containing 1 mg l-1 BAP and 0.02 mg l-1 NAA.
Treatment of explants with water before culture initiation increased permeability of the epidermis layer and the tissue’s water content and so enabled water, all solutes and plant growth regulators to transfer into the tissue more easily, providing all cells with a high regeneration capacity and consequently increasing explant’s tissue culture response.
Shoots regenerated from water-treated and non-water-treated explants were rooted on MS medium containing 3 mg l-1 IBA for 3 weeks. The best results were obtained in the shoots regenerated from water-treated explants (Figure 10c). Sharp and dramatic differences, which were all statistically significant at the 0.01 level, were observed in all parameters between the shoots regenerated from water-treated and non water-treated explants (Table 3). Similar effects of water treatment were also noted in rooting stage. This means that shoots regenerated from water-treated explants were more capable of establishing new plantlets than the ones grown from non-water-treated explants.
In vitro shoot regeneration from water-treated (a) and non-water-treated (b) hypocotyls of Linum usitatissimum cv. \'1886 Sel.\' from 6-week-old culture. In vitro rooting and development of shoots regenerated from water-treated (c) and non-water-treated (d) hypocotyls of Linum usitatissimum cv. \'1886 Sel.\' 3 weeks later.
Cultivar | Shoot length (cm) | Number of roots | Mean length of each root (cm) | |||
WT | NWT | WT | NWT | WT | NWT | |
\'Madaras\' | 3.02±0.225 | 1.65±0.209 | 10.20±1.519 | 6.50±1.053 | 1.58±0.156 | 1.16±0.192 |
\'1886 Sel.\' | 3.98±0.220 | 1.28±0.185 | 21.31±2.121 | 7.19±1.342 | 1.92±0.144 | 0.82±0.076 |
\'Clarck\' | 4.81±0.396 | 2.10±0.156 | 29.00±2.887 | 14.63±1.812 | 2.33±0.223 | 1.56±0.143 |
In vitro root development of shoots regenerated from water-treated (WT) and non-water-treated (NWT) hypocotyl explants on rooting medium enriched with 3 mg l-1 IBA 3 weeks after culture initiation
In another study conducted by Yildiz et al. [77], pretreated and non-pretreated hypocotyl explants of three flax cultivars (\'Omega\', \'Fakel\' and \'Ariane\') were cultured for adventitious shoot regeneration. Two different pretreatment applications were compared to the conventional regeneration protocol with respect to hypocotyl fresh and dry weights, shoot regeneration percentage, shoot number per hypocotyl, shoot length and total chlorophyll content. In the 1st and 2nd pretreatment applications, hypocotyl explants were kept in sterile cabin under air flow for 30 min. in order to make them dry as reported by Christmann et al. [78] to decrease the tissue water content and to gain explants the ability of uptaking increased amount of water, all solutes and plant growth regulators by using osmotic pressure in consequent applications. Then explants were treated with MS solution containing 1 mg l-1 BAP and 0.02 mg l-1 NAA for 15 min. Finally all explants were cultured on MS medium without growth regulators in the 1st pretreatment application and on MS medium enriched with 1 mg l-1 BAP and 0.02 mg l-1 NAA in the 2nd pretreatment application. It was expected that by immersing explants into liquid regeneration medium after drying enabled all cells to absorb more growth regulators along with water in both pretreatment applications. However, only in 2nd pretreatment application, explants were cultured on MS medium containing 1 mg l-1 BAP and 0.02 mg l-1 NAA which means that tissues maintained uptaking increased water and growth regulators from the regeneration medium which led to higher results in all parameters studied as reported by Yildiz and Ozgen [41]. Likewise, Okubo et al. [79] have reported that endogenous hormone levels of tissue affected regeneration capacity in vitro significantly. Fatima et al. [80] have also noted that internal factors such as chemicals and mineral nutrients affect in vitro plant growth. The required amount of exogenous plant growth regulators for cultured tissues depends on the endogenous levels plant tissues have [80]. It was first reported that keeping explants in sterile distilled water for 20 min. before culture on MS medium enriched with 1 mg l-1 BAP and 0.02 mg l-1 NAA increased the regeneration capacity of hypocotyls of flax tremendously by increasing permeability of the epidermis layer and tissue’s water content and enabling water, all solutes and growth regulators to transfer into the tissue more easily [41].
According to the results, there were statistically significant differences among pretreated and non-pretreated hypocotyls in all cultivars (Table 4).
Pre. App. | Hypocotyl | Shoot regeneration (%)* | Shoot number per hypocotyl | Shoot length (cm) | Total chlorophyll content (µg/g fresh tissue) | ||
Cultivar | Fresh weight (g) | Dry weight (g) | |||||
\'Omega\' | 1 | 0.25**±0.020c | 0.014 ±0.0019b | 82.40±1.07b | 6.76±0.46b | 1.93±0.13b | 217.1±10.40c |
2 | 0.48±0.023a | 0.034 ±0.0023a | 100.00±0.00a | 11.38 ±0.69a | 2.82±0.14a | 380.6±26.91a | |
3 | 0.37±0.017b | 0.018 ±0.0017b | 90.00±5.77ab | 7.99±0.74b | 1.51±0.17b | 286.2±10.45b | |
\'Fakel\' | 1 | 0.21±0.027b | 0.016 ±0.0030b | 72.00 ±7.53b | 6.42±0.19c | 1.05±0.11b | 197.0±15.40c |
2 | 0.42±0.006a | 0.032 ±0.0046a | 100.00±0.00a | 8.89±0.37a | 1.61±0.18a | 316.5±14.37a | |
3 | 0.31±0.045b | 0.024 ±0.0031ab | 80.60±7.45a | 7.49±0.10b | 0.96±0.10b | 252.1±9.89b | |
\'Ariane\' | 1 | 0.19±0.024b | 0.014 ±0.0023b | 46.23 ±6.20c | 4.26±0.18b | 1.27±0.06b | 192.0±11.25c |
2 | 0.36±0.055a | 0.030 ±0.0012a | 100.00 ±0.00a | 6.64±0.25a | 2.00±0.13a | 346.0±18.62a | |
3 | 0.26±0.026ab | 0.019 ±0.0018b | 65.35 ±3.76b | 4.59±0.07b | 1.06±0.09b | 268.2±24.10b |
Tissue culture response from pretreated and non-pretreated hypocotyls of three flax cultivars 6 weeks after culture initiation
The highest results in both fresh and dry weights of hypocotyls of all cultivars were obtained from 2nd pretreatment application. Scores of fresh and dry weights were followed by non-pretreated hypocotyls. The lowest results were recorded from 1st pretreatment application in all cultivars studied (Table 4). From the results, it could be concluded that increases in the fresh and dry weights were chiefly due to an increase in the absorption of water and growth regulators from the medium where explants were first pretreated and then cultured. When the results of 2nd pretreatment application were examined, it could be easily seen that culturing explants on MS medium containing 1 mg l-1 BAP and 0.02 mg l-1 NAA after treating them with liquid MS medium supplemented with 1 mg l-1 BAP and 0.02 mg l-1 NAA clearly enriched the tissue’s growth regulators level which caused to higher fresh and dry weights.
The results related to shoot regeneration percentage indicated that the lowest results were obtained from the 1st pretreatment application in all cultivars. Hypocotyl explants formed roots and fewer callus in the 1st pretreatment application than the others. All explants regenerated successfully in the 2nd pretreatment application and consequently shoot regeneration percentage was recorded as 100% in all cultivars studied (Table 4, Figure 11).
The highest results in shoot number per hypocotyl and shoot length were obtained from 2nd pretreatment application in all cultivars studied. The highest shoot number per hypocotyl was recorded as 11.38 in \'Omega\', 8.89 in \'Fakel\' and 6.64 in \'Ariane\'. The highest scores related to shoot length were 2.82, 1.61 and 2.00 cm in \'Omega\', \'Fakel\' and \'Ariane\', respectively. Shoot regeneration capacity of hypocotyls increased significantly in 2nd pretreatment application. The best results in total chlorophyll content were obtained from 2nd pretreatment application in all cultivars. The highest scores of total chlorophyll content were recorded as 380.6 µg/g fresh tissue in \'Omega\', 316.5 µg/g fresh tissue in \'Fakel\' and 346.0 in \'Ariane\' (Table 4). The explants to which 2nd pretreatment application was carried out were more vital and well-grown and more capable of regeneration (Figure 11). Emerson [81] reported that there is a close relationship between photosynthesis and chlorophyll content. Chlorophyll content of leaf is considered as a sign of photosynthetic capacity of tissues [81-84] which plays a critical role in plant growth and development [85] and its amount changes under stress conditions [86-88]. Gireesh [89] has reported that chlorophyll can be used to measure growth.
In vitro shoot regeneration from pretreated and non-pretreated hypocotyls of flax (Linum usitatissimum) cv. \'Omega\'. (a) 1st pretreatment application: Hypocotyls were waited for 30 min. in sterile cabin under air flow and treated with solution containing 1 mg/l BAP and 0.02 mg/l NAA for 15 min. and finally cultured on MS0 medium, (b) 2nd pretreatment application: Hypocotyls were waited for 30 min. in sterile cabin under air flow and treated with solution containing 1 mg/l BAP and 0.02 mg/l NAA for 15 min. and finally cultured on MS medium containing 1 mg/l BAP+0.02 mg/l NAA, (c) Non-pretreatment application: Hypocotyls were directly cultured on MS medium containing 1 mg/l BAP+0.02 mg/l NAA
From the results, it could be concluded that the lower levels of all parameters recorded in the 1st and 3rd pretreatment applications were directly due to a decreased uptake of water and growth regulators from the medium. Tissue culture response has been affected significantly by tissue water content [41]. Treatment of explants with liquid MS medium containing 1 mg l-1 BAP and 0.02 mg l-1 NAA for a while before culture initiation enabled water, all solutes and plant growth regulators to transfer into the tissue much more, providing all cells with a high regeneration capacity and consequently increasing explant’s tissue culture response.
Plant tissue culture techniques help us to propagate plants vegetatively in a large amount starting from small parts of a tissue and by using the potential of known as totipotency, to form a whole, fertile plant. Plant tissue culture studies are performed on an artificial growth medium under sterile conditions. Explants regenerate shoots and roots, and consequently whole fertile plants under certain cultural conditions. Tissue culture studies aim to obtain high-frequency shoot regeneration, which is also a prerequisite for an efficient transformation system and a clonal propagation of plants. The introduction of foreign genes coding agronomically important traits into plant cells has no meaning unless transgenic plants are regenerated from the genetically modified cell(s). Therefore, using tissues having high regeneration capacity is extremely important. Regeneration capacity of cells or tissues to be used in transformation studies, affects the success of genetic transformation significantly. The types and concentrations of plant growth regulators in plant cell culture significantly affect growth and morphogenesis. In order to obtain high frequency adventitious shoot regeneration for related genotype, correct concentrations and combinations of auxins and cytokinins should be determined. However, determining the explant type, and correct concentrations and combinations of growth regulators is not enough for high frequency shoot regeneration. Since every cell has an ability of forming a whole fertile plant under in vitro conditions, shoot regeneration frequency can always be higher than we obtain in theory. Many factors affecting regeneration capacity of explant are not found out yet. For instance, a recently reported technique utilizing competition among explants is very effective to increase shoot regeneration capacity. Thus, unknown factors affecting regeneration capacity of explants should be determined in order to increase the success of tissue culture studies.
Tuberculosis (TB) continues to be a major public health concern with over 2 billion people currently infected, 8.6 million new cases per year, and more than 1.3 million deaths annually [1]. The current drug-regimen combination for drug sensitive TB consists of isoniazid, rifampicin, ethambutol and pyrazinamide, administered over 6 months [2]. If this treatment fails, second-line drugs are used, such as para-aminosalicylate (PAS) and fluoroquinolones, which are usually either less effective or more toxic with serious side effects. Although this regimen has a high success rate, it is marred by compliance issues, which have resulted in the rise of multidrug resistant (MDR), extensively drug resistant (XDR) and totally drug resistant (TDR) strains of the causative agent, Mycobacterium tuberculosis (Mtb) [3, 4], in both immunocompetent and immunocompromised patients worldwide [5]. However, it took about 40 years for a new TB drug to be discovered and most of the current TB drugs target a total of only ~10 proteins, even though the complete genome of Mtb was published nearly 20 years ago [6]. Consequently, most of the essential proteins are orphans since their ligands are still to be identified. In our context, target deorphaning or deconvolution encompasses identification of ligands for Mtb proteins not currently exploited in TB chemotherapy and those of old TB targets. Targeting further essential proteins should allow the fight against drug resistance to be enhanced, and possibly lead to a reduction in the duration of TB treatment.
The conventional target deorphaning process involves experimental work, which characteristically includes genetic, proteomics and transcriptional profiling and then identification of the ligands for the proteins using many more chemical-proteomic approaches [7]. This approach is usually long, expensive and time consuming. However, developments in bioinformatics and chemoinformatics, together with advances in computer tools and resources, have fortunately revolutionised target deorphaning. Bioinformatics describes the target space in Mtb from the genome to the proteome, whilst chemoinformatics provides information about the available chemical space and tools for navigation of the space. Together these developments have led to a mushrooming of computer-based target deorphaning methods ranging from modelling proteomes, virtual screening, machine and deep learning, and chemogenomics [8, 9, 10]. When used effectively in conjunction with experimental work, computational methods can facilitate identification of new TB targets and drugs [11, 12, 13].
Therefore, in this chapter we present an overview of the genome of Mtb, giving a detailed account on how the computational techniques have been used to de-orphan Mtb targets including case studies, the current and proposed future impacts of these techniques on the number of de-orphaned Mtb targets and their impacts in boosting the biomedical efficacy of TB drugs. The collated data will provide researchers in academia and industry with knowledge of target-ligand pairs and interactions, information crucial for the design of novel drugs with known targets that are less prone to resistance, with minimal side effects and interactions with e.g. anti-HIV drugs.
An extensive literature search was performed to give an overview of the genome of the Mtb and status of the currently used tuberculosis drugs and their targets. An analysis of the essential proteins in Mtb and the number of proteins targeted by the current TB drugs was performed. To boost this data Mtb target-ligand data was extracted from the ChEMBL database version 24 (
Cole and co-workers [14] in 1998 reported the complete sequence of Mtb, which comprises of 4,411,529 base pairs. The genome has an evenly distributed guanine-cysteine content of 65.6% and represents the second-largest bacterial genome sequence currently available. Additionally, the genome is rich in repetitive DNA, particularly insertion sequences, and in new multi-gene families and duplicated housekeeping genes, providing evidence for horizontally-transferred pathogenicity islands of a particular base composition [14].
The genome of Mtb has some exceptional features, for example there are over 200 genes that encode enzymes for the metabolism of fatty acids, comprising 6% of the total (Table 1). Among these, about 100 are predicted to function in the oxidation of fatty acids. This large number of Mtb enzymes that putatively have fatty acids as substrates may be linked to the ability of this pathogen to grow in the tissues of the infected host, where fatty acids maybe the major carbon source. Another unusual feature of the Mtb genome is the presence of the unrelated Pro-Glu (PE) and Pro-Pro-Glu (PPE) families of proteins that have conserved N-terminal domains of 100 and 180 amino acids respectively. The antigenicity of these proteins has led to the assumption that at least some of these proteins may be involved in antigenic variation of Mtb during infection [15].
Function | No. of genes | % of total genes | % of total coding capacity |
---|---|---|---|
Lipid metabolism | 225 | 5.7 | 9.3 |
Information pathways | 207 | 5.2 | 6.1 |
Cell wall and cell processes | 517 | 13.0 | 13.5 |
Stable RNAs | 50 | 1.3 | 0.2 |
IS elements and bacteriophages | 137 | 3.4 | 2.5 |
PE and PPE Proteins | 167 | 4.2 | 7.1 |
Intermediary metabolism and respiration | 877 | 22.0 | 24.6 |
Regulatory proteins | 188 | 4.7 | 4.0 |
Virulence, detoxification and adaptation | 91 | 2.3 | 2.4 |
Conserved hypothetical function | 911 | 22.9 | 18.4 |
Proteins of unknown function | 607 | 15.3 | 9.9 |
General classification of Mtb genes. Adopted from [15].
The success of TB chemotherapy derives from an “intensive” phase involving a cocktail of four first-line drugs, comprising, rifampicin (RIF), isoniazid (INH), pyrazinamide (PZA), and ethambutol (EMB). A threatening global issue of this epidemic is the emergence of drug-resistant bacteria, a trend that is on the rise, as such strains are easily spread with low fitness costs associated with transmission [16]
In addition, comorbidity with HIV causes massive diagnostic and therapeutic challenges and results in adverse drug interactions [19]. This is because RIF is a potent inducer of drug-metabolising enzymes, including cytochrome P450 (CYP) 3A4. This induction dramatically reduces plasma levels of several highly active antiretroviral therapy drugs; thus, patients are often forced to complete their TB treatment before beginning HIV treatment [20]. Patients who contract MDR-TB with HIV have a very poor prognosis due to the duration of treatment; these individuals frequently succumb within a few months. Therefore, there is an urgent need to develop continually new active agents to combat MDR-TB which has been compounded by the emergence of XDR-TB. Furthermore, cases of TDR-TB have been noted in China, India, Africa, and Eastern Europe. In TDR-TB, the Mycobacterium are resistant to all available therapeutics [19]. To address this, in 2012 the U.S. Food and Drug Agency (FDA) approved bedaquiline for MDR-TB [21] and later delamanid was approved as a compassionate care option for XDR-TB and TDR-TB infections, nonetheless the EMA approved both agents for MDR-TB [22]. The biggest challenge is that these drugs have reported human ether-a-go-go related gene (hERG) toxicity, as well as multiple absorption, distribution, metabolism and excretion (ADME) issues due to their high lipophilicity [21]. This leads to an urgent need for development of new agents that have successful therapeutic effects.
To date the number of essential Mtb proteins encoded by approximately 4000 genes is just over 500 (Figure 1), and this provides a rich source for novel targets for new and current TB drugs. However, Lamichhane et al. [23] reported that TB chemotherapy exploited only 10 of these proteins; Table 2, gives a summary of the targets, and their current and/or new drug ligands. The most popular target is enoyl[acyl-carrier protein] reductase, important for the biosynthesis of mycolic acid. Efforts to identify genes that code for new potential drugs are underway, as evidenced by 76 TB data points recorded in the ChEMBL database version 24 (
Circular diagram of the genome of Mtb genes, essential proteins and the number of proteins that are drug targets.
Targets | Function | Conventional drugs | New ligands |
---|---|---|---|
Enoyl-(acyl-carrier-protein) reductase (InhA), Fatty acid synthase | Biosynthesis of mycolic acids, that is essential for growth and virulence | Isoniazid Ethambutol Pyrazinamide Delamanid | Tetrahydropyrans (PT070) Methylthiazoles Diazaborines Pyrrolidine-carboxamide Piperazine indoleformamides Aminoproline Arylamides Imidazopiperidines |
DNA gyrase | An ATP-dependent enzyme that acts by creating a transient double-stranded DNA break | Fluoroquinolones | Clinafloxacin |
Ubiquinol-cytochrome C-reductase (QCrB) | Electron carriers of the respiratory chain | Pyrrolo[3,4-c]pyridine-1,3(2H)diones Lansoprazole | |
Transmembrane transport protein large (MmpL3) | Responsible for heme uptake into the cell. Responsible for the transport of ions, drugs, fatty acids and bile salts | SQ109 Adamantyl ureas Phenylpyrroles Benzimidazoles Tetrahydropyrazolo [1,5-a]pyrimidine-3-carboxamide Spiropiperidines | |
Decaprenylphospo-β-D-ribofuranose-2-oxidase (DprE1) | Cell wall synthesis | Benzothiazinones (BTZ043) Benzothiazole (TCA1) 4-aminoquinolone piperidine amides 2-carboxyquinoxalines Oxadiazoles Benzo [b]thiophenes Pyrazolopyridones | |
RNA polymerase | Responsible for transcription | Rifampicin Rifapentine Rifabutin | |
Protein synthase | Protein synthesis | Linezolid ( | PNU100480 AZD5847 |
ATP Synthase | ATP synthesis | Bedaquiline | D-Dethiobiotin |
Cytidine triphosphate (CTP) synthetase | Catalysis of amination of uridine triphosphate (UTP) into CTP | Thiophenecarboxamide 4-(pyridine 2-yl) thiazole | |
Transcription factor (IdeR) | Regulating the intracellular levels of iron | Benzo-thiazol benzene sulfonic acid | |
Lysine-ε-amino transferase (LAT) | Catalysing reversibly the transamination of lysine into α-ketoglutaric acid | Benzothiazole |
This is of paramount importance because Mtb secreted proteins play a vital role in host-pathogen interactions and facilitate nutrient acquisition, pilot the host immune response and interfere with therapeutic intervention. Therefore, the Mtb secretome consists of proteins essential for successful invasion and in vivo growth during host infection. The essential proteins are the most suitable drug targets for the development of diagnostic tools and new drugs, because of their key role in in vivo bacterial survival and growth. Identifying ligands for these proteins required for growth and survival in the infected host could lead to the discovery of potentially useful biomarkers to add on the above mentioned drug targets [27].
The development in genomics, coupled with advances in high performance computing and validation of molecular targets, has introduced new approaches to drug discovery that provide a shift from the historical pipeline that focuses on target identification and in most cases involves single targets. In this era of extensive discovery of new chemical entities for treatment of TB and other infectious diseases like HIV/AIDs, a number of research institutes as well as pharmaceutical companies are eagerly developing computational tools and protocols to facilitate drug discovery and development [28]. Genomics provide DNA, RNA, transcriptomic and proteomic data that is housed in a variety of databases and provide resources e.g. from the European Bioinformatics Institute (EBI)
The revolution in genomics led to the availability of a number of mycobacterial genomes and the development of a variety of databases consisting of Mtb genomic and transcriptomic data. The genomic databases provide information about the structure, function and evolution of Mtb genes, whilst the transcriptomics provide information crucial for analysis of gene expression using large scale RNA sequences [29]. On the other hand proteomics provides information about the function, networks and structure of proteins. In their paper, Machado et al. [29] give a detailed summary of most computational resources for TB and we encourage readers to consult the article for more information. Similarly a number of chemogenomic resources and database containing data for Mtb ligand annotated targets have been developed. Examples of such databases include the ChEMBL database [30], a database of small bioactive molecules and their targets, TIBLE [31] a database containing MIC and target data for mycobacterial species and TDR targets containing target-ligand information for neglected tropical diseases including TB. The databases are freely available and provide easy access to target-ligand data for Mtb. In these databases each target is associated to ligand(s) obtained from bioassays and vice versa.
A number of computational methods are being explored in order to identify ligands for both host and pathogen targets and for targets from other organisms like Plasmodium falciparum [32]. In most cases two or more complementary ligand-based and structure-based deorphaning approaches are used; statistical methods involving machine learning [8] and deep learning strategies are applied in conjunction with biological and/or biophysical methods to validate the computational results or the computational methods are used to provide the protein-ligand binding information in the absence of X-ray co-crystallised structures of the ligand [12, 13]. In their work, Mendes and Blundell [13] applied cheminformatics to complement current efforts for target identification of fragment-sized molecules that target e.g. the PanC that synthesises pantothenate important for generation of the Mtb co-enzyme A. This has led to the identification of ‘hotspots’ in the binding pockets of a number of proteins, which highlight the most favoured binding spots for the protein. Hotspots and druggability will be discussed in detail in Section 6.
Structure-based virtual screening is an approach used in drug discovery to computationally screen small molecule databases for compounds that target proteins of known 3D structure that are experimentally validated. Brain Shoichet [33] has pointed out that this approach was first published in the 1970s, however most new ligands and their targets were not identified until the early 2000. The method offers the opportunity to access a large number of potential new chemical ligands for old and new targets. In the presence of available ligands for named biological targets, ligand-based virtual screening may be used using a variety of techniques ranging from molecular similarity, pharmacophoric search, to machine learning and most recently deep learning.
Structure-based virtual screening plays a significant role in drug discovery in that it is used to identify ligands for biological targets when the 3D structures of the Mtb targets from X-ray crystallography, nuclear magnetic resonance (NMR) or cryoelectron microscopy are available in the Protein Data Bank, or homology models available in the CHOPIN database and/or generated in house. This method applies structural data of proteins/receptors to provide small molecules with specific structural attributes for good binding affinity [34]. Generally, the process involves three crucial steps, namely preparation of 3D crystal structures of proteins obtained from the Protein Data Bank (PDB) and the ligand structures, docking calculation and data analysis. Protein structure preparation involves adding hydrogen atoms that are normally missing in the coordinate files, adding missing residues, optimising hydrogen bonds, removing atomic clashes, as well as sampling the degrees of freedom such as flip that are not clear in standard resolution crystal structures, for example the 180o flips of chain terminal rotatable side-chain groups e.g. in shape-symmetric amino acids Asn and Gln, tautomer and/or ionisation state and relaxation of the target and ligand structure [35]. Most docking software is associated with protein and ligand preparation tools, for example Autodock4 or VINA require structures prepared using AutoDockTools (ADT) and the protein preparation script to generate Autodock-type atoms containing Gasteiger charges, and produce the pdbqt files that are compatible with the tool [36]. Similarly, the Primex and Ligprep tools are used to prepare the protein and ligand structures respectively before docking with GLIDE [37]. The quality of input structure files contribute to the quality of the docking results, and the importance of protein and ligand preparation have been highlighted by Sastry [35].
Molecular docking calculations are capable of predicting the binding conformation of ligands inside the binding pocket of a target, as such they are used to map small molecules onto targets and hence provide essential binding information for structure-based drug design. To achieve this, a number of docking algorithms like Autodock [36], perform a stochastic conformational search or e.g. in GLIDE, a [37] that perform a systematic search [34]. In a stochastic search structural parameters, such as torsional, translational and rotational degrees of freedom of the ligand, are randomly modified to generate an ensemble of molecular conformations and increase the chances of finding the energy global minimum, whilst in a systematic conformational search structural features are gradually changed until a local or global minimum is reached [34]. During the search, conformations of a number of potential binding compounds are explored and evaluated using a specific scoring function. In addition, the conformations are ranked based on their calculated binding energy. Highly ranked compounds are selected as ligands for the target. On the other hand, reverse or inverse docking is used for identifying targets of drug phenotypic hits from a sea of targets. In this way, structure-based screening helps to identify and explain polypharmacology, molecular mechanism of action of substances, facilitate drug repurposing, detect adverse drug reactions and hence toxicity.
In an effort to de-orphan the HTH transcription regulator, EthR, and identify the binding mode of the ligand, we docked 200 fragment-like compounds from the Maybridge database to the highest quality crystal structure of the 23 PDB entries using the GOLD algorithm (unpublished work). We used Arpeggio [38], an online tool that identifies non-covalent interactions in protein-structures, to assess the role of each EthR binding site residue and each small-molecule ligand moiety in contributing to protein-ligand interactions. Visual assessment of interactions involved calculating interactions using the Arpeggio web server (
(A) Binding modes of two fragment-like molecules inside the long cylindrical allosteric binding pocket of EthR defined by five helices. Yellow sticks depict the molecule occupying the upper binding site close to the entrance of the pocket and cyan sticks represent a molecule occupying the inner binding site close to the HTH domain. (B) EthR-ligand interactions involving Trp103 (yellow) at the entrance of the binding pocket of the protein. Ligand atoms and bonds are in pink, grey rings are hydrophobic interactions, red rings show hydrogen bonds. (C) EthR-ligand (pink) interactions involving Phe110 located at the center of the binding pocket of EthR.
Similarly, docking calculations were used to assess binding of ligands identified from for a novel TB drug target, inosine monophosphate dehydrogenase (IMPDH) protein Guab2 that is responsible for the synthesis of xanthosine monophosphate (XMP) from IMP, identified from high throughput screening [12]. Hit compounds were identified in a single shot high-throughput screen, validated by dose response and subjected to further biochemical analysis. The compounds were also assessed using molecular docking experiments, providing a platform for their further optimisation using medicinal chemistry. From the results, it was observed that occupation of the nicotinamide sub-site was correlated with interactions of the ligands with the purine ring of IMP.
Likewise, we used a combination of ligand-based and structure-based chemogenomic approaches, followed by biophysical and biochemical methods, to identify targets for Mtb phenotypic hits deposited in the ChEMBL database [11]. In this work, EthR and InhA emerged as potential targets for many of the hits, and some of them displayed activity through both targets. From the 35 predicted EthR inhibitors 25 displayed an inhibition of better than 50%, of which eight showed an IC50 better than 50 μM against Mtb EthR and three were confirmed to be also active against InhA. Further the EthR-ligand complexes were validated using X-ray crystallography in the Blundell laboratory to give new crystal structures which were deposited in the Protein Data Bank. These results provide new lead compounds that could be further developed into highly active ligands of EthR and InhA and enhance treatment of drug-resistant TB.
A comprehensive understanding of the structural proteomes of mycobacteria is essential for novel drug discovery and elucidating the roles of mutations in drug resistance. Most researchers begin by defining the 3D-structure using X-ray crystallography, NMR or increasingly cryo-EM. For phenotypic screening and understanding off-target hits, where the target is not identified, prior knowledge of the structures of all gene products in the target organism is helpful. This has stimulated the establishment of several consortia in what is usually known as structural genomics, but might more appropriately termed “structural proteomics”.
The Structural Genomics Consortium (SGC) [39] which has focused on proteins of interest to medicine, has impressive achievements, in 2011 defining ~40% of the structures of proteins from human parasites deposited in the PDB [40]. The Tuberculosis Structural Genomics Consortium (TBSGC), an international collaboration involving 53 countries, has focused on 3D structures of Mtb [40]. This activity and others working on Mtb proteomes have deposited 2274 structures in the PDB, but still representing less than 583 gene products, only 13.97% of genome. Although this is a small percentage, it compares impressively with knowledge of protein structures of two other mycobacterial pathogens where there is great clinical interest: for M. leprae causing leprosy there are experimentally-defined 3D structures for 15 gene products and for M. abscessus, a free living Mycobacterium, which is a growing challenge for cystic fibrosis patients, there are 53 experimentally-defined 3D structures in the PDB.
Comparative modelling proteins, based on the fold recognition and structural alignment with the closest homologues that have experimentally solved structures, began using interactive graphics in the 1970s [41, 42, 43]. The development of automated modelling software began in the 1980s, initially with Composer [44] and later developed with Comparer [45] and Modeller [46], based on satisfaction of 3D restraints derived from structurally aligned homologues. Modeller has now been cited ~10,500 times in the literature!
Rapid progress in this and other related software coupled with increasing computing power has enabled genome scale prediction of protein structures, as a viable alternative to experimental determination. In order to construct computational models of all gene products, which we here refer to as the structural proteome, we identify templates by a sequence-structure homology search using Fugue [47], which uses local-structural-environment-specific substitution tables to predict the likelihood of a common 3D structure. We have incorporated Fugue into a pipeline (Vivace), in which templates are selected from TOCCATA (Ochoa Montaño and Blundell, unpublished), a database of consensus profiles built from CATH 3.5 [48] and SCOP 1.75A [49] based classification of proteins structures (PDB files). PDBs within each profile are clustered based on sequence similarity using CD-HIT [50] and structures are aligned using BATON, a modified version of COMPARER [45]. After further optimization of the clusters by discarding templates with more than 20% difference in sequence identity to the maximum hit, remaining templates are classified into states based on ligand binding and oligomerization. Five different states, known as “liganded-monomeric,” “liganded-complexed,” “apo-monomeric,” “apo-complexed” and “any,” are generated in each profile hit. Models are built in each of these states using Modeller 9.10 [46] and refined. Later NDOPE, GA341 [51] Molprobity [52] and SSAG [53] are used to determine the quality of the models.
The first application of this approach was to construct the Chopin Database (
Similar models of the structural proteome for M. abscessus (Skwark et al., unpublished) and M. leprae (Vedithi et al., unpublished) have been developed in the group. In M. leprae, of the 1615 gene products, templates were identified for 1429 gene products and we were able to model 1161 proteins with high confidence. A total of 36,408 models were built in different ligand bound and oligomeric states for the 1161 proteins. The distribution of Fugue Z score across models indicates that only 4% of the proteome has no hits and 15% has poor scores. ~80% of the proteome has acceptable and good hits, and the corresponding Z scores. Around 47% of the protein queries identified templates with identity and coverage greater than 40 and 67% of the models in the proteome are of best quality as estimated by NDOPE, GA341, Molprobity and Secondary Structure Agreement (SSAG).
Current work on structural proteomes includes efforts to extend the modelling pipeline to homo-oligomeric (and eventually hetero-oligomeric) structures using comparative approaches (Malhotra et al., unpublished), extending models and improving models of small molecule complexes, and linking individual protein structures into the metabolic networks and interactions in the cell (Bannerman et al., unpublished). An example of an oligomeric structure is CTP-synthase, encoded by PyrG, which is an essential gene in Mtb identified by transposon saturation mutagenesis [54] and catalyses ATP-dependent amination of UTP to CTP with either L-glutamine or ammonia. The allosteric effector GTP functions by stabilising the protein conformation that binds to the tetrahedral intermediates formed during glutamine hydrolysis. Its closest homologue in M. leprae ML1363 is a target of choice and was modelled using Vivace during the proteome modelling exercise. We modelled the apomeric and ligand bound states of the model and oligomerized the protomer using our inhouse oligomerization pipeline. The protomeric and oligomeric states are depicted in Figure 3A and B.
(A) Protomeric model of PyrG (CTP-Synthase) of M. leprae modelled with a quality of 4.25 (best). (B) Homo-8-mer of PyrG of M. leprae modelled with a quality of 4.25 (best).
The models were built by using templates PDB-IDs: 4zdI and 4zdK for PyrG of Mtb [55]. Both the templates are 89% identical and 100% coverage to the query sequence. The superposition of the models with the templates indicated a root mean square deviation (RMSD) of 0.758.
We have also spent time over 2 decades analysing the impacts of mutations evident in the increasing wealth of available genome sequences for pathogenic mycobacteria and cancers. We originally developed SDM [56] in 1997, a method depending on statistical analysis of environment-dependent amino-acid substitution tables [57, 58]. In 2013 machine learning was introduced with the arrival of Douglas Pires in Cambridge, developing first mCSM for stability [59] followed by several “flavours” including mCSM-PPI for impacts on protein-protein interactions, mCSM-NA [60] for nucleic acid interactions and mCSM-lig for impacts on small-molecule ligand interactions useful for understanding drug resistance [61]. A critical part of using machine learning is to have an extensive database of experimentally-defined impacts of mutations on stability and interactions, such as Platinum by David Ascher when in Cambridge [62], a database of experimentally measured effects of mutations on structurally defined protein-ligand complexes that was developed for mCSM-lig. These two structural approaches to predicting the impacts of mutations (SDM & mCSM) have proved complementary and more reliable than most sequence-only methods. They also allow the application of saturation mutagenesis, facilitating in silico systematic analysis of mutations [63], an approach now being adopted to whole proteomes where every residue in each of the proteins in the proteome is mutated to all the other 19 amino acids and the effects of the mutations are measured using various methods mentioned above. In structure-guided fragment-based drug discovery, this provides comprehensive information on the regions of the protein that are less likely to lead to drug resistance and therefore can be probed by elaboration of fragments/small molecules. We performed saturation mutagenesis on the drug targets in M. leprae for leprosy and the average or highest impact a mutation can induce in each residue position is depicted on the structure (Figure 4).
Indicates the maximum destabilising effect a mutation can induce on the stability of RNA-polymerase β-subunit of M. leprae (target for rifampin) measured by mCSM-stability.
Although comparative modelling of homologues in complex with ligands can often give clues about active sites, cofactor binding and substrate or other ligand binding sites, this is not always possible. In order to indicate putative binding sites in the absence of appropriate experimental data, we have exploited cavity-defining software such as VolSite [64] for novel binding site description together with an alignment and comparison tool (Shaper) [65]. We have used FuzCav, a novel alignment-free high-throughput algorithm to compute pairwise similarities between protein-ligand binding sites [66] and GHECOM [67], to study the small pockets that often characterise protein-protein and protein-peptide interactions.
Further to the identification of cavities and pockets, it is also useful to be able to identify hotspots, region(s) of the binding site defined as a major contributor to the binding free energy, and often characterised by their ability to bind fragment-sized organic molecules in well-defined orientations. The usual understanding is that the fragment, with a mixed polar and hydrophobic character, can displace an “unhappy water.” We have tried to mimic this in silico by using SuperStar [68] to generate atomic interaction propensities on a grid. We then carry out a search with three fragments, each having a six-membered carbon ring, but having a donor, acceptor or a non-polar substituent. The resulting map is convoluted with an estimate of the depth below the surface, which generally appears to correlate with favourable entropic gain on water release on binding of a ligand [69]. The hotspot maps, computed in this way and indicating donor, acceptor and lipophilic interactions correlate well with experimental binding sites of fragments that can be elaborated in fragment-based discovery. For the ligand bound structures, lower contouring can provide “warm spots” for the binding sites, indicating possibilities for elaborating the fragment in the binding pocket.
The models of individual molecules of the modelled proteome can be individually decorated with the hotspot maps. They give a good indication of the known functional sites on experimentally defined structures of proteins, often demonstrating that a functional site comprises several hotspots involved in binding substrates and cofactors. They also provide a good indication of the location of allosteric sites [70].
In summary we can move from the study of individual targets to an understanding of the majority of targets coded by the genome. Indeed, we can build 3D structures for a majority of the genes, so providing a model of the “structural proteome”. Hotspots and cavities provide a basis for identification of the ligandability of putative binding sites and have been used in our group to predict pharmacophores that can be used in docking and virtual screening and so deorphaning of mycobacterial proteins.
To identify druggable proteins from the structural proteome, we have adopted a hierarchal selection process wherein chokepoint analysis is initially performed to identify metabolic reactions that are critical to cell survival. Gene products identified in this screen are later subjected to essentiality analysis using either flux balance analysis (FBA) based models or by data from the transposon saturation mutagenesis experiments in the literature. Genes that are essential are chosen at this stage and understanding of the gene expression profiles in different growth conditions is analysed. Genes whose expression is condition specific are excluded. Later for the selected genes, the structural information of the corresponding proteins is analysed in the context of prior knowledge and attempts in drug discovery, druggable pockets and fragment hotspots maps, small molecule bound states, non-human homologue, non-homologous to human microbiome, cellular localization and biochemical properties of the proteins. Structure-guided virtual screening is performed on the selected drug targets with a choice of fragment and compound libraries using CCDC Gold (The Cambridge Crystallographic Data Centre) [71]. Best poses with good scores lead the experimental process of structure-guided fragment-based drug discovery.
The challenge now is to test the computational methods outlined here for identifying ligands and understanding the druggability of the proteome—several thousand gene products from the whole genome of Mtb. We can then begin to assess the degree to which we can de-orphan the many Mtb proteins that have until now not featured as targets in the worldwide efforts to combat the global challenge of TB to the health and well-being of human kind.
LYB and GCM are grateful to Chinhoyi University of Technology for their support in introducing computational drug discovery and development research work at the University and all our collaborators. TLB and SCV thank the Gates Foundation, the Cystic Fibrosis Trust and the American Leprosy Mission for their funding of computational and experimental work on approaches to combating disease from mycobacterial infections. They also thank colleagues in Cambridge and elsewhere who have contributed over the years to our efforts to develop new approaches to structural biology, computational bioinformatics and drug discovery.
"I work with IntechOpen for a number of reasons: their professionalism, their mission in support of Open Access publishing, and the quality of their peer-reviewed publications, but also because they believe in equality. Throughout the world, we are seeing progress in attracting, retaining, and promoting women in STEMM. IntechOpen are certainly supporting this work globally by empowering all scientists and ensuring that women are encouraged and enabled to publish and take leading roles within the scientific community." Dr. Catrin Rutland, University of Nottingham, UK
",metaTitle:"Advantages of Publishing with IntechOpen",metaDescription:"We have more than a decade of experience in Open Access publishing. \n\n ",metaKeywords:null,canonicalURL:null,contentRaw:'[{"type":"htmlEditorComponent","content":"We have more than a decade of experience in Open Access publishing. The advantages of publishing with IntechOpen include:
\\n\\nOur platform – IntechOpen is the world’s leading publisher of OA books, built by scientists, for scientists.
\\n\\nOur reputation – Everything we publish goes through a two-stage peer review process. We’re proud to count Nobel laureates among our esteemed authors. We meet European Commission standards for funding, and the research we’ve published has been funded by the Bill and Melinda Gates Foundation and the Wellcome Trust, among others. IntechOpen is a member of all relevant trade associations (including the STM Association and the Association of Learned and Professional Society Publishers) and has a selection of books indexed in Web of Science's Book Citation Index.
\\n\\nOur expertise – We’ve published more than 4,500 books by more than 118,000 authors and editors.
\\n\\nOur reach – Our books have more than 130 million downloads and more than 146,150 Web of Science citations. We increase citations via indexing in all the major databases, including the Book Citation Index at Web of Science and Google Scholar.
\\n\\nOur services – The support we offer our authors and editors is second to none. Each book in our program receives the following:
\\n\\nOur end-to-end publishing service frees our authors and editors to focus on what matters: research. We empower them to shape their fields and connect with the global scientific community.
\\n\\n"In developing countries until now, advancement in science has been very limited, because insufficient economic resources are dedicated to science and education. These limitations are more marked when the scientists are women. In order to develop science in the poorest countries and decrease the gender gap that exists in scientific fields, Open Access networks like IntechOpen are essential. Free access to scientific research could contribute to ameliorating difficult life conditions and breaking down barriers." Marquidia Pacheco, National Institute for Nuclear Research (ININ), Mexico
\\n\\nInterested? Contact Ana Pantar (book.idea@intechopen.com) for more information.
\\n"}]'},components:[{type:"htmlEditorComponent",content:'We have more than a decade of experience in Open Access publishing. The advantages of publishing with IntechOpen include:
\n\nOur platform – IntechOpen is the world’s leading publisher of OA books, built by scientists, for scientists.
\n\nOur reputation – Everything we publish goes through a two-stage peer review process. We’re proud to count Nobel laureates among our esteemed authors. We meet European Commission standards for funding, and the research we’ve published has been funded by the Bill and Melinda Gates Foundation and the Wellcome Trust, among others. IntechOpen is a member of all relevant trade associations (including the STM Association and the Association of Learned and Professional Society Publishers) and has a selection of books indexed in Web of Science's Book Citation Index.
\n\nOur expertise – We’ve published more than 4,500 books by more than 118,000 authors and editors.
\n\nOur reach – Our books have more than 130 million downloads and more than 146,150 Web of Science citations. We increase citations via indexing in all the major databases, including the Book Citation Index at Web of Science and Google Scholar.
\n\nOur services – The support we offer our authors and editors is second to none. Each book in our program receives the following:
\n\nOur end-to-end publishing service frees our authors and editors to focus on what matters: research. We empower them to shape their fields and connect with the global scientific community.
\n\n"In developing countries until now, advancement in science has been very limited, because insufficient economic resources are dedicated to science and education. These limitations are more marked when the scientists are women. In order to develop science in the poorest countries and decrease the gender gap that exists in scientific fields, Open Access networks like IntechOpen are essential. Free access to scientific research could contribute to ameliorating difficult life conditions and breaking down barriers." Marquidia Pacheco, National Institute for Nuclear Research (ININ), Mexico
\n\nInterested? Contact Ana Pantar (book.idea@intechopen.com) for more information.
\n'}]},successStories:{items:[]},authorsAndEditors:{filterParams:{sort:"featured,name"},profiles:[{id:"58592",title:"Dr.",name:"Arun",middleName:null,surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/58592/images/1664_n.jpg",biography:"Arun K. Shanker is serving as a Principal Scientist (Plant Physiology) with the Indian Council of Agricultural Research (ICAR) at the Central Research Institute for Dryland Agriculture in Hyderabad, India. He is working with the ICAR as a full time researcher since 1993 and has since earned his Advanced degree in Crop Physiology while in service. He has been awarded the prestigious Member of the Royal Society of Chemistry (MRSC), by the Royal Society of Chemistry, London in 2015. Presently he is working on systems biology approach to study the mechanism of abiotic stress tolerance in crops. His main focus now is to unravel the mechanism of drought and heat stress response in plants to tackle climate change related threats in agriculture.",institutionString:null,institution:{name:"Indian Council of Agricultural Research",country:{name:"India"}}},{id:"4782",title:"Prof.",name:"Bishnu",middleName:"P",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/4782/images/system/4782.jpg",biography:"Bishnu P. Pal is Professor of Physics at Mahindra École\nCentrale Hyderabad India since July 1st 2014 after retirement\nas Professor of Physics from IIT Delhi; Ph.D.’1975 from IIT\nDelhi; Fellow of OSA and SPIE; Senior Member IEEE;\nHonorary Foreign Member Royal Norwegian Society for\nScience and Arts; Member OSA Board of Directors (2009-\n11); Distinguished Lecturer IEEE Photonics Society (2005-\n07).",institutionString:null,institution:{name:"Indian Institute of Technology Delhi",country:{name:"India"}}},{id:"69653",title:"Dr.",name:"Chusak",middleName:null,surname:"Limsakul",slug:"chusak-limsakul",fullName:"Chusak Limsakul",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Prince of Songkla University",country:{name:"Thailand"}}},{id:"75563",title:"Dr.",name:"Farzana Khan",middleName:null,surname:"Perveen",slug:"farzana-khan-perveen",fullName:"Farzana Khan Perveen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/75563/images/system/75563.png",biography:"Dr Farzana Khan Perveen (FLS; Gold-Medallist) obtained her BSc (Hons) and MSc (Zoology: Entomology) from the University of Karachi, MAS (Monbush-Scholar; Agriculture: Agronomy) and from the Nagoya University, Japan, and PhD (Research and Course-works from the Nagoya University; Toxicology) degree from the University of Karachi. She is Founder/Chairperson of the Department of Zoology (DOZ) and Ex-Controller of Examinations at Shaheed Benazir Bhutto University (SBBU) and Ex-Founder/ Ex-Chairperson of DOZ, Hazara University and Kohat University of Science & Technology. \nShe is the author of 150 high impact research papers, 135 abstracts, 4 authored books and 8 chapters. She is the editor of 5 books and she supervised BS(4), MSc(50), MPhil(40), and Ph.D. (1) students. She has organized and participated in numerous international and national conferences and received multiple awards and fellowships. She is a member of research societies, editorial boards of Journals, and World-Commission on Protected Areas, International Union for Conservation of Nature. Her fields of interest are Entomology, Toxicology, Forensic Entomology, and Zoology.",institutionString:"Shaheed Benazir Bhutto University",institution:{name:"Shaheed Benazir Bhutto University",country:{name:"Pakistan"}}},{id:"23804",title:"Dr.",name:"Hamzah",middleName:null,surname:"Arof",slug:"hamzah-arof",fullName:"Hamzah Arof",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/23804/images/5492_n.jpg",biography:"Hamzah Arof received his BSc from Michigan State University, and PhD from the University of Wales. Both degrees were in electrical engineering. His current research interests include signal processing and photonics. Currently he is affiliated with the Department of Electrical Engineering, University of Malaya, Malaysia.",institutionString:null,institution:{name:"University of Malaya",country:{name:"Malaysia"}}},{id:"41989",title:"Prof.",name:"He",middleName:null,surname:"Tian",slug:"he-tian",fullName:"He Tian",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"East China University of Science and Technology",country:{name:"China"}}},{id:"33351",title:null,name:"Hendra",middleName:null,surname:"Hermawan",slug:"hendra-hermawan",fullName:"Hendra Hermawan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/33351/images/168_n.jpg",biography:null,institutionString:null,institution:{name:"Institut Teknologi Bandung",country:{name:"Indonesia"}}},{id:"11981",title:"Prof.",name:"Hiroshi",middleName:null,surname:"Ishiguro",slug:"hiroshi-ishiguro",fullName:"Hiroshi Ishiguro",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Osaka University",country:{name:"Japan"}}},{id:"45747",title:"Dr.",name:"Hsin-I",middleName:null,surname:"Chang",slug:"hsin-i-chang",fullName:"Hsin-I Chang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/no_image.jpg",biography:null,institutionString:null,institution:{name:"National Chiayi University",country:{name:"Taiwan"}}},{id:"61581",title:"Dr.",name:"Joy Rizki Pangestu",middleName:null,surname:"Djuansjah",slug:"joy-rizki-pangestu-djuansjah",fullName:"Joy Rizki Pangestu Djuansjah",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/61581/images/237_n.jpg",biography:null,institutionString:null,institution:{name:"University of Technology Malaysia",country:{name:"Malaysia"}}},{id:"94249",title:"Prof.",name:"Junji",middleName:null,surname:"Kido",slug:"junji-kido",fullName:"Junji Kido",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Yamagata University",country:{name:"Japan"}}},{id:"12009",title:"Dr.",name:"Ki Young",middleName:null,surname:"Kim",slug:"ki-young-kim",fullName:"Ki Young Kim",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/12009/images/system/12009.jpg",biography:"Http://m80.knu.ac.kr/~doors",institutionString:null,institution:{name:"National Cheng Kung University",country:{name:"Taiwan"}}}],filtersByRegion:[{group:"region",caption:"North America",value:1,count:5763},{group:"region",caption:"Middle and South America",value:2,count:5227},{group:"region",caption:"Africa",value:3,count:1717},{group:"region",caption:"Asia",value:4,count:10365},{group:"region",caption:"Australia and Oceania",value:5,count:897},{group:"region",caption:"Europe",value:6,count:15784}],offset:12,limit:12,total:10365},chapterEmbeded:{data:{}},editorApplication:{success:null,errors:{}},ofsBooks:{filterParams:{topicId:"6"},books:[{type:"book",id:"8977",title:"Protein Kinase - New Opportunities, Challenges and Future Perspectives",subtitle:null,isOpenForSubmission:!0,hash:"6d200cc031706a565b554fdb1c478901",slug:null,bookSignature:"Dr. Rajesh Kumar Singh",coverURL:"https://cdn.intechopen.com/books/images_new/8977.jpg",editedByType:null,editors:[{id:"329385",title:"Dr.",name:"Rajesh",surname:"Singh",slug:"rajesh-singh",fullName:"Rajesh Singh"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9659",title:"Fibroblasts - Advances in Cancer, Autoimmunity and Inflammation",subtitle:null,isOpenForSubmission:!0,hash:"926fa6446f6befbd363fc74971a56de2",slug:null,bookSignature:"Ph.D. Mojca Frank Bertoncelj and Ms. Katja Lakota",coverURL:"https://cdn.intechopen.com/books/images_new/9659.jpg",editedByType:null,editors:[{id:"328755",title:"Ph.D.",name:"Mojca",surname:"Frank Bertoncelj",slug:"mojca-frank-bertoncelj",fullName:"Mojca Frank Bertoncelj"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10797",title:"Cell Culture",subtitle:null,isOpenForSubmission:!0,hash:"2c628f4757f9639a4450728d839a7842",slug:null,bookSignature:"Prof. Xianquan Zhan",coverURL:"https://cdn.intechopen.com/books/images_new/10797.jpg",editedByType:null,editors:[{id:"223233",title:"Prof.",name:"Xianquan",surname:"Zhan",slug:"xianquan-zhan",fullName:"Xianquan Zhan"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10800",title:"Ligase",subtitle:null,isOpenForSubmission:!0,hash:"1f10ff112edb1fec24379dac85ef3b5b",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/10800.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10801",title:"Uric Acid",subtitle:null,isOpenForSubmission:!0,hash:"d947ab87019e69ab11aa597edbacc018",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/10801.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10837",title:"Peroxisomes",subtitle:null,isOpenForSubmission:!0,hash:"0014b09d4b35bb4d7f52ca0b3641cda1",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/10837.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10838",title:"Ion Channels",subtitle:null,isOpenForSubmission:!0,hash:"048017b227b3bdfd0d33a49bac63c915",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/10838.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10840",title:"Benzimidazole",subtitle:null,isOpenForSubmission:!0,hash:"9fe810233f92a9c454c624aec634316f",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/10840.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10841",title:"Hydrolases",subtitle:null,isOpenForSubmission:!0,hash:"64617cf21bf1e47170bb2bcf31b1fc37",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/10841.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],filtersByTopic:[{group:"topic",caption:"Agricultural and Biological Sciences",value:5,count:13},{group:"topic",caption:"Biochemistry, Genetics and Molecular Biology",value:6,count:3},{group:"topic",caption:"Business, Management and Economics",value:7,count:1},{group:"topic",caption:"Chemistry",value:8,count:6},{group:"topic",caption:"Computer and Information Science",value:9,count:6},{group:"topic",caption:"Earth and Planetary Sciences",value:10,count:7},{group:"topic",caption:"Engineering",value:11,count:14},{group:"topic",caption:"Environmental Sciences",value:12,count:2},{group:"topic",caption:"Immunology and Microbiology",value:13,count:3},{group:"topic",caption:"Materials Science",value:14,count:4},{group:"topic",caption:"Mathematics",value:15,count:1},{group:"topic",caption:"Medicine",value:16,count:27},{group:"topic",caption:"Neuroscience",value:18,count:1},{group:"topic",caption:"Pharmacology, Toxicology and Pharmaceutical Science",value:19,count:2},{group:"topic",caption:"Physics",value:20,count:2},{group:"topic",caption:"Psychology",value:21,count:4},{group:"topic",caption:"Social Sciences",value:23,count:2},{group:"topic",caption:"Technology",value:24,count:1},{group:"topic",caption:"Veterinary Medicine and Science",value:25,count:1}],offset:12,limit:12,total:9},popularBooks:{featuredBooks:[{type:"book",id:"9385",title:"Renewable Energy",subtitle:"Technologies and Applications",isOpenForSubmission:!1,hash:"a6b446d19166f17f313008e6c056f3d8",slug:"renewable-energy-technologies-and-applications",bookSignature:"Tolga Taner, Archana Tiwari and Taha Selim Ustun",coverURL:"https://cdn.intechopen.com/books/images_new/9385.jpg",editors:[{id:"197240",title:"Associate Prof.",name:"Tolga",middleName:null,surname:"Taner",slug:"tolga-taner",fullName:"Tolga Taner"}],equalEditorOne:{id:"186791",title:"Dr.",name:"Archana",middleName:null,surname:"Tiwari",slug:"archana-tiwari",fullName:"Archana Tiwari",profilePictureURL:"https://mts.intechopen.com/storage/users/186791/images/system/186791.png",biography:"Dr. Archana Tiwari is Associate Professor at Amity University, India. Her research interests include renewable sources of energy from microalgae and further utilizing the residual biomass for the generation of value-added products, bioremediation through microalgae and microbial consortium, antioxidative enzymes and stress, and nutraceuticals from microalgae. She has been working on algal biotechnology for the last two decades. She has published her research in many international journals and has authored many books and chapters with renowned publishing houses. She has also delivered talks as an invited speaker at many national and international conferences. Dr. Tiwari is the recipient of several awards including Researcher of the Year and Distinguished Scientist.",institutionString:"Amity University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"Amity University",institutionURL:null,country:{name:"India"}}},equalEditorTwo:{id:"197609",title:"Prof.",name:"Taha Selim",middleName:null,surname:"Ustun",slug:"taha-selim-ustun",fullName:"Taha Selim Ustun",profilePictureURL:"https://mts.intechopen.com/storage/users/197609/images/system/197609.jpeg",biography:"Dr. Taha Selim Ustun received a Ph.D. in Electrical Engineering from Victoria University, Melbourne, Australia. He is a researcher with the Fukushima Renewable Energy Institute, AIST (FREA), where he leads the Smart Grid Cybersecurity Laboratory. Prior to that, he was a faculty member with the School of Electrical and Computer Engineering, Carnegie Mellon University, Pittsburgh, PA, USA. His current research interests include power systems protection, communication in power networks, distributed generation, microgrids, electric vehicle integration, and cybersecurity in smart grids. He serves on the editorial boards of IEEE Access, IEEE Transactions on Industrial Informatics, Energies, Electronics, Electricity, World Electric Vehicle and Information journals. Dr. Ustun is a member of the IEEE 2004 and 2800, IEC Renewable Energy Management WG 8, and IEC TC 57 WG17. He has been invited to run specialist courses in Africa, India, and China. He has delivered talks for the Qatar Foundation, the World Energy Council, the Waterloo Global Science Initiative, and the European Union Energy Initiative (EUEI). His research has attracted funding from prestigious programs in Japan, Australia, the European Union, and North America.",institutionString:"Fukushima Renewable Energy Institute, AIST (FREA)",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"National Institute of Advanced Industrial Science and Technology",institutionURL:null,country:{name:"Japan"}}},equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8985",title:"Natural Resources Management and Biological Sciences",subtitle:null,isOpenForSubmission:!1,hash:"5c2e219a6c021a40b5a20c041dea88c4",slug:"natural-resources-management-and-biological-sciences",bookSignature:"Edward R. Rhodes and Humood Naser",coverURL:"https://cdn.intechopen.com/books/images_new/8985.jpg",editors:[{id:"280886",title:"Prof.",name:"Edward R",middleName:null,surname:"Rhodes",slug:"edward-r-rhodes",fullName:"Edward R Rhodes"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9027",title:"Human Blood Group Systems and Haemoglobinopathies",subtitle:null,isOpenForSubmission:!1,hash:"d00d8e40b11cfb2547d1122866531c7e",slug:"human-blood-group-systems-and-haemoglobinopathies",bookSignature:"Osaro Erhabor and Anjana Munshi",coverURL:"https://cdn.intechopen.com/books/images_new/9027.jpg",editors:[{id:"35140",title:null,name:"Osaro",middleName:null,surname:"Erhabor",slug:"osaro-erhabor",fullName:"Osaro Erhabor"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7841",title:"New Insights Into Metabolic Syndrome",subtitle:null,isOpenForSubmission:!1,hash:"ef5accfac9772b9e2c9eff884f085510",slug:"new-insights-into-metabolic-syndrome",bookSignature:"Akikazu Takada",coverURL:"https://cdn.intechopen.com/books/images_new/7841.jpg",editors:[{id:"248459",title:"Dr.",name:"Akikazu",middleName:null,surname:"Takada",slug:"akikazu-takada",fullName:"Akikazu Takada"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8620",title:"Mining Techniques",subtitle:"Past, Present and Future",isOpenForSubmission:!1,hash:"b65658f81d14e9e57e49377869d3a575",slug:"mining-techniques-past-present-and-future",bookSignature:"Abhay Soni",coverURL:"https://cdn.intechopen.com/books/images_new/8620.jpg",editors:[{id:"271093",title:"Dr.",name:"Abhay",middleName:null,surname:"Soni",slug:"abhay-soni",fullName:"Abhay Soni"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8558",title:"Aerodynamics",subtitle:null,isOpenForSubmission:!1,hash:"db7263fc198dfb539073ba0260a7f1aa",slug:"aerodynamics",bookSignature:"Mofid Gorji-Bandpy and Aly-Mousaad Aly",coverURL:"https://cdn.intechopen.com/books/images_new/8558.jpg",editors:[{id:"35542",title:"Prof.",name:"Mofid",middleName:null,surname:"Gorji-Bandpy",slug:"mofid-gorji-bandpy",fullName:"Mofid Gorji-Bandpy"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9668",title:"Chemistry and Biochemistry of Winemaking, Wine Stabilization and Aging",subtitle:null,isOpenForSubmission:!1,hash:"c5484276a314628acf21ec1bdc3a86b9",slug:"chemistry-and-biochemistry-of-winemaking-wine-stabilization-and-aging",bookSignature:"Fernanda Cosme, Fernando M. Nunes and Luís Filipe-Ribeiro",coverURL:"https://cdn.intechopen.com/books/images_new/9668.jpg",editors:[{id:"186819",title:"Prof.",name:"Fernanda",middleName:null,surname:"Cosme",slug:"fernanda-cosme",fullName:"Fernanda Cosme"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7847",title:"Medical Toxicology",subtitle:null,isOpenForSubmission:!1,hash:"db9b65bea093de17a0855a1b27046247",slug:"medical-toxicology",bookSignature:"Pınar Erkekoglu and Tomohisa Ogawa",coverURL:"https://cdn.intechopen.com/books/images_new/7847.jpg",editors:[{id:"109978",title:"Prof.",name:"Pınar",middleName:null,surname:"Erkekoglu",slug:"pinar-erkekoglu",fullName:"Pınar Erkekoglu"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9660",title:"Inland Waters",subtitle:"Dynamics and Ecology",isOpenForSubmission:!1,hash:"975c26819ceb11a926793bc2adc62bd6",slug:"inland-waters-dynamics-and-ecology",bookSignature:"Adam Devlin, Jiayi Pan and Mohammad Manjur Shah",coverURL:"https://cdn.intechopen.com/books/images_new/9660.jpg",editors:[{id:"280757",title:"Dr.",name:"Adam",middleName:"Thomas",surname:"Devlin",slug:"adam-devlin",fullName:"Adam Devlin"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9731",title:"Oxidoreductase",subtitle:null,isOpenForSubmission:!1,hash:"852e6f862c85fc3adecdbaf822e64e6e",slug:"oxidoreductase",bookSignature:"Mahmoud Ahmed Mansour",coverURL:"https://cdn.intechopen.com/books/images_new/9731.jpg",editors:[{id:"224662",title:"Prof.",name:"Mahmoud Ahmed",middleName:null,surname:"Mansour",slug:"mahmoud-ahmed-mansour",fullName:"Mahmoud Ahmed Mansour"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9864",title:"Hydrology",subtitle:null,isOpenForSubmission:!1,hash:"02925c63436d12e839008c793a253310",slug:"hydrology",bookSignature:"Theodore V. Hromadka II and Prasada Rao",coverURL:"https://cdn.intechopen.com/books/images_new/9864.jpg",editors:[{id:"181008",title:"Dr.",name:"Theodore V.",middleName:"V.",surname:"Hromadka II",slug:"theodore-v.-hromadka-ii",fullName:"Theodore V. Hromadka II"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9083",title:"Rodents",subtitle:null,isOpenForSubmission:!1,hash:"480148de5ecf236b3e0860fc3954b2d4",slug:"rodents",bookSignature:"Loth S. Mulungu",coverURL:"https://cdn.intechopen.com/books/images_new/9083.jpg",editors:[{id:"108433",title:"Dr.",name:"Loth S.",middleName:null,surname:"Mulungu",slug:"loth-s.-mulungu",fullName:"Loth S. Mulungu"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}}],offset:12,limit:12,total:5220},hotBookTopics:{hotBooks:[],offset:0,limit:12,total:null},publish:{},publishingProposal:{success:null,errors:{}},books:{featuredBooks:[{type:"book",id:"9385",title:"Renewable Energy",subtitle:"Technologies and Applications",isOpenForSubmission:!1,hash:"a6b446d19166f17f313008e6c056f3d8",slug:"renewable-energy-technologies-and-applications",bookSignature:"Tolga Taner, Archana Tiwari and Taha Selim Ustun",coverURL:"https://cdn.intechopen.com/books/images_new/9385.jpg",editors:[{id:"197240",title:"Associate Prof.",name:"Tolga",middleName:null,surname:"Taner",slug:"tolga-taner",fullName:"Tolga Taner"}],equalEditorOne:{id:"186791",title:"Dr.",name:"Archana",middleName:null,surname:"Tiwari",slug:"archana-tiwari",fullName:"Archana Tiwari",profilePictureURL:"https://mts.intechopen.com/storage/users/186791/images/system/186791.jpg",biography:"Dr. Archana Tiwari is Associate Professor at Amity University, India. Her research interests include renewable sources of energy from microalgae and further utilizing the residual biomass for the generation of value-added products, bioremediation through microalgae and microbial consortium, antioxidative enzymes and stress, and nutraceuticals from microalgae. She has been working on algal biotechnology for the last two decades. She has published her research in many international journals and has authored many books and chapters with renowned publishing houses. She has also delivered talks as an invited speaker at many national and international conferences. Dr. Tiwari is the recipient of several awards including Researcher of the Year and Distinguished Scientist.",institutionString:"Amity University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"Amity University",institutionURL:null,country:{name:"India"}}},equalEditorTwo:{id:"197609",title:"Prof.",name:"Taha Selim",middleName:null,surname:"Ustun",slug:"taha-selim-ustun",fullName:"Taha Selim Ustun",profilePictureURL:"https://mts.intechopen.com/storage/users/197609/images/system/197609.jpeg",biography:"Dr. Taha Selim Ustun received a Ph.D. in Electrical Engineering from Victoria University, Melbourne, Australia. He is a researcher with the Fukushima Renewable Energy Institute, AIST (FREA), where he leads the Smart Grid Cybersecurity Laboratory. Prior to that, he was a faculty member with the School of Electrical and Computer Engineering, Carnegie Mellon University, Pittsburgh, PA, USA. His current research interests include power systems protection, communication in power networks, distributed generation, microgrids, electric vehicle integration, and cybersecurity in smart grids. He serves on the editorial boards of IEEE Access, IEEE Transactions on Industrial Informatics, Energies, Electronics, Electricity, World Electric Vehicle and Information journals. Dr. Ustun is a member of the IEEE 2004 and 2800, IEC Renewable Energy Management WG 8, and IEC TC 57 WG17. He has been invited to run specialist courses in Africa, India, and China. He has delivered talks for the Qatar Foundation, the World Energy Council, the Waterloo Global Science Initiative, and the European Union Energy Initiative (EUEI). His research has attracted funding from prestigious programs in Japan, Australia, the European Union, and North America.",institutionString:"Fukushima Renewable Energy Institute, AIST (FREA)",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"National Institute of Advanced Industrial Science and Technology",institutionURL:null,country:{name:"Japan"}}},equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8985",title:"Natural Resources Management and Biological Sciences",subtitle:null,isOpenForSubmission:!1,hash:"5c2e219a6c021a40b5a20c041dea88c4",slug:"natural-resources-management-and-biological-sciences",bookSignature:"Edward R. Rhodes and Humood Naser",coverURL:"https://cdn.intechopen.com/books/images_new/8985.jpg",editors:[{id:"280886",title:"Prof.",name:"Edward R",middleName:null,surname:"Rhodes",slug:"edward-r-rhodes",fullName:"Edward R Rhodes"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9027",title:"Human Blood Group Systems and Haemoglobinopathies",subtitle:null,isOpenForSubmission:!1,hash:"d00d8e40b11cfb2547d1122866531c7e",slug:"human-blood-group-systems-and-haemoglobinopathies",bookSignature:"Osaro Erhabor and Anjana Munshi",coverURL:"https://cdn.intechopen.com/books/images_new/9027.jpg",editors:[{id:"35140",title:null,name:"Osaro",middleName:null,surname:"Erhabor",slug:"osaro-erhabor",fullName:"Osaro Erhabor"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7841",title:"New Insights Into Metabolic Syndrome",subtitle:null,isOpenForSubmission:!1,hash:"ef5accfac9772b9e2c9eff884f085510",slug:"new-insights-into-metabolic-syndrome",bookSignature:"Akikazu Takada",coverURL:"https://cdn.intechopen.com/books/images_new/7841.jpg",editors:[{id:"248459",title:"Dr.",name:"Akikazu",middleName:null,surname:"Takada",slug:"akikazu-takada",fullName:"Akikazu Takada"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8558",title:"Aerodynamics",subtitle:null,isOpenForSubmission:!1,hash:"db7263fc198dfb539073ba0260a7f1aa",slug:"aerodynamics",bookSignature:"Mofid Gorji-Bandpy and Aly-Mousaad Aly",coverURL:"https://cdn.intechopen.com/books/images_new/8558.jpg",editors:[{id:"35542",title:"Prof.",name:"Mofid",middleName:null,surname:"Gorji-Bandpy",slug:"mofid-gorji-bandpy",fullName:"Mofid Gorji-Bandpy"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9668",title:"Chemistry and Biochemistry of Winemaking, Wine Stabilization and Aging",subtitle:null,isOpenForSubmission:!1,hash:"c5484276a314628acf21ec1bdc3a86b9",slug:"chemistry-and-biochemistry-of-winemaking-wine-stabilization-and-aging",bookSignature:"Fernanda Cosme, Fernando M. Nunes and Luís Filipe-Ribeiro",coverURL:"https://cdn.intechopen.com/books/images_new/9668.jpg",editors:[{id:"186819",title:"Prof.",name:"Fernanda",middleName:null,surname:"Cosme",slug:"fernanda-cosme",fullName:"Fernanda Cosme"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7847",title:"Medical Toxicology",subtitle:null,isOpenForSubmission:!1,hash:"db9b65bea093de17a0855a1b27046247",slug:"medical-toxicology",bookSignature:"Pınar Erkekoglu and Tomohisa Ogawa",coverURL:"https://cdn.intechopen.com/books/images_new/7847.jpg",editors:[{id:"109978",title:"Prof.",name:"Pınar",middleName:null,surname:"Erkekoglu",slug:"pinar-erkekoglu",fullName:"Pınar Erkekoglu"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8620",title:"Mining Techniques",subtitle:"Past, Present and Future",isOpenForSubmission:!1,hash:"b65658f81d14e9e57e49377869d3a575",slug:"mining-techniques-past-present-and-future",bookSignature:"Abhay Soni",coverURL:"https://cdn.intechopen.com/books/images_new/8620.jpg",editors:[{id:"271093",title:"Dr.",name:"Abhay",middleName:null,surname:"Soni",slug:"abhay-soni",fullName:"Abhay Soni"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9660",title:"Inland Waters",subtitle:"Dynamics and Ecology",isOpenForSubmission:!1,hash:"975c26819ceb11a926793bc2adc62bd6",slug:"inland-waters-dynamics-and-ecology",bookSignature:"Adam Devlin, Jiayi Pan and Mohammad Manjur Shah",coverURL:"https://cdn.intechopen.com/books/images_new/9660.jpg",editors:[{id:"280757",title:"Dr.",name:"Adam",middleName:"Thomas",surname:"Devlin",slug:"adam-devlin",fullName:"Adam Devlin"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9122",title:"Cosmetic Surgery",subtitle:null,isOpenForSubmission:!1,hash:"207026ca4a4125e17038e770d00ee152",slug:"cosmetic-surgery",bookSignature:"Yueh-Bih Tang",coverURL:"https://cdn.intechopen.com/books/images_new/9122.jpg",editors:[{id:"202122",title:"Prof.",name:"Yueh-Bih",middleName:null,surname:"Tang",slug:"yueh-bih-tang",fullName:"Yueh-Bih Tang"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}}],latestBooks:[{type:"book",id:"9550",title:"Entrepreneurship",subtitle:"Contemporary Issues",isOpenForSubmission:!1,hash:"9b4ac1ee5b743abf6f88495452b1e5e7",slug:"entrepreneurship-contemporary-issues",bookSignature:"Mladen Turuk",coverURL:"https://cdn.intechopen.com/books/images_new/9550.jpg",editedByType:"Edited by",editors:[{id:"319755",title:"Prof.",name:"Mladen",middleName:null,surname:"Turuk",slug:"mladen-turuk",fullName:"Mladen Turuk"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10065",title:"Wavelet Theory",subtitle:null,isOpenForSubmission:!1,hash:"d8868e332169597ba2182d9b004d60de",slug:"wavelet-theory",bookSignature:"Somayeh Mohammady",coverURL:"https://cdn.intechopen.com/books/images_new/10065.jpg",editedByType:"Edited by",editors:[{id:"109280",title:"Dr.",name:"Somayeh",middleName:null,surname:"Mohammady",slug:"somayeh-mohammady",fullName:"Somayeh Mohammady"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9313",title:"Clay Science and Technology",subtitle:null,isOpenForSubmission:!1,hash:"6fa7e70396ff10620e032bb6cfa6fb72",slug:"clay-science-and-technology",bookSignature:"Gustavo Morari Do Nascimento",coverURL:"https://cdn.intechopen.com/books/images_new/9313.jpg",editedByType:"Edited by",editors:[{id:"7153",title:"Prof.",name:"Gustavo",middleName:null,surname:"Morari Do Nascimento",slug:"gustavo-morari-do-nascimento",fullName:"Gustavo Morari Do Nascimento"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9888",title:"Nuclear Power Plants",subtitle:"The Processes from the Cradle to the Grave",isOpenForSubmission:!1,hash:"c2c8773e586f62155ab8221ebb72a849",slug:"nuclear-power-plants-the-processes-from-the-cradle-to-the-grave",bookSignature:"Nasser Awwad",coverURL:"https://cdn.intechopen.com/books/images_new/9888.jpg",editedByType:"Edited by",editors:[{id:"145209",title:"Prof.",name:"Nasser",middleName:"S",surname:"Awwad",slug:"nasser-awwad",fullName:"Nasser Awwad"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8098",title:"Resources of Water",subtitle:null,isOpenForSubmission:!1,hash:"d251652996624d932ef7b8ed62cf7cfc",slug:"resources-of-water",bookSignature:"Prathna Thanjavur Chandrasekaran, Muhammad Salik Javaid, Aftab Sadiq",coverURL:"https://cdn.intechopen.com/books/images_new/8098.jpg",editedByType:"Edited by",editors:[{id:"167917",title:"Dr.",name:"Prathna",middleName:null,surname:"Thanjavur Chandrasekaran",slug:"prathna-thanjavur-chandrasekaran",fullName:"Prathna Thanjavur Chandrasekaran"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9644",title:"Glaciers and the Polar Environment",subtitle:null,isOpenForSubmission:!1,hash:"e8cfdc161794e3753ced54e6ff30873b",slug:"glaciers-and-the-polar-environment",bookSignature:"Masaki Kanao, Danilo Godone and Niccolò Dematteis",coverURL:"https://cdn.intechopen.com/books/images_new/9644.jpg",editedByType:"Edited by",editors:[{id:"51959",title:"Dr.",name:"Masaki",middleName:null,surname:"Kanao",slug:"masaki-kanao",fullName:"Masaki Kanao"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"10432",title:"Casting Processes and Modelling of Metallic Materials",subtitle:null,isOpenForSubmission:!1,hash:"2c5c9df938666bf5d1797727db203a6d",slug:"casting-processes-and-modelling-of-metallic-materials",bookSignature:"Zakaria Abdallah and Nada Aldoumani",coverURL:"https://cdn.intechopen.com/books/images_new/10432.jpg",editedByType:"Edited by",editors:[{id:"201670",title:"Dr.",name:"Zak",middleName:null,surname:"Abdallah",slug:"zak-abdallah",fullName:"Zak Abdallah"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9671",title:"Macrophages",subtitle:null,isOpenForSubmission:!1,hash:"03b00fdc5f24b71d1ecdfd75076bfde6",slug:"macrophages",bookSignature:"Hridayesh Prakash",coverURL:"https://cdn.intechopen.com/books/images_new/9671.jpg",editedByType:"Edited by",editors:[{id:"287184",title:"Dr.",name:"Hridayesh",middleName:null,surname:"Prakash",slug:"hridayesh-prakash",fullName:"Hridayesh Prakash"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8415",title:"Extremophilic Microbes and Metabolites",subtitle:"Diversity, Bioprospecting and Biotechnological Applications",isOpenForSubmission:!1,hash:"93e0321bc93b89ff73730157738f8f97",slug:"extremophilic-microbes-and-metabolites-diversity-bioprospecting-and-biotechnological-applications",bookSignature:"Afef Najjari, Ameur Cherif, Haïtham Sghaier and Hadda Imene Ouzari",coverURL:"https://cdn.intechopen.com/books/images_new/8415.jpg",editedByType:"Edited by",editors:[{id:"196823",title:"Dr.",name:"Afef",middleName:null,surname:"Najjari",slug:"afef-najjari",fullName:"Afef Najjari"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9731",title:"Oxidoreductase",subtitle:null,isOpenForSubmission:!1,hash:"852e6f862c85fc3adecdbaf822e64e6e",slug:"oxidoreductase",bookSignature:"Mahmoud Ahmed Mansour",coverURL:"https://cdn.intechopen.com/books/images_new/9731.jpg",editedByType:"Edited by",editors:[{id:"224662",title:"Prof.",name:"Mahmoud Ahmed",middleName:null,surname:"Mansour",slug:"mahmoud-ahmed-mansour",fullName:"Mahmoud Ahmed Mansour"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},subject:{topic:{id:"802",title:"Operations Management",slug:"industrial-engineering-and-management-operations-management",parent:{title:"Industrial Engineering and Management",slug:"industrial-engineering-and-management"},numberOfBooks:1,numberOfAuthorsAndEditors:1,numberOfWosCitations:2,numberOfCrossrefCitations:7,numberOfDimensionsCitations:10,videoUrl:null,fallbackUrl:null,description:null},booksByTopicFilter:{topicSlug:"industrial-engineering-and-management-operations-management",sort:"-publishedDate",limit:12,offset:0},booksByTopicCollection:[{type:"book",id:"3723",title:"Management and Services",subtitle:null,isOpenForSubmission:!1,hash:"fd3d170b6b6bfc78a9568d26c89ca435",slug:"management-and-services",bookSignature:"Mamun Habib",coverURL:"https://cdn.intechopen.com/books/images_new/3723.jpg",editedByType:"Edited by",editors:[{id:"12501",title:"Prof.",name:"Dr. Md. Mamun",middleName:null,surname:"Habib",slug:"dr.-md.-mamun-habib",fullName:"Dr. Md. Mamun Habib"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],booksByTopicTotal:1,mostCitedChapters:[{id:"11653",doi:"10.5772/9950",title:"An Empirical Research of ITESCM (Integrated Tertiary Educational Supply Chain Management) Model",slug:"an-empirical-research-of-itescm-integrated-tertiary-educational-supply-chain-management-model",totalDownloads:3047,totalCrossrefCites:7,totalDimensionsCites:9,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Mamun Habib",authors:null},{id:"11655",doi:"10.5772/9952",title:"Nonfunctional Requirements Validation Using Nash Equilibria",slug:"nonfunctional-requirements-validation-using-nash-equilibria",totalDownloads:2026,totalCrossrefCites:0,totalDimensionsCites:1,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Andreas Gregoriades and Vicky Papadopoulou",authors:null},{id:"11654",doi:"10.5772/9951",title:"Learning 2.0: Collaborative Technologies Reshaping Learning Pathways",slug:"learning-2-0-collaborative-technologies-reshaping-learning-pathways",totalDownloads:1493,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Veronica Popovici and Ramona Nicoleta Bunda",authors:null}],mostDownloadedChaptersLast30Days:[{id:"11653",title:"An Empirical Research of ITESCM (Integrated Tertiary Educational Supply Chain Management) Model",slug:"an-empirical-research-of-itescm-integrated-tertiary-educational-supply-chain-management-model",totalDownloads:3046,totalCrossrefCites:7,totalDimensionsCites:9,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Mamun Habib",authors:null},{id:"11657",title:"Realization of Lowpass and Bandpass Leapfrog Filters Using OAs and CCCIIs",slug:"realization-of-lowpass-and-bandpass-leapfrog-filters-using-oas-and-ccciis",totalDownloads:3632,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Yanhui Xi and Hui Peng",authors:null},{id:"11655",title:"Nonfunctional Requirements Validation Using Nash Equilibria",slug:"nonfunctional-requirements-validation-using-nash-equilibria",totalDownloads:2026,totalCrossrefCites:0,totalDimensionsCites:1,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Andreas Gregoriades and Vicky Papadopoulou",authors:null},{id:"11654",title:"Learning 2.0: Collaborative Technologies Reshaping Learning Pathways",slug:"learning-2-0-collaborative-technologies-reshaping-learning-pathways",totalDownloads:1493,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Veronica Popovici and Ramona Nicoleta Bunda",authors:null},{id:"11656",title:"Constructing Geo-Information Sharing GRID Architecture",slug:"constructing-geo-information-sharing-grid-architecture",totalDownloads:1561,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:"management-and-services",title:"Management and Services",fullTitle:"Management and Services"},signatures:"Qiang Liu and Boyan Cheng",authors:null}],onlineFirstChaptersFilter:{topicSlug:"industrial-engineering-and-management-operations-management",limit:3,offset:0},onlineFirstChaptersCollection:[],onlineFirstChaptersTotal:0},preDownload:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[{type:"book",id:"10176",title:"Microgrids and Local Energy Systems",subtitle:null,isOpenForSubmission:!0,hash:"c32b4a5351a88f263074b0d0ca813a9c",slug:null,bookSignature:"Prof. Nick Jenkins",coverURL:"https://cdn.intechopen.com/books/images_new/10176.jpg",editedByType:null,editors:[{id:"55219",title:"Prof.",name:"Nick",middleName:null,surname:"Jenkins",slug:"nick-jenkins",fullName:"Nick Jenkins"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter"}}],offset:8,limit:8,total:1},route:{name:"profile.detail",path:"/profiles/173132/farhad-aghdasi",hash:"",query:{},params:{id:"173132",slug:"farhad-aghdasi"},fullPath:"/profiles/173132/farhad-aghdasi",meta:{},from:{name:null,path:"/",hash:"",query:{},params:{},fullPath:"/",meta:{}}}},function(){var e;(e=document.currentScript||document.scripts[document.scripts.length-1]).parentNode.removeChild(e)}()