\r\n\t \r\n\tRecently in 2019, International Council on Systems Engineering (INCOSE) has released the latest version of the “Guidelines for the Utilization of ISO/IEC/IEEE 15288 in the Context of System of Systems (SoS) Engineering” to industry for review and comments. The document was developed under the Partner Standards Development Organization cooperation agreement between ISO and IEEE, as it was approved by Council Resolution 49/2007. This document provides guidance for the utilization of ISO/IEC/IEEE 15288 in the context of SoS in many domains, including healthcare, transportation, energy, defense, corporations, cities, and governments. This document treats an SoS as a system whose elements are managerially and/or operationally independent systems, and which together usually produce results that cannot be achieved by the individual systems alone. This INCOSE guide book perceives that SoS engineering demands a balance between linear procedural procedures for systematic activity and holistic nonlinear procedures due to additional complexity from SoS perspectives. \r\n\tThe objective of this book is to provide a comprehensive reference on Systems-of-Systems Engineering, Modeling, Simulation and Analysis (MS&A) for engineers and researchers in both system engineering and advanced mathematical modeling fields. \r\n\tThe book is organized in two parts, namely Part I and Part II. Part I presents an overview of SOS, SOS Engineering, SOS Enterprise Architecture (SOSEA) and SOS Enterprise (SOSE) Concept of Operations (CONOPS). Part II discusses SOSE MS&A approaches for assessing SOS Enterprise CONOPS (SOSE-CONOPS) and characterizing SOSE performance behavior. Part II focuses on advanced mathematical application concepts to address future complex space SOS challenges that require interdisciplinary research involving game theory, probability and statistics, non-linear programming and mathematical modeling components.
\r\n
\r\n\tPart I should include topics related to the following areas: \r\n\t- SOS and SOS Engineering Introduction \r\n\t- Taxonomy of SOS \r\n\t- SOS Enterprise (SOSE), SOSE CONOPS, Architecture Frameworks and Decision Support Tools
\r\n
\r\n\tPart II should address the following research areas: \r\n\t- SOS Modeling, Simulation & Analysis (SOS M&SA) Methods \r\n\t- SOS Enterprise Architecture Design Frameworks and Decision Support Tools \r\n\t- SOS Enterprise CONOPS Assessment Frameworks and Decision Support Tools.
",isbn:null,printIsbn:null,pdfIsbn:null,doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,hash:"677fbbd5fc2550e8be540f40c0969a62",bookSignature:"Dr. Tien Manh Nguyen",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/7772.jpg",keywords:"Autonomy of Constituents, Operational Independence, Taxonomy, Acquisition Classification, SOS Enterprise Architecture Design, Decision Support Tools, State Modeling, SOS Simulation Methods, DOD Architecture Framework, Enterprise System Engineering, SOS Enterprise CONOPS, Satellite Operations (SATOPS)",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"October 31st 2019",dateEndSecondStepPublish:"November 21st 2019",dateEndThirdStepPublish:"January 20th 2020",dateEndFourthStepPublish:"April 9th 2020",dateEndFifthStepPublish:"June 8th 2020",remainingDaysToSecondStep:"15 days",secondStepPassed:!0,currentStepOfPublishingProcess:3,editedByType:null,kuFlag:!1,editors:[{id:"210657",title:"Dr.",name:"Tien",middleName:"Manh",surname:"Nguyen",slug:"tien-nguyen",fullName:"Tien Nguyen",profilePictureURL:"https://mts.intechopen.com/storage/users/210657/images/system/210657.jpg",biography:"Dr. Tien M. Nguyen received his M.A. in Mathematics, and his Ph.D. in Applied Mathematics from the Claremont Graduate University; M.S.E.E. in Communication Systems Theory from University of California San Diego; and B.S.E. in Electronics and M.S.E. in Electromagnetic Field Theory from California State University Fullerton (CSUF). He also completed all course requirements and passed the comprehensive exam for his M.S.E.E. in Digital Signal Processing from California State University Long Beach. Dr. Nguyen is an expert in Satellite Operations (SATOPS), Satellite Communications (SATCOMs), advanced mathematical modeling for complex systems-of-systems, sensing and communication networks.\nCurrently, he serves as Adjunct Research Professor at CSUF, Mathematics Dept. Concurrently, he is also with the Aerospace Corporation, serving as a Sr. P.E. in Space System Group Program Office, Space System Architect Division, Global Partnerships Subdivision. He has more than 13-years of service at Aerospace, and prior to his current position; he has served as Sr. Engineering Specialist, Sr. Project Lead, Section Manager, Associate Director, Interim Director, and Principal Technical Staff (the highest technical level at the corporation). At Aerospace, he invented HPA linearizer, GMSK synchronizers and developed advanced optimization techniques using game theory for achieving affordable and low-risk acquisition strategy. Prior to CSUF, he had also held a Research Assistant Professor at the Catholic University of America in concurrent with The Aerospace Corporation positions. \nHe was a Engineering Fellow from Raytheon, where he had 10-year of services at Raytheon, serving as Program Area Chief Engineer, Program Chief Engineer, PI, Technical Director, Program Manager, Lead Architect and Lead System Engineer for many advanced programs and pursuits related to sensing and communication networks. At Raytheon, he invented radar-communication technology and gun barrel detector using millimeter-wave. Previous to Raytheon and Aerospace Corporation, Dr. Nguyen was with NASA/JPL for more than 11-years, where he served as the NASA delegate to the international Consultative Committee for Space Data System (CCSDS). Many of his works on RF and Modulation were adopted as the CCSDS standards for USB waveforms and space RF systems. At JPL he invented QPSK phase ambiguity resolver and developed innovative optimization technique for simultaneous range-command-telemetry operation. He built the first laser lab and automated manufacturing lab when he was with ITT Technical Services in the early ’80s. \nHe has published more than 250 technical reports and papers. His work has appeared in NASA TechBrief, textbook, Open Access Book, SIAM Publication, CCSDS Blue Book, and Wiley & Sons Encyclopedia of Electrical and Electronics Engineering. He was selected as a Vietnamese-American Role Model by KCSI-TV, Channel 18 in 2002, and Recognition Honoree at 50-Year Celebration of CSUF in 2007. He received numerous Raytheon, Aerospace and NASA awards, and Air Force commendations. He holds 16 patents. His biography has been listed in Marquis Who’sWho in Science and Engineering in America.",institutionString:"The Aerospace Corporation",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"California State University, Fullerton",institutionURL:null,country:{name:"United States of America"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"11",title:"Engineering",slug:"engineering"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"305835",firstName:"Ketrin",lastName:"Polesak",middleName:null,title:"Mrs.",imageUrl:"https://mts.intechopen.com/storage/users/305835/images/9351_n.png",email:"ketrin@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review, to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. Whether that be identifying an exceptional author and proposing an editorship collaboration, or contacting researchers who would like the opportunity to work with IntechOpen, I establish and help manage author and editor acquisition and contact."}},relatedBooks:[{type:"book",id:"3161",title:"Frontiers in Guided Wave Optics and Optoelectronics",subtitle:null,isOpenForSubmission:!1,hash:"deb44e9c99f82bbce1083abea743146c",slug:"frontiers-in-guided-wave-optics-and-optoelectronics",bookSignature:"Bishnu Pal",coverURL:"https://cdn.intechopen.com/books/images_new/3161.jpg",editedByType:"Edited by",editors:[{id:"4782",title:"Prof.",name:"Bishnu",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"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"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1373",title:"Ionic Liquids",subtitle:"Applications and Perspectives",isOpenForSubmission:!1,hash:"5e9ae5ae9167cde4b344e499a792c41c",slug:"ionic-liquids-applications-and-perspectives",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/1373.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"4816",title:"Face Recognition",subtitle:null,isOpenForSubmission:!1,hash:"146063b5359146b7718ea86bad47c8eb",slug:"face_recognition",bookSignature:"Kresimir Delac and Mislav Grgic",coverURL:"https://cdn.intechopen.com/books/images_new/4816.jpg",editedByType:"Edited by",editors:[{id:"528",title:"Dr.",name:"Kresimir",surname:"Delac",slug:"kresimir-delac",fullName:"Kresimir Delac"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"57",title:"Physics and Applications of Graphene",subtitle:"Experiments",isOpenForSubmission:!1,hash:"0e6622a71cf4f02f45bfdd5691e1189a",slug:"physics-and-applications-of-graphene-experiments",bookSignature:"Sergey Mikhailov",coverURL:"https://cdn.intechopen.com/books/images_new/57.jpg",editedByType:"Edited by",editors:[{id:"16042",title:"Dr.",name:"Sergey",surname:"Mikhailov",slug:"sergey-mikhailov",fullName:"Sergey Mikhailov"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3794",title:"Swarm Intelligence",subtitle:"Focus on Ant and Particle Swarm Optimization",isOpenForSubmission:!1,hash:"5332a71035a274ecbf1c308df633a8ed",slug:"swarm_intelligence_focus_on_ant_and_particle_swarm_optimization",bookSignature:"Felix T.S. Chan and Manoj Kumar Tiwari",coverURL:"https://cdn.intechopen.com/books/images_new/3794.jpg",editedByType:"Edited by",editors:[{id:"252210",title:"Dr.",name:"Felix",surname:"Chan",slug:"felix-chan",fullName:"Felix Chan"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3621",title:"Silver Nanoparticles",subtitle:null,isOpenForSubmission:!1,hash:null,slug:"silver-nanoparticles",bookSignature:"David Pozo Perez",coverURL:"https://cdn.intechopen.com/books/images_new/3621.jpg",editedByType:"Edited by",editors:[{id:"6667",title:"Dr.",name:"David",surname:"Pozo",slug:"david-pozo",fullName:"David Pozo"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"371",title:"Abiotic Stress in Plants",subtitle:"Mechanisms and Adaptations",isOpenForSubmission:!1,hash:"588466f487e307619849d72389178a74",slug:"abiotic-stress-in-plants-mechanisms-and-adaptations",bookSignature:"Arun Shanker and B. Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"51617",title:"Herpes Simplex Virus 1 and 2 Vaccine Design: What can we Learn from the Past?",doi:"10.5772/64447",slug:"herpes-simplex-virus-1-and-2-vaccine-design-what-can-we-learn-from-the-past-",body:'\n
\n
1. Introduction
\n
This review will be devoted to the topics of not yet marketed HSV vaccine(s), which design is still in the focus of interest, especially from the point of view of immunotherapeutic use. The human herpesvirus 1 (common name herpes simplex virus 1, HSV-1) is one of the first human viruses discovered [1] and belongs among the most intensively investigated viruses. In the span of the last 45 years, the PubMed portal has registered nearly 11,000 papers devoted to various aspects of HSV-1/HSV-2, starting from virus structure, continued by the molecular mechanisms of lytic replication versus latency maintenance, through the virus spread in the body and the eliciting of different forms of immune response, not omitting many clinical and epidemiological studies. In the second half of the last century, two subtypes of herpes simplex virus (HSV) have been described [2] and were designated HSV type 1 and HSV type 2 [3]. While the former (HSV type 1) has been predominantly isolated from the orofacial area and upper respiratory airways, the latter was believed to infect the urogenital tract and occasionally the newborn. According to recent classification, the HSV-2 represents a distinct species of the Simplexvirus genus, which along with the Varicellovirus genus belongs to subfamily Herpesvirinae of the Herpesviridae family. Nevertheless, both species are closely related, since they differ only in a few antigenic domains (and/or epitopes) located in the envelope glycoproteins, namely, in an entirely distinct gG and/or in the partially unrelated gC [4].
\n
\n
\n
2. The molecular biology of HSV: virion structure, lytic replication and latency
\n
The HSV virion consists of four elements: (a) a core containing the viral DNA (vDNA), (b) an icosahedral capsid surrounding the core, (c) an unstructured proteinaceous layer called the tegument that surrounds the capsid, and (d) an outer lipid bilayer envelope exhibiting spikes on its surface. The core contains the double-stranded (ds) DNA (dsDNA) genome wrapped as a toroid or spool in a liquid crystalline state. A small fraction of the virion DNA may be circular, but the bulk of packaged HSV DNA is linear and double-stranded. The enveloped virion is a spherical particle with an average diameter of 186 nm, which might extend to 225 nm with the spikes included, while the internal capsid has a constant diameter of 110 nm (reviewed at [5]). The HSV virion contains more than 30 proteins that were designated as virion polypeptides (VPs). Out of these approximately known and additional 10 suspected virion proteins (VPs), at least 11 are located within the envelope at the surface of virion (predominantly accessible to antibody), from which at least 10 are glycosylated (reviewed at [6]).
\n
The HSV genes were classified into at least three general kinetic classes: alpha or immediate-early (IE), beta or early (E), and gamma or late (L) genes. The IE mRNAs are transcribed by the help of a transcription-inducing cofactor present in the virion tegument, also called alpha-TIF/VP16. In brief, the tegument protein VP16 is becoming a part of a tripartite complex comprised of octamer-binding protein 1 (Oct-1) and host cell factor 1 (HCF-1). At least two of the IE proteins (ICP0 and ICP4; Figure 1) are transactivators needed for initiation of transcription of the E/beta mRNAs (the first checkpoint) and later also for transcription of gamma/L mRNAs. The latter are mainly structural virion proteins in contrast to E transcripts specifying nonstructural polypeptides. The structural proteins are transcribed in two waves, called gamma1/L and gamma2/L. The former occurs more closely related with the E transcription phase (second checkpoint) and requires predominantly ICP4. The expression of beta polypeptides as well as that of group gamma1/L genes requires ICP4 and at low multiplicities of infection also ICP0. The expression of gamma2/L genes, controlled at the third checkpoint, requires ongoing DNA synthesis as well as the presence of ICP4 and ICP0 [7].
\n
Figure 1.
The IE genes and positions of their ORFs in the HSV genome.
\n
The HSV genome consists of two covalently linked sequence components, designated as L (long) and S (short). Each component consists of unique (U) sequences bracketed by inverted repeats (IR) [6].
\n
The entry of HSV into cells involves the interaction of at least five virion surface glycoproteins (gB, gC, gD, gH and gL) with several receptors on the cell surface following fusion of the envelope with a cellular membrane. The HSV glycoproteins along with other proteins important for pathogenicity are listed in Table 1. The first step in the process of entry is the binding of virions to glycosaminoglycans (GAGs) on the cell surface. The interaction of virions with GAGs is mediated by two glycoproteins: gC and gB. The next step in virus entry consists of the interaction of gD with its receptors and execution of fusion between the envelope and the cellular membrane by the heterodimer gH/gL and also by gB. The interaction of gD with its receptors has been extensively studied in several laboratories. The consensus is that, for entry, gD interacts with one of three natural receptors (reviewed at [8]). The receptors are nectins, a protein-designated herpesvirus entry mediator (HVEM) and a selected form of 3-O-sulfated heparan sulfate (3-OS HS). Nectins are intracellular adhesion molecules expressed on epithelial and neural cells and are members of the extended immunoglobulin (Ig) family. Earlier studies showed that gD of HSV-1 interacts with the amino-terminal V1 domain of nectin 1. The wild-type HSV-2 can enter susceptible cells via an alternative receptor, namely, nectin 2. HVEM is a member of the extended tumor necrosis factor receptor (TNFR) family, expressed mainly on T lymphocytes but occasionally also on natural killer (NK) cells. Its natural ligand is LIGHT (homologous to lymphotoxins, exhibits inducible expression), which is constitutively expressed on T and natural killer (NK) cells and appears to be a regulator of mucosal immune system. The LIGHT receptor competes with HVEM for the gD (reviewed at [9]). The role of gD with respect to gB and gH/gL is to recruit and position them properly to enable interaction with GAGs and with the lipid bilayers of the cellular membranes [10].
\n
\n
\n
\n
\n\n
\n
Gene
\n
Protein
\n
Function
\n
\n\n\n
\n
RL1
\n
γ34.5
\n
L protein, cofactor for ribosomal translation
\n
\n
\n
RL2
\n
ICP0
\n
IE protein, spliced, three exons, a RING finger motif, acts as a nonspecific transactivator of any beta and gamma gene
\n
\n
\n
UL1
\n
gL
\n
Late glycoprotein, forms the gH/gL complex, syn mutation
\n
\n
\n
UL5
\n
\n
E class protein, part of the helicase/primase complex
\n
\n
\n
UL9
\n
\n
E/L class, ori-binding protein, cooperates with ICP8 at initiation of vDNA replication
\n
\n
\n
UL10
\n
gM
\n
L class, glycosylated polypeptide, interacts with gN, participates in exocytosis and in cell-to-cell spread
\n
\n
\n
UL18
\n
VP24
\n
L class, capsid component, participates in triplex formation
\n
\n
\n
UL19
\n
VP5
\n
L class protein, the major capsid component (149/150K)
\n
\n
\n
UL21
\n
\n
L class tegument protein weakly capsid-associated, binds to microtubules at axonal transport
\n
\n
\n
UL22
\n
gH
\n
L class, glycosylated protein, essential for penetration to cells, needs complexing with gL
\n
\n
\n
UL23
\n
TK
\n
E class, tegument protein, thymidine kinase, virulence factor
\n
\n
\n
UL26
\n
\n
L class, “scaffolding” capsid protein, the N-terminus required for virion assembly
\n
\n
\n
UL27
\n
gB
\n
L class glycoprotein, essential for adsorption and membrane fusion, syn3 locus
\n
\n
\n
UL29
\n
ICP8
\n
E class, ssDNA binding, required for vDNA synthesis, keeping the DNA fork apart
\n
\n
\n
UL30
\n
DNA pol
\n
E/L class protein, DNA polymerase (elongation enzyme), virulence factor
\n
\n
\n
UL35
\n
VP26
\n
L class virion protein, hexon component, also termed NC7
\n
\n
\n
UL36
\n
VP1/ VP2
\n
L class, large tegument protein, important for egress through cytoplasm and re-envelopment
\n
\n
\n
UL38
\n
VP19C
\n
L class capsid protein, triplet component with VP23 (1:2), connects the hexons and pentons
\n
\n
\n
UL39
\n
RR
\n
E class protein, large subunit of RR, membrane-anchored protein kinase
\n
\n
\n
UL40
\n
RR
\n
E class protein, the small subunit of RR
\n
\n
\n
UL41
\n
\nvhs\n
\n
L class, tegument protein, interferes with the host cell proteosynthesis, virulence factor
\n
\n
\n
UL44
\n
gC
\n
L class, glycoprotein, reacts with GAG on cell surface, binds C3 and/or C5, virulence factor
\n
\n
\n
UL48
\n
Alpha- TIF /VP16
\n
L class tegument protein, alpha-transinducing factor
\n
\n
\n
UL49.5
\n
gN
\n
L class membrane-associated small glycoprotein, complexes with gM
\n
\n
\n
UL53
\n
gK
\n
L class glycoprotein (40K), localizes to Golgi and ER, involved in egress, syn1 locus
\n
\n
\n
UL54
\n
ICP27/ IE63
\n
IE class protein, blocks cellular mRNA transport to cytoplasm
\n
\n
\n
RS1
\n
ICP4
\n
IE class protein, transactivator for E and L promoters
\n
\n
\n
US1
\n
ICP22
\n
IE class regulatory protein, essential in animal experiments, cellular cyclin A and B degradation
\n
\n
\n
US4
\n
gG
\n
L class, glycosylated protein and envelope component, HSV-2 specific
\n
\n
\n
US6
\n
gD
\n
E/L class, glycosylated protein, 56K, adsorption to nectin 1 and HVEM protein receptors, essential for entry to cells, neural uptake
\n
\n
\n
US7
\n
gI
\n
L class glycoprotein, forms heterodimer with gE, the complex facilitates cell-to-cell spread, neural uptake
\n
\n
\n
US8
\n
gE
\n
L class glycoprotein, complexes with gI, essential for neural uptake
\n
\n
\n
US12
\n
ICP47
\n
IE class protein, blocks the transport of viral antigenic epitopes
\n
\n
\n
LAT
\n
\n
Three categories of RNAs (8.5 kb, 2.0, and 1.5 kb), the small LATs terminate antisense to ICP0 ORF. Latency regulator
\n
\n\n
Table 1.
HSV-1 genes and corresponding proteins regarded for importance in virulencea.
Abbreviations: vDNA = viral DNA; ICP = infected cell protein; VP = virion protein (structural); IE = immediate-early kinetics (class); E = early kinetics (class); L = late kinetics (class); K = 1,000 kDa (Mr); RR = ribonucleotide reductase; vhs = virion host shutoff; LAT = latency-associated transcript(s); RNA molecules without capping, do not interact with ribosomes; UL = unique long (DNA segment); US = unique short; RL = repeat long; RS = repeat short; HVEM = herpes virus entry mediator)
The gB belongs to class III fusion protein that after GAG interaction facilitates fusion of the virion envelope bilayer with the cellular membrane. A recent study showed that gB can also bind to the paired immunoglobulin-like type 2 receptor-α (PILRα) to trigger viral fusion in certain cell types; however, the precise role of this interaction in viral entry remains to be determined [11]. The gH and gL appear to form a tight complex, since neither protein is stable without the other. In contrast to gB, the structural studies of gH/gL showed no homology with fusion domains of other viral glycoproteins. The exact role of gH/gL has not been determined; however, one hypothesis is that the interaction of gD with its receptors changes the conformation of gH/gL, which in turn induces gB to adopt its fusion conformation [12]. Even that gE had been regarded for not essential in cell culture, it has been shown very important at entering nerve endings (neural uptake) and subsequent neural spread [13].
\n
The latency of HSV can be characterized by circularized state of the HSV genome in the absence of IE and/or E transcription, especially lacking the expression of the two most important beta/gamma transcription activators, namely, ICP0 and ICP4 (reviewed at [14]). We pointed at the possibility that the IE block in question might be “leaky” since small amounts of ICP4 mRNA could be found in the non-cultured ganglion explants which would later on yield infectious virus [15]. Overwhelming literature deals with the hypotheses that the small latency-associated transcripts RNA (LATs), RNA molecules expressed during latency contribute to the silencing of ICP0 mRNA transcription as well as to the maintenance of latency [16]. It has been shown that LAT can reduce the expression of viral genes and suppress HSV replication in cultured cells. In addition, LAT probably protects the HSV DNA carrying neuron from apoptosis. The anti-apoptosis activity of LAT has been independently confirmed in tissue culture and in the mouse ocular model [17]. Further data suggest the participation of LAT in reactivation of the latent genome [18, 19].
\n
\n
\n
3. Pathogenesis of HSV-1 and/or HSV-2 infections
\n
The crucial event in HSV pathogenesis is, when the virus, which has reached the pseudounipolar neuron of the regional sensory ganglion via the quick axonal transmission, may but need not undergo productive replication. Thus, latency can be established from the very beginning, that is, in the absence of any transcription provided by the virus-coded alpha-transinducing factor (alpha-TIF). As described by Efstathiou et al. [20], the latent genome resides in the nuclei of neurons in the form of a circularized nonintegrated plasmid-like structure. This structure survives within the carrier neuron. the carrier neuron, so that neural cells harbor the viral DNA for their lifetime. The tegument-bound alpha-transinducing factor (alpha-TIF) might be “lost” during intra-axonal transport [21]. Some axons might be as long as 500 mm (as a rule about 100 mm), while the minimum speed for HSV capsid transmission is about 1–2 mm/h. The quick axonal transport along neurotubules is provided by the dynein molecule, which binds the capsid component UL21. The environment within sensory neurons favors the establishment of latency rather than productive virus replication also because the neuronal transcription cofactors such as Oct-2 and/or Brn-3 repress the IE promoters [22, 23].
\n
During reactivation, the blocked IE checkpoint of virus replication must be overcome especially by overwhelming expression of ICP0 protein in order to achieve productive virus replication and infectious HSV production [24]. The reactivation process can be hampered by two different mechanisms. One depends on viral genome products interacting with cellular cofactors of transcription within neurons. It may not be excluded that due to the existence of at least two activation checkpoints, controlling the expression gamma1 or E/L proteins (glycoproteins) and the gamma2/L proteins (glycoproteins), also incomplete virus particles may be formed. The E and/or E/L proteins may be synthesized in the absence of complete virion formation, or assembly of virions with defective vDNA occurs. However, the presence of virus-coded antigens (even in the absence of infectious virus particles) could induce a potent immune response (including the accumulation of cytotoxic T cells in the vicinity of HSV carrier neurons). It should be also mentioned that the IE protein ICP47 acts by immune evasion. Alternative effects inhibiting the antiviral defense can be exerted by gC [25], which binds complement, and by the gE molecule, which binds the Fc fragment of immunoglobulins.
\n
If reactivated virus reaches the peripheral tissues by retrograde axonal transport (skin squamous epithelium, non-cornified squamous epithelium of mouth mucous membrane, corneal epithelium), then it starts to replicate and causes inapparent virus shedding or blister formation. A nice example for creating favorable conditions of virus replication at peripheral skin was described in the ear model [26, 27]. These investigators found that prostaglandins produced after skin trauma or UV light irradiation would enhance the replication of recurrent HSV-1. As shown by Walz et al. [28], neurectomy of the trigeminal nerve root reactivated the latent HSV harbored within ganglion cells. The round trip of reactivated virus usually ends with reinfection of additional neurons within the regional sensory ganglion. Less frequently, the retrograde axonal transport may continue in centripetal direction, that is, to brain stem. Furthermore, the central nervous system (CNS) may become infected via bloodstream (in newborn) or along the olfactory route (reviewed at [29]).
\n
The different manifestations of clinical disease reflect the above-described mechanisms of HSV spread in the body. Essentially, HSV-1 can induce the acute primoinfection (gingivostomatitis) and recurrent disease (classical labial herpes). Alternatively, the ocular herpes, often manifested as herpes keratitis, occurs rather as recurrent disease than primoinfection. In newborn as well as in the immunocompromised host and/or in the case of local inflammation in the skin (e.g., due to allergic manifestations such as atopic eczema), the HSV might cause generalized skin disease or even viral sepsis (i.e., the hepatoadrenal necrosis in newborn or severe meningoencephalitis). In newborn, perinatal infection causes an acute dissemination of HSV-2 via bloodstream. Meningitis and meningoencephalitis of neonates, similarly as various forms of genital infection of man and women, are mainly HSV-2-related. In women, the outer but also the inner genital tract (vagina or cervical mucosa covered with non-cornified squamous epithelium) might become infected. In the skin as well as at the mucous membranes covered by squamous epithelium, both HSV-1 and HSV-2 species replicate within the lower and/or medium squamous cell layers causing blister formation [30].
\n
\n
\n
4. Mechanisms of the immune response to HSV-1 or HSV-2
\n
The immune response to HSV-1 and/or HSV-2 is induced in an early and a late phase. During early phase, the nonspecific antiviral mechanisms are activated, while in the late phase, the HSV-specific reaction is mobilized. The specific immune response culminates by recruiting the CD8+ cytotoxic T lymphocytes important for extracellular latency control, in contrast to the intracellular latency control which has been discussed above. The specific cell-mediated immune response may begin in the regional lymph nodes draining the virus inoculation site (both T-lymphocyte lines are engaged) as well as in the trigeminal ganglia which harbor the latent virus (mainly cytotoxic CD8+ T cells take part).
\n
\n
4.1. The innate immune response
\n
The innate antiviral immunity to HSV-1 and/or HSV-2 is provided by alpha/beta type I interferons (IFNs) as well as by activation of natural killer (NK) cells. The role of type I IFNs in anti-HSV defense was demonstrated in knockout mice deficient in type I IFN production [31, 32]. The type I IFN release is induced by virus constituents called pathogen-associated molecular patterns (PAMPs) interacting with the corresponding pattern recognition receptors (PRRs) at the surface of responding cells. The best known examples are the Toll-like receptors (TLR). For example, the envelope glycoprotein D molecule of the HSV-1 interacts with the TLR-2 receptor on the target cell membrane [33], while the TLR-9 receptor binds to non-methylated CpG motifs of the vDNA present in the nuclei of infected cells [34]. It has been also shown that children with TLR-3 defects are more susceptible to herpes encephalitis [35, 36]. Both types I of IFNs act on the receptors in neighboring cells eliciting the synthesis of antiviral substances such as the 2′,5′-oligoadenylate synthase, RNase L, and/or RNA-dependent protein kinase. The type I IFN also inhibits cell proliferation and enhances the activation of NK cells as well as the expression of MHC class I molecules, but downregulates MHC class II expression. The type III IFNs, also designated as IFN-λ1, IFN-λ2 and IFN-λ3 (IL-29, IL-28A, and IL-28B), have been recently characterized; alike to classical type I IFNs, also IFN-λ induces production of antiviral substances in the infected cells [37]. The participation of type III IFNs in inhibition of HSV replication has been confirmed in vitro [38, 39].
\n
In addition to type I IFN formation, the activation of NK cells represents another important arm of nonspecific immune response to HSV-1 and HSV-2 infection [40]. The nonspecific killers belong to the group of class I innate lymphoid cells. Morphologically, they belong to large granular lymphocytes (LGLs), which cytoplasm contains azurophilic granules with perforins and granzymes A and B. They exert cytolytic activities helpful at the elimination of infected cells. The perforins create a transmembrane channel piercing the cell membrane, while granzymes activate the apoptosis of target cells. The NK cells also produce IFN-gamma, which promotes their activation along with the activation of macrophages. In association with this, enhanced death rates were noted in NK-deficient mice infected with HSV [41].
\n
The elimination of HSV-1 and/or HSV-2 infected cells at the portal of entry is mediated by additional immune cells such as neutrophils, dendritic cells (DCs), and macrophages. The neutrophils secrete tumor necrosis factor (TNF) which induces apoptosis of infected cells via caspase-8 activation [42]. Blood monocytes when entering the connective and/or other tissues are altered into macrophages able to engulf the extracellular virus particles but also residues of apoptotic cells. The active macrophages belong to the group of antigen-presenting cells (APCs) that present the external immunogenic peptides to T lymphocytes. This process called antigen presentation launches the specific immune response. Activated macrophages also release several pro-inflammatory cytokines like TNF and IL-6 (participates in B-cell activation), type I IFNs, and several chemokines such as RANTES regulated upon activation normal T-cell expressed and secreted. Finally, nitric oxide (NO) and other substances are produced by macrophages. It has been demonstrated in experiments in vitro that the replication of HSV-1 in infected cells may be considerably inhibited in the presence of NO released from macrophages [43]. It should be mentioned here that IFN-gamma (belongs to type II IFNs) is in fact a cytokine released from activated NK cells but also produced by the helper CD4+ T lymphocytes as well as by cytotoxic CD8+ T lymphocytes [44]. The IFN-gamma considerably promotes function of activating macrophages and NK cells, in which it induces a production of cytolytic substances. Taken together, both NK cells and macrophages represent an important first-line defense at the early stage of HSV infection, that is, before the onset of specific immune response.
\n
As already mentioned above, important immune cells participating in the initiation of antigen processing are the dendritic cells (DCs). They represent a relatively heterogenous cell population, from which the most active in antiviral defense are the myeloid (conventional) and the plasmacytoid DCs (pDCs), both derived from the myeloid progenitor cells. The myeloid DCs act as antigen-presenting cells (APCs), which carry the processed antigenic peptides into lymph nodes, where the peptides in question are presented to the T lymphocytes. Based on this presentation event, the specific immune response is induced (activation of T and B cells). In DC-depleted mice infected with HSV-1, encephalitis developed with a significantly higher frequency as compared to conventional mice [45]. The plasmacytoid DCs under physiological conditions participate in the development of peripheral immune tolerance. Upon HSV infection, plasmacytoid DCs start to produce high amounts of IFN-alpha, which not only inhibits the virus replication in the otherwise surrounding susceptible cells but also activates lymphocytes and additional DCs. The HSV dsDNA binds to TLR-9 of the plasmacytoid DC which initiates IFN-alpha production [46].
\n
\n
\n
4.2. The acquired immune response
\n
The acquired immune response encounters the activation of both T-lymphocyte lines, namely, those differentiated either in direction of CD4+ or CD8+ T cells. The acquired immune response then is triggered by the activation of T-cell receptors (TCRs), which recognize the HLA I/CD8 and/or the HLA II/CD4 bound antigenic peptides. While the former T cells are involved in the destruction and elimination of HSV-infected cells, the latter acts as helper T cells at inducing the specific antibody production by B cells. At primary infection, the HSV-1 or HSV-2 particles are engulfed by DCs, which move the viral peptides to the regional lymph nodes, where the naïve (unprimed) lymphocytes get first stimulated. The presentation of exogenous viral peptides is achieved by means of the HLA class II molecules, which are recognized by the TCR of the CD4+ T lymphocytes. In contrast, the immunogenic peptides of the newly (de novo) synthesized HSV proteins are presented by HLA class I molecules at the surface of infected cells where they interact with the TCR of cytotoxic CD8+ T lymphocytes [47]. In certain extent, the so-called cross presentation occurs, at which the exogenous viral antigens are binding into the HLA class I molecules that are recognized by the CD8+ T lymphocytes [48]. The activated helper CD4+ T cells release cytokines such as IL-2 (inducing T-lymphocyte proliferation) and IL-4, IL-5 and IL-6 (promoting the differentiation of B cells into plasma cells).
\n
The plasma cells synthesize the specific virus-neutralizing antibodies reacting mainly with the envelope glycoproteins such as gB and gD. The antibodies may also activate complement; on the other hand, the gC molecule of the HSV-1 envelope binds C3b and in less extent also C5b reducing the availability of complement components for virus neutralization. The low-pathogenic HSV-1 gC minus strains may become pathogenic as was confirmed in a C3 knockout murine model [49]. The HSV-1 antibodies belong to IgM as well as IgG class; the HSV-2 antibodies may be also of the secretory IgA class; the latter participates in virus clearance at the genital tract mucosa [50]. Even though the serum antibodies are of importance at the acute phase of infection, they may not fully eliminate the HSV since it invades the nerve endings and spreads to regional sensory ganglia before the development of antibody response. Some studies have shown that passive immunization with immune serum did not prevent latency after genital infection with HSV-2 [51, 52, 53]. Since the latent HSV (episomal vDNA) is frequently harbored in neural tissue for lifelong, the presence of HSV antibodies in the serum of healthy subjects may be interpreted as infectious immunity.
\n
The cytotoxic CD8+ T lymphocytes represent the mainstream of the specific immune response providing clearance of HSV-1- or HSV-2-infected cells from the body. The activation of cytotoxic CD8+ T cells takes place in the regional lymph nodes by means of viral antigenic peptides presented by HLA class I molecules expressed at the surface of DCs. The IL-2 released from helper T cells acts on the precursor cytotoxic T lymphocytes which differentiate into mature cytotoxic CD8+ T cells. These mature CD8+ T cells accumulate at the peripheral virus inoculation site, where they eliminate the HSV-infected cells. The activated cytotoxic T lymphocytes release substances such as perforins, granzyme and granulysin, which destroy the infected target cell and/or induce apoptosis acting on their FAS receptors. They also release cytokines such as IFN-gamma and the tumor necrosis factor (TNF). IFN-gamma has a multiple effect, since it enhances the expression of HLA class I as well as HLA class II molecules [54] and also induces the expression of antiviral substances such as protein kinase R (PKR). This substance causes inhibition of translation of many viral but also cellular proteins. The increased TNF production also enhances the number and activity of HLA class I molecules.
\n
The clearance of HSV-2 from peripheral tissues such as genital mucosa may be also provided by CD4+ T cells. Transgenic mice, which revealed defects in their CD8+ T-lymphocyte activity, have been still well protected against a lethal dose of HSV-2. In contrast, mice with depleted CD4+ T lymphocytes showed slower clearance and less protection against HSV-2 [55–57]. It can be concluded that the clearance of HSV-2 from genital mucosa requires cooperation of both the helper and cytotoxic T lymphocytes. Both T-cell populations can be found in the skin, in the genital and oral mucosa, in the ocular tissue, and also in the trigeminal ganglia surrounding the infected neurons [58–60].
\n
After healing of the acute phase of HSV infection, about 0.1–1% of memory T cells, which may remain within the circulation, are still able to recognize the HSV antigenic epitopes. These epitopes being recognized by CD4+ and/or CD8+ lymphocytes were not all exactly mapped yet, but until now, 22 of them were defined. The HSV-specific epitopes can be found mainly among VPs (structural envelope glycoproteins, tegument proteins and capsid polypeptides), but are also present on the ICPs, that is, on nonstructural enzymes and regulatory proteins. The CD4+ T lymphocytes generated against the HSV-2-specified structural proteins recognize the epitopes of the RR1/UL39 polypeptide, the epitopes of UL46 tegument protein, and several ones on glycoproteins gD and gB [61].
\n
The CD8+ T lymphocytes, surrounding the neurons of the regional sensory ganglia of mice, rabbits, and humans in which viral latency had been established [62–64], are believed to control reactivation [65]. It was demonstrated, for example, that the ICP4/145K IE protein can be digested by granzyme B, which would prevent virus reactivation [66]. The accumulated CD8+ T cells produce IFN-gamma as well, inhibiting the expression of ICP0/110K IE protein [67].
\n
The most intriguing still unexplained question is why do the CD8+ T lymphocytes accumulated in the neighborhood of latent HSV-harboring neurons, since the expression of IE and E polypeptides is widely hampered in them. Interestingly enough, the great majority of CD8+ T lymphocytes in the trigeminal ganglion of mice in which latency had been established possess a TCR which reacts with gB epitope at aa 498–505, while a smaller proportion of them may be stimulated with RR1 epitope aa 822–829. The reactivity of the other T cells was not identified [68, 69]. In human trigeminal ganglia, both the CD4+ and CD8+ T cells were described that react with many antigenic epitopes present in the IE polypeptides ICP0 and ICP4, in the E polypeptides such as thymidine kinase (TK) and RRl, as well as with antigenic domains from structural proteins such as the tegument proteins VP11/12 and VP13/14 and envelope glycoproteins gB, gK, and gL, even though an exact identification of all antigenic epitopes in the HSV proteins was not done yet [70, 71].
\n
The phenotype of CD8+ T lymphocytes which can accumulate within the ganglia was not characterized. The permanently present cytotoxic T lymphocytes might belong to the population of so-called tissue-resident memory T lymphocytes (TRM) or to the population referred as T-effector memory cells (TEM cells). Additional markers such as CD44, CD69, CD62L, and CCR7 were described at their surface, but their full definition is still the matter of investigation [72].
\n
\n
\n
\n
5. Survey of experimental HSV-1 and HSV-2 vaccines and their design
\n
Though the first vaccines against HSV-1 and/or HSV-2 were prepared in the sixties of last century [73], no accepted and fully efficient HSV vaccine is available till now. The main problem is the incomplete understanding of the role of cytotoxic CD8+ T cells in the maintenance and/or of elimination neurons carrying the silenced HSV DNA. Another serious problem is the production of immune evasion polypeptides (reviewed at [74]). The optimal HSV vaccine should overcome the ICP47-mediated interference with TAP, which transports the viral peptides inside ER to bind with HLA molecules. Finally, the last but not negligible problem is that at least some of over 20 immunogenic epitopes, which are distributed within about 80 HSV proteins (both structural and nonstructural), should participate in the vaccine but in an optimal composition. An effective HSV vaccine might differ depending of its purpose, which could be either preventive or immunotherapeutic. An ideal vaccine should not allow the acute disease to develop, and it should be able to prevent the establishment of latency. These goals, especially the latter, are difficult to achieve. The minimum effect provided by an immunotherapeutic vaccine would be to minimize the extent of recurrent lesion originating from the reactivation of the latent viral genome. As described above, the cytotoxic T lymphocytes accumulating in vicinity of HSV DNA carrier cells in the regional sensory ganglion seem of great importance when keeping a state of equilibrium probably inhibiting the transcription of IE genes by an unknown mechanism. One could estimate that in the case of the inefficiency and/or complete failure of the function of cytotoxic T-cell virus, reactivation and subsequent retrograde axonal transport of virions occur. In this section, we describe the various HSV vaccines which have been used under experimental conditions, while the next paragraph will describe their efficacy in human volunteers.
\n
\n
5.1. Inactivated virion and subunit vaccines
\n
The inactivated HSV-1 and/or HSV-2 vaccines consist of infected cell extracts, which have been introduced in the 1980s of last century, partially purified and inactivated either by UV light, by thermal treatment, or with formalin [75–77]. The efficacy of these first vaccines was tested mainly in mice, but some of them were used also in human trials (see Section 6). Especially in animal models, a good protective effect against acute virus challenge has been found not to prevent the establishment of latency. However, when testing the effect of a virion-free HSV vaccine prepared by extraction of infected LEP cells with Nonidet P-40 [78] in the rabbit corneal model, Rajčáni et al. [79] concluded that even though vaccination prior to infection did not fully prevent latency, it considerably reduced the number of regional sensory (trigeminal) ganglion cells (neurons), which became the HSV genome carriers. Recently, a formalin-inactivated HSV-2 (FI-HSV2) vaccine mixed with Al(OH)3 adjuvant was tested in combination with monophosphoryl lipid A (MPL) [80]. The latter authors showed that the vaccine protected mice against local vaginal challenge with HSV-2 as well as reduced the extent of latency established in the sensory ganglia of lumbosacral nerve roots.
\n
The subunit vaccines were mainly glycoprotein mixes purified on lectins (e.g., lentil lectin) showing high affinity to the HSV virion envelope antigens. The dominant protective antigens among the 11 HSV glycoproteins are gB and gD, which possess important immunogenic epitopes. The abovementioned glycoproteins elicit virus-neutralizing antibodies, antibodies participating in the antibody-dependent cellular cytotoxic (ADCC) response, and they also activate the T lymphocytes. Several reports described the efficacy of the subunit vaccines of various purity based on either HSV-1 and/or HSV-2 envelope glycoproteins [81–84]. For example, the subunit HSV-1 vaccine (strain HSZP Immuno) had been prepared from chick embryo cells infected with the low-virulent HSZP strain [85]. Tested in cooperation with the Research and Development Department of the former Immuno AG Company in Vienna, the infected cell extract in question was purified on lentil lectin to obtain a glycoprotein mix containing at least four envelope glycoproteins (gB, gC, gD and gG). This subunit vaccine was immunogenic and protective in mice as well as rabbits and showed at least partial cross protection in the HSV-2-challenged guinea pigs infected by the vaginal route.
\n
\n
\n
5.2. Recombinant HSV-1 and HSV-2 vaccines
\n
The next step of HSV vaccine development proceeded from purified (nearly cell DNA-free) subunit vaccines to those composed of recombinant HSV-1/HSV-2 polypeptides [86–90]. Great majority of the recombinant vaccines contained the gB and/or gD polypeptides, and their efficacy was done in mice. The first purified glycoprotein vaccines (gB1 and gD1) were prepared on immunoaffinity columns (with specific-bound MoAbs); such vaccines were found to protect mice against intracerebral challenge with HSV-1 [91–93]. Manservigi et al. [94] expressed the gB ectodomain in mammalian cells. After immunization with the purified gB fragment, the mice developed virus-neutralizing antibodies not only against HSV-1 but also cross-reacting antibodies to HSV-2. In addition, the authors also showed that the animals were resistant to the challenge with a lethal HSV dose. Later on, the recombinant proteins were prepared by transfection of plasmids (carrying the gD and/or gB ORFs) into competent Escherichia coli cells, which expressed the corresponding not glycosylated polypeptides [90, 95–100]. Recombinant glycoproteins (including gC) were also prepared in insect and/or mammalian cells, in which the recombinant products could be glycosylated [101].
\n
Immunization with the recombinant gD1 or gD2 alone, or in combination with the gB protected mice against infectious virus challenge given by intraperitoneal or subcutaneous routes and in the case of gD1 vaccination cross protection to HSV-2 was observed as well. Immunization with gD alone was more effective than that with gB alone [87], while immunity provided by gC alone was negligible [95]. In contrast, very high effect was achieved following immunization with the mix of gB1, gD1, and gE1 glycoproteins in combination with the Al(OH)3 adjuvant including the cross protection of guinea pigs challenged with HSV-2 at genital route [99]. A trivalent vaccine containing gD2, gC2 and gE2 prepared in the baculovirus expression system was more immunogenic for mice than the subunit vaccine containing gC2 and gD2 [102]. Furthermore, Awasthi et al. [103] immunized the guinea pigs with recombinant proteins gC2 and gD2 prepared in a baculovirus system (which activated antibody production), with the adenovirus vector carrying the VP5 capsid protein and with the tegument proteins VP13 and VP14 (which activated the T-cell-mediated response). However, no significant difference was detected, which confirmed that the glycoprotein mix gD2/gB2 was sufficient enough.
\n
The alum adjuvant in combination with the recombinant gD polypeptide activates the Th2-type humoral response and therefore may be less suitable for immunotherapeutic use, for which the Th1-type response is essential. To stimulate the Th1 response predominantly, cytokines as adjuvants were tested, such as IL-2 with good results after corneal challenge with the virulent HSV-1 CHR3 strain [104]. The so-called immune-stimulating lipid complexes (ISCOM) combined with HSV-2 immunogenic polypeptides induced both Th1- and Th2-type responses when showing a highly neutralizing antibody response along with the production of IL-2, IFN-gamma, and TNF [105]. Similar strong potentiating effect was observed at immunization with gD2 polypeptide adjuvanted with modified lipid A (AS04) [98]. Furthermore, when using the gD2 fusion polypeptide along with MF59 (squalene) as adjuvant, the local mucosa immune response could neither be stimulated properly, nor the latency reactivation prevented [106]. There is worth mentioning that the so-called autoimmune/inflammatory syndrome induced by adjuvants (ASIA) is an inflammatory syndrome associated with certain adjuvants (such as MF59/squalene); the latter potentiates pathological autoimmune reactions, which may be evident in the case of prophylactic mass immunization campaigns [107].
\n
In contrast to T-lymphocyte stimulation, the local IgA secretion can be enhanced by mistletoe lectins such as ML-1, ML-2, and/or ML-3, the application of which in combination with the gD2 polypeptide elicited a good secretory IgA response at intranasal challenge in mice [108]. The authors also used non-ionized liposome like particles carrying the gB ectodomain and additional polylysine-rich polypeptides. Intranasal application of the product in question provided protection against lethal challenge with HSV-2 and induced a strong Th1-type immune response [109]. Skoberne et al. [110] prepared an experimental vaccine named GEN-003/MM-2, which contained gD2 and the IE protein ICP4/175K expressed in a baculovirus system. In combination with MM2 adjuvants (also called matrix M2 composed of cholesterol, phospholipids, and saponin), the vaccine showed immunotherapeutic effect in guinea pigs when it reduced the frequency of recurrent genital lesions upon HSV-2 challenge.
\n
\n
\n
5.3. Viral vectored and DNA vaccines
\n
The recent idea of DNA vaccines represents a progressive approach for immunization (reviewed at [111]). The advantage of DNA vaccines is that the immune response resembles to that following the administration of live-attenuated virus vaccines, but without the risk of reversion to viral phenotype. The viral DNA vaccines fall into two categories: viral DNA carriers and the recombinant plasmid vaccines (classical DNA vaccines). The viral DNA carriers are in fact nonpathogenic attenuated viruses that genome contains the inserted ORF fragments encoding the desired antigen. The best examples are either the recombinant adenovirus or vaccinia virus, which carry the HSV gD or gB genes. The latter glycoproteins become expressed in the immunized animal, that is, in the mice, which are then protected against HSV challenge [112–115]. The recombinant vesicular stomatitis virus (VSV) was prepared from a plasmid containing the cDNA of the VSV RNA genome and the gD2 ORF US6. This recombinant virus induced a good cell-mediated immune response as well as anti-gD antibody formation. Immunized mice were protected against acute HSV-2 challenge, and the establishment of latency was also reduced [116]. Chiuppesi et al. [117] described the effect of immunization with feline lentiviral vector-based, herpes simplex virus 1 (HSV-1) glycoprotein B vaccine. This lentivirus construct induced HSV-1 antibody formation and also provided cross protection against lethal HSV-2 infection.
\n
Many classical plasmid DNA vaccines were prepared with the inserted HSV DNA fragment. The majority of them encoded HSV-1 and/or HSV-2 glycoproteins such as gB, gD, gC, and gE; their efficacy was tested in mice, guinea pigs, and rabbits [90, 118–124]. The results were obtained after immunization with plasmids encoding the glycoproteins gD and/or gB of either HSV-1 or HSV-2 alone or in combination. Especially the immunization of mice and/or guinea pigs with the gD-expressing plasmid protected against challenge with a lethal dose of HSV-2 [124–128], but without clear-cut reduction of the latency rate. Better results were obtained with plasmids encoding both gB and gD [119, 129, 130]. The efficiency of DNA vaccines could be higher when adding cytokines such as IL-12 which induced the Th type 1 immune response [131, 132] similarly as the presence of IL-18 and/or RANTES [133, 134]. In contrast, the Th2-type response could be better achieved if the gD and IL-4-coding plasmid was used for immunization [133, 135]. The Th type 2 immune response is important for virus clearance from peripheral tissues including the infection of the eye, while the Th1-type response was involved in preventing the latency establishment [135]. The DNA vaccine-encoding gB when combined with DNA plasmid coding for cytokines IL-12 and IL-18 as adjuvant was efficient in prevention of the vaginal infection in mice: gB1/IL-12 and/or gB1/IL-18 elicited a local resistance of genital mucosa and protected mice against lethal HSV-2 challenge. The best results were observed with IL-12, while additional adjuvants did not enhance protection [136]. Another adjuvant tested with the DNA vaccines was the lipid adjuvant Vaxfectin®. In guinea pigs immunized with plasmid-expressing gD2 and VP11/VP12 as well as VP13/14 polypeptides along with a lipid adjuvant, a great prophylactic effect and the reduced HSV-2 replication in the genital tract of experimental animals was observed. The vaccine also showed immunotherapeutic properties when reducing the extent of latency (vDNA contents) in the dorsal root ganglia [137, 138]. Immunization with the recombinant plasmid encoding the fusion protein consisting of the gB and the CCL19 chemokine ORF induced both the Th type 1 and the Th type 2 responses including an increased local secretory IgA antibody production protecting mice against HSV-2 challenge [139]. The enhanced efficiency of DNA vaccines could be achieved by combining genes encoding the envelope glycoproteins with certain nonstructural HSV genes. The recombinant plasmid consisting of gD2, gB2, and ICP27 ORFs elicited a higher cellular as well as humoral response than the plasmid construct encoding the glycoproteins only. The construct in question also provided a higher protection against vaginal challenge with HSV-2 [140].
\n
\n
\n
5.4. Live-attenuated HSV-1 and HSV-2 vaccines
\n
Attenuated HSV vaccines are live HSV-1 and/or HSV-2 viruses derived from the wild-type strains by modifications of their virulence but keep the immunogenic properties. The best known deletion for HSV-1 was the removal of the UL22 ORF-encoding gH glycoprotein [141, 142] and/or the removal of the IR1 sequence encoding the protein γ34.5 gamma, a neurovirulence factor [143]. Furthermore, the deletion of the gE encoding by US8 gene, which is not needed for virus replication in cell culture but is inevitable for neural uptake [144, 145] along with the deletion of the UL41 gene, encoding the virion host shutoff (vhs) protein [146] might be an excellent solution. In this respect, the non-virulent HSZP strain with natural mutations altering the vhs protein, if deleted in the gE gene, would be of special advantage [147]. Another attempts to prepare a live-attenuated HSV strain were made by deleting nonstructural genes, such as the ORF UL54 (encodes the ICP27 IE polypeptide), the ORF RS1 (encodes the transactivation protein ICP4) [146, 148], and/or the ORF RL2 (encoding the ICP0 transactivation protein) as described by Halford et al. [149]. All the above-mentioned attenuated viruses was protective against HSV challenge in various animal models. The HSV-2 deleted in the gH gene designated as disabled infectious single cycle (DISC) and tested as therapeutic vaccine, but no convincing protection was found [150].
\n
Several other attenuated HSV strains were prepared by deleting the nonstructural genes important for virulence. Deleted ORFs for such purpose were the following: (1) the UL23 ORF encoding the thymidine kinase (TK) as described by Morrison and Knipe [151], (2) the UL39 ORF encoding the large RR1 subunit [152], (3) the UL29 ORF encoding the ssDNA-binding ICP8 polypeptide [153], (4) the UL5 ORF encoding a protein of the primase/helicase complex [154, 155], and finally (5) the UL9 ORF encoding the ori-binding protein [156]. The TK minus recombinant R7017 was prepared from the w.t. strain F, which genome was, in addition, deleted in the IR1 ORF sequence encoding the γ34.5 and furthermore manipulated by inserting the gD2, gI2, and gG2 sequences along with a portion of the gE ORF (US4–US8). The TK-reversed recombinant virus was further modified by reinserting the TK gene (R7020 TK plus). Both recombinant viruses protected mice as well as guinea pigs against HSV-2 challenge [148]. The next UL39-/RR1-deleted virus was tested not only in animals but also in man [152, 157]. The phase I and phase II clinical trials showed partial protection against recurrences (in 37.5% of immunized individuals as compared with the placebo group) [157]. The attenuated strain deleted in the genes UL29 and UL5 (called dl29-5) protected mice against ocular infection with HSV-1 and guinea pigs against genital challenge with HSV-2 [158, 159]. Stanfield et al. [160] prepared an attenuated strain of HSV-2 called VC2, which had mutations in the gK gene and in the UL20 ORF (which is membrane protein inhibiting the neural uptake). This vaccine prevented genital infection upon vaginal challenge with HSV-2 and also inhibited latent infection in the lumbosacral dorsal root ganglia. Finally, the defective HSV-2 designated CJ9-gD2 was used to immunize mice. This virus had mutation in the UL9 ORF which encodes a vDNA replication protein and was deleted in the gD2 ORF (US9). The attenuated vaccine induced a higher antibody response in comparison with the gD2-alum/MPL subunit vaccine (used in a human trial) and protected mice against lethal challenge with HSV-2 [161].
\n
\n
\n
\n
6. Human vaccination trials: prophylactic and therapeutic HSV vaccines
\n
Some of the vaccines mentioned above were tested not only in animal models but also in man at clinical trials. They were designed either as prophylactic vaccines, or they were destined for immunotherapeutic use, having been suggested for prevention or modulation of genital infection predominantly caused by HSV-2. The aim of prophylactic vaccine against HSV-2 infection is not only to prevent a clinical disease at primoinfection but also to interfere with the subsequent establishment of latency. The latter is a hard task, since the majority of vaccines just reduce the number of neurons which are getting HSV DNA carriers but does not fully prevent latent infection. Therefore, it is more reasonable to design a vaccine for immunotherapeutic use aimed to reduce the extent and the frequency of recurrent lesions.
\n
The prophylactic vaccine should induce a satisfactory humoral as well as cell-mediated immune response. For such purpose, subunit, recombinant and DNA vaccines were tested. The first subunit vaccine tested was an HSV-2 glycoprotein mix (gB, gC, gG, gD, and gE) purified from infected chick embryo cells (this product is essentially similar to the subunit HSZP vaccine immuno mentioned above). As shown in 22 seronegative volunteers, this particular vaccine induced both the humoral and the cell-mediated specific response [162]. Alternatively, a similar vaccine (containing gB2 and gG2) was used to immunize 161 seronegative individuals with a less encouraging effect at phase II clinical trial [163]. The company Chiron Corporation sponsored the testing of a recombinant gB2/gG2 vaccine mixed with the MF59 adjuvant used for immunization of 137 persons. The efficiency of this vaccine was reported 9% only [53, 164]. Another study by GlaxoSmithKline uses a recombinant gD2 in combination with alum adjuvant and/or ASO4; in the latter trial, 7460 volunteers were selected as seronegative before the onset of the phase II trial. Immunization with the vaccine in question induced higher antibody titers than natural infection and also conferred partial protection against natural infection that had a milder course as compared to non-immunized controls [165]. At phase III trial, 847 seronegative individuals (no antibodies to HSV-1 as well as to HSV-2) and 1867 women showing antibodies to HSV-1 but not to HSV-2 were immunized with the same vaccine [166]. The results showed that 73–74% immunized women which were seronegative against both antigens have been protected against HSV-2 primoinfection (as detected by natural seroconversion), but the vaccine was not effective in man and in HSV-1 seropositive women. The GlaxoSmithKline company provided another trial, during which together 8323 seronegative women (for HSV-1 as well as HSV-2) were immunized. This follow-up was less encouraging when showing only 20% efficiency when measured by the seroconversion rate against natural genital HSV-2 infection and about 58% protection against HSV-1 genital infection [167]. In a multicenter study from 2013, the possible role of the abovementioned gD2-ASO4 vaccine in the etiology of stillbirth was evaluated. Together 19,727 pregnant women were immunized, from which 13.3% really had stillbirth in comparison with 11.00% of pregnant women in the control group that shows a clearly not significant effect [168].
\n
The HSV DNA vaccine was also tested for prophylactic use. In the phase I trial, 62 seronegative women were immunized with the gD2 recombinant plasmid GENEVAX. T-cell-mediated response was found in one out of four volunteers [169]. Another prophylactic candidate vaccine tested was the HSV-2/HSV529-attenuated virus (deleted in genes UL5 and and UL29, also called dl5-29). This vaccine when tested in guinea pigs elicited a satisfactory humoral as well as T-cell response, conferred protection to HSV-2 challenge, and reduced the extent of latency in the regional lumbosacral ganglia [170]. The vaccine efficiency in human will be evaluated by the end of 2016.
\n
As already mentioned, the therapeutic vaccines should decrease the frequency of recurrence episodes as well as their severity. To achieve this, the therapeutic vaccine must induce a potent and specific T-cell-mediated response. Several subunit vaccines were tested with an immunotherapeutic purpose, namely, the semipurified inactivated infected cell extracts, recombinant vaccines, and attenuated viruses. Probably, the first of the purified inactivated HSV-1-infected cell extract tested was the vaccine by Kutinová et al. [171]. A similar vaccine introduced by Skinner et al. [172] was used for immunization of 316 subjects with herpes disease who had recurrent blisters at least six times per year. Though this vaccine induced a detectable increase in the antibody levels and a satisfactory T-cell-mediated response, no difference in the frequency of recurrences was noted during the 1-year observation period. However, the vaccination accelerated the healing of the lesions, which had a less severe course in the immunized persons. Among the recombinant vaccines tested in a therapeutic context, the gD2 vaccine containing alum adjuvant should be mentioned first. This was used for immunization of 98 HSV-2 seropositive persons who had at least four but up to 14 recurrence episodes per year [173]. In this trial, the average of six recurrences per year in the placebo group was siginificantly higher than the average of four recurrences per year in the immunized group (p = 0.039). Furthermore, the gB2/gD2 vaccine mixed with the MF59 adjuvant was applied in another trial, in which 202 volunteers were immunized. At the phase II clinical trial, no difference was found in the titers of neutralizing antibodies during the observation period of 8 months, but there was a significant difference in the increased interval between the occurrences of the recurrent lesions following the immunization procedure [174]. Further trials were made using peptide vaccines. An overview of defined T-cell-based epitopes from HSV proteins was reported by Laing et al. ([175], Table 2).
\n
Not all known T-cell based epitopes can induce protective immunity. Mapping of T cells in seropositive population found out that CD4+ T cells were mainly stimulated by tegument proteins UL21, UL46, UL47, UL49 and envelope glycoproteins gB and gD. It was observed that gB stimulates both CD4+ and CD8+ T cells, whereas gD induces stronger CD4+ T-cell-dependent immunity [58]. CD8+ T cells were stimulated by gD53–61, gD70–78, and gD278–286 peptides mainly in HLA-A*02:01-positive HSV-1 and HSV-2 seropositive healthy individuals [190]. Moreover, CD8+ T cells were also induced by tegument protein VP11/VP12 (gene UL46) that would account for another candidate protein to prepare effective HSV vaccine [61].
Vaccine design also requires involvement of epitopes suitable for HLA binding by most people. In most ethnic groups, HLA-A*02:01 and HLA-B*07:02 belong to the most abundant HLA alleles [194]. HLA-A*02:01-restricted epitopes have been found in glycoprotein B (UL27 442–451), tegument protein VP13/VP14 (UL47 551–559), and tegument protein coded for UL25 (UL25 372–380) of HSV-2, and they stimulated CD8+ T-cell response [176, 195]. HLA-B*07:02-restricted epitopes have been reported for tegument protein VP22 (gene UL49) (HSV-2 UL49 49–57, HSV-2 UL49 82–90, HSV-2 UL49 99–108, HSV-2 UL49 131–140, and HSV-1 UL49 291–290) and stimulated CD8+ T-cell response [176]. Samandary et al. [196] found association with high prevalence of herpes infection and disease with the frequency of HLA-A*24, HLA-B*27, and HLA-B*58 alleles. In contrast, low prevalence of herpes infection and disease appeared associated with the high frequency of HLA-B*44 allele.
\n
The effectivity of immune response is also depending on adjuvant type. Cooper et al. [189] found out that vaccine design and adjuvant type can have a significant effect on T-cell epitope utilization. Four epitopes within the gD2 molecule gD2 49–63, gD2 105–119, gD2 245–259 and gD2 333–347 were administered to mice with alum or IL-12. CD4+ T-cell response was induced in mice immunized with gD HSV-2 epitope gD2 245–259 and adjuvant alum. Mice immunized with IL-12 stimulate CD4+ T-cell response to HSV-2 epitope gD2 245–259 as well as to gD2 333–347 [189].
\n
“Epitope” (peptide) vaccines that selectively stimulate T and B cells belong to other herpes simplex vaccine candidates. Wang et al. [180] prepared multi-epitope peptide vaccine that contained six B-cell epitopes from different glycoproteins of HSV-2 (gB2 466–473, gC2 216–223, gD2 6–18, gE2 483–491, gG2 572–579 and gI2 286–295), four CD4+ T-cell-based epitopes (gD2 21–28, gD2 205–224, gD2 245–259 and gB2 162–177) and two CD8+ T-cell-based epitopes (gD2 10–20 and gD2 268–276). All above-described epitopes were inserted into the extracellular fragment (1–290) of HSV-2 glycoprotein D to construct multi-epitope assembly peptides (MEAPs) by replacing some non-epitope amino acid sequences. The genes of the selected peptides were inserted into recombinant plasmid and expressed in E. coli strain BL21. The multi-epitope vaccine elicited in mice production of virus-neutralizing antibodies induced Th1 and Th2 immune response and protected mice against intravaginally induced lethal challenge of HSV-2 [180].
\n
Many studies have focused on mapping of protective epitopes that stimulate immunity in asymptomatic individuals, that is, individuals without clinical findings of herpes infection. Analysis of IFN-γ-producing CD4+ T cells in HSV-1 seropositive individuals revealed that gB peptide epitopes (aa 166–180 and aa 666–680) were strongly recognized by CD4+ T cells from asymptomatic individuals, but not from symptomatic individuals. Inversely, CD4+ T cells from symptomatic individuals preferentially recognized gB (aa 661–675) [179]. Another study identified asymptomatic CD8+ T-cell epitopes from glycoprotein D (gD53–61, gD70–78 and gD278–286) [191]. It can be hypothesized that repertoire of T-cell-based epitopes determines either the development of HSV-1 (-2) clinical symptoms or asymptomatic viral shedding.
\n
Finally, some investigators preferred the genetically attenuated virus vaccines, because they induce an immune response essentially similar to that following natural infection. For example, Casanova et al. [157] immunized the volunteers (32 persons having at least five recurrent lesions per year) with an attenuated HSV-2 virus deleted in the UL39 gene (the RR1 protein ORF). The phase I and phase II clinical trials showed the reduction of recurrent clinical manifestations by 37.5% as compared with the placebo-immunized group. Another clinical trial, which was performed in 2006, took the advantage of HSV-2-attenuated virus deleted in the gH gene (the so-called disabled infectious single cycle [DISC] virus). Unfortunately, no significant difference was found in the number of recurrences between the immunized and control mock-immunized subjects during the 1 year follow-up period [150]. The last phase I and II clinical trial which results will be announced nowadays concerns volunteers who had from two to nine recurrent lesions per year; they were immunized with a DNA vaccine-expressing gD2 in combination with the lipid adjuvant Vaxfectin [197].
\n
\n
\n
7. Future perspectives of HSV-1/HSV-2 vaccination with the emphasis on the T-cell response
\n
As described above, an effective vaccine against HSV-1 and HSV-2 infection that would prevent virus reactivation (therapeutic vaccine) should stimulate both humoral and cellular immunity mediated by CD4+ and CD8+ T cells. This immune response can be induced by live-attenuated virus, but such viruses are not safe because of their possible reversion back to the wild-type virus. Therefore, alternative approaches to develop effective herpes simplex vaccine have been attempted during the last decades. Nowadays, T-cell-inducing herpes simplex vaccines bearing “asymptomatic” immunodominant epitopes derived from HSV proteins were designed and tested [180]. Such vaccines possess many advantages over traditional vaccine like (1) induction of specific T-cell-based immunity, (2) inhibition of pathogenic immune response, and (3) safety.
\n
Only few asymptomatic T-cell epitope-based vaccines were prepared and tested until now. The vaccine used by Wald et al. [198] contained 32 synthetic immunogenic HSV-2 peptides, linked to the heat shock protein Hsp 70 in combination with the QS-21 (contains a saponin) adjuvant. In 32 immunized volunteers, the vaccine elicited no complication, but induced a satisfactory CD4+/CD8+ T-lymphocyte response against a wide range of immunogenic HSV-2 peptides used in the stimulation tests in vitro. Chentoufi et al. [191] prepared CD8+ T-cell epitope-based vaccine containing three separate pairs of CD4–CD8 lipopeptides. Each of the lipopeptide contained one of the three asymptomatic immunodominant human CD8+ T-cell peptide epitopes from HSV-1 glycoprotein D (gD53–61, gD70–78, and gD278–286) that were joined with a human CD4+ T-cell peptide epitope (gD49–82). Humanized HLA-A*02:01 transgenic rabbits were immunized with a mixture of the three CD4–CD8 HSV-1 gD lipopeptides. Immunization induced an increased production of CD4+ and CD8+ T cells and protected rabbits against ocular HSV disease [191]. The same ASYMP vaccine was used for therapeutic vaccination of HLA transgenic rabbits infected by HSV-1. The vaccine induced production of HSV-specific CD8+ T cells that prevent HSV-1 reactivation ex vivo from latently infected explanted trigeminal ganglia. Moreover, the vaccine significantly reduced HSV-1 shedding and boosted the function of HSV-1 gD epitope-specific CD8+ T cells in draining lymph nodes, conjunctiva, and trigeminal ganglion [199].
\n
The lipopeptide vaccines belong to another herpes simplex vaccine candidate. The lipopeptide vaccine can be easily produced and possesses some advantages over traditional vaccine such as safety and tolerance, bearing protective T-cell epitopes derived from HSV antigens, missing of non-immunogenic harmful epitopes, and missing of viral pathogenic proteins such as ICP47, and lipids have functioned as adjuvant. Previous studies observed that lipopeptide vaccine injected intranasally into mice induced mucosal and systemic B and Th1 immune response [191]. Other authors prefer attenuated genetically modified herpes simplex viruses as successfully vaccine candidates (Section 5.4).
\n
\n
\n
8. Conclusions
\n
In conclusion, the development of successfully therapeutical vaccine against HSV infection should respect the following recommendations: (1) the assessment of putative differences in the recognition of T- and/or B-cell epitopes from envelope, tegument, and regulatory HSV proteins in patients with recurrent herpes disease versus asymptomatic individuals; (2) the production of new safer adjuvants avoiding those claimed to cause the ASIA syndrome; (3) the induction of local mucosal immunity mediated by lipopeptides; (4) the use of humanized susceptible HLA transgenic mice as well as rabbits before human trials; and (5) at last but not least, the efficacy of human trials for an immunotherapeutic vaccine that should be made according to internationally accepted unified criteria in at least three groups of subjects (seronegative individuals, seropositive individuals without recurrent herpes disease, and volunteers with such disease). The tests should include the demonstration of elevated antibody titers after vaccination and in vitro testing of the T-lymphocyte response in a blastic transformation test as well as production of selected cytokines; 6. Finally, the application of the therapeutic vaccine in future human trials should be intracutaneous rather than by using other administration routes.
\n
\n\n',keywords:"herpes simplex virus 1 and herpes simplex virus 2, candidate vaccine, clinical trials, efficacy in animals, humoral and cell-mediated immune response, antigenic epitopes",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/51617.pdf",chapterXML:"https://mts.intechopen.com/source/xml/51617.xml",downloadPdfUrl:"/chapter/pdf-download/51617",previewPdfUrl:"/chapter/pdf-preview/51617",totalDownloads:917,totalViews:313,totalCrossrefCites:2,totalDimensionsCites:2,hasAltmetrics:0,dateSubmitted:"March 9th 2016",dateReviewed:"May 31st 2016",datePrePublished:null,datePublished:"September 7th 2016",readingETA:"0",abstract:"This chapter is devoted to the topics of not yet marketed HSV vaccine, which is still in the focus of interest, especially from the point of immunotherapeutic use. To understand the principles of vaccination strategies (prophylactic and/or immunotherapeutic), the pathogenesis of herpes simplex virus 1 (HSV-1) and/or HSV-2 infections in animal models is briefly outlined. Even when both herpesviruses may spread via bloodstream, which is especially true in the immunocompromised host, the main route of their transmission is along peripheral nerves. Both viruses establish latency in ganglion cells, and after reactivation, they spread along axons back to the site of primary infection. Since neither the establishment of latency nor its reactivation can be fully controlled by virus-neutralizing antibodies, the outcome of immune response greatly depends on the activity of cytotoxic CD8+ T lymphocytes. The majority of important antigenic epitopes is located in envelope glycoproteins (such as gB, gD, gE, gC and gG) that are related to virus adsorption and penetration into susceptible cells. The HSV-1 and/or HSV-2 experimental vaccines designed so far were either purified virion products derived from infected cells (subunit vaccines), purified recombinant immunogenic herpes simplex virus HSV-coded proteins (especially gD), and/or attenuated live viruses lacking some of virulence tools (such as gH and/or gE). We bring a comprehensive overview of the efficacy of experimental HSV-1/HSV-2 vaccines and discuss our own data. In conclusion, we believe in the continued demand of HSV-1 and HSV-2 vaccines, at least for their immunotherapeutic use, suggesting unified evaluation criteria for clinical trials to reach consent at their interpretation.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/51617",risUrl:"/chapter/ris/51617",book:{slug:"herpesviridae"},signatures:"Vladimíra Ďurmanová, Marian Adamkov and Július Rajčáni",authors:[{id:"181773",title:"Emeritus Prof.",name:"Julius",middleName:null,surname:"Rajčáni",fullName:"Julius Rajčáni",slug:"julius-rajcani",email:"viruraj@savba.sk",position:null,institution:{name:"Slovak Academy of Sciences",institutionURL:null,country:{name:"Slovakia"}}},{id:"185924",title:"Prof.",name:"Marian",middleName:null,surname:"Adamkov",fullName:"Marian Adamkov",slug:"marian-adamkov",email:"marian.adamkov@jfmed.uniba.sk",position:null,institution:null},{id:"186238",title:"Associate Prof.",name:"Vladimira",middleName:null,surname:"Ďurmanová",fullName:"Vladimira Ďurmanová",slug:"vladimira-durmanova",email:"vladimira.durmanova@fmed.uniba.sk",position:null,institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. The molecular biology of HSV: virion structure, lytic replication and latency",level:"1"},{id:"sec_3",title:"3. Pathogenesis of HSV-1 and/or HSV-2 infections",level:"1"},{id:"sec_4",title:"4. Mechanisms of the immune response to HSV-1 or HSV-2",level:"1"},{id:"sec_4_2",title:"4.1. The innate immune response",level:"2"},{id:"sec_5_2",title:"4.2. The acquired immune response",level:"2"},{id:"sec_7",title:"5. Survey of experimental HSV-1 and HSV-2 vaccines and their design",level:"1"},{id:"sec_7_2",title:"5.1. Inactivated virion and subunit vaccines",level:"2"},{id:"sec_8_2",title:"5.2. Recombinant HSV-1 and HSV-2 vaccines",level:"2"},{id:"sec_9_2",title:"5.3. Viral vectored and DNA vaccines",level:"2"},{id:"sec_10_2",title:"5.4. Live-attenuated HSV-1 and HSV-2 vaccines",level:"2"},{id:"sec_12",title:"6. Human vaccination trials: prophylactic and therapeutic HSV vaccines",level:"1"},{id:"sec_13",title:"7. Future perspectives of HSV-1/HSV-2 vaccination with the emphasis on the T-cell response",level:"1"},{id:"sec_14",title:"8. Conclusions",level:"1"}],chapterReferences:[{id:"B1",body:'\nGruter W. Das Herpesvirus, seine aetiologische und klinische Bedeutung. Munch Med Wsch 1924, 71: 1058.\n'},{id:"B2",body:'\nSchneweis KE. Untersuchungen zur Typendifferenzierung des Herpesvirus hominis. Z Immun-Forsch 1962, 124: 24–28.\n'},{id:"B3",body:'\nWildy P. Classification and nomenclature of viruses, pp. 33–34, In: Monographs in Virology, Melnick JL (ed), Karger, Basel, 1971.\n'},{id:"B4",body:'\nLee EK, Coleman RM, Pereira L, Bailey PD, Tatsuno M, Nahmias AJ. Detection of herpes simplex virus type 2 specific antibody with glycoprotein G. J Clin Microbiol 1985, 22: 641–644.\n'},{id:"B5",body:'\nLiu F, Zhou ZH. Comparative virion structures of human herpesviruses, pp. 27–43, In: Arvin A, Campadelli-Fiume G, Mocarski E, Moore PS, Roizman B, Whitley R and Yamanishi K (eds). Human Herpesviruses: Biology, Therapy and Immunoprophylaxis. Cambridge University Press, 2007.\n'},{id:"B6",body:'\nRoizman B, Knipe DM, Whitley RJ. Herpes Simplex Viruses, pp. 1823–1897, In: Knipe DM, Howley PM (eds) Fields Virology, 6th ed., Walters Cluver/Lippincott Williams and Wilkins, Philadelphia, 2013.\n'},{id:"B7",body:'\nKnipe DM, Cliffe AR. Chromatin control of herpes simplex virus lytic and latent infection. Nat Rev Microbiol 2008, 6: 211–221. doi: 10.1038/nrmicro1794.\n'},{id:"B8",body:'\nCampadelli-Fiume G, Menotti L, Avitabile E, Gianni T. Viral and cellular contributions to herpes simplex virus entry into the cell. Curr Opin Virol 2012, 2: 28–36. doi: 10.1016/j.coviro.2011.12.001.\n'},{id:"B9",body:'\nRajčáni J, Vojvodová A. The role of herpes simplex virus glycoproteins in the virus replication cycle. Acta Virol 1998, 42: 103–118.\n'},{id:"B10",body:'\nGianni T, Amasio M, Campadelli-Fiume G. Herpes simplex virus gD forms distinct complexes with fusion executors gB and gH/gL in part through the C-terminal profusion domain. J Biol Chem 2009, 284: 17370–17382. doi: 10.1074/jbc.M109.005728.\n'},{id:"B11",body:'\nSatoh T, Arii J, Suenaga T, Wang J, Kogure A, Uehori J, Arase N, Shiratori I, Tanaka S, Kawaguchi Y, Spear PG, Lanier LL, Arase H. PILRα is a herpes simplex virus-1 entry co-receptor that associates with glycoprotein B. Cell 2008, 132: 935–944. doi: 10.1016/j.cell.2008.01.043.\n'},{id:"B12",body:'\nStampfer SD, Lou H, Cohen GH, Eisenberg RJ, Heldwein EE. Structural basis of local, pH dependent conformational changes in glycoprotein B from herpes simplex virus type 1. J Virol 2010, 84: 12.924–12.933. doi: 10.1128/JVI.01750-10.\n'},{id:"B13",body:'\nDingwell KS, Brunetti CR, Hendricks RL, Tang Q, Tang M, Rainbow AJ, Johnson DC. Herpes simplex virus glycoprotein E and I facilitate cell to cell spread in vivo and across junctions of cultured cells. J Virol 1994, 68: 834–845.\n'},{id:"B14",body:'\nRajčáni J, Ďurmanová V. Early expression of herpes simplex virus (HSV) proteins and reactivation of latent infection. Folia Microbiol 2000, 45: 7–28\n'},{id:"B15",body:'\nRežuchová I, Kúdelová M, Ďurmanová V, Vojvodová A, Košovský J, Rajčáni J. Transcription at early stages of herpes simplex virus 1 infection and during reactivation. Intervirology 2003, 46: 25–34.\n'},{id:"B16",body:'\nStevens JG, Wagner EK, Devi-Rao GB, Cook ML, Feldman LT. RNA complementary to a herpesvirus alpha gene mRNA is prominent in latently infected neurons. Science 1987, 235: 1056–1059.\n'},{id:"B17",body:'\nAhmed MM, Lock C, Millerm G. Regions of the herpes simplex virus type 1 latency-associated transcript that protect cells from apoptosis in vitro and protect neuronal cells in vivo. J Virol 2002, 76: 717–722.\n'},{id:"B18",body:'\nPerng GC, Jones C, Ciacci-Zanella J, Stone M, Henderson G, Yukht A, Slanina SM, Hofman FM, Ghiasi H, Nesburn AB, Wechsler SL. Virus induced neuronal apoptosis blocked by the herpes simplex virus latency associated transcript (LAT). Science 2000, 287: 1500–1503.\n'},{id:"B19",body:'\nPerng GC, Maguen B, Jin L, Mott KR, Osorio N, Slanina SM, Yukht A, Ghiasi H, Nesburn AB, Inman M, Henderson G, Jones C, Wechsler SL. A gene capable of blocking apoptosis can substitute for herpes simplex virus type 1 latency associated transcript gene and restore wild type reactivation levels. J Virol 2002, 76: 1224–1235\n'},{id:"B20",body:'\nEfstathiou S, Minson AC, Field HJ, Anderson JR, Wildy P. Detection of herpes simplex virus specific DNA sequences in latently infected mice and in humans. J Virol 1986, 32: 446–455.\n'},{id:"B21",body:'\nKristie TM and Roizman B. Differentiation and DNA contact points to the host proteins binding at the cis site of for virion-mediated induction of herpes simplex virus 1 alpha genes. J Virol 1988, 62: 1145–1157\n'},{id:"B22",body:'\nLillycrop KA, Budrahan VS, Lakin ND, Terrenghi G, Wood JN, Polak JM, Latchman DS. A novel POU family transcription factor is closely related to Brn-3 but has a distinct expression pattern in neuronal cells. Nucleic Acids Res 1992, 20: 5093–5096.\n'},{id:"B23",body:'\nLillycrop KA, Estridge JK, Latchman DS. Functional interaction between different isoforms of the Oct-2 transcription factor expressed in neuronal cells. Biochem J 1994, 298: 245–248.\n'},{id:"B24",body:'\nPreston CM, Efstathiou S. Molecular basis of HSV latency and reactivation, pp. 602–615, In: Arvin A, Campadelli-Fiume G, Mocarski E, Moore PS, Roizman B, Whitley R, Yamanishi K. (eds.) Human Herpesviruses, Biology, Therapy and Immunoprophylaxis, Cambridge University Press, 2007.\n'},{id:"B25",body:'\nFries LF, Friedman HM, Cohen GH, Eisenberg RJ, Hammer CH, Frank MM. Glycoprotein C of herpes simplex virus 1 is an inhibitor of the complement cascade. J Immunol 1986, 137: 1636–1641.\n'},{id:"B26",body:'\nHill TJ, Field HJ, Blyth WA. Acute and recurrent infection with herpes simplex virus in the mouse: a model for studying latency and recurrent disease. J Gen Virol 1975, 28: 341–353.\n'},{id:"B27",body:'\nHill TJ, Blyth HJ. An alternative theory of herpes simplex recurrence and a possible role for prostaglandins. Lancet 1976, 1: 397–398.\n'},{id:"B28",body:'\nWalz M, Price R, Notkins A. Latent ganglionic infection with herpes simplex virus types 1 and 2: viral reactivation after neurectomy. Science 1974, 184: 1185–1187.\n'},{id:"B29",body:'\nKúdelová M, Rajčáni J. Herpes simplex virus and human CNS infection, pp. 169–214, In: Singh SK, Ružek D. (eds), Neuroviral Infection, 1st ed., Taylor and Francis Group CRC, 2013.\n'},{id:"B30",body:'\nWhitley R, Kimberlin D, Prober Ch. HSV 1 and 2: pathogenesis and disease. pp. 569–601, In: Arvin A, Campadelli-Fiume G, Mocarski E, Moore PS, Roizman B, Whitley R, Yamanishi K. (eds.). Human Herpesviruses, Biology, Therapy and Immunoprophylaxis, Cambridge University Press, 2007.\n'},{id:"B31",body:'\nSvensson A, Bellner L, Magnusson M, Eriksson K. Role of IFN-alpha/beta signaling in the prevention of genital herpes virus type 2 infection. J Reprod Immunol 2007, 74: 114–123.\n'},{id:"B32",body:'\nConrady CD, Halford WP, Carr DJ. Loss of the type I interferon pathway increases vulnerability of mice to genital herpes simplex virus 2 infection. J Virol 2011, 85: 1625–1633. doi: 10.1128/JVI.01715-10.\n'},{id:"B33",body:'\nKim M, Osborne NR, Zeng W, Donaghy H, McKinnon K, Jackson DC, Cunningham AL. Herpes simplex virus antigens directly activate NK cells via TLR2, thus facilitating their presentation to CD4 T lymphocytes. J Immunol 2012, 188: 4158–4170. doi: 10.4049/jimmunol.1103450.\n'},{id:"B34",body:'\nLundberg P, Welander P, Han X, Cantin E. Herpes simplex virus type 1 DNA is immunostimulatory in vitro and in vivo. J Virol 2003, 77: 11158–11169.\n'},{id:"B35",body:'\nGuo Y, Audry M, Ciancanelli M, Alsina L, Azevedo J, Herman M, Anguiano E, Sancho-Shimizu V, Lorenzo L, Pauwels E, Philippe PB, Pérez de Diego R, Cardon A, Vogt G, Picard C, Andrianirina ZZ, Rozenberg F, Lebon P, Plancoulaine S, Tardieu M, Valérie D, Jouanguy E, Chaussabel D, Geissmann F, Abel L, Casanova JL, Zhang SY. Herpes simplex virus encephalitis in a patient with complete TLR3 deficiency: TLR3 is otherwise redundant in protective immunity. J Exp Med 2011, 208: 2083–2098. doi: 10.1084/jem.20101568.\n'},{id:"B36",body:'\nHerman M, Ciancanelli M, Ou YH, Lorenzo L, Klaudel-Dreszler M, Pauwels E, Sancho-Shimizu V, Pérez de Diego R, Abhyankar A, Israelsson E, Guo Y, Cardon A, Rozenberg F, Lebon P, Tardieu M, Heropolitanska-Pliszka E, Chaussabel D, White MA, Abel L, Zhang SY, Casanova JL. Heterozygous TBK1 mutations impair TLR3 immunity and underlie herpes simplex encephalitis of childhood. J Exp Med 2012, 209: 1567–1582. doi: 10.1084/jem.20111316.\n'},{id:"B37",body:'\nAnk N, West H, Bartholdy C, Eriksson K, Thomsen AR, Paludan SR. Lambda interferon (IFN-lambda), a type III IFN, is induced by viruses and IFNs and displays potent antiviral activity against select virus infections in vivo. J Virol 2006, 80: 4501–4509.\n'},{id:"B38",body:'\nLi J, Hu S, Zhou L, Ye L, Wang X, Ho J, Ho W. Interferon lambda inhibits herpes simplex virus type I infection of human astrocytes and neurons. Glia 2011, 59: 58–67. doi: 10.1002/glia.21076.\n'},{id:"B39",body:'\nLopušná K, Režuchová I, Kabát P, Kúdelová M. Interferon lambda induces antiviral response to herpes simplex virus 1 infection. Acta Virol 2014, 58: 325–332.\n'},{id:"B40",body:'\nGrubor-Bauk B, Arthur JL, Mayrhofer G. Importance of NKT cells in resistance to herpes simplex virus, fate of virus-infected neurons, and level of latency in mice. J Virol 2008, 82: 11073–11083. doi: 10.1128/JVI.00205-08.\n'},{id:"B41",body:'\nAdler H, Beland JL, Del-Pan NC, Kobzik L, Sobel RA, Rimm IJ. In the absence of T cells, natural killer cells protect from mortality due to HSV-1 encephalitis. J Neuroimmunol 1999, 93: 208–213.\n'},{id:"B42",body:'\nMilligan GN. Neutrophils aid in protection of the vaginal mucosae of immune mice against challenge with herpes simplex virus type 2. J Virol 1999, 73: 6380–6386.\n'},{id:"B43",body:'\nKodukula P, Liu T, Rooijen NV, Jager MJ, Hendricks RL. Macrophage control of herpes simplex virus type 1 replication in the peripheral nervous system. J Virol 1999, 162: 2895–2905.\n'},{id:"B44",body:'\nBuc M. Interferon gamma (IFN-gamma), p. 95, In: Basic and Clinical Immunology, Comenius University, Bratislava, 2008.\n'},{id:"B45",body:'\nKassim SH, Rajasagi NK, Zhao X, Chervenak R, Jennings SR. In vivo ablation of CD11c-positive dendritic cells increases susceptibility to herpes simplex virus type 1 infection and diminishes NK and T-cell responses. J Virol 2006, 80: 3985–3993.\n'},{id:"B46",body:'\nHochrein H, Schlatter B, O\'Keeffe M, Wagner C, Schmitz F, Schiemann M, Bauer S, Suter M, Wagner H. Herpes simplex virus type-1 induces IFN-alpha production via Toll-like receptor 9-dependent and -independent pathways. Proc Natl Acad Sci U S A 2004, 101: 11416–11421.\n'},{id:"B47",body:'\nden Haan JM, Bevan MJ. Antigen presentation to CD8+ T cells: cross-priming in infectious diseases. Curr Opin Immunol 2001, 13: 437–441.\n'},{id:"B48",body:'\nRamirez MC, Sigal LJ. The multiple routes of MHC-I cross-presentation. Trends Microbiol 2004, 12: 204–207.\n'},{id:"B49",body:'\nLubinski J, Wang L, Mastellos D, Sahu A, Lambris JD, Friedman HM. In vivo role of complement-interacting domains of herpes simplex virus type 1 glycoprotein gC. J Exp Med 1999, 190: 1637–1646.\n'},{id:"B50",body:'\nMcDermott MR, Brais LJ, Evelegh MJ. Mucosal and systemic antiviral antibodies in mice inoculated intravaginally with herpes simplex virus type 2. J Gen Virol 1990, 71: 1497–1504.\n'},{id:"B51",body:'\nDudley KL, Bourne N, Milligan GN. Immune protection against HSV-2 in B-cell-deficient mice. Virology 2000, 270: 454–463.\n'},{id:"B52",body:'\nMorrison LA, Zhu L, Thebeau LG. Vaccine-induced serum immunoglobin contributes to protection from herpes simplex virus type 2 genital infection in the presence of immune T cells. J Virol 2001, 75: 1195–1204.\n'},{id:"B53",body:'\nCorey L, Langenberg AG, Ashley R, Sekulovich RE, Izu AE, Douglas JM Jr., Handsfield HH, Warren T, Marr L, Tyring S, DiCarlo R, Adimora AA, Leone P, Dekker CL, Burke RL, Leong WP, Straus SE. Recombinant glycoprotein vaccine for the prevention of genital HSV-2 infection: two randomized controlled trials. Chiron HSV Vaccine Study Group. JAMA 1999, 282: 331–340.\n'},{id:"B54",body:'\nHandunnetthi L, Ramagopalan SV, Ebers GC, Knight JC. Regulation of major histocompatibility complex class II gene expression, genetic variation and disease. Genes Immun 2010, 11: 99–112. doi: 10.1038/gene.2009.83.\n'},{id:"B55",body:'\nMilligan GN, Bernstein DI. Interferon-gamma enhances resolution of herpes simplex virus type 2 infection of the murine genital tract. Virology 1997, 229: 259–268.\n'},{id:"B56",body:'\nMilligan GN, Bernstein DI, Bourne N. T lymphocytes are required for protection of the vaginal mucosae and sensory ganglia of immune mice against reinfection with herpes simplex virus type 2. J Immunol 1998, 160: 6093–6100.\n'},{id:"B57",body:'\nGill N, Ashkar AA. Overexpression of interleukin-15 compromises CD4-dependent adaptive immune responses against herpes simplex virus 2. J Virol 2009, 83: 918–926. doi: 10.1128/JVI.01282-08.\n'},{id:"B58",body:'\nKoelle DM, Schomogyi M, McClurkan C, Reymond SN, Chen HB. CD4 T-cell responses to herpes simplex virus type 2 major capsid protein VP5: comparison with responses to tegument and envelope glycoproteins. J Virol 2000, 74: 11422–11425.\n'},{id:"B59",body:'\nZhu J, Koelle DM, Cao J, Vazquez J, Huang ML, Hladik F, Wald A, Corey L. Virus-specific CD8+ T cells accumulate near sensory nerve endings in genital skin during subclinical HSV-2 reactivation. J Exp Med 2007, 204: 595–603.\n'},{id:"B60",body:'\nKhanna KM, Bonneau RH, Kinchington PR, Hendricks RL. Herpes simplex virus-specific memory CD8+ T cells are selectively activated and retained in latently infected sensory ganglia. Immunity 2003, 18: 593–603.\n'},{id:"B61",body:'\nJing L, Haas J, Chong TM, Bruckner JJ, Dann GC, Dong L, Marshak JO, McClurkan CL, Yamamoto TN, Bailer SM, Laing KJ, Wald A, Verjans GM, Koelle DM. Cross-presentation and genome-wide screening reveal candidate T cells antigens for a herpes simplex virus type 1 vaccine. J Clin Invest 2012, 122: 654–763. doi: 10.1172/JCI60556.\n'},{id:"B62",body:'\nGebhardt BM, Hill JM. Cellular neuroimmunologic responses to ocular herpes simplex virus infection. J Neuroimmunol 1990, 28: 227–236.\n'},{id:"B63",body:'\nLiu T, Tang Q, Hendricks RL Inflammatory infiltration of the trigeminal ganglion after herpes simplex virus type 1 corneal infection. J Virol 1996, 70: 264–271 (1996).\n'},{id:"B64",body:'\nLiu T, Khanna KM, Chen X, Fink DJ, Hendricks RL. CD8(+) T cells can block herpes simplex virus type 1 (HSV-1) reactivation from latency in sensory neurons. J Exp Med 2000, 191: 1459–1466.\n'},{id:"B65",body:'\nTheil D, Derfuss T, Paripovic I, Herberger S, Meinl E, Schueler O, Strupp M, Arbusow V, Brandt T. Latent herpesvirus infection in human trigeminal ganglia causes chronic immune response. Am J Pathol 2003, 163: 2179–2184.\n'},{id:"B66",body:'\nKnickelbein JE, Khanna KM, Yee MB, Baty CJ, Kinchington PR, Hendricks RL. Noncytotoxic lytic granule-mediated CD8+ T cell inhibition of HSV-1 reactivation from neuronal latency. Science 2008, 322: 268–271. doi: 10.1126/science.1164164.\n'},{id:"B67",body:'\nDecman V, Kinchington PR, Harvey SA, Hendricks RL. Gamma interferon can block herpes simplex virus type 1 reactivation from latency, even in the presence of late gene expression. J Virol 2005, 79: 10339–10347.\n'},{id:"B68",body:'\nMueller SN, Heath W, McLain JD, Carbone FR, Jones CM. Characterization of two TCR transgenic mouse lines specific for herpes simplex virus. Immunol Cell Biol 2002, 80: 156–163.\n'},{id:"B69",body:'\nSt Leger AJ, Peters B, Sidney J, Sette A, Hendricks RL. Defining the herpes simplex virus-specific CD8+ T cell repertoire in C57BL/6 mice. J Immunol 2011, 186: 3927–3933. doi: 10.4049/jimmunol.1003735.\n'},{id:"B70",body:'\nDerfuss T, Segerer S, Herberger S, Sinicina I, Hüfner K, Ebelt K, Knaus HG, Steiner I, Meinl E, Dornmair K, Arbusow V, Strupp M, Brandt T, Theil D. Presence of HSV-1 immediate early genes and clonally expanded T-cells with a memory effector phenotype in human trigeminal ganglia. Brain Pathol 2007, 17: 389–398.\n'},{id:"B71",body:'\nvan Velzen M, Jing L, Osterhaus AD, Sette A, Koelle DM, Verjans GM. Local CD4 and CD8 T-cell reactivity to HSV-1 antigens documents broad viral protein expression and immune competence in latently infected human trigeminal ganglia. PLoS Pathog 2013, 9: e1003547. doi: 10.1371/journal.ppat.1003547.\n'},{id:"B72",body:'\nEgan KP, Wu S, Wigdahl B, Jennings SR. Immunological control of herpes simplex virus infections. J Neurovirol 2013, 19: 328–345. doi: 10.1007/s13365-013-0189-3.\n'},{id:"B73",body:'\nKern A, Schiff B. Vaccine therapy in recurrent herpes simplex. Arch Dermatol 1964, 89: 844–845.\n'},{id:"B74",body:'\nKoelle D. HSV 1 and 2 immunobiology and host response, pp. 616–641, In: Arvin A, Campadelli-Fiume G, Mocarski E, Moore PS, Roizman B, Whitley R, Yamanishi K (eds). Human Herpesviruses: Biology, Therapy and Immunoprophylaxis. Cambridge University Press, 2007.\n'},{id:"B75",body:'\nSkinner GR, Buchan A, Hartley CE, Turner SP, Williams DR. The preparation, efficacy and safety of \'antigenoid\' vaccine NFU1 (S-L+) MRC toward prevention of herpes simplex virus infections in human subjects. Med Microbiol Immunol 1980, 169: 39–51.\n'},{id:"B76",body:'\nDundarov S, Andonov P, Bakalov B, Nechev K, Tomov C. Immunotherapy with inactivated polyvalent herpes vaccines. Dev Biol Stand 1982, 52: 351–358.\n'},{id:"B77",body:'\nScriba M. Animal studies on the efficacy of vaccination against recurrent herpes. Med Microbiol Immunol 1982, 171: 33–40.\n'},{id:"B78",body:'\nKutinová V, Vonka V, Řezáčová D. Production and some properties of antigen extracts for neutralizing of herpes simplex virus. Acta Virol 1977, 19: 189–197.\n'},{id:"B79",body:'\nRajčáni J, Kutinová L, Vonka V. Restriction of latent herpes virus infection of rabbits immunized with subviral herpes simplex virus vaccine. Acta Virol 1980, 24: 183–193.\n'},{id:"B80",body:'\nMorello CS, Kraynyak KA, Levinson MS, Chen Z, Lee KF, Spector DH. Inactivated HSV-2 in MPL/alum adjuvant provides nearly complete protection against genital infection and shedding following long term challenge and rechallenge. Vaccine 2012, 30: 6541–6550. doi: 10.1016/j.vaccine.2012.08.049.\n'},{id:"B81",body:'\nDundarov S, Andonov P. Seventeen years of application of herpes vaccines in Bulgaria. Acta Virol 1994, 38: 205–208.\n'},{id:"B82",body:'\nHilfenhaus J, Christ H, Köhler R, Moser H, Kirchner H, Levy HB. Protectivity of herpes simplex virus antigens: studies in mice on the adjuvant effect of PICLC and on the dependence of protection on T cell competence. Med Microbiol Immunol 1981, 169: 225–235.\n'},{id:"B83",body:'\nThomson TA, Hilfenhaus J, Moser H, Morahan PS. Comparison of effects of adjuvants on efficacy of virion envelope herpes simplex virus vaccine against labial infection of BALB/c mice. Infect Immun 1983, 41: 556–562.\n'},{id:"B84",body:'\nMetcalf JF, Whitley RJ. Protective immunity against herpetic ocular disease in an outbred mouse model. Curr Eye Res 1987, 6: 167–171.\n'},{id:"B85",body:'\nRajčáni J, Sabó A, Mucha V, Košťál M, Compel P. Herpes simplex virus type 1 envelope subunit vaccine not only protects against lethal virus challenge, but also may restrict latency and virus reactivation. Acta Virol 1995, 39: 37–49.\n'},{id:"B86",body:'\nMyers MG, Bernstein DI, Harrison CJ, Stanberry LR. Herpes simplex virus glycoprotein treatment of recurrent genital herpes reduces cervicovaginal virus shedding in guinea pigs. Antiviral Res 1988, 10: 83–88.\n'},{id:"B87",body:'\nBlacklaws BA, Krishna S, Minson AC, Nash AA. Immunogenicity of herpes simplex virus type 1 glycoproteins expressed in vaccinia virus recombinants. Virology 1990, 177: 727–736.\n'},{id:"B88",body:'\nYork LJ, Giorgio DP, Mishkin EM. Immunomodulatory effects of HSV2 glycoprotein D in HSV1 infected mice: implications for immunotherapy of recurrent HSV infection. Vaccine 1995, 13: 1706–1712.\n'},{id:"B89",body:'\nMosko T, Kosovský J, Rezuchová I, Durmanová V, Kúdelová M, Rajcáni J. Expression of herpes simplex virus 1 glycoprotein D in prokaryotic and eukaryotic cells. Acta Virol 2004, 48: 97–107.\n'},{id:"B90",body:'\nDurmanová V, Mosko T, Sapák M, Kosovský J, Rezuchová I, Buc M, Rajcáni J. Efficacy of recombinant herpes simplex virus 1 glycoprotein D candidate vaccines in mice. Acta Microbiol Immunol Hung 2006, 53: 459–477.\n'},{id:"B91",body:'\nLong D, Madara TJ, Ponce de Leon M, Cohen GH, Montgomery PC, Eisenberg RJ. Glycoprotein D protects mice against lethal challenge with herpes simplex virus types 1 and 2. Infect Immun 1984, 43: 761–764.\n'},{id:"B92",body:'\nRoberts PL, Duncan BE, Raybould TJ, Watson DH. Purification of herpes simplex virus glycoproteins B and C using monoclonal antibodies and their ability to protect mice against lethal challenge. J Gen Virol 1985, 66: 1073–1085.\n'},{id:"B93",body:'\nKino Y, Eto T, Nishiyama K, Ohtomo N, Mori R. Immunogenicity of purified glycoprotein gB of herpes simplex virus. Arch Virol 1986, 89: 69–80.\n'},{id:"B94",body:'\nManservigi R, Grossi MP, Gualandri R, Balboni PG, Marchini A, Rotola A, Rimessi P, Di Luca D, Cassai E, Barbanti-Brodano G. Protection from herpes simplex virus type 1 lethal and latent infections by secreted recombinant glycoprotein B constitutively expressed in human cells with a BK virus episomal vector. J Virol 1990, 64: 431–436.\n'},{id:"B95",body:'\nBroker M, Abel KJ, Köhler R, Hilfenhaus J, Amann E. Escherichia coli-derived envelope protein gD but not gC antigens of herpes simplex virus protect mice against a lethal challenge with HSV-1 and HSV-2. Med Microbiol Immunol 1990, 179: 145–159.\n'},{id:"B96",body:'\nByars NE, Fraser-Smith EB, Pecyk RA, Welch M, Nakano G, Burke RL, Hayward AR, Allison AC. Vaccinating guinea pigs with recombinant glycoprotein D of herpes simplex virus in an efficacious adjuvant formulation elicits protection against vaginal infection. Vaccine 1994, 12: 200–209.\n'},{id:"B97",body:'\nO\'Hagan D, Goldbeck C, Ugozzoli M, Ott G, Burke RL. Intranasal immunization with recombinant gD2 reduces disease severity and mortality following genital challenge with herpes simplex virus type 2 in guinea pigs. Vaccine 1999, 17: 2229–2236.\n'},{id:"B98",body:'\nBourne N, Bravo FJ, Francotte M, Bernstein DI, Myers MG, Slaoui M, Stanberry LR. Herpes simplex virus (HSV) type 2 glycoprotein D subunit vaccines and protection against genital HSV-1 or HSV-2 disease in guinea pigs. J Infect Dis 2003, 187: 542–549.\n'},{id:"B99",body:'\nManservigi R, Boero A, Argnani R, Caselli E, Zucchini S, Miriagou V, Mavromara P, Cilli M, Grossi MP, Balboni PG, Cassai E. Immunotherapeutic activity of a recombinant combined gB-gD-gE vaccine against recurrent HSV-2 infections in a guinea pig model. Vaccine 2005, 23: 865–872.\n'},{id:"B100",body:'\nDurmanová V, Sapák M, Kosovský J, Rezuchová I, Kúdelová M, Buc M, Rajcáni J. Immune response and cytokine production following immunization with experimental herpes simplex virus 1 (HSV-1) vaccines. Folia Microbiol (Praha) 2008, 53: 73–83. doi: 10.1007/s12223-008-0011-4.\n'},{id:"B101",body:'\nFotouhi F, Soleimanjahi H, Roostaee MH, Dalimi Asl A. Expression of the herpes simplex virus type 2 glycoprotein D in baculovirus expression system and evaluation of its immunogenicity in guinea pigs. Iran Biomed J 2008, 12: 59–66.\n'},{id:"B102",body:'\nAwasthi S, Balliet JW, Flynn JA, Lubinski JM, Shaw CE, DiStefano DJ, Cai M, Brown M, Smith JF, Kowalski R, Swoyer R, Galli J, Copeland V, Rios S, Davidson RC, Salnikova M, Kingsley S, Bryan J, Casimiro DR, Friedman HM. Protection provided by a herpes simplex virus 2 (HSV-2) glycoprotein C and D subunit antigen vaccine against genital HSV-2 infection in HSV-1-seropositive guinea pigs. J Virol 2014, 88: 2000–2010. doi: 10.1128/JVI.03163-13.\n'},{id:"B103",body:'\nAwasthi S, Mahairas GG, Shaw CE, Huang ML, Koelle DM, Posavad C, Corey L, Friedman HM. A dual-modality herpes simplex virus 2 vaccine for preventing genital herpes by using glycoprotein C and D subunit antigens to induce potent antibody responses and adenovirus vectors containing capsid and tegument proteins as T cell immunogens. J Virol 2015, 89: 8497–8509. doi: 10.1128/JVI.01089-15.\n'},{id:"B104",body:'\nInoue T, Inoue Y, Nakamura T, Yoshida A, Inoue Y, Tano Y, Shimomura Y, Fujisawa Y, Aono A, Hayashi K. The effect of immunization with herpes simplex virus glycoprotein D fused with interluekin-2 against murine herpetic keratitis. Nihon Ganka Gakkai Zasshi 2001, 105: 223–229.\n'},{id:"B105",body:'\nMohamedi SA, Heath AW, Jennings R. A comparison of oral and parenteral routes for therapeutic vaccination with HSV-2 ISCOMs in mice; cytokine profiles, antibody responses and protection. Antiviral Res 2001, 49: 83–99.\n'},{id:"B106",body:'\nBurke RL, Goldbeck C, Ng P, Stanberry L, Ott G, Van Nest G. The influence of adjuvant on the therapeutic efficacy of a recombinant genital herpes vaccine. J Infect Dis 1994, 170: 1110–1119.\n'},{id:"B107",body:'\nShoenfeld Y, Agmont-Levine N. ASIA: autoimmune/inflammatory syndrome induced by adjuvants. J Autoimmun 2011, 36: 4–8. doi: 10.1016/j.jaut.2010.07.003.\n'},{id:"B108",body:'\nLavelle EC, Grant G, Pusztai A, Pfüller U, Leavy O, McNeela E, Mills KH, O\'Hagan DT. Mistletoe lectins enhance immune responses to intranasally co-administered herpes simplex virus glycoprotein D2. Immunology 2002, 107: 268–274.\n'},{id:"B109",body:'\nCortesi R, Ravani L, Rinaldi F, Marconi P, Drechsler M, Manservigi M, Argnani R, Menegatti E, Esposito E, Manservigi R. Intranasal immunization in mice with non-ionic surfactants vesicles containing HSV immunogens: a preliminary study as possible vaccine against genital herpes. Int J Pharm 2013, 440: 229–237. doi: 10.1016/j.ijpharm.2012.06.042.\n'},{id:"B110",body:'\nSkoberne M, Cardin R, Lee A, Kazimirova A, Zielinski V, Garvie D, Lundberg A, Larson S, Bravo FJ, Bernstein DI, Flechtner JB, Long D. An adjuvanted herpes simplex virus 2 subunit vaccine elicits a T cell response in mice and is an effective therapeutic vaccine in guinea pigs. J Virol 2013, 87: 3930–3942. doi: 10.1128/JVI.02745-12.\n'},{id:"B111",body:'\nRajčáni J, MoškoT, Režuchová, I. Current developments in viral DNA vaccines: shall they solve the unsolved? Rev Med Microbiol 2005, 15, 1–23.\n'},{id:"B112",body:'\nCantin EM, Eberle R, Baldick JL, Moss B, Willey DE, Notkins AL, Openshaw H. Expression of herpes simplex virus 1 glycoprotein B by a recombinant vaccinia virus and protection of mice against lethal herpes simplex virus 1 infection. Proc Natl Acad Sci U S A 1987, 84: 5908–5912.\n'},{id:"B113",body:'\nWilley DE, Cantin EM, Hill LR, Moss B, Notkins AL, Openshaw H. Herpes simplex virus type 1-vaccinia virus recombinant expressing glycoprotein B: protection from acute and latent infection. J Infect Dis 1988, 158: 1382–1386.\n'},{id:"B114",body:'\nWachsman M, Aurelian L, Smith CC, Perkus ME, Paoletti E. Regulation of expression of herpes simplex virus (HSV) glycoprotein D in vaccinia recombinants affects their ability to protect from cutaneous HSV-2 disease. J Infect Dis 1989, 159: 625–634.\n'},{id:"B115",body:'\nBernstein DI. Effect of route of vaccination with vaccinia virus expressing HSV-2 glycoprotein D on protection from genital HSV-2 infection. Vaccine 2000, 18: 1351–1358.\n'},{id:"B116",body:'\nNatuk RJ, Cooper D, Guo M, Calderon P, Wright KJ, Nasar F, Witko S, Pawlyk D, Lee M, DeStefano J, Tummolo D, Abramovitz AS, Gangolli S, Kalyan N, Clarke DK, Hendry RM, Eldridge JH, Udem SA, Kowalski J. Recombinant vesicular stomatitis virus vectors expressing herpes simplex virus type 2 gD elicit robust CD4+ Th1 immune responses and are protective in mouse and guinea pig models of vaginal challenge. J Virol 2006, 80: 4447–4457.\n'},{id:"B117",body:'\nChiuppesi F, Vannucci L, De Luca A, Lai M, Matteoli B, Freer G, Manservigi R, Ceccherini-Nelli L, Maggi F, Bendinelli M, Pistello M. A lentiviral vector-based, herpes simplex virus 1 (HSV-1) glycoprotein B vaccine affords cross-protection against HSV-1 and HSV-2 genital infections. J Virol 2012, 86: 6563–6574. doi: 10.1128/JVI.00302-12.\n'},{id:"B118",body:'\nDaheshia M, Kuklin N, Manickan E, Chun S, Rouse BT. Immune induction and modulation by topical ocular administration of plasmid DNA encoding antigens and cytokines. Vaccine 1998, 16: 1103–1110.\n'},{id:"B119",body:'\nNass PH, Elkins KL, Weir JP. Antibody response and protective capacity of plasmid vaccines expressing three different herpes simplex virus glycoproteins. J Infect Dis 1998, 178: 611–617.\n'},{id:"B120",body:'\nStanberry LR, Cunningham AL, Mindel A, Scott LL, Spruance SL, Aoki FY, Lacey CJ. Prospects for control of herpes simplex virus disease through immunization. Clin Infect Dis 2000, 30: 549–566.\n'},{id:"B121",body:'\nBaghian A, Chouljenko VN, Dauvergne O, Newmant MJ, Baghian S, Kousoulas KG. Protective immunity against lethal HSV-1 challenge in mice by nucleic acid-based immunisation with herpes simplex virus type-1 genes specifying glycoproteins gB and gD. J Med Microbiol 2002, 51: 350–357.\n'},{id:"B122",body:'\nOsorio Y, Cohen J, Ghiasi H. Improved protection from primary ocular HSV-1 infection and establishment of latency using multigenic DNA vaccines. Invest Ophthalmol Vis Sci 2004, 45: 506–514\n'},{id:"B123",body:'\nHoshino Y, Dalai SK, Wang K, Pesnicak L, Lau TY, Knipe DM, Cohen JI, Straus SE. Comparative efficacy and immunogenicity of replication-defective, recombinant glycoprotein, and DNA vaccines for herpes simplex virus 2 infections in mice and guinea pigs. J Virol 2005, 79: 410–418.\n'},{id:"B124",body:'\nGhaemi A, Soleimanjahi H, Bamdad T, Soudi S, Arefeian E, Hashemi SM, Ebtekar M. Induction of humoral and cellular immunity against latent HSV-1 infections by DNA immunization in BALB/c mice. Comp Immunol Microbiol Infect Dis 2007, 30: 197–210.\n'},{id:"B125",body:'\nBourne N, Milligan GN, Schleiss MR, Bernstein DI, Stanberry LR. DNA immunization confers protective immunity on mice challenged intravaginally with herpes simplex virus type 2. Vaccine 1996, 14: 1230–1234.\n'},{id:"B126",body:'\nMcClements WL, Armstrong ME, Keys RD, Liu MA. The prophylactic effect of immunization with DNA encoding herpes simplex virus glycoproteins on HSV-induced disease in guinea pigs. Vaccine 1997, 15: 857–860.\n'},{id:"B127",body:'\nStrasser JE, Arnold RL, Pachuk C, Higgins TJ, Bernstein DI. Herpes simplex virus DNA vaccine efficacy: effect of glycoprotein D plasmid constructs. J Infect Dis 2000, 182: 1304–1310.\n'},{id:"B128",body:'\nJazayeri M, Soleimanjahi H, Fotouhi F, Pakravan N. Comparison of intramuscular and footpad subcutaneous immunization with DNA vaccine encoding HSV-gD2 in mice. Comp Immunol Microbiol Infect Dis 2009, 32: 453–461. doi: 10.1016/j.cimid.2008.05.002.\n'},{id:"B129",body:'\nLee HH, Cha SC, Jang DJ, Lee JK, Choo DW, Kim YS, Uh HS, Kim SY. Immunization with combined HSV-2 glycoproteins B2: D2 gene DNAs: protection against lethal intravaginal challenges in mice. Virus Genes 2002, 25: 179–188.\n'},{id:"B130",body:'\nDomingo C, Gadea I, Pardeiro M, Castilla C, Fernández S, Fernández-Clua MA, De la Cruz Troca JJ, Punzón C, Soriano F, Fresno M, Tabarés E. Immunological properties of a DNA plasmid encoding a chimeric protein of herpes simplex virus type 2 glycoprotein B and glycoprotein D. Vaccine 2003, 21: 3565–3574\n'},{id:"B131",body:'\nSin JI, Kim JJ, Arnold RL, Shroff KE, McCallus D, Pachuk C, McElhiney SP, Wolf MW, Pompa-de Bruin SJ, Higgins TJ, Ciccarelli RB, Weiner DB. IL-12 gene as a DNA vaccine adjuvant in a herpes mouse model: IL-12 enhances Th1-type CD4+ T cell-mediated protective immunity against herpes simplex virus-2 challenge. J Immun 1999, 162: 2912–2921.\n'},{id:"B132",body:'\nCooper D, Pride MW, Guo M, Cutler M, Mester JC, Nasar F, She J, Souza V, York L, Mishkin E, Eldridge J, Natuk RJ. Interleukin-12 redirects murine immune responses to soluble or aluminum phosphate adsorbed HSV-2 glycoprotein D towards Th1 and CD4+ CTL responses. Vaccine 2004, 23: 236–246.\n'},{id:"B133",body:'\nSin JI, Kim JJ, Boyer JD, Ciccarelli RB, Higgins TJ, Weiner DB. In vivo modulation of vaccine-induced immune responses toward a Th1 phenotype increases potency and vaccine effectiveness in a herpes simplex virus type 2 mouse model. J Virol 1999, 73: 501–509.\n'},{id:"B134",body:'\nZhu M, Xu X, Liu H, Liu X, Wang S, Dong F, Yang B, Song G. Enhancement of DNA vaccine potency against herpes simplex virus 1 by co-administration of an interleukin-18 expression plasmid as a genetic adjuvant. J Med Microbiol 2003, 52: 223–228.\n'},{id:"B135",body:'\nOsorio Y, Ghiasi H. Comparison of adjuvant efficacy of herpes simplex virus type 1 recombinant viruses expressing TH1 and TH2 cytokine genes. J Virol 2003, 77: 5774–5783.\n'},{id:"B136",body:'\nLee S, Gierynska M, Eo SK, Kuklin N, Rouse BT. Influence of DNA encoding cytokines on systemic and mucosal immunity following genetic vaccination against herpes simplex virus. Microbes Infect 2003, 5: 571–578.\n'},{id:"B137",body:'\nVeselenak RL, Shlapobersky M, Pyles RB, Wei Q, Sullivan SM, Bourne N. A Vaxfectin-adjuvanted HSV-2 plasmid DNA vaccine is effective for prophylactic and therapeutic use in the guinea pig model of genital herpes. Vaccine 2012, 30: 7046–7051. doi: 10.1016/j.vaccine.2012.09.057.\n'},{id:"B138",body:'\nShlapobersky M, Marshak JO, Dong L, Huang ML, Wei Q, Chu A, Rolland A, Sullivan S, Koelle DM. Vaxfectin-adjuvanted plasmid DNA vaccine improves protection and immunogenicity in a murine model of genital herpes infection. J Gen Virol 2012, 93: 1305–1315. doi: 10.1099/vir.0.040055-0.\n'},{id:"B139",body:'\nYan Y, Hu K, Deng X, Guan X, Luo S, Tong L, Du T, Fu M, Zhang M, Liu Y, Hu Q. Immunization with HSV-2 gB-CCL19 Fusion Constructs Protects Mice against Lethal Vaginal Challenge. J Immunol 2015, 195: 329–338. doi: 10.4049/jimmunol.1500198.\n'},{id:"B140",body:'\nBright H, Perez DL, Christy C, Cockle P, Eyles JE, Hammond D, Khodai T, Lang S, West K, Loudon PT. The efficacy of HSV-2 vaccines based on gD and gB is enhanced by the addition of ICP27. Vaccine 2012, 30: 7529–7535. doi: 10.1016/j.vaccine.2012.10.046.\n'},{id:"B141",body:'\nForrester A, Farrell H, Wilkinson G, Kaye J, Davis-Poynter N, Minson T. Construction and properties of a mutant of herpes simplex virus type 1 with glycoprotein H coding sequences deleted. J Virol 1992, 66: 341–348.\n'},{id:"B142",body:'\nMcLean CS, Erturk M, Jennings R, Challanain DN, Minson AC, Duncan I, Boursnell ME, Inglis SC. Protective vaccination against primary and recurrent disease caused by herpes simplex virus (HSV) type 2 using a genetically disabled HSV-1. J Infect Dis 1994, 170: 1100–1109.\n'},{id:"B143",body:'\nWhitley RJ, Kern ER, Chatterjee S, Chou J, Roizman B. Replication, establishment of latency, and induced reactivation of herpes simplex virus gamma 1 34.5 deletion mutants in rodent models. J Clin Invest 1993, 91: 2837–2843.\n'},{id:"B144",body:'\nBrittle EE, Wang F, Lubinski JM, Bunte RM, Friedman HM. A replication-competent, neuronal spread-defective, live attenuated herpes simplex virus type 1 vaccine. J Virol 2008, 82: 8431–8441. doi: 10.1128/JVI.00551-08.\n'},{id:"B145",body:'\nAwasthi S, Zumbrun EE, Si H, Wang F, Shaw CE, Cai M, Lubinski JM, Barrett SM, Balliet JW, Flynn JA, Casimiro DR, Bryan JT, Friedman HM. Live attenuated herpes simplex virus 2 glycoprotein E deletion mutant as a vaccine candidate defective in neuronal spread. J Virol 2012, 86: 4586–4598. doi: 10.1128/JVI.07203-11.\n'},{id:"B146",body:'\nKeadle TL, Laycock KA, Morris JL, Leib DA, Morrison LA, Pepose JS, Stuart PM. Therapeutic vaccination with vhs(-) herpes simplex virus reduces the severity of recurrent herpetic stromal keratitis in mice. J Gen Virol 2002, 83: 2361–2365.\n'},{id:"B147",body:'\nRajčáni J, Kúdelová M, Oravcová I, Vojvodová A, Košovský J, Matis J. Characterization of strain HSZP of herpes simplex virus type 1 (HSV1). Folia Microbiol (Praha) 1999, 44: 713–719.\n'},{id:"B148",body:'\nMeignier B. Genetically engineered attenuated herpes simplex viruses. Rev Infect Dis 1991, 13: S895–897.\n'},{id:"B149",body:'\nHalford WP, Püschel R, Gershburg E, Wilber A, Gershburg S, Rakowski B. A live-attenuated HSV-2 ICP0 virus elicits 10 to 100 times greater protection against genital herpes than a glycoprotein D subunit vaccine. PLoS One 2011, 6: e17748. doi: 10.1371/journal.pone.0017748.\n'},{id:"B150",body:'\nde Bruyn G, Vargas-Cortez M, Warren T, Tyring SK, Fife KH, Lalezari J, Brady RC, Shahmanesh M, Kinghorn G, Beutner KR, Patel R, Drehobl MA, Horner P, Kurtz TO, McDermott S, Wald A, Corey L. A randomized controlled trial of a replication defective (gH deletion) herpes simplex virus vaccine for the treatment of recurrent genital herpes among immunocompetent subjects. Vaccine 2006, 24: 914–920.\n'},{id:"B151",body:'\nMorrison LA, Knipe DM. Immunization with replication-defective mutants of herpes simplex virus type 1: sites of immune intervention in pathogenesis of challenge virus infection. J Virol 1994, 68: 689–696.\n'},{id:"B152",body:'\nWachsman M, Kulka M, Smith CC, Aurelian L. A growth and latency compromised herpes simplex virus type 2 mutant (ICP10DeltaPK) has prophylactic and therapeutic protective activity in guinea pigs. Vaccine 2001, 19: 1879–1890.\n'},{id:"B153",body:'\nDa Costa XJ, Bourne N, Stanberry LR, Knipe DM. Construction and characterization of a replication-defective herpes simplex virus 2 ICP8 mutant strain and its use in immunization studies in a guinea pig model of genital disease. Virology 1997, 232: 1–12.\n'},{id:"B154",body:'\nDa Costa X, Kramer MF, Zhu J, Brockman MA, Knipe DM. Construction, phenotypic analysis, and immunogenicity of a UL5/UL29 double deletion mutant of herpes simplex virus 2. J Virol 2000, 74: 7963–7971.\n'},{id:"B155",body:'\nDelagrave S, Hernandez H, Zhou C, Hamberger JF, Mundle ST, Catalan J, Baloglu S, Anderson SF, DiNapoli JM, Londoño-Hayes P, Parrington M, Almond J, Kleanthous H. Immunogenicity and efficacy of intramuscular replication-defective and subunit vaccines against herpes simplex virus type 2 in the mouse genital model. PLoS One 2012, 7: e46714. doi: 10.1371/journal.pone.0046714.\n'},{id:"B156",body:'\nBrans R, Yao F. Immunization with a dominant-negative recombinant Herpes Simplex Virus (HSV) type 1 protects against HSV-2 genital disease in guinea pigs. BMC Microbiol 2010, 10: 163. doi: 10.1186/1471-2180-10-163.\n'},{id:"B157",body:'\nCasanova G, Cancela R, Alonzo L, Benuto R, Magana Mdel C, Hurley DR, Fishbein E, Lara C, Gonzalez T, Ponce R, Burnett JW, Calton GJ. A double-blind study of the efficacy and safety of the ICP10deltaPK vaccine against recurrent genital HSV-2 infections. Cutis 2002, 70: 235–239.\n'},{id:"B158",body:'\nvan Lint AL, Torres-Lopez E, Knipe DM. Immunization with a replication-defective herpes simplex virus 2 mutant reduces herpes simplex virus 1 infection and prevents ocular disease. Virology 2007, 368: 227–231.\n'},{id:"B159",body:'\nHoshino Y, Pesnicak L, Dowdell KC, Burbelo PD, Knipe DM, Straus SE, Cohen JI. Protection from herpes simplex virus (HSV)-2 infection with replication-defective HSV-2 or glycoprotein D2 vaccines in HSV-1-seropositive and HSV-1-seronegative guinea pigs. J Infect Dis 2009, 200: 1088–1095. doi: 10.1086/605645.\n'},{id:"B160",body:'\nStanfield BA, Stahl J, Chouljenko VN, Subramanian R, Charles AS, Saied AA, Walker JD, Kousoulas KG. A single intramuscular vaccination of mice with the HSV-1 VC2 virus with mutations in the glycoprotein K and the membrane protein UL20 confers full protection against lethal intravaginal challenge with virulent HSV-1 and HSV-2 strains. PLoS One 2014, 9:e109890. doi: 10.1371/journal.pone.0109890.\n'},{id:"B161",body:'\nZhang X, Castelli FA, Zhu X, Wu M, Maillère B, BenMohamed L. Gender-dependent HLA-DR-restricted epitopes identified from herpes simplex virus type 1 glycoprotein D. Clin Vaccine Immunol 2008, 15: 1436–1449. doi: 10.1128/CVI.00123-08.\n'},{id:"B162",body:'\nMertz GJ, Peterman G, Ashley R, Jourden JL, Salter D, Morrison L, McLean A, Corey L. Herpes simplex virus type-2 glycoprotein-subunit vaccine: tolerance and humoral and cellular responses in humans. J Infect Dis 1984, 150: 242–249.\n'},{id:"B163",body:'\nMertz GJ, Ashley R, Burke RL, Benedetti J, Critchlow C, Jones CC, Corey L. Double-blind, placebo-controlled trial of a herpes simplex virus type 2 glycoprotein vaccine in persons at high risk for genital herpes infection. J Infect Dis 1990, 161: 653–660\n'},{id:"B164",body:'\nLangenberg AG, Burke RL, Adair SF, Sekulovich R, Tigges M, Dekker CL, Corey L. A recombinant glycoprotein vaccine for herpes simplex virus type 2: safety and immunogenicity. Ann Intern Med 1995, 122: 889–898.\n'},{id:"B165",body:'\nBernstein DI, Aoki FY, Tyring SK, Stanberry LR, St-Pierre C, Shafran SD, Leroux-Roels G, Van Herck K, Bollaerts A, Dubin G; GlaxoSmithKline Herpes Vaccine Study Group. Safety and immunogenicity of glycoprotein D-adjuvant genital herpes vaccine. Clin Infect Dis 2005, 40: 1271–1281.\n'},{id:"B166",body:'\nStanberry LR, Spruance SL, Cunningham AL, Bernstein DI, Mindel A, Sacks S, Tyring S, Aoki FY, Slaoui M, Denis M, Vandepapeliere P, Dubin G; GlaxoSmithKline Herpes Vaccine Efficacy Study Group. Glycoprotein-D-adjuvant vaccine to prevent genital herpes. N Engl J Med 2002, 347: 1652–1661.\n'},{id:"B167",body:'\nBelshe RB, Leone PA, Bernstein DI, Wald A, Levin MJ, Stapleton JT, Gorfinkel I, Morrow RL, Ewell MG, Stokes-Riner A, Dubin G, Heineman TC, Schulte JM, Deal CD; Herpevac Trial for Women. Efficacy results of a trial of a herpes simplex vaccine. N Engl J Med 2012, 366: 34–43. doi: 10.1056/NEJMoa1103151.\n'},{id:"B168",body:'\nTavares F, Cheuvart B, Heineman T, Arellano F, Dubin G. Meta-analysis of pregnancy outcomes in pooled randomized trials on a prophylactic adjuvanted glycoprotein D subunit herpes simplex virus vaccine. Vaccine 2013, 31: 1759–1764. doi: 10.1016/j.vaccine.2013.01.002.\n'},{id:"B169",body:'\nCattamanchi A, Posavad CM, Wald A, Baine Y, Moses J, Higgins TJ, Ginsberg R, Ciccarelli R, Corey L, Koelle DM. Phase I study of a herpes simplex virus type 2 (HSV-2) DNA vaccine administered to healthy, HSV-2-seronegative adults by a needle-free injection system. Clin Vaccine Immunol 2008, 15: 1638–1643. doi: 10.1128/CVI.00167-08.\n'},{id:"B170",body:'\nBernard MC, Barban V, Pradezynski F, de Montfort A, Ryall R, Caillet C, Londono-Hayes P. Immunogenicity, protective efficacy, and non-replicative status of the HSV-2 vaccine candidate HSV529 in mice and guinea pigs. PLoS One 2015, 10: e0121518. doi: 10.1371/journal.pone.0121518.\n'},{id:"B171",body:'\nKutinová L, Benda R, Kalos Z, Dbalý V, Votruba T, Kvícalová E, Petrovská P, Doutlík S, Kamínková J, Domorázková E. Placebo-controlled study with subunit herpes simplex virus vaccine in subjects suffering from frequent herpetic recurrences. Vaccine 1988, 6: 223–228.\n'},{id:"B172",body:'\nSkinner GR, Turyk ME, Benson CA, Wilbanks GD, Heseltine P, Galpin J, Kaufman R, Goldberg L, Hartley CE, Buchan A. The efficacy and safety of Skinner herpes simplex vaccine towards modulation of herpes genitalis; report of a prospective double-blind placebo-controlled trial. Med Microbiol Immunol 1997, 186: 31–36.\n'},{id:"B173",body:'\nStraus SE, Corey L, Burke RL, Savarese B, Barnum G, Krause PR, Kost RG, Meier JL, Sekulovich R, Adair SF. Placebo-controlled trial of vaccination with recombinant glycoprotein D of herpes simplex virus type 2 for immunotherapy of genital herpes. Lancet 1994, 343: 1460–1463.\n'},{id:"B174",body:'\nStraus SE, Wald A, Kost RG, McKenzie R, Langenberg AG, Hohman P, Lekstrom J, Cox E, Nakamura M, Sekulovich R, Izu A, Dekker C, Corey L. Immunotherapy of recurrent genital herpes with recombinant herpes simplex virus type 2 glycoproteins D and B: results of a placebo-controlled vaccine trial. J Infect Dis 1997, 176: 1129–1134.\n'},{id:"B175",body:'\nLaing KJ, Dong L, Sidney J, Sette A, Koelle DM. Immunology in the Clinic Review Series; focus on host responses: T cell responses to herpes simplex viruses. Clin Exp Immunol 2012, 167: 47–58. doi: 10.1111/j.1365-2249.2011.04502.x\n'},{id:"B176",body:'\nLaing KJ, Magaret AS, Mueller DE, Zhao L, Johnston C, De Rosa SC, Koelle DM, Wald A, Corey L. Diversity in CD8(+) T cell function and epitope breadth among persons with genital herpes. J Clin Immun 2010, 30: 703–722. doi: 10.1007/s10875-010-9441-2.\n'},{id:"B177",body:'\nBerger C, Flowers ME, Warren EH, Riddell SR. Analysis of transgene-specific immune responses that limit the in vivo persistence of adoptively transferred HSV-TK-modified donor T cells after allogeneic hematopoietic cell transplantation. Blood 2006, 107: 2294–2302.\n'},{id:"B178",body:'\nKoelle DM, Liu Z, McClurkan CL, Cevallos RC, Vieira J, Hosken NA, Meseda CA, Snow DC, Wald A, Corey L. Immunodominance among herpes simplex virus-specific CD8 T cells expressing a tissue-specific homing receptor. Proc Natl Acad Sci U S A 2003, 100: 12899–12904.\n'},{id:"B179",body:'\nChentoufi AA, Binder NR, Berka N, Durand G, Nguyen A, Bettahi I, Maillère B, BenMohamed L. Asymptomatic human CD4+ cytotoxic T-cell epitopes identified from herpes simplex virus glycoprotein B. J Virol 2008, 82: 11792–11802. doi: 10.1128/JVI.00692-08\n'},{id:"B180",body:'\nWang X, Xie G, Liao J, Yin D, Guan W, Pan M, Li J, Li Y. Design and evaluation of a multi-epitope assembly peptide (MEAP) against herpes simplex virus type 2 infection in BALB/c mice. Virol J 2011, 8: 232. doi: 10.1186/1743-422X-8-232.\n'},{id:"B181",body:'\nLiu K, Jiang D, Zhang L, Yao Z, Chen Z, Yu S, Wang X. Identification of B- and T-cell epitopes from glycoprotein B of herpes simplex virus 2 and evaluation of their immunogenicity and protection efficacy. Vaccine 2012, 30: 3034–3041. doi: 10.1016/j.vaccine.2011.10.010.\n'},{id:"B182",body:'\nMuller WJ, Dong L, Vilalta A, Byrd B, Wilhelm KM, McClurkan CL, Margalith M, Liu C, Kaslow D, Sidney J, Sette A, Koelle DM. Herpes simplex virus type 2 tegument proteins contain subdominant T-cell epitopes detectable in BALB/c mice after DNA immunization and infection. J Gen Virol 2009, 90: 1153–1163. doi: 10.1099/vir.0.008771-0.\n'},{id:"B183",body:'\nKoelle DM, Frank JM, Johnson ML, Kwok WW. Recognition of herpes simplex virus type 2 tegument proteins by CD4 T cells infiltrating human genital herpes lesions. J Virol 1998, 72: 7476–7483.\n'},{id:"B184",body:'\nNovak EJ, Liu AW, Gebe JA, Falk BA, Nepom GT, Koelle DM, Kwok WW. Tetramer-guided epitope mapping: rapid identification and characterization of immunodominant CD4+ T cell epitopes from complex antigens. J Immunol 2001, 166: 6665–6670.\n'},{id:"B185",body:'\nOsorio Y, Mott KR, Jabbar AM, Moreno A, Foster TP, Kousoulas KG, Ghiasi H. Epitope mapping of HSV-1 glycoprotein K (gK) reveals a T cell epitope located within the signal domain of gK. Virus Res 2007, 128: 71–80.\n'},{id:"B186",body:'\nMott KR, Chentoufi AA, Carpenter D, BenMohamed L, Wechsler SL, Ghiasi H. The role of a glycoprotein K (gK) CD8+ T-cell epitope of herpes simplex virus on virus replication and pathogenicity. Invest Ophthalmol Vis Sci 2009, 50: 2903–2912. doi: 10.1167/iovs.08-2957.\n'},{id:"B187",body:'\nHaynes JR, Arrington J, Dong L, Braun RP, Payne LG. Potent protective cellular immune responses generated by a DNA vaccine encoding HSV-2 ICP27 and the E. coli heat labile enterotoxin. Vaccine 2006, 24: 5016–5026.\n'},{id:"B188",body:'\nBenMohamed L, Bertrand G, McNamara CD, Gras-Masse H, Hammer J, Wechsler SL, Nesburn AB. Identification of novel immunodominant CD4+ Th1-type T-cell peptide epitopes from herpes simplex virus glycoprotein D that confer protective immunity. J Virol 2003, 77: 9463–9473.\n'},{id:"B189",body:'\nCooper D, Mester JC, Guo M, Nasar F, Souza V, Dispoto S, Sidhu M, Hagen M, Eldridge JH, Natuk RJ, Pride MW. Epitope mapping of full-length glycoprotein D from HSV-2 reveals a novel CD4+ CTL epitope located at the transmembrane-cytoplasmic junction. Cell Immunol 2006, 239: 113–120.\n'},{id:"B190",body:'\nChentoufi AA, Zhang X, Lamberth K, Dasgupta G, Bettahi I, Nguyen A, Wu M, Zhu X, Mohebbi A, Buus S, Wechsler SL, Nesburn AB, BenMohamed L. HLA-A*02:01-restricted CD8+ cytotoxic T lymphocyte epitopes identified from herpes simplex virus glycoprotein D. J Immunol 2008, 180: 426–437.\n'},{id:"B191",body:'\nChentoufi AA, Dasgupta G, Christensen ND, Hu J, Choudhury ZS, Azeem A, Jester JV, Nesburn AB, Wechsler SL, BenMohamed L. A novel HLA (HLA-A*02:01) transgenic rabbit model for preclinical evaluation of human CD8+ T cell epitope-based vaccines against ocular herpes. J Immunol 2010, 184: 2561–2571. doi: 10.4049/jimmunol.0902322.\n'},{id:"B192",body:'\nPosavad CM, Remington M, Mueller DE, Zhao L, Magaret AS, Wald A, Corey L. Detailed characterization of T cell responses to herpes simplex virus-2 in immune seronegative persons. J Immunol 2010, 184: 3250–3259. doi: 10.4049/jimmunol.0900722.\n'},{id:"B193",body:'\nPosavad CM, Magaret AS, Zhao L, Mueller DE, Wald A, Corey L. Development of an interferon-gamma ELISPOT assay to detect human T cell responses to HSV-2. Vaccine 2011, 29: 7058–7066. doi: 10.1016/j.vaccine.2011.07.028.\n'},{id:"B194",body:'\nMaiers M, Gragert L, Klitz W. High-resolution HLA alleles and haplotypes in the United States population. Hum Immunol 2007, 68: 779–788.\n'},{id:"B195",body:'\nKoelle DM, Corey L. Herpes simplex: insights on pathogenesis and possible vaccines. Annu Rev Med 2008, 59: 381–395. doi: 10.1146/annurev.med.59.061606.095540.\n'},{id:"B196",body:'\nSamandary S, Kridane-Miledi H, Sandoval JS, Choudhury Z, Langa-Vives F, Spencer D, Chentoufi AA, Lemonnier FA, BenMohamed L. Associations of HLA-A, HLA-B and HLA-C alleles frequency with prevalence of herpes simplex virus infections and diseases across global populations: implication for the development of an universal CD8+ T-cell epitope-based vaccine. Hum Immunol 2014, 75: 715–729. doi: 10.1016/j.humimm.2014.04.016.\n'},{id:"B197",body:'\nAwasthi S, Friedman HM. Status of prophylactic and therapeutic genital herpes vaccines. Curr Opin Virol 2014, 6: 6–12. doi: 10.1016/j.coviro.2014.02.006.\n'},{id:"B198",body:'\nWald A, Zeh J, Selke S, Warren T, Ryncarz AJ, Ashley R, Krieger JN, Corey L. Reactivation of genital herpes simplex virus type 2 infection in asymptomatic seropositive persons. N Engl J Med 2000, 342: 844–850.\n'},{id:"B199",body:'\nKhan AA, Srivastava R, Spencer D, Garg S, Fremgen D, Vahed H, Lopes PP, Pham TT, Hewett C, Kuang J, Ong N, Huang L, Scarfone VM, Nesburn AB, Wechsler SL, BenMohamed L. Phenotypic and functional characterization of herpes simplex virus glycoprotein B epitope-specific effector and memory CD8+ T cells from symptomatic and asymptomatic individuals with ocular herpes. J Virol 2015, 89: 3776–3792. doi: 10.1128/JVI.03419-14.\n'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Vladimíra Ďurmanová",address:null,affiliation:'
Institute of Immunology, Faculty of Medicine, Comenius University in Bratislava, Bratislava, Slovakia
Institute of Virology, Slovak Academy of Sciences, Bratislava, Slovakia
'}],corrections:null},book:{id:"5304",title:"Herpesviridae",subtitle:null,fullTitle:"Herpesviridae",slug:"herpesviridae",publishedDate:"September 7th 2016",bookSignature:"Jozsef Ongradi",coverURL:"https://cdn.intechopen.com/books/images_new/5304.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"36151",title:"Dr.",name:"Joseph",middleName:null,surname:"Ongrádi",slug:"joseph-ongradi",fullName:"Joseph Ongrádi"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},chapters:[{id:"51751",title:"Introductory Chapter: Unconventional Philosophy of Herpesvirus Infections",slug:"introductory-chapter-unconventional-philosophy-of-herpesvirus-infections",totalDownloads:637,totalCrossrefCites:0,signatures:"Joseph Ongrádi",authors:[{id:"36151",title:"Dr.",name:"Joseph",middleName:null,surname:"Ongrádi",fullName:"Joseph Ongrádi",slug:"joseph-ongradi"}]},{id:"51604",title:"Herpes Simplex Virus Type 1 at the Central Nervous System",slug:"herpes-simplex-virus-type-1-at-the-central-nervous-system",totalDownloads:1043,totalCrossrefCites:2,signatures:"Carola Otth, Francisca Acuña‐Hinrichsen, Luis Leyton, Carolina\nMartin and Margarita I. Concha",authors:[{id:"184195",title:"Ph.D.",name:"Carola",middleName:null,surname:"Otth",fullName:"Carola Otth",slug:"carola-otth"},{id:"184196",title:"MSc.",name:"Francisca",middleName:null,surname:"Acuña-Hinrichsen",fullName:"Francisca Acuña-Hinrichsen",slug:"francisca-acuna-hinrichsen"},{id:"184197",title:"BSc.",name:"Luis",middleName:null,surname:"Leyton",fullName:"Luis Leyton",slug:"luis-leyton"},{id:"184198",title:"Dr.",name:"Margarita",middleName:null,surname:"Concha",fullName:"Margarita Concha",slug:"margarita-concha"},{id:"187592",title:"Dr.",name:"Carolina",middleName:null,surname:"Martin",fullName:"Carolina Martin",slug:"carolina-martin"}]},{id:"51338",title:"Herpesviruses in Periapical Pathoses: An Updated Systematic Review",slug:"herpesviruses-in-periapical-pathoses-an-updated-systematic-review",totalDownloads:733,totalCrossrefCites:1,signatures:"Aleksandar Jakovljević, Miroslav Andrić, Aleksandra Knežević,\nKatarina Beljić-Ivanović, Maja Miletić, Tanja Jovanović, Ljiljana Kesić\nand Jelena Milašin",authors:[{id:"49081",title:"Dr.",name:"Miroslav",middleName:null,surname:"Andric",fullName:"Miroslav Andric",slug:"miroslav-andric"},{id:"64273",title:"Dr.",name:"Jelena",middleName:null,surname:"Milašin",fullName:"Jelena Milašin",slug:"jelena-milasin"},{id:"183895",title:"Dr.",name:"Aleksandar",middleName:null,surname:"Jakovljevic",fullName:"Aleksandar Jakovljevic",slug:"aleksandar-jakovljevic"},{id:"184099",title:"Prof.",name:"Aleksandra",middleName:null,surname:"Knezevic",fullName:"Aleksandra Knezevic",slug:"aleksandra-knezevic"},{id:"184100",title:"Dr.",name:"Katarina",middleName:null,surname:"Beljic-Ivanovic",fullName:"Katarina Beljic-Ivanovic",slug:"katarina-beljic-ivanovic"},{id:"184101",title:"Dr.",name:"Maja",middleName:null,surname:"Miletic",fullName:"Maja Miletic",slug:"maja-miletic"},{id:"184104",title:"Prof.",name:"Tanja",middleName:null,surname:"Jovanovic",fullName:"Tanja Jovanovic",slug:"tanja-jovanovic"},{id:"184105",title:"Prof.",name:"Ljiljana",middleName:null,surname:"Kesic",fullName:"Ljiljana Kesic",slug:"ljiljana-kesic"}]},{id:"51491",title:"Ectopic Expression of Human Herpesvirus 1 Thymidine Kinase Induces Male Infertility",slug:"ectopic-expression-of-human-herpesvirus-1-thymidine-kinase-induces-male-infertility",totalDownloads:422,totalCrossrefCites:0,signatures:"Mo Chen, Li‐yi Cai, Takako Kato and Yukio Kato",authors:[{id:"99373",title:"Prof.",name:"Yukio",middleName:null,surname:"Kato",fullName:"Yukio Kato",slug:"yukio-kato"},{id:"99403",title:"Dr.",name:"Takako",middleName:null,surname:"Kato",fullName:"Takako Kato",slug:"takako-kato"},{id:"188605",title:"Dr.",name:"Mo",middleName:null,surname:"Chen",fullName:"Mo Chen",slug:"mo-chen"},{id:"188606",title:"Dr.",name:"Li-Yi",middleName:null,surname:"Cai",fullName:"Li-Yi Cai",slug:"li-yi-cai"}]},{id:"51597",title:"Immune Evasion by Herpes Simplex Viruses",slug:"immune-evasion-by-herpes-simplex-viruses",totalDownloads:758,totalCrossrefCites:1,signatures:"Angello R. Retamal-Díaz, Eduardo Tognarelli, Alexis M. Kalergis,\nSusan M. Bueno and Pablo A. González",authors:[{id:"182617",title:"Dr.",name:"Pablo",middleName:"Alberto",surname:"Gonzalez",fullName:"Pablo Gonzalez",slug:"pablo-gonzalez"},{id:"183529",title:"BSc.",name:"Eduardo",middleName:null,surname:"Tognarelli",fullName:"Eduardo Tognarelli",slug:"eduardo-tognarelli"},{id:"183530",title:"MSc.",name:"Angello",middleName:null,surname:"Retamal",fullName:"Angello Retamal",slug:"angello-retamal"},{id:"187559",title:"Dr.",name:"Susan",middleName:null,surname:"Bueno",fullName:"Susan Bueno",slug:"susan-bueno"},{id:"187560",title:"Dr.",name:"Alexis",middleName:null,surname:"Kalergis",fullName:"Alexis Kalergis",slug:"alexis-kalergis"}]},{id:"51911",title:"Review: Biological and Pharmacological Basis of Cytolytic Viral Activation in EBV-Associated Nasopharyngeal Carcinoma",slug:"review-biological-and-pharmacological-basis-of-cytolytic-viral-activation-in-ebv-associated-nasophar",totalDownloads:1081,totalCrossrefCites:0,signatures:"Natalie Oker, Nikiforos-Ioannis Kapetanakis and Pierre Busson",authors:[{id:"183209",title:"D.Sc.",name:"Pierre",middleName:null,surname:"Busson",fullName:"Pierre Busson",slug:"pierre-busson"}]},{id:"51617",title:"Herpes Simplex Virus 1 and 2 Vaccine Design: What can we Learn from the Past?",slug:"herpes-simplex-virus-1-and-2-vaccine-design-what-can-we-learn-from-the-past-",totalDownloads:917,totalCrossrefCites:2,signatures:"Vladimíra Ďurmanová, Marian Adamkov and Július Rajčáni",authors:[{id:"181773",title:"Emeritus Prof.",name:"Julius",middleName:null,surname:"Rajčáni",fullName:"Julius Rajčáni",slug:"julius-rajcani"},{id:"185924",title:"Prof.",name:"Marian",middleName:null,surname:"Adamkov",fullName:"Marian Adamkov",slug:"marian-adamkov"},{id:"186238",title:"Associate Prof.",name:"Vladimira",middleName:null,surname:"Ďurmanová",fullName:"Vladimira Ďurmanová",slug:"vladimira-durmanova"}]},{id:"50493",title:"Herpesvirus in Bovines: Importance of Bovine Herpesvirus Type 1",slug:"herpesvirus-in-bovines-importance-of-bovine-herpesvirus-type-1",totalDownloads:1023,totalCrossrefCites:0,signatures:"Gurpreet Kaur and Mudit Chandra",authors:[{id:"183506",title:"Dr.",name:"Gurpreet",middleName:null,surname:"Kaur",fullName:"Gurpreet Kaur",slug:"gurpreet-kaur"},{id:"185854",title:"Dr.",name:"Mudit",middleName:null,surname:"Chandra",fullName:"Mudit Chandra",slug:"mudit-chandra"}]},{id:"51229",title:"Latency of Bovine Herpesvirus 1 (BoHV-1) in Sensory Neurons",slug:"latency-of-bovine-herpesvirus-1-bohv-1-in-sensory-neurons",totalDownloads:671,totalCrossrefCites:1,signatures:"Clinton Jones",authors:[{id:"183920",title:"Ph.D.",name:"Clinton",middleName:null,surname:"Jones",fullName:"Clinton Jones",slug:"clinton-jones"}]}]},relatedBooks:[{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"},chapters:[{id:"41407",title:"The Phylogeny and Classification of Anopheles",slug:"the-phylogeny-and-classification-of-anopheles",signatures:"Ralph E. Harbach",authors:[{id:"151606",title:"Dr.",name:"Ralph",middleName:null,surname:"E. Harbach",fullName:"Ralph E. Harbach",slug:"ralph-e.-harbach"}]},{id:"43979",title:"Systematic Techniques for the Recognition of Anopheles Species Complexes",slug:"systematic-techniques-for-the-recognition-of-anopheles-species-complexes",signatures:"Wej Choochote and Atiporn Saeung",authors:[{id:"151262",title:"Prof.",name:"Wej",middleName:null,surname:"Choochote",fullName:"Wej Choochote",slug:"wej-choochote"},{id:"153513",title:"Dr.",name:"Atiporn",middleName:null,surname:"Saeung",fullName:"Atiporn Saeung",slug:"atiporn-saeung"}]},{id:"43884",title:"Genetic and Phenetic Approaches to Anopheles Systematics",slug:"genetic-and-phenetic-approaches-to-anopheles-systematics",signatures:"Claire Garros and Jean-Pierre Dujardin",authors:[{id:"107247",title:"Dr.",name:"Jean-Pierre",middleName:null,surname:"Dujardin",fullName:"Jean-Pierre Dujardin",slug:"jean-pierre-dujardin"},{id:"151616",title:"Dr.",name:"Claire",middleName:null,surname:"Garros",fullName:"Claire Garros",slug:"claire-garros"}]},{id:"43624",title:"Global Distribution of the Dominant Vector Species of Malaria",slug:"global-distribution-of-the-dominant-vector-species-of-malaria",signatures:"Marianne E. Sinka",authors:[{id:"153626",title:"Dr.",name:"Marianne",middleName:null,surname:"Sinka",fullName:"Marianne Sinka",slug:"marianne-sinka"}]},{id:"43482",title:"Phylogeography, Vectors and Transmission in Latin America",slug:"phylogeography-vectors-and-transmission-in-latin-america",signatures:"Jan E. Conn, Martha L. Quiñones and Marinete M. Póvoa",authors:[{id:"151253",title:"Prof.",name:"Jan",middleName:null,surname:"Conn",fullName:"Jan Conn",slug:"jan-conn"},{id:"154130",title:"Prof.",name:"Martha",middleName:null,surname:"Quinones",fullName:"Martha Quinones",slug:"martha-quinones"},{id:"154131",title:"Prof.",name:"Marinete",middleName:null,surname:"Povoa",fullName:"Marinete Povoa",slug:"marinete-povoa"}]},{id:"44284",title:"Speciation in Anopheles gambiae — The Distribution of Genetic Polymorphism and Patterns of Reproductive Isolation Among Natural Populations",slug:"speciation-in-anopheles-gambiae-the-distribution-of-genetic-polymorphism-and-patterns-of-reproductiv",signatures:"Gregory C. Lanzaro and Yoosook Lee",authors:[{id:"152068",title:"Prof.",name:"Gregory C.",middleName:null,surname:"Lanzaro",fullName:"Gregory C. Lanzaro",slug:"gregory-c.-lanzaro"},{id:"169011",title:"Dr.",name:"Yoosook",middleName:null,surname:"Lee",fullName:"Yoosook Lee",slug:"yoosook-lee"}]},{id:"43973",title:"Advances and Perspectives in the Study of the Malaria Mosquito Anopheles funestus",slug:"advances-and-perspectives-in-the-study-of-the-malaria-mosquito-anopheles-funestus",signatures:"Ibrahima Dia, Moussa Wamdaogo Guelbeogo and Diego Ayala",authors:[{id:"154416",title:"Dr.",name:"Diego",middleName:null,surname:"Ayala",fullName:"Diego Ayala",slug:"diego-ayala"},{id:"167122",title:"Dr.",name:"Ibrahima",middleName:null,surname:"Dia",fullName:"Ibrahima Dia",slug:"ibrahima-dia"},{id:"169020",title:"Dr.",name:"Moussa",middleName:"Wamdaogo",surname:"Guelbeogo",fullName:"Moussa Guelbeogo",slug:"moussa-guelbeogo"}]},{id:"43614",title:"Highlights on Anopheles nili and Anopheles moucheti, Malaria Vectors in Africa",slug:"highlights-on-anopheles-nili-and-anopheles-moucheti-malaria-vectors-in-africa",signatures:"Christophe Antonio-Nkondjio and Frédéric Simard",authors:[{id:"153999",title:"Dr.",name:"Christophe",middleName:null,surname:"Antonio Nkondjio",fullName:"Christophe Antonio Nkondjio",slug:"christophe-antonio-nkondjio"},{id:"154272",title:"Dr.",name:"Frédéric",middleName:null,surname:"Simard",fullName:"Frédéric Simard",slug:"frederic-simard"}]},{id:"43975",title:"The Dominant Mosquito Vectors of Human Malaria in India",slug:"the-dominant-mosquito-vectors-of-human-malaria-in-india",signatures:"Vas Dev and Vinod P. Sharma",authors:[{id:"151166",title:"Dr.",name:"Vas",middleName:null,surname:"Dev",fullName:"Vas Dev",slug:"vas-dev"},{id:"169007",title:"Dr.",name:"Vinod",middleName:null,surname:"P. Sharma",fullName:"Vinod P. Sharma",slug:"vinod-p.-sharma"}]},{id:"45385",title:"Vector Biology and Malaria Transmission in Southeast Asia",slug:"vector-biology-and-malaria-transmission-in-southeast-asia",signatures:"Wannapa Suwonkerd, Wanapa Ritthison, Chung Thuy Ngo, Krajana\nTainchum, Michael J. Bangs and Theeraphap Chareonviriyaphap",authors:[{id:"151663",title:"PhD.",name:"Wannapa",middleName:null,surname:"Suwonkerd",fullName:"Wannapa Suwonkerd",slug:"wannapa-suwonkerd"},{id:"151737",title:"Dr.",name:"Michael",middleName:null,surname:"J. Bangs",fullName:"Michael J. Bangs",slug:"michael-j.-bangs"},{id:"169010",title:"Dr.",name:"Wanapa",middleName:null,surname:"Ritthison",fullName:"Wanapa Ritthison",slug:"wanapa-ritthison"}]},{id:"43254",title:"Understanding Anopheles Diversity in Southeast Asia and Its Applications for Malaria Control",slug:"understanding-anopheles-diversity-in-southeast-asia-and-its-applications-for-malaria-control",signatures:"Katy Morgan, Pradya Somboon and Catherine Walton",authors:[{id:"154092",title:"Dr.",name:"Catherine",middleName:null,surname:"Walton",fullName:"Catherine Walton",slug:"catherine-walton"},{id:"154867",title:"Dr.",name:"Katy",middleName:null,surname:"Morgan",fullName:"Katy Morgan",slug:"katy-morgan"},{id:"169019",title:"Dr.",name:"Pradya",middleName:null,surname:"Somboon",fullName:"Pradya Somboon",slug:"pradya-somboon"}]},{id:"44155",title:"The Systematics and Bionomics of Malaria Vectors in the Southwest Pacific",slug:"the-systematics-and-bionomics-of-malaria-vectors-in-the-southwest-pacific",signatures:"Nigel W. Beebe, Tanya L. Russell, Thomas R. Burkot, Neil F. Lobo and\nRobert D. Cooper",authors:[{id:"152080",title:"Dr.",name:"Nigel",middleName:null,surname:"Beebe",fullName:"Nigel Beebe",slug:"nigel-beebe"},{id:"169012",title:"Dr.",name:"Tanya",middleName:null,surname:"L. Russell",fullName:"Tanya L. Russell",slug:"tanya-l.-russell"},{id:"169013",title:"Dr.",name:"Thomas",middleName:null,surname:"R. Burkot",fullName:"Thomas R. Burkot",slug:"thomas-r.-burkot"},{id:"169014",title:"Dr.",name:"Neil",middleName:null,surname:"F. Lobo",fullName:"Neil F. Lobo",slug:"neil-f.-lobo"},{id:"169015",title:"Dr.",name:"Robert",middleName:null,surname:"D. Cooper",fullName:"Robert D. Cooper",slug:"robert-d.-cooper"}]},{id:"43671",title:"Ecology of Larval Habitats",slug:"ecology-of-larval-habitats",signatures:"Eliška Rejmánková, John Grieco, Nicole Achee and Donald R.\nRoberts",authors:[{id:"151632",title:"Prof.",name:"Nicole",middleName:null,surname:"Achee",fullName:"Nicole Achee",slug:"nicole-achee"},{id:"152601",title:"Prof.",name:"Eliska",middleName:null,surname:"Rejmankova",fullName:"Eliska Rejmankova",slug:"eliska-rejmankova"},{id:"169016",title:"Dr.",name:"John",middleName:null,surname:"Grieco",fullName:"John Grieco",slug:"john-grieco"}]},{id:"43954",title:"From Anopheles to Spatial Surveillance: A Roadmap Through a Multidisciplinary Challenge",slug:"from-anopheles-to-spatial-surveillance-a-roadmap-through-a-multidisciplinary-challenge",signatures:"Valérie Obsomer, Nicolas Titeux, Christelle Vancustem, Grégory\nDuveiller, Jean-François Pekel, Steve Connor, Pietro Ceccato and\nMarc Coosemans",authors:[{id:"131417",title:"Dr.",name:"Valérie",middleName:null,surname:"Obsomer",fullName:"Valérie Obsomer",slug:"valerie-obsomer"},{id:"152754",title:"Prof.",name:"Marc",middleName:null,surname:"Coosemans",fullName:"Marc Coosemans",slug:"marc-coosemans"},{id:"153949",title:"Dr.",name:"Pietro",middleName:null,surname:"Ceccato",fullName:"Pietro Ceccato",slug:"pietro-ceccato"},{id:"153950",title:"Dr.",name:"Gregory",middleName:null,surname:"Duveiller",fullName:"Gregory Duveiller",slug:"gregory-duveiller"},{id:"153952",title:"Dr.",name:"Christelle",middleName:null,surname:"Vancutsem",fullName:"Christelle Vancutsem",slug:"christelle-vancutsem"},{id:"153980",title:"Dr.",name:"Nicolas",middleName:null,surname:"Titeux",fullName:"Nicolas Titeux",slug:"nicolas-titeux"},{id:"154158",title:"Dr.",name:"Steve J",middleName:null,surname:"Connor",fullName:"Steve J Connor",slug:"steve-j-connor"},{id:"167685",title:"MSc.",name:"Jean-Francois",middleName:null,surname:"Pekel",fullName:"Jean-Francois Pekel",slug:"jean-francois-pekel"}]},{id:"43960",title:"Simian Malaria Parasites: Special Emphasis on Plasmodium knowlesi and Their Anopheles Vectors in Southeast Asia",slug:"simian-malaria-parasites-special-emphasis-on-plasmodium-knowlesi-and-their-anopheles-vectors-in-sout",signatures:"Indra Vythilingam and Jeffery Hii",authors:[{id:"151116",title:"Dr.",name:"Indra",middleName:null,surname:"Vythilingam",fullName:"Indra Vythilingam",slug:"indra-vythilingam"},{id:"169006",title:"Dr.",name:"Jeffery",middleName:null,surname:"Hii",fullName:"Jeffery Hii",slug:"jeffery-hii"}]},{id:"44039",title:"Thermal Stress and Thermoregulation During Feeding in Mosquitoes",slug:"thermal-stress-and-thermoregulation-during-feeding-in-mosquitoes",signatures:"Chloé Lahondère and Claudio R. Lazzari",authors:[{id:"151619",title:"Prof.",name:"Claudio",middleName:null,surname:"R. Lazzari",fullName:"Claudio R. Lazzari",slug:"claudio-r.-lazzari"},{id:"151620",title:"Ms.",name:"Chloé",middleName:null,surname:"Lahondère",fullName:"Chloé Lahondère",slug:"chloe-lahondere"}]},{id:"43955",title:"The Anopheles Mosquito Microbiota and Their Impact on Pathogen Transmission",slug:"the-anopheles-mosquito-microbiota-and-their-impact-on-pathogen-transmission",signatures:"Mathilde Gendrin and George K. Christophides",authors:[{id:"154007",title:"Dr.",name:"Mathilde",middleName:null,surname:"Gendrin",fullName:"Mathilde Gendrin",slug:"mathilde-gendrin"},{id:"154008",title:"Prof.",name:"George",middleName:"K",surname:"Christophides",fullName:"George Christophides",slug:"george-christophides"}]},{id:"43829",title:"Bacterial Biodiversity in Midguts of Anopheles Mosquitoes, Malaria Vectors in Southeast Asia",slug:"bacterial-biodiversity-in-midguts-of-anopheles-mosquitoes-malaria-vectors-in-southeast-asia",signatures:"Sylvie Manguin, Chung Thuy Ngo, Krajana Tainchum, Waraporn\nJuntarajumnong, Theeraphap Chareonviriyaphap, Anne-Laure\nMichon and Estelle Jumas-Bilak",authors:[{id:"50017",title:"Prof.",name:"Sylvie",middleName:null,surname:"Manguin",fullName:"Sylvie Manguin",slug:"sylvie-manguin"},{id:"75315",title:"Prof.",name:"Theeraphap",middleName:null,surname:"Chareonviriyaphap",fullName:"Theeraphap Chareonviriyaphap",slug:"theeraphap-chareonviriyaphap"},{id:"88985",title:"Prof.",name:"Anne-Laure",middleName:null,surname:"Michon",fullName:"Anne-Laure Michon",slug:"anne-laure-michon"},{id:"88986",title:"Prof.",name:"Estelle",middleName:null,surname:"Jumas-Bilak",fullName:"Estelle Jumas-Bilak",slug:"estelle-jumas-bilak"},{id:"156016",title:"MSc.",name:"Chung Thuy",middleName:null,surname:"Ngo",fullName:"Chung Thuy Ngo",slug:"chung-thuy-ngo"},{id:"156018",title:"MSc.",name:"Krajana",middleName:null,surname:"Tainchum",fullName:"Krajana Tainchum",slug:"krajana-tainchum"},{id:"156019",title:"Dr.",name:"Waraporn",middleName:null,surname:"Juntarajumnong",fullName:"Waraporn Juntarajumnong",slug:"waraporn-juntarajumnong"}]},{id:"43899",title:"Distribution, Mechanisms, Impact and Management of Insecticide Resistance in Malaria Vectors: A Pragmatic Review",slug:"distribution-mechanisms-impact-and-management-of-insecticide-resistance-in-malaria-vectors-a-pragmat",signatures:"Vincent Corbel and Raphael N’Guessan",authors:[{id:"152666",title:"Dr.",name:"Vincent",middleName:null,surname:"Corbel",fullName:"Vincent Corbel",slug:"vincent-corbel"},{id:"169017",title:"Dr.",name:"Raphael",middleName:null,surname:"N'Guessan",fullName:"Raphael N'Guessan",slug:"raphael-n'guessan"}]},{id:"43851",title:"Perspectives on Barriers to Control of Anopheles Mosquitoes and Malaria",slug:"perspectives-on-barriers-to-control-of-anopheles-mosquitoes-and-malaria",signatures:"Donald R. Roberts, Richard Tren and Kimberly Hess",authors:[{id:"151439",title:"Prof.",name:"Donald",middleName:null,surname:"R. Roberts",fullName:"Donald R. Roberts",slug:"donald-r.-roberts"},{id:"151656",title:"Mr.",name:"Richard",middleName:null,surname:"Tren",fullName:"Richard Tren",slug:"richard-tren"},{id:"154152",title:"Ms.",name:"Kimberly",middleName:null,surname:"Hess",fullName:"Kimberly Hess",slug:"kimberly-hess"}]},{id:"43874",title:"Residual Transmission of Malaria: An Old Issue for New Approaches",slug:"residual-transmission-of-malaria-an-old-issue-for-new-approaches",signatures:"Lies Durnez and Marc Coosemans",authors:[{id:"152754",title:"Prof.",name:"Marc",middleName:null,surname:"Coosemans",fullName:"Marc Coosemans",slug:"marc-coosemans"},{id:"169018",title:"Dr.",name:"Lies",middleName:null,surname:"Durnez",fullName:"Lies Durnez",slug:"lies-durnez"}]},{id:"44330",title:"Vector Control: Some New Paradigms and Approaches",slug:"vector-control-some-new-paradigms-and-approaches",signatures:"Claire Duchet, Richard Allan and Pierre Carnevale",authors:[{id:"151662",title:"Dr.",name:"Pierre",middleName:null,surname:"Carnevale",fullName:"Pierre Carnevale",slug:"pierre-carnevale"},{id:"169000",title:"Dr.",name:"Richard",middleName:null,surname:"Allan",fullName:"Richard Allan",slug:"richard-allan"},{id:"169008",title:"Dr.",name:"Claire",middleName:null,surname:"Duchet",fullName:"Claire Duchet",slug:"claire-duchet"}]},{id:"43870",title:"New Salivary Biomarkers of Human Exposure to Malaria Vector Bites",slug:"new-salivary-biomarkers-of-human-exposure-to-malaria-vector-bites",signatures:"Papa M. Drame, Anne Poinsignon, Alexandra Marie, Herbert\nNoukpo, Souleymane Doucoure, Sylvie Cornelie and Franck\nRemoue",authors:[{id:"151515",title:"Dr.",name:"Papa Makhtar",middleName:null,surname:"Drame",fullName:"Papa Makhtar Drame",slug:"papa-makhtar-drame"},{id:"151648",title:"Dr.",name:"Franck",middleName:null,surname:"Remoué",fullName:"Franck Remoué",slug:"franck-remoue"},{id:"154034",title:"Dr.",name:"Anne",middleName:null,surname:"Poinsignon",fullName:"Anne Poinsignon",slug:"anne-poinsignon"},{id:"154035",title:"MSc.",name:"Alexandra",middleName:null,surname:"Marie",fullName:"Alexandra Marie",slug:"alexandra-marie"},{id:"154037",title:"Dr.",name:"Souleymane",middleName:null,surname:"Doucoure",fullName:"Souleymane Doucoure",slug:"souleymane-doucoure"},{id:"154038",title:"MSc.",name:"Herbert",middleName:null,surname:"Noukpo",fullName:"Herbert Noukpo",slug:"herbert-noukpo"},{id:"154039",title:"Dr.",name:"Sylvie",middleName:null,surname:"Cornélie",fullName:"Sylvie Cornélie",slug:"sylvie-cornelie"}]},{id:"44149",title:"Transgenic Mosquitoes for Malaria Control: From the Bench to the Public Opinion Survey",slug:"transgenic-mosquitoes-for-malaria-control-from-the-bench-to-the-public-opinion-survey",signatures:"Christophe Boëte and Uli Beisel",authors:[{id:"98400",title:"Dr.",name:"Christophe",middleName:null,surname:"Boëte",fullName:"Christophe Boëte",slug:"christophe-boete"},{id:"167749",title:"Dr.",name:"Uli",middleName:null,surname:"Beisel",fullName:"Uli Beisel",slug:"uli-beisel"}]}]}]},onlineFirst:{chapter:{type:"chapter",id:"67937",title:"Effect of Essential Oils on Storability and Preservation of Some Vegetable Crops",doi:"10.5772/intechopen.87213",slug:"effect-of-essential-oils-on-storability-and-preservation-of-some-vegetable-crops",body:'
1. Introduction
Recently, the natural alternatives such as plant essential oils provide a promising control of plant diseases and anti-sprout agent because they virtually constitute a rich source of bioactive chemicals such as phenols, flavonoids, quinones, tannins, alkaloids, saponins, sterols terpenes, aromatic and aldehydes [1]. Moreover, these natural alternatives can also maintain the biochemical constituents of tubers during storage, they are biodegradable to nontoxic products, and are potentially suitable for use in integrated pest management programs.
Jerusalem artichoke JA or sun choke (Helianthus tuberosus L.) is a perennial plant which has a high economic value. JA used traditionally for human food and livestock feed due to its high nutritive value. JA tubers used for production of biofuels (ethanol) and some functional food like inulin, fructooligosaccharides and fructose. Moreover, some bioactive metabolites from its leaves and stems have been used in some pharmaceutical industries [1]. Storage JA tuber, controlled rots can be done by various techniques including; cold temperature, removal of diseases in tubers and minimizing mechanical injuries or application of synthetic fungicides. Another simple applied method in developing countries is keeping the tubers in the soil. Unfortunately, many fungi diseases can grow at cold storage temperatures and lead to damage, especially in extending long storage [2], However, storage of the harvested tubers usually results in high losses in quality, caused mainly by desiccation, rotting, sprouting, freezing and inulin degradation. A common solution is the use of synthetic chemical fungicides, however, their use is accompanied by threatening human health and the environment by supporting the emergence of resistant pathogens and by contamination of food with the pesticide deposits [3]. Essential oils, as green fungicides, are emerging as a better alternative of synthetic fungicides due to their high efficacy, biodegradability, eco-safety and volatile nature.
Respiration of potato tubers during storage and breakdown of dormancy during storage result in sprouting and loss of nutritive value of tubers [4]. Sprouting reduces the weight, the nutritional and processing quality of tubers and the number of marketable potatoes, being responsible for important economic losses during potatoes storage [5]. These physiological changes affect the internal composition of the tuber and destruction of edible material and changes in nutritional quality [6]. Various methods are available to control sprouting during storage. The primary method to control sprouting in storage is with postharvest application of isopropyl N-(3-chlorophenyl) carbamate (chlorpropham; CIPC). CIPC inhibits sprout development by interfering with cell division [7]. Therefore, a pressing need exists to find other, more environmentally acceptable sprout inhibitors for tubers. Nowadays it’s very important to use natural products compounds such as essential oils.
Broccoli sprouts are considered as a functional food. Essential nutrient content provides diverse secondary metabolites and phytochemicals [8]. The phenolic compounds, especially flavonoids and anthocyanin, show a great ability capture free radical that leading to oxidative stress, to these compounds are attributed a beneficial effect in the prevention of cardiovascular diseases, circulatory problems, neurological disorders and cancer [9]. Broccoli has been identified as a vegetable with potential anti-cancer activity due to high levels of glucosinolates. The use of essential oil treatments rich in antioxidant to stimulate broccoli seed germination should be considered. Application of thyme and basil oil at 4% reduced the pathogenic fungi from seed to seedling and had a positive effect on the seed germination of infected seeds [10]. Aromatic plants especially essential oils are well known for their antioxidant and antimicrobial properties that prevent food degradation and alteration [11], as they are rich in phenolic substances, usually referred to as polyphenols, which are ubiquitous components of plants and herbs.
2. Application of essential oils
2.1 Alternative preservation method against sclerotium tuber rot of Jerusalem artichoke using natural essential oils
2.1.1 Methodology
Two experiments were conducted in Mansoura laboratory for vegetable crop handling and postharvest according to the storage method. In the first experiment, the tubers were kept in perforated polyethylene bags (0.075 mm thickness), and stored at 4°C and 90–95% relative humidity RH. In the second experiment, the tubers were stored in carton boxes (3 m3) at 25 ± 2°C with moistened peat moss layers at the rate of peat moss: JA tubers (1.5: 1, kg/kg). The treatments applied for each experiment can be summarized as follows: Control (C), infected with fungal pathogen S. rolfsii (P), treated with caraway essential oil (O) and treated with caraway oil and infected with pathogen S. rolfsii (O + P). About 30 kg of tubers was used for each treatment.
2.1.2 Important results
2.1.2.1 Antifungal activity of the essential oils
Assessment of antifungal activity in vitro of caraway and spearmint essential oils was evaluated against S. rolfsii (Figure 1). Caraway essential oil completely inhibited the growth of the fungal pathogen even at the lowest concentration (2%). On the other hand, spearmint essential oil showed slight reduction in the fungal pathogen growth. The antifungal activity of caraway essential oil may be attributed to some antifungal phytochemicals that constitute a large fraction of the oil like carvone, limonene, carveol, pinene and thujone [12].
Figure 1.
Inhibition in mycelial dry weight of S. rolfsii as a function of the tested oils. Columns superscripted with the same letter are not significantly different at P ≤ 0.05 (Duncan test).
2.1.2.2 Evaluation of the caraway essential oil and peat moss application under storage conditions
2.1.2.2.1 Severity of disease
Data presented in Table 1 show the rot fugal disease severity of JA tubers exposed to caraway oil and infected with fungal pathogen S. rolfsii under the two storage methods. The disease severity increased with the increasing the storage period over to the storage methods. JA tubers infected with S. rolfsii and exposed to emulsion of caraway essential oil (O + P) in peat moss layer at 25°C significantly reduced the disease severity compared to the cold storage method after 4 months storage. Infected-control JA tubers (P) and storage in peat moss layer at 25°C significantly reduced the disease severity for 2 months of storage compared with cold storage method, after which, the tubers were fully decadent. On the other hand, control-uninfected JA tubers (C) and storage in peat moss layer at 25°C significantly reduced the disease severity (caused by reasons other than S. rolfsii) compared with the storage under cold storage method. Caraway essential oil had the antimicrobial effects due to its content of basic constituents of monoterpenes, carvone and limonene. The basic constituents had a permeability effect on fungal cell membrane, inactivation of some organic compounds and enzymes and/or the inhibition of conidial germination, eventually, the death of fungal pathogen [13]. Moreover, the use of essential oils in storage of JA may have many benefits, including, they are natural-ecofriendly products, biodegradable and multifunctional purposes.
Mean rot severity (%) of JA tubers treated with caraway essential oil and infected with S. rolfsii using two storage methods.
1 = storage JA tubers in polyethylene bags at 4°C and 2 = storage JA tubers in peat moss layer at 25°C.
C = untreated control, P = infected tubers with pathogen, O = treated JA tubers with caraway essential oil and O + P = infected tubers with pathogen and treated with caraway essential oil. NA = not applicable due to full decay.
Means in each column followed by the same letter(s) were not significant at P= 0.05; where, (a) refer to the highest mean values, and (e) refer to the lowest mean values according to Duncan multiple range test.
Tubers rot in the untreated control (C) treatment was due to many reasons rather than the fungal pathogens.
Moreover, the activity of essential oils against may tend to reduce pathogenic fungi resistance reinforcement against chemical fungicide because they contain two or more stereo-isomers that may be located on multi-sites on the pathogen’s plasma membrane. One of the valuable applications for peat moss is the traditional use in food preservation [14]. The antifungal effect of the use of peat mosses has been reported by many investigators against Aspergillus niger, A. flavus, Candida albicans, Cryptococcus albidus and Trichophyton rubrum [15]. The antifungal effect of peat moss may be related to some of its contains of extranutritional constituents or bioactive components like a pectin-like polymer and sphagnan, that inhibit fungal mycelium growth via electrostatic immobilization of extracellular enzymes and/or nitrogen deprivation, phenolics that inhibit the activity of extracellular enzymes of microbes or other constituents like sterols and polyacetylenes [16].
2.1.2.2.2 Sprouting, weight loss and dry matter percentages of JA tubers
Table 2 show the mean data of weight loss and dry matter percentages of JA tubers exposed to emulsion of caraway essential oil and infected with fungal pathogenic S. rolfsii under using the two different storage methods. Results indicated that, the treatment of healthy JA tubers with emulsion of caraway essential oil completely inhibited the tubers sprouting and weight loss, but recorded the highest dry mater weight percentage along the storage period compared with the untreated-uninfected control treatment over the use of the two different methods. Even after 120 days of storage period, the treatment of the use of JA tubers with caraway oil and infected with pathogenic fungi significantly decreased sprouting and weight loss percentages and increased dry matter content for JA tubers that stored in peat moss layers at 25°C than those stored in polyethylene bags at 4°C when compared with the control (infected-untreated) tubers. On the other hand, storage of the untreated-uninfected JA tubers in peat moss layers at 25°C increased the sprouting, and dry matter content and decreased the weight of the tubers compared to the storage of tubers in polyethylene bags at 4°C. The bioactive components like limonene and carvone, in caraway essential oil are known to inhabit sprouting percentage of JA tubers by the suppressing of mitochondrial respiration and reducing carbohydrate deterioration sugar content. Carvone had a specific tool for inhibition of sprout growth of potato tubers, such as the repression of key enzyme in the mevalonate acid pathway, which is the main precursor of gibberellin biosynthesis [17]. On the other hand, peat moss has a relatively high water retention capacity; their cells can hold 16–25 times their dry weight of water [14]. This in turns encourages such amendment for its use in the preservation of JA tubers by increasing a relative humidity around the tubers and preventing heat transfer within the peat moss layer leading to the decrease of the water loss from fresh tubers depends on the difference between the water vapor pressure within the tubers and the water vapor pressure of the surrounding air, with moisture passing from the higher pressure to the lower even at 25°C. Cabezas et al. [18] reported that dry matter content in JA tubers decreased significantly depends on many factors, such as storage conditions, storage periods and keeping tubers for 30 days at 18°C, this leads to loosing water above 20%.
Mean sprouting, weight loss and dry matter weight (% ± 2SD) of JA tubers treated with caraway essential oil and infected with S. rolfsii using two storage methods.
1 = storage JA tubers in polyethylene bags at 4°C and 2 = storage JA tubers in peat moss layer at 25°C.
C = untreated control, P = infected tubers with pathogen, O = treated JA tubers with caraway essential oil and O + P = infected tubers with pathogen and treated with caraway essential oil.
NA = not applicable due to full decay.
2.1.2.2.3 Biochemical constituents of JA tubers
Table 3 show the data of carbohydrates content, inulin and protein in JA tubers exposed to emulsion of caraway essential oil and then infected with fungal pathogenic S. rolfsii over the use of two storage methods. The application of caraway essential oil and uninfected JA tubers had significant effects on total carbohydrates, inulin and protein contents compared with the untreated-uninfected control in both storage methods. Along 4 months of storage, the treatment of infected JA tubers with pathogen and treated with caraway essential oil effectively decreased the carbohydrate, inulin and protein contents compared with the infected-untreated control JA tubers in both methods of storage. A fresh JA tuber contains 80% water, 15% carbohydrates, mainly in the form of inulin and about 2% protein in dry matter [19]. There are many changes in fresh JA tubers with long term storage, i.e., physical, biochemical, microbiological and enzymatic and which may lead to tuber decay. To inhibit these biochemical activities, natural or artificial drying products are widely used [20]. Davies [21] reported that the basic constituents of caraway oil (monoterpenes) tend to delay and the deterioration of carbohydrates and protein contents associated with the enzymatic system as well as respiration and energy metabolism enzyme keeping the internal biochemical enzymatic activities in minimum level.
Mean contents of carbohydrates, inulin (mg/g ± 2SD) and protein (% ± 2SD) of JA tubers treated with caraway essential oil and infected with S. rolfsii using two storage methods.
1 = storage JA tubers in polyethylene bags at 4°C and 2 = storage JA tubers in peat moss layer at 25°C.
C = untreated control, P = infected tubers with pathogen, O = treated JA tubers with caraway essential oil and O + P = infected tubers with pathogen and treated with caraway essential oil.
NA = not applicable due to full decay.
2.1.2.2.4 Peroxidase, polyphenoloxidase enzymes and phenol content in JA tubers
The mean activities data of peroxidase, polyphenoloxidase and phenol contents of JA fresh tubers treated with caraway essential oils and infected with pathogenic fungi over the use of two different two storage methods are presented on Table 4. Results revealed that infection with S. rolfsii had significant effects on total phenol and the activity of peroxidase and polyhenoloxidase enzymes in JA tubers than those of the uninfected JA tubers control in the two different storage methods. On the contrary, the application of caraway essential oil to infected/uninfected JA tubers increased peroxidase and polyphenoloxidase and phenol content compared with the untreated-uninfected JA tubers in both methods. These results are in a line with those obtained by [22] who reported an increasing in peroxidase and polyhenoloxidase enzymes in potato fresh tubers when treated with caraway essential oil. Although regulatory mechanisms of plant enzyme complexes and the most enzymatic reactions are reduced at low temperature degree, JA tubers metabolism could continue at a slow rate even at minimum temperature (2°C) during cold storage. The enzymatic activation due to the exogenous application of caraway essential oil treatment could be directly related to its content of bioconstituents like carvone.
Mean activities of peroxidase, polyphenoloxidase enzymes and phenol content (% ± 2SD) of JA tubers treated with caraway essential oil and infected with S. rolfsii using two storage methods.
1 = storage JA tubers in polyethylene bags at 4°C and 2 = storage JA tubers in peat moss layer at 25°C.
C = untreated control, P = infected tubers with pathogen, O = treated JA tubers with caraway essential oil and O + P = infected tubers with pathogen and treated with caraway essential oil.
NA = not applicable due to full decay.
2.2 Inhibition of sprout growth and increase storability of processing potato by antisprouting agent
2.2.1 Methodology
2.2.1.1 Tuber material
Fresh local potato cv. Fridor and uniformly size of 60–80 mm in diameter (weighing 180–250 g) were selected without any sprouting in eyes and no anti-sprouting treatment was used. Each treatment was treated with natural and safe antisprouting agent and stored at ambient temperature (average: 35/15°C day/night and 70% RH) in Laboratory for 4 months.
2.2.1.2 Treatments
The experiment included seven treatments, which were as follows: Cymbopogon martini (rich in geraniol and geranyl acetate), C. flexuosus (rich in citral), C. winterianus (rich in rich in citronellal and citronellol), Ocimum sanctum (rich in rich in ketone and camphor), Carum carvi (rich in rich in carvone), Artemisia annua (rich in ketone camphor) and Lavendula officinalis (rich in linalool). The isolated terpenoids were purified by HPLC. Essential oils were purified by column chromatography and substantially pure compounds were used. Tubers dipped in emulsions of 8 mm concentration of each compound in distilled water and Tween 20 (6%) for 30 min after 1 month of harvest or at such time that the tubers begin to sprout.
2.2.2 Results and discussion
2.2.2.1 Sprouting, weight loss and dry matter content
All control tubers had significant values of sprouting and weight loss percentages at the end of storage period (Table 5). Geraniol and citral completely inhibited sprouting by 100%, decreased weight loss and increase tuber dry matter content in both seasons. Application of geranyl acetate inhibited sprouting by 95%. On the other hand, linalool and l-carvone had no significant effect on tuber sprouting. It has been reported that l-carvone and d-carvone displayed little or no inhibition of sprouting in potatoes [17]. Geraniol and citral have a high content in monoterpenes such as benzaldehyde, eugenol and thymol [23]. CIPC inhibited sprouting over 98.5%.
Treatments
Sprouting (%)
Weight loss (%)
Dry matter (%)
2012
2013
2012
2013
2012
2013
1. Control
100.0a
96.00a
25.12a
26.18a
21.65f
22.80e
2. CIPC
2.49e
1.20c
4.33e
2.80ef
23.60a–d
23.66d
3. Geranyl acetate
4.68d
4.33c
3.41f
4.65d
22.50ef
24.55ab
4. Geraniol
0.00f
0.00c
2.19h
1.45g
24.56a
25.30a
5. Camphor
6.92c
5.98c
2.88g
2.95ef
23.33b–e
24.38bc
6. Citral
0.00f
0.00c
1.51i
1.26g
24.00ab
24.95ab
7. Linalool
100.00a
72.00b
9.50b
8.00b
22.66de
23.60d
8. l-Carvone
70.58b
62.00b
9.50b
6.25c
22.80c–e
23.70cd
9. d-Carvone
72.00b
76.98b
8.03c
3.45e
22.90c–e
24.89 ab
10. d-Citronellol
2.89e
2.00c
6.75d
5.73c
23.60a−d
24.68ab
11. l-Citronellol
0.00f
0.00c
2.25gh
2.10fg
23.80a–c
24.55ab
Table 5.
Sprouting behavior characters and dry matter of potato tubers as affected by anti-sprouting agent during 2012 and 2013 seasons (after 4 months of storage period).
Means followed by the same letter(s) within each column do not significantly differ using Duncan?s multiple range test at the level of 5%; where, (a) refer to the highest mean values, and (h) refer to the lowest mean values according to Duncan Multiple Range Test.
Under this study condition, the beneficial effect of the applied anti-sprouting agent (geraniol and citral) on controlling tubers sprouting and increasing dry matter content could be associated with their similar advantages effect in preservation of their tubers starch, carbohydrates, sugars and amino acid content (Table 6). Suppression of sprouting and weight loss logically associated with maintenance of dry matter. Furthermore, monoterpenes acts as antioxidant and had a protective role against oxidative stress under normal conditions of storage.
Treatments
Reducing sugars (%)
Total free amino acids (%)
Peroxidase activity POD (%)
2012
2013
2012
2013
2012
2013
1. Control
4.29a
4.52a
0.352a
0.348a
56.77g
55.51g
2. CIPC
2.05c
3.18d
0.307ab
0.301ab
95.81b
94.63b
3. Geranyl acetate
1.39cd
3.93b
0.084bc
0.047c
79.75e
79.33e
4. Geraniol
1.24d
1.51f
0.030c
0.028c
97.33a
96.29a
5. Camphor
3.41b
3.48c
0.152a–c
0.153a–c
80.68e
80.26e
6. Citral
1.25d
1.52f
0.045c
0.045c
97.68a
96.46a
7. Linalool
4.07ab
4.13b
0.106bc
0.108bc
80.67e
79.06c
8. l-Carvone
3.81ab
1.83e
0.084bc
0.151a–c
81.67e
80.50e
9. d-Carvone
1.45 cd
1.68ef
0.146a–c
0.157a–c
77.55f
76.77f
10. d-Citronellol
1.76cd
1.54f
0.186a–c
0.187a–c
84.50d
83.62d
11. l-Citronellol
1.29d
1.58f
0.147a–c
0.059c
87.67c
86.65c
Table 6.
Reducing sugars, amino acids and peroxidase enzyme of potato tubers as affected by anti-sprouting agent during 2012 and 2013 seasons (after 4 months of storage period).
Means followed by the same letter(s) within each column do not significantly differ using Duncan?s multiple range test at the level of 5%; where, (a) refer to the highest mean values, and (g) refer to the lowest mean values according to Duncan Multiple Range Test.
2.2.2.2 Reducing sugars, amino acids and peroxidase POD activity
All storage treatments gave significant lower values on reducing sugars and amino acids content during two seasons of study as compared to the control (Table 6). In the ambient temperature, the lowest significant values of reducing sugars and amino acids content were found in tubers exposed to emulsion of geraniol and citral, without significant difference between the two treatments.
The monoterpenes rich in compounds had a potential role in preservation and maintenance of the stored tubers reserves, keeping the enzymatic activities in a minimal level and in more stable case thereby prolonged their dormancy period. Also, application of these treatments were highly effective in tuber protection against the degradable effects of oxidative stressful during high temperature storage conditions and accordance to the findings of [20] who indicated that monoterpenes and antioxidants tended to slow down the activity of carbohydrates, breakdown of protein and enzymatic activity as well as reduce respiration rate and metabolism enzyme. The role of POD in sprouting of potatoes was widely reported, particularly its degrading activity of IAA, and cytokinin which is considered an effective promote oxidative stress is of great importance and depending on the activation degree of peroxidase as affected by storage treatments.
2.2.2.3 Processing quality of potato fries and chips
All storage treatments and CIPC treatment at ambient temperature had significant differences on quality characters of potato chips and French fries, i.e., color, crispiness and taste in comparison with the control treatment (Table 7).
Treatments
Chips
French fries
Color
Taste
Crispness
Color
Taste
Crispness
2012
2013
2012
2013
2012
2013
2012
2013
2012
2013
2012
2013
1. Control
3.00e
3.33c
3.00d
3.33bc
4.33a–c
4.33a–c
3.33 de
3.00 d
3.33cd
4.00b–d
4.67ab
4.67 ab
2. CIPC
3.33de
3.33c
4.33a–c
4.33ab
4.33a–c
4.67ab
3.67 cde
3.33 cd
4.00a–c
4.33a–c
4.67ab
4.67 ab
3. Geranyl acetate
4.67ab
4.67ab
5.00a
4.67a
5.00a
5.00a
4.67ab
4.67ab
5.00a
4.67ab
5.00a
4.33 abc
4. Geraniol
5.00a
5.00a
5.00a
4.67a
5.00a
5.00a
4.67ab
4.67ab
5.00a
4.67ab
5.00a
4.33 a–c
5. Camphor
4.67ab
4.67ab
5.00a
4.67a
5.00a
5.00a
4.67ab
4.67ab
4.67 ab
4.67ab
5.00a
5.00a
6. Citral
5.00a
5.00a
5.00a
4.67a
5.00a
5.00a
5.00a
5.00a
4.67ab
5.00a
5.00a
5.00a
7. Linalool
4.67ab
4.67 ab
4.64ab
4.67a
5.00a
5.00a
4.67ab
4.67ab
4.67ab
4.67ab
5.00a
5.00a
8. l-Carvone
4.67ab
4.67ab
4.67ab
4.67a
5.00a
5.00a
4.67ab
4.67ab
5.00a
5.00a
5.00a
5.00a
9. d-Carvone
5.00a
5.00a
5.00a
4.67a
5.00a
5.00a
5.00a
5.00a
4.67ab
5.00a
5.00a
5.00a
10. d-Citronellol
4.00b–d
4.67ab
4.67ab
4.67a
4.67ab
4.67a
4.00b–d
4.00a–c
4.33a–c
4.33a–c
4.67ab
5.00a
11. l-Citronellol
4.33a–c
4.67ab
4.67ab
4.67a
4.67ab
4.67a
4.00b–d
4.33a–c
3.67b–d
4.33a–c
4.67ab
4.67ab
Table 7.
Quality processing of potato tubers as affected by anti-sprouting agent during 2012 and 2013 seasons (after 4 months of storage period).
Means followed by the same letter(s) within each column do not significantly differ using Duncan?s multiple range test at the level of 5%; where, (a) refer to the highest mean values, and (d) refer to the lowest mean values according to Duncan Multiple Range Test.
The same treatments prevented and blocked the accumulation of total sugars, and kept the reducing sugars and amino acids in optimize levels in the stored tubers at ambient temperature. This is true in the end of storage (4 months). Thus, we noticed the worst processing quality (dark potato chips and crispness with bad taste) of storage treatments due to the appearance of Millard reaction during frying process and the accumulation of reducing sugars and amino acids [23]. The same processing quality parameters were correlated with dry matter content (Table 8) and with amino acids content (Table 9) in both seasons. These results are in harmony with those previously obtained by [24]. Meanwhile, we also noticed that the best processing quality of basic constituents of essential oils produced chips, the optimization of reducing sugars and amino acids of their tubers thereby, the prevention of Millard reaction occurrence during frying processes and thus it turn reflects on best color, crispiness and taste.
Treatment
Seed germination index [%]
Seed germination [%]
Seedling length [cm]
Seedling vigor index [cm]
Yield [g]container/242 cm2
2012
2013
2012
2013
2012
2013
2012
2013
2012
2013
1
Water (control)
13.36e
12.96d
86.67c
86.0a
4.67c
4.00b
4.03c
3.44c
36.40e
34.20d
2
Hot water
14.61de
13.02d
93.78b
90.44bc
5.00c
4.80b
4.71c
4.33c
40.88de
37.21d
3
Fennel oil
22.01a
23.01a
97.33ab
97.33a
7.33ab
7.67a
7.13ab
7.47ab
56.90b
49.17c
4
Caraway oil
21.94a
22.88a
97.33ab
99.00a
8.00ab
8.33a
7.79a
8.25a
54.97bc
67.75a
5
Basil oil
20.22ab
21.82ab
94.67b
92.33b
7.00b
7.67a
6.63b
7.07b
64.87a
68.17a
6
Thyme oil
18.81bc
20.14b
100.00a
100.0a
8.20a
8.30a
8.20a
8.30a
66.54a
67.75a
7
Sage oil
16.91cd
17.76c
100.00a
100.0a
7.83ab
7.83a
7.83a
7.83ab
47.83cd
49.17c
Table 8.
Vegetative characters of treated broccoli seeds with different essential oils before cold storage.
Means followed by the same letter(s) within each column do not significantly differ using Duncan?s multiple range test at the level of 5%; where, (a) refer to the highest mean values, and (e) refer to the lowest mean values according to Duncan Multiple Range Test.
Treatment
Total phenolic acid (mg/100 g F.W.)
Total flavonoids (mg/100 g F.W.)
Anthocyanin (mg/100 g F.W.)
Ascorbic acid (mg/100 g F.W.)
DPPH (Mmol TE/g F.W.)
2012
2013
2012
2013
2012
2013
2012
2013
2012
2013
1
Water (control)
83.33d
84.11e
91.99d
95.18e
7.13d
7.70d
70.58e
81.23d
23.66a
24.66a
2
Hot water
88.71c
88.56c
100.95c
101.03d
8.62c
8.77c
86.81c
86.81c
23.54b
23.66a
3
Fennel oil
88.46c
88.90c
107.66b
107.72c
8.86c
8.87bc
87.66c
88.00c
21.98d
21.98c
4
Caraway oil
87.90c
88.13cd
104.66b
104.73c
9.84bc
9.84bc
77.33d
85.80c
21.96de
21.96c
5
Basil oil
122.06b
122.29b
113.00a
113.00b
11.71a
12.05a
94.67b
94.67b
21.94de
21.94c
6
Thyme oil
131.66a
131.60a
115.66a
116.24a
12.09a
12.14a
102.33a
103.33a
21.86e
20.03d
7
Sage oil
87.9c
84.74de
104.33bc
104.59c
10.38b
10.38b
82.33cd
86.69c
22.79c
22.79bc
Table 9.
Phytochemical screening by GLC for 3-days-old broccoli sprouts produced from treated seeds with essential oils before cold storage.
Means followed by the same letter(s) within each column do not significantly differ using Duncan?s multiple range test at the level of 5%; where, (a) refer to the highest mean values, and (e) refer to the lowest mean values according to Duncan Multiple Range Test.
2.3 Increasing antioxidant content of broccoli sprouts using essential oils during cold storage
2.3.1 Methodology
2.3.1.1 Plant material and germination condition
(Brassica oleracea L. var. italica and the variety name is F1 Hybrid Sakura) from Tokita Seeds CO., LTD (Saitama, Japan). The seeds (1000 seeds, nearly 5 g) for each treatment were soaked in a sodium hypochlorite solution at 0.5% v/v for 15 min then were dipped in 50 ml of deionized water for 5½ h with shaking every 30 min and washed with deionized and sterilized water. On 15th of September, broadcast the seeds were done over absorbent medical cotton in sprouting plastic containers (220 × 110 mm). The emulsions of various natural essential oil at the concentration of 0.05% were emulsified in tween 80 (1.5 ml/l) in the cotton media and the containers were getting closed immediately. The containers were maintained at 25 ± 2°C with and 16 h light/8 h darkness, 80–90% relative humidity and 7.4 lmol/m2/s light intensity to give the best germination conditions. All sprouts in containers were cut above their root mats after 3 days from sowing. The sprouts were weighed for 20 g for each placed container and stored at 4°C in the dark to simulate a domestic refrigerator for 15 days. The sprouts of best treatment with control were stored only.
2.3.1.2 Application and extraction of essential oils
The essential oils of fennel seeds (Foeniculum vulgare), caraway seeds (Carum carvi), thyme herbs (Thymus vulgaris), basil herbs (Ocimum basilicum) and sage leaves (Salvia officinalis) (200 g from each one) were used for oil extraction by hydro-distillation for 2–3 h. After extraction, essential oils were analyzed by Gas Liquid Chromatography (GLC) to separate and identify their basic constituents.
2.3.2 Results and discussion
2.3.2.1 Vegetative characters of broccoli sprout
All essential oil treatments rich in antioxidant stimulate the germination of broccoli seeds. All essential oils treatments significantly increased germination, germination index, seedling length, seedling vigor index and container yield compared with the control (tap water) during the two seasons (Table 8). The essential oils of fennel, caraway and thyme increased the seed germination index by 171.43, 170.29 and 148.02%, respectively, compared to the control 100%. The increases of seed germination % over the control reached to 12.73, 13.74 and 15.82% for the effective treatments, respectively. The essential oils of thyme, caraway and fennel had significant increases in seedling vigor and yield container over the control to 50.25, 73.82 and 90.22%, respectively.
The allelochemical effects of essential oils for induce stimulatory or inhibitory of seed germination and other physiological process varied depending on the dose, tested species, concentration and basic components. Under our study, the lower doses of essential oils had a stimulatory effect [25]. The obtained results reveal that the applications of essential oils at a low level improve seed germination of broccoli. However, application of thyme oil reaches 100% of sprouts after seed germination (Table 8). Impact of essential oils on seed germination of other plant species was reported as 24 out of 47 tested terpenoids enhanced the seed germination of Lactuca sativa [26]. Also, the positive impact of thyme essential oil on broccoli seeds could be because of its active ingredients.
2.3.2.2 Phytochemical characters
All treatments significantly surpassed over the control in Broccoli sprout bio-constituents, i.e., total phenolic acid, total flavonoids, anthocyanin and ascorbic acid, while the control treatment gave the highest DPPH radical scavenging capacity (Table 9). Application of thyme oil treatment produced significant increases of total phenol, total flavonoids, anthocyanin and ascorbic acid content. Moreover, thyme and basil essential oils decreased significantly the DPPH free radical scavenging capacity. Accordingly, it has been chosen to study the storage behavior characters, in addition to control treatment. The majority of the antioxidant activity attributes to phenolic compounds, flavonoids and ascorbic acidin essential oils [27]. Moreover, the effect of antioxidant on DPPH free radicle was due to the presence of hydroxyl groups in their chemical structure. In this respect, [28] found that the oregano essential oil inhibited hydro-peroxide formation and that the CHO fraction showed the highest antioxidants activity.
The thyme oil showed significant lowest radical scavenging capacity compared to the control and other treatments (Table 9). All other antioxidants/essential oils showed high and almost the same antioxidant capacity effect. It was known that the free radical scavenging DPPH intensity of some compounds can be influenced by their different kinetic behavior [29]. For slow reacting compounds the influence was attributed to the complex reacting mechanism. In our study, probably, the constituents from thyme essential oil involved one or more secondary reactions, which result the slower reduction of DPPH solutions [29].
2.3.2.3 Antioxidant activity during cold storage
2.3.2.3.1 Total phenolic compounds and DPPH radical scavenging capacity
Figure 2 illustrate that there was a gradual increase in the total phenolic acid content, and reaching a maximum value at day 5 and 10 (132.67 and 135.04 mg GAE/100 g F.W.) compared to the initial time. This concentration decreased in to 129.03 mg at day 15 due to thyme oil application (Figure 2). Keeping in view that the control treatment decreased to 73.84 GAE/100 g FW at day 5. On the 15th day, the old-sprout from storage, the control was reduced by 28.57% compared to thyme oil (1.98%). The control treatment of antioxidant capacity increased significantly until day 10 (29.43 mg/100 g F.W.), and finally decrease (28.46% mg 100/g F.W.) at day 15 increased from initial period (20.28%). While, application of thyme oil the change was not clear at the end of storage (1.98%) (Figure 3). During cold storage (Figure 3), the control was reduced DPPH by 28.57% compared to thyme oil at 15 day old-sprout (1.98%). Nath et al. [30] observed a constant decrease in the antioxidant capacity for 144 h of storage of broccoli inflorescences. This behavior in DPPH may be due to the steady changes in plant metabolism during storage period as a result of oxidative stress, which may include structural and chemical changes in synthesis or antioxidant content [31].
Figure 2.
Total phenolic content as affected by thyme oil compared to control treatment at different storage period.
Figure 3.
DPPH radical scavenging capacity as affected by thyme oil compared to control treatment at different storage period.
2.3.2.3.2 Total flavonoids
Total flavonoids (Figure 4) were found in a higher concentration in 3-day-old sprouts of thyme treatment, with values of 115.95 mg/100 g F.W., after 5 and 10 days of storage slight decrease to 0.021 and 0.086%, respectively, when compared with the initial value, and finally reduced by 1.39%. The high loss of flavonoids reached to 10.59 and 47.89%, after 5 and 10 days, respectively, and at 15 days the loss increased to 58.33% for control treatment (average two seasons).
Figure 4.
Total flavonoids content as affected by thyme oil compared to control treatment at different storage period.
2.3.2.3.3 Glucosinolates content
Storage time had significant differences in glucosinolates content of the samples analyzed. Figure 5 illustrate that the thyme oil increased significantly glucosinolates content in 3-day-old sprouts, compared to control treatments. Moreover, thyme oil had a high value of total glucosinolates (27.02 μg/g F.W.) and slightly decreased up to 26.43 μg/g F.W. on day 15. At the end of storage, the decreasing changes percent was about 2.18%. In the control treatment, the highest decrease in total glucosinolates content was observed, where reached about 49.12% at the end of storage.
Figure 5.
Total glucosinolates content as affected by thyme oil compared to control treatment at 0 time and 15 DAS.
2.3.2.3.4 Glucosinolates content of mature head versus sprout broccoli
In sprout, the total glucosinolates level (27.02 μg/g F.W.) is higher than in florets or heads (7.37) (Figure 6). Glucoraphanin is the powerful of antioxidant and the most abundant aliphatic glucosinolates present in sprout. The glucoraphanin reached the highest 16.24 followed by glucoerucin 5.9 and glucoiberm 1.2 μg/g F.W. On the other hand, the florets/heads contain the highest level of aromatic/indolylglucosinolates, neoglucobrassicin (2.11) followed by glucobrassicin (1.67). Our results are in agreement with those obtained by [32].
Figure 6.
Total and individual aliphatic, aromatic/indole glucosinolates levels in broccoli in 3-days-old sprout and mature at harvest.
\n',keywords:"essential oils, constituents, anti-sprouting agent, antifungal, sprout growth, postharvest, vegetables, structure, extraction, application",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/67937.pdf",chapterXML:"https://mts.intechopen.com/source/xml/67937.xml",downloadPdfUrl:"/chapter/pdf-download/67937",previewPdfUrl:"/chapter/pdf-preview/67937",totalDownloads:51,totalViews:0,totalCrossrefCites:0,dateSubmitted:"December 24th 2018",dateReviewed:"June 3rd 2019",datePrePublished:"August 21st 2019",datePublished:null,readingETA:"0",abstract:"Essential oils, as natural sprout inhibitor and safe fungicides, are a promising tool and good alternative compounds otherwise synthetic due to their high efficacy, biodegradability, eco-safety and volatile nature. They are consisting of a number of various components, i.e., terpenes, phenols, alcohols, esters, aldehydes and ketones in different composition or combinations. These effective compounds supply excess to prevent sprouting in potatoes and Jerusalem artichoke (JA) and less chance to development of resistance in fungi in JA, strawberry and broccoli with low concentrations. On contrary, high concentration of these oils induce the germination of seeds like broccoli and carob. This chapter explains the practical application of using essential oils as natural antisprouting, inducing quality, preserving fungal diseases, eco-friendly compounds, alternating synthetic chemicals, giving high benefits and easy to apply. The foliar application with essential oils increases the productivity, quality and marketable yield and storability and reduces weight losses and decay. Moreover, the essential oils increase broccoli seed germination, antioxidant content and other phytochemical parameters. The chapter provides a novel anti-sprouting agent for inhibiting growth of processing potato tubers and identification of terpenoids that use to inhibit tuber sprouting as well as application of Chloropropham (CIPC) isopropyl-N-(3-chlorophenyl) carbamate as a conventional chemical inhibitor.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/67937",risUrl:"/chapter/ris/67937",signatures:"Aml Abo El-Fetouh El-Awady",book:{id:"7855",title:"Essential Oils - Oils of Nature",subtitle:null,fullTitle:"Essential Oils - Oils of Nature",slug:null,publishedDate:null,bookSignature:"Prof. Hany El-Shemy",coverURL:"https://cdn.intechopen.com/books/images_new/7855.jpg",licenceType:"CC BY 3.0",editedByType:null,editors:[{id:"54719",title:"Prof.",name:"Hany",middleName:null,surname:"El-Shemy",slug:"hany-el-shemy",fullName:"Hany El-Shemy"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:null,sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Application of essential oils",level:"1"},{id:"sec_2_2",title:"2.1 Alternative preservation method against sclerotium tuber rot of Jerusalem artichoke using natural essential oils",level:"2"},{id:"sec_2_3",title:"2.1.1 Methodology",level:"3"},{id:"sec_3_3",title:"Table 1.",level:"3"},{id:"sec_3_4",title:"2.1.2.1 Antifungal activity of the essential oils",level:"4"},{id:"sec_4_4",title:"Table 1.",level:"4"},{id:"sec_4_5",title:"Table 1.",level:"5"},{id:"sec_5_5",title:"Table 2.",level:"5"},{id:"sec_6_5",title:"Table 3.",level:"5"},{id:"sec_7_5",title:"Table 4.",level:"5"},{id:"sec_11_2",title:"2.2 Inhibition of sprout growth and increase storability of processing potato by antisprouting agent",level:"2"},{id:"sec_11_3",title:"2.2.1 Methodology",level:"3"},{id:"sec_11_4",title:"2.2.1.1 Tuber material",level:"4"},{id:"sec_12_4",title:"2.2.1.2 Treatments",level:"4"},{id:"sec_14_3",title:"Table 5.",level:"3"},{id:"sec_14_4",title:"Table 5.",level:"4"},{id:"sec_15_4",title:"2.2.2.2 Reducing sugars, amino acids and peroxidase POD activity",level:"4"},{id:"sec_16_4",title:"Table 7.",level:"4"},{id:"sec_19_2",title:"2.3 Increasing antioxidant content of broccoli sprouts using essential oils during cold storage",level:"2"},{id:"sec_19_3",title:"2.3.1 Methodology",level:"3"},{id:"sec_19_4",title:"2.3.1.1 Plant material and germination condition",level:"4"},{id:"sec_20_4",title:"2.3.1.2 Application and extraction of essential oils",level:"4"},{id:"sec_22_3",title:"2.3.2 Results and discussion",level:"3"},{id:"sec_22_4",title:"2.3.2.1 Vegetative characters of broccoli sprout",level:"4"},{id:"sec_23_4",title:"2.3.2.2 Phytochemical characters",level:"4"},{id:"sec_24_4",title:"2.3.2.3 Antioxidant activity during cold storage",level:"4"},{id:"sec_24_5",title:"2.3.2.3.1 Total phenolic compounds and DPPH radical scavenging capacity",level:"5"},{id:"sec_25_5",title:"2.3.2.3.2 Total flavonoids",level:"5"},{id:"sec_26_5",title:"2.3.2.3.3 Glucosinolates content",level:"5"},{id:"sec_27_5",title:"2.3.2.3.4 Glucosinolates content of mature head versus sprout broccoli",level:"5"}],chapterReferences:[{id:"B1",body:'Tesio F, Weston LA, Ferrero A. Allelochemicals identified from Jerusalem artichoke (Helianthus tuberosus L.) residues and their potential inhibitory activity in the field and laboratory. Scientia Horticulturae Amsterdam. 2011;129:361-368. DOI: 10.1016/j.scienta.2011.04.003'},{id:"B2",body:'Yang L, He QS, Corscadden K, Udenigwe CC. The prospects of Jerusalem artichoke in functional food ingredients and bioenergy production. Biotechnology Reports. 2015;5:77-88. DOI: 10.1016/j.btre.2014.12.004'},{id:"B3",body:'Zhang Y, Li S, Jiang D, Kong L, Zhang P, Xu J. Antifungal activities of metabolites produced by a termite-associated Streptomyces canus BYB02. Journal of Agricultural and Food Chemistry. 2013;61:1521-1524. DOI: 10.1021/jf305210u'},{id:"B4",body:'Suhag M, Nehra BK, Singh N, Khurana SC. Storage behavior of potato under ambient condition affected by curing and crop duration. Haryana Journal Of Horticultural Sciences. 2006;35:357-360'},{id:"B5",body:'Delaplace P, Brostaux Y, Fauconnier ML, du Jardin P. Potato (Solanum tuberosum L.) tuber physiological age index is a valid reference frame in postharvest ageing studies. Postharvest Biology and Technology. 2008;50:103-106. DOI: 10.1093/jxb/erp008'},{id:"B6",body:'De Carvalho C, Da Fonseca MMR. Carvone: Why and how should one bother to produce this terpene. Food Chemistry. 2006;95:413-422. DOI: 10.1016/j.foodchem.2005.01.003'},{id:"B7",body:'Pringle B, Bishop C, Clayton R. Potatoes Postharvest. UK: CAB International; 2009. p. 427'},{id:"B8",body:'Villarreal-Garcia D, Nair V, Gisneros-Zevallos L, Jacobo-Veazquez DA. Plants as biofacttories: Postharvest stress-induced accumulation of phenolic compounds and glucosinolates in broccoli subjected to wounding stress and exogenous phytohormones. In Fronties in Plant Science. 2016;7(45):1-11. DOI: 10.3389/fpls.2016.00045'},{id:"B9",body:'Baenas N, García-Viguera C, Moreno DA. Biotic elicitors effectively increase the glucosinolates content in brassicaceae sprouts. Journal of Agricultural and Food Chemistry. 2014;62:1881-1889. DOI: 10.1021/jf404876z'},{id:"B10",body:'Nguefack J, Somda I, Mortensen CN, Amvam Zollo PH. Evaluation of five essential oils from aromatic plants of Cameroon for controlling seed-borne bacteria of rice (Oryza sativa L.). Seed Science and Technology. 2005;33:397-407. DOI: 10.15258/sst.2005.33.2.12'},{id:"B11",body:'Justesen U, Knuthsen P. Composition of flavonoids in fresh herbs and calculation of flavonoid intake by use of herbs in traditional Danish dishes. Food Chemistry. 2001;73:245-250'},{id:"B12",body:'Darougheh F, Barzegar M, Sahari M. Antioxidant and anti-fungal effect of caraway (Carum carvi L.). Essential oil in real food system. Current Nutrition & Food Science. 2014;10(1):70-76'},{id:"B13",body:'Ma B, Ban X, Huang B, He J, Tian J, Zeng H, et al. Interference and mechanism of dill seed essential oil and contribution of carvone and limonene in preventing Sclerotinia rot of rapeseed. PLoS ONE. 2015;10(7):e0131733. DOI: 10.1371/journal.pone.0131733'},{id:"B14",body:'Taskila S, Särkelä R, Tanskanen J. Valuable applications for peat moss. Biomass Conversion and Biorefinery. 2016;6:115-126'},{id:"B15",body:'Zaitseva N. A polysaccharide extracted from Sphagnum moss as antifungal agent in archaeological conservation [master’s thesis]. Ontario, Canada: Queen’s University, Kingston; 2009. p. 282'},{id:"B16",body:'Borsheim KY, Christensen BE, Painter T. Preservation of fish by embedment in sphagnum moss, peat, or holocellulose: Experimental proof of the oxopolysaccharidic nature of the preservative substance and its antimicrobial and tanning action. Innovative Food Science and Emerging Technologies. 2012;2(1):63-74'},{id:"B17",body:'Oosterhaven K, Hartmans KJ, Scheffer JJC. Inhibition of potato sprouts growth by carvone enantiomers and their bioconversion in sprout. Potato Research. 1995;38:219-230'},{id:"B18",body:'Cabezas MJ, Rabert C, Bravo S, Shene C. Inulin and sugar contents in Helianthus tuberosus and Cichorium intybus tubers: Effect of post-harvest storage temperature. Journal of Food Science. 2002;67:2860-2865'},{id:"B19",body:'Brkljaca J, Bodroza-Solarov M, Krulj J, Terzic S, Mikic A, Marjanovic-Jeromela A. Quantification of inulin content in selected accessions of Jerusalem artichoke (Helianthus tuberosus L.). Helia. 2014;37(60):105-112'},{id:"B20",body:'Norkulova KT, Safarov JE. Research of sorption characteristics of tubers Jerusalem artichoke (Helianthus tuberosus). Journal of Food Processing and Technology. 2015;6(6):453-454'},{id:"B21",body:'Davies HV. Carbohydrate metabolism during sprouting. American Potato Journal. 1990;67:815-827'},{id:"B22",body:'Afify AMR, El-Beltagi HS, Aly AA, El-Ansary AE. Antioxidant enzyme activities and lipid peroxidation as biomarker for potato tuber stored by two essential oils from Caraway and Clove and its main component carvone and eugenol. Asian Pacific Journal of Tropical Biomedicine. 2012;2:S772-S780'},{id:"B23",body:'Hartmans KJ, Diepenhorst P, Bakker W, Gorris LGM. The use of carvone in agriculture: Sprout suppression of potatoes and antifungal activity against potato tuber and other plant diseases. Industrial Crops and Products. 1995;4:3-13'},{id:"B24",body:'El-Awady AA. Studies on storing potato tubers out refrigerator using natural essential oils [Ph.D. thesis]. Fac. Agriculture: Mansoura University; 2006. p. 166'},{id:"B25",body:'Leth V. Use of essential oils as seed treatment. IPGRI Newsletter. 2002;9:15-16'},{id:"B26",body:'Vokou D, Douvli P, Blionis GJ, Halley JM. Effects of monoterpenoids, acting alone or in pairs, on seed germination and subsequent seedling growth. Journal of Chemical Ecololgy. 2003;29:2281-2301. DOI: 10.1023/A:1026274430898'},{id:"B27",body:'Heim KE, Tagliaferro AR, Bobilya DJ. Flavonoid antioxidants: Chemistry, metabolism and structure-activity relationships. The Journal of Nutritional Biochemistry. 2002;13:572-584. DOI: 10.1016/S0955-2863(02)00208-5'},{id:"B28",body:'Milos M, Mastelic J, Jerkovic I. Chemical composition and antioxidant effect of glycosidically bound volatile compound from oregano (Origanum vulgare L. ssp. hirtum). Food Chemistry. 2000;71:79-83'},{id:"B29",body:'Konczak I, Zhang W. Anthocyanins—More than nature’s colours. Journal of Biomedicine Biotechnology. 2004;5:239-240. DOI: 10.1155S1110724304407013'},{id:"B30",body:'Nath A, Bagchi B, Misra LK, Deka BC. Changes in post-harvest phytochemical qualities of broccoli florets during ambient and refrigerated storage. Food Chemistry. 2011;127:1510-1514. DOI: 10.1016/j.foodchem.2011.02.007'},{id:"B31",body:'Xiao Z, Lester GE, Luo Y, Xie Z, Yu L, Wang Q. Effect of light exposure on sensorial quality, concentrations of bioactive compounds and antioxidant capacity of radish microgreens during low temperature storage. Food Chemistry. 2014;151:472-479. DOI: 10.1016/j.foodchem.2013.11.086'},{id:"B32",body:'Fahey JW, Zhang Y, Talalay P. Broccoli sprouts: An exceptionally rich source of inducers of enzymes that protect against chemical carcinogens. Proceedings National Academy of Sciences of the United States of America, USA. 1997;94:10367-10372'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Aml Abo El-Fetouh El-Awady",address:"aml.elawady@yahoo.com",affiliation:'
Horticulture Research Institute, Agriculture Research Center, Giza, Egypt
'}],corrections:null},book:{id:"7855",title:"Essential Oils - Oils of Nature",subtitle:null,fullTitle:"Essential Oils - Oils of Nature",slug:null,publishedDate:null,bookSignature:"Prof. Hany El-Shemy",coverURL:"https://cdn.intechopen.com/books/images_new/7855.jpg",licenceType:"CC BY 3.0",editedByType:null,editors:[{id:"54719",title:"Prof.",name:"Hany",middleName:null,surname:"El-Shemy",slug:"hany-el-shemy",fullName:"Hany El-Shemy"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}}},profile:{item:{id:"247878",title:"Dr.",name:"Kapil",middleName:null,surname:"Sharma",email:"kapil05vats@gmail.com",fullName:"Kapil Sharma",slug:"kapil-sharma",position:null,biography:null,institutionString:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",totalCites:0,totalChapterViews:"0",outsideEditionCount:0,totalAuthoredChapters:"1",totalEditedBooks:"0",personalWebsiteURL:null,twitterURL:null,linkedinURL:null,institution:null},booksEdited:[],chaptersAuthored:[{title:"Plant Metabolomics: An Emerging Technology for Crop Improvement",slug:"plant-metabolomics-an-emerging-technology-for-crop-improvement",abstract:"The astounding ability of plants to make smart decisions in response to environment is evident. As they have evolved a long list of complex and unique processes that involve photosynthesis, totipotency, long-distance signaling, and ability to restore structural and metabolic memory, recognition, and communication via emission of the selected class of volatiles. In recent years, use of metabolite profiling techniques in detection, unambiguous identification, quantification, and rapid analysis of the minute quantity of cellular micromolecules has increased considerably. Metabolomics is key to understand the chemical footprints during different phases of growth and development of plants. To feed the ever-increasing population with limited inputs and in a rapidly changing environment is the biggest challenges that the world agriculture faces today. To achieve the project genetic gains, the breeding strategies employing marker-assisted selection for high-yielding varieties and identifying germplasm resistant to abiotic and biotic stresses are already in vogue. Henceforth, new approaches are needed to discover and deploy agronomically important gene/s that can help crops better withstand weather extremes and growing pest prevalence worldwide. In this context, metabolic engineering technology looks viable option, with immense potential to deliver the future crops.",signatures:"Kapil Sharma, Supriya Sarma, Abhishek Bohra, Abhijit Mitra,\nNaveen K. Sharma and Anirudh Kumar",authors:[{id:"237447",title:"Dr.",name:"Anirudh",surname:"Kumar",fullName:"Anirudh Kumar",slug:"anirudh-kumar",email:"anirudh.kumar@igntu.ac.in"},{id:"247878",title:"Dr.",name:"Kapil",surname:"Sharma",fullName:"Kapil Sharma",slug:"kapil-sharma",email:"kapil05vats@gmail.com"},{id:"247879",title:"Dr.",name:"Supriya",surname:"Sharma",fullName:"Supriya Sharma",slug:"supriya-sharma",email:"dipu.megha@gmail.com"},{id:"247880",title:"Dr.",name:"Abhishek",surname:"Bohra",fullName:"Abhishek Bohra",slug:"abhishek-bohra",email:"abhi.omics@gmail.com"},{id:"247882",title:"Prof.",name:"Abhijit",surname:"Mitra",fullName:"Abhijit Mitra",slug:"abhijit-mitra",email:"abi_chem@iiit.ac.in"},{id:"247884",title:"Prof.",name:"Naveen K.",surname:"Sharma",fullName:"Naveen K. Sharma",slug:"naveen-k.-sharma",email:"nks.igntu@gmail.com"}],book:{title:"New Visions in Plant Science",slug:"new-visions-in-plant-science",productType:{id:"1",title:"Edited Volume"}}}],collaborators:[{id:"213506",title:"Dr.",name:"Lakesh",surname:"Sharma",slug:"lakesh-sharma",fullName:"Lakesh Sharma",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Maine",institutionURL:null,country:{name:"United States of America"}}},{id:"230515",title:"Assistant Prof.",name:"Zhong",surname:"Chen",slug:"zhong-chen",fullName:"Zhong Chen",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"233336",title:"Dr.",name:"Sukhwinder",surname:"K. Bali",slug:"sukhwinder-k.-bali",fullName:"Sukhwinder K. Bali",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"233337",title:"Dr.",name:"Ahmed",surname:"Zaeen",slug:"ahmed-zaeen",fullName:"Ahmed Zaeen",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"233338",title:"Dr.",name:"James",surname:"Dwyer",slug:"james-dwyer",fullName:"James Dwyer",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"237447",title:"Dr.",name:"Anirudh",surname:"Kumar",slug:"anirudh-kumar",fullName:"Anirudh Kumar",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"239423",title:"Dr.",name:"An",surname:"Yan",slug:"an-yan",fullName:"An Yan",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"247879",title:"Dr.",name:"Supriya",surname:"Sharma",slug:"supriya-sharma",fullName:"Supriya Sharma",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"247880",title:"Dr.",name:"Abhishek",surname:"Bohra",slug:"abhishek-bohra",fullName:"Abhishek Bohra",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"247882",title:"Prof.",name:"Abhijit",surname:"Mitra",slug:"abhijit-mitra",fullName:"Abhijit Mitra",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null}]},generic:{page:{slug:"OA-publishing-fees",title:"Open Access Publishing Fees",intro:"
The Open Access model is applied to all of our publications and is designed to eliminate subscriptions and pay-per-view fees. This approach ensures free, immediate access to full text versions of your research.
As a gold Open Access publisher, an Open Access Publishing Fee is payable on acceptance following peer review of the manuscript. In return, we provide high quality publishing services and exclusive benefits for all contributors. IntechOpen is the trusted publishing partner of over 116,000 international scientists and researchers.
\\n\\n
The Open Access Publishing Fee (OAPF) is payable only after your full chapter, monograph or Compacts monograph is accepted for publication.
\\n\\n
OAPF Publishing Options
\\n\\n
\\n\\t
1,400 GBP Chapter - Edited Volume
\\n\\t
10,000 GBP Monograph - Long Form
\\n\\t
4,000 GBP Compacts Monograph - Short Form
\\n
\\n\\n
*These prices do not include Value-Added Tax (VAT). Residents of European Union countries need to add VAT based on the specific rate in their country of residence. Institutions and companies registered as VAT taxable entities in their own EU member state will not pay VAT as long as provision of the VAT registration number is made during the application process. This is made possible by the EU reverse charge method.
\\n\\n
Services included are:
\\n\\n
\\n\\t
An online manuscript tracking system to facilitate your work
\\n\\t
Personal contact and support throughout the publishing process from your dedicated Author Service Manager
\\n\\t
Assurance that your manuscript meets the highest publishing standards
\\n\\t
English language copyediting and proofreading, including the correction of grammatical, spelling, and other common errors
\\n\\t
XML Typesetting and pagination - web (PDF, HTML) and print files preparation
\\n\\t
Discoverability - electronic citation and linking via DOI
\\n\\t
Permanent and unrestricted online access to your work
What isn't covered by the Open Access Publishing Fee?
\\n\\n
If your manuscript:
\\n\\n
\\n\\t
Exceeds 20 pages (for chapters in Edited Volumes), an additional fee of 40 GBP per page will be required
\\n\\t
If a manuscript requires Heavy Editing or Language Polishing, this will incur additional fees.
\\n
\\n\\n
Your Author Service Manager will inform you of any items not covered by the OAPF and provide exact information regarding those additional costs before proceeding.
\\n\\n
Open Access Funding
\\n\\n
To explore funding opportunities and learn more about how you can finance your IntechOpen publication, go to our Open Access Funding page. IntechOpen offers expert assistance to all of its Authors. We can support you in approaching funding bodies and institutions in relation to publishing fees by providing information about compliance with the Open Access policies of your funder or institution. We can also assist with communicating the benefits of Open Access in order to support and strengthen your funding request and provide personal guidance through your application process. You can contact us at oapf@intechopen.com for further details or assistance.
\\n\\n
For Authors who are still unable to obtain funding from their institutions or research funding bodies for individual projects, IntechOpen does offer the possibility of applying for a Waiver to offset some or all processing feed. Details regarding our Waiver Policy can be found here.
\\n\\n
Added Value of Publishing with IntechOpen
\\n\\n
Choosing to publish with IntechOpen ensures the following benefits:
\\n\\n
\\n\\t
Indexing and listing across major repositories
\\n\\t
Long-term archiving Visibility on the world's strongest OA platform
\\n\\t
Live Performance Metrics to track readership and the impact of your chapter
\\n\\t
Dissemination and Promotion
\\n
\\n\\n
Benefits of Publishing with IntechOpen
\\n\\n
\\n\\t
Proven world leader in Open Access book publishing with over 10 years experience
\\n\\t
+4,400 OA books published
\\n\\t
Most competitive prices in the market
\\n\\t
Fully compliant with OA funding requirements
\\n\\t
Optimized processes, enabling publication between 8 and 12 months
\\n\\t
Personal support during every step of the publication process
\\n\\t
+84,800 citations in Web of Science databases
\\n\\t
Currently strongest OA platform with over 130 million downloads
As a gold Open Access publisher, an Open Access Publishing Fee is payable on acceptance following peer review of the manuscript. In return, we provide high quality publishing services and exclusive benefits for all contributors. IntechOpen is the trusted publishing partner of over 116,000 international scientists and researchers.
\n\n
The Open Access Publishing Fee (OAPF) is payable only after your full chapter, monograph or Compacts monograph is accepted for publication.
\n\n
OAPF Publishing Options
\n\n
\n\t
1,400 GBP Chapter - Edited Volume
\n\t
10,000 GBP Monograph - Long Form
\n\t
4,000 GBP Compacts Monograph - Short Form
\n
\n\n
*These prices do not include Value-Added Tax (VAT). Residents of European Union countries need to add VAT based on the specific rate in their country of residence. Institutions and companies registered as VAT taxable entities in their own EU member state will not pay VAT as long as provision of the VAT registration number is made during the application process. This is made possible by the EU reverse charge method.
\n\n
Services included are:
\n\n
\n\t
An online manuscript tracking system to facilitate your work
\n\t
Personal contact and support throughout the publishing process from your dedicated Author Service Manager
\n\t
Assurance that your manuscript meets the highest publishing standards
\n\t
English language copyediting and proofreading, including the correction of grammatical, spelling, and other common errors
\n\t
XML Typesetting and pagination - web (PDF, HTML) and print files preparation
\n\t
Discoverability - electronic citation and linking via DOI
\n\t
Permanent and unrestricted online access to your work
What isn't covered by the Open Access Publishing Fee?
\n\n
If your manuscript:
\n\n
\n\t
Exceeds 20 pages (for chapters in Edited Volumes), an additional fee of 40 GBP per page will be required
\n\t
If a manuscript requires Heavy Editing or Language Polishing, this will incur additional fees.
\n
\n\n
Your Author Service Manager will inform you of any items not covered by the OAPF and provide exact information regarding those additional costs before proceeding.
\n\n
Open Access Funding
\n\n
To explore funding opportunities and learn more about how you can finance your IntechOpen publication, go to our Open Access Funding page. IntechOpen offers expert assistance to all of its Authors. We can support you in approaching funding bodies and institutions in relation to publishing fees by providing information about compliance with the Open Access policies of your funder or institution. We can also assist with communicating the benefits of Open Access in order to support and strengthen your funding request and provide personal guidance through your application process. You can contact us at oapf@intechopen.com for further details or assistance.
\n\n
For Authors who are still unable to obtain funding from their institutions or research funding bodies for individual projects, IntechOpen does offer the possibility of applying for a Waiver to offset some or all processing feed. Details regarding our Waiver Policy can be found here.
\n\n
Added Value of Publishing with IntechOpen
\n\n
Choosing to publish with IntechOpen ensures the following benefits:
\n\n
\n\t
Indexing and listing across major repositories
\n\t
Long-term archiving Visibility on the world's strongest OA platform
\n\t
Live Performance Metrics to track readership and the impact of your chapter
\n\t
Dissemination and Promotion
\n
\n\n
Benefits of Publishing with IntechOpen
\n\n
\n\t
Proven world leader in Open Access book publishing with over 10 years experience
\n\t
+4,400 OA books published
\n\t
Most competitive prices in the market
\n\t
Fully compliant with OA funding requirements
\n\t
Optimized processes, enabling publication between 8 and 12 months
\n\t
Personal support during every step of the publication process
\n\t
+84,800 citations in Web of Science databases
\n\t
Currently strongest OA platform with over 130 million downloads
\n
\n'}]},successStories:{items:[]},authorsAndEditors:{filterParams:{sort:"featured,name"},profiles:[{id:"6700",title:"Dr.",name:"Abbass A.",middleName:null,surname:"Hashim",slug:"abbass-a.-hashim",fullName:"Abbass A. Hashim",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/6700/images/1864_n.jpg",biography:"Currently I am carrying out research in several areas of interest, mainly covering work on chemical and bio-sensors, semiconductor thin film device fabrication and characterisation.\nAt the moment I have very strong interest in radiation environmental pollution and bacteriology treatment. The teams of researchers are working very hard to bring novel results in this field. I am also a member of the team in charge for the supervision of Ph.D. students in the fields of development of silicon based planar waveguide sensor devices, study of inelastic electron tunnelling in planar tunnelling nanostructures for sensing applications and development of organotellurium(IV) compounds for semiconductor applications. I am a specialist in data analysis techniques and nanosurface structure. I have served as the editor for many books, been a member of the editorial board in science journals, have published many papers and hold many patents.",institutionString:null,institution:{name:"Sheffield Hallam University",country:{name:"United Kingdom"}}},{id:"12392",title:"Mr.",name:"Alex",middleName:null,surname:"Lazinica",slug:"alex-lazinica",fullName:"Alex Lazinica",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/12392/images/7282_n.png",biography:"Alex Lazinica is the founder and CEO of IntechOpen. After obtaining a Master's degree in Mechanical Engineering, he continued his PhD studies in Robotics at the Vienna University of Technology. Here he worked as a robotic researcher with the university's Intelligent Manufacturing Systems Group as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and most importantly he co-founded and built the International Journal of Advanced Robotic Systems- world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career, since it was a pathway to founding IntechOpen - Open Access publisher focused on addressing academic researchers needs. Alex is a personification of IntechOpen key values being trusted, open and entrepreneurial. Today his focus is on defining the growth and development strategy for the company.",institutionString:null,institution:{name:"TU Wien",country:{name:"Austria"}}},{id:"62389",title:"PhD.",name:"Ali Demir",middleName:null,surname:"Sezer",slug:"ali-demir-sezer",fullName:"Ali Demir Sezer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/62389/images/3413_n.jpg",biography:"Dr. Ali Demir Sezer has a Ph.D. from Pharmaceutical Biotechnology at the Faculty of Pharmacy, University of Marmara (Turkey). He is the member of many Pharmaceutical Associations and acts as a reviewer of scientific journals and European projects under different research areas such as: drug delivery systems, nanotechnology and pharmaceutical biotechnology. Dr. Sezer is the author of many scientific publications in peer-reviewed journals and poster communications. Focus of his research activity is drug delivery, physico-chemical characterization and biological evaluation of biopolymers micro and nanoparticles as modified drug delivery system, and colloidal drug carriers (liposomes, nanoparticles etc.).",institutionString:null,institution:{name:"Marmara University",country:{name:"Turkey"}}},{id:"64434",title:"Dr.",name:"Angkoon",middleName:null,surname:"Phinyomark",slug:"angkoon-phinyomark",fullName:"Angkoon Phinyomark",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/64434/images/2619_n.jpg",biography:"My name is Angkoon Phinyomark. I received a B.Eng. degree in Computer Engineering with First Class Honors in 2008 from Prince of Songkla University, Songkhla, Thailand, where I received a Ph.D. degree in Electrical Engineering. My research interests are primarily in the area of biomedical signal processing and classification notably EMG (electromyography signal), EOG (electrooculography signal), and EEG (electroencephalography signal), image analysis notably breast cancer analysis and optical coherence tomography, and rehabilitation engineering. I became a student member of IEEE in 2008. During October 2011-March 2012, I had worked at School of Computer Science and Electronic Engineering, University of Essex, Colchester, Essex, United Kingdom. In addition, during a B.Eng. I had been a visiting research student at Faculty of Computer Science, University of Murcia, Murcia, Spain for three months.\n\nI have published over 40 papers during 5 years in refereed journals, books, and conference proceedings in the areas of electro-physiological signals processing and classification, notably EMG and EOG signals, fractal analysis, wavelet analysis, texture analysis, feature extraction and machine learning algorithms, and assistive and rehabilitative devices. I have several computer programming language certificates, i.e. Sun Certified Programmer for the Java 2 Platform 1.4 (SCJP), Microsoft Certified Professional Developer, Web Developer (MCPD), Microsoft Certified Technology Specialist, .NET Framework 2.0 Web (MCTS). I am a Reviewer for several refereed journals and international conferences, such as IEEE Transactions on Biomedical Engineering, IEEE Transactions on Industrial Electronics, Optic Letters, Measurement Science Review, and also a member of the International Advisory Committee for 2012 IEEE Business Engineering and Industrial Applications and 2012 IEEE Symposium on Business, Engineering and Industrial Applications.",institutionString:null,institution:{name:"Joseph Fourier University",country:{name:"France"}}},{id:"55578",title:"Dr.",name:"Antonio",middleName:null,surname:"Jurado-Navas",slug:"antonio-jurado-navas",fullName:"Antonio Jurado-Navas",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/55578/images/4574_n.png",biography:"Antonio Jurado-Navas received the M.S. degree (2002) and the Ph.D. degree (2009) in Telecommunication Engineering, both from the University of Málaga (Spain). He first worked as a consultant at Vodafone-Spain. From 2004 to 2011, he was a Research Assistant with the Communications Engineering Department at the University of Málaga. In 2011, he became an Assistant Professor in the same department. From 2012 to 2015, he was with Ericsson Spain, where he was working on geo-location\ntools for third generation mobile networks. Since 2015, he is a Marie-Curie fellow at the Denmark Technical University. His current research interests include the areas of mobile communication systems and channel modeling in addition to atmospheric optical communications, adaptive optics and statistics",institutionString:null,institution:{name:"University of Malaga",country:{name:"Spain"}}},{id:"6495",title:"Dr.",name:"Daniel",middleName:null,surname:"Eberli",slug:"daniel-eberli",fullName:"Daniel Eberli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/6495/images/1947_n.jpg",biography:"Daniel Eberli MD. Ph.D. is a scientific physician working in the translational field of urologic tissue engineering. He has a medical degree from the Medical School in Zurich, Switzerland, and a Ph.D. in Molecular Medicine from Wake Forest University, Winston Salem, NC. He currently has a faculty position at the Department of Urology at the University Hospital Zurich, where he devotes half of his time to patient care. He is a lecturer at the Medical School of Zurich and the Swiss Federal Institute of Technology. Together with his research team, he is working on novel biomaterials for bladder reconstruction, improving autonomic innervation, cellular treatment of incontinence and tracking of stem cells.",institutionString:null,institution:{name:"University Hospital of Zurich",country:{name:"Switzerland"}}},{id:"122240",title:"Prof.",name:"Frede",middleName:null,surname:"Blaabjerg",slug:"frede-blaabjerg",fullName:"Frede Blaabjerg",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Aalborg University",country:{name:"Denmark"}}},{id:"50823",title:"Prof.",name:"Hamid Reza",middleName:null,surname:"Karimi",slug:"hamid-reza-karimi",fullName:"Hamid Reza Karimi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Polytechnic University of Milan",country:{name:"Italy"}}},{id:"22128",title:"Dr.",name:"Harald",middleName:null,surname:"Haas",slug:"harald-haas",fullName:"Harald Haas",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Edinburgh",country:{name:"United Kingdom"}}},{id:"80399",title:"Dr.",name:"Huosheng",middleName:null,surname:"Hu",slug:"huosheng-hu",fullName:"Huosheng Hu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Essex",country:{name:"United Kingdom"}}},{id:"107729",title:"Prof.",name:"Joachim",middleName:null,surname:"Maier",slug:"joachim-maier",fullName:"Joachim Maier",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Max Planck Institute for Solid State Research",country:{name:"Germany"}}},{id:"51995",title:"Dr.",name:"Juan A.",middleName:"A.",surname:"Blanco",slug:"juan-a.-blanco",fullName:"Juan A. Blanco",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/51995/images/1076_n.jpg",biography:"Dr. Blanco is an Assistant Professor at the Public University of Navarre. His work is focused on the development and evaluation of ecological models to simulate the influences of management, climate and other ecological factors on tree growth. He is currently collaborating with research teams from Canada, Taiwan, USA, Spain, Cuba, and China in using ecological models to explore the effects of climate change, atmospheric pollution and alternative forest practices in natural and planted forest in boreal, temperate and tropical forests. His research has been applied in mining to optimize reclamation plans, in forestry to assess the potential for carbon sequestration and by government agencies to define local guidelines for long-term sustainable forest management. Among other topics related to forest ecology, Dr. Blanco has studied the influence of climate variations on tree growth and estimated the possible ecological consequences of climate change in forest ecosystems. He has also co-authored the first book dedicated exclusively to the use of hybrid ecological models in forest management, entitled “Forecasting Forest Futures” (Earthscan, London), edited three books on Climate Change effects, mitigation and adaptation (InTech, Rijeka), and three more on Forest Ecosystems, Biodiversity and Tropical Forests (InTech, Rijeka).",institutionString:null,institution:{name:"Universidad Publica De Navarra",country:{name:"Spain"}}}],filtersByRegion:[{group:"region",caption:"North America",value:1,count:5311},{group:"region",caption:"Middle and South America",value:2,count:4814},{group:"region",caption:"Africa",value:3,count:1465},{group:"region",caption:"Asia",value:4,count:9355},{group:"region",caption:"Australia and Oceania",value:5,count:837},{group:"region",caption:"Europe",value:6,count:14773}],offset:12,limit:12,total:14778},chapterEmbeded:{data:{}},editorApplication:{success:null,errors:{}},ofsBooks:{filterParams:{sort:"dateEndSecondStepPublish"},books:[{type:"book",id:"7852",title:"Binding Immunoglobulin Protein",subtitle:null,isOpenForSubmission:!0,hash:"2e2e79aa033b6f1c096eb1fb9be03d1c",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/7852.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7901",title:"Cocoa",subtitle:null,isOpenForSubmission:!0,hash:"bd93f97ceb11fd901da97e54a700270d",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/7901.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8196",title:"ABC Transporters",subtitle:null,isOpenForSubmission:!0,hash:"bfec00d7a6a9666fe01c230f7b133297",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8196.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8367",title:"Collagen",subtitle:null,isOpenForSubmission:!0,hash:"cebbd18b88d65288e3b0d4d0d1050830",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8367.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8565",title:"Aeronautics and Astronautics",subtitle:null,isOpenForSubmission:!0,hash:"43f114ba03e5e42ba53da372ffc3cbde",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8565.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8568",title:"Intelligent Life Beyond Earth",subtitle:null,isOpenForSubmission:!0,hash:"f678594f3e4c852a12790aca46dbcdc2",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8568.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8601",title:"Fixed Point",subtitle:null,isOpenForSubmission:!0,hash:"294c402de44bdd6636c0bc0ee84333ff",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8601.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8657",title:"Fingerprint Identification System",subtitle:null,isOpenForSubmission:!0,hash:"282a558df729691eb1f767bd57fa3303",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8657.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8733",title:"Side-effects of Penicilin",subtitle:null,isOpenForSubmission:!0,hash:"bcb5e7b509de6ba98f7ed4eeff208df0",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8733.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8764",title:"Methanol Chemistry",subtitle:null,isOpenForSubmission:!0,hash:"a2d585554682fa7c2ad74c79ceb2cfee",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8764.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8682",title:"Vectors",subtitle:null,isOpenForSubmission:!0,hash:"a9ea6743964f79b391b672fc667bbbf1",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8682.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8696",title:"Pulmonary Insufficiency",subtitle:null,isOpenForSubmission:!0,hash:"0dfed0b2495fdf08ebf2f5779db08135",slug:null,bookSignature:"",coverURL:"https://cdn.intechopen.com/books/images_new/8696.jpg",editedByType:null,editors:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],filtersByTopic:[{group:"topic",caption:"Agricultural and Biological Sciences",value:5,count:35},{group:"topic",caption:"Biochemistry, Genetics and Molecular Biology",value:6,count:33},{group:"topic",caption:"Business, Management and Economics",value:7,count:9},{group:"topic",caption:"Chemistry",value:8,count:28},{group:"topic",caption:"Computer and Information Science",value:9,count:27},{group:"topic",caption:"Earth and Planetary Sciences",value:10,count:15},{group:"topic",caption:"Engineering",value:11,count:75},{group:"topic",caption:"Environmental Sciences",value:12,count:13},{group:"topic",caption:"Immunology and Microbiology",value:13,count:3},{group:"topic",caption:"Materials Science",value:14,count:39},{group:"topic",caption:"Mathematics",value:15,count:14},{group:"topic",caption:"Medicine",value:16,count:142},{group:"topic",caption:"Nanotechnology and Nanomaterials",value:17,count:7},{group:"topic",caption:"Neuroscience",value:18,count:6},{group:"topic",caption:"Pharmacology, Toxicology and Pharmaceutical Science",value:19,count:7},{group:"topic",caption:"Physics",value:20,count:25},{group:"topic",caption:"Psychology",value:21,count:2},{group:"topic",caption:"Robotics",value:22,count:6},{group:"topic",caption:"Social Sciences",value:23,count:14},{group:"topic",caption:"Technology",value:24,count:12},{group:"topic",caption:"Veterinary Medicine and Science",value:25,count:3},{group:"topic",caption:"Intelligent System",value:535,count:1},{group:"topic",caption:"Osteology",value:1414,count:1},{group:"topic",caption:"Polymer Chemistry",value:1415,count:1}],offset:12,limit:12,total:998},popularBooks:{featuredBooks:[{type:"book",id:"7878",title:"Advances in Extracorporeal Membrane Oxygenation",subtitle:"Volume 3",isOpenForSubmission:!1,hash:"f95bf990273d08098a00f9a1c2403cbe",slug:"advances-in-extracorporeal-membrane-oxygenation-volume-3",bookSignature:"Michael S. Firstenberg",coverURL:"https://cdn.intechopen.com/books/images_new/7878.jpg",editors:[{id:"64343",title:null,name:"Michael S.",middleName:"S",surname:"Firstenberg",slug:"michael-s.-firstenberg",fullName:"Michael S. Firstenberg"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7614",title:"Fourier Transforms",subtitle:"Century of Digitalization and Increasing Expectations",isOpenForSubmission:!1,hash:"ff3501657ae983a3b42fef1f7058ac91",slug:"fourier-transforms-century-of-digitalization-and-increasing-expectations",bookSignature:"Goran S. Nikoli? and Dragana Z. Markovi?-Nikoli?",coverURL:"https://cdn.intechopen.com/books/images_new/7614.jpg",editors:[{id:"23261",title:"Prof.",name:"Goran",middleName:"S.",surname:"Nikolic",slug:"goran-nikolic",fullName:"Goran Nikolic"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8299",title:"Timber Buildings and Sustainability",subtitle:null,isOpenForSubmission:!1,hash:"bccf2891cec38ed041724131aa34c25a",slug:"timber-buildings-and-sustainability",bookSignature:"Giovanna Concu",coverURL:"https://cdn.intechopen.com/books/images_new/8299.jpg",editors:[{id:"108709",title:"Dr.",name:"Giovanna",middleName:null,surname:"Concu",slug:"giovanna-concu",fullName:"Giovanna Concu"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7062",title:"Rhinosinusitis",subtitle:null,isOpenForSubmission:!1,hash:"14ed95e155b1e57a61827ca30b579d09",slug:"rhinosinusitis",bookSignature:"Balwant Singh Gendeh and Mirjana Turkalj",coverURL:"https://cdn.intechopen.com/books/images_new/7062.jpg",editors:[{id:"67669",title:"Prof.",name:"Balwant Singh",middleName:null,surname:"Gendeh",slug:"balwant-singh-gendeh",fullName:"Balwant Singh Gendeh"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7087",title:"Tendons",subtitle:null,isOpenForSubmission:!1,hash:"786abac0445c102d1399a1e727a2db7f",slug:"tendons",bookSignature:"Hasan Sözen",coverURL:"https://cdn.intechopen.com/books/images_new/7087.jpg",editors:[{id:"161402",title:"Dr.",name:"Hasan",middleName:null,surname:"Sözen",slug:"hasan-sozen",fullName:"Hasan Sözen"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7955",title:"Advances in Hematologic Malignancies",subtitle:null,isOpenForSubmission:!1,hash:"59ca1b09447fab4717a93e099f646d28",slug:"advances-in-hematologic-malignancies",bookSignature:"Gamal Abdul Hamid",coverURL:"https://cdn.intechopen.com/books/images_new/7955.jpg",editors:[{id:"36487",title:"Prof.",name:"Gamal",middleName:null,surname:"Abdul Hamid",slug:"gamal-abdul-hamid",fullName:"Gamal Abdul Hamid"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7701",title:"Assistive and Rehabilitation Engineering",subtitle:null,isOpenForSubmission:!1,hash:"4191b744b8af3b17d9a80026dcb0617f",slug:"assistive-and-rehabilitation-engineering",bookSignature:"Yves Rybarczyk",coverURL:"https://cdn.intechopen.com/books/images_new/7701.jpg",editors:[{id:"72920",title:"Prof.",name:"Yves",middleName:"Philippe",surname:"Rybarczyk",slug:"yves-rybarczyk",fullName:"Yves Rybarczyk"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7837",title:"Geriatric Medicine and Gerontology",subtitle:null,isOpenForSubmission:!1,hash:"e277d005b23536bcd9f8550046101979",slug:"geriatric-medicine-and-gerontology",bookSignature:"Edward T. Zawada Jr.",coverURL:"https://cdn.intechopen.com/books/images_new/7837.jpg",editors:[{id:"16344",title:"Dr.",name:"Edward T.",middleName:null,surname:"Zawada Jr.",slug:"edward-t.-zawada-jr.",fullName:"Edward T. Zawada Jr."}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7123",title:"Current Topics in Neglected Tropical Diseases",subtitle:null,isOpenForSubmission:!1,hash:"61c627da05b2ace83056d11357bdf361",slug:"current-topics-in-neglected-tropical-diseases",bookSignature:"Alfonso J. Rodriguez-Morales",coverURL:"https://cdn.intechopen.com/books/images_new/7123.jpg",editors:[{id:"131400",title:"Dr.",name:"Alfonso J.",middleName:null,surname:"Rodriguez-Morales",slug:"alfonso-j.-rodriguez-morales",fullName:"Alfonso J. Rodriguez-Morales"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7610",title:"Renewable and Sustainable Composites",subtitle:null,isOpenForSubmission:!1,hash:"c2de26c3d329c54f093dc3f05417500a",slug:"renewable-and-sustainable-composites",bookSignature:"António B. Pereira and Fábio A. O. Fernandes",coverURL:"https://cdn.intechopen.com/books/images_new/7610.jpg",editors:[{id:"211131",title:"Prof.",name:"António",middleName:"Bastos",surname:"Pereira",slug:"antonio-pereira",fullName:"António Pereira"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8416",title:"Non-Equilibrium Particle Dynamics",subtitle:null,isOpenForSubmission:!1,hash:"2c3add7639dcd1cb442cb4313ea64e3a",slug:"non-equilibrium-particle-dynamics",bookSignature:"Albert S. Kim",coverURL:"https://cdn.intechopen.com/books/images_new/8416.jpg",editors:[{id:"21045",title:"Prof.",name:"Albert S.",middleName:null,surname:"Kim",slug:"albert-s.-kim",fullName:"Albert S. Kim"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8463",title:"Pediatric Surgery, Flowcharts and Clinical Algorithms",subtitle:null,isOpenForSubmission:!1,hash:"23f39beea4d557b0ae424e2eaf82bf5e",slug:"pediatric-surgery-flowcharts-and-clinical-algorithms",bookSignature:"Sameh Shehata",coverURL:"https://cdn.intechopen.com/books/images_new/8463.jpg",editors:[{id:"37518",title:"Prof.",name:"Sameh",middleName:null,surname:"Shehata",slug:"sameh-shehata",fullName:"Sameh Shehata"}],productType:{id:"1",chapterContentType:"chapter"}}],offset:12,limit:12,total:4385},hotBookTopics:{hotBooks:[],offset:0,limit:12,total:null},publish:{},publishingProposal:{success:null,errors:{}},books:{featuredBooks:[{type:"book",id:"7878",title:"Advances in Extracorporeal Membrane Oxygenation",subtitle:"Volume 3",isOpenForSubmission:!1,hash:"f95bf990273d08098a00f9a1c2403cbe",slug:"advances-in-extracorporeal-membrane-oxygenation-volume-3",bookSignature:"Michael S. Firstenberg",coverURL:"https://cdn.intechopen.com/books/images_new/7878.jpg",editors:[{id:"64343",title:null,name:"Michael S.",middleName:"S",surname:"Firstenberg",slug:"michael-s.-firstenberg",fullName:"Michael S. Firstenberg"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7614",title:"Fourier Transforms",subtitle:"Century of Digitalization and Increasing Expectations",isOpenForSubmission:!1,hash:"ff3501657ae983a3b42fef1f7058ac91",slug:"fourier-transforms-century-of-digitalization-and-increasing-expectations",bookSignature:"Goran S. Nikoli? and Dragana Z. Markovi?-Nikoli?",coverURL:"https://cdn.intechopen.com/books/images_new/7614.jpg",editors:[{id:"23261",title:"Prof.",name:"Goran",middleName:"S.",surname:"Nikolic",slug:"goran-nikolic",fullName:"Goran Nikolic"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8299",title:"Timber Buildings and Sustainability",subtitle:null,isOpenForSubmission:!1,hash:"bccf2891cec38ed041724131aa34c25a",slug:"timber-buildings-and-sustainability",bookSignature:"Giovanna Concu",coverURL:"https://cdn.intechopen.com/books/images_new/8299.jpg",editors:[{id:"108709",title:"Dr.",name:"Giovanna",middleName:null,surname:"Concu",slug:"giovanna-concu",fullName:"Giovanna Concu"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7062",title:"Rhinosinusitis",subtitle:null,isOpenForSubmission:!1,hash:"14ed95e155b1e57a61827ca30b579d09",slug:"rhinosinusitis",bookSignature:"Balwant Singh Gendeh and Mirjana Turkalj",coverURL:"https://cdn.intechopen.com/books/images_new/7062.jpg",editors:[{id:"67669",title:"Prof.",name:"Balwant Singh",middleName:null,surname:"Gendeh",slug:"balwant-singh-gendeh",fullName:"Balwant Singh Gendeh"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7087",title:"Tendons",subtitle:null,isOpenForSubmission:!1,hash:"786abac0445c102d1399a1e727a2db7f",slug:"tendons",bookSignature:"Hasan Sözen",coverURL:"https://cdn.intechopen.com/books/images_new/7087.jpg",editors:[{id:"161402",title:"Dr.",name:"Hasan",middleName:null,surname:"Sözen",slug:"hasan-sozen",fullName:"Hasan Sözen"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7955",title:"Advances in Hematologic Malignancies",subtitle:null,isOpenForSubmission:!1,hash:"59ca1b09447fab4717a93e099f646d28",slug:"advances-in-hematologic-malignancies",bookSignature:"Gamal Abdul Hamid",coverURL:"https://cdn.intechopen.com/books/images_new/7955.jpg",editors:[{id:"36487",title:"Prof.",name:"Gamal",middleName:null,surname:"Abdul Hamid",slug:"gamal-abdul-hamid",fullName:"Gamal Abdul Hamid"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7701",title:"Assistive and Rehabilitation Engineering",subtitle:null,isOpenForSubmission:!1,hash:"4191b744b8af3b17d9a80026dcb0617f",slug:"assistive-and-rehabilitation-engineering",bookSignature:"Yves Rybarczyk",coverURL:"https://cdn.intechopen.com/books/images_new/7701.jpg",editors:[{id:"72920",title:"Prof.",name:"Yves",middleName:"Philippe",surname:"Rybarczyk",slug:"yves-rybarczyk",fullName:"Yves Rybarczyk"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7837",title:"Geriatric Medicine and Gerontology",subtitle:null,isOpenForSubmission:!1,hash:"e277d005b23536bcd9f8550046101979",slug:"geriatric-medicine-and-gerontology",bookSignature:"Edward T. Zawada Jr.",coverURL:"https://cdn.intechopen.com/books/images_new/7837.jpg",editors:[{id:"16344",title:"Dr.",name:"Edward T.",middleName:null,surname:"Zawada Jr.",slug:"edward-t.-zawada-jr.",fullName:"Edward T. Zawada Jr."}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7123",title:"Current Topics in Neglected Tropical Diseases",subtitle:null,isOpenForSubmission:!1,hash:"61c627da05b2ace83056d11357bdf361",slug:"current-topics-in-neglected-tropical-diseases",bookSignature:"Alfonso J. Rodriguez-Morales",coverURL:"https://cdn.intechopen.com/books/images_new/7123.jpg",editors:[{id:"131400",title:"Dr.",name:"Alfonso J.",middleName:null,surname:"Rodriguez-Morales",slug:"alfonso-j.-rodriguez-morales",fullName:"Alfonso J. Rodriguez-Morales"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"7610",title:"Renewable and Sustainable Composites",subtitle:null,isOpenForSubmission:!1,hash:"c2de26c3d329c54f093dc3f05417500a",slug:"renewable-and-sustainable-composites",bookSignature:"António B. Pereira and Fábio A. O. Fernandes",coverURL:"https://cdn.intechopen.com/books/images_new/7610.jpg",editors:[{id:"211131",title:"Prof.",name:"António",middleName:"Bastos",surname:"Pereira",slug:"antonio-pereira",fullName:"António Pereira"}],productType:{id:"1",chapterContentType:"chapter"}}],latestBooks:[{type:"book",id:"8463",title:"Pediatric Surgery, Flowcharts and Clinical Algorithms",subtitle:null,isOpenForSubmission:!1,hash:"23f39beea4d557b0ae424e2eaf82bf5e",slug:"pediatric-surgery-flowcharts-and-clinical-algorithms",bookSignature:"Sameh Shehata",coverURL:"https://cdn.intechopen.com/books/images_new/8463.jpg",editedByType:"Edited by",editors:[{id:"37518",title:"Prof.",name:"Sameh",middleName:null,surname:"Shehata",slug:"sameh-shehata",fullName:"Sameh Shehata"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7187",title:"Osteosarcoma",subtitle:"Diagnosis, Mechanisms, and Translational Developments",isOpenForSubmission:!1,hash:"89096359b754beb806eca4c6d8aacaba",slug:"osteosarcoma-diagnosis-mechanisms-and-translational-developments",bookSignature:"Matthew Gregory Cable and Robert Lawrence Randall",coverURL:"https://cdn.intechopen.com/books/images_new/7187.jpg",editedByType:"Edited by",editors:[{id:"265693",title:"Dr.",name:"Matthew Gregory",middleName:null,surname:"Cable",slug:"matthew-gregory-cable",fullName:"Matthew Gregory Cable"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7955",title:"Advances in Hematologic Malignancies",subtitle:null,isOpenForSubmission:!1,hash:"59ca1b09447fab4717a93e099f646d28",slug:"advances-in-hematologic-malignancies",bookSignature:"Gamal Abdul Hamid",coverURL:"https://cdn.intechopen.com/books/images_new/7955.jpg",editedByType:"Edited by",editors:[{id:"36487",title:"Prof.",name:"Gamal",middleName:null,surname:"Abdul Hamid",slug:"gamal-abdul-hamid",fullName:"Gamal Abdul Hamid"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7701",title:"Assistive and Rehabilitation Engineering",subtitle:null,isOpenForSubmission:!1,hash:"4191b744b8af3b17d9a80026dcb0617f",slug:"assistive-and-rehabilitation-engineering",bookSignature:"Yves Rybarczyk",coverURL:"https://cdn.intechopen.com/books/images_new/7701.jpg",editedByType:"Edited by",editors:[{id:"72920",title:"Prof.",name:"Yves",middleName:"Philippe",surname:"Rybarczyk",slug:"yves-rybarczyk",fullName:"Yves Rybarczyk"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7726",title:"Swarm Intelligence",subtitle:"Recent Advances, New Perspectives and Applications",isOpenForSubmission:!1,hash:"e7ea7e74ce7a7a8e5359629e07c68d31",slug:"swarm-intelligence-recent-advances-new-perspectives-and-applications",bookSignature:"Javier Del Ser, Esther Villar and Eneko Osaba",coverURL:"https://cdn.intechopen.com/books/images_new/7726.jpg",editedByType:"Edited by",editors:[{id:"49813",title:"Dr.",name:"Javier",middleName:null,surname:"Del Ser",slug:"javier-del-ser",fullName:"Javier Del Ser"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8256",title:"Distillation",subtitle:"Modelling, Simulation and Optimization",isOpenForSubmission:!1,hash:"c76af109f83e14d915e5cb3949ae8b80",slug:"distillation-modelling-simulation-and-optimization",bookSignature:"Vilmar Steffen",coverURL:"https://cdn.intechopen.com/books/images_new/8256.jpg",editedByType:"Edited by",editors:[{id:"189035",title:"Dr.",name:"Vilmar",middleName:null,surname:"Steffen",slug:"vilmar-steffen",fullName:"Vilmar Steffen"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7240",title:"Growing and Handling of Bacterial Cultures",subtitle:null,isOpenForSubmission:!1,hash:"a76c3ef7718c0b72d0128817cdcbe6e3",slug:"growing-and-handling-of-bacterial-cultures",bookSignature:"Madhusmita Mishra",coverURL:"https://cdn.intechopen.com/books/images_new/7240.jpg",editedByType:"Edited by",editors:[{id:"204267",title:"Dr.",name:"Madhusmita",middleName:null,surname:"Mishra",slug:"madhusmita-mishra",fullName:"Madhusmita Mishra"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8299",title:"Timber Buildings and Sustainability",subtitle:null,isOpenForSubmission:!1,hash:"bccf2891cec38ed041724131aa34c25a",slug:"timber-buildings-and-sustainability",bookSignature:"Giovanna Concu",coverURL:"https://cdn.intechopen.com/books/images_new/8299.jpg",editedByType:"Edited by",editors:[{id:"108709",title:"Dr.",name:"Giovanna",middleName:null,surname:"Concu",slug:"giovanna-concu",fullName:"Giovanna Concu"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7062",title:"Rhinosinusitis",subtitle:null,isOpenForSubmission:!1,hash:"14ed95e155b1e57a61827ca30b579d09",slug:"rhinosinusitis",bookSignature:"Balwant Singh Gendeh and Mirjana Turkalj",coverURL:"https://cdn.intechopen.com/books/images_new/7062.jpg",editedByType:"Edited by",editors:[{id:"67669",title:"Prof.",name:"Balwant Singh",middleName:null,surname:"Gendeh",slug:"balwant-singh-gendeh",fullName:"Balwant Singh Gendeh"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7837",title:"Geriatric Medicine and Gerontology",subtitle:null,isOpenForSubmission:!1,hash:"e277d005b23536bcd9f8550046101979",slug:"geriatric-medicine-and-gerontology",bookSignature:"Edward T. Zawada Jr.",coverURL:"https://cdn.intechopen.com/books/images_new/7837.jpg",editedByType:"Edited by",editors:[{id:"16344",title:"Dr.",name:"Edward T.",middleName:null,surname:"Zawada Jr.",slug:"edward-t.-zawada-jr.",fullName:"Edward T. Zawada Jr."}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},subject:{topic:{id:"875",title:"Ecosystem",slug:"environmental-sciences-forestry-science-ecosystem",parent:{title:"Forestry Science",slug:"environmental-sciences-forestry-science"},numberOfBooks:2,numberOfAuthorsAndEditors:57,numberOfWosCitations:62,numberOfCrossrefCitations:35,numberOfDimensionsCitations:104,videoUrl:null,fallbackUrl:null,description:null},booksByTopicFilter:{topicSlug:"environmental-sciences-forestry-science-ecosystem",sort:"-publishedDate",limit:12,offset:0},booksByTopicCollection:[{type:"book",id:"5539",title:"Forest Ecology and Conservation",subtitle:null,isOpenForSubmission:!1,hash:"6bd160f6d1da73fc253dfe6c4df7c095",slug:"forest-ecology-and-conservation",bookSignature:"Sumit Chakravarty and Gopal Shukla",coverURL:"https://cdn.intechopen.com/books/images_new/5539.jpg",editedByType:"Edited by",editors:[{id:"101105",title:"Dr.",name:"Gopal",middleName:null,surname:"Shukla",slug:"gopal-shukla",fullName:"Gopal Shukla"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"902",title:"Tropical Forests",subtitle:null,isOpenForSubmission:!1,hash:"55286837c680e9be2bc357abf678212e",slug:"tropical-forests",bookSignature:"Padmini Sudarshana, Madhugiri Nageswara-Rao and Jaya R. Soneji",coverURL:"https://cdn.intechopen.com/books/images_new/902.jpg",editedByType:"Edited by",editors:[{id:"79318",title:"Dr.",name:"Padmini",middleName:null,surname:"Sudarshana",slug:"padmini-sudarshana",fullName:"Padmini Sudarshana"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],booksByTopicTotal:2,mostCitedChapters:[{id:"31970",doi:"10.5772/33085",title:"A Review of Above Ground Necromass in Tropical Forests",slug:"a-review-of-above-ground-necromass-in-tropical-forests",totalDownloads:2501,totalCrossrefCites:9,totalDimensionsCites:25,book:{slug:"tropical-forests",title:"Tropical Forests",fullTitle:"Tropical Forests"},signatures:"Michael Palace, Michael Keller, George Hurtt and Steve Frolking",authors:[{id:"93962",title:"Dr.",name:"Michael",middleName:null,surname:"Palace",slug:"michael-palace",fullName:"Michael Palace"},{id:"93987",title:"Dr.",name:"Michael",middleName:null,surname:"Keller",slug:"michael-keller",fullName:"Michael Keller"},{id:"93989",title:"Dr.",name:"Steve",middleName:null,surname:"Frolking",slug:"steve-frolking",fullName:"Steve Frolking"},{id:"93992",title:"Dr.",name:"George",middleName:null,surname:"Hurtt",slug:"george-hurtt",fullName:"George Hurtt"}]},{id:"31972",doi:"10.5772/31523",title:"Measuring Tropical Deforestation with Error Margins: A Method for REDD Monitoring in South-Eastern Mexico",slug:"measuring-tropical-deforestation-with-error-margins-a-method-for-redd-monitoring-in-south-eastern-me",totalDownloads:2458,totalCrossrefCites:2,totalDimensionsCites:21,book:{slug:"tropical-forests",title:"Tropical Forests",fullTitle:"Tropical Forests"},signatures:"Stéphane Couturier, Juan Manuel Núñez and Melanie Kolb",authors:[{id:"87605",title:"Dr",name:"Stéphane",middleName:null,surname:"Couturier",slug:"stephane-couturier",fullName:"Stéphane Couturier"},{id:"93234",title:"MSc.",name:"Juan",middleName:null,surname:"Nunez",slug:"juan-nunez",fullName:"Juan Nunez"},{id:"145885",title:"MSc.",name:"Melanie",middleName:null,surname:"Kolb",slug:"melanie-kolb",fullName:"Melanie Kolb"}]},{id:"31964",doi:"10.5772/33217",title:"Dispersion, an Important Radiation Mechanism for Ectomycorrhizal Fungi in Neotropical Lowland Forests?",slug:"dispersion-an-important-radiation-mechanism-for-ectomycorrhizal-fungi-in-neotropical-lowland-forests",totalDownloads:2654,totalCrossrefCites:9,totalDimensionsCites:11,book:{slug:"tropical-forests",title:"Tropical Forests",fullTitle:"Tropical Forests"},signatures:"Bernard Moyersoen",authors:[{id:"94510",title:"Dr.",name:"Bernard",middleName:null,surname:"Moyersoen",slug:"bernard-moyersoen",fullName:"Bernard Moyersoen"}]}],mostDownloadedChaptersLast30Days:[{id:"66710",title:"Deforestation in India: Consequences and Sustainable Solutions",slug:"deforestation-in-india-consequences-and-sustainable-solutions",totalDownloads:149,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:null,title:"Deforestation Around the World",fullTitle:"Deforestation Around the World"},signatures:"Rima Kumari, Ayan Banerjee, Rahul Kumar, Amit Kumar, Purabi Saikia and Mohammed Latif Khan",authors:null},{id:"68528",title:"Forest Biodiversity and Deforestation in Bangladesh: The Latest Update",slug:"forest-biodiversity-and-deforestation-in-bangladesh-the-latest-update",totalDownloads:38,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:null,title:"Deforestation Around the World",fullTitle:"Deforestation Around the World"},signatures:"Ahm Ali Reza and Md. Kamrul Hasan",authors:null},{id:"31971",title:"Seasonal Pattern of Vegetative Cover from NDVI Time-Series",slug:"seasonal-pattern-of-vegetative-cover-from-ndvi-time-series-",totalDownloads:3711,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:"tropical-forests",title:"Tropical Forests",fullTitle:"Tropical Forests"},signatures:"Dyah R. Panuju and Bambang H. Trisasongko",authors:[{id:"82110",title:"Ms.",name:"Dyah",middleName:"Retno",surname:"Panuju",slug:"dyah-panuju",fullName:"Dyah Panuju"},{id:"88822",title:"Mr.",name:"Bambang",middleName:null,surname:"Trisasongko",slug:"bambang-trisasongko",fullName:"Bambang Trisasongko"}]},{id:"54603",title:"Methodological Considerations in the Study of Earthworms in Forest Ecosystems",slug:"methodological-considerations-in-the-study-of-earthworms-in-forest-ecosystems",totalDownloads:906,totalCrossrefCites:0,totalDimensionsCites:1,book:{slug:"forest-ecology-and-conservation",title:"Forest Ecology and Conservation",fullTitle:"Forest Ecology and Conservation"},signatures:"Dylan Rhea-Fournier and Grizelle González",authors:[{id:"82355",title:"Dr.",name:"Grizelle",middleName:null,surname:"Gonzalez",slug:"grizelle-gonzalez",fullName:"Grizelle Gonzalez"},{id:"194800",title:"M.Sc.",name:"Dylan",middleName:null,surname:"Rhea-Fournier",slug:"dylan-rhea-fournier",fullName:"Dylan Rhea-Fournier"}]},{id:"31959",title:"Structure, Diversity, Threats and Conservation of Tropical Forests",slug:"structure-diversity-threats-and-conservation-of-tropical-forests",totalDownloads:6869,totalCrossrefCites:1,totalDimensionsCites:2,book:{slug:"tropical-forests",title:"Tropical Forests",fullTitle:"Tropical Forests"},signatures:"Madhugiri Nageswara-Rao, Jaya R. Soneji and Padmini Sudarshana",authors:[{id:"79318",title:"Dr.",name:"Padmini",middleName:null,surname:"Sudarshana",slug:"padmini-sudarshana",fullName:"Padmini Sudarshana"},{id:"120847",title:"Dr.",name:"Madhugiri",middleName:null,surname:"Nageswara-Rao",slug:"madhugiri-nageswara-rao",fullName:"Madhugiri Nageswara-Rao"},{id:"120848",title:"Dr.",name:"Jaya",middleName:null,surname:"Soneji",slug:"jaya-soneji",fullName:"Jaya Soneji"}]},{id:"31969",title:"Floristic Composition, Diversity and Status of Threatened Medicinal Plants in Tropical Forests of Malyagiri Hill Ranges, Eastern Ghats, India",slug:"floristic-composition-diversity-and-status-of-threatened-medicinal-plants-in-tropical-forests-of-mal",totalDownloads:3399,totalCrossrefCites:0,totalDimensionsCites:1,book:{slug:"tropical-forests",title:"Tropical Forests",fullTitle:"Tropical Forests"},signatures:"S. C. Sahu and N. K. Dhal",authors:[{id:"78313",title:"Dr.",name:"Nabin Kumar",middleName:null,surname:"Dhal",slug:"nabin-kumar-dhal",fullName:"Nabin Kumar Dhal"},{id:"89118",title:"Dr.",name:"Sudam Charan",middleName:null,surname:"Sahu",slug:"sudam-charan-sahu",fullName:"Sudam Charan Sahu"}]},{id:"67213",title:"Colombian Forest Monitoring System: Assessing Deforestation in an Environmental Complex Country",slug:"colombian-forest-monitoring-system-assessing-deforestation-in-an-environmental-complex-country",totalDownloads:111,totalCrossrefCites:0,totalDimensionsCites:0,book:{slug:null,title:"Deforestation Around the World",fullTitle:"Deforestation Around the World"},signatures:"Edersson Cabrera, Gustavo Galindo, Jose González, Lina Vergara, Cristhian Forero, Alexander Cubillos, Javier Espejo, Juan Rubiano, Xavier Corredor, Leonardo Hurtado, Diana Vargas and Alvaro Duque",authors:null},{id:"66632",title:"Estimating Aboveground Biomass Loss from Deforestation in the Savanna and Semi-arid Biomes of Brazil between 2007 and 2017",slug:"estimating-aboveground-biomass-loss-from-deforestation-in-the-savanna-and-semi-arid-biomes-of-brazil",totalDownloads:207,totalCrossrefCites:0,totalDimensionsCites:2,book:{slug:null,title:"Deforestation Around the World",fullTitle:"Deforestation Around the World"},signatures:"Eduarda Martiniano de Oliveira Silveira, Marcela Castro Nunes Santos Terra, Fausto Weimar Acerbi-Júnior and José Roberto Soares Scolforo",authors:null},{id:"31972",title:"Measuring Tropical Deforestation with Error Margins: A Method for REDD Monitoring in South-Eastern Mexico",slug:"measuring-tropical-deforestation-with-error-margins-a-method-for-redd-monitoring-in-south-eastern-me",totalDownloads:2459,totalCrossrefCites:2,totalDimensionsCites:21,book:{slug:"tropical-forests",title:"Tropical Forests",fullTitle:"Tropical Forests"},signatures:"Stéphane Couturier, Juan Manuel Núñez and Melanie Kolb",authors:[{id:"87605",title:"Dr",name:"Stéphane",middleName:null,surname:"Couturier",slug:"stephane-couturier",fullName:"Stéphane Couturier"},{id:"93234",title:"MSc.",name:"Juan",middleName:null,surname:"Nunez",slug:"juan-nunez",fullName:"Juan Nunez"},{id:"145885",title:"MSc.",name:"Melanie",middleName:null,surname:"Kolb",slug:"melanie-kolb",fullName:"Melanie Kolb"}]},{id:"54299",title:"Remote Sensing and Forest Conservation: Challenges of Illegal Logging in Kursumlija Municipality (Serbia)",slug:"remote-sensing-and-forest-conservation-challenges-of-illegal-logging-in-kursumlija-municipality-serb",totalDownloads:835,totalCrossrefCites:1,totalDimensionsCites:2,book:{slug:"forest-ecology-and-conservation",title:"Forest Ecology and Conservation",fullTitle:"Forest Ecology and Conservation"},signatures:"Miomir M. Jovanović and Miško M. Milanović",authors:[{id:"194569",title:"Prof.",name:"Miomir",middleName:null,surname:"Jovanović",slug:"miomir-jovanovic",fullName:"Miomir Jovanović"},{id:"201206",title:"Dr.",name:"Miško",middleName:null,surname:"Milanović",slug:"misko-milanovic",fullName:"Miško Milanović"}]}],onlineFirstChaptersFilter:{topicSlug:"environmental-sciences-forestry-science-ecosystem",limit:3,offset:0},onlineFirstChaptersCollection:[{id:"68528",title:"Forest Biodiversity and Deforestation in Bangladesh: The Latest Update",slug:"forest-biodiversity-and-deforestation-in-bangladesh-the-latest-update",totalDownloads:38,totalDimensionsCites:0,doi:"10.5772/intechopen.86242",book:{title:"Deforestation Around the World"},signatures:"Ahm Ali Reza and Md. Kamrul Hasan"},{id:"66710",title:"Deforestation in India: Consequences and Sustainable Solutions",slug:"deforestation-in-india-consequences-and-sustainable-solutions",totalDownloads:152,totalDimensionsCites:0,doi:"10.5772/intechopen.85804",book:{title:"Deforestation Around the World"},signatures:"Rima Kumari, Ayan Banerjee, Rahul Kumar, Amit Kumar, Purabi Saikia and Mohammed Latif Khan"},{id:"67213",title:"Colombian Forest Monitoring System: Assessing Deforestation in an Environmental Complex Country",slug:"colombian-forest-monitoring-system-assessing-deforestation-in-an-environmental-complex-country",totalDownloads:112,totalDimensionsCites:0,doi:"10.5772/intechopen.86143",book:{title:"Deforestation Around the World"},signatures:"Edersson Cabrera, Gustavo Galindo, Jose González, Lina Vergara, Cristhian Forero, Alexander Cubillos, Javier Espejo, Juan Rubiano, Xavier Corredor, Leonardo Hurtado, Diana Vargas and Alvaro Duque"}],onlineFirstChaptersTotal:5},preDownload:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[{type:"book",id:"6837",title:"Lithium-ion Batteries - Thin Film for Energy Materials and Devices",subtitle:null,isOpenForSubmission:!0,hash:"ea7789260b319b9a4b472257f57bfeb5",slug:null,bookSignature:"Prof. Mitsunobu Sato, Dr. Li Lu and Dr. Hiroki Nagai",coverURL:"https://cdn.intechopen.com/books/images_new/6837.jpg",editedByType:null,editors:[{id:"179615",title:"Prof.",name:"Mitsunobu",middleName:null,surname:"Sato",slug:"mitsunobu-sato",fullName:"Mitsunobu Sato"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9423",title:"Applications of Artificial Intelligence in Process Industry Automation, Heat and Power Generation and Smart Manufacturing",subtitle:null,isOpenForSubmission:!0,hash:"10ac8fb0bdbf61044395963028653d21",slug:null,bookSignature:"Prof. Konstantinos G. Kyprianidis and Prof. Erik Dahlquist",coverURL:"https://cdn.intechopen.com/books/images_new/9423.jpg",editedByType:null,editors:[{id:"35868",title:"Prof.",name:"Konstantinos",middleName:"G.",surname:"Kyprianidis",slug:"konstantinos-kyprianidis",fullName:"Konstantinos Kyprianidis"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"9428",title:"New Trends in the Use of Artificial Intelligence for the Industry 4.0",subtitle:null,isOpenForSubmission:!0,hash:"9e089eec484ce8e9eb32198c2d8b34ea",slug:null,bookSignature:"Dr. Luis Romeral Martinez, Dr. Roque A. Osornio-Rios and Dr. Miguel Delgado Prieto",coverURL:"https://cdn.intechopen.com/books/images_new/9428.jpg",editedByType:null,editors:[{id:"86501",title:"Dr.",name:"Luis",middleName:null,surname:"Romeral Martinez",slug:"luis-romeral-martinez",fullName:"Luis Romeral Martinez"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10107",title:"Artificial Intelligence in Oncology Drug Discovery & Development",subtitle:null,isOpenForSubmission:!0,hash:"043c178c3668865ab7d35dcb2ceea794",slug:null,bookSignature:"Dr. John Cassidy and Dr. Belle Taylor",coverURL:"https://cdn.intechopen.com/books/images_new/10107.jpg",editedByType:null,editors:[{id:"244455",title:"Dr.",name:"John",middleName:null,surname:"Cassidy",slug:"john-cassidy",fullName:"John Cassidy"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"8903",title:"Carbon Based Material for Environmental Protection and Remediation",subtitle:null,isOpenForSubmission:!0,hash:"19da699b370f320eca63ef2ba02f745d",slug:null,bookSignature:"Dr. Mattia Bartoli and Dr. Marco Frediani",coverURL:"https://cdn.intechopen.com/books/images_new/8903.jpg",editedByType:null,editors:[{id:"188999",title:"Dr.",name:"Mattia",middleName:null,surname:"Bartoli",slug:"mattia-bartoli",fullName:"Mattia Bartoli"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10132",title:"Applied Computational Near-surface Geophysics - From Integral and Derivative Formulas to MATLAB Codes",subtitle:null,isOpenForSubmission:!0,hash:"38cdbbb671df620b36ee96af1d9a3a90",slug:null,bookSignature:"Dr. Afshin Aghayan",coverURL:"https://cdn.intechopen.com/books/images_new/10132.jpg",editedByType:null,editors:[{id:"311030",title:"Dr.",name:"Afshin",middleName:null,surname:"Aghayan",slug:"afshin-aghayan",fullName:"Afshin Aghayan"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10110",title:"Advances and Technologies in Building Construction and Structural Analysis",subtitle:null,isOpenForSubmission:!0,hash:"df2ad14bc5588577e8bf0b7ebcdafd9d",slug:null,bookSignature:"Dr. Ali Kaboli and Dr. Sara Shirowzhan",coverURL:"https://cdn.intechopen.com/books/images_new/10110.jpg",editedByType:null,editors:[{id:"309192",title:"Dr.",name:"Ali",middleName:null,surname:"Kaboli",slug:"ali-kaboli",fullName:"Ali Kaboli"}],productType:{id:"1",chapterContentType:"chapter"}},{type:"book",id:"10175",title:"Ethics in Emerging Technologies",subtitle:null,isOpenForSubmission:!0,hash:"9c92da249676e35e2f7476182aa94e84",slug:null,bookSignature:"Prof. Ali Hessami",coverURL:"https://cdn.intechopen.com/books/images_new/10175.jpg",editedByType:null,editors:[{id:"108303",title:"Prof.",name:"Ali",middleName:null,surname:"Hessami",slug:"ali-hessami",fullName:"Ali Hessami"}],productType:{id:"1",chapterContentType:"chapter"}}],offset:8,limit:8,total:16},humansInSpaceProgram:{},teamHumansInSpaceProgram:{},route:{name:"profile.detail",path:"/profiles/247878/kapil-sharma",hash:"",query:{},params:{id:"247878",slug:"kapil-sharma"},fullPath:"/profiles/247878/kapil-sharma",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)}()