\\n\\n
Dr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\\n\\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\\n\\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\\n\\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\\n\\nThank you all for being part of the journey. 5,000 times thank you!
\\n\\nNow with 5,000 titles available Open Access, which one will you read next?
\\n\\nRead, share and download for free: https://www.intechopen.com/books
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Preparation of Space Experiments edited by international leading expert Dr. Vladimir Pletser, Director of Space Training Operations at Blue Abyss is the 5,000th Open Access book published by IntechOpen and our milestone publication!
\n\n"This book presents some of the current trends in space microgravity research. The eleven chapters introduce various facets of space research in physical sciences, human physiology and technology developed using the microgravity environment not only to improve our fundamental understanding in these domains but also to adapt this new knowledge for application on earth." says the editor. Listen what else Dr. Pletser has to say...
\n\n\n\nDr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\n\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\n\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\n\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\n\nThank you all for being part of the journey. 5,000 times thank you!
\n\nNow with 5,000 titles available Open Access, which one will you read next?
\n\nRead, share and download for free: https://www.intechopen.com/books
\n\n\n\n
\n'}],latestNews:[{slug:"webinar-introduction-to-open-science-wednesday-18-may-1-pm-cest-20220518",title:"Webinar: Introduction to Open Science | Wednesday 18 May, 1 PM CEST"},{slug:"step-in-the-right-direction-intechopen-launches-a-portfolio-of-open-science-journals-20220414",title:"Step in the Right Direction: IntechOpen Launches a Portfolio of Open Science Journals"},{slug:"let-s-meet-at-london-book-fair-5-7-april-2022-olympia-london-20220321",title:"Let’s meet at London Book Fair, 5-7 April 2022, Olympia London"},{slug:"50-books-published-as-part-of-intechopen-and-knowledge-unlatched-ku-collaboration-20220316",title:"50 Books published as part of IntechOpen and Knowledge Unlatched (KU) Collaboration"},{slug:"intechopen-joins-the-united-nations-sustainable-development-goals-publishers-compact-20221702",title:"IntechOpen joins the United Nations Sustainable Development Goals Publishers Compact"},{slug:"intechopen-signs-exclusive-representation-agreement-with-lsr-libros-servicios-y-representaciones-s-a-de-c-v-20211123",title:"IntechOpen Signs Exclusive Representation Agreement with LSR Libros Servicios y Representaciones S.A. de C.V"},{slug:"intechopen-expands-partnership-with-research4life-20211110",title:"IntechOpen Expands Partnership with Research4Life"},{slug:"introducing-intechopen-book-series-a-new-publishing-format-for-oa-books-20210915",title:"Introducing IntechOpen Book Series - A New Publishing Format for OA Books"}]},book:{item:{type:"book",id:"10891",leadTitle:null,fullTitle:"Leishmaniasis - General Aspects of a Stigmatized Disease",title:"Leishmaniasis",subtitle:"General Aspects of a Stigmatized Disease",reviewType:"peer-reviewed",abstract:"Leishmaniasis is a major global health challenge, affecting approximately 12 million of the poorest people in 100 countries. It is a deforming and fatal disease in the visceral form. Therapies for leishmaniasis are numerically restricted, basically consisting of the administration of miltefosine, pentavalent antimonials, amphotericin B, or pentamidine. This is an important vulnerability against therapy efficiency that must be overcome by the scientific community. This book discusses important aspects of the disease, such as treatment, epidemiology, and molecular and cell biology. The information contained herein is important for young researchers as they seek to develop safe and effective treatments for this neglected tropical disease.",isbn:"978-1-83968-082-3",printIsbn:"978-1-83968-081-6",pdfIsbn:"978-1-83968-092-2",doi:"10.5772/intechopen.95200",price:119,priceEur:129,priceUsd:155,slug:"leishmaniasis-general-aspects-of-a-stigmatized-disease",numberOfPages:256,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"e2123440fdf88dfa14b4713a1a8ba58a",bookSignature:"Leonardo de Azevedo Calderonon",publishedDate:"April 13th 2022",coverURL:"https://cdn.intechopen.com/books/images_new/10891.jpg",numberOfDownloads:1198,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:0,numberOfDimensionsCitationsByBook:0,hasAltmetrics:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"May 5th 2021",dateEndSecondStepPublish:"June 2nd 2021",dateEndThirdStepPublish:"August 1st 2021",dateEndFourthStepPublish:"October 20th 2021",dateEndFifthStepPublish:"December 19th 2021",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"177382",title:"Dr.",name:"Leonardo de Azevedo",middleName:null,surname:"Calderon",slug:"leonardo-de-azevedo-calderon",fullName:"Leonardo de Azevedo Calderon",profilePictureURL:"https://mts.intechopen.com/storage/users/177382/images/system/177382.jpg",biography:"Leonardo de Azevedo Calderon obtained a bachelor\\'s degree and a doctorate in Biological Sciences from the University of Brasília (UnB), Brazil, in 2000 and 2004, respectively. He is a researcher in public health at the Oswaldo Cruz Foundation, lecturer in the Medicine Department, Federal University of Rondônia, Brazil, and head of the Center for the Study of Biomolecules Applied to Health. He has experience in the fields of biochemistry, molecular biophysics, and bioinformatics, working mainly in the prospection and development of new chemical entities with potentially useful activities in future therapies for leishmaniasis and malaria.",institutionString:"Oswaldo Cruz Foundation",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Oswaldo Cruz Foundation",institutionURL:null,country:{name:"Brazil"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"1046",title:"Infectious Diseases",slug:"infectious-diseases"}],chapters:[{id:"80467",title:"Leishmaniasis: Molecular Aspects of Parasite Dimorphic Forms Life Cycle",doi:"10.5772/intechopen.102370",slug:"leishmaniasis-molecular-aspects-of-parasite-dimorphic-forms-life-cycle",totalDownloads:77,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"According to WHO, Leishmaniasis is a complex neglected disease caused by a protozoa parasite from over 20 Leishmania species transmitted by more than 90 sandfly species, showing three main forms: visceral, cutaneous, and mucocutaneous. The efficient prevention and control of leishmaniasis are very difficult to achieve, depending on the combination of different intervention strategies, usually resulting in failure. Additionally, the correct diagnostics require the combination of clinical signs with laboratory tests, and only a few therapeutical options are available for patients. To improve this scenario, greater efforts in research for control and treatment are needed. For this purpose, the study and understanding of the life cycle of Leishmania are mandatory for all researchers who intend to dedicate their careers to the different aspects of this important disease. In order to support beginning researchers in the study of leishmaniasis, we propose in this review an update in the current knowledge about the major molecular aspects involved in the development of dimorphic forms of Leishmania parasites that replicate in the gut of sandflies (promastigotes) and in mammalian cells (amastigotes) and the relationship with host’s immune system.",signatures:"Natanael Endrew Souto Maior Torres Bonfim, Ana Lígia Barbour Scott and Leonardo de Azevedo Calderon",downloadPdfUrl:"/chapter/pdf-download/80467",previewPdfUrl:"/chapter/pdf-preview/80467",authors:[{id:"177382",title:"Dr.",name:"Leonardo de Azevedo",surname:"Calderon",slug:"leonardo-de-azevedo-calderon",fullName:"Leonardo de Azevedo Calderon"},{id:"465571",title:"Dr.",name:"Natanael",surname:"Endrew Souto Maior Torres Bonfim",slug:"natanael-endrew-souto-maior-torres-bonfim",fullName:"Natanael Endrew Souto Maior Torres Bonfim"},{id:"465572",title:"Dr.",name:"Ana",surname:"Lígia Barbour Scott",slug:"ana-ligia-barbour-scott",fullName:"Ana Lígia Barbour Scott"}],corrections:null},{id:"79372",title:"Protective and Pathogenic Immune Responses to Cutaneous Leishmaniasis",doi:"10.5772/intechopen.101160",slug:"protective-and-pathogenic-immune-responses-to-cutaneous-leishmaniasis",totalDownloads:85,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Leishmania (Kinetoplastida: Trypanosomatidae) parasites are known to cause a broad spectrum of clinical diseases in humans, collectively known as the leishmaniases. Cutaneous leishmaniasis is the most common clinical presentation with varying degrees of severity largely driven by host immune responses, specifically the interplay between innate and adaptive immune response. The establishment of a T lymphocyte driven cell-mediated immune response, leading to activated phagocytic cells, leading to Leishmania parasite killing and control of infection. Alternatively, the Leishmania parasite manipulates the host immune system, enabling parasite proliferation and clinical disease. Here we review how the cumulative interactions of different aspects of the host immune response determines disease outcome, severity, and immunity to re-infection.",signatures:"Elina Panahi, Danielle I. Stanisic, Christopher S. Peacock and Lara J. Herrero",downloadPdfUrl:"/chapter/pdf-download/79372",previewPdfUrl:"/chapter/pdf-preview/79372",authors:[{id:"80655",title:"Dr.",name:"Lara J.",surname:"Herrero",slug:"lara-j.-herrero",fullName:"Lara J. Herrero"},{id:"436057",title:"Ms.",name:"Elina",surname:"Panahi",slug:"elina-panahi",fullName:"Elina Panahi"},{id:"436058",title:"Dr.",name:"Christopher S.",surname:"Peacock",slug:"christopher-s.-peacock",fullName:"Christopher S. Peacock"},{id:"436059",title:"Dr.",name:"Danielle I.",surname:"Stanisic",slug:"danielle-i.-stanisic",fullName:"Danielle I. Stanisic"}],corrections:null},{id:"79300",title:"Extracellular Vesicles Released by Leishmania: Impact on Disease Development and Immune System Cells",doi:"10.5772/intechopen.101151",slug:"extracellular-vesicles-released-by-em-leishmania-em-impact-on-disease-development-and-immune-system-",totalDownloads:145,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Leishmania spp. release extracellular vesicles (EVs) containing parasite molecules, including several antigens and virulence factors. These EVs can interact with the host cells, such as immune cells, contributing to the parasite–host relationship. Studies have demonstrated that Leishmania-EVs can promote infection in experimental models and modulate the immune response. Although the immunomodulatory effect has been demonstrated, Leishmania-EVs can deliver parasite antigens and therefore have the potential for use as a new diagnostic tool and development of new therapeutic and vaccine approaches. This review aims to bring significant advances in the field of extracellular vesicles and Leishmania, focusing on their role in the cells of the immune system.",signatures:"Rogéria Cristina Zauli, Andrey Sladkevicius Vidal, Talita Vieira Dupin, Aline Correia Costa de Morais, Wagner Luiz Batista and Patricia Xander",downloadPdfUrl:"/chapter/pdf-download/79300",previewPdfUrl:"/chapter/pdf-preview/79300",authors:[{id:"421426",title:"Prof.",name:"Patricia",surname:"Xander",slug:"patricia-xander",fullName:"Patricia Xander"},{id:"421431",title:"Ms.",name:"Andrey",surname:"Sladkevicius Vidal",slug:"andrey-sladkevicius-vidal",fullName:"Andrey Sladkevicius Vidal"},{id:"421432",title:"Ms.",name:"Talita",surname:"Vieira Dupin",slug:"talita-vieira-dupin",fullName:"Talita Vieira Dupin"},{id:"421433",title:"Ms.",name:"Aline",surname:"Correia Costa de Morais",slug:"aline-correia-costa-de-morais",fullName:"Aline Correia Costa de Morais"},{id:"421434",title:"MSc.",name:"Rogéria Cristina",surname:"Zauli",slug:"rogeria-cristina-zauli",fullName:"Rogéria Cristina Zauli"},{id:"421435",title:"Prof.",name:"Wagner",surname:"Luiz Batista",slug:"wagner-luiz-batista",fullName:"Wagner Luiz Batista"}],corrections:null},{id:"79348",title:"Treatment of Leishmaniasis",doi:"10.5772/intechopen.101145",slug:"treatment-of-leishmaniasis",totalDownloads:58,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Treatment of Leishmaniasis is always not satisfactory despite advancement all these years. This chapter will discuss the standard treatment options like Amphotericin, oral miltefosine, topical paromycin, add more details about newer emerging drugs and alternative therapies, surgical treatment modalities for resistant cases. Will discuss few information regarding vaccines. Special precautions to be taken while travelling to endemic areas will be discussed. Management protocol for mucocutaneous and visceral type will be highlighted. Side-effects of drugs used in the treatment of Leishmaniasis will be discussed in short and measures to monitor these side effects will be discussed. Long term monitoring of relapse will also be discussed.",signatures:"R. Sivayogana, Aishwarya Krishnakumar, S. Kumaravel, Rajesh Rajagopal and P. Ravikanth",downloadPdfUrl:"/chapter/pdf-download/79348",previewPdfUrl:"/chapter/pdf-preview/79348",authors:[{id:"419522",title:"Assistant Prof.",name:"R.",surname:"Sivayogana",slug:"r.-sivayogana",fullName:"R. Sivayogana"},{id:"429065",title:"Dr.",name:"Aishwarya",surname:"Krishnakumar",slug:"aishwarya-krishnakumar",fullName:"Aishwarya Krishnakumar"},{id:"440470",title:"Dr.",name:"S.",surname:"Kumaravel",slug:"s.-kumaravel",fullName:"S. Kumaravel"},{id:"455626",title:"Associate Prof.",name:"Rajesh",surname:"Rajagopal",slug:"rajesh-rajagopal",fullName:"Rajesh Rajagopal"},{id:"455627",title:"Dr.",name:"P.",surname:"Ravikanth",slug:"p.-ravikanth",fullName:"P. Ravikanth"}],corrections:null},{id:"79223",title:"Visceral Leishmaniasis: Asymptomatic Facts",doi:"10.5772/intechopen.101109",slug:"visceral-leishmaniasis-asymptomatic-facts",totalDownloads:128,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Visceral Leishmaniasis (VL) caused by protozoan parasite Leishmania is a vector borne disease and infection is limited not to human but also to animals worldwide. For infection identification and prevalence in both Leishmania endemic and nonendemic regions, several serological and genetic techniques are used. Although diagnostic techniques and clinical symptoms can establish illness status, it is extremely difficult to diagnose infection in the absence of symptoms. Asymptomatic are healthy people who have an infection but are unaware of it. The epidemiology of asymptomatic Leishmaniasis is critical for its eradication. Only a small percentage of infected people are clinically suspected of having VL, as the majority of them may not show any symptoms and remain asymptomatic. Some asymptomatic infections may go away after a while, or they may linger for years, or they may develop to illness with clinical signs. Asymptomatic infection varies per endemic location, but almost all of them point to this hidden category of parasite infection. It is now critical to understand many factors such as diagnostic markers, genetic markers, and immunological markers along with different risk factors. All of these criteria, as well as some innovative techniques to diagnosing and controlling asymptomatic leishmaniasis, will be covered in this chapter. The main focus will be on asymptomatic condition of Indian Visceral Leishmaniasis, which is caused by Leishmania donovani and spreads via female sand fly P. argentipes biting. The numerous criteria that play a role in asymptomatic to symptomatic conversion in a specific time period will also be discussed in this chapter.",signatures:"Medhavi Sudarshan and Sumit Sharan",downloadPdfUrl:"/chapter/pdf-download/79223",previewPdfUrl:"/chapter/pdf-preview/79223",authors:[{id:"335269",title:"Assistant Prof.",name:"Medhavi",surname:"Sudarshan",slug:"medhavi-sudarshan",fullName:"Medhavi Sudarshan"},{id:"440228",title:"Dr.",name:"Sumit",surname:"Sharan",slug:"sumit-sharan",fullName:"Sumit Sharan"}],corrections:null},{id:"80815",title:"Geographical Distribution of Cutaneous Leishmaniasis and Pathogenesis",doi:"10.5772/intechopen.101841",slug:"geographical-distribution-of-cutaneous-leishmaniasis-and-pathogenesis",totalDownloads:68,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Leishmaniasis is still considered to be a global health problem, which spreads in most countries in the world. Leishmania is an intracellular obligate protistan parasite that causes different clinical symptoms in infected humans and other animals. There are clinically different types of the disease including: visceral, cutaneous or muco-cutaneous leishmaniasis. Approximately, two million new infections occurring annually; 0.7 to 1.2 million cases are recorded with cutaneous leishmaniasis and 200,000–400,000 cases return for visceral leishmaniasis. However, Cutaneous leishmaniasis considers one of uncontrolled wobbling endemic diseases, especially in Iraq, which occurs at the skin to cause a dermal lesion. Usually, the lesion is spontaneously healed to leave a colorless depressed scar and permanent immunity.",signatures:"Mohammed Hassan Flaih",downloadPdfUrl:"/chapter/pdf-download/80815",previewPdfUrl:"/chapter/pdf-preview/80815",authors:[{id:"419546",title:"Dr.",name:"Mohammed",surname:"Hassan Flaih",slug:"mohammed-hassan-flaih",fullName:"Mohammed Hassan Flaih"}],corrections:null},{id:"79230",title:"Geopolitical and Geospatial Conflicts Affecting Cutaneous Leishmaniasis: Iraqi Cases, 2014-2015",doi:"10.5772/intechopen.101147",slug:"geopolitical-and-geospatial-conflicts-affecting-cutaneous-leishmaniasis-iraqi-cases-2014-2015",totalDownloads:47,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"In Iraq, increment of cutaneous leishmaniasis (CL) was reported, a zoonotic type caused by Leishmania major and an anthroponotic type caused by Leishmania tropica. This is attributed to environmental changes, irregular construction, increased waste, and stray dogs in the cities. Internal displacement, poverty, and malnutrition play a role in the occurrence of disease. Iraq has been exposed to wars, civil war, widespread violence, and terrorism, Internal displacement of population, unlawful housing, shortage of municipal services, and accumulation of garbage have been increasing in the cities since 2003. Reports from Iraqi MOH documented an increase in the rate of CL. Eventually, case investigation, vector, and reservoir controls were suspended. Geopolitical conflicts and geospatial health deterioration contributed to an increase in various dominant reservoir species in these areas, particularly for ZCL, and the activity of the sand flies. A sudden sharp increase in CL cases was observed during 2003–2015. In conclusion, ongoing crises exposed Iraq to deterioration, collapse, and destruction of health system infrastructure and uncontrolled urbanization, all of which can act as risk factors for Leishmaniasis.",signatures:"Karim Abdulkadim Muftin Al Zadawi",downloadPdfUrl:"/chapter/pdf-download/79230",previewPdfUrl:"/chapter/pdf-preview/79230",authors:[{id:"419629",title:"Dr.",name:"Karim",surname:"Abdulkadim Muftin Al Zadawi",slug:"karim-abdulkadim-muftin-al-zadawi",fullName:"Karim Abdulkadim Muftin Al Zadawi"}],corrections:null},{id:"79857",title:"Use of Cell Biology to Identify Cellular Targets in Drug Development Process against Leishmania Sp.",doi:"10.5772/intechopen.101662",slug:"use-of-cell-biology-to-identify-cellular-targets-in-drug-development-process-against-em-leishmania-e",totalDownloads:136,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Leishmaniasis is one of the most important neglected tropical diseases. The chemotherapy for its treatment uses very toxic compounds with a low efficacy rate. Thus, there is an urgent need to develop new chemotherapeutic agents to help countries control this devasting disease. In drug development, different approaches can be used to identify potential cellular targets that allow us to understand better the cell biology of eukaryotic cells. Several groups are dedicated to studying new molecules, searching for promising candidates against Leishmania. Different techniques have been used to characterize the cell biology, biochemistry, and molecular biology alterations induced by the treatments, trying to understand the mechanisms of action. The main goal of this chapter is to describe an overview of the literature exploring the several studies published about the chemotherapy of anti-Leishmania concerning the mechanisms of action of different classes of molecules or therapeutic alternatives.",signatures:"Gabrielle dos Santos da Silva e Miranda, Joseane Lima Prado Godinho, Sara Teixeira de Macedo-Silva, Brunno Renato Farias Verçoza, Alisson Amaral da Rocha, Milena Barenco Pires de Abreu Sodré, Victor Feliciano dos Santos Ramos and Juliany Cola Fernandes Rodrigues",downloadPdfUrl:"/chapter/pdf-download/79857",previewPdfUrl:"/chapter/pdf-preview/79857",authors:[{id:"420224",title:"Associate Prof.",name:"Juliany",surname:"Cola Fernandes Rodrigues",slug:"juliany-cola-fernandes-rodrigues",fullName:"Juliany Cola Fernandes Rodrigues"},{id:"420943",title:"Ms.",name:"Gabrielle",surname:"dos Santos da Silva e Miranda",slug:"gabrielle-dos-santos-da-silva-e-miranda",fullName:"Gabrielle dos Santos da Silva e Miranda"},{id:"420944",title:"Dr.",name:"Brunno Renato",surname:"Farias Verçoza",slug:"brunno-renato-farias-vercoza",fullName:"Brunno Renato Farias Verçoza"},{id:"420945",title:"Mr.",name:"Alisson",surname:"Amaral Da Rocha",slug:"alisson-amaral-da-rocha",fullName:"Alisson Amaral Da Rocha"},{id:"421145",title:"Dr.",name:"Joseane",surname:"Lima Prado Godinho",slug:"joseane-lima-prado-godinho",fullName:"Joseane Lima Prado Godinho"},{id:"421148",title:"Dr.",name:"Sara",surname:"Teixeira de Macedo-Silva",slug:"sara-teixeira-de-macedo-silva",fullName:"Sara Teixeira de Macedo-Silva"},{id:"421152",title:"Ms.",name:"Milena",surname:"Barenco Pires De Abreu Sodré",slug:"milena-barenco-pires-de-abreu-sodre",fullName:"Milena Barenco Pires De Abreu Sodré"},{id:"421153",title:"Mr.",name:"Victor",surname:"Feliciano Dos Santos Ramos",slug:"victor-feliciano-dos-santos-ramos",fullName:"Victor Feliciano Dos Santos Ramos"}],corrections:null},{id:"79559",title:"Toward New Antileishmanial Compounds: Molecular Targets for Leishmaniasis Treatment",doi:"10.5772/intechopen.101132",slug:"toward-new-antileishmanial-compounds-molecular-targets-for-leishmaniasis-treatment",totalDownloads:119,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The leishmaniases are a group of diseases caused by protozoan parasites—Leishmania sp. Leishmaniasis is classified among the 20 neglected diseases by WHO. Although the disease has been known for more than 120 years, the number of drugs used for the treatment is still limited to 5–6. The first-line drugs against leishmaniasis are pentavalent antimonials, which were introduced to the treatment 70 years ago—despite all their side effects. Molecular targets are becoming increasingly important for efficacy and selectivity in postgenomic drug research studies. In this chapter, we have discussed potential therapeutic targets of antileishmanial drug discovery such as pteridine reductase (PTR1), trypanothione reductase (TR), N-myristoyltransferase (NMT), trypanothione synthetase (TryS), IU-nucleoside hydrolase, and topoisomerases, enzymes and their inhibitors reported in the literature.",signatures:"Huseyin Istanbullu and Gulsah Bayraktar",downloadPdfUrl:"/chapter/pdf-download/79559",previewPdfUrl:"/chapter/pdf-preview/79559",authors:[{id:"419167",title:"Assistant Prof.",name:"Huseyin",surname:"Istanbullu",slug:"huseyin-istanbullu",fullName:"Huseyin Istanbullu"},{id:"435370",title:"Dr.",name:"Gulsah",surname:"Bayraktar",slug:"gulsah-bayraktar",fullName:"Gulsah Bayraktar"}],corrections:null},{id:"79649",title:"microRNAs: Are They Important in the Development of Resistance in Leishmaniasis?",doi:"10.5772/intechopen.101514",slug:"micrornas-are-they-important-in-the-development-of-resistance-in-leishmaniasis-",totalDownloads:176,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Leishmaniasis is an infectious and parasitic disease of great importance in public health. Numerous studies indicate that biochemical and molecular mechanisms are factors that contribute to the emergence of antileishmanial drug resistance. Currently, miRNAs have been identified as targets for the invasion of pathogens to control the immune response and imply resistance to treatments. Considering the alarming growth in drug resistance, new possibilities for controlling leishmaniasis have been emerging. Natural compounds originating from medicinal plants are being increasingly explored as promising antileishmanial alternatives. The chapter aims to provide a brief review on mechanisms of action associated with traditional agents used to treat leishmaniasis, focusing mainly on molecular bases associated with the resistance of Leishmania spp. to current drugs and identifying the possible miRNAs involved in this process. In addition, we seek to describe some of the promising plant molecules that can be used as potential antileishmanial agents and their possible mechanisms of action.",signatures:"Sandra Alves de Araújo, Tatiane Aranha da Penha-Silva, Jaqueline Diniz Pinho, Marcelo de Souza Andrade and Ana Lucia Abreu-Silva",downloadPdfUrl:"/chapter/pdf-download/79649",previewPdfUrl:"/chapter/pdf-preview/79649",authors:[{id:"223173",title:"Dr.",name:"Ana Lucia",surname:"Abreu-Silva",slug:"ana-lucia-abreu-silva",fullName:"Ana Lucia Abreu-Silva"},{id:"233684",title:"Dr.",name:"Tatiane",surname:"Aranha da Penha-Silva",slug:"tatiane-aranha-da-penha-silva",fullName:"Tatiane Aranha da Penha-Silva"},{id:"421473",title:"MSc.",name:"Sandra",surname:"Alves de Araújo",slug:"sandra-alves-de-araujo",fullName:"Sandra Alves de Araújo"},{id:"428524",title:"Dr.",name:"Jaqueline",surname:"Diniz Pinho",slug:"jaqueline-diniz-pinho",fullName:"Jaqueline Diniz Pinho"},{id:"428526",title:"Dr.",name:"Marcelo",surname:"de Souza Andrade",slug:"marcelo-de-souza-andrade",fullName:"Marcelo de Souza Andrade"}],corrections:null},{id:"79718",title:"Molecular Informatics of Trypanothione Reductase of Leishmania major Reveals Novel Chromen-2-One Analogues as Potential Leishmanicides",doi:"10.5772/intechopen.100594",slug:"molecular-informatics-of-trypanothione-reductase-of-em-leishmania-major-em-reveals-novel-chromen-2-o",totalDownloads:135,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Trypanothione reductase (TR), a flavoprotein oxidoreductase is an important therapeutic target for leishmaniasis. Ligand-based pharmacophore modelling and molecular docking were used to predict selective inhibitors against TR. Homology modelling was employed to generate a three-dimensional structure of Leishmania major trypanothione reductase (LmTR). A pharmacophore model used to screen a natural compound library generated 42 hits, which were docked against the LmTR protein. Compounds with lower binding energies were evaluated via in silico pharmacological profiling and bioactivity. Four compounds emerged as potential leads comprising Karatavicinol (7-[(2E,6E,10S)-10,11-dihydroxy-3,7,11-trimethyldodeca-2,6-dienoxy]chromen-2-one), Marmin (7-[(E,6R)-6,7-dihydroxy-3,7-dimethyloct-2-enoxy]chromen-2-one), Colladonin (7-[[(4aS)-6-hydroxy-5,5,8a-trimethyl-2-methylidene-3,4,4a,6,7,8-hexahydro-1H-naphthalen-1-yl]methoxy]chromen-2-one), and Pectachol (7-[(6-hydroxy-5,5,8a-trimethyl-2-methylidene-3,4,4a,6,7,8-hexahydro-1H-naphthalen-1-yl)methoxy]-6,8-dimethoxychromen-2-one) with good binding energies of −9.4, −9.3, 8.8, and −8.5 kcal/mol, respectively. These compounds bound effectively to the FAD domain of the protein with some critical residues including Asp35, Thr51, Lys61, Tyr198, and Asp327. Furthermore, molecular dynamics simulations and molecular mechanics Poisson-Boltzmann surface area (MMPBSA) computations corroborated their strong binding. The compounds were also predicted to possess anti-leishmanial activity. The molecules serves as templates for the design of potential drug candidates and can be evaluated in vitro with optimistic results in producing plausible attenuating infectivity in macrophages.",signatures:"Samuel K. Kwofie, Gabriel B. Kwarko, Emmanuel Broni, Michael B. Adinortey and Michael D. Wilson",downloadPdfUrl:"/chapter/pdf-download/79718",previewPdfUrl:"/chapter/pdf-preview/79718",authors:[{id:"419961",title:"Dr.",name:"Samuel K.",surname:"Kwofie",slug:"samuel-k.-kwofie",fullName:"Samuel K. Kwofie"},{id:"429019",title:"Mr.",name:"Gabriel B.",surname:"Kwarko",slug:"gabriel-b.-kwarko",fullName:"Gabriel B. Kwarko"},{id:"429020",title:"Dr.",name:"Emmanuel",surname:"Broni",slug:"emmanuel-broni",fullName:"Emmanuel Broni"},{id:"429021",title:"Prof.",name:"Michael B.",surname:"Adinortey",slug:"michael-b.-adinortey",fullName:"Michael B. Adinortey"},{id:"429022",title:"Prof.",name:"Michael D.",surname:"Wilson",slug:"michael-d.-wilson",fullName:"Michael D. Wilson"}],corrections:null},{id:"80026",title:"Plant-Based Alternative Treatment for Leishmaniasis: A Neglected Tropical Disease",doi:"10.5772/intechopen.101958",slug:"plant-based-alternative-treatment-for-leishmaniasis-a-neglected-tropical-disease",totalDownloads:29,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Leishmaniasis is a third most important vector born disease caused by intracellular parasite belongs to genus Leishmania. The leishmaniasis is prevalent in 102 countries/areas worldwide. Approximately, it effected 350 million people worldwide. Leishmaniasis effects developing and undeveloped countries globally. Antileishmanial drugs (pentavalent antimonials, stibogluconate, miltefosine, paramycin, and amphotericin) are most vital tool for curing leishmaniasis. However, none of these drugs is free from side effect including cost, toxicity, drug resistance, administration route, and prolong time, these disadvantages are main obstacle in the Leishmania infection eradication. Considering the increasing cases of leishmaniasis and drug resistance there is an urgent need for an effective and novel approach against leishmaniasis. Therefore, many researchers have tried to develop new medicines for the treatment of Leishmania infection. In the course of new therapies identification, plant based compounds were found to be an alternative that can be either used directly or with structural modifications. Several plants have been known for ages to be the source of phytochemicals with high values of medicines. These phytochemicals have been extracted by various techniques and have shown efficacy for the curing of several diseases. This chapter study explain various applications based on green approaches drugs for the treatment of leishmaniasis.",signatures:"Nargis Shaheen, Chaitenya Verma and Naveeda Akhter Qureshi",downloadPdfUrl:"/chapter/pdf-download/80026",previewPdfUrl:"/chapter/pdf-preview/80026",authors:[{id:"224439",title:"Dr.",name:"Naveeda",surname:"Akhter Qureshi",slug:"naveeda-akhter-qureshi",fullName:"Naveeda Akhter Qureshi"},{id:"420059",title:"Dr.",name:"Nargis",surname:"Shaheen",slug:"nargis-shaheen",fullName:"Nargis Shaheen"},{id:"420980",title:"Dr.",name:"Chaitenya",surname:"Verma",slug:"chaitenya-verma",fullName:"Chaitenya Verma"}],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},subseries:null,tags:null},relatedBooks:[{type:"book",id:"2327",title:"Chromatography",subtitle:"The Most Versatile Method of Chemical Analysis",isOpenForSubmission:!1,hash:"730549acc4ff27929bb106ba63661172",slug:"chromatography-the-most-versatile-method-of-chemical-analysis",bookSignature:"Leonardo de Azevedo Calderon",coverURL:"https://cdn.intechopen.com/books/images_new/2327.jpg",editedByType:"Edited by",editors:[{id:"177382",title:"Dr.",name:"Leonardo de Azevedo",surname:"Calderon",slug:"leonardo-de-azevedo-calderon",fullName:"Leonardo de Azevedo Calderon"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,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"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"825",title:"Current Topics in Tropical Medicine",subtitle:null,isOpenForSubmission:!1,hash:"ef65e8eb7a2ada65f2bc939aa73009e3",slug:"current-topics-in-tropical-medicine",bookSignature:"Alfonso J. 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\r\n\tSatellite radar altimetry, with its global coverage and level of precision, has been a key technique for observing the ocean surface and many aspects of land surfaces over the last three decades. Together with Earth sciences, it has integrated the efforts of international researchers to demonstrate the techniques, missions, measurements, and satellite altimetry accuracy, including orbit determination and ocean circulation models. Satellite altimetry has revolutionized our understanding of the Earth’s sea-level shape and its change over time, monitoring the natural and human-induced water mass balance change, marine gravity computations, and high-resolution seafloor bathymetry reconstruction, plate tectonics, etc., thus providing significant impact for climate and environmental studies. For researchers in geodesy and geophysics, oceanography, and other space and earth sciences, satellite radar altimetry is critical for unifying the vertical height systems, regional and global geoid modelling, and monitoring the sea level rise impact but also the ice sheet melting and others. This book gives a state-of-the-art overview of the technology itself and recent developments, including the applied studies utilizing the altimeter data for ice sheet monitoring, vertical land motion estimating, bathymetric computations, and marine geoid modelling, but also Synthetic Aperture Radar altimetry, coastal altimetry tracking methods and new satellite missions (e.g., Sentinel-6).
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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. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"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:"314",title:"Regenerative Medicine and Tissue Engineering",subtitle:"Cells and Biomaterials",isOpenForSubmission:!1,hash:"bb67e80e480c86bb8315458012d65686",slug:"regenerative-medicine-and-tissue-engineering-cells-and-biomaterials",bookSignature:"Daniel Eberli",coverURL:"https://cdn.intechopen.com/books/images_new/314.jpg",editedByType:"Edited by",editors:[{id:"6495",title:"Dr.",name:"Daniel",surname:"Eberli",slug:"daniel-eberli",fullName:"Daniel Eberli"}],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:"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:"2270",title:"Fourier Transform",subtitle:"Materials Analysis",isOpenForSubmission:!1,hash:"5e094b066da527193e878e160b4772af",slug:"fourier-transform-materials-analysis",bookSignature:"Salih Mohammed Salih",coverURL:"https://cdn.intechopen.com/books/images_new/2270.jpg",editedByType:"Edited by",editors:[{id:"111691",title:"Dr.Ing.",name:"Salih",surname:"Salih",slug:"salih-salih",fullName:"Salih Salih"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"72535",title:"RNAi Induced Gene Silencing Journey from Simple dsRNA to High-Throughput Intron Hairpin RNA Construct in Crop Improvement",doi:"10.5772/intechopen.93012",slug:"rnai-induced-gene-silencing-journey-from-simple-dsrna-to-high-throughput-intron-hairpin-rna-construc",body:'The introduction of recombinant DNA techniques and the progression in gene transfer technique and plant regeneration strategies in the 1980s empowered the trading of genes not only between plants of various species but also from viruses, bacteria, fungi, and animals into plants. This capacity to transform plants with transgenes gave unparalleled chances for crop improvement and also widened the scope for upgrading of strategies in genetic engineering. One such strategy developed was RNAi for gene silencing.
The RNAi pathway was first observed in the nematode in response to double-stranded RNA (dsRNA) which resulted in sequence-specific gene silencing. It was then developed as weapon to fight an ongoing war between viruses and living organism from decades. Researchers have been constantly working toward combating virus invasion especially in crops. In this context in 1986, the American plant virologist Roger Beachy and his partners demonstrated an experiment against the tobacco mosaic virus (TMV) infection in tobacco plants. Using gene transformation technique, they introduced a transgene derived from the coat protein gene of tobacco mosaic infection in tobacco plants. This virus-based transgene acted against the TMV and protected plant from infection [1]. This finding inspired significant research movement around the globe and prompted numerous reports of protection against a range of viruses infecting plants. However, the underlying mechanism of resistance against viruses using pathogen-derived gene at the molecular level was not known at that time.
The study of Lindbo and Dougherty [2] gave an insight to the happenings at the molecular level. They show that transforming plants with transgenes that can express RNAs of viral origin, but which could not be translated into proteins, could initiate a mechanism that could degrade both the mRNA from the introduced transgene and the genomic RNA of the inoculated virus and lead to repression of the infection by the virus. They also found great similarities with a phenomenon called posttranscriptional gene silencing (PTGS)—which had been described using transgenes of nonviral origin.
Transgene-induced posttranscriptional gene silencing has been detected in plants [3] where it is called co-suppression and in fungi where it is called quelling [4]. In animals, PTGS can be induced by dsRNA in a process called RNA interference (RNAi) [5]. In plants, PTGS can also be induced by viruses expressing host genes in a process called virus-induced gene silencing (VIGS) [6]. Viruses themselves can be the targets of the PTGS machinery [6]. It was proposed that if a cell expresses molecules of the same RNA sequence at a level higher than a certain threshold, this induces a mechanism that specifically destroys that RNA [7]. This proposition led to changes in the way of construct designing and preparation, and the journey from dsRNA construct to high-throughput ihpRNA construct began.
Based on the Lindbo and Dougherty proposition [2], constructs were made with an aim to increase the copy number of virus-derived transgenes [8]. Constructs contained two sense copies or two antisense copies or contained both one sense and one antisense copy. The transgenic plants with these different constructs were analyzed for virus resistance. They observed that tobacco plants transformed with construct containing both sense and antisense copies showed higher protection against PVY virus. Based on the analysis, they concluded that the production of dsRNA was causing the silencing effect and not the number of gene copies.
Hence, the scientist thought of introducing dsRNA of the target RNA in plant to trigger the silencing mechanism. An experiment was conducted to induce immunity against potato virus Y (PVY) in tobacco plant [8]. Tobacco plants were transformed with gene constructs that encoded the Pro sequence in the sense or antisense orientation or in both orientations and were challenged with PVY. Less than 15% of the Pro[s] or Pro[a] lines showed resistance to PVY, whereas the lines containing both sense and antisense genes showed 44–54% of resistance to PVY. This result suggests that the sense and antisense mRNA in the same cell elicit the PVY resistance. Similar reports were obtained in rice cells transformed with a GUS-derived hairpin (hp) gene. The cells with the GUS gene alone gave a strong blue color, but those with both the GUS gene and the GUS-hp gene remained white [9].
Further the construct was improved with a transgene expressing RNA that folds back and hybridizes with itself to form a structure like a hairpin, instead of making two separate RNAs which hybridize into dsRNA [8, 9, 10, 11]. With ongoing research and use of RNAi technique for gene silencing, it was observed that silencing efficiency of RNAi vectors can be increased if the factors like selection of target sequences, the inverted repeats, size of the repeats, vectors, use of spacers or introns in the RNAi cassette, promoters, etc. are efficiently worked upon in vector construction. Studies on these parameters were conducted for improvement of hpRNA construct to increase silencing efficiency.
Overall efficiency of RNAi is dependent on RNA-protein interactions during siRNARISC assembly and activation [12, 13]. Hence, a systematic analysis of 180 siRNAs targeting the mRNA of two genes was done to identify siRNA-specific features that are likely to contribute to efficient processing at each step of RNAi [14]. Based on the analysis, characters like low G/C content, a bias toward low internal stability at the sense strand 3′-terminus, lack of inverted repeats, sense strand base preferences (positions 3, 10, 13, and 19), etc. were found to be affecting siRNA functionality. An algorithm was designed to incorporate all the eight criteria for the selection of potent siRNA for facilitating the functional gene knockdown studies.
A similar study was conducted to decipher the relationship between short interfering RNA (siRNA) sequence and RNA interference in three mammalian and
The investigation was done on the use of RNA interference for obtaining resistance against Cotton leaf curl Multan virus (CLCuMV) [16]. Three hairpin RNAi constructs were produced containing either complementary-sense genes essential for replication/pathogenicity or noncoding regulatory sequences of CLCuMV. All three RNAi constructs significantly reduced the replication of the virus in inoculated tissues. However, the systemic movement of the virus was controlled by only one of the constructs (CLCRNAiRepTrAPREn/pFGC), possibly because it spanned three virus-encoded genes: the replication-associated protein (Rep), the replication enhancer protein (Ren), and the transcriptional activator protein (TrAP). Also, the ability of virus to infect plant was compromised as the expression of TrAP was downregulated. TrAP is a transcription factor possibly involved in suppression of silencing machinery. Hence, both the target sequence and the levels of identity between the construct and target sequence determined the outcome of RNAi-based resistance against the virus.
On studying the effects of the structure, position, and sequence of a target RNA on RNAi using 47 constructs for inhibition of firefly luciferase activities by siRNAs targeted to TAR motif, it was observed that the efficacy of siRNAs depended mainly on the target sequence [17]. Statistical analysis of the data collected on the sequence preferences indicated that some nucleotides at specific positions are positively or negatively correlated with the efficiencies of siRNAs; for example, the siRNAs with an A residue at the 19th nucleotide position from the 5′ end of the sense strand showed relatively high suppressive activities, and siRNAs with a G residue at the 19th nucleotide in the sense strand tended to be less effective. Similar preference (A19 in siRNA; U1 in the miRNA) was observed for miRNA sequences [18]. This preference suggests the importance of the low internal stability of the 5′ terminus of the antisense strand and a possible functional contribution of a U at the 5′ end to the activities of both siRNA and miRNA [12, 13]. A significant negative correlation was also observed between the GC content of the 3′ half of siRNAs (in particular, from the 12th to the 19th nucleotide) and the activities of siRNAs. These tendencies can also be seen in another report [19].
The first commercialized transgenic papaya carrying the PRSV CP gene was introduced to Hawaii in 1998 to save the remains of the papaya industry [20]. However, transgenic papaya cultivars showed varying levels of resistance against PRSV isolates from other geographical regions. For example, isolates from the Bahamas, Florida, and Mexico have delayed, mild symptoms. Isolates from Brazil and Thailand also have delayed symptoms, but the virus eventually overcomes their resistance. The CP hemizygous line, “Rainbow,” is also susceptible to PRSV isolates from Taiwan [21]. Resistance levels therefore were found to be dependent on the variability among CP genes of the isolates [22, 23, 24]. The high levels of genetic divergence in PRSV isolates from Hainan caused the failure of transgenic papaya lines that targets specific viral CP gene [25]. CP-transgenic resistance of papaya is expressed in a nucleotide-sequence-homology-dependent manner [26, 27].
The proper selection of a target sequence for a given gene of interest remains one of the most critical components of successful gene knockdown regardless of the RNAi methodology.
The silencing efficiency is reported to be proportional to the size of target sequence, and the terminal regions (5′ and 3′) were found to be unaffected by silencing mechanism [28]. This description was based on the experiment conducted to identify the target regions and relative efficiencies of various target regions for silencing of gn1 (β-1,3 glucanase) gene in transgenic tobacco line T17.
Similarly, the effect of length of inverted repeats in tobacco BY-2 cells was tested by co-transformation of a luciferase gene construct and a luciferase dsRNA expression plasmid [29]. The dsRNA expression plasmids targeted to the firefly luciferase gene were constructed with 500- and 300-bp inverted repeats. However, no significant difference in silencing efficiency was observed, and the presence of 300-bp dsRNA was found to be sufficient to suppress the luciferase activity in cultured plant cells. A longer dsRNA did not show any enhancement in RNAi effect.
The less silencing frequency is reported when shorter fragments were used [30]. It was based on constructs with fragments of range 50 bp–1 kb targeted to silence two
Differences in silencing efficiency were also observed in
The effect of size of granule-bound starch synthase (GBSSI) sequence in inverted repeat constructs was evaluated, and it was found that, for GBSSI, the small inverted repeat constructs were more efficient silencing inducers than the large inverted repeat constructs. The small inverted repeat constructs with a repeat size of 500–600 bp and a spacer of about 150 bp were observed to be more efficient silencing inducers than the large inverted repeat constructs where the size of the repeat was 1.1 or 1.3 kb whilst the size of spacer was 1.3 or 1.1 kb [32].
Together, these reports suggest that 300–500 bp is the optimal size for inverted repeat for effective silencing mechanism.
Construction of RNAi vectors takes considerable time and is a tedious task since it involves laborious conventional cloning technology that relies on restriction digestion and ligation of two fragments corresponding to the antisense and sense region of the stem and subcloning into a binary vector. Hence, there is demand for high-throughput plant RNAi vectors for a rapid and easy construction.
For instance, technique involving a single step for construction of an RNAi vector has been developed that facilitates fast and reliable DNA cloning. It is called as gateway cloning technique which is also available with compatibility for
For the construction of RNAi vectors using Gateway recombination technology, the PCR products of the target gene are generated with primers flanking attB1 and attB2 sites for recombination with two cloning sites with attP1 and attP2 sequences using BP clonase. pHELLSGATE [30] and pANDA [30] are the vectors that allow the assembly of an inverted repeat structure by Gateway recombination technology [33]. Several pHELLSGATE-related RNAi vectors have been developed for RNAi in monocotyledonous plants and for inducible RNAi [34, 35]. For conventional cloning, pHANNIBAL [36], pKANNIBAL [37], pSAT [38], and pSH [39] are available. In these RNAi vectors, PCR fragments of the target gene are produced by using primers with restriction sites and cloned successively into both upstream and downstream regions of the spacer to become the two arms of the hairpin construct. Simultaneously work was done to enhance the efficiency in cloning inverted repeats for RNAi. An all-purpose vector, pGEMWIZ, for assembling the repeat for any
For a simple and efficient construction of intron-containing hpRNA (ihpRNA) vectors, a novel restriction-ligation approach was developed. The system was designed based on the type IIs restriction enzyme BsaI and plant RNAi vector pRNAi-GG based on the Golden Gate (GG) cloning [41]. It required only a single PCR product of the gene of interest flanked with BsaI recognition sequence which can then be cloned into pRNAi-GG at both sense and antisense orientations simultaneously to form ihpRNA construct. The ihpRNA construction process could be completed in one tube with one restriction-ligation step.
The silencing efficiency of RNAi vectors was reportedly enhanced by the addition of an intron interposed between the inverted flanking target sequences in vector. The spacer fragment was observed to give the stability to the inverted repeat sequences. It did not play any role in PTGS and was spliced out during pre-mRNA processing. The process of intron excision from the construct by the spliceosome might help to align the complementary arms of the hairpin in an environment favoring RNA hybridization and promoting the formation of a duplex. Also, splicing may contribute to increase the amount of hairpin RNA by preventing the hairpin’s passage from the nucleus or by creating a smaller, less nuclease-sensitive loop [10].
Smith et al. [10] showed that transgene constructs encoding a splicable intron within a hairpin RNA structure can induce PTGS with almost 100% efficiency. The percentage of PVY-resistant tobacco plants obtained by targeting the nuclear inclusion a (NIa) protease gene of PVY with different constructs was 7% for the sense gene, 4% for the antisense gene, 58% for the hpRNA with a nonspliceable loop separating the sense and antisense arms, and 96% for the same hpRNA with a spliceable intron. The high efficiency of intron hpRNA constructs for inducing PTGS and generating virus-immune transgenic plants was confirmed for Cucumber mosaic virus (CMV) [42, 43].
The effect of spacer sequences on silencing potential of RNAi constructs was demonstrated [44]. It was tested by an in vivo assay of the α-linolenic acid content in hairy roots of tobacco with RNAi vectors against ω-3 fatty acid desaturase (NtFAD3) gene responsible for production of α-linolenic acid of root membrane lipids. The frequency of RNA silencing was observed to be more affected by spacer sequences than by spacer length (100–1800 bp). They concluded that it is possible to change the degree of silencing by replacing spacer sequences. They predicted the reason to be interaction of spacer sequences with stem sequences of the hpRNA which affect the formation of a hairpin structure.
The degree of silencing is apparently proportional to the level of siRNAs, and the following “strong” promoters have been used in the construction of RNAi vectors: the cauliflower mosaic virus 35S promoter (p35S) [45, 46, 47], soybean lectin promoter [48],
RNAi technique cannot be applied to genes whose silencing interferes with plant regeneration or causes embryo lethality or severe pleiotropic phenotypes. In such cases the inducible RNAi vectors are used which can confer transient and local silencing. Ethanol- or estrogen-inducible vectors were developed for transient RNAi expression. In the case of an ethanol-inducible vector, a transcriptional regulator, AlcR, is constitutively expressed, and the RNAi cassette is inserted behind the alcA promoter. After ethanol treatment, AlcR binds to the alcA promoter ,and transcription of the downstream RNAi sequences is activated [54]. Also, a Cre/loxP-mediated recombination and a chemical inducing RNAi vector were developed for the stringent control of expression of an RNAi cassette. A chimeric transcriptional factor, XVE, was constitutively expressed which when binds to estrogen induces the transcription of a Cre recombinase gene. The resulting Cre recombinase removes a fragment that blocks transcription of an RNAi cassette [55]. Similarly, for controlled expression, the pOp6 promoter was used which could allow the rapid induction of RNAi across the whole plant or in limited tissues under investigation. The induction was controlled by the expression of a synthetic transcription factor, LhGR, which can bind to pOp6 in the presence of dexamethasone and initiate the transcription of downstream RNAi cassette [56]. The pHELLSGATE vector-based inducible RNAi vector is also available [57].
Improvement of hpRNA construct for efficient silencing led to the establishment of ihpRNA construct. Using hpRNA constructs containing sense/antisense arms ranging from 300 to 500 nt gave efficient silencing in a wide range of plant species, and inclusion of an intron in these constructs had a consistently enhancing effect. An intron-spliced hpRNA construct gives a higher proportion of silenced transformants than intron-free hpRNA constructs. The process of intron-splicing aligns the arms of the hpRNA, facilitating their duplex formation in the spliceosome complex, whereas the arms of hpRNAs have to find their self-complementarity by random tethered collisions. Intron-spliced hpRNA facilitates the more efficient and steady levels of duplex RNA formation that are sufficient for PTGS. Similarly, the tighter loop of ihpRNA gives more nuclease-stable and higher steady-state duplex RNA levels than the larger-looped hpRNA [36]. In plants, intron-containing hairpin RNA constructs with a spliceable intron as spacer sequence had the highest efficiency, with 80% to approximately 100% transformants showing silencing of target genes [10, 33, 36].
RNA interference has been used to develop efficient strategies to silence targeted genes in a wide range of species. Posttranscriptional silencing of plant genes using antisense or co-suppression constructs usually results in only a modest proportion of silenced individuals. Recent work has demonstrated the potential of the constructs encoding intron-containing self-complementary “hairpin” RNA to efficiently silence genes. The degree of silencing with these constructs was much greater than that obtained using either co-suppression or antisense constructs. Currently, the ihpRNA technology has become one of the most powerful tools for gene discovery and gene engineering in plants.
The worldwide spread and increasing transmission of diseases such as dengue, chikungunya, and Zika transmitted by
Different authors have coined the term “excito-repellency” to refer to the combined effect of escape responses after the contact with the insecticide-treated area [5, 8, 9]. Likewise, studies based on the excito-repellency test system have reported that the nutritional status and physiological conditions (including age) and the duration of the tests can significantly influence the response behavior to insecticides. Therefore, chronological age, physiological status, and innate (circadian) activity patterns should be carefully considered for the proper selection of mosquitoes used in the tests [3].
The behavior of mosquitoes in response to insecticides continues to be poorly studied and understood. They do not refer to anything other than the locomotor response of mosquitoes after capturing a stimulus, either contact “irritation” or noncontact spatial repellency “deterrence” resulting from the capturing of chemical emanations “odors” by nature and/or evaporation of chemical substances [10, 11]. On the other hand, understanding mosquito behavior will enable the selection of insecticides, the operational planning of cost-effective and long-term interventions and the development of innovative surveillance and vector control tools and strategies [12, 13].
Most of the interventions for
Environmental interventions (“Immature mosquito control”) are based on actions to eliminate breeding sites for these organisms, impacting adult populations. These interventions represent sustainable and safe methods as there are limited risks of environmental contamination and toxicity [18, 19]. Control of the immature stages of dengue vectors is generally conducted larval habitats using biological, chemical, environmental, or mechanical methods to maximize the reduction in vector population density [13]. The principal environmental methods are container covers with and without insecticides, waste management with and without direct garbage collection, elimination of breeding sites, drinking water supply, and urban planning [2, 19]. Similar to environmental interventions, mechanical and physical control, consists of cleaning breeding sites and physical barriers such as mosquito nets and curtains [15].
The biological control interventions to control mosquitos are based on pathogens and mosquito symbionts. Regarding successful experiences of biological control, some examples of interventions regarding the vertical or community-based introduction of Cyclopoid copepods (
Chemical control is an essential element in vector control strategies worldwide and can be implemented under two approaches: 1) to reduce density/increase vector mortality by adulticides and larvicides application and 2) to reduce human vector contact by insecticide-treated materials (ITMs) such as long-lasting insecticide nets (LLINs), traditional nets, and personal repellents [2]. Vector control programs have favored insecticides to control adults, mainly based on ultralow-volume (ULV) space application for outdoors in open spaces due to the ease of covering large areas in the shortest possible time and thermal fog indoors [26, 27, 28]. Aerial ultralow-volume (AULV) applications are also being tested in México [29]. Targeted indoor residual spraying (IRS, TIRS) is another control method with evidence of efficacy in México [30].
The most common insecticide products to control malaria and any other mosquito vector transmission combine two different modes of action: 1) conventional insecticide activity that kills mosquitoes exposed to the insecticide, and 2) deterring mosquitoes away from humans [17]. Space spraying of insecticides is still considered to be a valuable tool to control the vectors of human diseases [31]. S-methoprene, pyriproxyfen, temephos, and Bti are among larvicidal and pupicidal agents recommended and approved by the World Health Organization (WHO) to treat
Insecticide-treated materials (ITMs) can provide bite protection by killing or repelling vectors [6]. The use of nets impregnated with long-lasting insecticides used as pavilions or mosquito nets is currently one of the most promoted strategies to reduce the transmission of arboviral or parasitic diseases [17, 33]. Long-lasting insecticidal nets (LLINs) are materials pretreated with insecticides designed to prolong their useful life. Studies in several Latin American countries indicate that using LLINs fixed on doors and windows and insecticide-treated screening (ITS) are innovative approaches to control vector mosquito populations, and they may be promising in reducing the transmission of diseases such as dengue [34, 35, 36, 37]. The efficacy of these interventions is reflected in reduced vector-human interaction and sustained indoor adult vector densities as blood-fed and arbovirus-positive
Spatial repellents (SRs) are products containing volatile chemicals that disperse in the air under ambient conditions. Besides, the term “spatial repellency” is used here to refer to a range of insect behaviors induced by airborne chemicals that result in a reduction in human-vector contact and, therefore, personal protection. [42, 43]. The behaviors can include movement away from a chemical stimulus, interference with host detection (attraction inhibition), and feeding response [43]. In clinical trials, SR products reduced malaria and
For example, indoor residual spraying against malaria vectors depends on whether mosquitoes rest indoors (i.e., endophilic behavior); likewise, the optimum effectiveness of insecticide-treated nets presumably depends on vectors biting at hours when most people are in bed. Prospects for genetic control by sterile males or genes rendering mosquitoes harmless to humans will depend on competitive mating behavior [45]. Some works have provided evidence for the existence of behavioral modification because of widespread IRS or ITN use [46]. The behavioral responses of mosquitoes to insecticide products must be determinate to understand the main mechanisms involved in their effectiveness; this task is vital not only for success in chemical control strategies used in vector control programs but also for the development of new insecticides and the design of innovative control strategies [6, 12, 47]. The devices developed to analyze the behavior of mosquitoes evaluate three main components of chemical action: 1) contact irritation (excito-repellency), 2) spatial repellency, and 3) toxicity [8, 47, 48, 49].
Malaithong et al. [11] criticize the subjective way of using terms such as avoidance, excitation, irritability, deterrence, and excito-repellency by different authors in a system based on “success-repellency” tests, to refer to locomotor response behavior caused by contact with insecticides. The experts point out that this test system distinguishes two main locomotor responses: 1) stimulation of the musculoskeletal apparatus “excitation,” “irritation” as a result of tarsal contact with a treated surface and 2) noncontact spatial repellency “deterrence” as a result of chemical emanations, that is, the capture of “odors” in the evaporation phase of chemical compounds at a distance, that is, the capture of “odors” in the evaporation phase of chemical compounds at a distance, without the need to make contact with the treated area and emphasizes the need for a clearer understanding of the behavior, stimuli, effects, and mechanisms involved in the response locomotive of mosquitoes against different xenobiotics.
Kongmee [50] reported no repellency effect without contact generated by insecticides in a study with anophelines based on excite-repellency tests with two pyrethroids; nevertheless, deltamethrin produced a high irritating effect, while bifenthrin exhibited low levels. At operational field doses of alpha-cypermethrin, low noncontact repellency was observed in three mosquito populations; thus, spatial repellency may play a minor role in escaping vectors from treated surfaces, with contact irritation being the most important main effect on
The functions of insect chemoreceptors have primarily been studied using antennae (olfactory receptors) and mouthparts (gustatory receptors) through specialized structures called sensilla, which comprise neurons and non-neuronal support cells, extracellular lymph fluid, and a precisely shaped cuticle [54]. Other appendages with chemoreceptive sensilla include the leg tarsi and the anterior wing margin. Their specific roles in chemoreception and mosquito behavior remain largely unknown [55]. This review provides a brief description and illustration of the sensilla and its function in chemical stimulus, transduction, recognition, and understanding in insect behavior.
Modified cuticular structures are the basic sensory unit in insects; they are made up of four components: sensory neurons, a thermogenic cell (socket), a trichogenous cell that gives rise to the hair, and a thecogenous cell that surrounds and protects the axon terminal and provides it with ions and nutrients (Figure 1C), sensory neurons are bipolar, extending their dendrites from the cuticular portion and their axons toward the central nervous system (CNS). Due to their external morphology, they can be classified into trichoid, basiconic, placoid, styloconic, coeloconic, and bell-shaped [58].
Sensory receptors in mosquitoes, the sensilla and the stimulus. A) Sensory receptors on appendages based on Sparks et al. [
These structures, which are found on the surface of mosquito antennae and maxillary palps (olfactory sensilla), as well as mouthparts, tarsi, and wings (gustatory sensilla), play a major role in host detection and other sensory-mediated behaviors. (Figure 1A) [56, 59, 60]. In mosquitoes, the trichoid sensilla is the most common and abundant sensory structure. A single pore is typically observed at the distal end of a blunt tip. (Figure 1C) [60]. Bohbot et al. (2014) described the basiconica, chaetica, and campaniformia sensilla in
Highly sensitive units can react to an external stimulus, mainly of “gustatory” character in the mouthparts, legs, and ovipositor, or olfactory character, mainly in the antennae and palps. Zwonitzer (1962) [63] within this classification includes those known as “general chemical sensitivity”, which in turn respond to volatile materials classified as “distance chemical receptors” that are moderately sensitive. The categorization also comprises the receptors that are stimulated by nonvolatile substances (“chemoreceptors senses”), and those that are relatively insensitive and lead to protective responses defined as “general chemical senses.” Chemoreception is the ability to perceive specific chemical stimuli. It is one of the most evolutionarily ancient forms of interaction between living organisms and their environment; this physiological process occurs because of the contact with a chemical stimulus and presents a broad spectrum of sensations [64, 65].
The chemical signals produced by semiochemicals and chemical substances reach the sensilla and penetrate the cuticle through pores; these substances are hydrophobic and cannot pass through the fluid. Lymphatic (aqueous medium) is achieved thanks to odorant-binding proteins (OBPs) that encapsulate them and direct them to the surface of the sensory cell where a structural change of charges in the membrane causes the stimulus to be expelled on the nerve receptor (Figure 1D) [58].
Once the union between the stimulus and the receptor has occurred (stimulus-receptor complex), the event must be communicated to other parts of the sensory cell to ensure the final message through the effect of action potentials transmitted to the brain. The amplification involves a series of membrane-bound intracellular molecules, usually calcium; at least one ion channel detects the increase in calcium and opens, thus triggering membrane depolarization [58].
The sensory system acts as a filter allowing the insect to capture stimuli and to differentiate one stimulus from another. Therefore, receptor proteins and translation-associated molecules possess high specificity and sensitivity [66]. Several works have been published regarding the recognition by mosquitoes of emanations such as CO2 and lactic acid on human skin; these works also show that high concentrations of these compounds can have a deterrent effect on mosquitoes [67, 68].
In female and male
Different studies have compared the behavioral response of mosquitoes under different physiological conditions, including age. Experiments were carried out on
Oliver & Brooke [73] were the first to demonstrate the expression of resistance to insecticides because of multiple blood feedings; they also point out the variability of the expression levels of detoxification enzymes as a function of age that presented a decrease in these due to aging. This could be because blood feeding can modify the expression of genes that affect the action of detoxifying enzymes. This expression is more evident during the first, second, and third days following a single blood feeding showing a dependence on sex [74].
This association seems to influence mosquito susceptibility or resistance to insecticides in terms of mosquito parasitism. According to Agnew et al. [75], parasitism can act as a source modifying the costs of resistance to organophosphate insecticides and as qualitatively different interactions (increasing or decreasing relative fitness in resistant individuals) that occur depending on the type of resistance involved.
In several insect groups and disease vectors across the world, “physiological” resistance, metabolic, and target site modifications to insecticides have been well documented [76], including highly physiologically insecticide-resistant mosquitoes. It also implies the application of chemical products at higher concentrations, which is neither practically feasible nor cost-effective [12].
Although the behavioral responses of mosquito vectors to insecticides differ based on the type of product and concentration, it is also true that the product’s properties (irritant-repellent) can help to reduce human-vector interaction. The behavioral response of avoiding the treated surface seeks to integrate products with such properties to reduce the transmission of pathogens because they reduce the opportunity for blood feeding. Understanding mosquito behavior, including oviposition site selection, dispersal behavior, and competitive mating, can allow the development of innovative mosquito surveillance and control strategies to control these important and deadly insects better. On the other hand, molecular and structural studies and the signaling pathways of these receptors must be studied better to understand their function and role in resistance development.
“The authors declare no conflict of interest.”
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Thus proteomics, an area of research that detects all protein forms expressed in an organism, including splice isoforms and post-translational modifications, is more suitable than genomics for a comprehensive understanding of the biochemical processes that govern life. The most common proteomics applications are currently in the clinical field for the identification, in a variety of biological matrices, of biomarkers for diagnosis and therapeutic intervention of disorders. From the comparison of proteomic profiles of control and disease or different physiological states, which may emerge, changes in protein expression can provide new insights into the roles played by some proteins in human pathologies. Understanding how proteins function and interact with each other is another goal of proteomics that makes this approach even more intriguing. Specialized technology and expertise are required to assess the proteome of any biological sample. Currently, proteomics relies mainly on mass spectrometry (MS) combined with electrophoretic (1 or 2-DE-MS) and/or chromatographic techniques (LC-MS/MS). MS is an excellent tool that has gained popularity in proteomics because of its ability to gather a complex body of information such as cataloging protein expression, identifying protein modification sites, and defining protein interactions. 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