HERVs families associated with different diseases
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These books synthesize perspectives of renowned scientists from the world’s most prestigious institutions - from Fukushima Renewable Energy Institute in Japan to Stanford University in the United States, including Columbia University (US), University of Sidney (AU), University of Miami (USA), Cardiff University (UK), and many others.
\\n\\nThis collaboration embodied the true essence of Open Access by simplifying the approach to OA publishing for Academic editors and authors who contributed their research and allowed the new research to be made available free and open to anyone anywhere in the world.
\\n\\nTo celebrate the 50 books published, we have gathered them at one location - just one click away, so that you can easily browse the subjects of your interest, download the content directly, share it or read online.
\\n\\n\\n\\n\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
IntechOpen and Knowledge Unlatched formed a partnership to support researchers working in engineering sciences by enabling an easier approach to publishing Open Access content. Using the Knowledge Unlatched crowdfunding model to raise the publishing costs through libraries around the world, Open Access Publishing Fee (OAPF) was not required from the authors.
\n\nInitially, the partnership supported engineering research, but it soon grew to include physical and life sciences, attracting more researchers to the advantages of Open Access publishing.
\n\n\n\nThese books synthesize perspectives of renowned scientists from the world’s most prestigious institutions - from Fukushima Renewable Energy Institute in Japan to Stanford University in the United States, including Columbia University (US), University of Sidney (AU), University of Miami (USA), Cardiff University (UK), and many others.
\n\nThis collaboration embodied the true essence of Open Access by simplifying the approach to OA publishing for Academic editors and authors who contributed their research and allowed the new research to be made available free and open to anyone anywhere in the world.
\n\nTo celebrate the 50 books published, we have gathered them at one location - just one click away, so that you can easily browse the subjects of your interest, download the content directly, share it or read online.
\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:"6838",leadTitle:null,fullTitle:"Power Plants in the Industry",title:"Power Plants in the Industry",subtitle:null,reviewType:"peer-reviewed",abstract:'The main aim of this study is to present power plants for all fields of industry. The chapters collected in the book are contributions by invited researchers with long-standing experience in different research areas. I hope that the material presented here is understandable to a wide audience, not only energy and mechanical engineering specialists but also scientists from various disciplines. The book contains seven chapters in two sections: (1) "Power Plants & Technical Details" and (2) "Power Plants in Different Fuels". This book shows detailed and up-to-date evaluations in different areas and was written by academics with experience in their field. It is anticipated that this book will make a scientific contribution to power plant workers, researchers, academics, MSc and PhD students, and other scientists in both the present and future.',isbn:"978-1-78985-084-0",printIsbn:"978-1-78985-083-3",pdfIsbn:"978-1-83962-008-9",doi:"10.5772/intechopen.73347",price:119,priceEur:129,priceUsd:155,slug:"power-plants-in-the-industry",numberOfPages:148,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"5e647d27dab23e014dd8881ac3d5931c",bookSignature:"Tolga Taner",publishedDate:"February 6th 2019",coverURL:"https://cdn.intechopen.com/books/images_new/6838.jpg",numberOfDownloads:8297,numberOfWosCitations:1,numberOfCrossrefCitations:1,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:4,numberOfDimensionsCitationsByBook:0,hasAltmetrics:1,numberOfTotalCitations:6,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 21st 2018",dateEndSecondStepPublish:"April 11th 2018",dateEndThirdStepPublish:"June 10th 2018",dateEndFourthStepPublish:"August 29th 2018",dateEndFifthStepPublish:"October 28th 2018",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6,7",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"197240",title:"Associate Prof.",name:"Tolga",middleName:null,surname:"Taner",slug:"tolga-taner",fullName:"Tolga Taner",profilePictureURL:"https://mts.intechopen.com/storage/users/197240/images/system/197240.jpg",biography:"Dr. Tolga Taner is the head of the Department of Motor Vehicles and Transportation Technology at Aksaray University, Turkey. He received a BS in Mechanical Engineering in 1998; an MS in Mechanical Engineering from Pamukkale University, Denizli, Turkey, in 2002; and a Ph.D. in Mechanical Engineering from the Gazi University of Engineering Faculty, Ankara, Turkey, in 2013. In 2018, he received the title of Associate Professor from the Inter-University Council (UAK). He also worked as a part-time lecturer in the Department of Technical Programs, Middle East Technical University, Turkey, from 2003 to 2006. His current research interests include exergy, renewable energy, and PEM fuel cells. He has published many scientific and conference papers and books. In addition, Dr. Taner has been a manager and researcher for many research projects.",institutionString:"Aksaray University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"4",totalChapterViews:"0",totalEditedBooks:"5",institution:{name:"Aksaray University",institutionURL:null,country:{name:"Turkey"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"803",title:"Power Plant Engineering",slug:"power-plant-engineering"}],chapters:[{id:"65051",title:"Introductory Chapter: Application of Power Plants in the Industry",doi:"10.5772/intechopen.82406",slug:"introductory-chapter-application-of-power-plants-in-the-industry",totalDownloads:968,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:null,signatures:"Tolga Taner and Mecit Sivrioglu",downloadPdfUrl:"/chapter/pdf-download/65051",previewPdfUrl:"/chapter/pdf-preview/65051",authors:[{id:"197240",title:"Associate Prof.",name:"Tolga",surname:"Taner",slug:"tolga-taner",fullName:"Tolga Taner"}],corrections:null},{id:"64215",title:"Modernization and Development Scenarios of the Power Plants in the Present Energy Market Context",doi:"10.5772/intechopen.80887",slug:"modernization-and-development-scenarios-of-the-power-plants-in-the-present-energy-market-context",totalDownloads:938,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The aim of this study is to analyze several possible hypotheses for the development of a power plant in the context of the local electricity and heat market. The energy market may be constraint by the probable cessation of the activity of the most important customer in the industrial market, hereinafter referred to as “strategic customer.” Stopping its activity would deprive the power plant of an annual electricity supply of 28.02% of the electricity production estimated in the “continuity” hypothesis with the strategic customer, namely an annual heat of 78.19% of the heat output of the same alternative. It is very clear that stopping the activity of the company constitutes a disability for the power plant, whose magnitude has been highlighted by the comparative analysis of several possible scenarios of modernization, modernization-development, and development. As far as the electricity market is concerned, the power plant belongs to the area that is strongly equipped with sources of electricity production. With regard to the thermal energy market, the power plant, following the decoupling of the strategic customer, loses an important and stable consumer and stays only with the production of heat for heating the buildings and the preparation of domestic hot water for the residents of the city. The study’s result is the hierarchy of the proposed technical scenarios from the point of view of the necessary investment, the variation of the electricity tariff on the market, and the environmental impact.",signatures:"Adelaida Mihaela Duinea",downloadPdfUrl:"/chapter/pdf-download/64215",previewPdfUrl:"/chapter/pdf-preview/64215",authors:[{id:"254222",title:"Dr.",name:"Adelaida Mihaela",surname:"Duinea",slug:"adelaida-mihaela-duinea",fullName:"Adelaida Mihaela Duinea"}],corrections:null},{id:"63435",title:"Pipeline Health Monitoring to Optimise Plant Efficiency",doi:"10.5772/intechopen.80844",slug:"pipeline-health-monitoring-to-optimise-plant-efficiency",totalDownloads:1219,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"This chapter presents technological innovations that support asset integrity management—a crucial activity for optimising plant efficiency. In ageing thermal and geothermal power plants, critical assets such as steam piping are subject to high pressures and temperatures that accelerate damage mechanisms. Traditionally, the critical locations of these assets undergo routine inspection which is both costly and time consuming and affects the plant reliability and energy availability. There is an increasing trend in the application of non-destructive testing (NDT) and information technologies to in-service monitoring of these assets. The aim of this chapter is to provide a comprehensive overview of the state-of-the-art monitoring technologies for steamlines, with a focus on high temperature ultrasonic guided wave techniques. The enabling technologies, which include high temperature sensors, diagnostic data analysis algorithms and their monitoring performances, are reviewed. These technological advancements enable inspection without interruption of plant operations, and provide diagnosis and prognosis data for condition-based maintenance, increasing plant safety and its operational efficiency.",signatures:"Anurag Dhutti and Tat-Hean Gan",downloadPdfUrl:"/chapter/pdf-download/63435",previewPdfUrl:"/chapter/pdf-preview/63435",authors:[{id:"253101",title:"Dr.",name:"Tat-Hean",surname:"Gan",slug:"tat-hean-gan",fullName:"Tat-Hean Gan"},{id:"265553",title:"Mr.",name:"Anurag",surname:"Dhutti",slug:"anurag-dhutti",fullName:"Anurag Dhutti"}],corrections:null},{id:"63214",title:"Results of Full Scale Modeling of Electromagnetic Pulse Impact for Lightning Protection of Power Plants",doi:"10.5772/intechopen.80560",slug:"results-of-full-scale-modeling-of-electromagnetic-pulse-impact-for-lightning-protection-of-power-pla",totalDownloads:940,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:1,abstract:"This chapter summarizes the results of experimental modeling of lightning impacts that has been carried out several years on the problem of lightning protection of electric power objects, including power plants, primarily in order to increase the stability of their work. The main purpose of the research is to offer the testing facilities and testing schemes of lightning protection. A feature of the models proposed to the attention is the use of the energy of an explosive magnetic generator (EMG). In the first part of the chapter, the investigation connects with direct lightning current impact. For this purpose, a prototype of mobile testing complex on the basis of an explosive magnetic generator (MTC EMG) was developed. The results of MTC EMG field testing for loads with ohmic resistances of 2–10 Ω in the form of current and voltage pulses are presented. The results of an electromagnetic impulse impact in the near field of lightning were modeled experimentally in the second part of the chapter. As a result, the electrical field strength with a rising of voltage front about 100 ns were up to 500 kV/m, and about 0.2 T/μs of the magnetic induction increasing were obtained in the experiments. The paper provides estimates of the techno-economic analysis of the practical application of the development.",signatures:"Vladimir Fortov, Alexei Shurupov, Valentina Zavalova, Alexander\nKozlov, Mihail Shurupov and Nina Shurupova",downloadPdfUrl:"/chapter/pdf-download/63214",previewPdfUrl:"/chapter/pdf-preview/63214",authors:[{id:"253020",title:"Ph.D.",name:"Valentina",surname:"Zavalova",slug:"valentina-zavalova",fullName:"Valentina Zavalova"},{id:"253022",title:"Dr.",name:"Alexei",surname:"Shurupov",slug:"alexei-shurupov",fullName:"Alexei Shurupov"},{id:"267436",title:"Dr.",name:"Alexander",surname:"Kozlov",slug:"alexander-kozlov",fullName:"Alexander Kozlov"},{id:"267437",title:"Dr.",name:"Mihail",surname:"Shurupov",slug:"mihail-shurupov",fullName:"Mihail Shurupov"},{id:"267438",title:"M.Sc.",name:"Nina",surname:"Shurupova",slug:"nina-shurupova",fullName:"Nina Shurupova"},{id:"270584",title:"Associate Prof.",name:"Vladimir",surname:"Fortov",slug:"vladimir-fortov",fullName:"Vladimir Fortov"}],corrections:null},{id:"64317",title:"Hybrid Power Plants: A Case Study",doi:"10.5772/intechopen.80034",slug:"hybrid-power-plants-a-case-study",totalDownloads:1142,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Energy can be treated as an essential element for the development of society. Therefore, aspects like process’ efficiency and environmental impacts must be considered when choosing the supply source. In Brazil, an event showed the fragility of a system that relies on in only one source to attend their necessities; a truckers strike made the whole country stop. The energy sector has a similar situation; more than 60% of Brazilian energetic matrix is represented by one source, hydroelectric power plants. The availability of solar radiation and wind in Brazil makes it possible to diversify its energetic matrix. Thus, the aim of this study is investigating the potential of hybrid solar-wind power plants in two basins of Minas Gerais—Brazil, São Francisco Basin and Jequitinhonha Basin, as well as compare their viabilities in order to address social issues. By analyzing INMET database and economic factors, the study has shown that it is feasible to implement renewable power plants in the basins of the study area, whether individually (solar or wind energy) or hybrid system. It shows in addition that hybrid system should be prioritized, since it presents lower cost, when compared to solar power plant, and more reliability due to seasonality of both sources.",signatures:"Eduarda Moreira Nascimento, Júnio de Souza Damasceno and\nSabrinne Kelly Souza",downloadPdfUrl:"/chapter/pdf-download/64317",previewPdfUrl:"/chapter/pdf-preview/64317",authors:[{id:"252477",title:"Dr.",name:"Junio",surname:"Damasceno",slug:"junio-damasceno",fullName:"Junio Damasceno"},{id:"262354",title:"Ms.",name:"Sabrinne",surname:"Souza",slug:"sabrinne-souza",fullName:"Sabrinne Souza"},{id:"262363",title:"BSc.",name:"Eduarda",surname:"Nascimento",slug:"eduarda-nascimento",fullName:"Eduarda Nascimento"}],corrections:null},{id:"62970",title:"Nuclear Fusion Power Plants",doi:"10.5772/intechopen.80241",slug:"nuclear-fusion-power-plants",totalDownloads:1786,totalCrossrefCites:1,totalDimensionsCites:4,hasAltmetrics:1,abstract:"Nuclear fusion, the process that powers the sun and the stars, is heralded as the ultimate energy source for the future of mankind. The promise of nuclear fusion to provide clean and safe energy, while having abundant fuel resources continues to drive global research and development. However, the goal of reaching so-called “breakeven” energy conditions, whereby the energy produced from a fusion reaction is greater than the energy put in, is yet to be demonstrated. It is the role of ITER, an international collaborative experimental reactor, to achieve breakeven conditions and to demonstrate technologies that will allow fusion to be realized as a viable energy source. However, with significant delays and cost overruns to ITER, there has been increased interest in the development of other fusion reactor concepts, particularly by private-sector start-ups, all of which are exploring the possibility of an accelerated route to fusion. This chapter gives a comprehensive overview of nuclear fusion science, and provides an account of current approaches and their progress towards the realization of future fusion energy power plants. The range of technical issues, associated technology development challenges and future commercial opportunities are explored, with a focus on magnetic confinement approaches.",signatures:"Shutaro Takeda and Richard Pearson",downloadPdfUrl:"/chapter/pdf-download/62970",previewPdfUrl:"/chapter/pdf-preview/62970",authors:[{id:"251254",title:"Prof.",name:"Shutaro",surname:"Takeda",slug:"shutaro-takeda",fullName:"Shutaro Takeda"},{id:"262366",title:"Mr.",name:"Richard",surname:"Pearson",slug:"richard-pearson",fullName:"Richard Pearson"}],corrections:null},{id:"62660",title:"Flexible Operation of Supercritical Power Plant via Integration of Thermal Energy Storage",doi:"10.5772/intechopen.79735",slug:"flexible-operation-of-supercritical-power-plant-via-integration-of-thermal-energy-storage",totalDownloads:1306,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"This chapter presents the recent research on various strategies for power plant flexible operations to meet the requirements of load balance. The aim of this study is to investigate whether it is feasible to integrate the thermal energy storage (TES) with the thermal power plant steam-water cycle. Optional thermal charge and discharge locations in the cycle have been proposed and compared. Dynamic modeling and simulations have been carried out to demonstrate the capability of TES integration in supporting the flexible operation of the power plant. The simulation software named SimuEngine is adopted, and a 600 MW supercritical coal-fired power plant model is implemented onto the software platform. Three TES charging strategies and two TES discharging strategies are proposed and verified via the simulation platform. The simulation results show that it is feasible to extract steam from steam turbines to charge the TES and to discharge the stored thermal energy back to the power generation processes. The improved capability of the plant flexible operation is further studied in supporting the responses to the grid load demand changes. The results demonstrated that the TES integration has led to much faster and more flexible responses to the load demand changes.",signatures:"Decai Li, Wenbin Zhang and Jihong Wang",downloadPdfUrl:"/chapter/pdf-download/62660",previewPdfUrl:"/chapter/pdf-preview/62660",authors:[{id:"253189",title:"Prof.",name:"Jihong",surname:"Wang",slug:"jihong-wang",fullName:"Jihong Wang"},{id:"255747",title:"Mr.",name:"Decai",surname:"Li",slug:"decai-li",fullName:"Decai Li"},{id:"262087",title:"Dr.",name:"Wenbin",surname:"Zhang",slug:"wenbin-zhang",fullName:"Wenbin Zhang"}],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},subseries:null,tags:null},relatedBooks:[{type:"book",id:"6469",title:"Application of Exergy",subtitle:null,isOpenForSubmission:!1,hash:"0836749bb350a373d5e2628c73539698",slug:"application-of-exergy",bookSignature:"Tolga Taner",coverURL:"https://cdn.intechopen.com/books/images_new/6469.jpg",editedByType:"Edited by",editors:[{id:"197240",title:"Associate Prof.",name:"Tolga",surname:"Taner",slug:"tolga-taner",fullName:"Tolga Taner"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6285",title:"Proton Exchange Membrane Fuel Cell",subtitle:null,isOpenForSubmission:!1,hash:"ed010c881a38d577f89ccb714c17f785",slug:"proton-exchange-membrane-fuel-cell",bookSignature:"Tolga Taner",coverURL:"https://cdn.intechopen.com/books/images_new/6285.jpg",editedByType:"Edited by",editors:[{id:"197240",title:"Associate Prof.",name:"Tolga",surname:"Taner",slug:"tolga-taner",fullName:"Tolga Taner"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9385",title:"Renewable Energy",subtitle:"Technologies and Applications",isOpenForSubmission:!1,hash:"a6b446d19166f17f313008e6c056f3d8",slug:"renewable-energy-technologies-and-applications",bookSignature:"Tolga Taner, Archana Tiwari and Taha Selim Ustun",coverURL:"https://cdn.intechopen.com/books/images_new/9385.jpg",editedByType:"Edited by",editors:[{id:"197240",title:"Associate Prof.",name:"Tolga",surname:"Taner",slug:"tolga-taner",fullName:"Tolga Taner"}],equalEditorOne:{id:"186791",title:"Dr.",name:"Archana",middleName:null,surname:"Tiwari",slug:"archana-tiwari",fullName:"Archana Tiwari",profilePictureURL:"https://mts.intechopen.com/storage/users/186791/images/system/186791.jpg",biography:"Dr. Archana Tiwari is Associate Professor at Amity University, India. Her research interests include renewable sources of energy from microalgae and further utilizing the residual biomass for the generation of value-added products, bioremediation through microalgae and microbial consortium, antioxidative enzymes and stress, and nutraceuticals from microalgae. She has been working on algal biotechnology for the last two decades. She has published her research in many international journals and has authored many books and chapters with renowned publishing houses. She has also delivered talks as an invited speaker at many national and international conferences. Dr. Tiwari is the recipient of several awards including Researcher of the Year and Distinguished Scientist.",institutionString:"Amity University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"Amity University",institutionURL:null,country:{name:"India"}}},equalEditorTwo:{id:"197609",title:"Prof.",name:"Taha Selim",middleName:null,surname:"Ustun",slug:"taha-selim-ustun",fullName:"Taha Selim Ustun",profilePictureURL:"https://mts.intechopen.com/storage/users/197609/images/system/197609.jpeg",biography:"Dr. Taha Selim Ustun received a Ph.D. in Electrical Engineering from Victoria University, Melbourne, Australia. He is a researcher with the Fukushima Renewable Energy Institute, AIST (FREA), where he leads the Smart Grid Cybersecurity Laboratory. Prior to that, he was a faculty member with the School of Electrical and Computer Engineering, Carnegie Mellon University, Pittsburgh, PA, USA. His current research interests include power systems protection, communication in power networks, distributed generation, microgrids, electric vehicle integration, and cybersecurity in smart grids. He serves on the editorial boards of IEEE Access, IEEE Transactions on Industrial Informatics, Energies, Electronics, Electricity, World Electric Vehicle and Information journals. Dr. Ustun is a member of the IEEE 2004 and 2800, IEC Renewable Energy Management WG 8, and IEC TC 57 WG17. He has been invited to run specialist courses in Africa, India, and China. He has delivered talks for the Qatar Foundation, the World Energy Council, the Waterloo Global Science Initiative, and the European Union Energy Initiative (EUEI). 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\r\n\tThe study of global biogeochemical cycles of biogenic elements is necessary to describe the surrounding world, as well as to understand the trajectory of biosphere evolution and to realize the threats with which the humanity will have to face in the nearest and remote future. The book is devoted to the global biogeochemical cycles of biogenic elements: carbon, nitrogen, oxygen, sulfur, phosphorus and hydrogen. The participation of these elements in the synthesis of "living" matter on the Earth says by itself that biogeochemical cycles of biogenic elements are somehow linked and interdependent. To find out these links and to establish the key reactions of the cycles and coupling points are the most important tasks of researchers in order to get a precise and complete picture of the world. Recently obtained findings require generalization and comprehension and set the tasks of elucidating the interaction between the cycles, establishing the role and participation of the photosynthesis in the interaction, determining driving forces and building physic chemical models. This is pushed by deteriorating environmental conditions, global climate change and warming, ice melting and other problems.
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Their existence in the genome of mammals is only known since 1970 [2], although, they resulted from ancestral infections by exogenous retroviruses millions of years ago. During an infection, the exogenous retroviruses are able to integrate one copy of their genome (provirus) into the genome of the host. Thus, they can stay permanently associated with the host and be transmitted horizontally by the creation of new virions (the typical spread of an infectious virus). Only when they infect a germ line cell, the integrated DNA can become part of the gene pool and be transmitted in a Mendelian fashion like ERVs [1, 3-5], as shown in Figure 1. Those who are present in the human genome are named human endogenous retroviruses (HERVs).
The endogenization process profoundly impacts on the survival and evolution of the virus and the host. It results from the balance achieved between the immune surveillance and the virus virulence [6]. In this way, the HERVs must surpass the host’s antiviral defense mechanisms and infect the germ cells without causing a cytotoxicity that would prevent persistence in the progeny of the host [6]. Furthermore, from this moment on, all host cells are carriers of an integrated provirus [6].
The retroviral insertion is aleatory, in the sense that no specific sites for retroviral integration exist in the host genome. Nonetheless, due to the epigenetic chromatin packaging, integrated HERVs elements are more commonly found within the transcriptionally active genome [6]. Currently, HERVs comprise nearly 8% of the human genome [7], distributed in approximately 31 independently acquired multigene families [8]. Even though no standard nomenclature has been defined for HERVs, they have been classified based on their homology with different groups of exogenous retroviruses. They are grouped as class I, class II, or class III retroviruses considering their homology with Gamma and Epsilon retroviruses, Betaretrovirus or Spumavirus, respectively [9, 10]. The family name is usually given by “HERV” followed by a one-letter amino acid code that corresponds to the tRNA specific of the site used to initiate reverse transcription [10]; consequently, the HERV-W family would use a tryptophan.
As mentioned, HERVs have a similar structure to proviruses of infectious retroviruses, with three principal genes,
In addition to these “domestic HERVs,” several studies show reactivation of HERVs under pathologic conditions, such as different types of cancer [14-20]; autoimmune diseases including multiple sclerosis (MS) [21-37], rheumatoid arthritis (RA) [38], psoriasis [39], or systemic lupus erythematosus (SLE) [40]; and other diseases like schizophrenia [41, 42]. Nonetheless, we do not know whether their reactivation or increased expression is a causal effect, or conversely, is an underlying consequence of the disease.
Many factors can interfere or modulate the expression of HERVs, such as recombination events between two or more replication-defective HERVs [43, 44], infectious agents like Human herpesvirus 6 (HHV6) [34, 45] and Epstein–Barr virus (EBV) [46, 47], several transcription factors [31, 48], and the epigenomic context of the HERVs [6, 49, 50].
Recombination events
Two or more replication-defective HERVs can restore their own defects through recombination events, resulting in a replication-competent retrovirus [5]. Even though this is an infrequent event, a study in mice points to a significantly increased frequency in specific immune deficiencies [44]. Furthermore, it has been demonstrated that recombination between three HERV-K defective proviruses is possible, leading to an infectious retrovirus [5, 43].
Infectious agents
A putative explanation about the preferential expression of HERVs found in human brain samples could be the tropism of specific viruses and bacteria to the central nervous system (CNS). Neurotropic agents like herpesvirus [29, 51], Toxoplasma gondii [52], or certain strands of influenza virus [53] are able to cross the hematoencephalic barrier into the CNS. Usually, they are intercepted by cerebral macrophages leading to an abortive infection, but their transient presence in the CNS could activate the HERVs expression as a consequence of their immediate-early (IE) genes expression [4]. The expression of the IE genes of herpes simplex virus type 1 (HSV-1) and its interaction with the transcription factor binding sites situated in the U3 region of the LTR, such as AP-1 [54] and Oct-1 [55], lead to an activation of transcription in HERV-K and HERV-W families.
The herpesviruses are one of the best candidates: they may be neurotropic, remain latent, and can be reactivated. Furthermore, the expression of the Env epitopes in the surface of B cells and monocytes could be a consequence of the interaction between HERVs and herpesviruses [25]. Thus, the herpesviruses could play a dual role in neurodegenerative diseases, acting as pathological entities
Transcription factors
An important component of the antiviral innate immunity is the regulation of the expression and replication of HERVs by different transcription factors [48]. In HERV-W and HERV-K elements, both families previously related to multiple sclerosis (MS), different binding sites for transcription factors such as NF-Kβ [31, 48] are located in their promoter regions and could drive an increased expression of HERVs during inflammation.
Epigenomic context
The chromatin state as well as the methylation state of GpC islands within the HERV promoter and regulatory regions seem to be crucial factors in the control of HERVs expression [49, 50]. Both play an important role as a part of the defense system against the potential effects of inserted sequences. Previously published studies describe how proviruses and solitary LTRs are densely methylated under physiological conditions, but hypomethylated in placenta [49, 50]. Thus, DNA hypomethylation, as observed in certain types of cancer, could allow reactivation of retroelements. In MS, HERVs have been described as susceptible elements to undergo epigenetic modifications, mainly due to modifications in the methylation state, resulting in activation of their expression and, consequently, inappropriate activation of the immune system.
Even though most inserted copies in the human genome are defective copies, some HERVs could maintain the potential to cause or contribute to disease by different mechanisms [5]. As mentioned, HERVs may alter cellular functions by two ways, either acting as a genetic element or as a viral pathogen [6].
Gene disruption
HERVs, like transposons, are able to experience transposition, recombination, and integration cycles. Some HERVs families include a high number of copies in the genome. It is believed that these families have been spread around the genome through the reintegration of a provirus. However, each new integration process increases the risk of a harmful insertion. They can disrupt genes present in their integration sites, for example, HERV-K integrations have been identified into tumor suppressor genes like
Modulation of gene expression
Some HERVs conserve regulatory sequences that can operate as functional promoters, enhancers, or polyadenylation signals, so they could change the expression of adjacent or distal genes [4]. They can also form part of regulatory RNAs: microRNAs (miRNA), small interfering RNAs (siRNA), and long intergenic noncoding RNAs (lincRNAs), contributing to the complex regulatory network of gene expression [5]. Furthermore, HERVs integrated into introns can provide alternative transcription start and termination sites [5].
Pattern recognition receptors (PRRs)
The HERVs expression products, both nucleic acids and proteins, can modulate immune responses. They have the potential to interact with components involved in the immune innate response and to activate proinflammatory signaling pathways [57, 58]. Therefore, certain HERVs proteins could directly interact with specific toll-like receptors (TLRs), for example with TLR4, resulting in the production of TNFα and proinflammatory cytokines [58-60]. The nucleic acids derived from HERVs may also activate cytosolic PRRs; in this way, both an increased expression of RNA and the presence of cDNA in a nonfamilial compartment like the cytosol could activate PRRs [60]. Nonetheless, the human being has coevolved with endogenous retroelements and this could have shaped the sensibility of DNA sensors of the innate immune system, leading to an increased cDNA detection threshold to avoid an immune response against them. The cDNA levels are restrained by the action of gene products like Trex1 or SAMHD1 [60] and a loss-of-function mutation in these enzymes could result in the cDNA accumulation and the consequent sensors activation. This process would lead to a chronic immune response with release of pathogenic type I IFN and inflammatory mediators, similar to those observed in autoimmune diseases [60].
Viral proteins: molecular mimicry, superantigen activity, or immunosuppressive proteins
HERVs proteins hold epitopes to B and T cells and molecular mimicry between viral proteins and certain autoantigens may exist, resulting in an autoimmune response. Moreover, some HERVs sequences are able to encode for superantigens. Superantigens combine with MHC class-II molecules to form ligands that stimulate T cells [61], and this may end in an abnormal activation of autoreactive T lymphocytes [62].
Alternatively, evidences exist of the immunosuppressive activity of certain HERVs Env proteins [63, 64]. This activity is reminiscent of their exogenous antecessors, which in this way increased the viability of the virions in the host. This capacity has suffered an adaptation process, and nowadays it might be implicated in the materno-fetal tolerance and could also prevent the immune response to exogen pathogens and tumors [60].
Retroviral help for B cells
HERVs can also help B cells to quickly produce antibodies directed against pathogenic antigens [65]. The bacterial polysaccharide antigens and the carbohydrates linked to viral glycoproteins have the ability to stimulate B cells in the absence of T-cell help. These antigens are called thymus-independent antigens (TI), and they can be classified into two types: TI-1 or TI-2 antigens. TI-2 antigens cause extensive cross-linking of the BCR, leading to a quick differentiation of B cells into plasma cells. Finally, these plasma cells secrete protective antibodies, IgM and IgG [66]. However, the mechanism by which the TI-2 antigens activate B cells in the marginal zone without the help of T cells still remains poorly understood. It has been recently described that the cross-linking of B cells activates a signaling cascade, including the Bruton Tyrosine Kinase and the nuclear transcription factor NF-Kβ, allowing transcription of endogenous retroviral DNA [66]. The retroviral RNA may activate B cells by two complementary but different pathways: first, it could activate the retinoic-acid-inducible gene 1 receptor (RIG-1), resulting in a mitochondrial antiviral-signaling (MAVS); second, the RNA can be converted into DNA and can activate the cyclic GMP-AMP synthase (cGAS, cGAMP synthase). Finally, both signaling pathways would finish in the antigen-specific B-cell activation [65, 66].
HERVs represent the immunological limit between the self and the foreign. Their peculiar origin is very different from that of other genome elements, as they can share properties with infectious agents. Indeed, in case they would produce particles, these would not be so different from those originated from exogenous retroviruses. Therefore, they could activate the immune system and would induce autoimmunity [67]. As it has been previously discussed, HERVs have been associated, among other infectious or neurologic diseases, with different autoimmune diseases like MS [21-37], RA [38], psoriasis [39], T1D [68], or SLE [40], as shown in Table 1. Genome-wide association studies (GWAS) showed the existence of a genetic basis shared between different autoimmune diseases, discovering new immunogenic mechanisms implicated, and HERVs could be part of these shared genetic elements.
\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t
I | \n\t\t\tHERV-W | \n\t\t\tTrp | \n\t\t\tMS Schizophrenia HIV Osteoarthitis | \n\t\t\tHerpesviruses Transcription factors Toxoplasma gondii Influenza A virus | \n\t\t\tPro-inflammatory Env protein Superantigen activity OPCs differentiation interference Altered glial function | \n\t\t
I | \n\t\t\tHERV-F | \n\t\t\tPhe | \n\t\t\tMS | \n\t\t\tDemethylating agents | \n\t\t\tSuperantigen activity | \n\t\t
I | \n\t\t\tHERV-H | \n\t\t\tHis | \n\t\t\t3q13.31 microdeletion syndrome | \n\t\t\tN/A | \n\t\t\tGenetic deletion by recombination | \n\t\t
I | \n\t\t\tHERV-E | \n\t\t\tGlu | \n\t\t\tSLE | \n\t\t\tHypomethylation | \n\t\t\tImmunosupressor potential of Env | \n\t\t
I | \n\t\t\tHERV-P | \n\t\t\tPro | \n\t\t\tCancer | \n\t\t\tUnkown | \n\t\t\tUnknown | \n\t\t
II | \n\t\t\tHERV-K | \n\t\t\tLys | \n\t\t\tMS ALS HIV Schizophrenia T1D RA Juvenil arthitis Cancer | \n\t\t\tHerpersviruses HTLV-1 Type 1 IFN Transcription factors Hypomethylation | \n\t\t\tSuperantigen activity Neoepitopes Genes disruption | \n\t\t
HERVs families associated with different diseases
MS, multiple sclerosis; HIV, human immunodeficiency virus; SLE, Systemic lupus erythematosus; ALS, amyotrophic lateral sclerosis; T1D, type 1 diabetes; RA, rheumatoid arthritis, HTLV-1, human T-lymphotropic virus-1; IFN, interferon; Env, envelope; OPCs, olygodendrocyte precursor cells; N/A, not applicable. Based on Douville and Nath, 2014 [6].
MS is one of the conditions more frequently related with HERVs. It is a chronic progressive disease characterized by neuroinflammation in the CNS accompanied by demyelination, axonal damage, and progressive neurologic dysfunction [69]. It is a complex disease, originated from the interaction of genetic, environmental, and epigenetic factors [70]. Recently, its incidence seems to be increased; at present MS affects 2.3 million people in the world [71]. However, many aspects of its pathogenesis are still poorly understood. GWAS have not completely explained the MS genetic background [72-74], albeit including the ImmunoChip Project [75] a total of 110 single nucleotide polymorphisms (SNPs) have been associated with MS susceptibility. Even considering the strongest risk factor, the HLA-DRB1*15:01 allele, each SNP has a modest effect and all together are able to explain only 20–28% of MS heritability [75]. Part of the missing heritability could reside on HERVs, as repetitive regions were not analyzed in the GWAS. Those repetitive regions were previously considered as “junk DNA” because it was thought that they had little or no physiological role. However, nowadays we know that these sequences could play an important role in the development of autoimmune diseases, including MS.
In 1989, Perron et al. [76] described the presence of extracellular virions associated with reverse transcriptase activity in a culture of leptomeningeal cells (LM7) obtained from the cerebrospinal fluid (CSF) of an MS patient. In the beginning, it was thought that those virions could correspond to the human T-lymphotropic virus (HTLV-1) due to the similarities between the tropical spastic paraparesis (a demyelinating progressive disease) caused by HTLV-1, and MS. However, a new retroviral element called MSRV (Multiple-sclerosis-associated retrovirus) was identified, the founder of the HERV-W family [77]. This multicopy family, consisting of approximately 650 loci around the human genome [35], comprises a total of 311 inserts (more or less complete proviruses or pseudogenes) and 343 additional HERV-W LTRs [78].
Only the
Recently, a genetic screening was performed by specific PCR amplification followed by High Resolution Melting (HRM) analyses of the two MSRV-like env copies which show the ORF with the highest length similarity and homology to Syncytin (1614 bp), inserted in chromosome X (1428 bp) and in chromosome 20 (1419 bp). Both chromosomal origins show similar lengths of their respective ORFs, 10% shorter than the one measured for Syncytin, and could putatively originate functional proteins. The results pointed to the insertion in chromosome X, and not the one in chromosome 20, as an origin of MSRV. One polymorphism identified in chromosome X, rs6622139*T, was associated in women with MS susceptibility and severity [82], and it was also associated with higher MSRV-like
Since it was described, several studies have associated the HERV-W family with MS: the presence of MSRV-type Env protein has been found in demyelinated acute lesions in MS patients [31], as well as an increased number of DNA copies [84] or a higher prevalence of MSRV-type RNA in serum and CSF of MS patients compared with patients suffering from other neurological diseases or healthy controls from all ethnic groups [24, 27, 28, 31, 84-86]. The MSRV presence in serum and CSF is correlated with the clinical progression, severity, and prognosis of MS [28, 46], while the absence of MSRV relates with a more stable course of the disease [28, 36]. The MSRV production is stimulated by cytokines like TNFα, IL6, and IFNγ[87], and current MS therapies like IFN-β and Natalizumab, which are able to reduce MS symptomatology, promote a diminution of MSRV virus load levels in blood [87-89].
HERV-W Env proteins, MSRV-type Env, and Syncytin have proinflammatory and superantigenic properties. They can cause neuroinflammation, neurodegeneration, immune system dysregulation, and endoplasmic reticulum stress [4, 21, 22, 58, 90, 91]. Their pathogenicity has been studied
\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t
T lymphocytes | \n\t\t\tTCR | \n\t\t\tSuperantigen activity, T lymphocytes proliferation and CK liberation | \n\t\t
APC | \n\t\t\tTLR4 | \n\t\t\t|
HCMEC/D3 endothelial cell line | \n\t\t\tTLR4 | \n\t\t\tIL6, IL8 expression | \n\t\t
OPC | \n\t\t\tTLR4 | \n\t\t\tOPCs differentiation interference ( | \n\t\t
Known pathogenic mechanisms of MSRV
APC, antigen presenting cell; OPC, olygodendrocyte precursor cell; TCR, T-cell receptor; TLR4, toll-like receptor 4
Even though the HERV-W family is one of the HERV families more related to MS, other families like HERV-K18 [37, 93] or HERV-Fc1 [29, 94, 95] have also been associated with MS susceptibility.
HERV-K is a multicopy family including approximately 332 copies dispersed through the human genome. It is the only known retroviral element that codes for all the structural and enzymatic proteins (Gag, Prt, Pol), as well as for the Env protein and for the accessory Rec protein [96]. This family has been related with different autoimmune diseases as MS [37], type-1diabetes (T1D) [68], or juvenile rheumatoid arthritis [38]; and different cancer types [14-17]. One specific member of this family, HERV-K18, has been associated with MS susceptibility and its expression is induced by herpesvirus [97, 98] and by EBV [99-102], both viruses previously proposed as potential environmental factors involved in MS development [45, 51, 97, 103-108]. Three different variants of the HERV-K18 copy mapping to chromosome 1 [37] have been described. They conform haplotypes within the first intron of CD48 that can be defined by two SNPs (18.1 SNP1*A/SNP2*A, 18.2 SNP1*G/SNP2*G, 18.3 SNP1*A/SNP2*G), all of them coding for an Env protein with superantigenic properties. However, only one of these variants (18.3) has been associated with a higher risk to MS [37] and with an overall higher susceptibility to autoimmune diseases, as described by a meta-analysis including a total of 2656 patients and 2016 controls [93].
Considering the HERV-Fc family, a total of 6 HERV-Fc elements and 11 LTRs have been identified across the human genome. Among them, only two elements correspond to a complete HERV-Fc provirus (Fc1env and Fc2master) [109]. Related to MS, it has been observed that the HERV-Fc1 RNA levels were significantly increased in the plasma of patients suffering from active MS, compared to nonactive MS or controls [30]. The HERV-Fc1 is an unusual provirus, because it includes a single copy in the genome, located on Xq21.33. Furthermore, it is a recent acquisition for the genome, only present in humans, chimpanzees, and gorillas [109]. Nexo et al. [94] were the first to describe that rs391745, located in the promoter region of HERV-Fc1, was associated with MS susceptibility in Danish cohorts and, then, a replication study was performed with a Norwegian cohort. The latter study also detailed that the association was only observed in the nonprimary-progressive MS forms [29], results validated in further studies [95]. Regarding the HERV-Fc1 expression mechanisms, it has been observed that the transcriptional expression levels of HERV-Fc1 RNA sequences are negatively correlated with the methylation levels of CpG islands on the 5’ LTR region and, therefore, a higher HERV-Fc1 expression involves DNA demethylation [11, 110].
An increased expression of HERVs in several autoimmune diseases [21-40, 68] and different types of cancer [14-20], along with the decreased expression levels observed in successfully treated patients with immunomodulatory therapies [88, 89] or chemotherapy [111] point to the potential pathogenic role of HERVs and their putative consideration as a good target for new treatments.
A humanized monoclonal antibody anti-Env-SU MSRV/HERV-W, GNbAC1, has been studied as a putative MS treatment due to its potential neuroprotector effects [112-115]. The results of a phase IIa clinical trial [114] show that the GNbAC1 treatment blocks the transcription of proinflammatory genes mediated by Env, prevents the formation of nitrosantine, and restores OPC differentiation. Furthermore, GNbAC1 has advantages compared to other MS treatments, because the patients retain all their immune capacity. This treatment has also been studied in other diseases like diabetes and schizophrenia.
The proteins encoded by HERV-K
In addition to autoimmune diseases and cancer, the human immunodeficiency virus (HIV) has been also related to HERVs, particularly with the HERV-K family, raising the issue of potential beneficial effects of a therapy directed against HERVs in AIDS. Some studies report an increased expression of HERV-K provirus in HIV patients compared to controls [117, 118] and show that the immune responses against HERV-K decrease the HIV-1 viral load.
This work aimed to provide a systematic revision of HERVs, with particular emphasis on their potential pathogenic role in MS. Although many aspects of the etiology of this disease remain to be solved, different works support the relevance that HERVs may have in the etiopathogenesis of autoimmune diseases, and specifically in MS. HERVs may contribute to both, disease onset and maintenance, through an exacerbated activation of the immune system. Recently, the results of a phase IIa clinical trial that studies the effectiveness of a human monoclonal antibody (GNbAC1) as a therapeutic target in MS have been published with promising outcome. Thus, evidences support the role of HERVs as potential therapeutic armory in different autoimmune diseases, cancer, and HIV.
Crystallization plays an important role in the manufacture and purification of small-molecule active pharmaceutical ingredients (APIs). It is estimated that between 70 and 80% of all small-molecule, APIs have at least one crystallization step in their manufacturing processes [1, 2]. To facilitate other downstream processes, such as filtration, drying, dissolution testing, and formulation, it is often desirable to be able to consistently produce crystals with specific properties, such as crystal size distribution, crystal shape (habit), and polymorphic form. Such control over the crystallization process requires accurate descriptions of crystal nucleation and growth kinetics, as well as solubility, breakage, and agglomeration data [3]. Crystallization control has gained even more interest since the release of the United States Food and Drug Administration’s (FDA) Process Analytical Technology (PAT) framework, which aims to improve efficiency in pharmaceutical development, manufacturing, and quality assurance through innovative process development, analysis, and control [4].
Designing and implementing controlled crystallization processes can be time-consuming, and usually involve trial-and-error, model-based, or model-free (direct-design) approaches [5]. Accurate kinetic descriptions of crystal nucleation and growth form the backbone of the model-based approach which, although more complex, has the potential to alleviate the labor-intensiveness and facilitate the optimization of the traditional trial-and-error approach [6]. The models used in the model-based approach can even be used together, to more accurately describe the underlying crystallization mechanism(s). For example, in a study by Quilló and coworkers [6], three different crystal growth models were fit to experimental data, to find the most likely crystallization mechanism. They found that the underlying crystallization mechanism could best be described by an additive combination of simultaneous Birth-and-Spread (B+S) and Burton-Cabrera-Frank (BCF) models, which will be discussed later in this chapter. An example of the model-free approach is direct nucleation control (DNC), which attempts to directly control the number of nuclei present in a system. Using DNC, Abu Bakar and coworkers [5] were able to produce glycine crystals from water–ethanol mixtures that were larger than those obtained from uncontrolled crystallization.
To implement the above-mentioned approaches, the investigator needs to be able to monitor the crystallization process. The instrumental technique used will depend on what aspect of the process one needs to monitor. For the model-based approach, one would typically be interested in monitoring the supersaturation decay during crystallization. This can be done using techniques, such as attenuated total reflectance-Fourier transform infrared (ATR-FTIR) or near infrared (NIR) [7]. If the goal is to monitor the evolution of crystal size distribution over time, for example in the DNC study mentioned above, a technique like focused beam reflectance measurement (FBRM) can be used [5, 7]. In most investigations, the above-mentioned instrumental techniques are used concurrently, to gain as much information about the crystallization process as possible.
The crystallization processes and monitoring techniques mentioned above are concerned with crystallization from solution, where a reduction in temperature and/or the addition of an antisolvent is used to create supersaturated conditions. Under supersaturated conditions, the solute molecules can come together to form small masses, or nuclei, through a process called nucleation. The addition of more solute molecules to these nuclei is called crystal growth. There is also another type of crystallization that is of particular importance in pharmaceutical sciences, and that is crystallization from the amorphous state.
In this chapter, we will discuss the theoretical background of crystal nucleation and growth, look at examples of how these theoretical models can be implemented practically and give examples of applications in pharmaceutical research. We will also look at crystallization from the amorphous state and discuss techniques that can be used to delay it.
The thermodynamic driving force behind crystal nucleation and growth rates is a chemical potential difference (Eq. (1)):
where
The value of
Because the activity coefficients still need to be estimated, many studies make the simplifying assumption of setting the activity coefficients in Eq. (2) equal to 1, thereby expressing the supersaturation driving force only as a mole fraction
When dealing with phase equilibria, the traditional thermodynamic reference is a supercooled melt of the pure solute compound. Using this reference, the activity of a solute can be obtained from the enthalpy of fusion at the temperature of interest. In its most general form, this is done through Eq. (3) [14]:
where
A practical problem with Eq. (3) is that
A common assumption is to completely ignore
However, when working with temperature ranges normally used in pharmaceutical processes, some approximations obtained from Eq. (5) have shown to be inaccurate [15, 16].
Another common assumption is that
If we can experimentally determine the isobaric heat capacity of the solid,
Illustration of the experimental heat capacities (solid lines) and extrapolated heat capacities (dashed lines) used to approximate the heat capacity term in
If we have enough isobaric heat capacity data above the melting temperature, we can extrapolate down from
where
Once the values of
In the introduction, we mentioned that crystallization consists of two processes, namely nucleation and growth. Depending on the rates of these two processes, the molecules that make up the crystal may pack differently, giving rise to different crystal structures, or polymorphic forms, of the same compound.
Nucleation is the first step in the crystallization process, and it consists of two mechanisms, namely homogeneous and heterogeneous nucleation. Homogeneous nucleation is triggered by spontaneous fluctuations in the density of the liquid, while heterogeneous nucleation is triggered by contact of the liquid with a foreign solid surface, like an impurity or metastable polymorph [7]. Since most industrial crystallization processes involve heterogeneous nucleation, we will focus mainly on this mechanism, which can be expressed mathematically as Eq. (10):
where
where
Looking at Eq. (10), we see some interesting mechanistic features of the nucleation step. First off, we can expect a higher number of nuclei to form from solvents in which the solute is more soluble because these will give higher values of
where
In the introduction, we mentioned that there are model-free approaches to controlling crystallization and that DNC is an example of such an approach. In short, the DNC approach attempts to control the number of nuclei present by adding solvent or antisolvent, increasing or decreasing the temperature, or a combination of both, to manipulate the solute’s solubility. Abu Bakar and coworkers employed DNC and were able to produce progressively larger glycine crystals if they lowered the number of newly formed particles [5].
Another example of where the nucleation step of crystallization was used as an intervention point to control the crystal size distribution, is in the work of Fujiwara and coworkers [20]. Paracetamol (acetaminophen), like many other small-molecule APIs, tends to form agglomerates during crystallization, especially if the crystals are small (< 100
Illustration of how the degree of supersaturation (Run 2) was controlled to fall between the metastable limit (Meta) and the solubility curve (Csat) during cooling. Reproduced from ref. [
Once the nucleation step of crystallization is underway, more solute molecules can become incorporated into these nuclei and crystal growth can begin. Generally, crystal growth is considered to consist of two steps in series, namely volume diffusion and surface integration. Volume diffusion consists of the diffusion of solute molecules from the solution, through the boundary layer surrounding the crystal, to its surface. Surface integration can be split into three subprocesses, starting with the desolvation of a solute molecule unto the surface of the crystal, followed by the transfer of this molecule from its point of arrival across the crystal’s surface. The latter is referred to as surface diffusion. Finally, the desolvated molecule reaches an energetically favorable location (kink site) on the crystal’s surface and becomes incorporated into the crystal lattice [13]. The steps in crystal growth described above are illustrated in Figure 3.
A schematic representation of crystal growth. The steps are (1) the diffusion of a molecule from the solution to the crystal surface, (2) desolvation of the solute molecule unto the crystal surface, (3) transfer across the surface, and (4) incorporation of the molecule into the lattice. The numbering represents—(a) the terrace, (b) the step, and (c) the kink site. Reproduced from ref. [
In this section, we will discuss the different crystal growth mechanisms, their theoretical basis, and examples of how they are used in pharmaceutical sciences. The topic of surface energy models is quite extensive and will not be covered in detail in this section. However, it plays an important role in how different crystal growth rates can lead to different crystal habits and is, therefore, a relevant topic in solid-state pharmaceutical investigations, as it influences downstream processes, such as dissolution, powder flow, milling, granulation, and compaction. Therefore, we will start this section with a discussion on the Gibbs–Curie–Wulff theorem and then look at a popular surface energy model, namely the Bravais–Friedel–Donnay–Harker (BFDH) model.
Also, in this section, we will not discuss adsorption layer or diffusion-reaction models but will instead focus on the more popular power law, birth-and-spread, and screw dislocation models, and show how information regarding the rate-limiting steps in crystal growth can be inferred from the power-law model.
The Gibbs–Curie–Wulff theorem originated when Gibbs postulated that, at equilibrium, a crystal should take on a form such that the product of its surface area and the surface-free energy is minimized. Later, Curie proposed that there exists a direct proportionality between the normal growth rates of crystal faces and surface-free energy. Wulff later stated that, at equilibrium, there exists a central point (the Wulff point) within a crystal such that the distances of the crystal faces from this point are proportional to the specific surface-free energies of those faces [22].
Li and coworkers [22] investigated whether organic crystals, with their diverse molecular shapes, noncovalent interactions, and hydrogen bonding, would still follow the Gibbs–Curie–Wulff theorem. Using a heteroacene molecule as crystal former, they found that not only did the shape and shape evolution of their equilibrium crystals follow the Gibbs–Curie–Wulff theorem, but that the packing of the molecules inside the crystals also reflected the principle of minimizing surface-free energy.
A crystal lattice can be thought of as a repeating 3D pattern consisting of a set of points such that each point has identical surroundings. A lattice has three spatial dimensions (
And for a more complex tetragonal system [23, 24]:
The BFDH model attempts to predict the crystal habit based on the growth rates of the crystal facets. Concretely, the BFDH model states that the growth rate of a facet is inversely proportional to that facet’s interplanar spacing [21, 25]. Unlike the other crystal growth models that will be discussed later in this chapter, the BFDH model does not have a mathematical representation. Instead, predictions of crystal morphology based on the BFDH model are obtained from computer modeling software, such as Mercury 2020.1, Cerius2, or Materials Studio.
Since the BFDH model does not explicitly take into account factors, such as molecular interactions but is instead concerned with the geometric aspects of the crystal, there have been instances where predictions made from the BFDH model were not in agreement with experimental observations. For example, while using the BFDH model to predict the morphologies of two carvedilol polymorphs, Prado and coworkers found that their BFDH model predicted thicker crystals of form II than were experimentally obtained and the model also predicted more faces for form III [26]. Similarly, Nichols and Frampton found that the BFDH model predicted morphologies for paracetamol polymorphic forms I and II that differed significantly from those observed experimentally [27]. Instead of using the BFDH model to predict morphologies, Turner and coworkers [28] used it to identify the important morphological faces of lovastatin crystals obtained from different solvents. They found that the calculated attachment energies of these important faces correlated reasonably well with the observed crystal morphologies.
The first mathematical model of crystal growth we will discuss is the power-law, or empirical, model. As the name suggests it is an empirical model, raised to some power, and as such it does not offer much insight into the actual growth mechanism. However, it is widely regarded for its flexibility and for being less complex and easier to use than the birth-and-spread or screw dislocation models.
Concretely, the power-law model expresses the growth rate
where the kinetics constant,
and where
We mentioned that the power-law model does not explicitly offer insights into the specific crystal growth mechanism. However, recently certain consistencies between the values of
We also mentioned the flexibility of the power-law model. In a paper concerning the crystal growth kinetics of piracetam polymorphs, Soto and Rasmuson [13] substituted the supersaturation driving force in Eq. (15) with a different driving force, namely a mass balance expression based on Haüy’s law, which states that one can assume a crystal’s shape to be constant if nucleation, agglomeration, breakage and growth rate dispersion can all be assumed to be negligible. The driving force can then be expressed as Eq. (17):
where
where
The birth-and-spread model (B+S) models surface, or two-dimensional, nucleation which is believed to occur at intermediate levels of supersaturation. Under such conditions, surface nuclei can form at the edges, corners, and even faces of crystals, creating new growth steps [13]. These surface nuclei then spread to create a new crystalline layer. The B+S model is presented here in Eq. (19)–(22):
where
Despite being a very comprehensive model, there is a specific inconsistency between predicted data and experimental observation that should be pointed out, and that is that once growth from a surface nucleation site has spread all the way to the edge of the crystal face, forming a new crystalline layer, any further growth would require adsorption to a smooth surface. Since this kind of adsorption is energetically less favored than binding to dislocations, it would require high levels of supersaturation to overcome the energy barrier. However, experimental data have shown that crystal growth can occur at much lower degrees of supersaturation [8]. To address this inconsistency, a screw dislocation model can be used, which assumes that screw dislocations on a crystal surface serve as self-perpetuating growth sites.
The Burton–Cabrera–Frank (BCF) model [29] is a screw dislocation model, that assumes that crystal growth stems from screw dislocations on a crystal surface and that growth from these ledges leads to more ledges in a self-perpetuating fashion, thereby attempting to explain how crystal growth can be obtained from supersaturations lower than that needed for the mechanism described by the B+S model. The BCF model can be expressed mathematically as Eqs. (23)–(25):
where
Now that we have presented the B+S and BCF models, we can appreciate the simplicity of the power-law model. In the introduction, we mentioned that Quilló and coworkers [6] used the power law, B+S, and BCF models, separately and in different combinations, to model the crystal growth kinetics of a proprietary API and thereby attempted to ascertain the underlying growth mechanism(s). During the combination tests, they also tested for dominance of one model over the other, sudden mechanism crossover, and simultaneous mechanisms. They found that the growth process could be best described if both the B+S and BCF models were simultaneously active, in an additive manner, with a smooth crossover from initial surface nucleation dominated growth to screw dislocation (spiral growth) dominated growth.
One of the ways to improve the solubility of a drug is to prepare an amorphous solid from it. The molecules inside these amorphous solids lack the long-range ordered packing of their crystalline counterparts, giving the amorphous solids higher thermodynamic activity and generally enhanced solubility. Methods to enhance the solubility of drugs have become a hot topic over recent years, and will likely remain a topic of interest, especially considering that approximately 36% of the 698 drugs currently available as immediate-release oral preparations and 60% of the 28912 new APIs under development are classified as biopharmaceutics classification system (BCS) class II or IV drugs [30, 31]. The BCS attempts to classify a drug into one of four categories, depending on its aqueous solubility and membrane permeability [32]. BCS class II and IV drugs have poor aqueous solubility but good membrane permeability, and poor aqueous solubility and poor membrane permeability, respectively. In other words, the market share of poorly soluble drugs is likely to increase in the future.
Currently, there are a limited number of amorphous preparations on the market [33]. A specific cause for concern is that the high thermodynamic activity of an amorphous solid, responsible for its enhanced solubility, also makes it thermodynamically unstable and likely to convert back to a more stable, but less soluble, crystalline form. A mechanistic understanding of this conversion and the resulting glass-to-crystal (GC) growth is an important topic in pharmaceutical sciences.
GC growth rates cannot be readily explained by the thermodynamic driving force responsible for crystallization from solution, as described previously [34]. Initially, it was believed that crystallization from the amorphous state was dependent on the storage conditions, such as the storage temperature relative to the glass transition temperature
Both the surface and bulk crystallization processes are believed to be controlled by the self-diffusion of the molecules in the amorphous solid. Using indomethacin as model API, both Wu and Yu [38] and Swallen and Ediger [39] found that the crystal growth rates in the interior of their amorphous solids were proportional to the self-diffusion coefficients for temperatures close to
With the mechanism of surface crystallization understood, immobilization of the molecules at the surface makes for an appealing target to delay the crystallization process. Yu and coworkers [41] coated indomethacin glasses with gold (10 nm) and two polyelectrolytes. They found that even a single layer of polyelectrolyte was enough to inhibit the growth of exiting crystals and that the molecular mobility of molecules at the surface of an amorphous solid can be sufficiently suppressed by a coating only a few nanometers thick. Their results suggest that nanocoating is a promising technique to stabilize amorphous solids.
The crystallization of small-molecule APIs plays an important role in the pharmaceutical industry. Because the properties of these crystals, that is, crystal habit, size distribution, and polymorphic form, influence many downstream processes, ranging from quality control testing to formulation; it is desirable to be able to consistently produce crystals with specific properties. To achieve this kind of control over the crystallization process, a thorough understanding of the underlying mechanisms is required. In this chapter, we looked at the thermodynamic driving force behind crystallization, and how it can be rearranged into a supersaturation driving force. Methods to estimate the degree of supersaturation were discussed. The mechanisms behind crystal nucleation and growth, and the mathematical models describing these mechanisms, were discussed and examples of how these mechanisms can be used as intervention points to control the properties of the resulting crystals were given. We ended this chapter with a look at amorphous solids, which have a natural tendency to crystallize back to a more stable, but less soluble form, and saw that immobilization of the molecules at the surface of these solids with even a single layer of the polymer was enough to stabilize the amorphous solid. With an increasing movement in the pharmaceutical industry toward streamlining manufacturing processes through control, the techniques discussed in this chapter might see even more general use in the future.
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Hunter",authors:[{id:"176855",title:"Dr.",name:"Wayne",middleName:null,surname:"Hunter",slug:"wayne-hunter",fullName:"Wayne Hunter"},{id:"177693",title:"Dr.",name:"Eduardo",middleName:null,surname:"Andrade",slug:"eduardo-andrade",fullName:"Eduardo Andrade"}]},{id:"43162",doi:"10.5772/52704",title:"Non-Viral Delivery Systems in Gene Therapy",slug:"non-viral-delivery-systems-in-gene-therapy",totalDownloads:4253,totalCrossrefCites:13,totalDimensionsCites:35,abstract:null,book:{id:"3509",slug:"gene-therapy-tools-and-potential-applications",title:"Gene Therapy",fullTitle:"Gene Therapy - Tools and Potential Applications"},signatures:"Alicia Rodríguez Gascón, Ana del Pozo-Rodríguez and María Ángeles Solinís",authors:[{id:"156696",title:"Dr.",name:"Alicia",middleName:null,surname:"Rodríguez Gascón",slug:"alicia-rodriguez-gascon",fullName:"Alicia Rodríguez Gascón"}]},{id:"30029",doi:"10.5772/34438",title:"The Legume Root Nodule: From Symbiotic Nitrogen Fixation to Senescence",slug:"the-legume-root-nodule-from-symbiotic-nitrogen-fixation-to-senescence",totalDownloads:4710,totalCrossrefCites:18,totalDimensionsCites:32,abstract:null,book:{id:"1446",slug:"senescence",title:"Senescence",fullTitle:"Senescence"},signatures:"Laurence Dupont, Geneviève Alloing, Olivier Pierre, Sarra El Msehli, Julie Hopkins, Didier Hérouart and Pierre Frendo",authors:[{id:"100143",title:"Dr.",name:"Pierre",middleName:null,surname:"Frendo",slug:"pierre-frendo",fullName:"Pierre Frendo"},{id:"100842",title:"Dr.",name:"Laurence",middleName:null,surname:"Dupont",slug:"laurence-dupont",fullName:"Laurence Dupont"},{id:"100848",title:"Dr.",name:"Geneviève",middleName:null,surname:"Alloing",slug:"genevieve-alloing",fullName:"Geneviève Alloing"},{id:"100852",title:"MSc.",name:"Sarra",middleName:null,surname:"El Msehli",slug:"sarra-el-msehli",fullName:"Sarra El Msehli"},{id:"100854",title:"MSc.",name:"Olivier",middleName:null,surname:"Pierre",slug:"olivier-pierre",fullName:"Olivier Pierre"},{id:"100859",title:"BSc.",name:"Julie",middleName:null,surname:"Hopkins",slug:"julie-hopkins",fullName:"Julie Hopkins"},{id:"100860",title:"Prof.",name:"Didier",middleName:null,surname:"Hérouart",slug:"didier-herouart",fullName:"Didier Hérouart"}]},{id:"49590",doi:"10.5772/61807",title:"Management of Insect Pest by RNAi — A New Tool for Crop Protection",slug:"management-of-insect-pest-by-rnai-a-new-tool-for-crop-protection",totalDownloads:3769,totalCrossrefCites:17,totalDimensionsCites:29,abstract:"The fast-growing human population requires the development of new agricultural technologies to meet consumers´ demand, while minimizing environmental impacts. Insect pests are one of the main causes for losses in agriculture production, and current control technologies based on pesticide application or the use of transgenic crops expressing Bacillus thuringiensis toxin proteins are facing efficacy challenges. Novel approaches to control pests are urgently necessary. RNA interference (RNAi) is a gene silencing mechanism triggered by providing double-stranded RNA (dsRNA), that when ingested into insects can lead to death or affect the viability of the target pest. Transgenic plants expressing dsRNA version of insect specific target genes are the new generation of resistant plants. However, the RNAi mechanism is not conserved among insect orders, and its elucidation is the key to develop commercial RNAi crops. In this chapter, we review the core RNAi pathway in insects and the dsRNA uptake, amplification, and spread of systemic silencing signals in some key insect species. We also highlight some of the experimental steps before developing an insect-pest-resistant “RNAi plant”. Lastly, we review some of the most recent development studies to control agricultural insect pests by RNAi transgenic plants.",book:{id:"5090",slug:"rna-interference",title:"RNA Interference",fullTitle:"RNA Interference"},signatures:"Thais Barros Rodrigues and Antonio Figueira",authors:[{id:"176770",title:"Dr.",name:"Thais B.",middleName:null,surname:"Rodrigues",slug:"thais-b.-rodrigues",fullName:"Thais B. Rodrigues"},{id:"176820",title:"Dr.",name:"Antonio",middleName:null,surname:"Figueira",slug:"antonio-figueira",fullName:"Antonio Figueira"}]}],mostDownloadedChaptersLast30Days:[{id:"50471",title:"Molecular Mechanisms of Skin Aging and Rejuvenation",slug:"molecular-mechanisms-of-skin-aging-and-rejuvenation",totalDownloads:5174,totalCrossrefCites:6,totalDimensionsCites:13,abstract:"The aging process in the skin is complex and influenced by more intrinsic and extrinsic factors than any other body organ. The effects of these two types of factors overlap for the most part. The combined effects of these two aging processes also affect dermal matrix alterations. The main clinical signs of skin aging include wrinkling and irregular pigmentation, which are influenced by a combination of intrinsic and extrinsic (e.g., UV radiation, heat, smoking, and pollutants) factors. Histologically, collagen decreases, and the dermis is replaced by abnormal elastic fibers as a cause of wrinkle formation through the loss of skin elasticity. There have been numerous studies of skin aging performed to elucidate the underlying molecular mechanisms and to develop various antiaging therapeutics and preventive strategies. We summarized the molecular mechanisms and treatments of skin aging. Mainly UV radiation induces ROS formation and DNA damage, leading to increased production of MMPs and decreased production of collagen in keratinocytes and fibroblasts, which reflect the central aspects of skin aging. Besides UV radiation exposure, extrinsic factors including tobacco smoking, exposure to environmental pollutants, infrared radiation, and heat contribute to premature skin aging. Like UV radiation, these factors cause ROS formation and increase expression of MMPs, thus accelerating skin aging by inducing extracellular matrix (ECM) degradation. Accumulated collagen fibrils inhibit the new collagen synthesis and account for the further degradation of the ECM through this positive feedback loop. Accumulating evidence for molecular mechanisms of skin aging should provide clinicians with an expanding spectrum of therapeutic targets in the treatment of skin aging.",book:{id:"5258",slug:"molecular-mechanisms-of-the-aging-process-and-rejuvenation",title:"Molecular Mechanisms of the Aging Process and Rejuvenation",fullTitle:"Molecular Mechanisms of the Aging Process and Rejuvenation"},signatures:"Miri Kim and Hyun Jeong Park",authors:[{id:"47695",title:"Prof.",name:"Hyun Jeong",middleName:null,surname:"Park",slug:"hyun-jeong-park",fullName:"Hyun Jeong Park"},{id:"185767",title:"Prof.",name:"Miri",middleName:null,surname:"Kim",slug:"miri-kim",fullName:"Miri Kim"}]},{id:"49637",title:"RNA Interference Technology — Applications and Limitations",slug:"rna-interference-technology-applications-and-limitations",totalDownloads:4097,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"RNA interference (RNAi), an evolutionarily conserved mechanism triggered by double-stranded RNA (dsRNA), causes gene silencing in a sequence-specific manner. RNAi evolved naturally to mediate protection from both endogenous and exogenous pathogenic nucleic acids and to modulate gene expression. Multiple technological advancements and precision in gene targeting have allowed a plethora of potential applications, ranging from targeting infections in crop plants to improving health in human patients, which have been reviewed in this chapter.",book:{id:"5090",slug:"rna-interference",title:"RNA Interference",fullTitle:"RNA Interference"},signatures:"Devi Singh, Sarika Chaudhary, Rajendra Kumar, Preeti Sirohi,\nKamiya Mehla, Anil Sirohi, Shashi Kumar, Pooran Chand and Pankaj\nKumar Singh",authors:[{id:"176625",title:"Prof.",name:"Devi",middleName:null,surname:"Singh",slug:"devi-singh",fullName:"Devi Singh"},{id:"176744",title:"Ms.",name:"Preeti",middleName:null,surname:"Sirohi",slug:"preeti-sirohi",fullName:"Preeti Sirohi"},{id:"176745",title:"Dr.",name:"Rajendra",middleName:null,surname:"Kumar",slug:"rajendra-kumar",fullName:"Rajendra Kumar"},{id:"176746",title:"Mrs.",name:"Sarika",middleName:null,surname:"Chaudhary",slug:"sarika-chaudhary",fullName:"Sarika Chaudhary"},{id:"176747",title:"Dr.",name:"Kamiya",middleName:null,surname:"Mehla",slug:"kamiya-mehla",fullName:"Kamiya Mehla"},{id:"176748",title:"Dr.",name:"Pankaj Kumar",middleName:null,surname:"Singh",slug:"pankaj-kumar-singh",fullName:"Pankaj Kumar Singh"},{id:"176749",title:"Dr.",name:"Shashi",middleName:null,surname:"Kumar",slug:"shashi-kumar",fullName:"Shashi Kumar"},{id:"176809",title:"Dr.",name:"Pooran",middleName:null,surname:"Chand",slug:"pooran-chand",fullName:"Pooran Chand"}]},{id:"43280",title:"Gene Therapy for Diabetic Retinopathy – Targeting the Renin-Angiotensin System",slug:"gene-therapy-for-diabetic-retinopathy-targeting-the-renin-angiotensin-system",totalDownloads:2458,totalCrossrefCites:2,totalDimensionsCites:2,abstract:null,book:{id:"3509",slug:"gene-therapy-tools-and-potential-applications",title:"Gene Therapy",fullTitle:"Gene Therapy - Tools and Potential Applications"},signatures:"Qiuhong Li, Amrisha Verma, Ping Zhu, Bo Lei, Yiguo Qiu, Takahiko Nakagawa, Mohan K Raizada and William W Hauswirth",authors:[{id:"155578",title:"Dr.",name:"Qiuhong",middleName:null,surname:"Li",slug:"qiuhong-li",fullName:"Qiuhong Li"}]},{id:"49416",title:"Microinjection-Based RNA Interference Method in the Water Flea, Daphnia pulex and Daphnia magna",slug:"microinjection-based-rna-interference-method-in-the-water-flea-daphnia-pulex-and-daphnia-magna",totalDownloads:2186,totalCrossrefCites:5,totalDimensionsCites:7,abstract:"It is well known that most daphnid species have several attractive life history characteristics such as cyclical parthenogenesis, environmental sex determination, and predator-induced defense formation. Recent advances in high-throughput omics technologies make it easy to obtain a huge number of potential candidate factors involved in environmental stimuli-triggered phenotypic alterations. Furthermore, our group has developed a microinjection system to introduce foreign materials such as nucleotides and chemicals into the early-stage (one-cell stage) egg of Daphnia pulex and Daphnia magna. Consequently, we established a microinjection-based RNAi system that allows arbitrary gene functions to be investigated. However, this microinjection system does not seem to have pervaded in the daphnid research community due to its low throughput and high level of skills required. In this chapter, we review the microinjection method and its RNAi system in water fleas, D. pulex and D. magna, providing some technical tips and making challenging proposals for the development of novel high-throughput RNAi methods. Finally, we provide an overview of recently developed gene functional analysis methods such as overexpression and genome-editing systems.",book:{id:"5090",slug:"rna-interference",title:"RNA Interference",fullTitle:"RNA Interference"},signatures:"Kenji Toyota, Shinichi Miyagawa, Yukiko Ogino and Taisen Iguchi",authors:[{id:"92826",title:"Dr.",name:"Taisen",middleName:null,surname:"Iguchi",slug:"taisen-iguchi",fullName:"Taisen Iguchi"},{id:"176835",title:"Dr.",name:"Kenji",middleName:null,surname:"Toyota",slug:"kenji-toyota",fullName:"Kenji Toyota"}]},{id:"55603",title:"RNA‐seq: Applications and Best Practices",slug:"rna-seq-applications-and-best-practices",totalDownloads:3743,totalCrossrefCites:7,totalDimensionsCites:8,abstract:"RNA‐sequencing (RNA‐seq) is the state‐of‐the‐art technique for transcriptome analysis that takes advantage of high‐throughput next‐generation sequencing. Although being a powerful approach, RNA‐seq imposes major challenges throughout its steps with numerous caveats. There are currently many experimental options available, and a complete comprehension of each step is critical to make right decisions and avoid getting into inconclusive results. A complete workflow consists of: (1) experimental design; (2) sample and library preparation; (3) sequencing; and (4) data analysis. RNA‐seq enables a wide range of applications such as the discovery of novel genes, gene/transcript quantification, and differential expression and functional analysis. This chapter will encompass the main aspects from sample preparation to downstream data analysis. It will be discussed how to obtain high‐quality samples, replicates amount, library preparation, sequencing platforms and coverage, focusing on best recommended practices based on specialized literature. Basic techniques and well‐known algorithms are presented and discussed, guiding both beginners and experienced users in the implementation of reliable experiments.",book:{id:"5944",slug:"applications-of-rna-seq-and-omics-strategies-from-microorganisms-to-human-health",title:"Applications of RNA-Seq and Omics Strategies",fullTitle:"Applications of RNA-Seq and Omics Strategies - From Microorganisms to Human Health"},signatures:"Michele Araújo Pereira, Eddie Luidy Imada and Rafael Lucas Muniz\nGuedes",authors:[{id:"202103",title:"Ph.D. Student",name:"Michele",middleName:"Araújo",surname:"Pereira",slug:"michele-pereira",fullName:"Michele Pereira"},{id:"202456",title:"MSc.",name:"Eddie Luidy",middleName:null,surname:"Imada",slug:"eddie-luidy-imada",fullName:"Eddie Luidy Imada"},{id:"202460",title:"Dr.",name:"Rafael",middleName:null,surname:"Guedes",slug:"rafael-guedes",fullName:"Rafael Guedes"}]}],onlineFirstChaptersFilter:{topicId:"419",limit:6,offset:0},onlineFirstChaptersCollection:[],onlineFirstChaptersTotal:0},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:8,limit:8,total:0},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:89,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:104,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:31,numberOfPublishedChapters:314,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:11,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:141,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:129,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:113,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:105,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:18,numberOfOpenTopics:2,numberOfUpcomingTopics:1,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:5,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:14,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}},{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. The whole process of submitting an article and editing of the submitted article goes extremely smooth and fast, the number of reads and downloads of chapters is high, and the contributions are also frequently cited.",author:{id:"55578",name:"Antonio",surname:"Jurado-Navas",institutionString:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRisIQAS/Profile_Picture_1626166543950",slug:"antonio-jurado-navas",institution:{id:"720",name:"University of Malaga",country:{id:null,name:"Spain"}}}}]},series:{item:{id:"14",title:"Artificial Intelligence",doi:"10.5772/intechopen.79920",issn:"2633-1403",scope:"Artificial Intelligence (AI) is a rapidly developing multidisciplinary research area that aims to solve increasingly complex problems. In today's highly integrated world, AI promises to become a robust and powerful means for obtaining solutions to previously unsolvable problems. 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Prior to his appointment at Stellenbosch University, he has been at the University of Pretoria, Department of Computer Science (1998-2018), where he was appointed as South Africa Research Chair in Artifical Intelligence (2007-2018), the head of the Department of Computer Science (2008-2017), and Director of the Institute for Big Data and Data Science (2017-2018). 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He is a full professor of signal processing and pattern recognition and is head of the Signals and Communications Department at ULPGC, teaching from 2001 on subjects on signal processing and learning theory. His research lines are biometrics, biomedical signals and images, data mining, classification system, signal and image processing, machine learning, and environmental intelligence. He has researched in 52 international and Spanish research projects, some of them as head researcher. He is co-author of 4 books, co-editor of 27 proceedings books, guest editor for 8 JCR-ISI international journals, and up to 24 book chapters. He has over 450 papers published in international journals and conferences (81 of them indexed on JCR – ISI - Web of Science). He has published seven patents in the Spanish Patent and Trademark Office. He has been a supervisor on 8 Ph.D. theses (11 more are under supervision), and 130 master theses. He is the founder of The IEEE IWOBI conference series and the president of its Steering Committee, as well as the founder of both the InnoEducaTIC and APPIS conference series. He is an evaluator of project proposals for the European Union (H2020), Medical Research Council (MRC, UK), Spanish Government (ANECA, Spain), Research National Agency (ANR, France), DAAD (Germany), Argentinian Government, and the Colombian Institutions. He has been a reviewer in different indexed international journals (<70) and conferences (<250) since 2001. He has been a member of the IASTED Technical Committee on Image Processing from 2007 and a member of the IASTED Technical Committee on Artificial Intelligence and Expert Systems from 2011. \n\nHe has held the general chair position for the following: ACM-APPIS (2020, 2021), IEEE-IWOBI (2019, 2020 and 2020), A PPIS (2018, 2019), IEEE-IWOBI (2014, 2015, 2017, 2018), InnoEducaTIC (2014, 2017), IEEE-INES (2013), NoLISP (2011), JRBP (2012), and IEEE-ICCST (2005)\n\nHe is an associate editor of the Computational Intelligence and Neuroscience Journal (Hindawi – Q2 JCR-ISI). He was vice dean from 2004 to 2010 in the Higher Technical School of Telecommunication Engineers at ULPGC and the vice dean of Graduate and Postgraduate Studies from March 2013 to November 2017. He won the “Catedra Telefonica” Awards in Modality of Knowledge Transfer, 2017, 2018, and 2019 editions, and awards in Modality of COVID Research in 2020.\n\nPublic References:\nResearcher ID http://www.researcherid.com/rid/N-5967-2014\nORCID https://orcid.org/0000-0002-4621-2768 \nScopus Author ID https://www.scopus.com/authid/detail.uri?authorId=6602376272\nScholar Google https://scholar.google.es/citations?user=G1ks9nIAAAAJ&hl=en \nResearchGate https://www.researchgate.net/profile/Carlos_Travieso",institutionString:null,institution:{name:"University of Las Palmas de Gran Canaria",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},{id:"23",title:"Computational Neuroscience",coverUrl:"https://cdn.intechopen.com/series_topics/covers/23.jpg",isOpenForSubmission:!0,editor:{id:"14004",title:"Dr.",name:"Magnus",middleName:null,surname:"Johnsson",slug:"magnus-johnsson",fullName:"Magnus Johnsson",profilePictureURL:"https://mts.intechopen.com/storage/users/14004/images/system/14004.png",biography:"Dr Magnus Johnsson is a cross-disciplinary scientist, lecturer, scientific editor and AI/machine learning consultant from Sweden. \n\nHe is currently at Malmö University in Sweden, but also held positions at Lund University in Sweden and at Moscow Engineering Physics Institute. \nHe holds editorial positions at several international scientific journals and has served as a scientific editor for books and special journal issues. \nHis research interests are wide and include, but are not limited to, autonomous systems, computer modeling, artificial neural networks, artificial intelligence, cognitive neuroscience, cognitive robotics, cognitive architectures, cognitive aids and the philosophy of mind. \n\nDr. Johnsson has experience from working in the industry and he has a keen interest in the application of neural networks and artificial intelligence to fields like industry, finance, and medicine. \n\nWeb page: www.magnusjohnsson.se",institutionString:null,institution:{name:"Malmö University",institutionURL:null,country:{name:"Sweden"}}},editorTwo:null,editorThree:null},{id:"24",title:"Computer Vision",coverUrl:"https://cdn.intechopen.com/series_topics/covers/24.jpg",isOpenForSubmission:!0,editor:{id:"294154",title:"Prof.",name:"George",middleName:null,surname:"Papakostas",slug:"george-papakostas",fullName:"George Papakostas",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002hYaGbQAK/Profile_Picture_1624519712088",biography:"George A. 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He has (co)authored more than 150 publications in indexed journals, international conferences and book chapters, 1 book (in Greek), 3 edited books, and 5 journal special issues. His publications have more than 2100 citations with h-index 27 (GoogleScholar). His research interests include computer/machine vision, machine learning, pattern recognition, computational intelligence. \nDr. Papakostas served as a reviewer in numerous journals, as a program\ncommittee member in international conferences and he is a member of the IAENG, MIR Labs, EUCogIII, INSTICC and the Technical Chamber of Greece (TEE).",institutionString:null,institution:{name:"International Hellenic University",institutionURL:null,country:{name:"Greece"}}},editorTwo:null,editorThree:null},{id:"25",title:"Evolutionary Computation",coverUrl:"https://cdn.intechopen.com/series_topics/covers/25.jpg",isOpenForSubmission:!0,editor:{id:"136112",title:"Dr.",name:"Sebastian",middleName:null,surname:"Ventura Soto",slug:"sebastian-ventura-soto",fullName:"Sebastian Ventura Soto",profilePictureURL:"https://mts.intechopen.com/storage/users/136112/images/system/136112.png",biography:"Sebastian Ventura is a Spanish researcher, a full professor with the Department of Computer Science and Numerical Analysis, University of Córdoba. Dr Ventura also holds the positions of Affiliated Professor at Virginia Commonwealth University (Richmond, USA) and Distinguished Adjunct Professor at King Abdulaziz University (Jeddah, Saudi Arabia). Additionally, he is deputy director of the Andalusian Research Institute in Data Science and Computational Intelligence (DaSCI) and heads the Knowledge Discovery and Intelligent Systems Research Laboratory. He has published more than ten books and over 300 articles in journals and scientific conferences. Currently, his work has received over 18,000 citations according to Google Scholar, including more than 2200 citations in 2020. In the last five years, he has published more than 60 papers in international journals indexed in the JCR (around 70% of them belonging to first quartile journals) and he has edited some Springer books “Supervised Descriptive Pattern Mining” (2018), “Multiple Instance Learning - Foundations and Algorithms” (2016), and “Pattern Mining with Evolutionary Algorithms” (2016). He has also been involved in more than 20 research projects supported by the Spanish and Andalusian governments and the European Union. He currently belongs to the editorial board of PeerJ Computer Science, Information Fusion and Engineering Applications of Artificial Intelligence journals, being also associate editor of Applied Computational Intelligence and Soft Computing and IEEE Transactions on Cybernetics. Finally, he is editor-in-chief of Progress in Artificial Intelligence. He is a Senior Member of the IEEE Computer, the IEEE Computational Intelligence, and the IEEE Systems, Man, and Cybernetics Societies, and the Association of Computing Machinery (ACM). Finally, his main research interests include data science, computational intelligence, and their applications.",institutionString:null,institution:{name:"University of Córdoba",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},{id:"26",title:"Machine Learning and Data Mining",coverUrl:"https://cdn.intechopen.com/series_topics/covers/26.jpg",isOpenForSubmission:!0,editor:{id:"24555",title:"Dr.",name:"Marco Antonio",middleName:null,surname:"Aceves Fernandez",slug:"marco-antonio-aceves-fernandez",fullName:"Marco Antonio Aceves Fernandez",profilePictureURL:"https://mts.intechopen.com/storage/users/24555/images/system/24555.jpg",biography:"Dr. Marco Antonio Aceves Fernandez obtained his B.Sc. (Eng.) in Telematics from the Universidad de Colima, Mexico. 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