Characteristics of tuberculous ascites.
\\n\\n
IntechOpen was founded by scientists, for scientists, in order to make book publishing accessible around the globe. Over the last two decades, this has driven Open Access (OA) book publishing whilst levelling the playing field for global academics. Through our innovative publishing model and the support of the research community, we have now published over 5,700 Open Access books and are visited online by over three million academics every month. These researchers are increasingly working in broad technology-based subjects, driving multidisciplinary academic endeavours into human health, environment, and technology.
\\n\\nBy listening to our community, and in order to serve these rapidly growing areas which lie at the core of IntechOpen's expertise, we are launching a portfolio of Open Science journals:
\\n\\nAll three journals will publish under an Open Access model and embrace Open Science policies to help support the changing needs of academics in these fast-moving research areas. There will be direct links to preprint servers and data repositories, allowing full reproducibility and rapid dissemination of published papers to help accelerate the pace of research. Each journal has renowned Editors in Chief who will work alongside a global Editorial Board, delivering robust single-blind peer review. Supported by our internal editorial teams, this will ensure our authors will receive a quick, user-friendly, and personalised publishing experience.
\\n\\n"By launching our journals portfolio we are introducing new, dedicated homes for interdisciplinary technology-focused researchers to publish their work, whilst embracing Open Science and creating a unique global home for academics to disseminate their work. We are taking a leap toward Open Science continuing and expanding our fundamental commitment to openly sharing scientific research across the world, making it available for the benefit of all." Dr. Sara Uhac, IntechOpen CEO
\\n\\n"Our aim is to promote and create better science for a better world by increasing access to information and the latest scientific developments to all scientists, innovators, entrepreneurs and students and give them the opportunity to learn, observe and contribute to knowledge creation. Open Science promotes a swifter path from research to innovation to produce new products and services." Alex Lazinica, IntechOpen founder
\\n\\nIn conclusion, Natalia Reinic Babic, Head of Journal Publishing and Open Science at IntechOpen adds:
\\n\\n“On behalf of the journal team I’d like to thank all our Editors in Chief, Editorial Boards, internal supporting teams, and our scientific community for their continuous support in making this portfolio a reality - we couldn’t have done it without you! With your support in place, we are confident these journals will become as impactful and successful as our book publishing program and bring us closer to a more open (science) future.”
\\n\\nWe invite you to visit the journals homepage and learn more about the journal’s Editorial Boards, scope and vision as all three journals are now open for submissions.
\\n\\nFeel free to share this news on social media and help us mark this memorable moment!
\\n\\n\\n"}]',published:!0,mainMedia:{caption:"",originalUrl:"/media/original/237"}},components:[{type:"htmlEditorComponent",content:'
After years of being acknowledged as the world's leading publisher of Open Access books, today, we are proud to announce we’ve successfully launched a portfolio of Open Science journals covering rapidly expanding areas of interdisciplinary research.
\n\n\n\nIntechOpen was founded by scientists, for scientists, in order to make book publishing accessible around the globe. Over the last two decades, this has driven Open Access (OA) book publishing whilst levelling the playing field for global academics. Through our innovative publishing model and the support of the research community, we have now published over 5,700 Open Access books and are visited online by over three million academics every month. These researchers are increasingly working in broad technology-based subjects, driving multidisciplinary academic endeavours into human health, environment, and technology.
\n\nBy listening to our community, and in order to serve these rapidly growing areas which lie at the core of IntechOpen's expertise, we are launching a portfolio of Open Science journals:
\n\nAll three journals will publish under an Open Access model and embrace Open Science policies to help support the changing needs of academics in these fast-moving research areas. There will be direct links to preprint servers and data repositories, allowing full reproducibility and rapid dissemination of published papers to help accelerate the pace of research. Each journal has renowned Editors in Chief who will work alongside a global Editorial Board, delivering robust single-blind peer review. Supported by our internal editorial teams, this will ensure our authors will receive a quick, user-friendly, and personalised publishing experience.
\n\n"By launching our journals portfolio we are introducing new, dedicated homes for interdisciplinary technology-focused researchers to publish their work, whilst embracing Open Science and creating a unique global home for academics to disseminate their work. We are taking a leap toward Open Science continuing and expanding our fundamental commitment to openly sharing scientific research across the world, making it available for the benefit of all." Dr. Sara Uhac, IntechOpen CEO
\n\n"Our aim is to promote and create better science for a better world by increasing access to information and the latest scientific developments to all scientists, innovators, entrepreneurs and students and give them the opportunity to learn, observe and contribute to knowledge creation. Open Science promotes a swifter path from research to innovation to produce new products and services." Alex Lazinica, IntechOpen founder
\n\nIn conclusion, Natalia Reinic Babic, Head of Journal Publishing and Open Science at IntechOpen adds:
\n\n“On behalf of the journal team I’d like to thank all our Editors in Chief, Editorial Boards, internal supporting teams, and our scientific community for their continuous support in making this portfolio a reality - we couldn’t have done it without you! With your support in place, we are confident these journals will become as impactful and successful as our book publishing program and bring us closer to a more open (science) future.”
\n\nWe invite you to visit the journals homepage and learn more about the journal’s Editorial Boards, scope and vision as all three journals are now open for submissions.
\n\nFeel free to share this news on social media and help us mark this memorable moment!
\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:"10178",leadTitle:null,fullTitle:"Environmental Emissions",title:"Environmental Emissions",subtitle:null,reviewType:"peer-reviewed",abstract:"Today, the issue of environmental emissions is more important than ever before. Air pollution with particulates, soot, carbon, aerosols, heavy metals, and so on is causing adverse effects on human health as well as the environment. This book presents new research and findings related to environmental emissions, pollution, and future sustainability. Written by experts in the field, chapters cover such topics as health effects, emission monitoring and mitigation, and emission composition and measurement.",isbn:"978-1-83968-511-8",printIsbn:"978-1-83968-510-1",pdfIsbn:"978-1-83968-512-5",doi:"10.5772/intechopen.90676",price:119,priceEur:129,priceUsd:155,slug:"environmental-emissions",numberOfPages:162,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"febf21ec717bfe20ae25a9dab9b5d438",bookSignature:"Richard Viskup",publishedDate:"January 7th 2021",coverURL:"https://cdn.intechopen.com/books/images_new/10178.jpg",numberOfDownloads:4652,numberOfWosCitations:3,numberOfCrossrefCitations:5,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:11,numberOfDimensionsCitationsByBook:0,hasAltmetrics:1,numberOfTotalCitations:19,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"December 6th 2019",dateEndSecondStepPublish:"December 27th 2019",dateEndThirdStepPublish:"February 25th 2020",dateEndFourthStepPublish:"May 15th 2020",dateEndFifthStepPublish:"July 14th 2020",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"103742",title:"Dr.",name:"Richard",middleName:null,surname:"Viskup",slug:"richard-viskup",fullName:"Richard Viskup",profilePictureURL:"https://mts.intechopen.com/storage/users/103742/images/7778_n.jpg",biography:"Richard Viskup was born in Bratislava in the Slovak Republic, formerly Czechoslovakia. He received his Master of Science, Doctor in Natural Science, and Doctor of Philosophy in Physics, Plasma Physics, and Laser Physics, respectively, from Comenius University, Bratislava. He obtained his postgraduate Master of Philosophy in Photonics from Strathclyde University, Glasgow, Scotland, and a Doctor of Engineering in Applied Physics from Johannes Kepler University, Linz, Austria.\nDr. Viskup’s research interests include physics, plasma, lasers, material science and analyses, radiation physics, analytical chemistry, spectroscopy, combustion processes, and environmental science, among others.",institutionString:"Johannes Kepler University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"5",totalChapterViews:"0",totalEditedBooks:"4",institution:{name:"Johannes Kepler University of Linz",institutionURL:null,country:{name:"Austria"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"133",title:"Environmental Pollution",slug:"environmental-pollution"}],chapters:[{id:"72552",title:"Effects of Environmental Emissions on the Respiratory System: Secrets and Consequences",doi:"10.5772/intechopen.92451",slug:"effects-of-environmental-emissions-on-the-respiratory-system-secrets-and-consequences",totalDownloads:536,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:1,abstract:"Human health has been affected adversely by air pollution as a serious environmental challenge. Ambient (outdoor) air pollution mainly resulted from human activities (e.g., fuel combustion, heat generation, industrial facilities) causes 4.2 million deaths every year. Moreover, each year, 3.8 million people die from indoor air pollution which means household exposure to smoke from fuels and dirty cook stoves. They are the risks of stroke, heart attack, lung disease, or cancer that resulted from air pollution which assaults our brain, heart, and lungs using its invisible weapons named particulate matter (PM). These inhalable particles are of a nanoscale or microscale size. Upon inhalation, the air with its components enters the human body through the respiratory system. The lungs are the responsible organs for gas exchange with blood. Inhaled particles, such as silica, organic compounds, and metallic dusts, have toxic effects on our pulmonary system. For example, the accumulation of nanoparticles in the kidneys, liver, spleen, and central nervous system through the penetration of the epithelial barriers in the lungs has been observed. The purpose of this chapter is to describe the toxic effects of air particles on the different organs in the human body and to introduce some of the adverse effects of air pollution on human health.",signatures:"Farzaneh Hajirasouliha and Dominika Zabiegaj",downloadPdfUrl:"/chapter/pdf-download/72552",previewPdfUrl:"/chapter/pdf-preview/72552",authors:[{id:"317989",title:"M.Sc.",name:"Farzaneh",surname:"Hajirasouliha",slug:"farzaneh-hajirasouliha",fullName:"Farzaneh Hajirasouliha"},{id:"320479",title:"Dr.",name:"Dominika",surname:"Zabiegaj",slug:"dominika-zabiegaj",fullName:"Dominika Zabiegaj"}],corrections:null},{id:"74104",title:"Health Effect of Biomass Fuel Smoke",doi:"10.5772/intechopen.94611",slug:"health-effect-of-biomass-fuel-smoke",totalDownloads:535,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:1,abstract:"Almost half of the world population rely on solid (biomass fuel and coal) for cooking, heating and lightning purpose. The resultant exposure to fine particulate matter from household air pollution is the seventh-largest risk factor for global burden of disease causing between 2.6 and 3.8 million premature deaths per year. The health effect ranges from cardiovascular, respiratory, neurocognitive and reproductive health effect. The most important are cardiovascular and respiratory health effects; others are the risk of burns and cataract in the eyes. Biomass fuel is any living or recently living plant and animal-based material that is burned by humans as fuels, for example, wood, dried animal dung, charcoal, grass and other agricultural residues. Biomass fuels are at the low end of the energy ladder in terms of combustion efficiency and cleanliness. Incomplete combustion of biomass contributes majorly to household air pollution and ambient air pollution. A large number of health-damaging air pollutants are produced during the incomplete combustion of biomass. These include respirable particulate matter, carbon monoxide, nitrogen oxides, formaldehyde, benzene, 1, 3 butadiene, polycyclic aromatic hydrocarbons (PAHs), and many other toxic organic compounds. In this article, health effects of biomass fuel use will be described in details highlighting the most affected systems and organs of the body.",signatures:"Olayemi Fehintola Awopeju",downloadPdfUrl:"/chapter/pdf-download/74104",previewPdfUrl:"/chapter/pdf-preview/74104",authors:[{id:"323972",title:"Dr.",name:"Olayemi Fehintola",surname:"Awopeju",slug:"olayemi-fehintola-awopeju",fullName:"Olayemi Fehintola Awopeju"}],corrections:null},{id:"72031",title:"Importance of Air Quality Networks in Controlling Exposure to Air Pollution",doi:"10.5772/intechopen.92335",slug:"importance-of-air-quality-networks-in-controlling-exposure-to-air-pollution",totalDownloads:577,totalCrossrefCites:0,totalDimensionsCites:2,hasAltmetrics:0,abstract:"An air quality monitoring network (AQMN) is a basic piece of environmental management due to that it satisfies the major role in monitoring of environment emissions, in special relevance to target air pollutants. An adequate installation would lead to support high efficiency of the network. Therefore, AQMN pre-layout should be considered as an essential factor in regarding with the location of fixed measurement stations within AQMN, as the minimum number of sampling points. Nevertheless, once AQMN has been already installed, and given that the spatial air pollutants pattern can vary along time, an assessment of the AQMN design would be addressed in order to identify the presence of potential redundant fixed monitoring stations. This approach would let to improve the AQMN performance, reduce maintenance costs of the network and consolidate the investment on those more efficient fixed stations. The chapter includes aspects relative to air pollutants measured by networks, their representativeness, limitations, importance, and the future needs. It ponders the need of re-assessment of the AQMN layout for assuring (i) a right evaluation of the human being exposure to atmospheric pollutants and controlling the environmental emissions into the atmosphere and (ii) an adequate performance of the network along time.",signatures:"David Galán Madruga",downloadPdfUrl:"/chapter/pdf-download/72031",previewPdfUrl:"/chapter/pdf-preview/72031",authors:[{id:"316730",title:"Dr.",name:"David Galan",surname:"Madruga",slug:"david-galan-madruga",fullName:"David Galan Madruga"}],corrections:null},{id:"72766",title:"Industrial Air Emission Pollution: Potential Sources and Sustainable Mitigation",doi:"10.5772/intechopen.93104",slug:"industrial-air-emission-pollution-potential-sources-and-sustainable-mitigation",totalDownloads:920,totalCrossrefCites:2,totalDimensionsCites:4,hasAltmetrics:1,abstract:"Air of cities especially in the developing parts of the world is turning into a serious environmental interest. The air pollution is because of a complex interaction of dispersion and emission of toxic pollutants from manufactories. Air pollution caused due to the introduction of dust particles, gases, and smoke into the atmosphere exceeds the air quality levels. Air pollutants are the precursor of photochemical smog and acid rain that causes the asthmatic problems leading into serious illness of lung cancer, depletes the stratospheric ozone, and contributes in global warming. In the present industrial economy era, air pollution is an unavoidable product that cannot be completely removed but stern actions can reduce it. Pollution can be reduced through collective as well as individual contributions. There are multiple sources of air pollution, which are industries, fossil fuels, agro waste, and vehicular emissions. Industrial processes upgradation, energy efficiency, agricultural waste burning control, and fuel conversion are important aspects to reducing pollutants which create the industrial air pollution. Mitigations are necessary to reduce the threat of air pollution using the various applicable technologies like CO2 sequestering, industrial energy efficiency, improving the combustion processes of the vehicular engines, and reducing the gas production from agriculture cultivations.",signatures:"Rabia Munsif, Muhammad Zubair, Ayesha Aziz and Muhammad Nadeem Zafar",downloadPdfUrl:"/chapter/pdf-download/72766",previewPdfUrl:"/chapter/pdf-preview/72766",authors:[{id:"251787",title:"Dr.",name:"Muhammad",surname:"Zubair",slug:"muhammad-zubair",fullName:"Muhammad Zubair"},{id:"318519",title:"Ms.",name:"Rabia",surname:"Munsif",slug:"rabia-munsif",fullName:"Rabia Munsif"},{id:"320637",title:"Ms.",name:"Ayesha",surname:"Aziz",slug:"ayesha-aziz",fullName:"Ayesha Aziz"},{id:"320675",title:"Dr.",name:"Muhammad Nadeem",surname:"Zafar",slug:"muhammad-nadeem-zafar",fullName:"Muhammad Nadeem Zafar"}],corrections:null},{id:"72129",title:"Methods to Reduce Mercury and Nitrogen Oxides Emissions from Coal Combustion Processes",doi:"10.5772/intechopen.92342",slug:"methods-to-reduce-mercury-and-nitrogen-oxides-emissions-from-coal-combustion-processes",totalDownloads:679,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The chapter presents the issue of reducing mercury and nitrogen oxides emissions from the flue gas of coal-fired boilers. The issue is particularly relevant due to the stricter regulations regarding exhaust gas purity. A brief review of the methods for reducing Hg and NOx emissions has been made, pointing out their pros and cons. Against this background, the results of the authors’ own research on the injection of selected oxidants into flue gases to remove both of these pollutants are presented. The injection of sodium chlorite solution into the flue gas (400 MWe lignite fired unit) upstream the wet flue gas desulphurization (WFGD) absorber contributed to the oxidation of both metallic mercury and nitric oxide and enhanced their removal efficiency. The results of tests on lignite and hard coal flue gases indicate that in order to reduce the unfavorable phenomenon of mercury re-emission from WFGD absorbers, in some cases, it is necessary to add selected chemical compounds (e.g., sulfides) to the desulfurization system. The results of field tests for flue gas from lignite (400 MWe unit) and hard coal-fired boilers (195 and 220 MWe units) confirmed the usefulness of oxidizer injection technology to reduce mercury emissions below the level required by BAT conclusions.",signatures:"Maria Jędrusik, Dariusz Łuszkiewicz and Arkadiusz Świerczok",downloadPdfUrl:"/chapter/pdf-download/72129",previewPdfUrl:"/chapter/pdf-preview/72129",authors:[{id:"317074",title:"Prof.",name:"Maria",surname:"Jedrusik",slug:"maria-jedrusik",fullName:"Maria Jedrusik"},{id:"317075",title:"Dr.",name:"Dariusz",surname:"Luszkiewicz",slug:"dariusz-luszkiewicz",fullName:"Dariusz Luszkiewicz"},{id:"317076",title:"Prof.",name:"Arkadiusz",surname:"Swierczok",slug:"arkadiusz-swierczok",fullName:"Arkadiusz Swierczok"}],corrections:null},{id:"72386",title:"Carbon Soot Polymer Nanocomposites (CSPNCs): Production, Surface Morphological, Glass Transition Temperature Phenomenon and Optical Properties",doi:"10.5772/intechopen.92389",slug:"carbon-soot-polymer-nanocomposites-cspncs-production-surface-morphological-glass-transition-temperat",totalDownloads:529,totalCrossrefCites:2,totalDimensionsCites:4,hasAltmetrics:0,abstract:"The present chapter covers the production and properties of carbon soot nanoparticles (CSNPCs) and their doped carbon soot polymer nanocomposites (CSPNCs). The first part of this chapter will provide a brief introduction of carbon soot, its morphology, production and synthesis methods. The second part will explain the investigation of carbon soot nanoparticles by flame deposition method and their properties. The third part will provide a short knowledge on polymer nanocomposites (PNCs) and their processing methods. The last part will illustrate the production of carbon soot polymer nanocomposites by solution casting method and their important properties. At the end, the chapter concludes with future scopes.",signatures:"Rakhi Tailor, Yogesh Kumar Vijay and Minal Bafna",downloadPdfUrl:"/chapter/pdf-download/72386",previewPdfUrl:"/chapter/pdf-preview/72386",authors:[{id:"316784",title:"Dr.",name:"Rakhi",surname:"Tailor",slug:"rakhi-tailor",fullName:"Rakhi Tailor"},{id:"319806",title:"Prof.",name:"Yogesh Kumar",surname:"Vijay",slug:"yogesh-kumar-vijay",fullName:"Yogesh Kumar Vijay"},{id:"319843",title:"Dr.",name:"Minal",surname:"Bafna",slug:"minal-bafna",fullName:"Minal Bafna"}],corrections:null},{id:"72246",title:"The Use of Synchronous Fluorescence Technique in Environmental Investigations of Polycyclic Aromatic Hydrocarbons in Airborne Particulate Matter from an Industrial Region in Poland",doi:"10.5772/intechopen.92402",slug:"the-use-of-synchronous-fluorescence-technique-in-environmental-investigations-of-polycyclic-aromatic",totalDownloads:490,totalCrossrefCites:1,totalDimensionsCites:1,hasAltmetrics:0,abstract:"The applicability of the fluorescence techniques to identify the polycyclic aromatic hydrocarbons (PAHs) in environmental samples is presented. The technique of synchronous fluorescence enabled the identification of the PAHs series containing 2–6 condensed rings in urban airborne particulate matter from Upper Silesia industrial region in Poland. The results obtained by synchronous and conventional fluorescence measurements have been confirmed by those from gas chromatography-mass spectrometry. As the air sample was taken in summer season, the main source of pollution by PAHs component seems to be transport – the exhaust gases from motor vehicles.",signatures:"Aniela Matuszewska and Maria Czaja",downloadPdfUrl:"/chapter/pdf-download/72246",previewPdfUrl:"/chapter/pdf-preview/72246",authors:[{id:"40700",title:"Dr.",name:"Aniela",surname:"Matuszewska",slug:"aniela-matuszewska",fullName:"Aniela Matuszewska"},{id:"40798",title:"Prof.",name:"Maria",surname:"Czaja",slug:"maria-czaja",fullName:"Maria Czaja"}],corrections:null},{id:"72658",title:"Qualitative Characterisation of Trace Elements in Diesel Particulate Matter from In-Use Diesel Engine Passenger Vehicles by Means of Laser-Induced Breakdown Spectroscopy",doi:"10.5772/intechopen.93067",slug:"qualitative-characterisation-of-trace-elements-in-diesel-particulate-matter-from-in-use-diesel-engin",totalDownloads:387,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"In this research, we applied laser-plasma spectroscopy technique for the measurement of trace chemical elements in the exhaust emissions generated from in-use diesel engine passenger vehicles. We use high resolution laser-induced breakdown spectroscopy (LIBS) technique for diagnostics of soot and particulate matter (PM). Here we analysed soot and PM, extracted from exhaust manifold part, from different passenger vehicles that are used in daily life environment. The main aim of this study is to reveal the trace chemical elements in different PM matrices. The presence of trace elements in exhaust emissions can originate from different sources: from injected fuel type and fuel additives, engine lubricants, engine combustion process, incomplete catalytic reaction, inefficiency or wear out of PM filtering devices, dysfunctions or failures of engine or vehicle or even information related to polluted intake air.",signatures:"Richard Viskup, Christoph Wolf and Werner Baumgartner",downloadPdfUrl:"/chapter/pdf-download/72658",previewPdfUrl:"/chapter/pdf-preview/72658",authors:[{id:"103742",title:"Dr.",name:"Richard",surname:"Viskup",slug:"richard-viskup",fullName:"Richard Viskup"},{id:"305127",title:"M.Sc.",name:"Christoph",surname:"Wolf",slug:"christoph-wolf",fullName:"Christoph Wolf"},{id:"310228",title:"Dr.",name:"Werner",surname:"Baumgartner",slug:"werner-baumgartner",fullName:"Werner Baumgartner"}],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},subseries:null,tags:null},relatedBooks:[{type:"book",id:"5236",title:"High Energy and Short Pulse Lasers",subtitle:null,isOpenForSubmission:!1,hash:"481d4221e58d2c90fe398be93d898f43",slug:"high-energy-and-short-pulse-lasers",bookSignature:"Richard Viskup",coverURL:"https://cdn.intechopen.com/books/images_new/5236.jpg",editedByType:"Edited by",editors:[{id:"103742",title:"Dr.",name:"Richard",surname:"Viskup",slug:"richard-viskup",fullName:"Richard Viskup"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7377",title:"Diesel and Gasoline Engines",subtitle:null,isOpenForSubmission:!1,hash:"dab9fe312a28dd603ac4b21628070d59",slug:"diesel-and-gasoline-engines",bookSignature:"Richard Viskup",coverURL:"https://cdn.intechopen.com/books/images_new/7377.jpg",editedByType:"Edited by",editors:[{id:"103742",title:"Dr.",name:"Richard",surname:"Viskup",slug:"richard-viskup",fullName:"Richard Viskup"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8813",title:"Introduction to Diesel Emissions",subtitle:null,isOpenForSubmission:!1,hash:"693a8757f50c6f257cca62961cba76c2",slug:"introduction-to-diesel-emissions",bookSignature:"Richard Viskup",coverURL:"https://cdn.intechopen.com/books/images_new/8813.jpg",editedByType:"Edited by",editors:[{id:"103742",title:"Dr.",name:"Richard",surname:"Viskup",slug:"richard-viskup",fullName:"Richard Viskup"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7547",title:"Monitoring of Marine Pollution",subtitle:null,isOpenForSubmission:!1,hash:"4700c71016d4ab73a99b22cee68da2fe",slug:"monitoring-of-marine-pollution",bookSignature:"Houma Bachari Fouzia",coverURL:"https://cdn.intechopen.com/books/images_new/7547.jpg",editedByType:"Edited by",editors:[{id:"95997",title:"Dr.",name:"Houma",surname:"Bachari Fouzia",slug:"houma-bachari-fouzia",fullName:"Houma Bachari Fouzia"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8863",title:"Hydrocarbon Pollution and its Effect on the Environment",subtitle:null,isOpenForSubmission:!1,hash:"25243b6684e6a441a6bf1f854d49f9e8",slug:"hydrocarbon-pollution-and-its-effect-on-the-environment",bookSignature:"Muharrem Ince and Olcay Kaplan Ince",coverURL:"https://cdn.intechopen.com/books/images_new/8863.jpg",editedByType:"Edited by",editors:[{id:"258431",title:"Prof.",name:"Muharrem",surname:"Ince",slug:"muharrem-ince",fullName:"Muharrem Ince"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7224",title:"Persistent Organic Pollutants",subtitle:null,isOpenForSubmission:!1,hash:"06c3095a17bf790c56c71013cc5e3ad6",slug:"persistent-organic-pollutants",bookSignature:"Stephen Kudom Donyinah",coverURL:"https://cdn.intechopen.com/books/images_new/7224.jpg",editedByType:"Edited by",editors:[{id:"26196",title:"Dr.",name:"Stephen Kudom",surname:"Donyinah",slug:"stephen-kudom-donyinah",fullName:"Stephen Kudom Donyinah"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"6644",title:"Emerging Pollutants",subtitle:"Some Strategies for the Quality Preservation of Our Environment",isOpenForSubmission:!1,hash:"9e03aeca8b09510ef11fcf3621a2a996",slug:"emerging-pollutants-some-strategies-for-the-quality-preservation-of-our-environment",bookSignature:"Sonia Soloneski and Marcelo L. 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Tuberculosis (TB) is a dangerous infection affecting about one third of the world population despite the availability of affordable and effective chemotherapy, remaining one of the major causes of death from a single infectious agent worldwide. The most affected organ is the lung. It is preventable through Bacillus of Calmette and Guérin (BCG) vaccination and curable with antituberculous drugs.
Tuberculosis is a serious and highly contagious bacterial infection which in humans is usually caused by bacteria called
The absolute number of incident cases has been decreasing since the early 2000s. The lowest incidence rate is found in high-income countries including the United States of America, Canada, New Zealand, Western Europe, and Australia. The largest number of incident cases is found in low- and middle-income countries. In 2015, 61% of the new cases occurred in Asia, followed by 26% new cases in Africa (Figure 1) [2].
Worldwide incidence of TB infections.
The HIV infection is the prevalent risk factor for the development of TB because HIV alters the pathogenesis of TB by producing a progressive decline in cell-mediated immunity and raises the chances of extrapulmonary involvement [3, 4].
Tuberculosis is a disease which typically involves the respiratory system, being characterized by the growth of tubercles in tissues, but it can affect any other organ, in which case it’s called extrapulmonary tuberculosis (EPTB) and usually results from hematogenous dissemination, being particularly present in immunocompromised patients. In some cases the infection directly extends from an adjacent organ. The most common sites of extrapulmonary tuberculosis are the lymph nodes, abdomen, bones and joints, pleura, spinal cord, and brain [5, 6].
Abdominal tuberculosis is a common form of extrapulmonary tuberculosis. The infection involving
Peritoneal tuberculosis is an uncommon site of extrapulmonary infection caused by
Infection occurs frequently following reactivation of latent tuberculous in the peritoneum from hematogenous spread from a primary lung focus. It can also occur via hematogenous spread from active pulmonary or miliary TB. Not so often, the organisms enter the peritoneal cavity transmurally, from an infected small intestine or contiguously from tuberculous salpingitis. Over time, the visceral and parietal peritoneum becomes studded with tubercles [7–9].
Ascites is defined as an abnormal accumulation of fluid in the peritoneal cavity, the presence of serous fluid between the visceral and parietal peritoneum. The word ascites is derived from the ancient Greek word “
Tuberculous ascites, one of the clinical signs of abdominal TB, implies accumulation of fluid in the abdomen, a swollen abdomen, and slightly raised tubercles of 1–2 mm all over the peritoneum. In EPTB, ascites develops secondary to “exudation” of proteinaceous fluid from the tubercles, similar to the mechanism leading to ascites in patients with peritoneal carcinomatosis, and it’s often misdiagnosed in elderly patients. Most patients with tuberculous peritonitis have ascites at the time of diagnosis, while the rest present the advanced phase, the dry or fibroadhesive form of the disease [11, 12].
Tuberculous peritonitis is a subacute disease, and its symptoms evolve over a period of several weeks or months.
The insidious onset of this condition and the fact that the diagnosis is rarely suspected explains why patients have symptoms for more than 4 months before the diagnosis is established. Tuberculous peritonitis should be considered in any patient presenting with several weeks of abdominal pain, fever, and weight loss. Systemic and constitutional manifestations are common. Symptoms may be mild, with fatigue, abdominal pain, and tenderness, or severe enough to mimic acute abdomen [13]. Other clinical manifestations could be:
Constipation/diarrhea
Nocturnal hyperhidrosis
Low-grade fever
Anorexia
Malaise
The clinical presentation of TB ascites is challenging, since it is nonspecific and can be confused with a plethora of other infectious or noninfectious diseases, leading to a delayed diagnosis and treatment which are major factors that contribute to the high mortality of TB.
Another situation that contributes to a delayed diagnosis is the presence of multiple comorbidities such as HIV/AIDS, cirrhosis, uremia, or other chronic conditions. Additional illness results in atypical presentation of TB ascites which render the symptoms more difficult to identify and distinguish. Moreover, in elderly patients, clinical manifestation is minimal with abdominal discomfort, constipation, or fatigue, symptoms that most people tend to ignore as minor or non-perilous (Figure 2) [14, 15].
Factors associated with delayed diagnosis.
Due to the fact that the nature of this disease is insidious, the diagnosis represents a challenge for clinicians. With the ever-increasing demographic shifts, more cases are now detected in areas where TB was a rarity until present. Unless a high degree of suspicion is maintained, the diagnosis can easily be missed or delayed [12].
Diagnostic techniques and procedures include:
Clinical observation
Imaging techniques: ultrasound, CT/MRI
Laboratory tests
Ascites fluid microbiologic and biochemical analysis
Generally, diagnosis is based on clinical suspicion, imaging of tuberculous infected zone, and detection of
Nucleic acid amplification tests (NAATs) are molecular diagnostic methods based on amplification of mycobacterial nucleic acid. These are rapid methods that provide results within a day, and they are more specific and sensitive than Acid-Fast Bacillus Smear (AFB) smear. Albeit NAATs were originally designed for respiratory specimens, they can also be used on specimens from other TB sites like ascitic fluid samples, but this technique is still under evaluation.
Ascites of TB peritonitis obtained through ultrasound-guided paracentesis is an exudative type, and macroscopically its appearance is chylous and cloudy or turbid. Biochemically, the serum-ascites albumin gradient (SAAG) is now considered a more sensitive and specific measure than the ascitic total protein concentration which has been used for many years, in differentiating the ascites due to portal hypertension from ascites due to other pathophysiological mechanisms, such as tuberculous ascites which has a SAAG <1.1 g/dL and total proteins >3–4 g% [18, 19]. Combining LDH with total protein analysis has been explored for ascitic fluid. The cutoff values for three parameters in the ascitic fluid for differentiation between hepatic and non-hepatic ascites are as follows: LDH of 400 Sigma units, fluid/serum total protein (TP) ratio of 0.5, and fluid/serum LDH ratio of 0.6. The presence of any two of these three findings is usually associated with TB; the absence of all three indicates a hepatic cause [20].
Glucose concentration in the ascitic fluid, under normal conditions, is similar to that in the serum. Ascitic glucose concentration decreases due to consumption by bacteria, white blood cells, or cancer cells in the fluid in TB peritonitis. Ascitic glucose concentration is lower than normal in TB ascites, which makes it an indicator in differentiating tuberculosis from other diseases, such as cirrhosis. The ascitic/blood glucose ratio is a useful test in differentiating TB peritonitis from other causes of ascites [18].
Ascitic fluid adenosine deaminase activity (ADA) is considered a more sensitive and specific method used for early diagnosis of TB ascites. Even if the full physiological role of ADA is not yet completely understood, it is known that ADA values are notably higher (>40 U/L) in patients with TB ascites [21–23].
Non-biochemical tests of ascitic fluid, including cell counts, bacterial culture, and polymerase chain reaction (PCR), have an important role in diagnosing the cause of ascites.
The total cell count in tuberculous ascites is 150–4000/μL, and the cytologic examination shows over 70–80% lymphocytes and more than 250 leucocytes/mm3 (Table 1).
Type | Exudative |
---|---|
Appearance | Chylous and cloudy/turbid |
Total cell count | 150–4000/μL |
Leucocytes | >250/mm3 |
Lymphocytes | >70–80% |
Total proteins | >3–4 g% |
ADA | >40 U/L |
LDH | >400 SU |
Glucose | <6 mg/dL |
SAAG | <1.1 g/dL |
Characteristics of tuberculous ascites.
The sensitivity of direct microscopic smear detection of acid-fast bacilli in the ascitic fluid (0–6%) and ascitic fluid mycobacterial culture (20–35%) is low, and because of the delay in obtaining the results of mycobacterial cultures of ascitic fluid, the mortality is high, and the value of these tests in the differential diagnosis of ascites is limited.
Recently, a new approach to the fast diagnosis of bacterial infections emerged, including tuberculosis. The advanced molecular techniques provided a new method represented by PCR which can detect minimal amounts of bacterial DNA and improves the rates of bacterial identification from 4 to 6 weeks for microbiological cultures to 24 hours. In diagnosing TB effusions, PCR appears to be an ideal tool, with 94% sensitivity and 88% specificity, becoming a rapid and reliable method for identification of infectious ascites [24].
The tuberculin skin testing (TST or purified protein derivative (PPD) skin test) is controversial, despite the high specificity of this test which is between 95 and 99%. Skin testing is currently used only for detection of latent infection because of its low sensitivity and low positive predictive value. At the moment, there are no recommendations for using this test to diagnose active disease like tuberculous peritonitis. At best, tuberculin skin testing can only offer auxiliary information. Several studies have reported positivity rates ranging between 24 and 100% with no significant difference between high and low endemicity areas. Currently, the recommendations about the cut point for the induration differ depending on the risk scale. For patients at low risk, the cut point is at 15 mm; in cases of moderate risk, the cut point is 10 mm, and for those at high risk, the cut point is 5 mm. Generally, about 50% of the patients would have false-negative reactions to this test, suggesting the fact that it has many potential sources of error. In conclusion, anergy testing may yield confusing information and is no longer recommended for diagnosis [25].
A great scientific advance has been the development of an IFN-c-based test with an 89% sensitivity, which is a quantitative in vitro assay evaluating the cell-mediated immune response to
In the past few years, the tuberculin skin test has been replaced by T-cell-based interferon-gamma release assay (IGRA) which is more sensitive and more specific. IGRA is an in vitro test used in all circumstances in which the TST is currently used, including evaluation of immigrants, surveillance programs, or contact investigations [26]. There are three commercially available IFN-γ tests: QuantiFERON-TB Gold assay (QFN-Gold), QuantiFERON-TB Gold
Imaging techniques used for detection of TB ascites are ultrasound and computed tomography. These methods also increase the accuracy of several procedures like paracentesis or peritoneal biopsies, providing a safer and affordable replacement to diagnostic laparoscopy [30].
Ultrasound is the most sensitive and reliable method of detecting ascites, guiding paracentesis and monitoring the effects of therapy. It can detect even small volumes of fluid (as little as 100 ml of fluid could be detected). Ascites is usually seen as an anechoic space. In TB ascites, particulate matter within the ascitic fluid or fine, mobile strands, representing echogenic debris, can be detected. Less commonly, the ultrasound can reveal calcifications in the walls of encysted ascitic fluid [12].
On computed tomography the ascitic fluid has high attenuation values, between 20 and 45 HU, and the peritoneum is symmetrically thickened and nodular. Frequently, an early sign of abdominal TB is a thickened mesentery (>15 mm) with mesenteric lymph nodes.
Many studies concluded that ultrasonography and computed tomography are complementary to each other in detecting TB ascites, as they provide different details. CT focuses on the peritoneum and omental and mesentery involvement, and the ultrasound shows fine, mobile septations (Figure 3) [31].
Circumferential parietal thickness, with an edematous appearance of the appendix, cecum, ascending colon, and up to the hepatic flexura, associating stripe thickening of adjacent fat. Multiple adenopathies containing calcifications and central necrosis. Fluid accumulation in the peritoneal cavity associating symmetrical, iodophilic thickening of parietal peritoneum. Osteolytic areas with an adjacent osteosclerotic reaction in L2, L3, and L4 vertebrae. Parafluid accumulation in L2–L3 and L3–L4 intervertebral spaces that reaches the anterior epidural space and the roots of L2 and L3 nerves, extending to L3–L4 adjacent smooth tissues, with no visible border toward the right psoas muscle.
However, the only certain way to diagnose TB ascites is the histological examination. Various methods that include excision, laparoscopy, needle biopsy and ultrasound-guided biopsies, endoscopy, computed tomography (CT), or endoscopic ultrasound are used to establish the diagnosis. The presence of granulomas is typical for TB ascites [32]. The relative sensitivities of different procedures and the potential therapeutic benefits should be considered in making the choice of diagnostic approach. In superficial TB lymphadenitis, fine needle aspiration (FNA) biopsy of affected lymph nodes is the gold-standard diagnostic technique. Excision biopsy has the highest sensitivity, whereas FNA is less invasive and may be useful. If FNA examination results are doubtful, excision biopsy may be needed. Laparoscopy with target peritoneal biopsy is the current first-line investigation in the diagnosis of peritoneal TB [33, 34]. Several studies revealed a diagnostic accuracy of 84–96% for TB peritonitis.
Generally, tissue biopsy is superior to fluid aspiration in providing positive culture results. The diagnosis is more accurate when the biopsy results and polymerase chain reaction assays are combined with culture results [35].
The main differential diagnosis is peritoneal carcinomatosis, which can be difficult to distinguish, especially in older patients. The nodules of carcinomatosis are larger, usually more than 3 mm, more vascular, and more irregular than the tuberculous ones which rarely surpass 1–2 mm. Carcinomatosis is seen as an irregular peritoneal thickening with nodular implants, while TB peritonitis is suggested by the presence of a smooth peritoneum with symmetrical thickening, ascites, and enlarged lymph nodes of low attenuation [31].
Other less likely considerations include:
Ascites in liver diseases: the liver is enlarged and irregular; proteins are lower than 4 g%.
Nephrotic syndrome: ascites is less marked; proteins are lower than 4 g%.
Nutritional edema (hypoproteinemia) has many other signs of protein deficiency; proteins are also lower than 4 g%.
Starch peritonitis, sarcoidosis, and Crohn’s disease may resemble the laparoscopic features of TB peritonitis, but the presence of caseating granuloma establishes the diagnosis (Table 2) [36, 37].
The most important differential diagnosis | Less likely considerations |
---|---|
Ascites in liver disease | |
Nephrotic syndrome | |
Peritoneal carcinomatosis | Hypoproteinemia |
Sarcoidosis | |
Starch peritonitis | |
Crohn’s disease |
Differential diagnosis.
There are a few signs that additionally suggest the diagnosis of TB peritonitis: normal serum levels of CA 19–9 and carcinoembryonic antigen (CEA), elevated serum levels of CA 125, fever, and lymphocyte-predominant benign ascites, but only biopsies yield the final diagnosis [38, 39].
Treatment is initiated not only in patients with confirmed diagnosis but also in patients with strong suspicion of TB, because a delay in treatment initiation can lead to significant mortality. TB treatment initiation includes also individuals with ascites associated with fever, weight loss, imaging signs of TB, personal history of TB, or contact with a tuberculosis case.
Despite the fact that most guidelines on the treatment of tuberculosis suggest that 6 months of treatment is sufficient for extrapulmonary tuberculosis (except for the case of bone tuberculosis or tuberculous meningitis), most physicians treating peritoneal tuberculosis use antituberculous therapy for 9 to 12 months [40, 41].
Drug treatment is the most important modality and follows standard regimens and principles. There are currently five drugs that are considered first-line medications: isoniazid, rifampicin, pyrazinamide, streptomycin, and ethambutol. Second-line drugs are only used in case of resistance to first-line therapy (extensively drug-resistant tuberculosis or multidrug-resistant tuberculosis), and they are represented by aminoglycosides, fluoroquinolones, polypeptides, cycloserine, thioamides, and terizidone. There is also a third-line therapy with uncertain or unproven efficacy including rifabutin, macrolides, linezolid, thioacetazone, thioridazine, arginine, bedaquiline, and vitamin D [42]. Drug-resistant disease varies substantially in different areas of the world and may occur in cases of poor patient management, nonadherence to prescribed regimen, or as a result of malabsorption of the antituberculous drugs.
The treatment of TB peritonitis in patients with HIV is usually the same, but because HIV-infected patients are often taking multiple drugs, some of which may interact with antituberculous ones; it is strongly recommended to consult the experts in HIV-related TB.
The “complete response” to antituberculous treatment means complete resolution of symptoms and ascites within 6 months; in most cases, laboratory tests return to normal values within 3 months. Persistence of ascites means “no response” [43, 44].
Peritoneal tuberculosis is still common in areas of the world where TB is prevalent and its incidence ratio is likely to increase as a consequence of population migrations.
Ascites can be a complication of an aggregate of diseases, which carries an unfavorable prognosis that depends on the causes, the moment of diagnosis, and the start of the treatment.
Establishing the diagnosis of TB ascites requires a high index of suspicion because of its insidious development. In any patient with several weeks of abdominal pain, weight loss, fever, and lymphocytic dominant ascites with SAAG < 1.1 g/L, as well as in patients with ascites belonging to special population groups, such as indigenous or older people, or patients with ascites as the primary symptom, ascitic TB peritonitis should be considered in differential diagnosis. This syndrome behaves clinically like many abdominal diseases that are often ignored, leading to a significant impact on morbidity and mortality due to a delayed diagnosis and treatment. Older laboratory tests lack sensitivity and specificity in establishing the diagnosis. Histological examination, considered the gold-standard diagnosis method, is an invasive procedure with high risk of complications. More accurate methods such as molecular tests based on mycobacterial nucleic acid amplification tests (NAATs), PCR techniques used to detect bacterial DNA, or interferon-gamma release assays (IGRA) and biochemical methods such as the serum-ascites albumin gradient (SAAG), ratio between LDH in ascites fluid/serum total protein (TP) ratio and fluid ascites/serum LDH ratio, adenosine deaminase activity (ADA), and imagistic techniques were recently considered for an efficient positive diagnosis of TB ascites, making possible the early treatment with appropriate tuberculostatic drugs.
The isolation of good-quality DNA is the prerequisite for molecular research. Maintaining yield and quality of DNA during plant DNA extraction is one of the difficult tasks compared to that of animals, because of its rigid cell wall, which is made up of cellulose along with other variable levels of chemical components such as polysaccharides, polyphenols, proteins, and lipids that act as a contaminant during DNA extraction. The amount of these components varies according to plant species, plant part used, environmental condition, and growth stage and it is very problematic when isolating DNA. For example, cereals are rich in carbohydrates whereas medicinal plants are rich in the polyphenols wherein stressed plants have higher polyphenols. These contaminants can be removed during extraction by standardizing basic DNA extraction protocol [1, 2, 3].
Generally fresh leaves aged 15–20 days are preferred for plant tissues (fresh, freeze-dried, or frozen in liquid nitrogen) and usually ruptured by mechanical force in pestle and motor or TissueLyser. If liquid nitrogen is unavailable, CTAB buffer can be used directly or prewarmed for grinding. The main objective of various DNA isolation methods is development of relatively quick, inexpensive, and consistent protocol to extract high-quality DNA with better yield. Generally, leaf samples contain large quantities of polyphenols, tannins, and polysaccharides. The basic principle of DNA isolation is disruption of the cell wall, cell membrane, and nuclear membrane to release the highly intact DNA into solution followed by precipitation of DNA and removal of the contaminating biomolecules such as the proteins, polysaccharides, lipids, phenols, and other secondary metabolites by enzymatic or chemical methods [4].
The plant DNA is extracted by either CTAB-based [5, 6] or sodium dodecyl sulfate (SDS)-based methods [7]. The majority of the protocols developed for DNA extraction are modified versions of hexadecyltrimethylammonium bromide (CTAB) extraction [8]. The role of various chemicals involved in CTAB extraction method has been described in the present communication.
The CTAB buffer mainly includes CTAB, sodium chloride (NaCl), and ethylenediaminetetraacetic acid (EDTA) Tris2-amino-2-hydroxymethyl-1,3-propanediol (TRIS), polyvinylpyrrolidone (PVP), and β mercaptoethanol.
The plant cells enclose themselves in complex polysaccharide cell wall, of which cellulose is a major constituent [9], which is crystalline in nature, due to chain-like structure and intermolecular hydrogen bonding. This can be weakened to open the cell wall, by applying mechanical force exerted during grinding along with CTAB buffer or liquid nitrogen.
Cell membrane lies next to the cell wall and cellulose and is composed of a diverse set of phospholipid molecules and proteins. It dissolves in surfactant, detergents, which are amphipathic (hydrophobic tail and hydrophilic head) in nature, very much similar to phospholipid membranes. Surfactants are characterized based on their hydrophilic group, that is, ionic, nonionic, and zwitterionic. Ionic surfactant has been always better in denaturing protein molecules, and thus in dissolving the membranes [10].
CTAB, a cationic detergent, constitutes a long hydrophobic hydrocarbon chain and a hydrophilic head. It forms micelle in water because of the amphipathic nature. During DNA extraction, under aqueous condition, CTAB comes in contact with the biological membrane, captures the lipids (Figure 1), and results in the release of nucleus, which is devoid of membrane [11]. Plant tissue, which is rich in complex polysaccharides and secondary metabolites, interfere and co-precipitate with DNA; CTAB along with some other chemicals like PVP is used to minimize the effect of these metabolites.
CTAB’s role in removing membrane [
CTAB works differently based on the ionic strength of the solution. At a low ionic strength, it precipitates nucleic acid and acidic polysaccharides (pectin, xylan, and carrageenan), while protein and neutral polysaccharides (dextran, gum locust bean, starch, and inulin) remain in the solution [12]. However, at high ionic concentration, it gets bound to the polysaccharides and forms complexes that are removed during subsequent chloroform extraction. It also denatures or inhibits the activity of proteins and/or enzymes [13].
NaCl helps to remove proteins that are bound to the DNA. It also helps to keep the proteins dissolved in the aqueous layer so they do not precipitate in the alcohol along with the DNA by neutralizing the negative charges on the DNA so that the molecules can come together.
Osmosis occurs when cell is subjected to hypo or hypertonic solution. If the cells are kept in hypotonic solution, water enters inside the cell that leads to swelling, rising internal pressure and eventually bursting. On the other hand, in a hypertonic solution, water tends to ooze out from the cell and eventually plant cell shrinks and crumples, which leads to plasmolysis. Therefore, salt concentration plays a significant role in cell lysis.
The salt concentration of more than 0.5 M provides the ionic strength needed for CTAB to precipitate polysaccharides [8, 14]. In several protocols, 1.4 M concentration of NaCl has been suggested; however, in the protocols developed for getting rid of polysaccharides, higher concentration of the NaCl and/or CTAB has been recommended.
Tris is a (hydroxymethyl) aminomethane with the molecular formula (HOCH2)3CNH2, which has three primary alcohols and an amine group with a pKa of 8.1, is an effective buffer between pH 7 and 9. When the pH is adjusted to 8, with HCl, it contains a mixture of weak base and its conjugate weak acid (Figure 2), which can act as a buffer and further increases the permeability of the cell wall. When the cell wall and membranes are broken during tissue grinding, compartmentalization ends, cytoplasmic material is released, because of which the pH gets altered, and consequently the stability of biomolecules like nucleic acid is disturbed. The buffer plays a major role under such situations, and the Tris buffer maintains the pH of the solution.
Tris buffer after titration of Tris base solution [
EDTA (C10H16N2O8) chelates divalent cations, such as Mg2+ and Ca2+ (Figure 3), which is present in the enzymes and reduces the enzyme activity of DNase and RNase. Divalent cations are the cofactors for many enzymes that increase the activity of the enzyme. For example, DNase enzyme requires Mg2+ ions as a cofactor for its activity. Chelating Mg2+ ions with EDTA makes enzyme DNase nonfunctional, and thereby protects the DNA. The Mg2+ ions are also required for aggregation of nucleic acid with protein; whereas Ca2+ ions are required for cementing of cell wall’s middle layer and membrane stability. Thus, harnessing them by EDTA results in destabilization of the enzyme’s integrity.
EDTA chelates divalent cations like magnesium and calcium [
Plants are rich in phenolics compounds and to get a quality DNA these should be removed. β-Mercaptoethanol (HOCH2CH2SH) is added most of the time in extraction buffers and is a strong reducing agent to clean tannins and other polyphenols present in the crude plant extract.
Globular proteins get dissolved in water. To make them insoluble, their denaturation is one of the alternatives that can be done at tertiary and quaternary structure level of protein by reducing intermolecular disulfide linkages. β-Mercaptoethanol reduces disulfide bonds of the protein (Figure 4) and thus the proteins are denatured.
β-Mercaptoethanol reduces disulfide linkage of protein, thus denaturing it [
PVP is added to remove phenolic compounds from plant DNA extracts. Polyphenol is a major component in medicinal plants, woody plants, and mature plant parts. It is present in the vacuole, while its oxidizing enzyme, polyphenol oxidase (PPO) is located in plastid [15]. During grinding of the tissue, compartmentalization breaks and PPO convert polyphenols into quinone, which gives brown coloration. Polyphenols bind DNA and make downstream processing difficult as they get co-precipitated with the nucleic acid. PVP removes polyphenolic contamination by binding it through hydrogen bond [16, 17]. Thus, it prevents polyphenol oxidation, and thereby browning of DNA samples [18]. When the extract is centrifuged with chloroform, PVP complexes get accumulated at the interphase.
Cell lysate mixture with CTAB buffer should be kept in the water bath at 65°C, which irreversibly inhibits enzyme DNase. After removing the sample from water bath, it should be allowed to cool at room temperature, then chloroform:isoamyl alcohol (24:1) or phenol:chloroform:isoamyl alcohol (25:24:1) shall be added. Chloroform:octanol (24:1) can also be used instead of chloroform:isoamyl alcohol (24:1).
Phenol is an organic solvent, so it is not miscible with water and is used along with chloroform and isoamyl alcohol for purification of the DNA to remove proteins and polysaccharide contaminants. When phenol is shaken with cell extract, the nonpolar components of the cell will be fractionated in phenol, leaving polar ones in water. DNA is insoluble in phenol because phenol is a nonpolar solution. On the other side, protein has both polar and nonpolar groups present in it because of the long chain of different amino acids. Different amino acids have different groups present on their side chain. Also, the folding of the protein into the secondary, tertiary, and quaternary structure depends on the polarity of the amino acids. The bonds between amino acids are broken by the addition of phenol and protein gets denatured and ultimately the protein becomes unfolded.
Centrifugation after phenol:chloroform:isoamyl alcohol in 25:24:1 ratio steps gives three layers, that is aqueous, interphase, and at bottom organic phase. At neutral to alkaline pH, the nucleic acids are negatively charged and polar. Therefore, it is hydrophilic and remains in an aqueous phase. In aqueous solution, hydrophobic amino acid forms a protective core. However, after denaturation, nonpolar cores (hydrophobic) get exposed, causing precipitation of protein as well as some polysaccharides at interphase.
The phenol-chloroform combination reduces the partitioning of poly (A) and mRNA into the organic phase and reduces the formation of insoluble RNA protein complexes at the interphase. Phenol retains about 10–15% of the aqueous phase, which results in a similar loss of RNA; chloroform prevents this retention of water and thus improves yields.
Only neutral phenol should be used, as acidic phenol dissolves DNA within, or phenol turns into quinones by oxidation and it forms free radical, degrading nucleic acid. Simple observation of phenol’s pink color will state its acidic nature. The centrifugation after chloroform:isoamyl alcohol step should be done under room temperature, because below 15°C, CTAB/nucleic acid forms irreversible aggregates and may precipitate. During this step, the DNA shall be in aqueous phase [19].
Chloroform (CHCl3) or trichloromethane is a nonpolar (hydrophobic) solvent, in which nonpolar proteins and lipids get dissolved to promote the partitioning of lipids and cellular debris into the organic phase, leaving isolated DNA protected in the aqueous phase. Chloroform ensures phase separation of the two liquids because it has a higher density (1.47 g/cm3) and forces a sharper separation of the organic and aqueous phases, thereby assisting in the removal of the aqueous phase with minimal cross contamination from the organic phase. As chloroform is volatile in nature, it does not hinder the downstream process.
Chloroform comes in contact with the air and forms gas phosgene (COCl2, carbonyl chloride), which is harmful. If we simply use chloroform only, the gas entrapment causes foaming or frothing, it foams up between interphase during extraction process and makes it difficult to properly purify the DNA, which is prevented when chloroform is used along with isoamyl alcohol or isopentanol {(CH3)2CHCH2CH2OH} or octanol {CH3(CH2)7OH} by preventing the emulsification of a solution. Isoamyl alcohol or isopentanol is not miscible in the aqueous solution because it is a long-chain aliphatic compound, containing five carbon atoms and stabilizes the interphase between organic and aqueous layer. The aqueous phase contains DNA and the organic phase contains lipid, proteins, and other impurities. Isoamyl alcohol helps to inhibit RNase activity and to help prevent the solubilization in the phenol phase of long RNA molecules with long poly (A) portions. This will increase the purity of DNA.
Genomic DNA should be treated with Ribonuclease A (RNase A) to remove the contamination of RNA for DNA purification. RNase A is an endoribonuclease that catalyzes the hydrolysis of the 3′,5′-phosphodiester linkage of RNA at the 5′-ester bond in a two-step reaction. The first step is a transphosphorylation to give an oligonucleotide terminating in a pyrimidine 2′,3′-cyclic phosphate. The second is the hydrolysis of the cyclic phosphate to give a terminal 3′-phosphate. Numerous chemical studies have suggested that histidine 12, histidine 119, and lysine 41 are involved in the active site of the enzyme and the DNA is devoid of 2′OH group (deoxy), it remains secure (Figures 5 and 6) [20].
(A) The hydrolysis reaction catalyzed by RNase A. An RNA molecule is a chain of nucleotides linked by the phosphodiester bond, which may be cleaved by RNase [
The catalytic mechanism of RNase A, which contains two critical residues: His-12 and His-119 [
Alcohol is used to precipitate the DNA out of the extraction solution, so we can wash all those salts and chemicals away and then dissolve it in our final solvent—usually water or some variant of Tris-EDTA solution. DNA remains dissolved in aqueous solution because DNA has phosphodiester backbone, which is hydrophilic in nature. Water molecule forms hydration shell around DNA by forming hydrogen bonds. Isopropanol/ethanol is used in precipitation of DNA, which breaks the hydration shell. Isopropanol is a good choice for precipitation of DNA. The amount of isopropanol requirement is less (0.6–0.7 volume of supernatant), as isopropanol has a higher capacity to reduce the dielectric constant of water than the ethanol (2–3 volume) and also requires a fair amount of salt to work. RNA which has extra 2′OH remains hydrogen bounded with water more strongly than DNA tends to stay soluble in it, thus selective precipitation of DNA can be done. Isopropanol also dissolves nonpolar solvents such as chloroform, thus the impurities form previous step can also be removed.
Using ice-cold isopropanol is generally practiced, but many researchers say that it should be used at room temperature, otherwise it will precipitate polysaccharides also [21]. Though the yield of DNA will be increased at low temperature, it may increase impurities [22].
The role of the salt in the extraction protocol is to neutralize the charges on the sugar phosphate backbone of the DNA. Sodium acetate with pH 5.2 is commonly used for precipitation of nucleic acid along with ethanol [23]. In solution, sodium acetate dissociates into Na+ and [CH3COO]−. The positively charged sodium ions neutralize the negative charge on the PO3− groups on the sugar phosphate backbone of nucleic acids reducing repulsion between DNA molecules, making the DNA molecule far less hydrophilic, and therefore much less soluble in water. The electrostatic attraction between the Na+ ions in solution and the PO3− ions on the nucleic acid are dictated by Coulomb’s Law, which is affected by the dielectric constant of the solution. Water has a high dielectric constant, which makes it fairly difficult for the Na+ and PO3− to come together. This is useful in aggregation and formation of tangled mass. It is also called as salting out. Nevertheless, it is not seen when salt alone is used. It requires the solution with low dielectric constant, which allows this interaction. This is affected by either ethanol or isopropanol, which has a much lower dielectric constant, making it much easier for Na+ to interact with the PO3−, shield its charge, and make the nucleic acid less hydrophilic, causing the DNA to drop out of solution (Figure 7).
Role of salt in DNA precipitation [
DNA precipitate is washed again with 70% ethanol to rinse excess salt that might come along with the extraction buffers from the pellet [24], centrifuged, and ethanol is discarded, leaving DNA in the precipitate. Precipitate is air-dried or vacuum-dried. Over drying should be avoided as DNA converts B form to D form, which is difficult to dissolve later [25].
In older times in DNA isolation methods, DNA used to be stored dry and diluted when required. Nowadays, for long-term storage, it is prudent to store DNA in a buffer that maintains its pH and keeps it from getting degraded. TE buffer contains Tris (10 mM) and EDTA (1 mM), where Tris is the buffering component and EDTA the chelating component. For DNA isolation, the pH is usually set to 7.5–8.5, the slight alkalinity of TE buffer also prevents chances of acid hydrolysis that may further disrupt the stability of DNA stored in water. Tris amino constituent of TE buffer has the ability to protect DNA strands from radiation damage, in both solid state and fluid solution. As radiation produces free radicals, it may break DNA strands. Thus, in the fluid solution at ambient temperature Tris acts by scavenging hydroxyl radicals [26]. The purpose of EDTA is to chelate Mg2+ ions in solution necessary for DNase or RNase action, thus protecting the DNA from DNases or RNase.
Sterile water can be utilized for short-duration storage of DNA. If TE buffer is used for storage of DNA, it should be diluted further with sterile water to dilute EDTA concentration for making magnesium ions available for polymerase activity during PCR because if DNA has to be sent for sequencing afterward, the buffer components in TE hinders the process. The same EDTA that chelates ions to degrade magnesium also hinders the action of DNA polymerases during PCR, which can be overcome by adding more magnesium to the master mix, or perhaps diluting the DNA sample so that the already low concentrations of EDTA do not actually disrupt PCR. In fact, in a large number of cases, they do not.
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His studies in robotics lead him not only to a PhD degree but also inspired him to co-found and build the International Journal of Advanced Robotic Systems - world's first Open Access journal in the field of robotics.",institutionString:null,institution:{name:"TU Wien",country:{name:"Austria"}}},{id:"441",title:"Ph.D.",name:"Jaekyu",middleName:null,surname:"Park",slug:"jaekyu-park",fullName:"Jaekyu Park",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/441/images/1881_n.jpg",biography:null,institutionString:null,institution:{name:"LG Corporation (South Korea)",country:{name:"Korea, South"}}},{id:"465",title:"Dr",name:"Christian",middleName:null,surname:"Martens",slug:"christian-martens",fullName:"Christian Martens",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"479",title:"Dr.",name:"Valentina",middleName:null,surname:"Colla",slug:"valentina-colla",fullName:"Valentina Colla",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/479/images/358_n.jpg",biography:null,institutionString:null,institution:{name:"Sant'Anna School of Advanced Studies",country:{name:"Italy"}}},{id:"494",title:"PhD",name:"Loris",middleName:null,surname:"Nanni",slug:"loris-nanni",fullName:"Loris Nanni",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/494/images/system/494.jpg",biography:"Loris Nanni received his Master Degree cum laude on June-2002 from the University of Bologna, and the April 26th 2006 he received his Ph.D. in Computer Engineering at DEIS, University of Bologna. On September, 29th 2006 he has won a post PhD fellowship from the university of Bologna (from October 2006 to October 2008), at the competitive examination he was ranked first in the industrial engineering area. He extensively served as referee for several international journals. He is author/coauthor of more than 100 research papers. He has been involved in some projects supported by MURST and European Community. His research interests include pattern recognition, bioinformatics, and biometric systems (fingerprint classification and recognition, signature verification, face recognition).",institutionString:null,institution:null},{id:"496",title:"Dr.",name:"Carlos",middleName:null,surname:"Leon",slug:"carlos-leon",fullName:"Carlos Leon",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Seville",country:{name:"Spain"}}},{id:"512",title:"Dr.",name:"Dayang",middleName:null,surname:"Jawawi",slug:"dayang-jawawi",fullName:"Dayang Jawawi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Technology Malaysia",country:{name:"Malaysia"}}},{id:"528",title:"Dr.",name:"Kresimir",middleName:null,surname:"Delac",slug:"kresimir-delac",fullName:"Kresimir Delac",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/528/images/system/528.jpg",biography:"K. Delac received his B.Sc.E.E. degree in 2003 and is currentlypursuing a Ph.D. degree at the University of Zagreb, Faculty of Electrical Engineering andComputing. His current research interests are digital image analysis, pattern recognition andbiometrics.",institutionString:null,institution:{name:"University of Zagreb",country:{name:"Croatia"}}},{id:"557",title:"Dr.",name:"Andon",middleName:"Venelinov",surname:"Topalov",slug:"andon-topalov",fullName:"Andon Topalov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/557/images/1927_n.jpg",biography:"Dr. Andon V. Topalov received the MSc degree in Control Engineering from the Faculty of Information Systems, Technologies, and Automation at Moscow State University of Civil Engineering (MGGU) in 1979. He then received his PhD degree in Control Engineering from the Department of Automation and Remote Control at Moscow State Mining University (MGSU), Moscow, in 1984. 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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. Papakostas has received a diploma in Electrical and Computer Engineering in 1999 and the M.Sc. and Ph.D. degrees in Electrical and Computer Engineering in 2002 and 2007, respectively, from the Democritus University of Thrace (DUTH), Greece. Dr. Papakostas serves as a Tenured Full Professor at the Department of Computer Science, International Hellenic University, Greece. Dr. Papakostas has 10 years of experience in large-scale systems design as a senior software engineer and technical manager, and 20 years of research experience in the field of Artificial Intelligence. Currently, he is the Head of the “Visual Computing” division of HUman-MAchines INteraction Laboratory (HUMAIN-Lab) and the Director of the MPhil program “Advanced Technologies in Informatics and Computers” hosted by the Department of Computer Science, International Hellenic University. 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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