Analytical platforms for the analysis of cannabinoids in
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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.
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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.
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The strong impulse to research this topic has produced in recent years a large literature that documents the high level of complexity of the issue. Due to this complexity, a reasoned multidimensional analysis able to integrate expertise of different disciplines (neurology, neuropsychology, neuroradiology, and neuroscience) is necessary. This book offers an excellent synopsis of perspectives, methods, empirical evidences, and international references. It represents an extraordinary opportunity to target challenging unmet needs and to outline new horizons in Parkinson's disease research.",isbn:"978-1-83880-869-3",printIsbn:"978-1-83962-648-7",pdfIsbn:"978-1-83880-870-9",doi:"10.5772/intechopen.79277",price:100,priceEur:109,priceUsd:129,slug:"parkinson-s-disease-and-beyond-a-neurocognitive-approach",numberOfPages:84,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"7407cfb0a38d3c1b8dd1c578c804fc8d",bookSignature:"Sara Palermo, Mario Stanziano and Rosalba Morese",publishedDate:"June 19th 2019",coverURL:"https://cdn.intechopen.com/books/images_new/8626.jpg",numberOfDownloads:5406,numberOfWosCitations:0,numberOfCrossrefCitations:6,numberOfCrossrefCitationsByBook:1,numberOfDimensionsCitations:8,numberOfDimensionsCitationsByBook:1,hasAltmetrics:1,numberOfTotalCitations:14,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"June 4th 2018",dateEndSecondStepPublish:"September 3rd 2018",dateEndThirdStepPublish:"November 2nd 2018",dateEndFourthStepPublish:"January 21st 2019",dateEndFifthStepPublish:"March 22nd 2019",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"233998",title:"Ph.D.",name:"Sara",middleName:null,surname:"Palermo",slug:"sara-palermo",fullName:"Sara Palermo",profilePictureURL:"https://mts.intechopen.com/storage/users/233998/images/system/233998.png",biography:"Sara Palermo has an MSc in clinical psychology and a PhD in experimental neuroscience. She is specialty chief editor of Frontiers in Psychology, Neuropsychology, and scientific director of the Italian National Institute of Philanthropy, Filantropolis. She is a member of the Italian Society of Neuropsychology, the Italian Association of Psychogeriatrics, the Italian Society of Neurology for Dementia, and the Society for Interdisciplinary Placebo Studies. She was a member of the European Innovation Partnership on Active and Healthy Ageing (EIP AHA), for which she was involved in Action Group A3: Action for Prevention of Functional Decline and Frailty. Dr Palermo works as a researcher at the Department of Psychology - University of Turin (Italy) and as Scientific Consultant at the Fondazione IRCCS, Istituto Neurologico Carlo Besta (FINCB), Milan, Italy.",institutionString:"University of Turin, Italy & The Foundation of the Carlo Besta Neurological Institute IRCCS",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"6",totalChapterViews:"0",totalEditedBooks:"5",institution:{name:"University of Turin",institutionURL:null,country:{name:"Italy"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:{id:"260481",title:"Dr.",name:"Mario",middleName:null,surname:"Stanziano",slug:"mario-stanziano",fullName:"Mario Stanziano",profilePictureURL:"https://mts.intechopen.com/storage/users/260481/images/system/260481.jpg",biography:"Mario Stanziano is a physician, currently resident in Diagnostic Imaging at the University of Milan, cooperating with the Brain Imaging Center and the Trauma Center of Turin. He has been working with the neural network morphology lab of the University of Naples Vanvitelli in the Human Anatomy Department. As a neuroimager he is actively engaged in the study of normal and pathological brain connectivity both under functional and structural perspectives. In particular, he is studying structuro-functional magnetic resonance imaging features of neurological disorders characterised by consciousness impairment in its broadest sense.",institutionString:"Radiology Institute, University of Turin",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"University of Turin",institutionURL:null,country:{name:"Italy"}}},coeditorTwo:{id:"214435",title:"Dr.",name:"Rosalba",middleName:null,surname:"Morese",slug:"rosalba-morese",fullName:"Rosalba Morese",profilePictureURL:"https://mts.intechopen.com/storage/users/214435/images/system/214435.jpg",biography:"Rosalba Morese, born in Italy, holds a bachelor\\'s degree in psychology at the University of Parma and a Ph.D. in neuroscience at the University of Turin. She aims to develop new techniques and approaches in cognitive science and social neuroscience. She is an expert in experimental neuroscience, neuroeconomics, psychophysiology, and cognitive and social neuroscience. She performs neuroimaging studies in social contexts in order to investigate neural correlates involved during social interactions, such as, social exclusion, social support, empathy, communicative intention, social decision-making, in-group and out-group settings, etc. She currently works at Università della Svizzera italiana, Lugano, Switzerland.\n\nFor more information: http://usi.to/xuj",institutionString:"Faculty of Biomedical Sciences",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"8",totalChapterViews:"0",totalEditedBooks:"4",institution:{name:"Universita della Svizzera Italiana",institutionURL:null,country:{name:"Switzerland"}}},coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"1053",title:"Cognitive Psychology",slug:"cognitive-psychology"}],chapters:[{id:"67127",title:"Introductory Chapter: Targeting Unmet Needs in Parkinson’s Disease",doi:"10.5772/intechopen.86396",slug:"introductory-chapter-targeting-unmet-needs-in-parkinson-s-disease",totalDownloads:880,totalCrossrefCites:2,totalDimensionsCites:2,hasAltmetrics:0,abstract:null,signatures:"Sara Palermo, Rosalba Morese and Mario Stanziano",downloadPdfUrl:"/chapter/pdf-download/67127",previewPdfUrl:"/chapter/pdf-preview/67127",authors:[{id:"233998",title:"Ph.D.",name:"Sara",surname:"Palermo",slug:"sara-palermo",fullName:"Sara Palermo"},{id:"214435",title:"Dr.",name:"Rosalba",surname:"Morese",slug:"rosalba-morese",fullName:"Rosalba Morese"},{id:"262793",title:"Dr.",name:"Mario",surname:"Stanziano",slug:"mario-stanziano",fullName:"Mario Stanziano"}],corrections:null},{id:"64411",title:"Mitochondrial KATP Channel and Dopaminergic Vulnerability Neurons in Parkinson’s Disease",doi:"10.5772/intechopen.81862",slug:"mitochondrial-k-sub-atp-sub-channel-and-dopaminergic-vulnerability-neurons-in-parkinson-s-disease",totalDownloads:1126,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The motor deficiency control commonly characterizes Parkinson’s disease (PD), resulting in impairment of neuromuscular command, because of basal ganglia nuclei degeneration and late formation of Lewy’s bodies in the remaining dopaminergic (DA) neurons. Motor signals are triggered in high cortical motor areas and go toward the midbrain regions, where the final tuning movement takes place. PD is characterized primarily by the death of dopaminergic neurons in the regions known as substantia nigra compacta (STNc). Mutations in a couple of genes, such as Parkin1 and DJ1, correspond to the usual familial form of the disease, due to its association with oxidative stress and depolarization of mitochondrial membrane. However, this form does not explain the selective pattern of apoptosis between the neuronal dopaminergic areas of midbrain regions. In this chapter, we are putting forward the hypothesis of oxidative stress and mitochondrial changes as the apparent most relevant cause in PD, as well as the neuroprotective role played by Kir6.2, a potassium-ATP channel and calcium voltage-gated v1.3.",signatures:"Gesivaldo Santos, Julita Maria Pereira Borges, Marcos Avilla-Rodriguez, Érika Pereira Rubio, Cattiúscia Batista Bromochenkel, Djalma Menezes Oliveira, Jane Lima dos Santos, Rosane Moura Aguiar, Milena Mascarenhas Ferraz, Silvana Batista Gaino, Francisco Capani and George E. Barreto",downloadPdfUrl:"/chapter/pdf-download/64411",previewPdfUrl:"/chapter/pdf-preview/64411",authors:[{id:"120703",title:"Dr.",name:"Francisco",surname:"Capani",slug:"francisco-capani",fullName:"Francisco Capani"},{id:"125352",title:"Dr.",name:"George E.",surname:"Barreto",slug:"george-e.-barreto",fullName:"George E. Barreto"},{id:"214511",title:"Ph.D.",name:"Gesivaldo",surname:"Santos",slug:"gesivaldo-santos",fullName:"Gesivaldo Santos"},{id:"215965",title:"Dr.",name:"Marcos",surname:"Avilla-Rodriguez",slug:"marcos-avilla-rodriguez",fullName:"Marcos Avilla-Rodriguez"},{id:"278028",title:"Dr.",name:"Julita Maria Pereira",surname:"Borges",slug:"julita-maria-pereira-borges",fullName:"Julita Maria Pereira Borges"},{id:"278260",title:"Prof.",name:"Djalma Menezes",surname:"Oliveira",slug:"djalma-menezes-oliveira",fullName:"Djalma Menezes Oliveira"},{id:"278263",title:"BSc.",name:"Érika Pereira",surname:"Rubio",slug:"erika-pereira-rubio",fullName:"Érika Pereira Rubio"},{id:"278320",title:"Prof.",name:"Jane Lima",surname:"dos Santos",slug:"jane-lima-dos-santos",fullName:"Jane Lima dos Santos"},{id:"278323",title:"B.Sc.",name:"Cattiúscia",surname:"Batista Bromochenkel",slug:"cattiuscia-batista-bromochenkel",fullName:"Cattiúscia Batista Bromochenkel"},{id:"298983",title:"Dr.",name:"Rosane Moura",surname:"Aguiar",slug:"rosane-moura-aguiar",fullName:"Rosane Moura Aguiar"},{id:"298984",title:"Dr.",name:"Silvana Batista",surname:"Gaino",slug:"silvana-batista-gaino",fullName:"Silvana Batista Gaino"},{id:"298986",title:"Dr.",name:"Milena Mascarenhas",surname:"Ferraz",slug:"milena-mascarenhas-ferraz",fullName:"Milena Mascarenhas Ferraz"}],corrections:null},{id:"63549",title:"The Causative and Curative Roles of Brain-Derived Neurotrophic Factor in Parkinson’s Disease",doi:"10.5772/intechopen.81215",slug:"the-causative-and-curative-roles-of-brain-derived-neurotrophic-factor-in-parkinson-s-disease",totalDownloads:1359,totalCrossrefCites:0,totalDimensionsCites:1,hasAltmetrics:1,abstract:"Parkinson’s disease (PD) is characterized by the activation of degenerative and inflammatory processes in brain circuits that control movement and, according to the degree of progression of the damage, can cause neuropsychological disorders such as cognitive dysfunction. Changes in gene expression profile or post-translational modifications in secretory proteins such as neurotrophic factors could define the disease progression. Brain-derived neurotrophic factor (BDNF) is relevant, because it not only participates in neuronal survival, neurotransmission, dendritic growth and cellular communication but also in disease progression. In this chapter, considering both experimental evidences and clinical reports, the authors will analyze the contribution of BDNF as one of the causes of neurodegeneration and neuroinflammation; discuss the participation of this neurotrophic factor in the development of cognitive dysfunction, and finally the scope of novel BDNF-based therapies for PD.",signatures:"Daniel Hernandez-Baltazar, Rasajna Nadella, Tamara Cibrian-Llanderal, Abraham Puga-Olguín, Abril Alondra Barrientos-Bonilla, Laura Mireya Zavala-Flores, Arnulfo Villanueva-Olivo, Aurora Sanchez-Garcia, Maria de Jesús Rovirosa-Hernández and Jesus Daniel Rembao-Bojorquez",downloadPdfUrl:"/chapter/pdf-download/63549",previewPdfUrl:"/chapter/pdf-preview/63549",authors:[{id:"98494",title:"BSc.",name:"Aurora",surname:"Sánchez-García",slug:"aurora-sanchez-garcia",fullName:"Aurora Sánchez-García"},{id:"174651",title:"Dr.",name:"Abraham",surname:"Puga-Olguín",slug:"abraham-puga-olguin",fullName:"Abraham Puga-Olguín"},{id:"209886",title:"Dr.",name:"Tamara",surname:"Cibrian-Llanderal",slug:"tamara-cibrian-llanderal",fullName:"Tamara Cibrian-Llanderal"},{id:"210173",title:"Dr.",name:"Daniel",surname:"Hernandez-Baltazar",slug:"daniel-hernandez-baltazar",fullName:"Daniel Hernandez-Baltazar"},{id:"219107",title:"Dr.",name:"Rasajna",surname:"Nadella",slug:"rasajna-nadella",fullName:"Rasajna Nadella"},{id:"219108",title:"Dr.",name:"Maria De Jesus",surname:"Rovirosa-Hernandez",slug:"maria-de-jesus-rovirosa-hernandez",fullName:"Maria De Jesus Rovirosa-Hernandez"},{id:"219110",title:"Dr.",name:"Laura Mireya",surname:"Zavala-Flores",slug:"laura-mireya-zavala-flores",fullName:"Laura Mireya Zavala-Flores"},{id:"263916",title:"Dr.",name:"Abril Alondra",surname:"Barrientos-Bonilla",slug:"abril-alondra-barrientos-bonilla",fullName:"Abril Alondra Barrientos-Bonilla"},{id:"264090",title:"Dr.",name:"Arnulfo",surname:"Villanueva-Olivo",slug:"arnulfo-villanueva-olivo",fullName:"Arnulfo Villanueva-Olivo"},{id:"271563",title:"Dr.",name:"Jesus Daniel",surname:"Rembao-Bojorquez",slug:"jesus-daniel-rembao-bojorquez",fullName:"Jesus Daniel Rembao-Bojorquez"}],corrections:null},{id:"64517",title:"Neuroimaging in Parkinson Disease",doi:"10.5772/intechopen.82308",slug:"neuroimaging-in-parkinson-disease",totalDownloads:1181,totalCrossrefCites:1,totalDimensionsCites:1,hasAltmetrics:0,abstract:"Over many decades, neuroimaging which included structural, functional and molecular modalities—have provided invaluable insights into the mechanisms underlying Parkinson disease (PD). These studies have shown changes in brain structure and metabolic activity. Although it is now considered to be complex, still neuroimaging modalities are recommended for routine use in clinical practice. Special sequences such as susceptibility weighted and proton density sequences are recommended for characterization. Now, the world is switching more towards the deep brain stimulation so the neuroimaging also helps in pretreatment planning and post treatment complications assessments. This chapter discusses the radiological anatomy, sequencing and imaging appearances. It will also discuss new approaches with potential applicability to clinical practice.",signatures:"Roohi Mohammad and Fatima Mubarak",downloadPdfUrl:"/chapter/pdf-download/64517",previewPdfUrl:"/chapter/pdf-preview/64517",authors:[{id:"138863",title:"Dr.",name:"Fatima",surname:"Mubarak",slug:"fatima-mubarak",fullName:"Fatima Mubarak"},{id:"279690",title:"Dr.",name:"Roohi",surname:"Mohammad",slug:"roohi-mohammad",fullName:"Roohi Mohammad"}],corrections:null},{id:"67157",title:"Levodopa-Induced Dyskinesias and Dyskinesias-Reduced-Self-Awareness in Parkinson’s Disease: A Neurocognitive Approach",doi:"10.5772/intechopen.86384",slug:"levodopa-induced-dyskinesias-and-dyskinesias-reduced-self-awareness-in-parkinson-s-disease-a-neuroco",totalDownloads:861,totalCrossrefCites:3,totalDimensionsCites:4,hasAltmetrics:0,abstract:"Levodopa-induced dyskinesias are one of the most common disabling motor complications in advanced Parkinson’s disease. The subjective perception of motor impairment is a clinical phenomenon that needs to be adequately analyzed. Indeed, the determination of patient dyskinesias-reduced-self-awareness (DRSA) and of its relationship to daily dysfunction is an important aspect of the debate on the gold standard for treatment. As the association with executive dysfunction is a matter of debate and we hypothesize it plays an important role in DRSA, we analyzed metacognitive abilities related to action monitoring and other factors, such as response-inhibition and “Theory of Mind,” which represent a novel explanation of the phenomenon. Moreover, we investigated whether and how a dysfunction in action monitoring related to the cingulo-frontal-ventral striatal circuit would be associated with DRSA using an event-related Go-NoGo fMRI experiment. Our findings suggest the presence of executive dysfunctions in DRSA pathogenesis, with a key leading role played by the cingulo-frontal network as part of a functionally impaired response-inhibition network.",signatures:"Sara Palermo, Rosalba Morese, Carlo Alberto Artusi, Mario Stanziano and Alberto Romagnolo",downloadPdfUrl:"/chapter/pdf-download/67157",previewPdfUrl:"/chapter/pdf-preview/67157",authors:[{id:"233998",title:"Ph.D.",name:"Sara",surname:"Palermo",slug:"sara-palermo",fullName:"Sara Palermo"},{id:"214435",title:"Dr.",name:"Rosalba",surname:"Morese",slug:"rosalba-morese",fullName:"Rosalba Morese"},{id:"262793",title:"Dr.",name:"Mario",surname:"Stanziano",slug:"mario-stanziano",fullName:"Mario Stanziano"},{id:"273521",title:"Dr.",name:"Carlo Alberto",surname:"Artusi",slug:"carlo-alberto-artusi",fullName:"Carlo Alberto Artusi"},{id:"273522",title:"Dr.",name:"Alberto",surname:"Romagnolo",slug:"alberto-romagnolo",fullName:"Alberto Romagnolo"}],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},subseries:null,tags:[{id:"23",label:"women in science book program"}]},relatedBooks:[{type:"book",id:"6876",title:"Behavioral Neuroscience",subtitle:null,isOpenForSubmission:!1,hash:"61887111f05b16c8fe4aaa7826fdf39f",slug:"behavioral-neuroscience",bookSignature:"Sara Palermo and Rosalba Morese",coverURL:"https://cdn.intechopen.com/books/images_new/6876.jpg",editedByType:"Edited by",editors:[{id:"233998",title:"Ph.D.",name:"Sara",surname:"Palermo",slug:"sara-palermo",fullName:"Sara Palermo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"7821",title:"Criminology and Post-Mortem Studies",subtitle:"Analyzing Criminal Behaviour and Making Medical Decisions",isOpenForSubmission:!1,hash:"5077ee1b9a7f2a3030689f307bfb84aa",slug:"criminology-and-post-mortem-studies-analyzing-criminal-behaviour-and-making-medical-decisions",bookSignature:"Sara Palermo and Raluca Dumache",coverURL:"https://cdn.intechopen.com/books/images_new/7821.jpg",editedByType:"Edited by",editors:[{id:"233998",title:"Ph.D.",name:"Sara",surname:"Palermo",slug:"sara-palermo",fullName:"Sara Palermo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"9161",title:"Frailty in the Elderly",subtitle:"Understanding and Managing Complexity",isOpenForSubmission:!1,hash:"a4f0f2fade8fb8ba35c405f5ad31a823",slug:"frailty-in-the-elderly-understanding-and-managing-complexity",bookSignature:"Sara Palermo",coverURL:"https://cdn.intechopen.com/books/images_new/9161.jpg",editedByType:"Edited by",editors:[{id:"233998",title:"Ph.D.",name:"Sara",surname:"Palermo",slug:"sara-palermo",fullName:"Sara Palermo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"8938",title:"Inhibitory Control Training",subtitle:"A Multidisciplinary Approach",isOpenForSubmission:!1,hash:"bd82354f3bba4af5421337cd42052f86",slug:"inhibitory-control-training-a-multidisciplinary-approach",bookSignature:"Sara Palermo and Massimo Bartoli",coverURL:"https://cdn.intechopen.com/books/images_new/8938.jpg",editedByType:"Edited by",editors:[{id:"233998",title:"Ph.D.",name:"Sara",surname:"Palermo",slug:"sara-palermo",fullName:"Sara Palermo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"704",title:"Current Directions in ADHD and Its Treatment",subtitle:null,isOpenForSubmission:!1,hash:"2362fa9cf0453e26f10f1172faef1631",slug:"current-directions-in-adhd-and-its-treatment",bookSignature:"Jill M. 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\r\n\r\n\tThe book will discuss integrated management of plant diseases and plant protection, with an emphasis on phytopathogenic fungi, biocontrol, plant genetics and resistance.
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From 1998 to 2000, he presided over the implementation of the Graduate Program in Agronomy at Federal University of Uberlândia (master's and doctorate). He was director of the Institute of Agricultural Sciences at Federal University of Uberlândia from 2001 to 2004. He was president of the Brazilian Society of Phytopathology in the period from 2003 till 2004. Dr Juliatti is a member of the Brazilian Societies of Phytopathology, Plant Breeding and Brazilian Horticulture in addition to the Paulista Group of Phytopathology. He is technical consultant in the areas of corn and soybeans/ phytopathological problems in the Brazilian savannah. \nHe and the Plant Improvement team at Federal University of Uberlândia developed strains of soybean, tomato and beans with multiple resistance to phytopathogens. Together with this team, he launched 05 protected soybean cultivars for the Brazilian savannah with multiple resistance to important soybean pathosystems, including partial resistance to Soybean Asian rust. \nHe chaired the organizing committee of the Brazilian Congress on Agroenergy and the First International Symposium on Biofuel. He coordinated the Chamber of Agronomy of CREA-MG (in 2008 and 2009) and the National Coordination of Agronomy of the CREA / CONFEA system (in 2009). He is a coordinator of the Latin American Committee on studies on Sclerotinia (Sclerotinia International Working Group). Dr Juliatti is a Financial Director of SMEA (Minas Gerais Society´ of the Agronomy Engineers) and current member of the Fiscal Council of the same entity. He is also President of ABEAS (Brazilian Association of Higher Agricultural Education - Triennium 2011-2013 and 2013-2016). 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Generally, an internal standard (IS) is a nonendogenous compound, which is naturally similar to the target analyte. In this sense, this methodology is employed to increase the reproducibility of the quantification when there is a source of errors during sample analysis. These inaccuracies may be related to random and systematic errors, due to sample preparation or even the complexity of the sample, among other factors. The IS must be added to each sample in a constant amount, as well as to blank and calibration standards. In this manner, any deviation occasioned during the analysis of the sample will also affect the IS, correcting the result with the relation of both signals. For this reason, the selection of the IS is a critical step to guarantee the precision in the analysis. The stable-isotope labeled IS are the most suitable when a MS detector is used [6].
Furthermore, derivatization processes are generally required, improving the chromatographic resolution as well as peak shape of analytes [7]. For this reason, many derivatization agents have been applied, being the silylation approaches preferred. These derivatization agents are suitable to volatilize and improve mass fragmentation properties of active proton-containing groups. Commonly, N, O-bis (trimethylsilyl) trifluoroacetamide (BSTFA), N-methyl-N-(trimethylsilyl) trifluoroacetamide (MSTFA), trimethylchlorosilane (TMCS) or N-tert-butyldimethylsilyl-N- methyltrifluoroacetamide (MTBSTFA) are utilized [8]. Although this procedure turns out to be time-consuming, it is worthy since it provides increased sensitivity and reproducibility [6, 9]. Throughout this chapter, the authors will discuss the multiple analytical work modalities and methods that have been used to solve all the inconveniences found in the analysis of
Cannabinoids are the major constituents of
Cannabinoids may be classified into three groups based on their source of production,
Classification of cannabinoids based on their source of production.
Endocannabinoids (ECs) are defined as endogenous lipids that are involved in many physiological and pathological conditions, regulating neurological disorders. They are characterized by their cannabimimetic features, activating the cannabinoid receptors (CB1 and CB2) as well as other receptors. For this reason, the development of reliable methodologies to determine ECs levels is mandatory in order to understand the role of these lipid metabolites. Different ECs have been widely analyzed by GC–MS and LC–MS, like arachidonoyl ethanolamide (anandamide, AEA) and 2-arachidonyl glycerol (2-AG) in human biosamples, which are the two most widely studied [13]. Additionally, other ECs like virodhamine and noladin ether has been detected. These compounds present physiological properties comparable to natural and synthetic exogenous cannabinoids. However, the main drawback found in their determination is their low concentration, acute instability of some endogenous compounds as well as the limited sample availability. For this reason, different extraction techniques like liquid–liquid extraction (LLE) or solid phase extraction (SPE) have been used for the quantification of ECs from different samples, to increase the analyte concentration. Moreover, microextraction approaches like solid phase microextraction (SPME) have been employed in the ECs analysis, reducing the required amount of sample [14].
Several strategies using GC–MS have been described in the literature for the determination of ECs from biological samples [15, 16, 17]. In this regard, different derivatization reagents like BSTFA or methyl ester of the methoxim dimethyisopropylsilyl (DMiPSi) have been employed, in order to increase the stability of these compounds during their analysis. Additionally, different ionization modes, including electron ionization (EI), positive chemical ionization (PCI) and negative chemical ionization (NCI, also denoted as NICI, ECCI, ECNI and ECNICI) have been utilized. Such is the case of the method reported by Kayacelebi et al., based on the indirect evaluation of 2-AG by the analysis of arachidonic acid (AA) and prostaglandin E2 (PGE2), which are hydrolysis products of the EC. This approach employs GC–MS and GC–MS/MS using the NCI mode. This technique provides enhanced selectivity in comparison to EI since only analytes with high electron capture capacity or high electron affinity may be ionized, thereby eliminating potential sample matrix interferences and allowing detections at very low concentration levels (ng/L). This consideration provides an advantage in contrast to LC–MS, since sample matrix is usually one of the major issues in this methodology. In NCI, low energy electrons (around 2 eV) are produced by collision of the reagent gas (generally methane, ammonia or carbon dioxide) with electrons emitted from the filament. The resulting electrons are captured by the analyte producing stable molecular anions. Although only specified analytes fulfill the characteristics to be used by this technique, derivatization processes provide wider applicability to this unusual ionization mode. For instance, the perfluoro group presents high electron affinity, being suitable for NCI. In this manner, an esterification process of AA and PGE2 was accomplished using pentafluorobenzyl bromide (PFB-Br) as derivatization agent, allowing the controlled fragmentation of both metabolites and accordingly, resulting in a highly sensitive quantitative method [18]. Additionally, PCI has been used for the determination of AEA by GC–MS using TMS as derivatization agent. As with NCI, PCI is based on the generation of protonated molecules by collision of the reagent gas with an electron emitted from the filament. The ionized reagent gas reacts with the analyte, resulting in a positive adduct ion. In both PCI and NCI, information about the molecular weight of the analyte is provide, unlike EI where higher energy electrons collide with analytes creating fragmented positive ions and other species. Even though this methodology may appear simpler, the selectivity reached by NCI outweighs the benefits of PCI, as well as the weaknesses of the technique itself,
CANNABINOIDS | Sample treatment | Instrumental technique | Analytical features | ||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Sample | Amount of sample | Metabolite | Derivatization | Extraction | Technique | Ionization source | Column | Analysis Time (min) | LOD | LOQ | Precision (RSD, %) | Recovery (%) | Ref. | ||
Intra-assay | Inter-assay | ||||||||||||||
Plasma | 1.5 mL | AEA and 2-AG | BSTFA/TMCS | LLE | GC–MS | PCI | DB-1 MS | 16.2 | 0.3–0.5 ng/mL | 0.35–1 ng/mL | <11 | 10.6–14.1 | 42.7–72.2 | [13] | |
Blood | 0.2–5 mL | AEA and 2-AG | DMiPSi | SPE | GC–MS | EI | DB-1 | <20 | — | — | — | — | — | [15] | |
Mouse brain | — | AEA and 2-AG | BSTFA | SPE | GC–MS | PCI | Rtx-5MS | 11 | 0.01 pmol | 0.2 pmol | 2.5 | 70.9 | [16] | ||
Mouse brain | 30 mg | AEA, 2-AG and PEA | BSTFA | SPE | GC–MS | CI | CP-Sil8 CB | <30 | — | — | — | — | — | [17] | |
Dog liver | — | 2-AG (AA and PGE2) | PFB-Br | — | GC–MS and GC–MS/MS | NCI | OPTIMA-17MS | 32 | — | 10 nM / ≤0.1 μM | 2.3–12.9 | — | [18] | ||
Tissue | 50–100 mg | NAE (PEA, SEA, AEA) | BSTFA | SPE | GC–MS and GC–MS/MS | EI | Rtx-5MS | — | — | — | — | — | — | [19] | |
11-OH-THC | 0.03 pg./mg | 0.1 | 88 | [27] | |||||||||||
Hair | MSTFA | LLE | GC–MS/MS | EI | DB5-MS | 9–14 | 5–9 | [27] | |||||||
THC, CBD and CBN | 0.3–1.4 | 0.9–4.7 pg./mg | 68–97 | [27] | |||||||||||
Plant (buds, leaves and stems) | 0.2 g | CBD, THC and CBN | — | HCEE | GC-,S | EI | Zebron ZB-5MS | 30–60 | 0.05–0.09 mg/L | 0.17–0.30 mg/L | 5.6–7.4 | 97–99 | [28] | ||
5 g | THC, CBD, CBC, CBG and CBN | — | PHWE | GCXGC-QTOF-MS/MS | EI | BPX5 | 30 | — | — | 2.87–4.5 | 1.32–5.60 | 89–45–92-56 | [29] | ||
Hair | 50 mg | THC, CBD, CBN and 11-OH-THC | MSTFA | LLE | GC–MS/MS | EI | DB5-MS | 14 | 0.303–1.4 pg./mg | 0.1–4.7 pg./mg | 1.84–8.8 | 2.78–14.05 | 68–97 | [31] | |
THC-COOH | PEPA:HFIP | NCI | 8 | ||||||||||||
Serum and plasma | 1 mL | THC, THC-COOH and 11-OH-THC | MSTFA | SPE (AXS) | GC–MS | EI | OPTIMA® 5 MX Accent | <30 | 0.15–2 ng/mL | 0.3–3.3 ng/mL | 0.8–4.8 | 1.9–6.1 | 76.9–96.5 | [32] | |
Human breast milk | 0.05 mL | THC, CBN and CBD | — | SPME | HS-SPME-GC–MS | EI | HP-5MS | 16 | 10 ng/mL | 20 ng/mL | 6.5–13.3 | 2.1–9.2 | 90.9–118 | [33] | |
Hair | 25 mg | THC-COOH, OH-THC, THC, CBD and CBN | MSTFA | SPE | GC–MS/MS | EI | Zebron ZB-5MSi | 68 | 0.2–2 pg./mg | 0.5–5 pg./mg | 1.6–5.5 | 2.3–6.6 | 19–79 | [34] | |
Human serum | 1 mL | THC, THC-OH, THCA, CBD, CBDA and CBG | MSTFA | LLE | GC–MS | AP | HP-5MS | 12 | 0.05–0.9 ng/mL | 0.2–3 ng/mL | 0.2–14.6 | 6.9–14.9 | >82 | [35] | |
Plant (roots, stems, buds and leaves) | 100 mg | THCV, CBD, CBC, Δ8-THC, Δ9-THC, CBG, CBN, CBDA, THCAA and CBGA | BSTFA | — | GC-FID | — | DB-1MS | 17.5 | 0.11–0.19 μg/mL | 0.34–0.56 μg/mL | 0.19–16.79 | — | [36] | ||
Buccal swabs | 5 mg | THC, Δ8- THC, CBN, CBD, CBC, CBG, and CBDV | MSTFA | SPME | HS-SPME-GC–MS | EI | Rxi-35 Sil | 12 | <0.04 μg/mg | — | — | — | — | [37] | |
Plant (flowers) | — | CBD, THC and CBN | — | SBSE | GCXGC-QTOF-MS | EI | Rxi-5MS | 93 | 0.02–0.15 μg/mL | 0.05–0.51 μg/mL | 8.8–19.3 | — | [5] | ||
Standard mixtures | — | THCV, CBD, CBC, Δ8-THC, Δ9- THCA, 11-hydroxy-Δ9- THC and 11- nor-9- carboxy-Δ9-THC | BSTFA+1% TMCS | — | GC-UV/VUV | — | Rtx-5 | 15 | 5–10 mg/L | — | — | — | — | [11] | |
Standard mixtures | — | 62 SCs | — | — | GC–MS | PI | DB-5MS | — | — | — | — | — | — | [41] | |
Seized plant | 150 mg | 15 SCs | — | — | GC–MS | cokl-EI | DB-5MS | >15 | — | 8–133 μg/L | 2.9–7.3 | — | [42] | ||
Seized plant | — | 34 SCs | — | — | GC–MS/MS | EI | DB-5MS | >30 | 19.9–68.8 ng/mL | — | — | — | — | [43] | |
CI | 91.1–162.9 ng/mL | ||||||||||||||
Standard mixtures | 6 SCs | — | — | GC–MS | EI | Rtx-200 | 21 | — | — | — | — | — | [44] | ||
GC-IR | — | BPX5 | |||||||||||||
Plant | 10 mL | 11 SCs | — | — | GC–MS | EI PCI NCI | HP-5MS | 30 | — | — | — | — | — | [45] | |
Urine | J&W scientific | 21 | 1–5 μg/L | 5 μg/L | 6.1–16.2 | 54–98.2 | [6] | ||||||||
2 mL | 29 SCs | BSTFA | UADLLME | GC–MS | EI | 5% phenyl-methylsilicone | 1–5 μg/L | 5 μg/L | 0.5–19.7 | 93.8–105.3 | [6] | ||||
Blood | |||||||||||||||
Plant | 20 mg | 44 SCs | — | SPME | HS-SPME- | EI | DB-5 ms | 23 | — | — | — | — | — | [46] | |
Blood | 500 μL | GC–MS | |||||||||||||
Saliva | 455 μL | 5 SCs | — | MEPS | GC–MS | EI | HP-5 MS | 26 | 10–20 μg/L | 30–60 μg/L | 3.6–8.9 | 62–124 | [47] |
Analytical platforms for the analysis of cannabinoids in
Despite the fact that GC–MS has been extensively utilized to determine ECs metabolites from biological samples, in the recent years, LC–MS/MS has become the generally employed instrumentation in the analysis of ECs, being considered in this sense the reference analytical technique. This preference may be attributed to increased sensitivity and selectivity as well as high-throughput capability. In addition, derivatization processes of ECs may be avoided, which are laborious and time-consuming [20].
Phytocannabinoids are naturally occurring cannabinoids found in the
Currently, almost 150 phytocannabinoids have been detected in
For many years,
From a medical point of view, the administration of these compounds has been normally accomplished by
These compounds are metabolized in the organism after consumption, being possible the detection of their metabolites. Analysis of these chemical by-products is an excellent solution to distinguish between passive drug exposure and active consumption. In this sense, Δ9-THC is rapidly adsorbed and metabolized to 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid (Δ9-THC-COOH) and 11-hydroxy-Δ9-THC (11-THC-OH), using these compounds as
Recently, Beike et al. reported an automated process of sample preparation, which includes extraction, clean up and derivatization of cannabinoids, prior to GC-EI-MS/MS. The determination of THC as well as its metabolites Δ9-THC-COOH and 11-THC-OH in hair samples was accomplished using a SPE cartridge attached to the module of the MultiPurposeSampler (MPS) autosampler from GERSTEL, allowing the complete automation of the extraction process with relative standard deviation better than 7% [34]. In the same way, distinct GC ionization sources have been applied to achieve enhanced sensitivity, like atmospheric pressure (AP) source. This recently commercialized modality provides better ionization results than electron ionization (EI) or chemical ionization (CI) sources, enhancing selectivity and, consequently, sensitivity. AP source has been used for the quantitative determination of cannabinoids in serum samples at trace levels [35]. This softer ionization technique is based on the ionization of compounds by corona discharge in the presence of nitrogen, operating at atmospheric pressure. However, EI has been employed as the common ionization source for the analysis of
Phytocannabinoids have been widely analyzed by GC techniques, using predominantly FID and MS as detectors. One of the main shortcomings of the analysis of these compounds by GC is the direct analysis of acidic phytocannabinoids, which undergoes decarboxylation in the injector port due to the high temperatures. This drawback can be avoided by derivatization processes, which increase the chemical stability of the acidic compounds [9, 36]. For this reason, generally liquid chromatography (LC) is preferred since derivatization step is avoided. Nevertheless, greater sensitivity is reached with GC methods, still being the instrumental technique of choice. Franklin et al. reported the headspace derivatization of cannabinoids during HS-SPME step. To this end, 5 μL of derivatization reagent, MSTFA, was inserted in a 20 mL sample vial with the aid of a vial insert. The presence of MSTFA effectively derivatized the neutral cannabinoids enhancing the sensitivity of the determination. However, an earlier derivatization of the acidic cannabinoids is necessary since the performance of the headspace derivatization in this case was not so effective [37].
An additional problem associated with the determination of cannabinoids by GC–MS is the difficulty to distinguish between isomers without compromising analysis time. This is the case of Δ9-THC, which presents four isomers: Δ8 -THC, CBD, CBC, and CBL. Some strategies have been developed to overcome this problem, like the inclusion of two-dimensional GC (GCxGC) technique, which allows the identification of complex samples, identifying biomarkers and cannabinoid isomers [5]. In this case, stir bar sorptive extraction (SBSE) coated with poly-(dimethylsiloxane) (PDMS) was utilized allowing the preconcentration of a wide range of compounds with different volatilities. The retained metabolites were directly desorbed in a GCxGC–MS system, resulting in a green analytical treatment procedure. Although isomers resolution is solved with this system, analysis time remains a problem since it is necessary to perform an extraction step for at least 60 min in addition to the 93 min per run. Nevertheless, the automatization of the process may reduce human involvement and therefore systematic and random errors could be limited. Conversely, the use of GC with vacuum UV spectroscopy (VUV) could be a potential solution since differentiation among isomers may be solved in shorter analysis times (<15 min). This technology analyses compounds in the UV/VUV spectral range (120–240 nm) and is based on the excitation of chemical bonds [11]. This strategy has been utilized for the determination of different cannabinoids and their metabolites. The deconvolution of co-elution peaks allows the reduction of the chromatographic time and therefore the analysis process. However, this solution constitutes an improvement at the expense of sensitivity.
Synthetic cannabinoids (SCs) are a class of designed drugs that simulates the effects of ∆9-THC towards cannabinoid receptors. Originally, the synthesis of these compounds was intended with therapeutic purposes as a treatment of pain, being later introduced in the recreational drug market [38]. Some of the best-known SCs include AB-FUBINACA (N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-(4-fluorobenzyl)-H-indazole-3-carboxamide), AB-CHMINACA (N-(1-amino-3-methyl-1-oxobutan-2-yl)-1-(4-fluoro-benzyl)-1H-indazole-3-carboxamide), XLR-11 ((1-(5-fluoropentyl)-1H-indol-3-yl)(2,2,3,3-tetramethylcyclopropyl)methanone) and HU-210 (1,1-Dimethylheptyl-11-hydroxy-tetrahydrocannabinol), commercialized in many European and US countries as ‘Spice’, among others. Some of these cannabinoids are represented in Figure 1. There is a real risk consuming this type of drugs since their chronic abuse often leads to dangerous side effects as well as toxicity and, even in some cases, resulting in death. In addition, new psychoactive substances are constantly emerging, presenting unknown and unexpected effects. Consequently, further research is needed in this field entailing an enormous challenge, since it requires a continual actualization of the analytical techniques with the lack of available chemical reference standards. Chromatographical techniques like GC has become an important tool in this field to determine the prevalence and to assess the risks of SCs [39]. An additional problem is related to the chemical structure of these compounds, which is not well defined and therefore hinder even further their detection. While some of them, denoted as classical SCs, are structurally like natural cannabinoids, others named nonclassical and hybrid SCs present different structural features. For this reason, researchers studied fragmentation pathways of these compounds to facilitate their identification by GC–MS [40]. Although this process appears simple, some compounds result in similar mass spectra, impeding the identification of the SCs. Tandem and high-resolution MS may solve this obstacle, but generally, these equipments are not easily accessible. Therefore, more economically feasible solutions are needed. One cost-effective solution to this issue could be the utilization of different ionization modes, such as photoionization (PI). Akatsu et al. reported the utilization of GC-PI-MS for the identification of 62 SCs. This technique allows the detection of stable neutral compounds with low ionization threshold using a radical cation produced by ultraviolet light radiation which traps an electron from the target molecule. This effect is not possible with EI, which is an adequate technique for relatively high ionization threshold. In this way, with the combination of both ionization modes, it would be possible the identification of SCs [41]. Additionally, alternative ionization sources as cold-EI or CI have been described to overcome the SCs lability towards EI. These techniques increase the abundance of molecular ions allowing the discrimination between structurally-related compounds enhancing at the same time the limit of detection (LOD) and quantification (LOQ) of the procedure (see Table 1) [42, 43]. Alternatively, the combination of the GC-EI-MS and GC-IR techniques allowed the separation of six SCs regioisomers with excellent resolution results [44]. Additionally, Umebachi et al. reported the combination of GC–MS using EI, PCI and NCI for the structural elucidation and identification of different SCs in herbal products. The fragmentation of the indole and indazole SC structures was monitored with the different ionization modes, using methane as reagent gas. Although EI gives enough information about the fragmentation pattern of the molecules, PCI and NCI provide the [M+H]+ and [M-H]− fragment ions, which could be used for the molecular weight estimation [45].
On the other hand, complexity of samples may cause a reduction in the sensitivity of the determination as well as to affect the chromatographic system, particularly the stationary phase of the analytical column. Consequently, routine methods for identification of new psychoactive substances generally require quick, cheap and simple extraction methods. Scientists have been working for many years addressing this deficiency, providing new and upgraded extraction techniques to remove the undesired sample interferences. Mercieca et al. have developed a simple method to determine SCs and their metabolites in urine and blood samples. The procedure included ultrasound-assisted liquid–liquid microextraction (UALLME) and silylation derivatization coupled with GC–MS, being able to effectively identify 36 SCs and related metabolites with acceptable accuracy and precision results [7]. In the same way, HS-SPME-GC/MS has been utilized to determine 40 SCs in herbal mixtures as well as blood samples. The utilization of the SPME methodology allows the reduction of blood sample volume to 500 μL, of great advantage in the forensic analysis [46]. Recently, a new configuration using microextraction by packed sorbents (MEPS) to isolate five SCs from oral biofluids has been reported. This method allows the quantification of these compounds in the low μg/L level by GC-EI-MS, requiring just 500 μL of sample. MEPs configuration allows the potential automatization of the process for routine analysis, improving the method precision [47].
Apart from cannabinoids,
The most abundant terpenes in
Some strategies have been developed to elucidate the terpene profile in
EO | — | — | SPME | HS-SPME-GC–MS | EI | DB-5 | 60 | — | — | — | — | [51] | ||
Inflorescences and macerated oils | 100 mg | — | SPME | HS-SPME-GC–MS | EI | Rtx-Wax | 33 | — | — | — | — | [52] | ||
Inflorescences | 5 mg | — | — | SHS-GC–MS/MS | EI | DB-35MS UI | 74 | 0.001–0.123 μg/mL | 0.002–0.374 μg/mL | <11.4 | <21.9 | 81.2–119.6 | [53] | |
Plant (leaves, inflorescence and seeds) | 1 g | MSTFA:TMCS, 99:1, v/v | — | fast-GC-FID | EI | Restek RTX-5 | <16 | 0.03–0.27 μg/mL | 0.10–0.89 μg/Ml | 3.59 | 7.82 | 77.52–107.10 | [2] | |
400 | — | SPME | HS-SPME-GC–MS | EI | HP-5-cross-linked poly-5% diphenyl-95% dimethyl polysiloxane | 63 | — | — | — | — | [55] |
Analytical platforms for the analysis of terpenes in
Sample storage plays an important role in the terpene analysis, having monoterpenes a tendency to evaporate more easily than sesquiterpenes. For this reason, special attention is required when analyzing these compounds. Some strategies have been evaluated such as using frozen samples instead of dried plant material or dynamic maceration at room temperature of plant inflorescences, providing excellent results [9, 55]. Additionally, oxygenated terpenoids, because of the presence of light or oxygen, may reveal some problems during plant storage and processing. Thus, the terpene profile provides interesting information about the EO or the starting plant material, such as the aging of the product, bad storage conditions or quality and breeding conditions. Terpene analysis provides these kind of information to take into consideration [25].
As it was described before,
Seed oil | — | Fatty acids | — | SFE/SOX/PER/ULT/UTS/STU | GC–MS | EI | DB-5MS | 60 | [56] | |
Plant | 2 mg | Carbohydrates | TMCS:HMDS | LLE | GC–MS | EI | HP-5 ms | 28 | [57] | |
Plant (leaves and inflorescences) | 100 mg | Untargeted | BSTFA + TMCS | — | GC-TOF | EI | DB-5MS-UI | 53 | [50] | |
— | — | Pesticides | — | — | GC–MS/GC–MS/MS | EI | ZB- multiple residue-1 | — | [58] | |
Concentrated samples | 100 mg | Residual solvents | — | — | HS-GC–MS | EI | SHRXI- 5MS | 7 | [60] |
Analytical platforms for the analysis of other minor compounds in
In the countries where
In the cultivation of
In the analysis of products derived from
Gas chromatography is a well-established tool in the
The authors declare no conflict of interest.
A number of systemic diseases such as cardiopulmonary, infectious, inflammatory and autoimmune diseases, hematological disorders, drug toxicity and several genetic mutations lead to pulmonary hypertension (PH), a devastating disease with a high morbidity and mortality rate. Based on clinical diagnosis, PH has been classified into five major groups that were updated in 2013 [1]. Group 1, labeled as pulmonary arterial hypertension (PAH), includes idiopathic and heritable PAH, PAH associated with congenital heart defect (CHD), connective tissue diseases, portal hypertension, HIV, schistosomiasis and drug-/toxin-induced PAH. In addition, mutation of several genes such as
The major causes of PH in children are CHD, PPHN, PH associated with disruption of normal pulmonary vascular and alveolar development in preterm infants, and congenital defects associated with hypoplasia of the lungs [10, 11]. A significant number of pediatric patients (>80%) have transient PH. These include resolution of PPHN and the majority of CHD cases that become free of PAH after the surgical correction of the defect [12]. However, preterm birth itself has an increased risk of developing PH even after adjusting for known factors such as heart and lung diseases, congenital diaphragmatic hernia and chromosomal abnormalities [13]. Furthermore, poor outcome has been reported in children with
Endothelial cells (EC) play a key role in maintaining vascular homeostasis in response to various stimuli and regulate vascular tone, permeability, coagulation, inflammation through mediators such as nitric oxide (NO), endothelium-derived hyperpolarization factor (EDHF), endothelin-1 (ET1), cell adhesion molecules, cytokines and chemokines. Regardless of the underlying disease, endothelial dysfunction/disruption plays a key role in the pathogenesis of PH. The genetic and environmental factors act as an initial trigger leading to endothelial cell injury and impaired regeneration resulting in vascular remodeling and loss of small pulmonary arteries [19]. Endothelial dysfunction, impaired vascular dilatation, alterations in the expression of NO, ET1 and serotonin, increased expression of inflammatory cytokines and chemokines, loss of endothelial caveolin-1 and disordered proteolysis of extracellular matrix contribute to the pathogenesis of PAH [20, 21]. Increased expression of chemokines such as CX(3)C (fractalkine) and RANTES (CCL5) has been reported in PAH; importantly, both these chemokines are produced in EC [22, 23]. In sugen + hypoxia model (mice), the deletion of CCL5 resulted in reduction in PH via caveolin-1–dependent amplification of BMPR2 signaling. It stabilized surface caveolin-1, restored BMPR2 signaling and enhanced BMPR2 and caveolin-1 interaction [24]. This observation further supports the role for inflammation in PH. In addition, perivascular infiltration with inflammatory cells (T and B cells) is present in plexiform lesions [25, 26]. Increased expression of interleukin-1 (IL-1) and IL-6 occurs in human PAH and monocrotaline (MCT)-induced PH, and inhibition of IL-6 expression and bioactivity as a preventive measure results in the abrogation of MCT-induced PH [27, 28].
\nIn addition to the imbalance of vasoactive mediators and vascular remodeling, abnormality in ion channels (Ca2+, K+) and growth factors such as VEGF, EGF, TGF beta, MMPs, BMPR2 and Notch1 has been implicated in pathophysiology of PAH, leading to vasoconstriction, abnormal remodeling and plexiform lesions [29]. Proliferative EC reveals increased expression of angiogenesis and survival-related molecules such as VEGF, VEGFR2, Hif-1 α, and 1β and reduced expression of p27/kip1. Signal transducer and activator of transcription (STAT3) is essential for cell proliferation and survival, and antiapoptotic function [30]. In the MCT model of PH, the loss of endothelial caveolin-1 was shown to be associated with reciprocal activation of STAT3 (PY-STAT3) and increased proliferating cell nuclear antigen (PCNA) [31]. Furthermore, EC in plexiform and concentric lesions exhibits increased expression of PY-STAT3 [32]. Importantly, the inhibition of STAT3 prevents neointima formation by inhibiting cell proliferation and promoting the apoptosis of neointimal SMC [33].
Recent studies have shown the involvement of Notch1 signaling in PAH. Increased expression of Notch1 has been reported in the lungs of patients with IPAH and in rats with sugen + hypoxia-induced PH. Notch1 positively regulates EC proliferation by downregulating p21 and negatively regulating apoptosis via Bcl2 and survivin.
EC forms a monolayer in contact with blood flow and mechanical forces and underlying SMC. It is a non-thrombogenic and a selective barrier to circulating macromolecules. Juxtaposition of EC and SMC facilitates cross talk, and EC maintains SMC in quiescent state. Myoendothelial gap junction plays an important role in Ca2+ exchange between EC and SMC. EC is crucial for delivery of O2 and nutrients to underlying organs. EC maintains a balance between vasodilatation and vasoconstriction, apoptosis and cell proliferation, participate in immune and metabolic function, and maintain anticoagulant state [21, 49]. In addition, EC converts mechanical information into biological responses through mechanotransduction processes. EC adapts to mechanical inputs while maintaining crucial vascular barrier function. Failure of EC to adapt to changes has effects on vascular permeability, an important cause of vascular diseases [50].
\nCaveolae, a subset of specialized lipid rafts (50–100 nm), first described in 1950s by Palade [51] and Yamada [52], is found on plasma membranes of a variety of cell types including EC, SMC, fibroblasts and adipocytes. Caveolae are non-clathrin–coated plasma membrane vesicles (50–100 nm) enriched in glycosphingolipids, cholesterol, sphingomyelin and lipid-anchored membrane proteins. They form an important signaling platform that compartmentalizes and integrates a number of signaling molecules and allows cross talk between different signaling pathways, and mediates and integrates signaling events at the cell surface. EC contains 5000–10,000 caveolae per cell [53]. In addition, caveolae act as plasma membrane sensors and respond to stress. Caveolae flatten in response to membrane stretch. The flattening is a protective mechanism; it buffers the membrane and prevents its rupture [54, 55]. Caveolin-1 is a major protein (21–22 kDa) constituent of caveolae that maintains the shape of caveolae; EC has the highest levels of caveolin-1 [56]. Caveolin-1 is involved in multiple cellular processes such as molecular transport, cell proliferation, adhesion, migration and signal transduction. Caveolin-1 has an integral role in endocytosis. However, overexpression of caveolin-1 inhibits endocytosis [57, 58]. Caveolin-1 is synthesized in endoplasmic reticulum and then transported to Golgi complex. During its biosynthesis, it is associated with lipid rafts and become detergent resistant. From a structural standpoint, caveolin-1 contains a hairpin loop structure and three palmitoylation sites and a scaffolding domain that facilitates interaction with the plasma membrane [59, 60]. Caveolin-1 functions through protein-protein interaction and regulates and stabilizes several proteins including Src family of kinases, G proteins (α-subunits), G protein-coupled receptors, H-Ras, PKC, endothelial NO synthase (eNOS), integrins, and growth factor receptors such as VEGFR2, EGFR and PDGFR in an inhibitory conformation. Importantly, a 20-amino acid membrane proximal region of the cytosolic amino-terminal domain, termed caveolin-scaffolding domain (residue 82–101), is sufficient to mediate these interactions [61, 62]. Caveolin-1 also functions as a suppressor of cytokine signaling (SOCS), the family of proteins that are upregulated by cytokines and that in turn inhibit cytokine signaling via modulating JAK-STAT pathway [63]. Caveolin-2 is present associated with caveolin-1 in all cell types. It requires caveolin-1 for its transport from Golgi body to the plasma membrane. Caveolin-2 is not necessary for caveolae formation or caveolar localization of caveolin-1, but the coexpression results in a more efficient formation of caveolae [64]. In the absence of caveolin-1, caveolin-2 is degraded, and the decreased expression of caveolin-2 promotes increased cell proliferation [65, 66]. Furthermore, caveolin-2 knockout mice display increased proliferation of endothelial cells, hyper-cellular lung parenchyma and cell cycle progression [67].
\nIn addition to caveolin-1, caveolae require polymerase 1 and transcript release factor (PTRF) also known as cavin-1. It is an essential component of caveolae; it regulates membrane curvature by stabilizing caveolin-1 in caveolae. The loss of cavin-1 results in the loss of caveolae and the release of caveolin-1 into the plasma membrane. Importantly, caveolin-1 is required for cavin-1 recruitment to plasma membrane [68, 69]. Loss of caveolin-1 is accompanied by a marked loss of caveolin-2 and partial reduction in cavin-1 expression in the lungs. The re-expression of caveolin-1 rescues both caveolin-2 and cavin-1 [70]. In a carotid artery-injury model, the local loss of cavin-1 is reported to promote neointima formation. Furthermore, in cultured vascular SMC, the overexpression of cavin-1 suppresses SMC proliferation and migration, whereas its inhibition promotes cell proliferation and migration [71]. Cavin-1 knockout mice display lung pathological changes such as remodeled pulmonary vessels, PH and right ventricular hypertrophy. In addition, these mice have altered metabolic phenotype with insulin resistance [72, 73].
\nRecent studies have shown other accessory proteins required in caveolae biogenesis. The accessory protein pacsin2 also known as syndapin2 contains F-BAR domain associated with generation and maintenance of caveolae. It partially colocalizes with caveolin-1 at plasma membrane level. Loss of pacsin2 function results in the loss of caveolae and accumulation of caveolin-1 within the plasma membrane. Interestingly, overexpression of F-BAR domain can cause loss of caveolae. Another protein EH 15 homology domain 2 (EHD2) is present in caveolae, and it binds to pacsin2 that partially colocalizes with caveolin-1. It is a dynamin-related ATPase that confines caveolae to cell surface. Furthermore, regulation of EHD2 oligomerization in a membrane-bound state is crucial in order to restrict caveolar dynamics in cells [74, 75]. Importantly, caveolar coat controls a large number of signaling circuits; a defect in any of these pathways can lead to several systemic diseases such as vascular dysfunction, cardiomyopathy, cancer, muscular dystrophy and lipodystrophy [76].
\nThe role of caveolin-1 is well established in the pathogenesis of PH. Caveolin-1 knockout mice are viable but have dysregulated NO synthesis, impaired NO and Ca2+ signaling, cell proliferation, increased vascular permeability accompanied by cardiomyopathy and PH. Reconstituting endothelial caveolin-1 has been shown to recover dysregulated NO synthesis, cardiomyopathy and PH [77, 78]. In addition, caveolin-1 knockout mice exhibit low-grade systemic pro-inflammatory status and moderately increased IL-6 and TNFα levels [79]. EC-specific
NO, EDHF and prostacyclin (PGI2) induce endothelium-dependent vascular relaxation. NO is produced by eNOS via its action on L-arginine and oxygen. NO activates guanylate cyclase, which catalyzes the conversion of guanosine triphosphate to cyclic guanylate monophosphate. eNOS expressed in endothelial cells and cardiac myocytes is targeted to caveolae. It directly binds to caveolin-1 scaffolding domain and is held in an inhibitory state. This interaction prevents eNOS activation leading to inappropriate NO production under basal conditions. The eNOS/caveolin-1 regulatory cycle is a reversible protein-protein interaction controlled by Ca2+/calmodulin and by enzyme palmitoylation. Increase in intracellular Ca2+ with calmodulin disrupts the caveolin-1/eNOS complex resulting in eNOS activation and NO production leading to vascular relaxation. Calmodulin is a direct allosteric competitor promoting the caveolin-1 and eNOS dissociation. Heat shock protein (HSP) 90 binds to eNOS in Ca2+/calmodulin-dependent manner and it reduces the inhibitory effects of caveolin scaffolding domain on eNOS, thus promoting eNOS activation [83, 84, 85]. Furthermore, increase in vascular flow and pressure rapidly activates caveolar eNOS with its dissociation from caveolin-1 and association with calmodulin [86]. Thus, caveolin-1 and eNOS have a dynamic relationship. Importantly, caveolin-1 contained within non-caveolar lipid rafts fails to exert its inhibitory effect on eNOS [87]. The loss of endothelial caveolin-1 leads to eNOS uncoupling, oxidative stress and endothelial injury [88]. Interestingly, under conditions of stress, caveolin-1 increases eNOS trafficking in plasma membrane and primes eNOS for flow-mediated activation. Caveolin-1 plays a positive role in shear-induced eNOS activation by targeting eNOS to plasma membrane. Importantly, the coupling of flow stimulus to activate eNOS is lost in the absence of caveolin-1 and caveolae. Thus, caveolin-1 exerts dual role of post-translational regulation of eNOS activity [89]. In addition, caveolin-1 plays a critical role in VEGFR2 stimulation and downstream mediators of angiogenesis, but higher levels of caveolin-1 repress this function [90]. Interestingly, EC migration, tube formation and angiogenesis are impaired both in caveolin-1 and eNOS knockout mice but are fully restored by double knockout [91].
\nThe transient receptor potential (TRP) channels are the link between caveolae and EDHF. TRP channels facilitating the capacitive Ca2+ entry directly interact with caveolin-1 in EC. Ca2+-activated K+ channels play a key role in endothelium-dependent hyperpolarization and vascular tone regulation. Absence of caveolin-1 impairs Ca2+ homeostasis in EC and decreases the activity of TPRV4 cation channels that participate in NO and EDHF activation. Caveolin-1 is required for EDHF-related relaxation, modulating TRPV4 and connexins. Caveolin-1 knockout arteries exhibit fewer gap junctions and altered myoendothelial communication. Furthermore, caveolin-1 deficiency is associated with impaired EDHF-mediated vascular relaxation in response to shear stress and acetylcholine [92, 93, 94]. Colocalization of PGI2 synthase and caveolin-1 regulates angiogenesis [95]. Thus, caveolin-1 interacts with relaxing factors to maintain homeostasis.
\nEndothelial barrier controls the passage of fluids, nutrients and cells through vascular wall. Glycocalyx coats the luminal surface of EC and forms an important barrier. It modulates permeability, prevents leukocyte and platelet adhesion to EC, serves as an anti-inflammatory, anti-adhesive and anti-coagulant barrier, and allows selective permeability. In addition, it mediates mechanotransduction of shear stress. Under normal conditions, the apoptosis rate in EC is very low, but the activated EC exhibits a reduction in the EC surface layer, the glycocalyx, and an increased rate of apoptosis [96, 97]. Disturbed flow has been shown to inhibit glycocalyx expression as well as to reduce caveolin-1 expression in systemic arterial EC [98]. Inflammatory mediators lead to the disruption of glycocalyx resulting in the weakening of vascular protection. Integrity of vascular glycocalyx is inversely related to the degree of inflammation. Inflammatory mediators lead to the loss of glycocalyx resulting in the weakening of vascular protection. Furthermore, destruction of glycocalyx has been reported in the MCT model of PH [99].
\nCa2+-dependent vascular endothelial cadherin (VE-Cad) and its associated catenins control cell-cell adhesion and paracellular barrier function and are important for the tight junction complexes. VE-Cad is tissue specific for EC and is expressed at the intercellular clefts and mediates cell-cell adhesion, maintains barrier function, and contributes to the inhibition of cell growth. Association of caveolin-1 and VE-Cad catenin complex is essential for barrier function [100, 101, 102]. Depletion of caveolin-1 reduces VE-Cad levels and facilitates endothelial cell permeability [103]. Furthermore, VE-Cad interacts with various growth receptors, regulates endothelial proliferative signaling and mediates contact inhibition of cell growth [104, 105]. In adult EC, VE-Cad and VEGFR2 are physically linked. This maintains VEGFR2 stable and prevents its endocytosis. VEGF-induced permeability is facilitated by decoupling of VE-Cad and VEGFR2 [106]. Loss of VE-Cad and PECAM1 has been shown to occur in the MCT-induced PH [31, 107]. PECAM-1 supports EC integrity and maintains barrier function [108]. Importantly, BMPR2 also plays a role in maintaining vascular integrity by dampening inflammatory signals in pulmonary vasculature [109].
\nVascular injury from different conditions such as inflammation, hypoxia, increased flow and pressure, and shear stress leads to endothelial dysfunction. Injury can lead to disruption of EC and endothelial caveolin-1 loss or endothelial dysfunction without EC disruption. Both lead to the activation of proliferative pathways, vascular remodeling and PH [96, 110]. Recent studies have shown the loss of myocyte enhancer factor 2 (MEF2) in dysfunctional EC from PAH patients. MEF2 regulates a number of transcription factors involved in pulmonary vascular homeostasis [111]. Furthermore, these dysfunctional ECs exhibit increased production of leptin, and SMCs overexpress leptin receptor contributing to SMC proliferation [112]. In addition, pulmonary arterial ECs from PAH patients have been shown to produce increased FGF2 leading to increased proliferation and survival response by constitutive activation of ERK1/2 and decreased apoptosis associated with the activation of Bcl2 and Bcl-xL. It is thought that FGF2 in PAH may contribute to abnormal EC phenotype [113]. Furthermore, there is evidence that pro-inflammatory cytokine macrophage migration inhibitory factor and its receptor CD74 are markedly increased in idiopathic PAH, which may contribute to pro-inflammatory phenotype of EC [114].
\nEndothelial disruption accompanied by the loss of endothelial caveolin-1 has been reported in several forms of experimental models of PH such as MCT, myocardial infarction and sugen + hypoxia [31, 115, 116]. In the MCT model, progressive loss of endothelial caveolin-1 and reciprocal activation of proliferative and anti-apoptotic pathways such as PY-STAT3 and Bcl-xL occur before the onset of PH. Loss of other membrane proteins such as PECAM-1, Tie2 and soluble guanylate cyclase (α and β) occurs in tandem with caveolin-1 loss indicating extensive disruption of endothelial cell membrane. At 2 weeks, a further loss of endothelial caveolin-1 is accompanied by the loss of cytosolic proteins such as HSP90 and IκB-α and increased pulmonary artery pressure. However, at this stage, eNOS expression is relatively well preserved. In the presence of significant loss of endothelial caveolin-1 and HSP90, eNOS gets uncoupled resulting in an increased production of reactive oxygen species (ROS). By 3 and 4 weeks, there is a significant reduction in eNOS levels, leading to normalization of ROS levels [9, 31, 117]. At 4 weeks post-MCT, extensive endothelial caveolin-1 loss is accompanied by the loss of von Willebrand factor (vWF) in 29% of the arteries; and 23% of arteries exhibit enhanced expression of caveolin-1 in SMC. Enhanced expression of caveolin-1 in SMC occurs only in the arteries with extensive endothelial caveolin-1 and vWF loss. At this stage, the expression of total caveolin-1 in the lungs remains low. In addition, there is a progressive increase in MMP2 expression and activation [117]. The rescue of endothelial caveolin-1 as a preventive measure abrogates MCT-induced PH, but once the PH is established, the treatment does not alter the progression of the disease [118, 119, 120]. Exposing MCT-treated rats to hypoxia accelerates the disease process, and by 4 weeks, extensive endothelial disruption and endothelial caveolin-1 loss are accompanied by enhanced expression of caveolin-1 in SMC in 61% of the arteries, near normalization of lung caveolin-1 expression, and neointima formation. Importantly, neointimal cells exhibit low to no caveolin-1 expression [121, 122]. Extensive loss of endothelial caveolin-1, enhanced expression of caveolin-1 in SMC and neointima formation are also observed in idiopathic and hereditary PAH, PAH associated with CHD and drug toxicity [122, 123, 124, 125]. In
Apoptosis of EC in PAH is followed by proliferation of antiapoptotic EC. This concept has been confirmed in
Endothelial mesenchymal transition (EndMT) is a process by which ECs exhibit phenotype alteration. These cells lose endothelial characteristics and acquire the properties of myofibroblasts or mesenchymal cells. They exhibit loss of PECAM-1 and VE-Cad, in addition to caveolin-1, and express smooth muscle α-actin, fibroblast-specific protein 1 and Notch1. PECAM-1 and VE-Cad support EC integrity and junctional stability and maintain barrier function. Thus, their loss leads to the loss of barrier function and junction stability. These transformed ECs also acquire pro-inflammatory phenotype and are primed for proliferation, migration and tissue generation [81, 140]. Neointimal cells exhibit low levels of caveolin-1, but normal eNOS expression in the experimental model of PH and also in human PAH [96, 122], and sustained NO production has been shown to degrade caveolin-1 [141]. Importantly, caveolin-1 deficiency has been shown to induce spontaneous EndMT in pulmonary EC [142]. EndMT plays an important role in vascular remodeling, and it is also linked to the loss of BMPR2 in PH [143, 144, 145]. Furthermore, TGFβ1 plays a significant role in EndMT [146]. In addition, endothelial caveolin-1 depletion leads to eNOS uncoupling and oxidative stress that switches from BMPR2 signaling to TGFβR1 and thus may promote EndMT [80]. Plexogenic lesions contain increased VEGFA and VEGFR expression indicating misguided angiogenesis involving cells of EC origin. EC dysfunction in PAH model is shown to act through DNA methylation, histone protein modification and non-coding RNA [19]. Thus, the initial apoptosis followed by the proliferation of dysfunctional and antiapoptotic EC leads to deregulation of a number of pathways resulting in neointima and plexiform lesion formation and irreversible PAH.
\nExposure to acute hypoxia results in pulmonary arterial contraction and elevated pulmonary artery pressure, while sustained hypoxia leads to pulmonary vascular remodeling [147]. Hypoxia impairs endothelium-dependent relaxation response [148, 149]. In the MCT model of PH, the progressive loss of endothelial caveolin-1 is accompanied by a significant reduction in the expression of HSP90 (2 weeks post-MCT) and eNOS (3 weeks post-MCT) [9]. However, hypoxia does not alter the protein expression of caveolin-1, eNOS or HSP90 in the lungs. During hypoxia, caveolin-1 and eNOS have been shown to form a tight complex
People living at high altitude develop PH and right ventricular hypertrophy as an adaptive mechanism. Upon return to sea level, PH reverts to normal slowly [156]. These observations suggest that the absence of physical disruption of EC observed in the hypoxia model may be the reason why hypoxia-induced PH is reversible. Although hypoxia plays a role, inflammation and endothelial dysfunction are important factors that determine the development of PH in chronic obstructive pulmonary disease (COPD). The outflow obstruction in COPD results from inflammatory processes affecting airways, lung parenchyma and pulmonary vasculature. PH in COPD can develop independently of underlying parenchymal destruction and loss of lung vessels [157, 158]. Endothelial dysfunction has been observed in mild cases of COPD, and the loss of endothelium-dependent relaxation in the pulmonary vasculature correlates with the severity of the disease [159]. Importantly, the loss of endothelial caveolin-1 accompanied by enhanced expression of caveolin-1 in SMC is reported in COPD associated with PH. COPD without PH had preserved endothelial caveolin-1 [160]. In addition, severe pulmonary arterial lesions such as plexiform and angiomatoid lesions have been documented in explanted lungs after transplantation in COPD associated with severe PH. These lesions were similar to what are seen in IPAH [161]. In infants with respiratory distress syndrome, despite significantly elevated pulmonary artery pressure and significant medial thickening, pulmonary arteries exhibit well-preserved endothelial caveolin-1, without any evidence of EC disruption or enhanced expression of Cav-1 in SMC. In contrast, loss of endothelial Cav-1 and disruption/loss of EC coupled with enhanced expression of Cav-1 in SMC were observed in infants with bronchopulmonary dysplasia and associated inflammation [123]. These results indicate that irrespective of the underlying disease, EC disruption leads to the loss of endothelial caveolin-1 and subsequent enhanced expression of Cav-1 in SMC, followed by neointima formation and irreversible PH. Thus, the EC disruption puts the patients at a higher risk of developing irreversible PH.
\nUnder normal conditions, ECs play a key role in maintaining SMCs in quiescent state and vascular homeostasis. Caveolin-1, a major protein constituent of caveolae on the cell membrane, regulates multiple cellular processes including inflammation, molecular transport, cell proliferation, adhesion, migration and signal transduction. Caveolin-1 interacts with protein molecules that are in or are recruited to caveolae and maintains them in inhibitory confirmation. Endothelial caveolin-1 loss and caveolin-1 dysfunction lead to PH.
\nInjury such as inflammation, increased pulmonary blood flow associated with increased pressure, drugs and toxins can cause endothelial disruption, which is usually progressive. Endothelial disruption leads to the progressive loss of endothelial membrane proteins including caveolin-1, PECAM-1 and VE-Cad. These alterations lead to deregulation of multiple pathways. As depicted in Figure 1, (a) the loss of caveolin-1 is accompanied by reciprocal activation of proliferative and antiapoptotic pathways leading to SMC hypertrophy and proliferation. (b) Further loss of EC exposes SMCs to direct pressure resulting in enhanced expression of caveolin-1 in SMCs. Tyrosine phosphorylated caveolin-1 could alter the phenotype and facilitate cell migration leading to neointima formation. (c) Loss of PECAM-1 and VE-Cad results in the loss of barrier function and junction stability. These alterations lead to EndMT. These cells lose endothelial properties and acquire pro-inflammatory phenotype and are primed for proliferation, migration and tissue generation and participate in neointima formation, thus leading to irreversible PH.
\nThis figure depicts the pathway leading from endothelial cell disruption to irreversible PH. Cav-1 = caveolin-1, EC = endothelial cells, EndMT = endothelial mesenchymal transformation, PECAM-1 = platelet endothelial cell adhesion molecule 1, pCav-1 = tyrosine phosphorylated caveolin-1, PH = pulmonary hypertension, SMC = smooth muscle cells.
Hypoxia exposure to EC leads to a tight complex formation between caveolin-1 and eNOS, resulting in the dysfunction of both factors (Figure 2). Importantly, there is no EC disruption or the loss of caveolin-1 or any other membrane proteins. Since there is no loss of EC, medial layer is not exposed to shear stress and pressure. Not surprisingly, there is no enhanced expression of caveolin-1 in SMCs. However, caveolin-1 and eNOS dysfunction lead to SMC proliferation, medial hypertrophy and loss of endothelial-dependent vascular relaxation. Removal of hypoxia results in the disruption of caveolin-1/eNOS tight complex leading to reversal of PH. Slowly, the pulmonary artery pressure and medial hypertrophy return to normal as seen in experimental animals and in people returning to sea level from high altitude. Hypoxia-induced PH is reversible. However, associated inflammation/shear stress in hypoxia-induced PH, resulting in EC disruption, would lead to irreversible PH.
\nThis figure shows the effect of hypoxia on EC leading to eNOS/caveolin-1 complex formation, endothelial dysfunction and subsequent medial hypertrophy and PH. EC = endothelial cells, Cav-1 = caveolin-1, eNOS = endothelial nitric oxide synthase, SMC = smooth muscle cells, PH = pulmonary hypertension.
In conclusion, EC integrity and caveolin-1 function are important factors that determine reversible vs. irreversible PH.
\nThis work is in part supported by Cardiovascular Medical Research and Education Fund.
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Lakka and Chandrasekar Kuppan",authors:[{id:"304950",title:"Prof.",name:"Chandrasekar",middleName:null,surname:"Kuppan",slug:"chandrasekar-kuppan",fullName:"Chandrasekar Kuppan"},{id:"309984",title:"Mr.",name:"Narasimha S",middleName:null,surname:"Lakka",slug:"narasimha-s-lakka",fullName:"Narasimha S Lakka"}]},{id:"33046",title:"Affinity Chromatography: Principles and Applications",slug:"affinity-chromatography-principles-and-applications",totalDownloads:48609,totalCrossrefCites:8,totalDimensionsCites:21,abstract:null,book:{id:"1490",slug:"affinity-chromatography",title:"Affinity Chromatography",fullTitle:"Affinity Chromatography"},signatures:"Sameh Magdeldin and Annette Moser",authors:[{id:"123648",title:"Dr.",name:"Sameh",middleName:null,surname:"Magdeldin",slug:"sameh-magdeldin",fullName:"Sameh Magdeldin"},{id:"136483",title:"Dr.",name:"Annette",middleName:"C.",surname:"Moser",slug:"annette-moser",fullName:"Annette Moser"}]},{id:"50574",title:"Bioinformatics for RNA‐Seq Data Analysis",slug:"bioinformatics-for-rna-seq-data-analysis",totalDownloads:5930,totalCrossrefCites:6,totalDimensionsCites:7,abstract:"While RNA sequencing (RNA‐seq) has become increasingly popular for transcriptome profiling, the analysis of the massive amount of data generated by large‐scale RNA‐seq still remains a challenge. RNA‐seq data analyses typically consist of (1) accurate mapping of millions of short sequencing reads to a reference genome, including the identification of splicing events; (2) quantifying expression levels of genes, transcripts, and exons; (3) differential analysis of gene expression among different biological conditions; and (4) biological interpretation of differentially expressed genes. Despite the fact that multiple algorithms pertinent to basic analyses have been developed, there are still a variety of unresolved questions. In this chapter, we review the main tools and algorithms currently available for RNA‐seq data analyses, and our goal is to help RNA‐seq data analysts to make an informed choice of tools in practical RNA‐seq data analysis. In the meantime, RNA‐seq is evolving rapidly, and newer sequencing technologies are briefly introduced, including stranded RNA‐seq, targeted RNA‐seq, and single‐cell RNA‐seq.",book:{id:"5160",slug:"bioinformatics-updated-features-and-applications",title:"Bioinformatics",fullTitle:"Bioinformatics - Updated Features and Applications"},signatures:"Shanrong Zhao, Baohong Zhang, Ying Zhang, William Gordon,\nSarah Du, Theresa Paradis, Michael Vincent and David von Schack",authors:[{id:"176364",title:"Dr.",name:"Shanrong",middleName:null,surname:"Zhao",slug:"shanrong-zhao",fullName:"Shanrong Zhao"}]},{id:"49873",title:"An Introduction to Actinobacteria",slug:"an-introduction-to-actinobacteria",totalDownloads:8089,totalCrossrefCites:29,totalDimensionsCites:101,abstract:"Actinobacteria, which share the characteristics of both bacteria and fungi, are widely distributed in both terrestrial and aquatic ecosystems, mainly in soil, where they play an essential role in recycling refractory biomaterials by decomposing complex mixtures of polymers in dead plants and animals and fungal materials. They are considered as the biotechnologically valuable bacteria that are exploited for its secondary metabolite production. Approximately, 10,000 bioactive metabolites are produced by Actinobacteria, which is 45% of all bioactive microbial metabolites discovered. Especially Streptomyces species produce industrially important microorganisms as they are a rich source of several useful bioactive natural products with potential applications. Though it has various applications, some Actinobacteria have its own negative effect against plants, animals, and humans. On this context, this chapter summarizes the general characteristics of Actinobacteria, its habitat, systematic classification, various biotechnological applications, and negative impact on plants and animals.",book:{id:"5056",slug:"actinobacteria-basics-and-biotechnological-applications",title:"Actinobacteria",fullTitle:"Actinobacteria - Basics and Biotechnological Applications"},signatures:"Ranjani Anandan, Dhanasekaran Dharumadurai and Gopinath\nPonnusamy Manogaran",authors:[{id:"48914",title:"Dr.",name:"Dharumadurai",middleName:null,surname:"Dhanasekaran",slug:"dharumadurai-dhanasekaran",fullName:"Dharumadurai Dhanasekaran"}]},{id:"72074",title:"The Chemistry Behind Plant DNA Isolation Protocols",slug:"the-chemistry-behind-plant-dna-isolation-protocols",totalDownloads:3691,totalCrossrefCites:3,totalDimensionsCites:5,abstract:"Various plant species are biochemically heterogeneous in nature, a single deoxyribose nucleic acid (DNA) isolation protocol may not be suitable. There have been continuous modification and standardization in DNA isolation protocols. Most of the plant DNA isolation protocols used today are modified versions of hexadecyltrimethyl-ammonium bromide (CTAB) extraction procedure. Modification is usually performed in the concentration of chemicals used during the extraction procedure according to the plant species and plant part used. Thus, understanding the role of each chemical (viz. CTAB, NaCl, PVP, ethanol, and isopropanol) used during the DNA extraction procedure will benefit to set or modify protocols for more precisions. A review of the chemicals used in the CTAB method of DNA extraction and their probable functions on the highly evolved yet complex to students and researchers has been summarized.",book:{id:"8912",slug:"biochemical-analysis-tools-methods-for-bio-molecules-studies",title:"Biochemical Analysis Tools",fullTitle:"Biochemical Analysis Tools - Methods for Bio-Molecules Studies"},signatures:"Jina Heikrujam, Rajkumar Kishor and Pranab Behari Mazumder",authors:[{id:"74521",title:"Dr.",name:"Rajkumar",middleName:null,surname:"Kishor",slug:"rajkumar-kishor",fullName:"Rajkumar Kishor"},{id:"309357",title:"Prof.",name:"Pranab Behari",middleName:null,surname:"Mazumder",slug:"pranab-behari-mazumder",fullName:"Pranab Behari Mazumder"},{id:"318351",title:"Ph.D. Student",name:"Jina",middleName:null,surname:"Heikrujam",slug:"jina-heikrujam",fullName:"Jina Heikrujam"}]}],onlineFirstChaptersFilter:{topicId:"6",limit:6,offset:0},onlineFirstChaptersCollection:[{id:"82195",title:"Endoplasmic Reticulum: A Hub in Lipid Homeostasis",slug:"endoplasmic-reticulum-a-hub-in-lipid-homeostasis",totalDownloads:1,totalDimensionsCites:null,doi:"10.5772/intechopen.105450",abstract:"Endoplasmic Reticulum (ER) is the largest and one of the most complex cellular structures, indicating its widespread importance and variety of functions, including synthesis of membrane and secreted proteins, protein folding, calcium storage, and membrane lipid biogenesis. Moreover, the ER is implicated in cholesterol, plasmalogen, phospholipid, and sphingomyelin biosynthesis. Furthermore, the ER is in contact with most cellular organelles, such as mitochondria, peroxisomes, Golgi apparatus, lipid droplets, plasma membrane, etc. Peroxisomes are synthesized from a specific ER section, and they are related to very-long-chain fatty acid metabolism. Similarly, lipid droplets are vital structures in lipid homeostasis that are formed from the ER membrane. Additionally, there is a specific region between the ER-mitochondria interface called Mitochondria-Associated Membranes (MAMs). This small cytosolic gap plays a key role in several crucial mechanisms from autophagosome synthesis to phospholipid transfer. Due to the importance of the ER in a variety of biological processes, alterations in its functionality have relevant implications for multiple diseases. Nowadays, a plethora of pathologies like non-alcoholic steatohepatitis (NASH), cancer, and neurological alterations have been associated with ER malfunctions.",book:{id:"11674",title:"Updates on Endoplasmic Reticulum",coverURL:"https://cdn.intechopen.com/books/images_new/11674.jpg"},signatures:"Raúl Ventura and María Isabel Hernández-Alvarez"},{id:"82409",title:"Purinergic Signaling in Covid-19 Disease",slug:"purinergic-signaling-in-covid-19-disease",totalDownloads:2,totalDimensionsCites:0,doi:"10.5772/intechopen.105008",abstract:"SARS-CoV-2 virus infection causes the Covid-19 disease pandemic. Purinergic signaling is a form of extracellular signaling. Purinergic signaling plays significant role in the pathology of Covid-19. Purinergic system includes extracellular nucleotides, nucleosides, ectonucleotidases, and purinergic receptors. ATP, ADP, and adenosine are the main nucleotides, nucleosides. CD39 and CD73 are the main ectonucleotidases. There are two classes of purinergic receptors, P1 and P2. Each of them can be further divided, P1 into A1, A2A, A2B, and A3, P2 into P2X, and P2Y. In Covid-19, the purinergic system is disordered. SARS-CoV-2 viruses invading leads to extracellular ATP and ADP accumulation, purinergic receptor abnormally activation, tissue homeostasis balance is broken, which lead to inflammation even hyperinflammation with cytokine storm and thrombosis et al. symptoms. Currently, Covid-19 therapeutic medicine is still in shortage. Target purinergic system components is a promising way to treat Covid-19, which will help inhibit inflammation and prevent thrombosis. Currently, many relevant preclinical and clinical trials are ongoing. Some are very promising.",book:{id:"10801",title:"Purinergic System",coverURL:"https://cdn.intechopen.com/books/images_new/10801.jpg"},signatures:"Hailian Shen"},{id:"81708",title:"High Throughput Methods to Transfer DNA in Cells and Perspectives",slug:"high-throughput-methods-to-transfer-dna-in-cells-and-perspectives",totalDownloads:5,totalDimensionsCites:0,doi:"10.5772/intechopen.104542",abstract:"Genome sequencing led to thousands of genes to study and their molecular cloning to provide ORF collection plasmids. The main approach to study their function involves analysis of the biological consequences of their expression or knockdown, in a cellular context. Given that, the starting point of such experiments is the delivery of the exogenous material, including plasmid DNA in cells. During the last decades, efforts were made to develop efficient methods and protocols to achieve this goal. The present chapter will first give a rapid overview of the main DNA transfer methods described so far: physical, chemical, and biological. Secondly, it will focus on the different methods having reached high-throughput nowadays. Finally, it will discuss the perspectives of this field in terms of future enhancements.",book:{id:"11356",title:"Molecular Cloning",coverURL:"https://cdn.intechopen.com/books/images_new/11356.jpg"},signatures:"Colin Béatrice and Couturier Cyril"},{id:"82374",title:"The Potential of the Purinergic System as a Therapeutic Target of Natural Compounds in Cutaneous Melanoma",slug:"the-potential-of-the-purinergic-system-as-a-therapeutic-target-of-natural-compounds-in-cutaneous-mel",totalDownloads:7,totalDimensionsCites:0,doi:"10.5772/intechopen.105457",abstract:"Cutaneous melanoma is an aggressive and difficult-to-treat disease that has rapidly grown worldwide. The pharmacotherapy available in so many cases results in low response and undesirable side effects, which impair the life quality of those affected. Several studies have been shown that the purinergic system is involved in cancer context, such as in cutaneous melanoma. With technological advances, several bioactive compounds from nature are studied and presented as promising adjuvant therapies against cancer, as phenolic compounds and related action by purinergic system modulations. Thus, phenolic compounds such as rosmarinic acid, resveratrol, tannic acid, as well as vitamin D may be promising substances in a therapeutic perspective to treat cutaneous melanoma via purinergic system pathway. More research needs to be done to open up new horizons in the treatment of melanoma by the purinergic signaling.",book:{id:"10801",title:"Purinergic System",coverURL:"https://cdn.intechopen.com/books/images_new/10801.jpg"},signatures:"Gilnei Bruno da Silva, Daiane Manica, Marcelo Moreno and Margarete Dulce Bagatini"},{id:"82338",title:"Advantages of Noncoding RNAs in Molecular Diagnosis",slug:"advantages-of-noncoding-rnas-in-molecular-diagnosis",totalDownloads:7,totalDimensionsCites:0,doi:"10.5772/intechopen.105525",abstract:"Noncoding RNAs contribute to physiological processes by regulating many intracellular molecules participating in the life-supporting mechanisms of development, differentiation, and regeneration as well as by disrupting various signaling mechanisms such as disease development and progression and tumor growth. Because microRNAs (miRNAs) target and regulate the functions of key proteins, it is very useful to identify specific miRNAs that contribute to cellular functions and to clarify the roles of their target molecules as diagnostic and therapeutic strategies for cancer prognosis and treatment. In this section, the roles of miRNAs in various cancers and the processes leading to the identification of their target molecules are described, and the latest diagnostic strategies using miRNAs are discussed with specific examples.",book:{id:"11353",title:"Recent Advances in Non-Coding RNAs",coverURL:"https://cdn.intechopen.com/books/images_new/11353.jpg"},signatures:"Tomomi Fujii, Tomoko Uchiyama and Maiko Takeda"},{id:"82298",title:"Predicting SNPs in Mature MicroRNAs Dysregulated in Breast Cancer",slug:"predicting-snps-in-mature-micrornas-dysregulated-in-breast-cancer",totalDownloads:8,totalDimensionsCites:0,doi:"10.5772/intechopen.105514",abstract:"Breast cancer (BC) is the leading type of cancer among women. Findings have revolutionized current knowledge of microRNA (miRNA) in breast tumorigenesis. The seed region of miRNA regulates the process of gene expression negatively. The presence of SNPs in the seed regions of miRNA dramatically alters the mature miRNA function. Additionally, SNPs in the out-seed region of miRNAs have a significant impact on miRNA targeting. This study focuses on the in silico analysis procedure of mature miRNA SNPs and their impact on BC risk. The database annotated SNPs on mature miRNAs was used. Also, target gene alterations, miRNAs function in BC, and the interaction of miRNAs with targets were predicted. A list of 101 SNPs in 100 miRNAs with functional targets in BC was indicated. Under the SNPs allele variation, 10 miRNAs changed function, 6 miRNAs lost targets, 15 miRNAs gained targets, 48 onco-miRNAs remained unchanged, and 21 tumor suppressor miRNAs remained unchanged. At last, a list of 89 SNPs, which alter miRNA function and miRNA-mRNA interaction, were shown to be potentially associated with BC risk. This research theoretically generated a list of possible causative SNPs in the mature miRNA gene that might be used in future BC management studies.",book:{id:"11353",title:"Recent Advances in Non-Coding RNAs",coverURL:"https://cdn.intechopen.com/books/images_new/11353.jpg"},signatures:"Thanh Thi Ngoc Nguyen, Thu Huynh Ngoc Nguyen, Luan Huu Huynh, Hoang Ngo Phan and Hue Thi Nguyen"}],onlineFirstChaptersTotal:59},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:0,limit:8,total:null},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:32,numberOfPublishedChapters:318,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:12,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:19,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:15,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{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"}}}},{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"}}}}]},series:{item:{id:"3",title:"Dentistry",doi:"10.5772/intechopen.71199",issn:"2631-6218",scope:"\r\n\tThis book series will offer a comprehensive overview of recent research trends as well as clinical applications within different specialties of dentistry. Topics will include overviews of the health of the oral cavity, from prevention and care to different treatments for the rehabilitation of problems that may affect the organs and/or tissues present. The different areas of dentistry will be explored, with the aim of disseminating knowledge and providing readers with new tools for the comprehensive treatment of their patients with greater safety and with current techniques. Ongoing issues, recent advances, and future diagnostic approaches and therapeutic strategies will also be discussed. This series of books will focus on various aspects of the properties and results obtained by the various treatments available, whether preventive or curative.
",coverUrl:"https://cdn.intechopen.com/series/covers/3.jpg",latestPublicationDate:"May 13th, 2022",hasOnlineFirst:!0,numberOfPublishedBooks:8,editor:{id:"419588",title:"Ph.D.",name:"Sergio",middleName:"Alexandre",surname:"Gehrke",slug:"sergio-gehrke",fullName:"Sergio Gehrke",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038WgMKQA0/Profile_Picture_2022-06-02T11:44:20.jpg",biography:"Dr. Sergio Alexandre Gehrke is a doctorate holder in two fields. The first is a Ph.D. in Cellular and Molecular Biology from the Pontificia Catholic University, Porto Alegre, Brazil, in 2010 and the other is an International Ph.D. in Bioengineering from the Universidad Miguel Hernandez, Elche/Alicante, Spain, obtained in 2020. In 2018, he completed a postdoctoral fellowship in Materials Engineering in the NUCLEMAT of the Pontificia Catholic University, Porto Alegre, Brazil. He is currently the Director of the Postgraduate Program in Implantology of the Bioface/UCAM/PgO (Montevideo, Uruguay), Director of the Cathedra of Biotechnology of the Catholic University of Murcia (Murcia, Spain), an Extraordinary Full Professor of the Catholic University of Murcia (Murcia, Spain) as well as the Director of the private center of research Biotecnos – Technology and Science (Montevideo, Uruguay). Applied biomaterials, cellular and molecular biology, and dental implants are among his research interests. He has published several original papers in renowned journals. In addition, he is also a Collaborating Professor in several Postgraduate programs at different universities all over the world.",institutionString:null,institution:{name:"Universidad Católica San Antonio de Murcia",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:2,paginationItems:[{id:"1",title:"Oral Health",coverUrl:"https://cdn.intechopen.com/series_topics/covers/1.jpg",isOpenForSubmission:!0,annualVolume:11397,editor:{id:"173955",title:"Prof.",name:"Sandra",middleName:null,surname:"Marinho",slug:"sandra-marinho",fullName:"Sandra Marinho",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRGYMQA4/Profile_Picture_2022-06-01T13:22:41.png",biography:"Dr. Sandra A. Marinho is an Associate Professor and Brazilian researcher at the State University of Paraíba (Universidade Estadual da Paraíba- UEPB), Campus VIII, located in Araruna, state of Paraíba since 2011. She holds a degree in Dentistry from the Federal University of Alfenas (UNIFAL), while her specialization and professional improvement in Stomatology took place at Hospital Heliopolis (São Paulo, SP). Her qualifications are: a specialist in Dental Imaging and Radiology, Master in Dentistry (Periodontics) from the University of São Paulo (FORP-USP, Ribeirão Preto, SP), and Doctor (Ph.D.) in Dentistry (Stomatology Clinic) from Hospital São Lucas of the Pontifical Catholic University of Rio Grande do Sul (HSL-PUCRS, Porto Alegre, RS). She held a postdoctoral internship at the Federal University from Jequitinhonha and Mucuri Valleys (UFVJM, Diamantina, MG). She is currently a member of the Brazilian Society for Dental Research (SBPqO) and the Brazilian Society of Stomatology and Pathology (SOBEP). Dr. Marinho's experience in Dentistry mainly covers the following subjects: oral diagnosis, oral radiology; oral medicine; lesions and oral infections; oral pathology, laser therapy and epidemiological studies.",institutionString:null,institution:{name:"State University of Paraíba",institutionURL:null,country:{name:"Brazil"}}},editorTwo:null,editorThree:null},{id:"2",title:"Prosthodontics and Implant Dentistry",coverUrl:"https://cdn.intechopen.com/series_topics/covers/2.jpg",isOpenForSubmission:!0,annualVolume:11398,editor:{id:"179568",title:"Associate Prof.",name:"Wen Lin",middleName:null,surname:"Chai",slug:"wen-lin-chai",fullName:"Wen Lin Chai",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRHGAQA4/Profile_Picture_2022-05-23T14:31:12.png",biography:"Professor Dr. Chai Wen Lin is currently a lecturer at the Department of Restorative Dentistry, Faculty of Dentistry of the University of Malaya. She obtained a Master of Dental Science in 2006 and a Ph.D. in 2011. Her Ph.D. research work on the soft tissue-implant interface at the University of Sheffield has yielded several important publications in the key implant journals. She was awarded an Excellent Exchange Award by the University of Sheffield which gave her the opportunity to work at the famous Faculty of Dentistry of the University of Gothenburg, Sweden, under the tutelage of Prof. Peter Thomsen. In 2016, she was appointed as a visiting scholar at UCLA, USA, with attachment in Hospital Dentistry, and involvement in research work related to zirconia implant. In 2016, her contribution to dentistry was recognized by the Royal College of Surgeon of Edinburgh with her being awarded a Fellowship in Dental Surgery. She has authored numerous papers published both in local and international journals. She was the Editor of the Malaysian Dental Journal for several years. Her main research interests are implant-soft tissue interface, zirconia implant, photofunctionalization, 3D-oral mucosal model and pulpal regeneration.",institutionString:null,institution:{name:"University of Malaya",institutionURL:null,country:{name:"Malaysia"}}},editorTwo:{id:"479686",title:"Dr.",name:"Ghee Seong",middleName:null,surname:"Lim",slug:"ghee-seong-lim",fullName:"Ghee Seong Lim",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00003ScjLZQAZ/Profile_Picture_2022-06-08T14:17:06.png",biography:"Assoc. Prof Dr. Lim Ghee Seong graduated with a Bachelor of Dental Surgery from University of Malaya, Kuala Lumpur in 2008. He then pursued his Master in Clinical Dentistry, specializing in Restorative Dentistry at Newcastle University, Newcastle, UK, where he graduated with distinction. He has also been awarded the International Training Fellowship (Restorative Dentistry) from the Royal College of Surgeons. His passion for teaching then led him to join the faculty of dentistry at University Malaya and he has since became a valuable lecturer and clinical specialist in the Department of Restorative Dentistry. He is currently the removable prosthodontic undergraduate year 3 coordinator, head of the undergraduate module on occlusion and a member of the multidisciplinary team for the TMD clinic. He has previous membership in the British Society for Restorative Dentistry, the Malaysian Association of Aesthetic Dentistry and he is currently a lifetime member of the Malaysian Association for Prosthodontics. Currently, he is also the examiner for the Restorative Specialty Membership Examinations, Royal College of Surgeons, England. He has authored and co-authored handful of both local and international journal articles. 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Thus proteomics, an area of research that detects all protein forms expressed in an organism, including splice isoforms and post-translational modifications, is more suitable than genomics for a comprehensive understanding of the biochemical processes that govern life. The most common proteomics applications are currently in the clinical field for the identification, in a variety of biological matrices, of biomarkers for diagnosis and therapeutic intervention of disorders. From the comparison of proteomic profiles of control and disease or different physiological states, which may emerge, changes in protein expression can provide new insights into the roles played by some proteins in human pathologies. Understanding how proteins function and interact with each other is another goal of proteomics that makes this approach even more intriguing. Specialized technology and expertise are required to assess the proteome of any biological sample. Currently, proteomics relies mainly on mass spectrometry (MS) combined with electrophoretic (1 or 2-DE-MS) and/or chromatographic techniques (LC-MS/MS). MS is an excellent tool that has gained popularity in proteomics because of its ability to gather a complex body of information such as cataloging protein expression, identifying protein modification sites, and defining protein interactions. 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