\\n\\n
Dr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\\n\\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\\n\\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\\n\\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\\n\\nThank you all for being part of the journey. 5,000 times thank you!
\\n\\nNow with 5,000 titles available Open Access, which one will you read next?
\\n\\nRead, share and download for free: https://www.intechopen.com/books
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Preparation of Space Experiments edited by international leading expert Dr. Vladimir Pletser, Director of Space Training Operations at Blue Abyss is the 5,000th Open Access book published by IntechOpen and our milestone publication!
\n\n"This book presents some of the current trends in space microgravity research. The eleven chapters introduce various facets of space research in physical sciences, human physiology and technology developed using the microgravity environment not only to improve our fundamental understanding in these domains but also to adapt this new knowledge for application on earth." says the editor. Listen what else Dr. Pletser has to say...
\n\n\n\nDr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\n\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\n\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\n\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\n\nThank you all for being part of the journey. 5,000 times thank you!
\n\nNow with 5,000 titles available Open Access, which one will you read next?
\n\nRead, share and download for free: https://www.intechopen.com/books
\n\n\n\n
\n'}],latestNews:[{slug:"webinar-introduction-to-open-science-wednesday-18-may-1-pm-cest-20220518",title:"Webinar: Introduction to Open Science | Wednesday 18 May, 1 PM CEST"},{slug:"step-in-the-right-direction-intechopen-launches-a-portfolio-of-open-science-journals-20220414",title:"Step in the Right Direction: IntechOpen Launches a Portfolio of Open Science Journals"},{slug:"let-s-meet-at-london-book-fair-5-7-april-2022-olympia-london-20220321",title:"Let’s meet at London Book Fair, 5-7 April 2022, Olympia London"},{slug:"50-books-published-as-part-of-intechopen-and-knowledge-unlatched-ku-collaboration-20220316",title:"50 Books published as part of IntechOpen and Knowledge Unlatched (KU) Collaboration"},{slug:"intechopen-joins-the-united-nations-sustainable-development-goals-publishers-compact-20221702",title:"IntechOpen joins the United Nations Sustainable Development Goals Publishers Compact"},{slug:"intechopen-signs-exclusive-representation-agreement-with-lsr-libros-servicios-y-representaciones-s-a-de-c-v-20211123",title:"IntechOpen Signs Exclusive Representation Agreement with LSR Libros Servicios y Representaciones S.A. de C.V"},{slug:"intechopen-expands-partnership-with-research4life-20211110",title:"IntechOpen Expands Partnership with Research4Life"},{slug:"introducing-intechopen-book-series-a-new-publishing-format-for-oa-books-20210915",title:"Introducing IntechOpen Book Series - A New Publishing Format for OA Books"}]},book:{item:{type:"book",id:"2010",leadTitle:null,fullTitle:"Performance Evaluation of Bearings",title:"Performance Evaluation of Bearings",subtitle:null,reviewType:"peer-reviewed",abstract:"Bearings (both plain and rolling element) are used as important supporting elements for locating rotating components and confining their motion in desired direction. In order to ensure their operational reliability and desired life, these need to be properly designed/selected for an application more so because of ever increasing operational speeds. This requires the careful performance evaluation of different types of bearings considering aspects such as thermal stability, lubrication, contaminants in lubricants and controlling mechanism etc. The title of this book was specifically chosen as Performance Evaluation of Bearings. The present book is a compilation of different aspects contributing towards the performance evaluation of plain bearings (both journal and thrust), rolling element bearings and magnetic bearings.",isbn:null,printIsbn:"978-953-51-0786-6",pdfIsbn:"978-953-51-6243-8",doi:"10.5772/2421",price:119,priceEur:129,priceUsd:155,slug:"performance-evaluation-of-bearings",numberOfPages:250,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"e0f92ae90f319916c124ac47e6bbeaaf",bookSignature:"Rakesh Sehgal",publishedDate:"October 3rd 2012",coverURL:"https://cdn.intechopen.com/books/images_new/2010.jpg",numberOfDownloads:29225,numberOfWosCitations:5,numberOfCrossrefCitations:5,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:7,numberOfDimensionsCitationsByBook:0,hasAltmetrics:0,numberOfTotalCitations:17,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"December 5th 2011",dateEndSecondStepPublish:"January 9th 2012",dateEndThirdStepPublish:"April 14th 2012",dateEndFourthStepPublish:"July 13th 2012",dateEndFifthStepPublish:"August 12th 2012",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6,7",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"136404",title:"Dr.",name:"Rakesh",middleName:null,surname:"Sehgal",slug:"rakesh-sehgal",fullName:"Rakesh Sehgal",profilePictureURL:"https://mts.intechopen.com/storage/users/136404/images/3579_n.jpg",biography:"Rakesh Sehgal is currently Professor at the Department of Mechanical Engineering and Dean (Faculty Welfare) at National Institute of Technology, Hamirpur, Himachal Pradesh, India. 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Enabling Technologies and Emerging Applications",slug:"medical-internet-of-things-m-iot-enabling-technologies-and-emerging-applications",publishedDate:"February 27th 2019",bookSignature:"Hamed Farhadi",coverURL:"https://cdn.intechopen.com/books/images_new/6655.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",editors:[{id:"171143",title:"Dr.",name:"Hamed",middleName:null,surname:"Farhadi",slug:"hamed-farhadi",fullName:"Hamed Farhadi"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}}},ofsBook:{item:{type:"book",id:"11796",leadTitle:null,title:"Cytomegalovirus - Recent Advances",subtitle:null,reviewType:"peer-reviewed",abstract:"
\r\n\tCytomegalovirus (CMV) is a serious infection virus that is related to the herpes viruses and causes chickenpox and mononucleosis. CMV can develop in people who are immunocompromised and immunodeficient, called opportunistic infections. Opportunistic infections only occur if your immune system is quite weakened. Most adults carry CMV but are unaware of it because the virus cannot produce disease. In people with severely weakened immune systems, CMV can make a person feel as though they have mono. CMV can also cause serious diseases in different parts of the body. CMV spreads from person to person through body fluids, such as blood, saliva, urine, semen, and breast milk. Women who develop an active CMV infection during pregnancy can pass the virus to their neonates, who might then experience symptoms. Human cytomegalovirus (HCMV) infection induces both innate immune responses including Natural Killer cells as well as adaptive humoral and cell-mediated (CD4+ helper, CD8+ cytotoxic and γδ T cell) responses which lead to the resolution of acute primary infection. Therefore, recognition of immunopathology, pathogenesis and immune response against CMV are necessary.
",isbn:"978-1-80356-756-3",printIsbn:"978-1-80356-755-6",pdfIsbn:"978-1-80356-757-0",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,isNomenclature:!1,hash:"4e442adc2808f68ccc1aeac17e6ae746",bookSignature:"Dr. Seyyed Shamsadin Athari and Dr. Entezar Mehrabi Nasab",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11796.jpg",keywords:"Cytomegalovirus, Herpesvirales, Pathogenesis, Immunogenesis, CMV, dsDNA, HCMV, HHV-5, Immunocompromised, AIDS, Immunology, Pathogenesis",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 25th 2022",dateEndSecondStepPublish:"June 2nd 2022",dateEndThirdStepPublish:"August 1st 2022",dateEndFourthStepPublish:"October 20th 2022",dateEndFifthStepPublish:"December 19th 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"a month",secondStepPassed:!0,areRegistrationsClosed:!1,currentStepOfPublishingProcess:3,editedByType:null,kuFlag:!1,biosketch:"He has published more than 90 manuscripts in international journals on immunology, allergy, and asthma and more than 28 books. He is also on the editorial board of more than 65 international journals in medical sciences and has more than 12 inventions in medical sciences and has recorded 12 gene sequences in the gene bank. Dr. Athari has been invited as a top speaker for more than 40 international congresses and symposiums and has received several scientific awards and has 13 patents.",coeditorOneBiosketch:"Dr. Mehrebi Nasab has published more than 10 scientific articles in international journals and also she has been invited as a speaker in more than 15 international congresses. She has received some scientific awards from different scientific societies.",coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"139889",title:"Dr.",name:"Seyyed Shamsadin",middleName:null,surname:"Athari",slug:"seyyed-shamsadin-athari",fullName:"Seyyed Shamsadin Athari",profilePictureURL:"https://mts.intechopen.com/storage/users/139889/images/system/139889.jpeg",biography:"Dr. Seyyed Shamsadin Athari, MPH, Ph.D., is an Assistant Professor of Immunology, Department of Immunology, School of medicine, Zanjan University of Medical Sciences, Iran. He completed postdocs in allergy and asthma toxicology and, asthma management and controlling a network fellowship. He has published more than 30 books and 110 papers in international journals in immunology, allergy, and asthma. He is also on the editorial board of more than seventy journals. He has several scientific inventions to his credit and has recorded gene sequences in a gene bank. Dr. Athari has been invited as a top speaker at more than forty international congresses and symposiums. He is a recipient of several top researcher and young scientist awards.",institutionString:"Zanjan University of Medical Sciences",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Zanjan University of Medical Sciences",institutionURL:null,country:{name:"Iran"}}}],coeditorOne:{id:"288617",title:"Dr.",name:"Entezar Mehrabi",middleName:null,surname:"Nasab",slug:"entezar-mehrabi-nasab",fullName:"Entezar Mehrabi Nasab",profilePictureURL:"https://mts.intechopen.com/storage/users/288617/images/system/288617.jpeg",biography:"Dr. Entezar Mehrabi Nasab, MD, is a cardiologist specializing in cardiovascular diseases who completed her residency at the Tehran Heart Center, Tehran University of Medical Sciences, Iran. She has published more than twenty scientific articles in international journals. She has been an invited speaker at more than twenty international congresses. 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From chapter submission and review to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"72",title:"Ionic Liquids",subtitle:"Theory, Properties, New Approaches",isOpenForSubmission:!1,hash:"d94ffa3cfa10505e3b1d676d46fcd3f5",slug:"ionic-liquids-theory-properties-new-approaches",bookSignature:"Alexander Kokorin",coverURL:"https://cdn.intechopen.com/books/images_new/72.jpg",editedByType:"Edited by",editors:[{id:"19816",title:"Prof.",name:"Alexander",surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"2270",title:"Fourier Transform",subtitle:"Materials Analysis",isOpenForSubmission:!1,hash:"5e094b066da527193e878e160b4772af",slug:"fourier-transform-materials-analysis",bookSignature:"Salih Mohammed Salih",coverURL:"https://cdn.intechopen.com/books/images_new/2270.jpg",editedByType:"Edited by",editors:[{id:"111691",title:"Dr.Ing.",name:"Salih",surname:"Salih",slug:"salih-salih",fullName:"Salih Salih"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"117",title:"Artificial Neural Networks",subtitle:"Methodological Advances and Biomedical Applications",isOpenForSubmission:!1,hash:null,slug:"artificial-neural-networks-methodological-advances-and-biomedical-applications",bookSignature:"Kenji Suzuki",coverURL:"https://cdn.intechopen.com/books/images_new/117.jpg",editedByType:"Edited by",editors:[{id:"3095",title:"Prof.",name:"Kenji",surname:"Suzuki",slug:"kenji-suzuki",fullName:"Kenji Suzuki"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3828",title:"Application of Nanotechnology in Drug Delivery",subtitle:null,isOpenForSubmission:!1,hash:"51a27e7adbfafcfedb6e9683f209cba4",slug:"application-of-nanotechnology-in-drug-delivery",bookSignature:"Ali Demir Sezer",coverURL:"https://cdn.intechopen.com/books/images_new/3828.jpg",editedByType:"Edited by",editors:[{id:"62389",title:"PhD.",name:"Ali Demir",surname:"Sezer",slug:"ali-demir-sezer",fullName:"Ali Demir Sezer"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"40457",title:"Dispersion Relations and Modal Patterns of Wave in a Cylindrical Shell",doi:"10.5772/50477",slug:"dispersion-relations-and-modal-patterns-of-wave-in-a-cylindrical-shell",body:'The dispersion relation of a circular cylindrical shell had been a subject of great interest for several decades [1-3]. In some earlier works, numerical procedures were used exclusively in the computation of dispersion relations. Recently, Karczub [4] obtained an analytical expression for the dispersion relations from Flügge shell theory by using a symbolic algebra package, Mathematica. Karczub’s main concern was to check the agreement between his analytical results and results obtained previously from numerical methods, and limited to the harmonic orders
The solutions of the modal patterns for each of the propagating and non-propagating modes are also important and are not discussed in Karczub’s paper. These solutions are crucial to determine the vibration of a finite shell under various admissible boundary conditions and arbitrary external forces. The dispersion relations and the associated eigenvectors are also the means by which to construct transfer matrices for vibroacoustic transmission in cylindrical shell structures or pipe-hose systems [5]. The traditional and standard method of finding eigenvectors was given in detail by Leissa [6]; the eigenvector was normalized in such a way that its radial component of the displacement was unity, while the longitudinal and circumferential components were expressed as displacement ratios relative to the radial displacement. In some cases, these ratios could become exceedingly large and the eigenvector thus could deviate significantly from the eigenvector commonly used in mathematical physics.
In order to improve this problem for finding the normalized eigenvectors with norms equal to unity, an alternative method by using the built-in eigenvalue problem in any numerical package is proposed. A continuous variation of mode patterns for all kind of wave types in the desired frequency range will be rapidly obtained, and the physical meanings of wave propagation can also be shown more clearly and straightforward [8].
The coordinate system which {
Coordinates and displacement orientation.
The equations of motion for the free vibration of a cylindrical shell can be written and shown as below
where
where
In order to transfer the expression from time and displacement domain into frequency and wavenumber domain, the following spectral representation can be further to use [7]
The dimensionless spectral listed in Eq.(1) can becomes as following
where Ω=(
The relationship between s and Ω is so called dispersion relations of cylindrical shell. When a value of dimensionless angular frequency, Ω, is given, non-trivial solutions for the matrix form in the Eq.(4) exist at certain values of s. Finally, the specified roots can be integrated to the polynomial of s2. All the calculating processes and expressions can be rapidly built up by using any kind of symbolic algebra package, such as Matlab or Mathematica. According the physical characteristics of acoustic propagation, two conditions can further be separated and discussed as following.
When n=0, two independent sets of equations and corresponding characteristic equation can be calculated and shown as below:
For axial and radial displacements:
For the non-trivial solution for {U(s,0,Ω), W(s,0, Ω)}, the determinant of matrix in Eq. (5) should be zero ad obtained as below:
where the coefficients, gi, are the function of n and Ω, and shown as
The solutions of Eq.(5) represent a combination of longitudinally and radially expanding and contracting motions; these are the so-called breathing-modes. The expressions of polynomial s2 is obtained in Eq.(6).
Six characteristic roots are given
where
For the circumferential displacement:
The 2nd equation in Eq. (4) can be separated and shown as:
In the same way, under the non-trivial solution for V(s,0,Ω), the new relation can be rewritten as
Obviously, the solution of Eq.(8) represents a uniformly circumferential motion, which is therefore a torsional-mode. The corresponding characteristic roots may be considered the 4th branch pair and show as following
This torsional waves are known for being non-dispersive, since the wave speed is
where c
Under the non-trivial solution of Eq. (4), a 4th polynomial of s2 can be obtained as
These roots yield eight branches of dispersion curves, which are grouped into four branch pairs:
where g
As mentioned earlier, the solutions regarding the modal patterns (eigenvectors) for all four root branch pairs are important. The conventional method has been frequently used for a long time. In this method, the mode shapes are expressed as the ratios among U, V and W. By returning to Eq.(4) after each of the roots (±s
A new alternative method, which solves for the eigenvectors by using a commercially available linear algebra package, will be introduced. This method is extremely simple and straightforward. The kernel of processing is by using the built-in formulation of eigenvalue and eigenvector problem in the numerical package.
According to the eigenvalue problem, the eigenvectors can be found by solving the following homogeneous equations, and the simple flow can be introduce by following:
Step 1. For a specific frequency, dimensionless frequency, Ω, can be obtained and rewritten as Ω
where the asterisk in superscript of L
Step 2. After slightly rearranging, Eq.(13) can be further modified as
Step 3. If Ωs2 is assumed to be unknown for the time being, Eq.(14) can be rewritten to be a typical linear eigenvalue problem and shown as:
In most cases in Eq. (15), for any given value of s (the analytical root), there are three distinct eigenvalues in Ω2 by using the built-in eigenvector function in the numerical package, but only one of them will be equal to Ωs2. Consequently, the eigenvector solution of Eq.(13) is one of the three eigenvectors of Eq.(15), which corresponding eigenvalue is equal to Ωs2. This is because, among the three eigenvalues, only the one that is equal to Ωs will yield the specific s given here, the other two will yield different values of s and are not what we want. When s is specified, the corresponding eigenvector can further calculated by using the numerical solution package, such as MatLab.
The dispersion relations were calculated using the thickness-radius factor β=0.0058 and the Poisson ratio v=0.3. For n=2, the dispersion relation and corresponding modal patterns of Branch pairs 1 are obtained in Fig. 2.(a)and 2(b). The consistency of modal patterns between the alternative and conventional method can also be demonstrated in Fig. 2.(c). The physical characteristics and variations of cylindrical shell at specific circumferential number can be obtained clearly and straightforward.
a) The dispersion curve for Branch 1 roots; (b) The eigenvectors obtained from the current method expressed as [U V W]’; (c) Comparison between the eigenvectors (of Branch 1 roots) obtained from the conventional and the current methods (n=2).
In order to discuss the characteristics of wave propagation in dispersion relations and modal pattern, the numerical results are calculated by assuming the dimensionless thickness parameter, β, to be equal to 0.0058, and the Poisson ratio equals to 0.3. According to Karczub and Leissa, for the usual range of parameters and n≧1, the roots of Eq.(11) were found to have the four following types:
where b
The dispersion curves for all the roots and the corresponding eigenvectors plotted in the figures shown hereafter are those with positive real parts only. In the case of Type 2 roots which have no real part, only the positive imaginary part will be plotted. The roots with negative real parts are not plotted; their eigenvectors are basically the same as those of positive real roots, except there will be sign changes in the longitudinal components if the signs of the radial and circumferential components are chosen to be the same as those plotted.
The dispersion relations and the associated eigenvectors are shown in Fig. 3 and 4, respectively. For simplicity, the eigenvectors [U\' V\' W\'] are plotted by using the absolute values of the displacement components, i.e., [|U| |V| |W|]\'. The dispersion relations for the longitudinal and bending waves of a plate, and those for shear waves are also shown for reference. It shows that at frequencies much lower than the ring frequency (Ω<<1), Branch 1 represents a non-dispersive longitudinal wave, from which it gradually deviates as Ω approaches the neighborhood of unity; Branch 1’s wave eventually approaches that of a plate bending wave when Ω>>1. These are illustrated by the mode shapes shown in Fig. 4(a). Below ring frequency, see the mode shape in Fig. 4(b), Branch 2 (Branch pair 2 root with positive real part) represents the Type 4 root, but it becomes a Type 1 root after crossing the ring frequency and represents the longitudinal wave above the ring frequency, say Ω>1. Therefore, when Ω>1, the Branch 2 curve takes over the role of representing the longitudinal wave which was previously the role of Branch 1’s curve. Below the ring frequency, Branch 3 (Branch pair 3 root with positive real part) represents the Type 3 root, but above the ring frequency, it represents the Type 2 root. The modal components of the Branch 3 roots are predominantly radial (see Fig. 4(c)). Branch 4 (Branch pair 4 root with positive real part) represents the characteristic roots of Eq. (13), which therefore represents the non-dispersive torsional wave and has nearly the same dispersion relation as shear waves. The mode shape of torsional mode is not shown; it is simply [U V W]’=[0\n\t\t\t\t\t1 0]’.
The dispersion relations for n=0 circular harmonic
The normalized eigenvectors of n=0 circular harmonic (no V component)
Fig. 5 and 6 show the dispersion relations and the associated eigenvectors, respectively, for the n=1 circular harmonics. The equivalent curves for Bernoulli beam (labeled as “Berbeam”), Timoshenko beam (labeled as “Timbeam”), and the longitudinal and shear wave curves of a plate are also shown for reference. Branch 1 has only real roots (Type 1 roots); the dispersion curve representing beam bending (when Ω<0.5) agrees well with that of the Timoshenko beam. The Bernoulli beam, on the other hand, agrees with the shell and Timoshenko beam curves only up to Ω=0.03; its applicable frequency range is therefore limited. At frequencies above Ω=0.5, the originally beam-like bending wave gradually transitions to a plate-like bending wave with a predominantly radial modal displacement.
The dispersion relations for n=1 circular harmonic
Below Ω=0.5, the Branch 2 curve is purely imaginary, and represents the Type 2 root. When Ω>0.5, the roots become positive real (Type 1) and gradually approach the shear wave curve, with a predominantly circumferential displacement component when Ω>1.5 (see Fig. 6(b)). Obviously, this branch has a cut off frequency which is about Ω=0.5.
Below the ring frequency, the Branch 3 curve indicates a Type 3 root with a slow and exponentially diminishing wave. When Ω>1.4 it becomes a Type 1 root, and the Branch 3 curve approaches that of longitudinal wave (see Fig. 5 and 6(c)) at further higher frequencies. The propagating waves in Branch 3 thus has a cutoff frequency of about Ω=1.4. The Branch 4 curve indicates a Type 4 root below the ring frequency, but above ring frequency it has only the imaginary part (Type 2) and represents the transversal near field distortion (see Fig. 6(d)).
The normalized eigenvectors of n=1 circular harmonic.
The dispersion relations for the n=2 circular harmonic case are shown in Fig. 7. At very low frequencies, say Ω<0.016, the roots in Branch 1 are the Type 3 roots. At higher frequencies, the roots become Type 1 and represent propagating waves (which thus have a cut off frequency at Ω≒0.016). Fig. 8(a) shows that below ring frequency, Ω<1, the modal patterns of Branch 1 roots have three well-coupled displacement components, although the predominant component is radial. When Ω>1, the modal displacement becomes predominantly radial and the propagating speed asymptotically approaches that of a plate.
The dispersion relations for n=2 circular harmonic
The normalized eigenvectors of n=2 circular harmonic
At very low frequencies, say Ω<0.016, the roots in Branch 2 are the Type 4 roots. Between say Ω=0.016 and the ring frequency, the roots of this branch are Type 2; they represent the near field distortion of the three coupled components. When Ω >1.15, they become Type 1 propagating waves with the wave speeds that asymptotically approach that of the shear wave, and their modal patterns reflect predominantly circumferential shear motions. The curve of Branch 3 indicates a Type 3 root when 0<Ω<1.15, a Type 2 root when 1.15<Ω<2.2, and a Type 1 root when Ω>2.2. This is a propagating wave with a high cutoff frequency (at Ω≒2.2), and at higher frequencies its wave speed approaches asymptotically to that of the longitudinal wave. Branch 4 represents the Type 4 root when 0<Ω≦1.15, which, however, turns into Type 2 and represents the near field distortion when Ω>1.15.
The classic problem of the dispersion of waves in a cylindrical shell had been re-examined with analytical solutions obtained by using a symbolic algebra package. The previous work by Karczub did not include the analytical solutions for the dispersion relations of axisymmetric waves (of circular harmonic order n=0). An axisymmetric wave contains both longitudinal and transverse components; the former are important in the transmission of longitudinal vibration, and the latter are particularly important in acoustics due to their higher acoustic radiation efficiency than the corresponding waves of higher circular harmonic orders (n>0). In this way, a complete set of analytical solutions which based on Flügge shell theory is now available for all orders of circular harmonics, n=0, 1, 2, …, ∞.
A considerable effort has been expended on the solutions for the modal patterns of all propagating and non-propagating modes, because a complete set of properly normalized eigenvectors is crucial to the solution of the vibration problem of a finite shell under various admissible boundary conditions. The eigenvectors obtained by the conventional method are not conveniently normalized as are those commonly used in mathematical physics. A new alternative method to find normalized eigenvectors with norms equal to unity has been proposed and discussed.
Use of analytical solutions has demonstrated the capability of a straightforward continuous tracking on the frequency-dependent changes of the root types and of the corresponding modal patterns for each branch of the dispersion curves associated with a particular analytical root. Therefore, a parallel display of the dispersion curves and of the associated modal patterns used in this paper has provided more insight about the wave phenomena in a cylindrical shell.
The coefficients in Eq. (11) are listed as follows:
where
The coefficients in Eq. (12) are given in the following.
where
and
Recombinant DNA technology involves genetic engineering; cutting DNA molecules from distinct biological species and then ligating them to a vector for expression [1, 2]. The technology helps to express the desired protein in large quantity rather than extracting from bulk amounts of tissues and animal fluids. Proteins are synthesized and modified depending on their functions in an organism. As an initial step, DNA encodes protein through transcription from mRNA synthesis. Then, mRNA is converted into protein. Transcription and translation occur simultaneously in prokaryotic organisms. The conversion of mRNA to protein begins before the synthesis of the mature mRNA transcript [3]. Protein expression involves the synthesis, modification, and regulation of a particular protein in a living organism. However, bacterial systems lack human protein modifications but overexpress recombinant proteins in bulk amounts [4]. Recombinant protein expression is useful to understand the structure and function of proteins. A network of protein complex functions can be distinguished by the characterization of individual proteins function as well as interactions through recombinant protein techniques. Protein–protein and protein-ligand interactions may be highlighted by expressing interacting domains and by introducing key mutations to reveal key domains and residues, respectively. Considering the size and complexity of proteins, protein production is very efficient with vector templates using K12 bacterial systems [2, 4].
Recombinant protein production in bacterial systems is fast, easy, and highly efficient [5]. The general strategy for recombinant protein production involves transforming the cell with a DNA vector containing the open reading frame of the gene, then after subcloning to the expression system, the protein is induced in the cells. After incubating the induced cells, harvested cells are lysed for further separation. The selection of the purification system depends on the type of protein, the affinity tag of the plasmid, the isoelectric point (pI) of the protein of interest, the molecular mass of the protein, the targeted yield, and the degree of functional activity. The lysed cells are purified through column chromatography with a proper resin(s) and a convenient buffer system [6, 7]. But in practice, several steps may cause problems. These include inadequate growth of the selected bacterial host cell, the formation of inclusion bodies, protein aggregation, structural alteration, recombinant protein nonspecific interaction with cellular proteins, problems in colon systems used in purification [6, 8]. Further, as eukaryotic proteins expressed in E. coli may not perform post-translation modifications of other organism proteins, loss of function is observed. In addition, some of the proteins expressed are exposed to hard-denaturing agents or may cause collapses in structure and this often results in insolubility problems. Overexpression of recombinant proteins in bacterial systems leads to the formation of inclusion bodies. Re-folding of these proteins into their bioactive forms is cumbersome and requires a variety of agents and processes [1, 9].
First of all, the choice of the host cells to produce intact protein in the synthesis mechanism forms the mainline of the whole system. Microorganisms used in recombinant protein expression systems include bacteria and yeast. Each host has strengths and weaknesses. The organism to be selected varies depending on the particular protein, working conditions, desired yield. For example, if the desired protein has post-translational modifications, choosing a prokaryotic expression system would not be proper [10]. BL21 (DE3) and its derivatives are by far the most commonly used strains for recombinant protein synthesis. In addition, its genetics are characterized in more detail than any other microorganisms. Recent studies suggest that BL21 (DE3) gene-level research made this bacterium more important for the production of heterologous proteins. This host cell provides maximum efficiency in protein expression through inexpensive substrates, capable of rapid and high-yield growth. A modified form includes a pLysS plasmid that encodes T7 lysozyme. This lowers the background protein expression of recombinant protein but does not perturb IPTG induction. The plasmid is especially useful in toxic cases and provides an option for protein over-expression. Yeast is an alternative recombinant protein production host and provides eukaryotic post-translational modifications with high yield. Yeast growth temperature (30°C) is lower than that of bacteria (37°C) but the growth rate is much slower. Further, the transformation of plasmids to yeast is relatively difficult and the selection of transformed cells and growth conditions require special conditions [10, 11].
The expression plasmids consist of the replication origin, promoter, and multiple cloning sites. The most important issue to consider when choosing an appropriate vector is the copy number property. Because the number of copies is controlled by the replication. It is not always true to assume that the high amount of plasmid is proportional to the yield of recombinant protein expression. Because the high copy is inversely proportional to the rate of bacterial growth. In addition, this condition creates plasmid instability and creates a metabolic load. As a result protein production yield decreases [12, 13].
Prokaryotes have to adapt to the environment by responding quickly to environmental changes. E. coli cells cannot use lactose directly as a source of carbon. But they use glucose, a component of lactose. For the bacterial cell to metabolize lactose, it is necessary to take lactose into the cell and break it down into a glucose monomer. For this, it is necessary to synthesize three different enzymes in the cell [6, 14]. As with E. coli, bacteria combine genes related to the same metabolic pathways to form clusters called operons. Transcription of the genes that make up the operon start from a single promoter. The resulting transcription product consists of an mRNA molecule containing information from multiple genes. Preserved DNA sequences in the promoter region help connect the enzyme to the DNA molecule. Induction is difficult in the presence of easily metabolized carbon sources. If lactose and glucose are present in the environment, expression from the lac promoter is not fully induced until all glucose is used up. In the absence of glucose, the promoter expresses the three enzymes to break down the lactose to obtain glucose. This property is used to induce prokaryotic expression vectors through IPTG (isopropyl 1-thio-β-d-galactopyranoside); a lactose analog that binds lac repressor [14, 15]. In the commercial vectors, IPTG starts the transcription of the lac operon and eventually induces protein expression where the gene of interest is controlled by the
A resistance marker is added to the plasmid to prevent the growth of cells that do not carry plasmids. This can be achieved by using a selection marker. For example, ampicillin resistance is conferred by the
The addition of affinity tags to the plasmid (such as His Tag, glutathione-S-transferase, and cellulose-binding domain) is employed to separate a particular protein from the heterogeneous protein mixture during purification, forming disulfide bonds, increasing the solubility of the recombinant proteins and transferring them to the periplasm region. Affinity tags have a great role in separating the desired protein from cell lysate in recombinant protein purification. Affinity tags are divided into small peptide tags (amino acids) and large polypeptide tags (fusion partners) [17]. Small peptide tags are less likely to interfere when fused to the protein. In some cases, this may have negative consequences on the tertiary conformation and biological activity of the fused chimeric protein. Vectors are available that allow tags to be placed optionally at the N-terminal or C-terminal end. It is more advantageous to position a signal peptide at the N-terminal end for better secretion of the recombinant protein. At this point, it is important to know which end of the protein is embedded in the folding pattern by examining the three-dimensional structure of a particular protein, and it is necessary to place the label on the solvent-exposed end. Examples of small peptide tags are poly-His, c-Myc, and FLAG [18]. His-tagged proteins can be purified by affinity chromatography in resins containing positively charged metal ion nickel. In addition, at the end of purification, with commercial antibodies, labeled recombinant protein can be detected by western blot [17, 18, 19, 20, 21]. On the other hand, it increases the solubility of the recombinant protein produced by the addition of a non-peptide fusion partner (large polypeptide label). The most commonly used fusion labels include Thioredoxin (Trx), Ubiquitin, SUMO, Maltose binding protein (MBP), Glutathione S-transferase (GST) [17, 22]. The reason why fusion partners show properties that increase the solubility of the protein is still not fully explained. Though, MBP label has been shown to carry a small chaperone activity. The GST label has been shown to have the weakest solubility-enhancing effect among fusion partners. Trx has the most solubility-enhancing properties, but due to its size, it may cause adverse effects. In recent years, studies have shown that “Calcium-Binding Protein Fh8” tag derived from a parasite called “Fasciola hepatica” recombinantly added to proteins increases protein solubility [6, 17, 20, 21]. Studies are underway for better solubility enhancing effect of recombinant protein tags.
Even if the effective parameters are provided in the production of recombinant protein, it may not be determined exactly whether the desired protein will be eluted excessively and in active soluble form. Therefore, there are additional strategies for optimizing protein expression [7].
If the desired protein cannot be detected using sensitive techniques or is detected at a low expression rate, the problem is usually caused by a toxic effect of the heterologous protein in the cell. As a result of protein toxicity in the host cell, cells cannot proliferate at a sufficient level and show a low growth rate [7, 23]. The first measure to solve this problem should be followed before proliferating cells are induced. If the growth rate of the recombinant cell is slower than that of the strain with empty vectors, it is related to either gene toxicity or the basal expression of toxic mRNA and protein. Control of basal production is associated with the operon system.
Luria Bertani (LB), the most commonly used growth medium environment for E. coli culture, is an ideal environment for high-nutrient cell growth. When recombinant protein production cannot be replicated with the recommended mechanisms, production efficiency can be increased by increasing the volume of the targeted protein. A successful result can be achieved with adequate ventilation with rigorous shaking of the growth medium. Although LB has a high protein content, cell proliferation is partially reduced. This is due to the low carbohydrate content of LB. As a solution to this situation, increasing peptone and yeast extract provides higher cell proliferation with the addition of MgSO4, which contributes to the sonic intensity of the environment. In addition, the amount of acid released as a result of increased glucose metabolism over time exceeds the buffering capacity of LB. In case of acidification of the growth medium, 50 mM phosphate salts can be added to the environment and buffered [7, 11]. In the broth culture, as the number of cells per unit media increases, oxygen limitation occurs and changes the metabolic capacity of the cell. This prevents optimal growth and the easiest way to increase the amount of oxygen in the growth medium is to increase the speed of the shaking containers. The optimum shaking speed range is 300–400 rpm. Several anti-foaming agents can be added to the broth culture to prevent the negative effect of the foams formed by strong shaking on oxygen circulation [24].
The inclusion bodies formed in E. coli are denatured protein molecules that do not display biological activity. Dissolving, refolding, and purification protocols should be applied, respectively, to make inclusion objects functionally active and soluble. In the transfer of a foreign gene to E. coli, control of gene expression is lost. The nascent polypeptide expression depends on several factors such as osmosis, folding pattern, and pH. If expression increases, the number of unspecific hydrophobic interactions in the polypeptide chain increases. This causes instability and clustering in poly peptization. The resulting protein aggregation is called “inclusion bodies.” The main reason for the formation of clustering is due to the deterioration of the balance between protein aggregation and protein resolution [1, 25]. Therefore, a soluble recombinant protein can be obtained through strategies that eliminate the factors causing the formation of inclusion bodies. As mentioned in the “Affinity tags” section, one way to prevent the solubility problem that may occur in the expressed protein is; combining the desired protein with a fusion partner (large polypeptide tag) that acts as a solubilizer [17].
To obtain the biologically active three-dimensional structure of recombinant proteins, it is important to establish the right disulfide bonds. The formation of improper disulfide bonds causes the protein to fold incorrectly and the formation of inclusion bodies. Disulfide change reactions catalyzed by many enzymes in the Dsb family, where cysteine oxidation occurs in E. coli periplasm, form disulfide bonds in the polypeptide chain [26]. In the cytoplasm, the formation of disulfide bonds is rare because the remnants of cysteine are catalytic regions for many enzymes in the cytoplasm. The wrong disulfide bonds in these regions can cause protein inactivation, clustering, and incorrect folding. However, some strains of E. coli have conditions that trigger the formation of a disulfide bond [5].
Molecular chaperones form the heart of protein synthesis and help nascent polypeptides fold into their active structures. Some specific types of chaperones, such as ClpB, can cleave unfolded polypeptides contained in inclusion bodies. However, high levels of recombinant protein production may result in increased molecular traffic in the cytoplasm, resulting in uncontrolled protein folding control. One strategy used to solve this problem is to arrest protein expression by removing the inducer after a centrifugation step and adding a fresh medium containing chloramphenicol, the protein expression inhibitor. Thus, it allows the recruitment of molecular chaperones to enable the folding of newly synthesized recombinant polypeptides [27, 28]. One of the systems used commercially for protein folding is chaperone plasmids. This system consists of plasmids that allow overexpression of different chaperones or their combinations. Examples of these are GroES-GroEL, DNAK/DNAJ/GrpE [27]. When proteins are released from inclusion bodies, denatured with urea, and subsequently folded
Slowing the production rate of the recombinant protein reduces cellular protein concentration and protein trafficking, allowing the synthesized polypeptides to fold more smoothly. The most common method of reducing the rate of protein synthesis is to lower the incubation temperature [29]. Decreased temperature prevents the formation of aggregation due to its reduction of hydrophobic interactions. Recombinant protein synthesis occurs in the temperature range of 15–25°C. However, when working at the lower temperature range, this causes slower growth and therefore lower protein synthesis. This obstacle can be overcome with commercial products. The ArticExpress™ (Agilent Technologies) competent cells improve recombinant protein expression at low temperatures through co-expressing ortholog genes of E. coli GroEL and GroES from Oleispira Antarctica, namely Cpn60 and co-chaperone Cpn10. These chaperones work together to fold a substrate protein, and usually carry re-folding activity at 4–12°C temperature range, increasing recombinant protein yield and solubility at lower temperatures [30].
Selection of the purification methods generally uses distinct characteristics of the proteins. The distinct properties of recombinant proteins may include chemical, biological, and physical features due to differences in spatial structure and amino acid sequences. Usually, to benefit from these differences, multiple steps are required in the optimal purification process but it should be noted that each step may cause loss of product stability and/or yield, therefore the lowest number of steps are recommended overall for maximum yield. So, method selection determines the ratio of better yield to the better-purified product. Key factors that can affect the purification selection steps include the solubility of the lysate, sample size, and physicochemical properties of the target protein. The first step for purification is to analyze the protein characteristics and match them with literature reports-protocols. For example, a useful parameter in the purification process is amino acid composition. pKa and pI values can be calculated using the amino acid composition. Determination of the values helps to select column type, buffer, pH, or resin type. Once optimization of the purification is established, the method may be employed for a protein with similar sequences or motifs at least for orthologs or isoforms [31]. The characteristic features of proteins that are mainly used for purification type selection are solubility, size, charge, and specific binding affinity [32]. By using these properties, numerous techniques may be employed in protein purification. Solubility parameter can be used with “salting out” through the knowledge of proteins mostly being less soluble in high salt concentrations. And hence, this strategy can be used to separate the protein of interest. Further, dialysis can be used after salting out to remove the salt molecules [33]. Another technique that uses size difference is gel-filtration or size exclusion chromatography (SEC). A column with porous beads resin is used for this and as the sample goes through the column, beads help to separate molecules. The beads are usually 0.1 mm in diameter, so bigger molecules cannot permeate the pores but small molecules penetrate into the pores and are trapped there for a while until the molecules exit again and return to solvent. This action retards small molecules but bigger molecules travel rapidly through a void volume with buffer flow. Small molecules shielding and bigger molecules faster flow separate molecules from each other in fractions depending on their sizes. And as the molecules exit the column, bigger molecules elute first and then, smaller molecules come after.
If the net charge is criteria to be used as a separating feature, ion-exchange chromatography can be used. If the target protein is positively charged as a cationic protein, then, a negatively charged carboxymethyl-cellulose (CM-cellulose) pre-packed column/resin can be used. But if the protein is negatively charged as in anionic proteins, then positively charged diethylaminoethyl-cellulose (DEAE-cellulose) pre-packed columns/resin can be used [33]. It is also known that proteins can have high affinities for certain chemical groups. Affinity chromatography can use this feature to purify proteins and its effect is the best on proteins with affinities to highly specific molecules.
Distinct separation techniques, that is, ion exchange and gel filtration may be employed at high-pressure liquid chromatography (HPLC) with proper column selection. This technique differs from the others because the applied pressure is significantly higher and it does not rely on gravity for sample flow. However, high-pressure limits the purification of higher molecular weight proteins as the pressure denatures protein structure. For higher-molecular-weight proteins FPLC (fast-pressure liquid chromatography) is preferred to prevent pressure-dependent denaturation. FPLC is the preferred technique for protein chemists since any target protein can be separated from cellular lysate readily. And the technique provides a wide range of column options. The flexibility of this technique provides the purification of stable proteins with a high yield. Lower pressure provides advantages as well. Clogging due to lysate content and backpressure problems are less likely encountered compared to that of HPLC. The techniques may also be used with tagged proteins. Histidine tag is one of the most common ones that are used with recombinant proteins and it has a high affinity of metal ions like Ni2+ [17]. To screen if the purification steps are working, gel electrophoresis can be used. In-gel electrophoresis, proteins are separated by their mass as they go through the gel and the smaller ones move faster. As they get separated by their masses, proteins can be visualized in the gel and the gel show protein of interest among others. Another feature to separate proteins is their isoelectric point. This point represents the pH level where the protein has zero net charges. The technique that uses this property to separate proteins is called isoelectric focusing. When proteins go through a pH gradient gel, they will stop at the point where they have no net charge and get separated from the proteins with distinct isoelectric points. To get more specific results, isoelectric focusing can be coupled with SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) in a technique called two-dimensional electrophoresis. In this technique, first isoelectric focusing is done as the sample goes through the gel horizontally and after proteins stop at their respective pH levels, vertical electrophoresis starts. So, the sample is separated according to the isoelectric points horizontally and by their masses vertically. The two-dimensional separation technique is employed to distinguish differences in two different states. Actually, distinct spots may be characterized by MALDI-TOF mass spectrometry.
The structure and function of a protein are essential to characterize the linkage associated between common motifs and biochemical activity [34]. These features are normally determined by NMR or X-ray crystallography techniques [35]. NMR determines dynamic structure but the technique is limited to protein molecular mass. However, an instant picture of the protein structure with a relatively higher mass can be taken by X-ray crystallography. After elucidation of information from these structure determination techniques, scientists concluded that similar sequences show similar structural patterns [36]. Then, different databases which can display protein 2 and 3-dimensional structures have been developed. Determination of protein structure is important for not only understanding the function but also important for protein experiments, such as protein purification. Proteins have alpha helixes, turns, and beta sheets as secondary structures. Beta sheet structures and the outer surface of alpha helixes of proteins can accumulate within the cellular medium or stick to each other and other proteins during aggregation.
Protein structure can be determined by bioinformatic tools such as Swiss Modeling or I-Tasser. Swiss model and I-Tasser are Protein Data Bank (PDB) dependent protein homology modeling databases. They use the known templates from PDB. Swiss Modeling is quicker than I-Tasser, however, I-Tasser produces better and more stable results. Swiss Modeling uses known sequences on the internet and generates data by comparing known structures. Both Swiss Modeling and I-Tasser have the advantage of understanding the main structure of the protein. Protein structure screening is a key factor to understanding interactions of proteins within themselves and with the environment. 3D modeled protein structures can be screened with commercially available tools like YASARA and Discovery Studio Tool. In these tools, not only protein structure is screened, but also domains of proteins can be separated, deleted and water molecules can be removed. UCSF Chimera and Autodock Tools also can be used for screening. After modeling, structures may be downloaded as PDB format and can be visualized by several programs: Discovery Studio, Autodock Vina, UCSF Chimera, etc. Interactions within the protein and secondary-tertiary structures can be obtained from these tools. All these tools work with a PDB file. There are other tools for protein structure determination and can be found at https://www.click2drug.org/. This website provides database links for distinct applications.
Protein databases play a crucial role in bioinformatics and help to find information related to their research. In this way, all biological information becomes accessible through data mining tools saving time and resources. The first step in the study of a new protein is searching databases. Without the prior knowledge from such searches, previously known protein information could be missed, or an experiment could be repeated unnecessarily. There are hundreds of useful databases that can be used in protein research. However, in this study, the pI and 3D structure of the peptide were obtained using the Swiss database and Expasy Database. The purpose of the Swiss Database is to make protein structure modeling accessible. Therefore, with this database, the 3D shape of the protein provided us with information to understand the structure of the peptide and its interactions. Additionally, Expasy Database provides information about proteomics, post-translational modification prediction, primary, secondary, and tertiary structure analysis, sequence alignment, and pI of the protein [37]. The pI of the peptide provides the pH range where that peptide has a negative charge and prepares the buffer solution accordingly. Consequently, databases have key roles in biological research, and enormous data for protein structures, functions, and sequences can be generated by these available databases. These data offer essential information about our protein research as well. Figure 1 provides the predicted structure for our research. SB#14 model indicates that the protein is formed from β-sheet structures.
Predicted structure of Sb#14. Sb#14 is a recombinant synthetic monoclonal antibody 14 used to detect spike protein of COVID-19 and used for immunodetection of the virus. SB#14 is modeled by the Swiss model to design purification steps for the TUSEB project.
Protein solubility is one of the most important protein properties and it can be defined as the protein concentration in a saturated solution that is in equilibrium with a solid phase [38]. Not only some extrinsic factors, including pH, ionic strength, temperature, and some solvent additives, can affect the protein solubility but also several intrinsic factors influence protein solubility. Moreover, the amino acids on the protein surface are the primary intrinsic factors that impact protein solubility [39]. Several studies have revealed the relationship between protein solubility and sequence-derived characteristics. Wilkinson and Harrison et al. provided a simple approach for predicting protein solubility from the sequence, which was further refined by Davis et al. [40]. The average charge, which is derived by the relative quantities of Asp, Glu, Lys, and Arg residues, and the concentration of turn-forming residues are the two parameters used in their solubility model (Asn, Gly, Pro, and Ser). In addition, Christendat et al. have demonstrated that insoluble proteins had more hydrophobic stretches (more than 20 amino acids), less glutamine (Q 4%), fewer negatively charged residues (DE 17%), and a higher percentage of aromatic amino acids (FYW >7.5%) than soluble proteins [41]. The affinity tag (His/GST) in recombinant protein purified by affinity chromatography allows the protein to be purified. However, affinity tag has been observed to alter the biological activity of the protein. Because a minor difference affects protein solubility, the choice of affinity tag at the N- or C-terminus is important when expressing a protein domain. Klock and colleagues investigated a nested collection of 2143N- and C-terminal truncations from 96 targets and found significant variance in both solubility and aggregation processes by changing just a few amino acids in a protein length [42]. Therefore, it is essential to analyze which end of the protein is hidden. Furthermore, if the three-dimensional structure is known, the tag should be kept in a solvent-accessible end. In this way, the solubility of the protein can be increased. Insoluble proteins can aggregate during the expression process. That is why the different parameters should be optimized. On the other hand, during downstream purification steps, protein aggregation can occur. In these cases, developing a suitable and optimized purification procedure for each protein is critical. Sb#14 hydrophobic nature (Figure 1) leads to solubility problems as well as aggregation. The protein sticks to larger proteins and this led to difficulties in purification.
Imidazole is one of the most widely used organic compounds in protein affinity purification processes. It is used as a competitive agent to elute the histidine-tagged proteins. High concentrated imidazole that includes protein samples should be eliminated after eluting from the nickel column by dialysis [43]. In spite of all precautions, his tagged Sb#14 sticks to other proteins and has low solubility, therefore, SEC is used for protein purification rather than affinity purification. As mentioned, SEC separates proteins according to the molecular weight of the molecule. SEC performed with Superdex 75 size-exclusion column 10/30 (GE Healthcare, Princeton, NJ, USA). Moreover, SEC (15 cm length with r, 3 cm column) used in this study separates aggregates readily. Lower molecular weight of Sb#14 provides an advantage in the purification process, recalling that larger proteins elute first. This custom SEC column was unique as the resin resolution is high while the column length is relatively lower. The choice of resin and column size helped to resolve Sb#14 from bacterial lysate in a single purification step. The peptide (MW: 12.468 g/mol, pI: 8.91) is small and prone to aggregate. Therefore, the single-step purification blocks the self-cleavage of protein domains.
The stability of the protein in various buffer compositions and pH levels with and without ligands should be determined. There are some useful websites for fold recognition that can be used to predict the protein fold (PSI-BLAST and SEARCH). Some proteins are misfolded and require the addition of a cofactor, or ligand to restore proper folding and increase stability. For instance, beta-sheets are more prone to form amyloid-like aggregates if there are other binding partners that support protein stabilization and folding [44]. If the protein has a large number of beta-sheets, aggregation may be observed. This can be explained by the tendency of sticking together at Sb#14 and leading to the formation of insoluble aggregates. Tris–HCl buffer is used to stabilize Sb# 14.
To reduce aggregation, reducing agents such as dithiothreitol (DTT) may be used and added to the buffer. DTT is called Cleland’s reagent and is used for protein reduction. However, a high concentration of DTT can reduce the nickel ion in the resin of the column. That is why the determination of the optimal concentration of DTT is essential. β-ME (Beta-mercaptoethanol) cleaves protein disulfide bonds (cystine), and TCEP (Tris phosphine hydrochloride) can also be used as reducing agents, considering longer half-life β-ME. DTT reacts easily with nickel ions whereas β-ME reacts easily with cobalt, copper ions, and other phosphate buffers [44]. A precaution is required to obtain optimal conditions.
Each protein has a pI, where the protein’s net charge is zero. Protein does not migrate at that point, and aggregation occurs [45]. On one hand, acidic proteins are likely to crystallize 0–2.5 pH units above their isoelectric point. On the other hand, basic proteins are more likely to crystallize 1.5–3 pH units below their pI. Hence, different pH values affect the protein’s stability and solubility [46]. The pI of the peptide is important for us to know the pH range where that peptide has a negative charge and to prepare the buffer solution accordingly. That is why the pH of the buffer component is one of the most critical parameters. Sb #14 has a pI value of 8.91. This value set the pH parameter (pH:7.91) of the buffer used in the purification process.
pI represents the pH level of a molecule where the net charge is zero. Amino acid composition of the protein can be used to calculate an estimated value with the help of databases [47]. If the pI is lower than the pH of a solution, protein will have a negative charge but if it is the opposite then the protein will have a positive charge. This feature can be used for purification purposes since it is a specific physicochemical parameter to distinguish between amphoteric molecules [39]. Also, it can be used to understand how solution pH can affect the protein stability in the pH range. So, buffers are used to keep proteins stable. To create an environment for protein to be stable, generally, the buffer is selected to have a pH level around the pI of the protein. If this difference between the pI of protein and pH of the solution gets larger then protein gets a greater net charge too. With this greater net charge, ionic compounds will be able to bind residues [48]. To avoid this unspecific interaction, the buffer’s pH range should be selected accordingly to the protein’s pI. And this knowledge of pH values with their effects on the proteins can be useful in the purification process. In tagged protein purification, affinity chromatography is a commonly used technique. The pH levels also affect this technique since affinity resins have their pH ranges to provide more stable links for not only the ligand and the bead but also for the tag and the ligand. While making decisions about the purification protocol, the affinity resins’ and the tags’ working pH ranges should be kept in mind to create a better environment and more stable interaction. Also choosing affinity resins and tags that have a wider range of pH that they can work may be useful for the purification of proteins.
Proteins are special structures that work with covalent and non-covalent interactions. They have cellular wide roles, including signaling, structural and metabolic processes. Their special structural features and 3D architecture determine their roles and interactions. These forms of proteins are determined by the amino acid sequences [49]. Proteins are not synthesized in their functional form. When their translation process is finished, the primary protein structure is formed. After that, they form alpha helixes and beta sheets by hydrogen bonds. Alpha helixes and beta sheets interact with each other with weak interactions and disulfide bonds and tertiary structure is formed. In some instances, the quaternary structure may be formed when tertiary structures interact and eventually in all cases functional protein forms [55]. However, in some cases, proteins can accumulate and form aggregates which may cause failure in protein purification experiments. Mostly, beta-sheets tend to interact with each other and accumulate. This event can be exampled by amyloid aggregates in Alzheimer’s disease. Recombinant protein aggregates resulted in the prevention of exposure of tags in tagged protein that causes failure in purification. Also, solubility prevents aggregate formation in proteins [50]. Protein aggregation can be prevented by adding salt to the proteins. Salt ions interact with the charged protein surface areas and prevent non-specific interactions, aggregation, and lower protein–protein interaction. However, a precaution is a must when preparing protein for binding experiments. Please note that high ionic concentration blocks ligand/protein binding experiments. As shown in Figure 2, Sb #14 is prone to aggregation and the process may be prevented/decreased through proper conditions.
Ionic strength is important for the separation of proteins from each other that can be aggregated. The strength of ions resists accumulation by preventing protein-protein interaction.
Urea dissolves the aggregated protein solutions. The efficiency of the process is increased by taking the necessary purification steps [1, 51]. Among these processes, protein dissolving and refolding steps constitute the most important steps for optimal protein activity and higher recovery. The protein precipitate is generally separated from other cellular components by low-speed centrifugation after cell lysis. Because protein aggregates are denser than cellular components, the lysate proteins are precipitated by centrifugation and then dissolved using detergents such as urea, guanidine-HCl, high concentrations of chaotropic denaturants, sodium N-lauroyl sarcosine, SDS, N-acetyl trimethyl ammonium chloride [52]. Further, additional reducing agents such as DTT, cysteine, Triton X-100, β-ME are used to dissolve inclusion bodies. These agents retain cysteine residues, minimizing the formation of false and unnatural disulfide bonds in the protein solution. Metal-containing oxidation of cysteine is prevented by using chelating agents such as EDTA in dissolution buffers [44, 52]. By removing the soluble protein content, removing the chaotropic reagents, and diluting them directly into the renaturation buffer, the recombinant proteins are folded back into their native form [44]. Protein collapse is a higher-order reaction while protein folding is a lower-order reaction. Therefore, the aggregation rate is higher than the folding rate. Due to the kinetic competition that occurs, the increase in protein concentration decreases the folding efficiency of the protein. For accurate and efficient folding kinetics, the preferred protein concentration is used in the range of 10–50 μg.ml−1 [1, 53]. As explained in the section of ‘Disulfide bond formation’, recombinant proteins with multiple disulfide bonds in their structure tend to be in a correct folding process in the presence of both oxidizing and reducing agents for the formation of these bonds. The simplest way for oxidation is to oxidize the protein with air in the presence of a metal catalyst. Another common oxidation option is the addition of thiol agents containing compounds such as glutathione, cysteine, cysteamine to the protein mixture. The most commonly used thiol reagents are reduced/oxidized glutathione (GSH-GSSH), cysteine/cystine, DTT/GSSH, cysteamine compounds [1, 36]. There are also low-molecular-weight additives that help refolding process. There are studies on the use of additives such as acetone, DMSO, short-chain alcohols, PEG in the bioactive protein process. In addition, it has been observed that L-arginine/HCl reduces aggregation on protein. The 0.4–1 M arginine used in the studies also increases the protein folding efficiency by reducing the aggregation in the recombinant protein solution. This feature of arginine has been attributed to the interaction of the guanidino structure in its structure with tryptophan residues in proteins [44, 52, 53, 54, 55]. Sb #14 was also treated with DTT but when overexpressed, the protein has solubility problems. The structure is highly prone to aggregation and solubility may be increased upon co-expressing chaperones/Heat Shock Proteins or yeast systems seem proper for preventing aggregation. Yet, this may not solve the problem but mutational studies may provide more soluble and stable structures.
Protein purification depends on several factors: resin type, solvent, ionic strength, pH, protein structural tendency to aggregation, buffer systems, protein structure, ligand if any, column dimension. For each factor, problems may be encountered. To eliminate these problems and decide on protein purification protocol, protein structural properties must be examined initially. Tandem purification steps may also increase the purification yield. However, self-cleavage of certain proteins or oxidation that may distort the protein function leads to problems. Therefore, several distinct protocols may be tested before purifying the targeted protein with high efficiency and functionality. Sb#14 structure mainly consists of β-sheets and overexpressing this petit protein lead aggregation. Solubility is another problem in the cellular milieu as Sb#14 hydrophobic nature interacts with other proteins in the lysate. Therefore, proper solvent selection (phosphate buffer) and adjusting the pH (1 unit lower than pI, 7.91) provide soluble protein. Further, we take advantage of the protein’s lower molecular weight and employed a convenient resin (Superdex 75-separates 3000–70,000 molecular weights, most of the lysate elutes before Sb#14) and custom size column (15 cm length with r: 3 cm-lower pressure yet increase resolution). The purification was performed with high yield by AKTA go FPLC system. Additionally, co-expressing heat shock proteins with this type of protein may help in folding and dissolving aggregates. All these conditions must be tested for individual proteins for optimum purification yield.
Prof. Dr. Yusuf TUTAR acknowledges grant from TUSEB (Project # 8970-220-CV-01) and infrastructure grant from the University of Health Sciences-Turkey (Project #2017-041). Sb#14 is from Addgene (#153522).
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In 1992, Dr. Babinszky obtained a Ph.D. in Animal Nutrition from the University of Wageningen. His main research areas are swine and poultry nutrition. He has authored more than 300 publications (papers, book chapters) and edited four books and fourteen international conference proceedings.",institutionString:"University of Debrecen",institution:{name:"University of Debrecen",country:{name:"Hungary"}}},{id:"201830",title:"Dr.",name:"Fernando",middleName:"Sanchez",surname:"Davila",slug:"fernando-davila",fullName:"Fernando Davila",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/201830/images/5017_n.jpg",biography:"I am a professor at UANL since 1988. My research lines are the development of reproductive techniques in small ruminants. We also conducted research on sexual and social behavior in males.\nI am Mexican and study my professional career as an engineer in agriculture and animal science at UANL. Then take a masters degree in science in Germany (Animal breeding). Take a doctorate in animal science at the UANL.",institutionString:null,institution:{name:"Universidad Autónoma de Nuevo León",country:{name:"Mexico"}}},{id:"309250",title:"Dr.",name:"Miguel",middleName:null,surname:"Quaresma",slug:"miguel-quaresma",fullName:"Miguel Quaresma",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/309250/images/9059_n.jpg",biography:"Miguel Nuno Pinheiro Quaresma was born on May 26, 1974 in Dili, Timor Island. He is married with two children: a boy and a girl, and he is a resident in Vila Real, Portugal. He graduated in Veterinary Medicine in August 1998 and obtained his Ph.D. degree in Veterinary Sciences -Clinical Area in February 2015, both from the University of Trás-os-Montes e Alto Douro. He is currently enrolled in the Alternative Residency of the European College of Animal Reproduction. He works as a Senior Clinician at the Veterinary Teaching Hospital of UTAD (HVUTAD) with a role in clinical activity in the area of livestock and equine species as well as to support teaching and research in related areas. He teaches as an Invited Professor in Reproduction Medicine I and II of the Master\\'s in Veterinary Medicine degree at UTAD. Currently, he holds the position of Chairman of the Portuguese Buiatrics Association. He is a member of the Consultive Group on Production Animals of the OMV. He has 19 publications in indexed international journals (ISIS), as well as over 60 publications and oral presentations in both Portuguese and international journals and congresses.",institutionString:"University of Trás-os-Montes and Alto Douro",institution:{name:"University of Trás-os-Montes and Alto Douro",country:{name:"Portugal"}}},{id:"38652",title:"Prof.",name:"Rita",middleName:null,surname:"Payan-Carreira",slug:"rita-payan-carreira",fullName:"Rita Payan-Carreira",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRiFPQA0/Profile_Picture_1614601496313",biography:"Rita Payan Carreira earned her Veterinary Degree from the Faculty of Veterinary Medicine in Lisbon, Portugal, in 1985. She obtained her Ph.D. in Veterinary Sciences from the University of Trás-os-Montes e Alto Douro, Portugal. After almost 32 years of teaching at the University of Trás-os-Montes and Alto Douro, she recently moved to the University of Évora, Department of Veterinary Medicine, where she teaches in the field of Animal Reproduction and Clinics. Her primary research areas include the molecular markers of the endometrial cycle and the embryo–maternal interaction, including oxidative stress and the reproductive physiology and disorders of sexual development, besides the molecular determinants of male and female fertility. She often supervises students preparing their master's or doctoral theses. She is also a frequent referee for various journals.",institutionString:null,institution:{name:"University of Évora",country:{name:"Portugal"}}},{id:"283019",title:"Dr.",name:"Oudessa",middleName:null,surname:"Kerro Dego",slug:"oudessa-kerro-dego",fullName:"Oudessa Kerro Dego",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/283019/images/system/283019.png",biography:"Dr. Kerro Dego is a veterinary microbiologist with training in veterinary medicine, microbiology, and anatomic pathology. Dr. Kerro Dego is an assistant professor of dairy health in the department of animal science, the University of Tennessee, Institute of Agriculture, Knoxville, Tennessee. He received his D.V.M. (1997), M.S. (2002), and Ph.D. (2008) degrees in Veterinary Medicine, Animal Pathology and Veterinary Microbiology from College of Veterinary Medicine, Addis Ababa University, Ethiopia; College of Veterinary Medicine, Utrecht University, the Netherlands and Western College of Veterinary Medicine, University of Saskatchewan, Canada respectively. He did his Postdoctoral training in microbial pathogenesis (2009 - 2015) in the Department of Animal Science, the University of Tennessee, Institute of Agriculture, Knoxville, Tennessee. Dr. Kerro Dego’s research focuses on the prevention and control of infectious diseases of farm animals, particularly mastitis, improving dairy food safety, and mitigation of antimicrobial resistance. Dr. Kerro Dego has extensive experience in studying the pathogenesis of bacterial infections, identification of virulence factors, and vaccine development and efficacy testing against major bacterial mastitis pathogens. Dr. Kerro Dego conducted numerous controlled experimental and field vaccine efficacy studies, vaccination, and evaluation of immunological responses in several species of animals, including rodents (mice) and large animals (bovine and ovine).",institutionString:"University of Tennessee at Knoxville",institution:{name:"University of Tennessee at Knoxville",country:{name:"United States of America"}}},{id:"251314",title:"Dr.",name:"Juan Carlos",middleName:null,surname:"Gardón",slug:"juan-carlos-gardon",fullName:"Juan Carlos Gardón",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/251314/images/system/251314.jpeg",biography:"Juan Carlos Gardón Poggi received University degree from the Faculty of Agrarian Science in Argentina, in 1983. Also he received Masters Degree and PhD from Córdoba University, Spain. He is currently a Professor at the Catholic University of Valencia San Vicente Mártir, at the Department of Medicine and Animal Surgery. He teaches diverse courses in the field of Animal Reproduction and he is the Director of the Veterinary Farm. He also participates in academic postgraduate activities at the Veterinary Faculty of Murcia University, Spain. His research areas include animal physiology, physiology and biotechnology of reproduction either in males or females, the study of gametes under in vitro conditions and the use of ultrasound as a complement to physiological studies and development of applied biotechnologies. Routinely, he supervises students preparing their doctoral, master thesis or final degree projects.",institutionString:"Catholic University of Valencia San Vicente Mártir, Spain",institution:null},{id:"125292",title:"Dr.",name:"Katy",middleName:null,surname:"Satué Ambrojo",slug:"katy-satue-ambrojo",fullName:"Katy Satué Ambrojo",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/125292/images/system/125292.jpeg",biography:"Katy Satué Ambrojo received her Veterinary Medicine degree, Master degree in Equine Technology and doctorate in Veterinary Medicine from the Faculty of Veterinary, CEU-Cardenal Herrera University in Valencia, Spain. She is a Full Professor at the Department of Medicine and Animal Surgery at the same University. She developed her research activity in the field of Endocrinology, Hematology, Biochemistry and Immunology of horses. She is a scientific reviewer of several international journals : American Journal of Obstetrics and Gynecology, Comparative Clinical Pathology, Veterinary Clinical Pathology, Journal of Equine Veterinary Science, Reproduction in Domestic Animals, Research Veterinary Science, Brazilian Journal of Medical and Biological Research, Livestock Production Science and Theriogenology. Since 2014, she has been the Head of the Clinical Analysis Laboratory of the Hospital Clínico Veterinario from the Faculty of Veterinary, CEU-Cardenal Herrera University.",institutionString:"CEU-Cardenal Herrera University",institution:{name:"CEU Cardinal Herrera University",country:{name:"Spain"}}},{id:"309529",title:"Dr.",name:"Albert",middleName:null,surname:"Rizvanov",slug:"albert-rizvanov",fullName:"Albert Rizvanov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/309529/images/9189_n.jpg",biography:'Albert A. Rizvanov is a Professor and Director of the Center for Precision and Regenerative Medicine at the Institute of Fundamental Medicine and Biology, Kazan Federal University (KFU), Russia. He is the Head of the Center of Excellence “Regenerative Medicine” and Vice-Director of Strategic Academic Unit \\"Translational 7P Medicine\\". Albert completed his Ph.D. at the University of Nevada, Reno, USA and Dr.Sci. at KFU. He is a corresponding member of the Tatarstan Academy of Sciences, Russian Federation. Albert is an author of more than 300 peer-reviewed journal articles and 22 patents. He has supervised 11 Ph.D. and 2 Dr.Sci. dissertations. Albert is the Head of the Dissertation Committee on Biochemistry, Microbiology, and Genetics at KFU.\nORCID https://orcid.org/0000-0002-9427-5739\nWebsite https://kpfu.ru/Albert.Rizvanov?p_lang=2',institutionString:"Kazan Federal University",institution:{name:"Kazan Federal University",country:{name:"Russia"}}},{id:"210551",title:"Dr.",name:"Arbab",middleName:null,surname:"Sikandar",slug:"arbab-sikandar",fullName:"Arbab Sikandar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/210551/images/system/210551.jpg",biography:"Dr. Arbab Sikandar, PhD, M. Phil, DVM was born on April 05, 1981. He is currently working at the College of Veterinary & Animal Sciences as an Assistant Professor. He previously worked as a lecturer at the same University. \nHe is a Member/Secretory of Ethics committee (No. CVAS-9377 dated 18-04-18), Member of the QEC committee CVAS, Jhang (Regr/Gen/69/873, dated 26-10-2017), Member, Board of studies of Department of Basic Sciences (No. CVAS. 2851 Dated. 12-04-13, and No. CVAS, 9024 dated 20/11/17), Member of Academic Committee, CVAS, Jhang (No. CVAS/2004, Dated, 25-08-12), Member of the technical committee (No. CVAS/ 4085, dated 20,03, 2010 till 2016).\n\nDr. Arbab Sikandar contributed in five days hands-on-training on Histopathology at the Department of Pathology, UVAS from 12-16 June 2017. He received a Certificate of appreciation for contributions for Popularization of Science and Technology in the Society on 17-11-15. He was the resource person in the lecture series- ‘scientific writing’ at the Department of Anatomy and Histology, UVAS, Lahore on 29th October 2015. He won a full fellowship as a principal candidate for the year 2015 in the field of Agriculture, EICA, Egypt with ref. to the Notification No. 12(11) ACS/Egypt/2014 from 10 July 2015 to 25th September 2015.; he received a grant of Rs. 55000/- as research incentives from Director, Advanced Studies and Research, UVAS, Lahore upon publications of research papers in IF Journals (DR/215, dated 19-5-2014.. He obtained his PhD by winning a HEC Pakistan indigenous Scholarship, ‘Ph.D. fellowship for 5000 scholars – Phase II’ (2av1-147), 17-6/HEC/HRD/IS-II/12, November 15, 2012. \n\nDr. Sikandar is a member of numerous societies: Registered Veterinary Medical Practitioner (life member) and Registered Veterinary Medical Faculty of Pakistan Veterinary Medical Council. The Registration code of PVMC is RVMP/4298 and RVMF/ 0102.; Life member of the University of Veterinary and Animal Sciences, Lahore, Alumni Association with S# 664, dated: 6-4-12. ; Member 'Vets Care Organization Pakistan” with Reference No. VCO-605-149, dated 05-04-06. :Member 'Vet Crescent” (Society of Animal Health and Production), UVAS, Lahore.",institutionString:"University of Veterinary & Animal Science",institution:{name:"University of Veterinary and Animal Sciences",country:{name:"Pakistan"}}},{id:"311663",title:"Dr.",name:"Prasanna",middleName:null,surname:"Pal",slug:"prasanna-pal",fullName:"Prasanna Pal",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/311663/images/13261_n.jpg",biography:null,institutionString:null,institution:{name:"National Dairy Research Institute",country:{name:"India"}}},{id:"202192",title:"Dr.",name:"Catrin",middleName:null,surname:"Rutland",slug:"catrin-rutland",fullName:"Catrin Rutland",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/202192/images/system/202192.png",biography:"Catrin Rutland is an Associate Professor of Anatomy and Developmental Genetics at the University of Nottingham, UK. She obtained a BSc from the University of Derby, England, a master’s degree from Technische Universität München, Germany, and a Ph.D. from the University of Nottingham. She undertook a post-doctoral research fellowship in the School of Medicine before accepting tenure in Veterinary Medicine and Science. Dr. Rutland also obtained an MMedSci (Medical Education) and a Postgraduate Certificate in Higher Education (PGCHE). She is the author of more than sixty peer-reviewed journal articles, twelve books/book chapters, and more than 100 research abstracts in cardiovascular biology and oncology. She is a board member of the European Association of Veterinary Anatomists, Fellow of the Anatomical Society, and Senior Fellow of the Higher Education Academy. Dr. Rutland has also written popular science books for the public. https://orcid.org/0000-0002-2009-4898. www.nottingham.ac.uk/vet/people/catrin.rutland",institutionString:null,institution:{name:"University of Nottingham",country:{name:"United Kingdom"}}},{id:"283315",title:"Prof.",name:"Samir",middleName:null,surname:"El-Gendy",slug:"samir-el-gendy",fullName:"Samir El-Gendy",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRduYQAS/Profile_Picture_1606215849748",biography:"Samir El-Gendy is a Professor of anatomy and embryology at the faculty of veterinary medicine, Alexandria University, Egypt. Samir obtained his PhD in veterinary science in 2007 from the faculty of veterinary medicine, Alexandria University and has been a professor since 2017. Samir is an author on 24 articles at Scopus and 12 articles within local journals and 2 books/book chapters. His research focuses on applied anatomy, imaging techniques and computed tomography. Samir worked as a member of different local projects on E-learning and he is a board member of the African Association of Veterinary Anatomists and of anatomy societies and as an associated author at local and international journals. Orcid: https://orcid.org/0000-0002-6180-389X",institutionString:null,institution:{name:"Alexandria University",country:{name:"Egypt"}}},{id:"246149",title:"Dr.",name:"Valentina",middleName:null,surname:"Kubale",slug:"valentina-kubale",fullName:"Valentina Kubale",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/246149/images/system/246149.jpg",biography:"Valentina Kubale is Associate Professor of Veterinary Medicine at the Veterinary Faculty, University of Ljubljana, Slovenia. Since graduating from the Veterinary faculty she obtained her PhD in 2007, performed collaboration with the Department of Pharmacology, University of Copenhagen, Denmark. She continued as a post-doctoral fellow at the University of Copenhagen with a Lundbeck foundation fellowship. She is the editor of three books and author/coauthor of 23 articles in peer-reviewed scientific journals, 16 book chapters, and 68 communications at scientific congresses. Since 2008 she has been the Editor Assistant for the Slovenian Veterinary Research journal. She is a member of Slovenian Biochemical Society, The Endocrine Society, European Association of Veterinary Anatomists and Society for Laboratory Animals, where she is board member.",institutionString:"University of Ljubljana",institution:{name:"University of Ljubljana",country:{name:"Slovenia"}}},{id:"258334",title:"Dr.",name:"Carlos Eduardo",middleName:null,surname:"Fonseca-Alves",slug:"carlos-eduardo-fonseca-alves",fullName:"Carlos Eduardo Fonseca-Alves",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/258334/images/system/258334.jpg",biography:"Dr. Fonseca-Alves earned his DVM from Federal University of Goias – UFG in 2008. He completed an internship in small animal internal medicine at UPIS university in 2011, earned his MSc in 2013 and PhD in 2015 both in Veterinary Medicine at Sao Paulo State University – UNESP. Dr. Fonseca-Alves currently serves as an Assistant Professor at Paulista University – UNIP teaching small animal internal medicine.",institutionString:null,institution:{name:"Universidade Paulista",country:{name:"Brazil"}}},{id:"245306",title:"Dr.",name:"María Luz",middleName:null,surname:"Garcia Pardo",slug:"maria-luz-garcia-pardo",fullName:"María Luz Garcia Pardo",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/245306/images/system/245306.png",biography:"María de la Luz García Pardo is an agricultural engineer from Universitat Politècnica de València, Spain. She has a Ph.D. in Animal Genetics. Currently, she is a lecturer at the Agrofood Technology Department of Miguel Hernández University, Spain. Her research is focused on genetics and reproduction in rabbits. The major goal of her research is the genetics of litter size through novel methods such as selection by the environmental sensibility of litter size, with forays into the field of animal welfare by analysing the impact on the susceptibility to diseases and stress of the does. Details of her publications can be found at https://orcid.org/0000-0001-9504-8290.",institutionString:null,institution:{name:"Miguel Hernandez University",country:{name:"Spain"}}},{id:"350704",title:"M.Sc.",name:"Camila",middleName:"Silva Costa",surname:"Ferreira",slug:"camila-ferreira",fullName:"Camila Ferreira",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/350704/images/17280_n.jpg",biography:"Graduated in Veterinary Medicine at the Fluminense Federal University, specialist in Equine Reproduction at the Brazilian Veterinary Institute (IBVET) and Master in Clinical Veterinary Medicine and Animal Reproduction at the Fluminense Federal University. She has experience in analyzing zootechnical indices in dairy cattle and organizing events related to Veterinary Medicine through extension grants. I have experience in the field of diagnostic imaging and animal reproduction in veterinary medicine through monitoring and scientific initiation scholarships. I worked at the Equus Central Reproduction Equine located in Santo Antônio de Jesus – BA in the 2016/2017 breeding season. I am currently a doctoral student with a scholarship from CAPES of the Postgraduate Program in Veterinary Medicine (Pathology and Clinical Sciences) at the Federal Rural University of Rio de Janeiro (UFRRJ) with a research project with an emphasis on equine endometritis.",institutionString:null,institution:null},{id:"41319",title:"Prof.",name:"Lung-Kwang",middleName:null,surname:"Pan",slug:"lung-kwang-pan",fullName:"Lung-Kwang Pan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/41319/images/84_n.jpg",biography:null,institutionString:null,institution:null},{id:"201721",title:"Dr.",name:"Beatrice",middleName:null,surname:"Funiciello",slug:"beatrice-funiciello",fullName:"Beatrice Funiciello",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/201721/images/11089_n.jpg",biography:"Graduated from the University of Milan in 2011, my post-graduate education included CertAVP modules mainly on equines (dermatology and internal medicine) and a few on small animal (dermatology and anaesthesia) at the University of Liverpool. After a general CertAVP (2015) I gained the designated Certificate in Veterinary Dermatology (2017) after taking the synoptic examination and then applied for the RCVS ADvanced Practitioner status. After that, I completed the Postgraduate Diploma in Veterinary Professional Studies at the University of Liverpool (2018). My main area of work is cross-species veterinary dermatology.",institutionString:null,institution:null},{id:"291226",title:"Dr.",name:"Monica",middleName:null,surname:"Cassel",slug:"monica-cassel",fullName:"Monica Cassel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/291226/images/8232_n.jpg",biography:'Degree in Biological Sciences at the Federal University of Mato Grosso with scholarship for Scientific Initiation by FAPEMAT (2008/1) and CNPq (2008/2-2009/2): Project \\"Histological evidence of reproductive activity in lizards of the Manso region, Chapada dos Guimarães, Mato Grosso, Brazil\\". Master\\\'s degree in Ecology and Biodiversity Conservation at Federal University of Mato Grosso with a scholarship by CAPES/REUNI program: Project \\"Reproductive biology of Melanorivulus punctatus\\". PhD\\\'s degree in Science (Cell and Tissue Biology Area) \n at University of Sao Paulo with scholarship granted by FAPESP; Project \\"Development of morphofunctional changes in ovary of Astyanax altiparanae Garutti & Britski, 2000 (Teleostei, Characidae)\\". She has experience in Reproduction of vertebrates and Morphology, with emphasis in Cellular Biology and Histology. She is currently a teacher in the medium / technical level courses at IFMT-Alta Floresta, as well as in the Bachelor\\\'s degree in Animal Science and in the Bachelor\\\'s degree in Business.',institutionString:null,institution:null},{id:"442807",title:"Dr.",name:"Busani",middleName:null,surname:"Moyo",slug:"busani-moyo",fullName:"Busani Moyo",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Gwanda State University",country:{name:"Zimbabwe"}}},{id:"423023",title:"Dr.",name:"Yosra",middleName:null,surname:"Soltan",slug:"yosra-soltan",fullName:"Yosra Soltan",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Alexandria University",country:{name:"Egypt"}}},{id:"349788",title:"Dr.",name:"Florencia Nery",middleName:null,surname:"Sompie",slug:"florencia-nery-sompie",fullName:"Florencia Nery Sompie",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Sam Ratulangi University",country:{name:"Indonesia"}}},{id:"208123",title:"Dr.",name:"Mari-Carmen",middleName:null,surname:"Uribe",slug:"mari-carmen-uribe",fullName:"Mari-Carmen Uribe",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"National Autonomous University of Mexico",country:{name:"Mexico"}}},{id:"345713",title:"Dr.",name:"Csaba",middleName:null,surname:"Szabó",slug:"csaba-szabo",fullName:"Csaba Szabó",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Debrecen",country:{name:"Hungary"}}},{id:"345719",title:"Mrs.",name:"Márta",middleName:null,surname:"Horváth",slug:"marta-horvath",fullName:"Márta Horváth",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Debrecen",country:{name:"Hungary"}}},{id:"420151",title:"Prof.",name:"Novirman",middleName:null,surname:"Jamarun",slug:"novirman-jamarun",fullName:"Novirman Jamarun",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Andalas University",country:{name:"Indonesia"}}}]}},subseries:{item:{id:"11",type:"subseries",title:"Cell Physiology",keywords:"Neurodevelopment and Neurodevelopmental Disease, Free Radicals, Tumor Metastasis, Antioxidants, Essential Fatty Acids, Melatonin, Lipid Peroxidation Products and Aging Physiology",scope:"\r\n\tThe integration of tissues and organs throughout the mammalian body, as well as the expression, structure, and function of molecular and cellular components, is essential for modern physiology. The following concerns will be addressed in this Cell Physiology subject, which will consider all organ systems (e.g., brain, heart, lung, liver; gut, kidney, eye) and their interactions: (1) Neurodevelopment and Neurodevelopmental Disease (2) Free Radicals (3) Tumor Metastasis (4) Antioxidants (5) Essential Fatty Acids (6) Melatonin and (7) Lipid Peroxidation Products and Aging Physiology.
",coverUrl:"https://cdn.intechopen.com/series_topics/covers/11.jpg",hasOnlineFirst:!0,hasPublishedBooks:!0,annualVolume:11407,editor:{id:"133493",title:"Prof.",name:"Angel",middleName:null,surname:"Catala",slug:"angel-catala",fullName:"Angel Catala",profilePictureURL:"https://mts.intechopen.com/storage/users/133493/images/3091_n.jpg",biography:"Prof. Dr. Angel Catalá \r\nShort Biography Angel Catalá was born in Rodeo (San Juan, Argentina). He studied \r\nchemistry at the Universidad Nacional de La Plata, Argentina, where received aPh.D. degree in chemistry (Biological Branch) in 1965. From\r\n1964 to 1974, he worked as Assistant in Biochemistry at the School of MedicineUniversidad Nacional de La Plata, Argentina. From 1974 to 1976, he was a Fellowof the National Institutes of Health (NIH) at the University of Connecticut, Health Center, USA. From 1985 to 2004, he served as a Full Professor oBiochemistry at the Universidad Nacional de La Plata, Argentina. He is Member ofthe National Research Council (CONICET), Argentina, and Argentine Society foBiochemistry and Molecular Biology (SAIB). His laboratory has been interested for manyears in the lipid peroxidation of biological membranes from various tissues and different species. Professor Catalá has directed twelve doctoral theses, publishedover 100 papers in peer reviewed journals, several chapters in books andtwelve edited books. Angel Catalá received awards at the 40th InternationaConference Biochemistry of Lipids 1999: Dijon (France). W inner of the Bimbo PanAmerican Nutrition, Food Science and Technology Award 2006 and 2012, South AmericaHuman Nutrition, Professional Category. 2006 award in pharmacology, Bernardo\r\nHoussay, in recognition of his meritorious works of research. Angel Catalá belongto the Editorial Board of Journal of lipids, International Review of Biophysical ChemistryFrontiers in Membrane Physiology and Biophysics, World Journal oExperimental Medicine and Biochemistry Research International, W orld Journal oBiological Chemistry, Oxidative Medicine and Cellular Longevity, Diabetes and thePancreas, International Journal of Chronic Diseases & Therapy, International Journal oNutrition, Co-Editor of The Open Biology Journal.",institutionString:null,institution:{name:"National University of La Plata",institutionURL:null,country:{name:"Argentina"}}},editorTwo:null,editorThree:null,series:{id:"10",title:"Physiology",doi:"10.5772/intechopen.72796",issn:"2631-8261"},editorialBoard:[{id:"186048",title:"Prof.",name:"Ines",middleName:null,surname:"Drenjančević",slug:"ines-drenjancevic",fullName:"Ines Drenjančević",profilePictureURL:"https://mts.intechopen.com/storage/users/186048/images/5818_n.jpg",institutionString:null,institution:{name:"University of Osijek",institutionURL:null,country:{name:"Croatia"}}},{id:"187859",title:"Prof.",name:"Kusal",middleName:"K.",surname:"Das",slug:"kusal-das",fullName:"Kusal Das",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bSBDeQAO/Profile_Picture_1623411145568",institutionString:"BLDE (Deemed to be University), India",institution:null},{id:"79615",title:"Dr.",name:"Robson",middleName:null,surname:"Faria",slug:"robson-faria",fullName:"Robson Faria",profilePictureURL:"https://mts.intechopen.com/storage/users/79615/images/system/79615.png",institutionString:null,institution:{name:"Oswaldo Cruz Foundation",institutionURL:null,country:{name:"Brazil"}}},{id:"84459",title:"Prof.",name:"Valerie",middleName:null,surname:"Chappe",slug:"valerie-chappe",fullName:"Valerie Chappe",profilePictureURL:"https://mts.intechopen.com/storage/users/84459/images/system/84459.jpg",institutionString:null,institution:{name:"Dalhousie University",institutionURL:null,country:{name:"Canada"}}}]},onlineFirstChapters:{paginationCount:20,paginationItems:[{id:"80964",title:"Upper Airway Expansion in Disabled Children",doi:"10.5772/intechopen.102830",signatures:"David Andrade, Joana Andrade, Maria-João Palha, Cristina Areias, Paula Macedo, Ana Norton, Miguel Palha, Lurdes Morais, Dóris Rocha Ruiz and Sônia Groisman",slug:"upper-airway-expansion-in-disabled-children",totalDownloads:35,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Oral Health Care - An Important Issue of the Modern Society",coverURL:"https://cdn.intechopen.com/books/images_new/10827.jpg",subseries:{id:"1",title:"Oral Health"}}},{id:"80839",title:"Herbs and Oral Health",doi:"10.5772/intechopen.103715",signatures:"Zuhair S. 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