",isbn:"978-1-80355-358-0",printIsbn:"978-1-80355-357-3",pdfIsbn:"978-1-80355-359-7",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!0,isSalesforceBook:!1,hash:"6cfdd7f8f9c18f55c552342a77f63c51",bookSignature:"Prof. Ibrokhim Y. Abdurakhmonov",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11724.jpg",keywords:"Full Genome Sequencing, Mutations and Genetic Variations, Whole-Virus Inactivated and Live-Attenuated, Recombinant Protein-Based, WHO-Protocols, Preclinical and Clinical Trials, New Strain Protection, Availability and Usage, Side Effects, Novel Designs, Country Experiences, Booster-Dose Injection",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"March 17th 2022",dateEndSecondStepPublish:"May 24th 2022",dateEndThirdStepPublish:"July 23rd 2022",dateEndFourthStepPublish:"October 11th 2022",dateEndFifthStepPublish:"December 10th 2022",remainingDaysToSecondStep:"5 days",secondStepPassed:!1,currentStepOfPublishingProcess:2,editedByType:null,kuFlag:!1,biosketch:"A leading biotechnologist, coordinating biotechnology science including COVID-19 vaccine development, laboratory studies, and clinical trials and production in Uzbekistan. Prof. Abdurakhmonov founded (2012) the Center of Genomics and Bioinformatics of Uzbekistan. He received a BS in Biotechnology from the National University, an MS in Plant Breeding from Texas A&M University, USA, a Ph.D. in Molecular Genetics, a Doctor of Science in Genetics, and a full professorship in Molecular Genetics.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"213344",title:"Prof.",name:"Ibrokhim Y.",middleName:null,surname:"Abdurakhmonov",slug:"ibrokhim-y.-abdurakhmonov",fullName:"Ibrokhim Y. Abdurakhmonov",profilePictureURL:"https://mts.intechopen.com/storage/users/213344/images/system/213344.jpg",biography:"Ibrokhim Y. Abdurakhmonov received his B.S. (1997) in biotechnology from the National University, M.S. in plant breeding\n(2001) from Texas A&M University of USA, Ph.D. (2002) in molecular genetics, Doctor of Science (2009) in genetics, and full professorship (2011) in molecular genetics and molecular biotechnology from Academy of Sciences of Uzbekistan. He founded (2012)\nthe Center of Genomics and Bioinformatics of Uzbekistan. He\nreceived the 2010 TWAS prize, and “ICAC Cotton Researcher of the Year 2013” for\nhis outstanding contribution to cotton genomics and biotechnology. He was elected\nas The World Academy of Sciences (TWAS) Fellow (2014) and as a member (2017)\nof the Academy of Sciences of Uzbekistan. He was appointed (2017) as a Minister\nof Innovative Development of Uzbekistan.",institutionString:"Academy of Sciences of Uzbekistan",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"12",totalChapterViews:"0",totalEditedBooks:"12",institution:null}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"16",title:"Medicine",slug:"medicine"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"252211",firstName:"Sara",lastName:"Debeuc",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/252211/images/7239_n.png",email:"sara.d@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. 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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1. Introduction
Direct repair is defined as the elimination of DNA and RNA damage using chemical reversion that does not require a nucleotide template, breakage of the phosphodiester backbone or DNA synthesis. As such, the process of direct repair is completely error-free, granting a major advantage in preservation of genetic information. In mammalian cells, direct repair is utilized to repair specific types of DNA and RNA damage caused by ubiquitous alkylating agents. Only two major types of proteins conduct direct repair in mammalian cells, O6-methylguanine-DNA methyltransferase (MGMT or AGT) and ALKBH family Fe(II)/α-ketoglutarate dioxygenases (FeKGDs). In humans and mice, a single direct repair methyltransferase protein exists, MGMT. In contrast, ALKBH FeKGDs represent a family of nine homologs with conserved active site domains. Although the biochemical function of a number of ALKBH proteins and their biological roles require further investigation, several directly repair alkylation damage in DNA and RNA at base-pairing sites.
2. Direct repair substrates—DNA and RNA alkylation damage
Exposure to alkylating agents is major cause of DNA and RNA damage, generating adducts that can compromise genomic integrity. As a result, repair of alkylation adducts is mediated by a variety of DNA repair pathways, some with overlapping substrate specificity. However, direct DNA repair proteins utilize unique mechanisms to specifically eliminate damage at base-pairing sites. The frequency and site of DNA and RNA damage occurrence is dependent on the source and type of alkylating agent exposure, as discussed in this section.
3. Sources of alkylation damage
Alkylating agents are present environmentally and also generated within the cell via oxidative metabolism. They modify DNA and RNA, forming adducts that disrupt replication and transcription, trigger cell cycle checkpoints, and/or initiate apoptosis. If left unrepaired, some adducts formed by alkylation damage can be cytotoxic and/or mutagenic [1-3].
Environmental alkylating agents fall into two primary groups, nitrosoureas that generate primarily O-alkylations and methanesulfonates that cause mostly N-alkylations [1, 3] (Figure 1). These exogenous alkylating agents are present in air, water, plants and food, in the form of nitrosamines, chloro- and bromomethane gases, myosamines and halocarbons [4]. There are also industrially produced alkylating agents, including various chemotherapeutic agents [5, 6].
Figure 1.
Examples of nitrosourea and methanesulfonate alkylating agents. (A) Nitrosourea, SN1, alkylating agents. Abbrevations are as follows: methylnitrosourea (MNU); ethylnitrosourea (ENU); 1,3-bis (2chloroethyl)-1-nitrosourea (BCNU); N-(2-chloroethyl)-N-cyclohexyl-N-nitrosourea- (CCNU); N-methyl-N-nitro-N-nitrosoguanidine (MNNG); N-ethyl-N-nitro-N-nitrosoguanidine (ENNG). (B) Methanesulfonate, SN2, alkylating agents. Abbrevations are as follows: dimethylsufate (DMS); diethylsulfate (DES); methylmethanesulfonate (MMS); ethylmethanesulfonate (EMS). [14]
Enzymes involved in cellular metabolism are responsible for the majority of endogenous alkylating agent damage. Nitrosating agents are generated, resulting in amine nitrosation, and reactive oxygen species (ROS), which cause lipoperoxidation [7]. Additionally, a family of S-adenosyl methionine (SAM) methyltransferase enzymes is involved in more than 40 metabolic reactions using SAM as a methyl donor to modify nucleic acids, proteins and lipids [8, 9]. Four of those SAM methyltransferase enzymes participate in DNA and RNA modification in mammalian cells. DNMT1, DNMT3A, and DNMT3B catalyze methyl group transfer at the C5 position of cytosine in DNA CpG sequences [10], whereas TRDMT1 (DNMT2) methylates the C5 position of cytosine 38 in aspartic acid tRNA [11].
3.1. Types of alkylating agents
Alkylating agents can be categorized by their method of activation. Some alkylating agents react directly with DNA and do not require any activation, whereas many alkylating agents, including many carcinogens, must undergo metabolic activation by the cytochrome P450 system to generate reactive species capable of modifying DNA [3, 12, 13]. In addition, alkylating agents are electrophilic compounds that possess either one or two reactive groups that can interact with the nucleophilic centers of DNA and RNA bases. Alkylating agents that can only react with one nucleophilic center are mono-functional, whereas bi-functional agents can react with two sites in DNA or RNA [1, 13]. Alkylating agents that are mono-functional primarily transfer alkyl groups to ring nitrogens, while agents that react in a bi-functional manner not only react with ring nitrogens, but can form cyclized DNA bases, by reacting with exocylic nitrogen and oxygen groups [13] (Figure 2). In addition to methylating agents, larger alkylating agents also modify nucleic acids—bi-functional ethylating agents can form exocyclic ethano and etheno adducts at nitrogen and oxygen molecules in all DNA and RNA bases. Additionally, bi-functional alkylating agents can produce DNA inter- and/or intrastrand cross-links [13]. Some alkylating agents also react at phosphate residues to generate phosphotriesters, leading to potential single-strand breaks [13] (Figure 2). Two main pathways, characterized as SN1 or SN2, are defined based on the kinetics of the alkylation reaction, leading to the above mentioned modifications of DNA and RNA bases [2].
Figure 2.
(A) Purple arrows indicate sites in DNA most often methylated by SN1 alkylating agents. Green arrows indicate sites commonly modified by SN2 alkylating agents, orange arrows indicate sites in single-stranded DNA. Blue arrows indicate exocyclic amino groups important in formation of cyclized DNA adducts. The location of the major and minor grooves in DNA are indicated. “R” is the attachment of the base to the deoxyribose and phosphodiester backbone. (B) Modified phosphodiester isoforms in the DNA backbone. SN1 alkylating agents generally form more phosphotriester products than SN2 agents. [2,14]
SN1 agents act via a two step reaction involving a unimolecular nucleophilic substitution with a rate-limiting step that generates an intermediate carbonium ion electrophile that reacts with nucleophilic DNA sites. Thus, the reaction kinetics depend only on the formation of the carbonium ion intermediate (first-order). The triganol planar conformation of the sp2 hybridized carbon generated in the carbocation intermediate permits nucleophilic attack from either side, yielding a racemic mixture of reaction products at chiral centers [13] (Figure 3). Though agents that react via an SN1 mechanism produce both N- and O-alkylations, increased amounts of modified oxygens are generated, compared to agents that react via an SN2 mechanism.
Figure 3.
SN1 and SN2 nucleophilic substitution reactions. (A) Example of an SN1 reaction. SN1 reactions are dependent on formation of a carbonium ion intermediate that rate-limiting. Product chiral centres are a racemic mixture because the intermediate can be attacked by either side. (B) Example of an SN2 reaction. Both reactants are required and there is direct attack by the nuclephile in SN2 reactions. Chirality is maintained since a transition state is formed with the chiral center. [2,14]
In contrast, SN2 reaction mechanisms depend on both the alkylating agent and its target to define the kinetics (second-order). Using a one step reaction where both the electrophile and nucleophile are involved in the transition state, SN2 alkylating agents proceed with direct attack by the nucleophile on an electron deficient center. The nucleophile attacks from the back of the electrophile, forming the carbon-nucleophile bond and breaking the carbon-leaving group bond. Simultaneous backside, nucleophilic attack and leaving group departure cause the incoming group to replace the leaving group. Because a transition state is formed with the chiral center, chirality is maintained, leading to a stereocenter (inversion) configuration [13] (Figure 3). Alkylating agents that react via an SN2 mechanism cause primarily N-alkylations.
3.2. DNA and RNA alkylation damage
Modification sites of DNA bases are the same for all alkylating agents and include all the exocyclic nitrogens and oxygens, as well as ring nitrogens without hydrogen. Though all DNA nucleobase oxygen or nitrogen atoms can be alkylated, the type and frequency of specific damage varies depending on the type of alkylating agent, the structure of the substrate, and the position of the damage site [13] (Table 1). Generally, alkylation damage at nitrogen molecules is less mutagenic than oxygen, though both types of alkylation damage are cytotoxic and genotoxic [14].
Common alkylations generated by exogenous alkylating agents include O6-alkylguanine and O4-alkylthymine adducts, as well as N7-alkylguanine, N3-alkyladenine, N1-alkyladenine, and N3-alkylcytosine [13] (Figure 1). Moreover, the frequency of each adduct type depends on whether the DNA and RNA substrates are single- or double-stranded [13] (Table 1). For instance, nitrogen molecules involved in DNA base-pairing are less vulnerable to alkylation damage than the same base nitrogens in a single-stranded region arising during replication and transcription.
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t
\n\t
Table 1.
% of Total DNA alkylation adduct formation in single- and double-strand DNA. Modifications following SN2 alkylating agent methylmethanesulfonate (MMS) or SN1 alkylating agent treatments methylnitrosourea (MNU) or ethylnitrosourea (ENU). Sites where % alkylation is undetermined are indicated as (--) [13].
4. Direct repair proteins
Numerous cellular mechanisms have evolved to deal with various types of DNA damage and each DNA repair pathway is important to maintain genomic integrity. However, most repair mechanisms require DNA synthesis and therefore an intrinsic risk of causing mutation in executing the repair. In contrast, direct repair proteins, MGMT and ALKBH family proteins employ direct reversal mechanisms that result in complete restoration of DNA bases and are thus error-free mechanisms. Moreover, MGMT, ALKBH2, and ALKBH3 repair endogenous and exogenous DNA and RNA alkylation damage at critical base-pairing sites, facilitating proper replication of genetic information or transcription. This section will discuss each of these direct DNA repair enzymes in detail.
Figure 4.
Major mechanisms of alkylation adduct repair. Direct repair pathways are indicated in green. Base and nucleotide excision repair pathways are indicated in blue [2,14].
4.1. Mechanisms of alkylation repair
Multiple mechanisms are employed to rid the genome of alkyl adducts, thereby preventing detrimental effects within the cell (Figure 4). Mismatch repair (MMR), base excision repair (BER) and nucleotide excision repair (NER) and direct repair (DR) pathways all participate in alkylation damage repair [15-24]. Specifically, BER and NER repair small alkylated base damage including 7-methylguanine (7-meG) and 3-methyladenine (3-meA) DNA adducts [25]. Although BER repairs the majority of small alkylated base damage (methyl and ethyl adducts) the NER system can also remove small, as well as bulky adducts larger than ethylated bases [24, 26]. As an alternative to NER, incomplete BER repair intermediates can be processed by homologous recombination (HR) [27]. However, BER, NER and HR repair pathways generate strand breaks during repair of alkyl adducts and could introduce mutations or rearrangements [28]. On the contrary, DR mechanisms, provided by methyltransferase MGMT and ALKBH homologs, eliminate alkylation damage at DNA base-pairing sites, including O6-methylguanine (O6-meG), 1-methyladenine (1-meA) and 3-methylcytosine (3-meC) and do not require a nucleotide template, result in phosphodiester backbone breakage, nor do they require DNA synthesis.
In mammals, methylguanine DNA methyltransferase (MGMT or AGT), can repair two types of DNA adducts: O6-methylguanine (O6-meG) and O4-methylthymine (O4-meT). O6-meG adducts in DNA are extremely mutagenic [29, 30] and also block DNA polymerase extension, which is generally associated with cytotoxicity [31, 32]. The primary mutations observed when there is a failure to repair O6-meG adducts prior to replication are G:CA:T transitions, whereas a failure to repair O4-meT results primarily in T:AC:G transition mutations [29]. In mammals, elimination of O6-meG by MGMT is preferred over O4-meT, but the respective efficiency of each type of reversion is species dependent [29, 33-37].
Removal of O6-meG and O4-meT modifications are achieved via a one-step methyltransferase reaction, wherein MGMT accepts the alkyl adduct from the modified oxygen molecule, onto an internal residue, directly restoring the DNA base and inactivating the protein [38] (Figure 5). In addition to methyl groups, several other alkyl-adducts can also be transferred from guanine to MGMT, including ethyl-, propyl- butyl-, benzyl- and 2-chloroethyl-. However, the efficiency of the reaction is decreased for alkyl adducts greater than methylated bases [39]. Once modified, the protein is targeted for elimination via the proteasome [40].
4.2.1. Protein structure/active site organization
Alkyltransferase proteins are found in eukaryotic and prokaryotic organisms and have been identified in as many as 100 organisms [41]. Though sequences are not highly conserved between human MGMT and Eubacterial, Archea, and Eukaryotic DNA methyltransferase enzymes, structural domains and active site residues are almost identical [42-46].
Figure 5.
Methylguanine methyltransferase (MGMT) activity. (A) MGMT DNA repair substrates (B) MGMT repair reaction. Transfer of the methyl group (orange) from the damaged DNA base to the internal Cys145 (light green) is a suicide reaction, inactivating MGMT. [14]
In human MGMT, a conserved α/β roll structure, containing a three-stranded, anti-parallel β-sheet, followed by two helices, make up the N-terminus (residues 1-85). The MGMT C-terminus (residues 86-207) contains a short, two-stranded, parallel β-sheet, four α-helices and a 310 helix [42, 47]. Found only in humans, a zinc ion stabilizes the interface between the N- and C-termini, binding Cys5, Cys24, His29 and His85 in a tetrahedral conformation to bridge three strands of the N-terminal β-sheet with the coil preceding the 310 helix in the C-terminus [47].
The conserved active site cysteine motif (-PCHR-) is located in the C-terminus contained within the DNA binding channel, and the helix-turn-helix (HTH) DNA binding motif. Residues Try114-Ala121 form the first helix of the HTH motif and residues Ala127-Gly136 form the second, “recognition” helix, which interacts with DNA. Linked by an Asn-hinge (Asn137) that stabilizes the over-lapping turns by binding Val139, Ille143 and the Cys145 thiol, the -PHCR- active site is located near the “recognition” helix [42, 47, 48].
The active site of human MGMT is composed of at least ten residues that participate in substrate binding, enzyme structure and alkyl transfer. Residues Val155-Gly160 and Met134 generate a hydrophobic cleft in the active site loop, while residues Tyr114, His146, Val148, Ser159, and Glu172 participate in active site coordination and alkyl group transfer to residue Cys145. Not unexpectedly, mutation of residue Cys145 results in elimination of alkyl group transfer, however substrate binding is unaffected [49] (Figure 6).
Figure 6.
Structure of human MGMT (PDBid1QNT). The N-termianl p-sheet and C-terminal 310 helix of the α/β roll structure, conserved in AGT proteins are indicated. In humans, a zinc ion stabilizes interaction of MGMT N-and C-termini [46].
4.2.2. Substrate recognition/repair mechanism
In repair, MGMT is unique in that one molecule is responsible for the removal of one O6-meG or O4-meT adduct. Unlike most enzymes with the capacity to catalyze multiple reactions, MGMT catalyzed reactions are stoichiometric and capable of only a single repair reaction [50]. As a result, removal of O6-meG and O4-meT alkyl adducts is dependent on both MGMT and the substrate concentrations (second-order reaction).
The recognition of guanine and thymine base methylation is accomplished by a highly conserved amino acid structure. The hydrophobic cleft of the active site loop and -PCHR- motif within the binding channel allow MGMT to bind to the minor-groove of DNA using residues Ala126, Ala127, Ala129, Gly131, and Gly132, of the HTH “recognition” helix [51, 52], which is followed by necessary conformational changes to orient the damaged base within the active site.
Identified based on bacterial Ada homology and human MGMT structures, following substrate recognition, the target base is repaired using a base flipping mechanism [53-58]. In the MGMT repair reaction, the damaged base undergoes a residue Tyr114-mediated, sterically enforced 3’ phosphate rotation into the active-site pocket. The hydrophobic cleft formed by the active site loop easily accepts the extra-helical base, causing the DNA minor groove to widen [51]. The arginine finger residue, Arg128, intercalates between the DNA bases and interacts with the unpaired cytosine, via a charged hydrogen bond [55], maintaining an appropriate DNA duplex conformation (Figure 6).
Once bound within the MGMT active site, numerous residues participate in the methyltransferase reaction. A hydrogen bond network, conserved in AGTs, is formed between Glu172, His146, water and Cys145. His146 acts as a water-mediated base that deprotonates Cys145, converting Cys145 to a cystine thiolate anion and generating an imidazolium ion that is stabilized by Glu172 [35, 59]. Residues, Val148 and Cys145 carbonyls accept guanine exocyclic amine hydrogen bonds and nitrogen atoms of residues Tyr114 and Ser159 donate protons to N3 and O6 of O6-meG, respectively. The deprotonated Tyr114 residue abstracts a proton from Lys165, simultaneously transferring the alkyl group from the O6 position of guanine to the thiolate anion of the Cys145 residue [35]. Transfer of the alkyl group generates a thioether, S-alkylcysteine, and results in complete restoration of the guanine base, as well as irreversible inactivation of the methyltransferase enzyme (Figure 5). While many DNA repair proteins have a specific requirement for double-stranded DNA, MGMT can also bind to single-stranded DNA [60].
4.2.3. Gene expression/protein regulation
Removal of O6-meG modifications by MGMT has a major role in cell cycle checkpoint control, proliferation, and differentiation [61]. As a result, MGMT is a house-keeping gene that is expressed in all tissues; though expression varies depending on cell type [62]. MGMT expression in an individual cell or tissue type is dependent on a variety of factors, including numerous types of stimuli and promoter regulator elements. However, the relationship between factors that mediate MGMT expression and the regulation of its function is not well-understood. The lack of understanding regarding the consequences of MGMT regulation is illustrated by the fact that MGMT expression is silenced in some cancers, but expression is up-regulated in others [62, 63].
MGMT is a single gene on chromosome 10q26, spanning approximately 300kb [64]. The gene has five exons, but the first is non-coding [65, 66]. The promoter of MGMT is a non-TATA-box promoter that contains a GC-rich CpG island of 780 bp that includes 97 CpG dinucleotides [67]. CpG islands are commonly associated with promoter regions of constitutively expressed genes, from which transcription is initiated from a single promoter site [68-70]. Additionally, the promoter contains six transcription consensus binding sites (SP1, AP1, and AP2), three upstream and three downstream of the transcription start site, a glucocorticoid-responsive element, and a 3’ enhancer element [62, 67, 69, 71]. Though unmethylated in normal cells, promoter CpG island methylation-induced silencing of MGMT is found in various cancer types and MGMT-deficient cell lines and is one mechanism that regulates MGMT expression [72-76]. However, whether MGMT promoter methylation disables transcription factor binding or contributes to chromatin reorganization remains uncertain [71, 75].
In addition to numerous transcription factor binding sites that surround the MGMT promoter transcription start site, the MGMT promoter CpG islands exhibit a chromatin structure that mediates interaction with transcription factors. The MGMT gene is organized around five or more nucleosomes in a manner that positions 300 bp region of the promoter sequence, which contains known MGMT transcription factor binding sites, so that it does not lie within the nucleosomes, and therefore does not maintain a higher-order chromatin structure [62, 72, 77]. Such nucleosomal positioning facilitates an “open” stretch of DNA that enables constitutive interaction of transcription factors with the promoter.
Methylation of the CpG island surrounding the transcription factor binding sites contributes to lack of transcription factor binding, but could also effect nucleosomal positioning of the MGMT promoter [62, 71], suggested by histone H3 Lys9 (H3K9) di-methylation, exhibited in relationship to MGMT silencing [78, 79]. Further, deacetylation of histones H3 and H4 could also be associated nucleosome organization that is more condensed, resulting in transcription inactivation. Therefore, the chromatin structure of the MGMT promoter, as well as CpG island methylation, mediate transcription factor access to the promoter and are important for MGMT expression.
4.2.4. Protein localization and cell type dependence
Immunofluorescence studies indicate MGMT nuclear localization at discrete nuclear regions [80]. Although a nuclear localization signal (NLS) for MGMT has not been identified, the small size of MGMT, 23 kDa, may not require an active translocation signal to traverse nuclear pores [53]. However, a –PKAAR- sequence within the DNA binding domain of MGMT is necessary for DNA interactions to facilitate nuclear retention [81]. The highest MGMT expression levels are found in the liver, where high levels of endogenous nitrosating agents are present, but MGMT is also expressed at high levels in the lung, kidney and colon. MGMT expression is heterogeneous in the brain and the lowest levels are observed in the pancreas, hematopoietic cells, lymphoid tissues [62, 67, 82-86].
4.2.5. Post-translational modification
Once MGMT has transferred a methyl group to its Cys145 residue, no further reactions are catalyzed, so the protein must be eliminated. The degradation of MGMT is an ubiquitination-dependent process that has been evaluated using inactivation of the protein by O6-BzG, BCNU, or NO-generating agents at position Cys145 [40, 87, 88]. Conformational changes in the protein structure after alkyl group transfer target MGMT for ubiquitination and proteasomal degradation [40, 89]. Two sites within MGMT, Lys125 and Lys178, have been identified as ubiquitination targets in B lymphocyte (NCI-H929) or 293T, and myeloid (MV4-11) cells, respectively. Additionally, examination of potential MGMT modification sites using predictive software also identifies Lys104 as an ubiquitination target. Furthermore, predictions also indicate post-translational modification sites for methylation (Arg128, Arg135), acetylation (Lys8, 125, 178, 193), and sumoylation (Lys75, 205, 18, 107), as well as numerous phosphorylation sites (Ser36, 56, 130, 182, 202, 206, 208; Thr37; Tyr91, 115) [90-93], which all merit further consideration. Notably, phosphorylation of residues Thr10 and Thr11 was also noted in HeLa cells [92], and phosphorylation of Ser201 is observed in B lymphocyte cells (DG75 and GM00130), KGI myeloid cells, and HeLa cervical cancer cells. Importantly, crystallographic data suggests that modification of Ser201 could disrupt interaction with DNA [48, 51, 55].
In mammals, repair of cytosine and adenine base methylation at base-pairing positions is specifically associated with the AlkB family dioxygenase proteins [92, 94-96]. Discovered first in Escherichia coli (E. coli) in 1983 [96] alkylation protein B (AlkB) belongs to a super-family of Fe(II)/α-ketoglutarate-dependent dioxygenases (FeKGDs), with roles in histone demethylation [97-99], proline hydroxylation [95] and in the case of AlkB, the ability to directly remove alkyl adducts generated in DNA residues as a result of exposure to SN2 alkylating agents [94, 100]. Originally predicted to act on 1-methyladenine (1-meA) and 3-methylcytosine (3-meC), bacterial AlkB has been shown to repair a variety of DNA and RNA adducts, including 1-meA, 3-meC, 1-meG, 3-meT, 1-etA, as well as aromatic ethyl, 3-etC, and etheno adducts, 1,N6-ethenoadenine (εA) and 3,N4-ethenocytosine (εC) [94, 100-108] (Figure 7).
Figure 7.
ALKBH protein substrates. (A) DNA methyl adducts repaired by ALKBH proteins. (B) DNA etheno adducts repaired by ALKHB proteins.
Using bioinformatics, nine human ALKBH family enzymes, ALKBH1-8 and FTO, were identified, of which only four have been reported to have DNA repair activity, ALKBH1 – ALKBH3 and FTO [109, 110]. Though all of the ALKBH homologs contain conserved catalytic domain residues, none entirely encompass the enzymatic activity of AlkB [15, 103, 104, 111-114]. Removal of alkyl adducts from DNA is only accomplished by three ALKBH proteins, ALKBH1-3, known to remove 1-meA and 3-meC adducts. However, ALKBH1 is reportedly a mitochondrial protein [115], therefore in the nucleus ALKBH2 and ALKBH3 proteins are employed to remove specific adducts in single- or double-stranded DNA or in RNA [104]. Lesions that are repaired by ALKBH proteins generally interfere with base-pairing and block replication and transcription, triggering cell cycle checkpoints and apoptosis [92, 95, 96, 110, 115]. In E. coli AlkB mutants, as well as in Alkbh2- or Alkbh3-deficient mouse embryonic fibroblasts, cells exhibit increased sensitivity to alkylating agents, particularly the SN2 type, and increased mutant frequency [101, 116-119].
4.3.1. Protein structure/active site organization
Similar to MGMT, the sequences of human ALKBH proteins do not contain a high percentage of sequence homology in regions other than active sites and conserved domains, but do have conserved secondary structures [109, 110, 114, 120-122]. In AlkB family proteins, the catalytic core is composed of three major components, the double-stranded β-helix (DSBH), the nucleotide recognition lid (NRL) and the N-terminal extension (NTE) (Figure 8). The DSBH is comprised of eight β-strands in the C-terminal portion that form two β-sheets to create a central core jelly-roll fold. Within the major and minor β-sheets of the DSBH lie conserved catalytic residues RxxxxxR and HxDxnH, respectively [120, 121, 123]. The HxD dyad is near the amino terminal end and is located in a flexible loop that follows the first strand, stacking with the minor β-sheet. The carboxy-terminal histidine of the conserved HxDxnH residues is associated with the beginning of the sixth strand and together these residues coordinate iron (His171, Asp173 and His236—Alkbh2; His191, Asp193 and His258—Alkbh3) [114, 120, 121, 123, 124]. The histidine and aspartic acid residues (Asp248 and Asp254—ALKBH2; Asp269 and Asp275—ALKBH3), conserved in the DSBH minor β-sheet, coordinate Fe(II), α-ketoglutarate and the DNA or RNA repair substrate within the catalytic core. A conserved Arg residue in the C-terminal β-strand (Arg254—ALKBH2 and Arg275—ALKBH3) sets AlkB family proteins apart from other α-ketoglutarate-dependent dioxygenases within the Fe(II)/α-ketoglutarate dioxygenase superfamily, forming the base of the substrate binding pocket [110, 120, 121, 123].
Figure 8.
Structure of human AlkB homolog DNA repair proteins. Two looped structures (flip1 and flip2) generated by anti-parallel β-sheets create the nucleotide recognition lid (NRL) and are involved in DNA base flipping. (A) Structure of ALKBH2 (PDBid3BTX). ALKBH2 double-strand DNA substrate specificity is facilitated by residues in loops L1 and L2. (B) Structure of ALKBH3 (PDBid2IUW). β -sheets 4 and 5 form the β-hairpin motif in ALKBH3. Part of loop 1, involved in ALKBH substrate specificity, was omitted due to electron density problems. [121]
The N-terminal extension (NTE) and Nucleotide Recognition Lid (NRL) are formed by the β-hairpin motifs that extend from the DSBH jelly-roll, forming a substrate binding groove that covers the active site until bound. Ninety residues are contained within two looped structures, forming “flips” that lie between a single β-sheet and two α-helices in the N-terminal portion of the catalytic core [120, 121]. Secondary structures are of similar size, but possess different characteristics important for substrate specificity and DNA activity. In ALKBH2, the first flip is 20 residues that make up a β-hairpin and short α-helix, creating a hydrophobic binding groove. In contrast, the first flip in ALKBH3 is a β-hairpin made up of 17 residues that form a hydrophilic, positively charged binding groove, more suitable for single-stranded DNA or RNA substrates [15, 120]. The characteristics of the second flip are also unique. Flip two of ALKBH2 spans 24 residues that is made up of three β-sheets, with numerous sites for DNA substrate interaction. The orientation of the three β-sheets, which fold back towards the C-terminal end of the first α-helix, is also unique only to ALKBH2 [114, 121]. However, flip 2 of ALKBH3 is only 12 residues and contains a single β-sheet [114]. The N-terminal regions of each ALKBH homolog are more variable and hypothesized to play roles in sub-cellular sorting and protein-protein interactions [114, 115] (Figure 8).
In addition to the conserved catalytic dioxygenase residues, some human ALKBH proteins also contain additional catalytic residues and domains [104, 109, 110, 113, 125] (Figure 9). Structural analysis of bacterial AlkB and human ALKBH homologs provides insight into substrate preferences and repair capabilities. For instance, ALKBH2 contains three unique motifs that facilitate enhanced activity on double-stranded DNA [121]. A long, flexible β-sheet hairpin loop that contains DNA binding residues Arg198, Gly204 and Lys205, a short loop that contains the RKK motif (Arg241-Lys243) and an aromatic finger residue (Phe102) are used to make contacts with both DNA strands, rotate and take the place of the damaged base in duplex DNA molecules. On the other hand, the number and organization of the catalytic domains in ALKBH3 result in differential manipulation of the DNA backbone, explaining the preference for single-strand substrates. Lack of an aromatic finger residue and RKK motif in ALKBH3, the damaged base is squeezed on either side, forcing it to rotate, and the immediate 5’ and 3’ bases to stack against one another. However, structural analysis of ALKBH3 has identified residue Arg122, specifically the arginine side chain length, as important for double-stranded DNA substrate activity, possibly mimicking the base-flipping and stacking activities of ALKBH2 residue Phe102 [114, 121].
Unfortunately, extensive biochemical analysis or structural studies have not been conducted on ALKBH homologs 4-8. However, it is apparent that differences in the number and organization of catalytic residues, as well as secondary structures play a large role in the diversity of ALKBH family protein substrate specificities and enzymatic activities [113]. For instance, although single- or double-strand DNA repair activity has not been established for ALKBH8, the presence of RNA binding and methyltransferase domains in ALKBH8 (Figure 9) suggested that this homolog plays a role in maintenance of methylation patterns. Investigation of such activities led to the identification of ALKBH8 tRNA methyltransferase activity, necessary in the biogenesis of wobble uridine modifications utilized in translational decoding [126, 127].
Figure 9.
AlkB family protein domain alignment. Conserved amino acid sequences and domain function are indicated. The total number of amino acids is indicated to the right of each homolog. [110,113,125]
4.3.2. Substrate recognition/repair mechanism
Initially, it was predicted that AlkB family proteins directly repaired alkylation adducts by hydroxylating methyl groups and removing the resultant hydroxymethyl groups via an oxidative reaction that directly restores the undamaged base [94, 109, 112, 124, 128, 129]. However, specific investigation of the AlkB family dealkylation mechanism [130] determined that the direct repair reaction mediated by AlkB family proteins involves several intermediate steps that had not yet been identified. Regardless, dealkylation catalyzed by AlkB and its human homologs occurs via transformation of α-ketoglutarate into succinate, formaldehyde release, and restoration of the undamaged base [94, 100, 111, 130, 131] (Figure 10).
Figure 10.
ALKBH protein repair reactions. (A) ALKBH methyl adduct repair reaction. (B) ALKBH ethyl adduct repair reaction. (C) ALKBH etheno adduct repair reaction. Repair of ethyl and etheno adducts requires the same co-factors, but displaces acetaldehyde or water and glyoxal as byproducts of the repair reaction, respectively, instead of formaldehyde [100,102,103]
First, Fe(II) and three water molecules must be coordinated within the conserved catalytic core, stimulating α-ketoglutarate (KG) binding in the catalytic pocket. Binding of α-KG into the catalytic pocket chelates Fe(II) by displacing two water molecules to create the Fe(II)/α-KG active-site complex. Ligation of dioxygen to the Fe(II) molecule displaces the remaining water molecule, generating a ferric-superoxido species that undergoes self-redox and nucleophilic attack on the α-keto group. This nucleophilic attack is necessary to decarboxylate α-KG, releasing succinate and generating a ferryl-oxo intermediate. Reorientation of this intermediate facilitates removal of a hydrogen atom from the methyl adduct. Finally, radical rebound hydroxylation of the methylene group results in decomposition of the hydroxymethyl nucleobase, yielding formaldehyde and the repaired nucleobase. Though two co-factors were noted initially, α-ketoglutarate and Fe(II), ascorbate also plays a role, helping to convert the Fe(III) to Fe(II), thereby regenerating the original oxidative state of iron in the Alkbh proteins that permits enzymatic cycling [94, 111, 112, 122, 124, 130].
The major methylated bases repaired by ALKBH proteins are 1-methyladenine (1-meA) and 3-methylcytosine (3-meC), however homologs have also been reported to repair ethylated, and some etheno and exocyclic bases [102-105, 107, 131, 132]. Similar mechanisms are proposed for repair of ethano and exocyclic etheno (ε) adducts, though the final steps of these reactions result in release of acetylaldehyde and glycol, respectively [130] (Figure 10). However, additional biochemical studies are needed to confirm these mechanisms in similar detail to removal of methyl adducts from DNA.
4.3.3. Gene expression/protein regulation
Human AlkB DNA repair homologs, ALKBH2 and ALKBH3 are single genes on chromosomes 12q24 and 11p11, respectively. Expression of human AlkB homologs has been reported in a variety of normal tissue samples, including ALKBH homologs 4-8, despite the lack of DNA repair activity in the literature [133]. Expression of ALKBH family proteins varies depending on cell types. Protein expression levels in the various tissue types vary depending on the homolog evaluated. Little is known of ALKBH protein regulation mechanisms and is an area in need of further study.
4.3.4. Protein localization and cell type dependence
Differences amongst AlkB homolog proteins in their biological roles are partially ascribed to their sub-cellular localizations. ALKBH2 and ALKBH3 homolog proteins are expressed at the highest levels in the testis and ovary, however detectable expression of all AlkB homolog proteins is exhibited in the spleen, pancreas, lung, kidney, prostate and brain [133]. Although ALKBH1 activity is confined to mitochondria [115], immunofluorescence imaging indicates that the protein is cytoplasmic and nuclear [133]. Similarly, AlkB homolog proteins ALKBH3, 4, 6, and 7 are also present in the nucleus and cytoplasm [133], though ALKBH3 is the only homolog reported to possess repair activity [1, 104, 111]. Localization of ALKBH3 in both the nucleus and cytoplasm are consistent with identified interactions with helicase enzymes to facilitate DNA repair [134] and roles in mRNA repair [131]. ALKBH2 is present only in the nucleus and exhibits diffuse as well as localized, punctate staining, supporting pre-established co-localization with PCNA at replication foci during S phase [111, 131, 133], suggesting a role in replication- and transcription-related repair, as well as genome maintenance. On the contrary, AlkB homolog proteins ALKBH5 and 8 are present only in the cytoplasm [133], which supports known ALKBH8 tRNA methyltransferase activity [126, 127].
4.3.5. Post-translational modification
Unlike MGMT, ALKBH proteins are not suicide enzymes and a single protein can catalyze multiple direct repair reactions, requiring only ascorbate to regenerate the Fe(II) active site center [135]. Therefore, immediate degradation of ALKBH proteins following repair is not required, as it is for MGMT. Other possible post-translational modifications in ALKBH2 and ALKBH3 include candidate sites for phosphorylation and acetylation. Mass-spectrometric analysis of a curated database of cell lines revealed that both ALKBH2 and ALKBH3 proteins undergo post-translational modification of specific residues present in various cancer types [92].
Post-translational modifications curated for ALKBH2 include acetylation of residue Lys34 and Lys104 in various colorectal cancer cell types (HCT116, HT29, XY3-92-T and XY3-68-T), as well as phosphorylation of residue Thr252 in esophageal cancer cell line XY2-E111N [92]. Though the exact effects of these modifications are unknown, it is important to state that Lys34 is within the variable region of the N-terminus that is thought to provide protein specificity. Similarly, Lys104 is between two residues that make contact with the complimentary DNA strand during double-strand DNA repair and Thr230 is a residue in the most C-terminal α-helix of the active site [92]. Examination of potential ALKBH2 modification sites using predictive software shows possible post-translational modification sites for methylation (Arg128, 135), sumoylation (Lys75, 205), and ubiquitination (Lys104), along with other possible phosphorylation sites (Ser36, 56, 130, 182, 202, 206, 208; Thr37; Tyr91, 115) [90-93]. All of those possible post-translational modifications merit further consideration.
Post-translational modifications were also present in ALKBH3, corresponding to various disease states. Phosphorylation of Thr126 and Tyr127 residues in the β-hairpin of the NRL, as well as residue Try229 in the ALKBH3 active site, was present in acute myelogenous, chronic myelogenous and/or T-cell leukemia [92]. Additionally, phosphorylation of Tyr127 was exhibited in lung and non-small cell lung cancer cell lines. Phosphorylation of residue Tyr143, which precedes the first residue of the second β-hairpin in the NRL, was also noted in the gastric carcinoma cell line MKN-45, as well as phosphorylation of residues T212 and T214, within the ALKBH3 active site, was found in liver cancer tissue samples [92]. Examination of potential ALKBH3 modification sites using predictive software shows possible post-translational modification sites for acetylation (Lys43, 116, 219, 220), and sumoylation (Lys57, 236), along with other possible phosphorylation sites (Ser32, 50, 187, 192, 208, 265; Thr29, 41; Tyr78, 127, 229) [90-93]. All of those possible post-translational modifications merit further consideration.
5. Biological significance of direct repair in mammalian cells
Normal cells depend on direct repair to eliminate damage that is possibly cytotoxic or mutagenic. Our knowledge of the biological significance of direct repair proteins in mammalian cells is based on the evaluation of effects on cell cytotoxicity, replication, transcription and subsequent mutagenic consequences observed in the absence of each protein of interest. Recent investigations performed in model system organisms, most prominently in mice, to assess the impact of the absence of Mgmt or Alkbh family proteins will be highlighted in this section. These studies also provide insight into the function and importance of direct repair proteins in humans.
5.1. Knock-out animal models
It is important to remember that a number of DNA repair systems are implicated in the elimination of DNA lesions formed by exposure to alkylating agents. Therefore, dysfunction of repair systems can lead to pathologies that include cancer development. However, without use of a model organism to assay the effects, the consequences to the organism as a whole cannot be assessed. Knock-out animal models are a valuable tool for understanding the overall physiological effects of genes on an organism, and provide insight into disease research and therapeutic development.
Murine Mgmt models have been studied by multiple groups to evaluate sensitivity to alkylating agents commonly used in chemotherapeutics [5, 6, 82, 86, 136-139]. Though Mgmt repairs DNA damage that is known to be mutagenic, Mgmt-deficient mice surprisingly lack any overt phenotype. However, these mice are significantly more sensitive to treatment with N-methyl-N-nitrosourea (MNU), 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2-chloroethyl)3-nitrosourea (ACNU), streptozotocin, temozolomide, and dacarbazine alkylating agents [5, 136, 137, 139-142]. Mgmt knock-out mice treated with various chemotherapeutic agents also show ablation of hematopoietic tissues at the stem cell level [38, 141, 143] and are prone to development of thymic lymphomas [144]and lung adenomas [82, 138, 144, 145]. Similarly, mouse embryonic stem (ES), embryonic fibroblasts (MEFs) and bone marrow cells deficient in Mgmt also exhibit a significant increase in sensitivity (~10-fold) to MNU and BCNU [83, 141, 146]. However, mice heterozygous for Mgmt do not display a significant reduction in survival following treatment with nitrosoureas or increased tumorigenesis, compared to their wild-type counterparts.
Although in vitro DNA repair activity has been established for ALKBH1, studies conducted in murine models lacking Alkbh1 suggest roles involved in transcription. Mice deficient in Alkbh1 exhibit apoptosis in adult testis, sex-ratio distortion and unilaterial eye defects, as well as impaired differentiation of specific trophoblast lineages in the developing placenta [147, 148]. Though the specific activity and function of ALKBH1 remains to be determined, ALKBH1 biological roles seem linked to spermatogenesis and embryonic development.
On the other hand, Alkbh2- and/or Alkbh3-deficient murine models do not manifest any obvious phenotype or histopathological changes [116, 119, 132]. However, over time mice lacking Alkbh2 accumulate significant levels of 1-meA, confirming a role in removing endogenous DNA alkyl adducts. In a recent study, Alkbh2,Alkbh3,Aag knock-out mice (Aag also known as Mpg, a DNA glycosylase in the BER pathway) were viable, but underwent rapid death when exposed to a chemically-induced colitis treatment [119]. Similarly, primary mouse embryonic fibroblasts (MEFs) derived from mice lacking functional Alkbh2 exhibited significantly increased cytotoxicity and mutagenesis following exposure to the SN2 alkylating agent methyl methanesulfonate (MMS) [116, 118, 119]. Survival of Alkbh3-deficient MEFs exposed to MMS was reduced by ~50% compared to wild type MEF sensitivity, though mutant frequency did not significantly increase [116].
5.2. Replication and transcription defects
Though not all lesions generated by exposure to alkylating agents cause defects in replication and transcription, DNA and RNA adducts that are specifically removed via a direct repair mechanism interfere with replication and transcription machinery. The presence of O6-meG in DNA impedes polymerization by DNA and RNA polymerases [31, 32, 149, 150]. Polymerase beta (β), involved in base excision repair (BER) of alkylation adducts, is completely blocked by O6-meG adducts [150]. Polymerase delta (δ) is able to replicate past, but insertion of the correct base opposite O6-methylguanine is very inefficient. However, these adducts can be bypassed using polymerase eta (η) [149], a member of the Y-family DNA translesion synthesis (TLS) polymerases, but TLS polymerases are notorious for being error-prone. Interestingly, when replicating past O6-meG DNA adducts, TLS polymerase, Polη is twice as efficient at inserting cytosines opposite O6-meG as replicative polymerase, Pol δ [32].
1-meA and 3-meC lesions that are repaired by Alkbh2 and Alkbh3 are at DNA base-pairing positions and hinder proper base insertion [101]. During replication, this can lead to arrest of nucleotide synthesis, resulting in replication fork collapse [151]. Similarly, 1-meA and 3-meC adducts can also cause stalling of transcription. Correspondingly, Alkbh2 co-localizes with replication foci during S-phase [111, 131, 133] and Alkbh3 has a role in removal of alkyl adducts from mRNA [1, 15, 108, 115, 131, 152]. However, a TLS polymerase that is linked to 1-meA and/or 3-meC DNA adduct bypass has not been identified.
5.3. Cell cytotoxicity
Treatment with alkylating agents introduces a variety of adducts into DNA and RNA (Figure 2, Table 1). In the absence of direct repair proteins, those lesions can lead to cell death or damage tolerance, which allows for cell survival, but can introduce mutations into the genome that could have detrimental effects [101, 116, 142, 153]. As exhibited in Mgmt- and Alkbh-deficient murine models, lack of direct repair proteins correlates with a significant increase in cell death following treatment with SN1 or SN2 alkylating agents, respectively [116, 118, 140, 141].
5.4. Mutagenesis
When a modified nucleoside can form at least two hydrogen bonds, transcription and replication templates and translation of messengers are active [13]. O6-meG, 1-meA, and 3-meC are all involved in DNA base-pairing. Modification at O6-meG and 3-meC still allow for formation of two hydrogen bonds, while 1-meA results in only a single hydrogen bond between paired bases [13]. However, the exocyclic amino group of 1-meA can rotate so that both amino group hydrogen molecules can generate the necessary base-pairing bonds, though a slight distortion of the double-strand DNA helix does occur [13]. The addition of a methyl group to O6-G, N1-A, or N3-C interferes with normal replication, and could recruit DNA translesion synthesis (TLS) polymerases to bypass the DNA adducts. The size and organization of the Y-family TLS polymerase active sites is variable and allows for accommodation of numerous adducts. However, not only are TLS polymerases inherently error-prone [154, 155], the number and type of hydrogen bonds that can be made with the modified bases has been altered. Those factors can produce insertion of an erroneous base during bypass that accompanies replication or transcription.
O6-meG mutagenicity has been established in bacterial and mammalian systems [29, 30]. O6-meG is mutagenic and primarily gives rise to G:C→A:T mutations. A mis-insertion of thymine is thought to occur due to mis-identification of O6-meG as adenine, as hydrogen bonding can occur with the N1 and exocyclic amino group of O6-meG [13].
Unfortunately, studies evaluating the mutagenicity of a site-specific 1-meA, 3-meC, 1-meG, or 3-meT adducts have not been conducted in mammalian systems, but studies in E. coli, show that 1-meA adducts are only slightly mutagenic, whereas 3-meC, 1-meG, and 3-meT adducts are much more mutagenic [101]. Work evaluating the anti-mutagenic role of Alkbh2 and Alkbh3 in a murine model showed increased mutant frequency, specifically for mouse embryonic fibroblast (MEF) cells deficient in either Alkbh2 or Alkbh3 [116]. Those Alkbh-deficient cells exhibited increased amounts of C:G→A:T C:G→T:A mutations, respectively. Additionally, when treated with MMS, Alkbh2-deficient MEFs displayed an increased frequency of C:G→T:A and T:A→A:T mutations. Similarly, Alkbh3-deficient MEFs also exhibited an increased frequency of T:A→A:T mutations, as well as an increased frequency of A:T→G:C mutations, in response to MMS treatment. Like O6-meG, misidentification of the modified DNA bases due to the presence of two sites for hydrogen bond formation could arise if 1-meG or 3-meC is recognized as thymine and an adenine is paired with the two remaining hydrogen bond acceptors. Furthermore, T:A→A:T mutations could arise if 3-meT becomes recognized as adenine and a thymine is paired via hydrogen bonds between thymine O4 and O2 and adenine N-3 exocyclic amino group nitrogen. It is likely that 1-meA is rarely mutagenic in E. coli, deficient in AlkB, because 1-meA can utilize the C6 exocyclic amine and N7 as an alternative binding site providing two sites for hydrogen bond formation with thymine N-7 and O4 molecules, using Hoogsteen base-pairing [156].
6. Medical significance of direct repair proteins in humans
Genetic and epigenetic controls that regulate MGMT, ALKBH2, and ALKBH3 gene expression and influence how these proteins directly repair DNA are critical factors that can lead to a better understanding of cancer development. In addition, comprehension of factors that cause variations in the direct DNA repair activities of cancer cells will provide important progress toward formulating cancer therapeutics that target MGMT or ALKBH proteins. Understanding the impact of direct DNA repair proteins will eventually result in treatments that can be tailored to achieve better therapeutic results or to predict treatment and/or disease outcomes.
6.1. Epigenetic and transcriptional regulation
Epigenetic modifications are stable alterations of DNA that are heritable in the short term, but do not involve mutations of the DNA itself, and are mediated by DNA methylation and histone modifications. The stable alterations that are involved in epigenetics have a major role in exerting control on gene expression. Endogenous cell signaling as well as external influences, including diet and other life style choices, can alter gene expression mediated by changes in epigenetic modifications [157, 158]. Methylation of cytosines at transcription factor recognition sites can interfere with binding and/or function and repress transcription of that gene [159, 160]. Alternatively, protein recruitment that binds methyl CpG islands can block transcription machinery or alter chromatin structure [161, 162]. Transcriptional silencing also is connected to histone deacetylation [163, 164]. Methyl CpG binding domain (MBD) family proteins direct histone deacetylases to remove acetyl groups from lysines in the amino terminal histone tails, stabilizing DNA-histone interactions, and condensing chromatin so that transcription factor binding sites are inaccessible.
Though unmethylated in normal cells, transcriptional silencing of MGMT, associated with promoter CpG island methylation has been reported in a variety of cancer cell types and MGMT-deficient cell lines [82, 138]. Additionally, in a glioma mouse model a subpopulation of glioma cells with stem cell properties were identified [165] that are capable of re-establishing tumor growth following temozolomide treatment. Although Mgmt promoter CpG methylation or protein levels were not determined in that study, when MGMT transcript levels were evaluated in glioma patients [166], those with MGMT CpG promoter methylation had increased response to temozolomide, but also maintained a subset of glioma cells with stem cell-like character and MGMT promoter methylation. Interestingly, mRNA levels of DNMT1 and DNMT3b methyltransferases are increased in a number of human glioma patients, but there does not appear to be a link to MGMT expression levels [167]. Moreover, MGMT promoter CpG methylation levels and DNA methyltransferase levels alone do not account for patient response to alkylating agent therapy. However, whether MGMT promoter methylation disables transcription factor binding or contributes to chromatin reorganization remains uncertain [71, 72, 74]. Therefore, regulation of MGMT expression is still unclear and merits intense scrutiny.
The inability to establish direct connections among MGMT expression, CpG methylation, and response to alkylating agent therapy indicates that other mechanisms contribute in regulating MGMT levels. Studies evaluating MGMT expression and microRNAs in patient samples have established a modest inverse correlation between the levels of MGMT transcript and miR-181d [168]. Moreover, expression of mi-181d in A1207 glioblastoma cells, results in abnormal sensitivity to temozolomide. However, expression of MGMT cDNA, restores the survival to levels close to that of the A1207 parental line. These results suggest that identification of other miRNAs involved in regulating MGMT expression will help elucidate the mechanisms that control the gene transcript levels.
In addition to control at the DNA and transcript levels, histone modifications can also control the epigenetic state and direct expression. Acetylated histone H3 and H4 levels also increase in cell lines expressing MGMT, compared to cell lines deficient in MGMT [169], which would facilitate nucleosomal positioning that enables transcription factor interactions. Further, binding of MBD proteins in the MGMT promoter of was greater in MGMT-silenced cells, implicating MBD proteins in recruitment of histone deactylases that remove lysine acetylation from the amino-terminal tails of histones H3 and H4, resulting in more condensed chromatin and transcription inactivation [73, 79, 170]. Therefore, epigenetic and/or enzymatic CpG island methylation at the MGMT promoter influences transcription factor access, as well as chromatin structure that are important for MGMT expression.
ALKBH2 and ALKBH3 both have CpG islands in their promoters, but epigenetic regulation and/or gene silencing has not been reported for either homolog. However, mutations that alter protein expression have been observed [171], but it is likely that methylation of CpG islands near any of the seven transcription factor binding sites in the promoter of ALKBH2 or the single transcription factor binding sites within the promoter region of ALKBH3, would repress transcription factor binding and possibly gene expression. Because data on the function of ALKBH promoters are less abundant compared to those available for the MGMT promoter, examination of the promoter function for those genes is an area that would benefit from further investigation.
6.2. Links to cancer
Dysregulation of numerous DNA repair pathways are involved in tumor development, progression, diagnosis, treatment and prognosis, including direct DNA repair proteins [82, 159, 172-179]. Over-expression of direct repair proteins is generally associated with a protective effect against cell death that would otherwise be induced by alkylating agent treatment. However, down-regulation or silencing of direct repair protein expression is associated with increased mutagenesis that precedes tumorgenesis. Therefore expression profiles could be used to predict potential resistance or enhanced sensitivity to therapeutics.
MGMT has been implicated in many types of human tumors. Numerous MGMT polymorphisms have risk associations with breast, lung, colon, and head and neck cancers [63, 82, 180-186]. Decreased MGMT expression is also found in glioma, lymphoma, retinoblastoma, breast (including triple-negative breast cancer) and prostate cancer [82, 138, 187] [188]. Moreover, lack of MGMT is associated with enhanced outcomes using alkylating agent therapies [5, 62, 67, 82, 86, 138, 139, 180, 181, 183, 189]. Though MGMT silencing occurs in a variety of tumor types, increased levels have also been observed in non-Hodgkin lymphoma, myeloma and glioma, as well as in some colon, pancreatic, breast, and lung cancers [63, 183, 184].
Mutations in ALKBH2 and 3 have been associated with an enhanced expression of these proteins in glioma cells and pediatric brain tumors [171, 190]. Similarly, over-expression of ALKBH3 has been associated with human rectal carcinoma [191] and prostate cancer, as well as, lung adenocarcinoma and non-small-cell lung cancer [134] [192]. On the contrary, down regulation of ALKBH2 has been observed in gastric cancer, promoting growth of gastric cancer cells [193]. Although down regulation of ALKBH2 in gastric cancer cells caused increased proliferation, ALKBH2 silencing in H1299 lung cancer cells had the opposite effect, increasing cisplatin sensitivity. Similarly, ALKBH3 silencing induced senescence and sensitivity to alkylating agents in human adenocarcinoma and prostate cancer cells [134, 193]. Therefore, further study of the role of ALKBH2 and 3 in both normal and tumor cells is necessary to elucidate their biological role(s).
6.3. Therapeutic targets
Understanding the mechanism of proteins involved in various DNA repair pathways is crucial for developing new chemotherapeutic targets and eventually new drugs. DNA alkylating agents and ionizing radiation (IR) are often used as chemotherapeutic treatments because of ability to control the dose administered and area of treatment, as well as the major cytotoxic effects of both agents at high doses. However, in addition to generation of cytotoxic adducts that cause apoptosis, alkylating agents and IR also form adducts that can be mutagenic and as a result can cause initiation of secondary cancers. Although DNA repair deficiencies are associated with increased cancer risk and formation, cancer cells proficient in DNA repair can reduce therapeutic efficacy. Currently, combination cancer treatment regimens are being explored that utilize chemotherapy or IR and target specific DNA repair proteins with pharmacological agents to enhance treatment efficacy and eliminate resistance to treatment regimens exhibited in some patients [189].
6.3.1. MGMT
Chemotherapeutic drugs such as temozolamide (TMZ) and bis-(2-chloroethyl)-nitrosourea (BCNU) generate some lesions repaired via the direct methyltransferase mechanism. Combination treatment with MGMT inhibitors prevents repair and resistance to methylating and chloroethylating agents [1, 38, 137] and has also been shown to reverse cisplatin drug resistance [194].
Understanding cellular regulation of MGMT expression will allow for selective down regulation and sensitization of tumors to alkylating agent chemotherapies. Studies have evaluated manipulation of MGMT expression and protein levels. Initial experiments evaluating MGMT inhibitors identified O6-benzyl guanine (BG) as an efficacious inhibitor of MGMT activity, a single, micromolar dose depleting greater than 99% of MGMT activity in human cells for 24-hours following drug removal [195]. Moreover, treatment with BG lacks any mutagenic or cytotoxic effects [195-197]. Clinical trials combining BG and BCNU treatment have been conducted in colon cancer, sarcoma, melanoma and myeloma, as well as studies evaluating combination of BG and TMZ [138]. Since synthesis of BG, additional BG-like inhibitors have been developed [196], including O6-(4-bromothenyl) guanine, which has been evaluated in patients with glioma [187]. Similarly, targeting of MGMT along with combination of platinum drugs, including cis- and carboplatinum [198], as well as topoisomerase I inhibitors has been investigated in various clinical trials [86].
Another approach to regulate MGMT that holds great, essentially untapped therapeutic potential is strategies utilizing RNA interference-mediated gene silencing to target MGMT [168, 199, 200]. For instance, if anti-sense molecules can specifically target MGMT mRNA translation, and degradation is also inhibited, depletion of MGMT is sustainable for long periods of time [62]. As seen in glioblastoma patients, expression levels of various miRNA markers correlate with prognosis [168, 199, 200]. Therefore, one potential new treatment could use miRNAs, such as miR-181d, to decrease MGMT levels, thus increasing sensitivity to alkylating agents [168]. Similarly, targeting regions of the MGMT promoter that is accessible to transcription factors could interfere with binding and down-regulate MGMT transcription. However, non-specific targeting of MGMT inhibitors in all cells increases chemotherapeutic toxicity. Therefore, mutant forms of MGMT that are resistant to BG-like inhibitors are also being evaluated to limit myelosuppression, affording hematopoietic progenitor cells protection from BG and BCNU or temozolomide treatment [201-204].
6.3.2. Alkbh homologs
Similar to MGMT, the role of ALKBH2 and ALKBH3 in repair of DNA alkylation damage at base-pairing sites is anti-carcinogenic. However, investigations indicate that over-expression of ALKBH proteins in various cancer cell lines shields those cells against methylating agent toxicity and would thereby protect against some chemotherapeutic treatments [134, 171, 192]. Additionally, because loss of ALKBH2 and/or ALKBH3 leads to disruption of replication, inhibition of ALKBH2 and/or ALKBH3 is a strong target for the development of novel chemotherapeutic agents. Some specific inhibitors of these proteins have already been identified [135, 205, 206], as well as generic α-KG/dioxygenase inhibitors including dimethyl oxalylglycine (DMOG) and α-ketoglutarate derivatives such as oxoglutarate. Studies have addressed the application of DNA aptamers as inhibitors of ALKBH proteins [207]. However, to date no studies have been conducted in mammalian models that evaluate the combination of ALKBH inhibitors with chemotherapeutic alkylating agents.
7. Summary
Direct repair proteins represent a unique class of enzymes that remove DNA damage without a dependence on DNA synthesis. In the future, better comprehension of how these proteins function and are produced in cells will lead to understanding their roles in formation of mutations that cause cancer. Eventually, that knowledge will foster the development of drugs to target these proteins and/or to regulate their expression to improve patient outcomes.
\n',keywords:null,chapterPDFUrl:"https://cdn.intechopen.com/pdfs/44566.pdf",chapterXML:"https://mts.intechopen.com/source/xml/44566.xml",downloadPdfUrl:"/chapter/pdf-download/44566",previewPdfUrl:"/chapter/pdf-preview/44566",totalDownloads:3337,totalViews:684,totalCrossrefCites:0,totalDimensionsCites:2,totalAltmetricsMentions:1,impactScore:0,impactScorePercentile:5,impactScoreQuartile:1,hasAltmetrics:1,dateSubmitted:"April 17th 2012",dateReviewed:"October 16th 2012",datePrePublished:null,datePublished:"May 22nd 2013",dateFinished:"May 7th 2013",readingETA:"0",abstract:null,reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/44566",risUrl:"/chapter/ris/44566",book:{id:"3302",slug:"new-research-directions-in-dna-repair"},signatures:"Stephanie L. Nay and Timothy R. O‘Connor",authors:[{id:"155613",title:"Dr.",name:"Timothy",middleName:null,surname:"O\\'Connor",fullName:"Timothy O\\'Connor",slug:"timothy-o'connor",email:"toconnor@coh.org",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"City Of Hope National Medical Center",institutionURL:null,country:{name:"United States of America"}}},{id:"157190",title:"BSc.",name:"Stephanie",middleName:null,surname:"Nay",fullName:"Stephanie Nay",slug:"stephanie-nay",email:"snay@coh.org",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Direct repair substrates—DNA and RNA alkylation damage",level:"1"},{id:"sec_3",title:"3. Sources of alkylation damage",level:"1"},{id:"sec_3_2",title:"3.1. Types of alkylating agents",level:"2"},{id:"sec_4_2",title:"3.2. DNA and RNA alkylation damage",level:"2"},{id:"sec_6",title:"4. Direct repair proteins",level:"1"},{id:"sec_6_2",title:"4.1. Mechanisms of alkylation repair",level:"2"},{id:"sec_7_2",title:"4.2. Methyl Guanine Methyl Transferase (MGMT) proteins",level:"2"},{id:"sec_7_3",title:"4.2.1. Protein structure/active site organization",level:"3"},{id:"sec_8_3",title:"4.2.2. Substrate recognition/repair mechanism",level:"3"},{id:"sec_9_3",title:"4.2.3. Gene expression/protein regulation",level:"3"},{id:"sec_10_3",title:"4.2.4. Protein localization and cell type dependence",level:"3"},{id:"sec_11_3",title:"4.2.5. Post-translational modification",level:"3"},{id:"sec_13_2",title:"4.3. Alkbh Fe(II)/α-ketoglutarate-dependent dioxygenases",level:"2"},{id:"sec_13_3",title:"4.3.1. Protein structure/active site organization",level:"3"},{id:"sec_14_3",title:"4.3.2. Substrate recognition/repair mechanism",level:"3"},{id:"sec_15_3",title:"4.3.3. Gene expression/protein regulation",level:"3"},{id:"sec_16_3",title:"4.3.4. Protein localization and cell type dependence",level:"3"},{id:"sec_17_3",title:"4.3.5. Post-translational modification",level:"3"},{id:"sec_20",title:"5. Biological significance of direct repair in mammalian cells ",level:"1"},{id:"sec_20_2",title:"5.1. Knock-out animal models",level:"2"},{id:"sec_21_2",title:"5.2. Replication and transcription defects",level:"2"},{id:"sec_22_2",title:"5.3. Cell cytotoxicity",level:"2"},{id:"sec_23_2",title:"5.4. Mutagenesis",level:"2"},{id:"sec_25",title:"6. Medical significance of direct repair proteins in humans",level:"1"},{id:"sec_25_2",title:"6.1. Epigenetic and transcriptional regulation",level:"2"},{id:"sec_26_2",title:"6.2. Links to cancer",level:"2"},{id:"sec_27_2",title:"6.3. Therapeutic targets",level:"2"},{id:"sec_27_3",title:"6.3.1. MGMT",level:"3"},{id:"sec_28_3",title:"6.3.2. Alkbh homologs",level:"3"},{id:"sec_31",title:"7. 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Chemoprotective gene transfer II: multilineage in vivo protection of haemopoiesis. Gene Ther. 1998;5(6):842-7.'},{id:"B202",body:'Hickson I, Fairbairn LJ, Chinnasamy N, Lashford LS, Thatcher N, Margison GP, et al. Chemoprotective gene transfer I: transduction of human haemopoietic progenitors. Gene Ther. 1998;5(6):835-41.'},{id:"B203",body:'Koc ON, Reese JS, Davis BM, Liu L, Majczenko KJ, Gerson SL. DeltaMGMT-transduced bone marrow infusion increases tolerance to O6-benzylguanine. Hum Gene Ther. 1999;10(6):1021-30.'},{id:"B204",body:'Reese JS, Koc ON, Lee KM, Liu L, Allay JA, Phillips WP, Jr., et al. Retroviral transduction of a mutant methylguanine DNA methyltransferase gene into. Proc Natl Acad Sci U S A. 1996;93(24):14088-93.'},{id:"B205",body:'Karkhanina AA, Mecinovic J, Musheev MU, Krylova SM, Petrov AP, Hewitson KS, et al. Direct analysis of enzyme-catalyzed DNA demethylation. Anal Chem. 2009;81(14):5871-5.'},{id:"B206",body:'Woon EC, Demetriades M, Bagg EAL, Aik WS, Krylova SM, Ma JHY, et al. Dynamic combinatorial mass spectrometry leads to inhibitors of a. J Med Chem. 2012;55(5):2173-84.'},{id:"B207",body:'Krylova SM, Koshkin V, Bagg E, Schofield CJ, Krylov SN. Mechanistic studies on the application of DNA aptamers as inhibitors of. J Med Chem. 2012;55(7):3546-52.'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Stephanie L. Nay",address:null,affiliation:'
Irell and Manella Graduate School of Biological Sciences, USA
Department of Cancer Biology, Beckman Research Institute, Duarte, CA, USA
'},{corresp:null,contributorFullName:"Timothy R. O‘Connor",address:null,affiliation:'
Department of Cancer Biology, Beckman Research Institute, Duarte, CA, USA
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1. Introduction
Discourse, a construct with the personal thought which reflects personal behavior and attitude, is known as the ideology. The socially conditioned and socially constructed ideas are considered as ideology. Ideology connects peoples’ perspectives from the different fields, such as, in the context of Nepal, political leaders may develop one kind of thought; on the other hand, educationists may develop educational thoughts differently. The perspectives educationist and the political leaders create the interactive situation in which they could discuss their thoughts and generate new thoughts. Fairclough [1] identifies transforming goals with the perspectives of textual and contextual variations; discursive practices may have significant ideological effects, which they can help to produce and reproduce unequal power relations among the different bodies of the community and how they represent the things and position of people. Van Dijk [2] states that ideologies are the ideas and belief system of a particular group of people defined from the multidisciplinary ways involving social, cognitive, and discursive aspects. He further argues that ideologies are acquired, expressed, changed, and reproduced in the society, mainly in different forms of discourses such as texts and talks.
According to Fairclough and Wodak [3], “CDA as discourse analysis which aims to systematically explore the often opaque relationship of causality and determination between discursive practices, events, texts and wider social and cultural structures, relations and processes; to investigate how such practices, events, and texts arise out of and are ideologically shaped by the relation of power and struggles overpower, and to explore how the opacity of this relationship between discourse and society is itself a factor securing power and hegemony.”
Regarding the above definition, ideology refers to social forms of processes within which and employing which, symbolic forms circulate in the social world. Ideology is the study of “how meaning is constructed and conveyed by symbolic forms of various kinds” [4]. Another perspective suggested by Luke [5] in a different context claims that language gets power when influential people use it. There is a close relationship between language and power in several ways; societal and cultural frameworks shape the ideological interaction which is based on the language associated with power relationship.
In Refs. [6, 7], it is stated that ideology is perceived as a multidisciplinary approach which is the fundamental framework for organizing multiple ideological concepts and consumptions. Similarly, Van Dijk [6] focuses that ideologies usually control the thoughts of a social group which then represent the essential social characteristics of a group based on their identities, goals, norms, values, positions, and responses to other negative stances. However, Van Dijk [2] emphasizes the ideological consumption in both cognitive and social levels and proclaims that social cognition is a system with shared sociocultural knowledge by members of a specific group, society, or culture. On the other hand, cognitive functions are considered as the basic form of ideological properties that organize, monitor, and control attitudes of a social group and are accompanied by the experiences of a person [6].
Ideology represents the discursiveness by which it allows interaction in the correspondence of the social thoughts and responsibilities. As suggested by Van Dijk [8], ideologies are particular ways of representing and constructing a society that reproduces distributed power relations.
Ideology is not only associated with representing social reality. It reflects the social identity and construction of thought. Therefore, ideology imparts the presence of thoughts coordinated by social and cultural influences.
2. Basic traits
Ideologies are foundational social beliefs of people rather than general in nature because fundamental ideologies are not developed overnight, such as a person cannot be a feminist or socialist in few days but it takes time for framing the foundation of personal, social, and contextual ideologies including many experiences and discourses. Hodge [9] states that interaction is a basic requirement for discourse which is sturdy and transformative as it introduces different perspectives, ideas, structures, and ontologies. Ideologies always interact in different forms. For example, Van Dijk [6] contends that ideologies are endorsed in the forms of action and interaction, and ideological reproduction is often rooted in organizational and institutional contexts. For example, racist ideologies are expressed in racist talk, and feminists reproduce their ideologies in feminist talk. Therefore, many forms of interactional discourse play a vital role in communicating ideologies in the society. Ideologies are the principles that essentially function as the cognitive representations in the form of discourse, societal position, and interests of social groups which connect macrolevel analyses of social structure with microlevel studies of individual interaction. However, Martínez-Roldán and Malavé [10] contend that ideologies are not only the shared beliefs of a group, social interactions between individuals within a sociocultural context, and negotiation of meanings but also indicate multifaceted social phenomena including various social classes, groups, and social institutions within the broader societal contexts. It involves diverse social groups within different contradictory ideological groups.
The context of ideology reflects the personal framework [4]. Moreover, Fairclough and Wodak [3] state that context has the determining role in producing the ideology. In the context of communication, simply we get a contradiction of the ideologies. For example, media discourse may have a different ideology to educational and political discourses. The tenets of ideological interaction theory represent the guidelines to present the contextual variations or perspectives, as suggested in Refs. [1, 7, 8, 9]. The basic tenets of critical discourse analysis introduced by Fairclough and Wodak [3] are the baseline for determining the traits of ideological interaction perspectives.
Social dimension has the decisive capacity to determine the ideological interaction
Ideological creation in the perspectives
Culture, sex, social status, and economic status are discursive
Contextual discourses and communication abilities and skills are core thought of the ideology
3. The framework of ideological interaction theory
The framework of ideological interaction theory is based on sociocultural perspectives. Scott and Palincsar [11] argued that cultural and social entities suggest ideological consequences. Similarly, in Refs. [12, 13], the significance of the cultural contribution to implant ideology is stated, as an ideology has an impact on the social activities, attitudes, and shared understanding in interdisciplinary fields. The social role has tribute consideration in determining the ideology. For example, the authors in Refs. [2, 3, 4, 5, 7, 9, 14] focus on the contextual, cognitive, and attribute relevance and variable to construct the thought, and the thought is considered to the social issue; in the way, social issues may have multidimensional and diverse perspectives might be revealed as addressing the ideology.
Ideologies are expressed by text and talks as mentioned by Padilla and Vana [15]. It helps to construct new and confirm already present ideologies persuasively. Similarly, the communication skills and strategies lead to construct ideologies and prevail interactions among groups in different circumstances.
On the other hand, there are unseen priori theoretical grounds to exclude any textual structures from expressing underlying ideological principles [3]. Undeniably, the mental model of functional categories involved in events or communicative contexts.
Some of the tenets of critical discourse analysis can already be found in the critical theory of the Frankfurt School before the Second World War [3].
The framework discusses on the interaction process where society and language interact to the individual and cultural awareness and that have the effects of critical discourse approaches from the baseline, and with this reference, an individual perceives a perspective of a discourse of any educational or political situation. Moreover, the different levels of the discourses interact to the ideologies, and by the product, there is another form of ideology. For example, the Modern Era introduces diverse thoughts and beliefs guided by ideology and conscience, and more or less, they are interacting with the other ideologies around us. In this process of interactions, we generate other modified thesis and new perspectives to see things around us. As proof, we can take an example from the situation of the COVID-19 pandemic. In Nepal, the suicidal death rate is 25 times higher than the death of those infected during this lockdown period. People have different perspectives. Some people argue that the financial crisis has caused this, whereas other people believe that it happened because of fear and psychological threat; but after the interaction between these two diverse ideologies, people have come to modify their arguments by considering each other’s factual arguments. Now, the state of their ideologies shares some common ground and they have generated ideology based on their thought and social status which is the outcome of the ideological interaction theory.
4. Issues and contexts
The theory of ideology is articulated within a conceptual triangle [6] that informs the discourse analytic approach and connects society, discourse, and social cognition in the framework of a critical discourse analysis. Ideologies are the straightforward outlines for establishing the social cognition communal by associates of social groups, organizations, or institutions. In this respect, ideologies are both cognitive and social [6, 7, 16]. Moreover, ideologies function and interface the interface between the cognitive representations and processes underlying discourse and action, on the one hand, and the societal position and interests of social groups, on the other hand. Similarly, according to Padilla and Vana [15], this conception of ideology also allows us to establish the crucial link between macrolevel analyses of groups, social formations, and social structure and microlevel studies of situated, individual interaction, and discourse. Ideologies, then, are the overall, abstract mental systems that organize such socially shared attitudes. In Refs. [9, 17], the authors report in the different contexts that the feminist attitudes just mentioned, for instance, which may be internally structured and mutually related by general principles or propositions that together define a feminist ideology. Similar examples may be given for racist, anti-racist, corporate, or ecological attitudes and their underlying ideological systems.
As stated by Vygotsky [18], sociocultural issues play a vital role in the formation of discourses, and scaffolding creates several opportunities to interact with the different behaviors in society. On the other hand, ideological configurations are landed by the approaches of critical discourse analysis [12]. However, Lazar [19] states that the approach, specifically, feminism shows the level of interaction among the communities and personal perspectives. The way we perceive the concept is crucial for shaping the grounded reality. Scott and Palincsar [11], on the other hand, discuss the sociocultural impacts on discourse management.
Here I am presenting some examples of interaction connected to feminism and sociocultural realities.
Feminist theory focuses on analyzing the nature of gender inequality, women’s social roles, interests, choices, and female politics in the different fields such as in philosophy, sociology, psychology, literature, and education [20]. It examines the exploitation, domination, sexual objectification, oppression, patriarchy, and stereotyping. The theory of feminism mainly talks about the role of women in society, such as being economically dependent, having low status and power, doing unpaid work, loss of choice, and division of labor in different cultural and sexist beliefs.
Feminism theory first appeared in about 1794 from the United States, and it evolved dealing with contemporary issues and women’s experiences [21]. There are different concepts associated with the theory of feminism. According to Code [20], there are different waves in feminism; the first wave was initiated in about 1928 which argued for the political equality and the rights of property and representation of women in the society, by some of the leading figures of feminism such as Wollstonecraft and Suffragettes.
Similarly, the second wave of feminism started after the Second World War period, representing the female emancipation and woman liberation movement of the 1950s and 1960s which mainly argued for the social and economic equality of the women in the society, especially equal pay and equal rights. It reacted against the uneven distribution of power, women’s racial, religious, class-related, and different forms of oppressions. The third wave appeared by pointing out some of the drawbacks of second-wave feminism in the late 1980s and 1990s by representing the experiences of woman globally—women of different races, classes, ethnicities, and sexualities are positioned differently within the countries. The most recent wave of feminism deals with the female empowerment of women in the use of digital technology or social media to encourage female achievement.
While analyzing the transcribed text of Muniba’s video speech, this research connects feminism which covers the gender-based realities in the society to its transformation to social activities. The critical discourses value the diverse perspectives in the society based on the concept of power, domination, and social equality focusing on groups, organizations, and institutions. Personal and cultural knowledge may have the crucial role in ideological interaction [19]. He further says that group knowledge is shared by the social group of society. Reality lies in the knowledge and attitude representing the ideologies of social groups such as socialists, ecologists, feminists, as well as anti-feminists and can be analyzed in terms of local and global interaction among groups and social organizations.
Throughout analyzing this discourse, the term feminism refers not only to female-oriented characteristics but also to gender practices with constructing values. Similarly, the concept reflects gender identity, responsibility, belief, and the traditions of the social practices concerning the ground reality of women in the society. Lazar [22] reports that feminism is one of the critical perspectives of social transformation and emancipation with the values of gender-based realities that existed in society. Further, he argues that the society has the dividend role from the perspectives of gender and that focuses to the categories of cultural influences, social identity, sexuality, discrimination, power practices, social position, ideology; particularly, asymmetrical power relations and gender ideology are the major discussions of feminism. Urbain [23] discusses the feminist movement from society to pedagogy; the main concern lies in the core aspects of the feminist movement, care, diversity, and collaboration.
The video that I selected has a good connection to the disability and material feminist theory; for example, Mays [24] discusses domestic violence, social oppression as a moment of gender and disability dimensions. The psychosociological context has several emerging issues of feminism; however, the real situation that the disabled female has been coping with in the sociocultural setting demands exploring gender and disability dimensions.
In the speech, she has described about how people in the society including her relatives behave her after being physically disable in a car accident and how she dealt with socially diversified ideologies to live a successful and motivating life despite her physical inability. As we know, feminism focuses on gender identity with the aspects of society as well as cultural configurations. Van Dijk [25] discusses several approaches regarding the socio-cognitive perspectives where feminism remains as the core component. He further supports that social inequalities have been introducing several hidden realities such as attainment of knowledge of diverse groups or issues, practicing the social and cultural values associated with the ideological awareness, and social equity in the society.
For the exploration of the issue material, feminist interpretations and disability theory associated with cultural dimension have the prominent focus to experience the gendered and disabling capacities in the sociocultural environment. The women are biased due to the male dominance in the process of decision-making and power relations. Some questions such as: Are there no provision of women’s independence? Is there no value to women’s decisions? Why society marks a huge gap between male and female for education, job, and lifestyle? triggered my mind while observing the nature of society. I wanted to introduce and explore the different aspects of society where females live. I found the speech motivating to describe the hidden potentialities of the women who want to introduce their existence and prove their perfection in their physical imperfection. This empowers all the women around the world to recognize their inner capacities, to face problems created by the social system, patriarchy, and power relations. It is observed in the speech that despite many challenges, the females can raise their voices to unlock the more significant opportunities and possibilities in society.
The speech is full of positive and negative feelings of pain, sorrows, fear, and determination. She has unhappy feelings about being disabled in the accident due to her belief in misfortune and fate. She explained the situation as follows:
The next day, the doctor came to me and said, “Your spine injury is so bad you won’t be able to walk again.” I took a deep breath. And I said it’s all right. The next day doctor came and said, “Because of your spine injury and the fixation that you have in your back, you won’t be able to give birth to a child again.” That day, I was devastated [26].
The women with disabilities are depicted as weak, helpless, vulnerable, dependent, and incapable bodies [27]; the doctor had reported her as unable to give birth and walk normally. The word devastated illuminates her deep and sad feeling of disappointment, hopelessness, and unhappiness.
In another context, Mazari [26] opines, “I’m tired of looking at these white walls in the hospital and wearing these white scrubs. Bring me some colours, bring me some small canvas. I want to paint.” The word tired of is related to her unhappiness as she was convicted for full 2 years in the complete bed rest and “I want to paint” shows that she wanted to overcome all those negativities and forget the pain by painting.
According to her, “So the very first painting I made was on my deathbed where I painted for the very first time, it was not just an art piece or just my passion. It was my therapy.” To signify that she expresses her internal pain into that painting as she has a passion for portraying herself in the beautiful arts. In this way, she develops a feeling and emotion of fear, sadness, and negativity to determination. The feeling of determination has overcome her reality when she says,
After 2 years and two and a half months when I was able to sit in a wheelchair, that was the day when I had the rebirth. So, I have to accept myself the way I am, the sooner the better.
This shows her strong sense of accepting herself as the way she is. Finally, she tried to motivate the audience to summarize all her life experiences and the struggle she faced by sharing some happy and successful moments in her expressions. She says, “I became the national goodwill ambassador for UN Women, Pakistan. And now I speak for the rights of women and children. We talk about inclusion, diversity, gender equality which is a must.”
Mays [24] argued that women with disabilities experience social domination and domestic violence continuously as a result of gender and disability dimensions. It is praiseworthy and inspirational to all people that despite all these miseries, illness, injuries, hatred, ignorance, disability, and lack of acceptance, she overcame her depressing feelings and faced the world as an average and happy person. By accepting all the challenges and personal limitations, how she managed her life to return in a new transformative form shows her a motivational and robust figure in the world. Muniba wanted to convey that every bad experience teaches us the best lessons in life, when she says, in the first line of her speech “They see my disability, I see my ability,’ they call me to disable, I call me differently able.” The words delivered by her reflect her strong sense of self-determination and confidence. She takes the suffering of spinal cord injury as a challenge and becomes more determined to express her feelings through her art and paintings. She further suggests to all the audience and says,
Live your life fully, accept yourself the way you are, be kind to yourself, and only you can be kind to others, love yourself and spread the love if you accept the way are, the world will recognize you, it’s all starts from within.
Through her speech, she has proven to the world as a capable and inspiring lady despite her disability. She is spreading motivation to all the people around the world to have a positive sense of self-acceptance and kindness to the self before spreading compassion to the world.
The cultural aspects of feminism believe in gender equality, and radical feminism believes that women are dominated by the practice of patriarchy in the family [28]. Mazari expresses that she was 18 years old when she got married. She said, “if that makes you happy, I will say ‘YES.’ And of course, it was never a happy marriage.” This has characterized how social structure and patriarchy are the root cause of gender inequality leading women to domination. This exemplifies how she accepted the forceful marriage proposal led by her father though she was not happy in the marital relationship. Similarly, she said, “You know what was my biggest fear? ”Divorce. I couldn’t stand this word. I was trying to cling on to this person who didn’t want me anymore I got the news that he is getting married, I sent him a text and said, ‘I am so happy for you and I wish you all the best.’ This demonstrates how men have the freedom and choice in their life of making a decision of remarriage, ignoring the disabled wife in the pain and miseries. This depicts the real picture of male domination over females.
From the linguistic perspective, the language, and utterance, linguistic aspects such as grammar, vocabulary, cohesion, and structure used in the speech of Muniva Mazari were analyzed, which were found as per the principle of consistency and comprehensiveness.
The motivational discourse of Muniba Mazari illustrates that if people want to explore their hidden talents regardless of their physical disabilities, social inequalities, domination, and weaknesses, they can transform into a recognized person contributing to the society, nation, and the world. Human life is full of challenges, hardship, and turmoil but a strong sense of determination makes people break all those barriers. Disability is not a limitation to those who take it as an opportunity to explore their inner capacities. Ignorance and inequalities make victim suffer a lot but finally makes a person more strong-minded and goal-oriented as Muniba Mazari who has been working for many social campaigns, spreading awareness on gender discrimination, women, and child rights.
Muniba’s speech is the reflection of the societal structure including gender discrimination, violence, domination, and ignorance of the disability. Some of the important realities she exposed in her speech are how women are living in the society, how their decision-making affects their life, how they react to the unexpected incidents in life, how the disable people need familial and social acceptance, how females are standing in their career, and how they feel being discriminated and ignored by the society and dear ones. Her speech reflects personal experiences representing life before and after disability. The speaker focuses on motivating people, particularly women, to recognize self-power of potentialities, to spread love, care, and kindness to the self and the rest of the world by fighting against social and gender discriminations.
In another context, critical discourse analysis deals with multiple perspectives guided by ideology, power, social activities, action, and ethics. Kalina and Powell [12] argue that the approaches of the constructivist thoughts have been recognized as the best practices with multiple meanings around the globe. Constructivism has several branches; among them, the sociocultural theory of Vygotsky might have a good connection with the discourse. Turuk [29] opines that human understanding is the fundamental concept of sociocultural theory as it connects with different levels of connections such as zone of proximal development (ZPD). Vygotsky [30] claims that a child is a dependent phenomenon, and the sociocultural environment ensures the engagement of the task, activities, and instructions to actualize the shape of learning.
Sociocultural theory reflects the sociocultural influences as well as relationship traits in learning behaviors. The meaning and learning situation is situated in social values and cultural traditions. In the same way, the theory introduces the collaborative and interactive situation. In the context of personality development, Jaramillo [31] explains that learners’ success depend upon the collaboration. Similarly, sociocultural theory creates spaces for learner’s active participation and motives with regard to the unit of the developmental framework, and the letter written by Lincoln demands the social and cultural setting regarding child psychology.
On the other hand, Vygotsky [18] claims that sociocultural theory differs from cognitive approaches, in that, the cognitive approaches assign several internal processes to gain the knowledge gaining channels with reference to socially as well as culturally mediated prospects. Kalina and Powell [12] support the model of sociocultural learning modality and focus on the collaborative act, coordination skills, negotiation strategies, and creative as well as critical behaviors for the successful adaptation of learning behaviors.
Therefore, the theory focuses to the gap between the inter-psychological/social and intra-psychological/individual aspects and believes that collaborative and psycho-centric instruction helps learners to understand and see how interactions take place and enable learners to achieve the goals within a social instructional network; with the assumption, I used this theory, though others’ theories are also equally possible to connect with the text that I have selected for the analysis.
Lincoln’s primary concern is that the teacher should be able to instruct his child on the thought that educational scholarship should accompany the first instruction of character. He took education as a journey to gain values of faith, love, and courage and asked the teacher to teach his son moral and ethical values as his son could realize the world and his responsibility. These values square measure ones that transcend content learning and facilities to make the premise of one’s identity. Lincoln desires the teacher to reflect the truth that education is character based. Similarly, he instructs his son’s principal to show his son a way to be a decent, honorable, and ethical person.
Furthermore, he wants his son to avoid jealousy. He further needs his son to understand each learning from a book and enjoying the refinement and wonder of nature. He needs his son to follow his integrity and to find out to track what he thinks is true, though it is unpopular. He should moreover learn that it’s additionally honorable to fail than to comprehend accomplishment through dishonesty. Lincoln needs his son to feel comfortable to express unhappiness and nevertheless to find out to laugh once he is unhappy. Lincoln asks the principal to be kind to his son, however, do not spoil him and show his son patience. In the end, Lincoln needs his son to find out to be on his own.
In my observation, I found that Lincoln connected learner to social beings and focused the learners’ reality in two basics, such as the right learning and wrong learning. As discussed in the letter, right being leads to the social well-being and to developing social thoughts and responsibilities, with reference to enhancing positive as well as cooperative and collaborative learning goals. For instance, education is defined as values, love, faith, courage, ethics, responsible thoughts, and personal as well as social identity and character. On the other hand, wrong being made the learners to tackle the situation and that might lead to the development of leadership skills.
The cultural reflection is another part of the letter, as the writer wanted to see his son as one of the examples of the cultural entity in which he could sketch his introduction and independence in society. Hence, he imagined a positive, motivated, encouraged, determined, dedicated, diligent, problem-oriented, and socially responsible son and that could be shaped by the teachers only. In his letter, he discussed honesty, positivity, and sublime faith on own self as the three fundamental values. According to him, honesty develops a positive mind, leads to happiness, and develops a realistic view. Similarly, positivity encourages us to be cheerful and prepares to face the challenges. He further claims that sublime faith itself enables learners to have sublime faith in humanity.
Similarly, knowledge reveals the several facets of the common goals such as content development, social responsibility, cultural well-being, political insights, and emotional radicals. According to the letter, Lincoln highly emphasized the values that he expected from the teachers.
The letter is the representation of the social guideline as to the triangularity (learners, teachers, and parents) with the diverse nature of social and cultural rhetorics. The letter is an example of the expectation of parents from the teachers, as they want their children to be accustomed to the diverse realities of the multidimensional world. It mainly focuses on the determination of individual principles concerning the sociocultural perspectives in the multidisciplinary rationale of transformational ethics. The letter covers several areas like psychology, sociology, linguistics, pedagogy, and other various disciplines and approaches. The writer only focused on the teacher’s responsibility as concerning the issue, and it would be better if he could incorporate parents’ role at home as students are at school for some limited time. Consequently, he needs to shortly talk on the home environment that he had been providing to his son. Therefore, every parent should play a vital role and be responsible for shaping the future of their children as they need to deal with different realities and the people around them.
Eventually, the letter illustrates and prioritizes that education is not only about teaching the books and using materials, but it should be practical and connected with the realities of society and the world too. This educational discourse is a mirror of the society that reflects how education should be incorporated in practical and real life.
This letter proves that education is a multidisciplinary approach that not only includes teaching and learning inside the four walls, but it is a process of habit formation, teaching students the real values and norms connecting with the social behaviors and real world outside. In Ref. [32], it is argued that Lincoln believed losing something is connected with learning because it is another way out or looking for alternatives to get another path to proceed with the task.
Regarding the sociocultural perspectives, learners’ motivation, teachers’ responsibility, parents’ expectations, and societal needs are associated with forming cultural identities. Therefore, the letter reflects the social changes and adaptation. However, the learners are expected to be found to the determining curriculum, and they might develop their insights through the hidden curriculum or natural orders of learning praxis.
In the above contexts, generally, we find the interaction of the ideology with the individual and society.
5. Conclusion
Frankfort school introduces the relationship between theory and society, social inquiry, and the interconnection of the social life with the economy, psychology, ethics, traditions, values, and focus on social and historical contexts [33]. In the same vein, Kim [16] argues that the primary consideration is the transformation of knowledge. The knowledge has radical consequences, and subjective and intersubjective practices are implied.
Social framework and cultural consciousness are in the central concern for the study of social inquiries and activities. Critical consciousness, cultural inheritance, and sociopolitical adaptation and aspirations are critical considerations in educational practices.
In Refs. [15, 34], the authors believe on multidisciplinary and transdisciplinary educational policy in the field of critical pedagogy. It has a connection to ideology, power, economics, politics, and knowledge diversity. Discursive practice, social phenomena, and cultural diversity are the key issues discussed in the articles. Weiss and Wodak [7] suggest critical pedagogy, value system, power relations, critical consciousness, and critical literacy rather than functional literacy to promote a subjective form of knowledge, culture, and understanding. Following Refs. [4, 5, 8, 35, 36], the role of education should be for developing a critical consciousness; furthermore, the educational approaches are highly context-sensitive and democratic approaches take an ethical stance on social issues to transform society into an approach or attitude rather than a step method. The socio-transformative practices’ impact is seen accompanied by sociopolitical, socio-historic contexts which contribute to the production and interpretation of the text and are crucial aspects of the analysis. However, social responsibility connected with ideological concern is crucial for critical pedagogy, and power relations determine that. The societal framework is the connecting paradigm where different social theories and approaches surround the mind, ideology, and perspectives.
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The concern criteria are ideology and the other social components like people, status, economy, media, and politics with their connectivity to the global situational trends. 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Introduction",level:"1"},{id:"sec_2",title:"2. Basic traits",level:"1"},{id:"sec_3",title:"3. The framework of ideological interaction theory",level:"1"},{id:"sec_4",title:"4. Issues and contexts",level:"1"},{id:"sec_5",title:"5. Conclusion",level:"1"}],chapterReferences:[{id:"B1",body:'Fairclough NL. Critical and descriptive goals in discourse analysis. Journal of Pragmatics. 1985;9(6):739-763'},{id:"B2",body:'Van Dijk TA. Ideology and discourse analysis. Journal of Political Ideologies. 2006;11(2):115-140'},{id:"B3",body:'Fairclough N, Mulderrig J, Wodak R. Critical discourse analysis. In: Dijk TAV, editor. Discourse Studies: A Multidisciplinary Introduction. Los Angeles: Sage; 1997. pp. 357-378'},{id:"B4",body:'Thompson JB. Ideology and Modern Culture: Critical Social Theory in the Era of Mass Communication. Hoboken NJ, United states: John Wiley & Sons; 2013'},{id:"B5",body:'Luke A. Beyond science and ideology critique: Developments in critical discourse analysis. 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Critical Theory in Education: Philosophical, Research, Sociobehavioral, and Organizational Assumptions. Mexico: Eastern New Mexico University; 2000'},{id:"B14",body:'Van Dijk TA. 18 critical discourse analysis. In: The Handbook of Discourse Analysis. 2001. pp. 349-371'},{id:"B15",body:'Padilla LV, Vana R. Ideologies in the foreign language curriculum: Insights from textbooks and instructor interviews. Language Awareness. 2019;28(1):15-30'},{id:"B16",body:'Kim AI. (De)legitimation of monolingual ideologies in a US teachers’ online forum. International Journal of Bilingual Education and Bilingualism; 2020:1-12'},{id:"B17",body:'Ayers DF. Neoliberal ideology in community college mission statements: A critical discourse analysis. The Review of Higher Education. 2005;28(4):527-549'},{id:"B18",body:'Vygotsky L. Interaction between learning and development. Readings on the Development of Children. 1978;23(3):34-41'},{id:"B19",body:'Lazar MM. Feminist critical discourse analysis. In: Handbook of Critical Discourse Studies. 2017. pp. 372-387'},{id:"B20",body:'Code L. Encyclopedia of Feminist Theories. New York, NY: Taylor & Francis e-Library; 2000'},{id:"B21",body:'Wilkinson S, Kitzinger C. Heterosexuality: A Feminism & Psychology Reader. Sage; 1993'},{id:"B22",body:'Lazar MM. Politicizing gender in discourse: Feminist critical discourse analysis as political perspective and praxis. In: Feminist Critical Discourse Analysis. Springer; 2005. pp. 1-28'},{id:"B23",body:'Urbain M. A Feminist Critical Discourse Analysis of the National Board for Professional Teaching Standards. 2018'},{id:"B24",body:'Mays JM. Feminist disability theory: Domestic violence against women with a disability. Disability & Society. 2006;21(2):147-158'},{id:"B25",body:'Van Dijk TA. 18 critical discourse analysis. In: The Handbook of Discourse Analysis. 2001. p. 352'},{id:"B26",body:'Mazari M. Muniba Mazari the inspiring “Iron Lady of Pakistan” [YouTube Video]; 2017'},{id:"B27",body:'Garland-Thomson R. Integrating disability, transforming feminist theory. NWSA Journal. 2002:1-32'},{id:"B28",body:'Campbell R, Wasco SM. Feminist Approaches to Social Science: Epistemological and Methodological Tenets. 2000'},{id:"B29",body:'Turuk MC. The relevance and implications of Vygotsky’s sociocultural theory in the second language classroom. Arecls. 2008;5(1):244-262'},{id:"B30",body:'Vygotsky L. Socio-cultural theory. In: Mind in Society. 1978'},{id:"B31",body:'Jaramillo JA. Vygotsky’s sociocultural theory and contributions to the development of constructivist curricula. Education. 1996;117(1):133-141'},{id:"B32",body:'Lessons from Abraham Lincoln’s Letter to his Son’s Headmaster [Internet]. 2018. Available from: https://medium.com/@dinhnguyen0504/abraham-lincolns-letter-to-his-son-s-headmaster-2f287fd33bfa'},{id:"B33",body:'Martin J, Jay ME. Splinters in Your Eye: Essays on the Frankfurt School. Verso; 2020'},{id:"B34",body:'Pais A, Costa M. An ideology critique of global citizenship education. Critical Studies in Education. 2020;61(1):1-16'},{id:"B35",body:'Bachmann V, Moisio S. Towards a constructive critical geopolitics–Inspirations from the Frankfurt School of critical theory. Environment and Planning C: Politics and Space. 2020;38(2):251-268'},{id:"B36",body:'Portschy J. Times of power, knowledge and critique in the work of Foucault. Time and Society. 2020;29(2):392-419'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"Yadu Prasad Gyawali",address:"gyawaliyadu.gyawali@gmail.com",affiliation:'
Mid-Western University, Surkhet, Nepal
'}],corrections:null},book:{id:"6944",type:"book",title:"Heritage",subtitle:null,fullTitle:"Heritage",slug:"heritage",publishedDate:"September 9th 2020",bookSignature:"Daniela Turcanu-Carutiu",coverURL:"https://cdn.intechopen.com/books/images_new/6944.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",isbn:"978-1-83881-925-5",printIsbn:"978-1-83881-924-8",pdfIsbn:"978-1-83881-926-2",isAvailableForWebshopOrdering:!0,editors:[{id:"176482",title:"Prof.",name:"Daniela",middleName:null,surname:"Turcanu-Carutiu",slug:"daniela-turcanu-carutiu",fullName:"Daniela Turcanu-Carutiu"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}}},profile:{item:{id:"192432",title:"Dr.",name:"Carlos",middleName:null,surname:"Rosales",email:"carosal@biomedicas.unam.mx",fullName:"Carlos Rosales",slug:"carlos-rosales",position:null,biography:null,institutionString:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",totalCites:0,totalChapterViews:"0",outsideEditionCount:0,totalAuthoredChapters:"3",totalEditedBooks:"0",personalWebsiteURL:null,twitterURL:null,linkedinURL:null,institution:{name:"National Autonomous University of Mexico",institutionURL:null,country:{name:"Mexico"}}},booksEdited:[],chaptersAuthored:[{id:"53789",title:"Cellular and Molecular Mechanisms of Insect Immunity",slug:"cellular-and-molecular-mechanisms-of-insect-immunity",abstract:"Multicellular organisms constantly encounter potentially harmful microorganisms. Although insects lack an adaptive immune system, they do have powerful means of fighting infections. Cellular responses involve phagocytosis of bacteria and encapsulation of parasites. In addition, insects can mount a humoral response against pathogens. This is characterized by the secretion of antimicrobial peptides into the hemolymph. Recognition of foreign pathogens involves specific receptors for sensing infection. These include peptidoglycan recognition proteins (PGRPs) and β‐glucan recognition proteins (βGRPs). Engagement of these receptors starts signaling pathways that activate the genes that encode antimicrobial peptides. These pathways include the Toll, the Imd, and the JAK‐STAT. This chapter describes the innate immunity of insects including both the cellular and humoral responses to bacteria, fungi, and parasites. In addition, recent advances in insect antivirus immune responses are discussed.",signatures:"Carlos Rosales",authors:[{id:"192432",title:"Dr.",name:"Carlos",surname:"Rosales",fullName:"Carlos Rosales",slug:"carlos-rosales",email:"carosal@biomedicas.unam.mx"}],book:{id:"6048",title:"Insect Physiology and Ecology",slug:"insect-physiology-and-ecology",productType:{id:"1",title:"Edited Volume"}}},{id:"54543",title:"Neutrophil Role in Periodontal Disease",slug:"neutrophil-role-in-periodontal-disease",abstract:"Oral tissues are constantly exposed to damage from the mechanical effort of eating and from the invasion of foreign microorganisms such as bacteria, fungi, and virus. In healthy oral tissues, there is a balance between symbiotic bacteria and cells from the innate immune system, mainly neutrophils. When this balance is broken, inflammation appears and more immune cells are recruited to the gingiva. Neutrophils form a barrier against dysbiotic bacteria. However, when neutrophils are insufficient, bacteria thrive causing periodontitis, a chronic inflammatory disease that destroys the tooth‐supporting tissues or periodontium. Damage of periodontal tissues leads to tooth loss, and in severe cases, it can also affect systemic health by increasing a person's risk for atherosclerosis, rheumatoid arthritis, diabetes, and even cancer. The mechanisms neutrophil employ to keep a balance with bacteria in order to maintain healthy oral tissues is the focus of this chapter. We discuss how neutrophil antimicrobial functions keep bacteria at check and how some dysbiotic bacteria block neutrophils to promote an inflammatory state. Also, novel therapeutic approaches for periodontitis are discussed.",signatures:"Carlos Rosales and Eileen Uribe‐Querol",authors:[{id:"192432",title:"Dr.",name:"Carlos",surname:"Rosales",fullName:"Carlos Rosales",slug:"carlos-rosales",email:"carosal@biomedicas.unam.mx"},{id:"198687",title:"Dr.",name:"Eileen",surname:"Uribe-Querol",fullName:"Eileen Uribe-Querol",slug:"eileen-uribe-querol",email:"euquerol@comunidad.unam.mx"}],book:{id:"5834",title:"Role of Neutrophils in Disease Pathogenesis",slug:"role-of-neutrophils-in-disease-pathogenesis",productType:{id:"1",title:"Edited Volume"}}},{id:"63248",title:"Neutrophil Activation by Antibody Receptors",slug:"neutrophil-activation-by-antibody-receptors",abstract:"Neutrophils, the most abundant leukocytes in blood, are relevant cells of both the innate and the adaptive immune system. Immunoglobulin (Ig) G antibody molecules are crucial activators of neutrophils. IgGs identify many types of pathogens via their two Fab portions and are in turn detected through their Fc portion by specific Fcγ receptors (FcγRs) on the membrane of neutrophils. Thus, antibodies bring the specificity of the adaptive immune response to the potent antimicrobial and inflammatory functions of neutrophils. Two types of FcγRs with several polymorphic variants exist on the human neutrophil. These receptors are considered to be redundant in inducing cell responses. Yet, new evidence presented in recent years on how the particular IgG subclass and the glycosylation pattern of the antibody modulate the IgG–FcγR interaction has suggested that a particular effector function may in fact be activated in response to a specific type of FcγR. In this chapter, we describe the main types of FcγRs on neutrophils and our current view on how particular FcγRs activate various signaling pathways to promote unique effector cell functions, including phagocytosis, activation of integrins, nuclear factor activation, and formation of neutrophil extracellular traps (NETs).",signatures:"Carlos Rosales and Eileen Uribe-Querol",authors:[{id:"192432",title:"Dr.",name:"Carlos",surname:"Rosales",fullName:"Carlos Rosales",slug:"carlos-rosales",email:"carosal@biomedicas.unam.mx"},{id:"198687",title:"Dr.",name:"Eileen",surname:"Uribe-Querol",fullName:"Eileen Uribe-Querol",slug:"eileen-uribe-querol",email:"euquerol@comunidad.unam.mx"}],book:{id:"7129",title:"Neutrophils",slug:"neutrophils",productType:{id:"1",title:"Edited Volume"}}}],collaborators:[{id:"176263",title:"Dr.",name:"Miroslav",surname:"Kostic",slug:"miroslav-kostic",fullName:"Miroslav Kostic",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"176326",title:"Dr.",name:"Sladjan",surname:"Stankovic",slug:"sladjan-stankovic",fullName:"Sladjan Stankovic",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"193144",title:"Dr.",name:"Monica",surname:"Rosenblueth",slug:"monica-rosenblueth",fullName:"Monica Rosenblueth",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"National Autonomous University of Mexico",institutionURL:null,country:{name:"Mexico"}}},{id:"195706",title:"Dr.",name:"Tania",surname:"Rosas-Pérez",slug:"tania-rosas-perez",fullName:"Tania Rosas-Pérez",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"195709",title:"Mr.",name:"Arturo",surname:"Vera-Ponce De León",slug:"arturo-vera-ponce-de-leon",fullName:"Arturo Vera-Ponce De León",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"195710",title:"Dr.",name:"Shamayim T.",surname:"Ramírez-Puebla",slug:"shamayim-t.-ramirez-puebla",fullName:"Shamayim T. Ramírez-Puebla",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"195711",title:"Dr.",name:"Reiner",surname:"Rincón-Rosales",slug:"reiner-rincon-rosales",fullName:"Reiner Rincón-Rosales",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"195712",title:"Mr.",name:"Julio",surname:"Martínez-Romero",slug:"julio-martinez-romero",fullName:"Julio Martínez-Romero",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"195713",title:"Dr.",name:"Michael F.",surname:"Dunn",slug:"michael-f.-dunn",fullName:"Michael F. Dunn",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"195714",title:"Dr.",name:"Esperanza",surname:"Martínez-Romero",slug:"esperanza-martinez-romero",fullName:"Esperanza Martínez-Romero",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null}]},generic:{page:{slug:"orders-and-delivery",title:"Order and Delivery Info",intro:'
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Our books are available hardcover, printed in full colour and produced to the highest standards on PEFC™ and FSC certified paper, complying with principles of responsible forestry worldwide. The paper size is 180 x 260 mm (7 x 10.2 inches).
\n\n
Print On Demand (POD)
\n\n
\n\t
IntechOpen Books are printed specifically for your order
\n\t
Ordered, printed, and delivered in 7-15 business days
\n\t
Available for purchase at any time no minimum or maximum threshold on book order quantity
\n
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IntechOpen works with award winning print-houses and we hold to the fact that all of our printed products are of the highest quality.
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Prices and Discounts
\n\n
IntechOpen books retail price range is:
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100 - 159 GBP ex. VAT (available in USD and EUR)
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Discounts available:
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All IntechOpen contributors can buy the print copies of books for an Author Exclusive price with discounts from 30% to 50% on retail price. Log in to your Author Panel to purchase a book at the discounted price.
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Libraries are offered a 20% discount.
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Bulk discounts are granted for orders of 10 copies and more.
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There is no minimum or maximum threshold on the quantity of book orders.
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Orders have to be paid in advance and before printing. We accept payment in GBP, EUR and USD.
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IntechOpen will help you complete your payment safely and securely, keeping your personal, professional and financial information safe.
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In accordance with the best security practice, we do not accept card orders via email.
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The combined printing and delivery time for orders vary from 7-15 business days, depending on the printed quantity and destination. This period does not include any customs clearance difficulties that may arise and that are beyond our control. Once your order has been printed and shipped, you will receive a confirmation email that includes your DHL tracking number. You can then track your order at www.dhl.com.
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If you do not receive your order within 30 days from the date your order is shipped, please contact us to inquire about the shipping status at orders@intechopen.com.
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Customs: free shipping does not include any duties, taxes or clearing charges levied by the destination country. These charges are the responsibility of the customer and will vary from country to country.
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IntechOpen partners do not provide shipping service from Europe to the countries listed below. Please refrain from mailing items addressed to the countries listed below, until further notice.
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When ordering our books from the countries listed below, please provide an alternative mailing address. For any further assistance, please contact us at orders@intechopen.com.
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Representatives
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Print copies of our publications are most often purchased by universities, libraries, institutions and academia personnel, hence increasing the visibility and outreach of our authors' published work among science communities and institutions.
\n\n
Our books are available at our direct Print Sales Department and through selected representatives throughout the world.
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Books International
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Representative for: Brunei, Cambodia, Indonesia, Indonesia, Laos, Malaysia, Myanmar, Philippines, Singapore, Thailand, Vietnam (ASEAN)
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LSR Libros Servicios y Representaciones S.A. de C.V
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Unexpected delays in construction projects are caused by internal and external environments embedding several risk factors which may occur concurrently. The cost overrun and schedule overrun not only influence the construction industry’s completion of a project but can also have profound effect on the economy of a country. Even though the failure of the construction projects to get completed within the budgeted cost and time has received attention by researchers, lot more need to be researched as to what can be done to have tight leash on construction projects so that they can be brought on track. In order to meet the stiff deadlines involving complexity of construction projects, the scheduling of projects should be flexible enough to accommodate changes without negatively affecting the overall project cost and duration. This chapter deals with Perspectives on Risk Assessment and Management Paradigms as applicable to any project in general and construction industry in particular.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"K. Srinivas",authors:[{id:"255339",title:"Prof.",name:"K.",middleName:null,surname:"Srinivas",slug:"k.-srinivas",fullName:"K. Srinivas"}]},{id:"63201",doi:"10.5772/intechopen.80640",title:"Risk Management Practices Adopted by European Financial Firms with a Mediterranean Connection",slug:"risk-management-practices-adopted-by-european-financial-firms-with-a-mediterranean-connection",totalDownloads:935,totalCrossrefCites:1,totalDimensionsCites:4,abstract:"Following the economic and financial crises, any activity involving internal controls, especially risk management, has been given more attention. With this study, we aim to contribute further to the existing literature on risk management by looking at practices adopted by financial services firms licenced in Europe with a Mediterranean connection. We used parts of a questionnaire adopted by two of the authors in another study on risk management practices adopted by Maltese financial services firms and sent it to prospective candidates who work closely within risk management, to collect our data. This resulted in 1635 participants. This data was used to (1) bring to light the mechanisms and strategies used in risk management by these organisations to maximise their opportunities, manage their risks, and maintain stability in their financials. Also, (2) we check if this is perceived as contributing to ‘principled performance’. Finally, (3) we examine the extent to which risk management capabilities offer a competitive advantage to these firms. Our findings evidence that the objective by EMP and the EU, that is to ensure that members operate ‘on the same level playing field’ within risk management, in financial services of firms with a Euro-Mediterranean connection, has been achieved.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"Simon Grima and Frank Bezzina",authors:[{id:"257099",title:"Dr.",name:"Simon",middleName:null,surname:"Grima",slug:"simon-grima",fullName:"Simon Grima"},{id:"257101",title:"Prof.",name:"Frank",middleName:null,surname:"Bezzina",slug:"frank-bezzina",fullName:"Frank Bezzina"}]},{id:"76927",doi:"10.5772/intechopen.97809",title:"Integrating Resilience in Time-based Dependency Analysis: A Large-Scale Case Study for Urban Critical Infrastructures",slug:"integrating-resilience-in-time-based-dependency-analysis-a-large-scale-case-study-for-urban-critical",totalDownloads:210,totalCrossrefCites:2,totalDimensionsCites:2,abstract:"As critical systems shall withstand different types of perturbations affecting their functionalities and their service level, resilience is a very important requirement. Especially in an urban critical infrastructures where the occurrence of natural events may influence the state of other dependent infrastructures from various different sectors, the overall resilience of such infrastructures against large scale failures is even more important. When a perturbation occurs in a system, the quality (level) of the service provided by the affected system will be reduced and a recovery phase will be triggered to restore the system to its normal operation level. According to the implemented recovery controls, the restoration phase may follow a different growth model. This paper extends a previous time-based dependency risk analysis methodology by integrating and assessing the effect of recovery controls. The main goal is to dynamically assess the evolution of recovery over time, in order to identify how the expected recovery plans will eventually affect the overall risk of the critical paths. The proposed recovery-aware time-based dependency analysis methodology was integrated into the CIPCast Decision Support System that enables risk forecast due to natural events to identify vulnerable and disrupted assets (e.g., electric substations, telecommunication components) and measure the expected risk paths. Thus, CIPCast can be valuable to Critical Infrastructure Operators and other Emergency Managers involved in a crisis assessment to evaluate the effect of natural and anthropic threats affecting critical assets and plan proper countermeasures to reduce the overall risk of degradation of services. The proposed methodology is evaluated in a real scenario, which utilizes several infrastructures and Points of Interest of the city of Rome.",book:{id:"10256",slug:"issues-on-risk-analysis-for-critical-infrastructure-protection",title:"Issues on Risk Analysis for Critical Infrastructure Protection",fullTitle:"Issues on Risk Analysis for Critical Infrastructure Protection"},signatures:"Vittorio Rosato, Antonio Di Pietro, Panayiotis Kotzanikolaou, George Stergiopoulos and Giulio Smedile",authors:[{id:"27002",title:"Dr.",name:"Vittorio",middleName:null,surname:"Rosato",slug:"vittorio-rosato",fullName:"Vittorio Rosato"},{id:"284589",title:"Dr.",name:"Antonio",middleName:null,surname:"Di Pietro",slug:"antonio-di-pietro",fullName:"Antonio Di Pietro"},{id:"326117",title:"Dr.",name:"Yiorgos",middleName:null,surname:"Stergiopoulos",slug:"yiorgos-stergiopoulos",fullName:"Yiorgos Stergiopoulos"},{id:"326118",title:"Dr.",name:"Giulio",middleName:null,surname:"Smedile",slug:"giulio-smedile",fullName:"Giulio Smedile"},{id:"326119",title:"Associate Prof.",name:"Kotzanikolaou",middleName:null,surname:"Panagiotis",slug:"kotzanikolaou-panagiotis",fullName:"Kotzanikolaou Panagiotis"}]},{id:"74240",doi:"10.5772/intechopen.94917",title:"Analyzing the Cyber Risk in Critical Infrastructures",slug:"analyzing-the-cyber-risk-in-critical-infrastructures",totalDownloads:341,totalCrossrefCites:2,totalDimensionsCites:2,abstract:"Information and communication technology (ICT) plays an important role in critical infrastructures (CIs). Some ICT-based services are in itself critical for the functioning of society while other ICT elements are essential for the functioning of critical processes within CIs. Moreover, many critical processes within CIs are monitored and controlled by industrial control systems (ICS) also referred to as operational technology (OT). In line with the CI-concept, the concept of critical information infrastructure (CII) is introduced comprising both ICT and OT. It is shown that CIIs extend beyond the classical set of CIs. The risk to society due to inadvertent and deliberate CI/CII disruptions has increased due to the interrelation, complexity, and dependencies of CIs and CIIs. The cyber risk due to threats to and vulnerabilities of ICT and OT is outlined. Methods to analyze the cyber risk to CI and CII are discussed at both the organization, national, and the service chain levels. Cyber threats, threat actors, and the organizational, personnel, and technological cyber security challenges are outlined. An outlook is given to near future cyber security risk challenges, and therefore upcoming risk, stemming from (industrial) internet of things and other new cyber-embedded technologies.",book:{id:"10256",slug:"issues-on-risk-analysis-for-critical-infrastructure-protection",title:"Issues on Risk Analysis for Critical Infrastructure Protection",fullTitle:"Issues on Risk Analysis for Critical Infrastructure Protection"},signatures:"Marieke Klaver and Eric Luiijf",authors:[{id:"325818",title:"Dr.",name:"Marieke",middleName:null,surname:"Klaver",slug:"marieke-klaver",fullName:"Marieke Klaver"},{id:"325822",title:"MSc.",name:"Eric",middleName:null,surname:"Luiijf",slug:"eric-luiijf",fullName:"Eric Luiijf"}]},{id:"64655",doi:"10.5772/intechopen.82273",title:"Lifecycle Risk Modelling of Complex Projects",slug:"lifecycle-risk-modelling-of-complex-projects",totalDownloads:919,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"Large, complex and challenging engineering projects require extensive understanding and management of risk. How these risks are identified at the infancy of a project and subsequently mitigated throughout the project lifecycle is critical to successful delivery. Many projects begin with a comprehensive attempt to identify risks but lack the tools to manage and measure risk mitigation as the project progresses through the lifecycle causing the project to spiral out of control. This chapter outlines details of a risk model that uses a system within a system approach of identifying and segmenting risks. The model can then be analysed quantitatively and generate a visual lifecycle risk profile that allows the project team to monitor risks continuously in the project lifecycle. Furthermore, the use of a baseline or ideal project is proposed that is used as a measure of likely success against new projects.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"Matthew Cook and John P.T. Mo",authors:[{id:"6394",title:"Prof.",name:"John P.T.",middleName:null,surname:"Mo",slug:"john-p.t.-mo",fullName:"John P.T. Mo"},{id:"254858",title:"Ph.D. Student",name:"Matthew",middleName:null,surname:"Cook",slug:"matthew-cook",fullName:"Matthew Cook"}]}],mostDownloadedChaptersLast30Days:[{id:"64817",title:"Pharmaceutical Projects: Walking along the Risk Management Line",slug:"pharmaceutical-projects-walking-along-the-risk-management-line",totalDownloads:1261,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"We manage risk so commonly (and unconsciously) in our everyday life that we tend to undervalue it. Risk management was officially introduced in the pharmaceutical world by the ICH guideline Q9 in 2005. Since then, it has been intensively used and, not infrequently, misused. Practice shows that risk assessment tools are often seen as an end in themselves, while such important things as brainstorming on the matter and getting to know the problem are underestimated. A pharmaceutical project provides a very good example of this: risk management is critical, but as there are many unknown factors, it has to be performed in a way that what really counts is understanding the problems we face. A pharmaceutical project has at least two actors, a pharmaceutical firm and an engineering company, possessing different backgrounds, and this often leads to different approaches. This may explain why risk management is not used as much as it should in pharmaceutical projects. Thus, this chapter considers a pharmaceutical project from the point of view of risk management.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"Jordi Botet",authors:[{id:"143483",title:"Dr.",name:"Jordi",middleName:null,surname:"Botet",slug:"jordi-botet",fullName:"Jordi Botet"}]},{id:"62904",title:"Bank Risk Management: A Regulatory Perspective",slug:"bank-risk-management-a-regulatory-perspective",totalDownloads:1700,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"The globalization of financial markets, information technology development, and increasing competition have largely affected bank business and its risk management. Together with these forces, regulatory factors play a significant role. This chapter approaches bank risk management under the regulators’ perspective with an emphasis on the risk-based capital regulation. Specifically, how bank risk is regulated under the risk-based capital regulation and whether the regulation shapes bank risk are discussed in detail. In such a way, the chapter provides better understanding of the risk-based capital regulation and bank risk-taking behaviors.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"Nguyen Thi Thieu Quang and Christopher Gan",authors:[{id:"256020",title:"Prof.",name:"Christopher",middleName:null,surname:"Gan",slug:"christopher-gan",fullName:"Christopher Gan"},{id:"256021",title:"Ms.",name:"Nguyen Thi Thieu",middleName:null,surname:"Quang",slug:"nguyen-thi-thieu-quang",fullName:"Nguyen Thi Thieu Quang"}]},{id:"63163",title:"Decision-making in Risk Management",slug:"decision-making-in-risk-management",totalDownloads:1389,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"The definition of risk introduced in the ISO 31000 standard of 2009 (2018) is uncertain goal achievement; thus, both negative and positive outcomes can be considered. It also implies that risk is not limited to life and health, but may cover all goals of a company. Risk management thus becomes a question of achieving and optimizing multiple goals. Since safety is but one of several considerations, safety may lose out to other more easily measured objectives of a company, such as economics and compliance with regulatory requirements. Risk analyses have a long history of quantification, a tradition that for various reasons has waned and should be revived if safety goals are to be treated together with other goals of a company. The extended scope affects not only company owners and employees but also neighbors, the local community, and the society at large. The stochastic nature of risk and the considerable time lap between decisions and the multiattributed consequences implies that managing risk is exposed to cognitive biases of many sorts. Risk management should be based on a quantitative approach to risk analysis as a protection against the many cognitive biases likely to be present, and managers should be trained to recognize the most common cognitive biases and decision pitfalls.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"Jan Folkmann Wright",authors:[{id:"254395",title:"Mr.",name:"Jan Folkmann",middleName:null,surname:"Wright",slug:"jan-folkmann-wright",fullName:"Jan Folkmann Wright"}]},{id:"65609",title:"Risk Analysis Related to Cost and Schedule for a Bridge Construction Project",slug:"risk-analysis-related-to-cost-and-schedule-for-a-bridge-construction-project",totalDownloads:1491,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"The construction sector is subject to more risk than many other sectors. Managing risk is the hottest topic of discussion for engineers within the construction sector. It is difficult to imagine managing of projects without risk management in construction. Risk management is concerned with risk management planning, identification, analysis, responses, monitoring and controlling project risk. Risk analysis is an evaluative process that establishes the magnitude of risks on projects. This work is planned to identify and analyze risks in the construction of a bridge project. The data are collected through a survey approach by administering a questionnaire. Professionals involved in the construction of bridges identify the project risks. A case study is utilized to determine the impact of cost and schedule risks. The analysis is carried out using the Monte Carlo simulation. The findings of the Monte Carlo simulation are compared with the actual times and costs of the case-study project. The results show the actual times and costs fell within the expected distribution of the simulation. The results indicate that risk analysis is helpful in managing costs and schedule risks. Additionally, this work documents guidelines for risk analysis.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"Rafiq M. Choudhry",authors:[{id:"255346",title:"Prof.",name:"Rafiq M.",middleName:null,surname:"Choudhry",slug:"rafiq-m.-choudhry",fullName:"Rafiq M. Choudhry"}]},{id:"64630",title:"Process of Risk Management",slug:"process-of-risk-management",totalDownloads:1820,totalCrossrefCites:4,totalDimensionsCites:6,abstract:"Cost saving and timely performance are of utmost importance to all stakeholders who are involved in a construction project that is owner, contractor, consultant and subcontractor. The prime causes of risks in construction projects involve delay and failure to complete the work at specified cost and within the agreed time frame. Unexpected delays in construction projects are caused by internal and external environments embedding several risk factors which may occur concurrently. The cost overrun and schedule overrun not only influence the construction industry’s completion of a project but can also have profound effect on the economy of a country. Even though the failure of the construction projects to get completed within the budgeted cost and time has received attention by researchers, lot more need to be researched as to what can be done to have tight leash on construction projects so that they can be brought on track. In order to meet the stiff deadlines involving complexity of construction projects, the scheduling of projects should be flexible enough to accommodate changes without negatively affecting the overall project cost and duration. This chapter deals with Perspectives on Risk Assessment and Management Paradigms as applicable to any project in general and construction industry in particular.",book:{id:"7573",slug:"perspectives-on-risk-assessment-and-management-paradigms",title:"Perspectives on Risk, Assessment and Management Paradigms",fullTitle:"Perspectives on Risk, Assessment and Management Paradigms"},signatures:"K. Srinivas",authors:[{id:"255339",title:"Prof.",name:"K.",middleName:null,surname:"Srinivas",slug:"k.-srinivas",fullName:"K. 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He completed a one-year Post-Doctoral Fellowship awarded by the DFAIT (Foreign Affairs and International Trade Canada) at the Institute of Biomedical Engineering of the University of New Brunswick (Canada) in 2010. Currently, he is Professor in the Faculty of Electrical Engineering (UFU). He has authored and co-authored more than 200 peer-reviewed publications in Biomedical Engineering. He has been a researcher of The National Council for Scientific and Technological Development (CNPq-Brazil) since 2009. He has served as an ad-hoc consultant for CNPq, CAPES (Coordination for the Improvement of Higher Education Personnel), FINEP (Brazilian Innovation Agency), and other funding bodies on several occasions. He was the Secretary of the Brazilian Society of Biomedical Engineering (SBEB) from 2015 to 2016, President of SBEB (2017-2018) and Vice-President of SBEB (2019-2020). 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He is an author and co-author of scientific publications covering analysis and processing of biomedical images and development of database systems.",institutionString:"University of Silesia",institution:null}]},{type:"book",id:"6843",title:"Biomechanics",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/6843.jpg",slug:"biomechanics",publishedDate:"January 30th 2019",editedByType:"Edited by",bookSignature:"Hadi Mohammadi",hash:"85132976010be1d7f3dbd88662b785e5",volumeInSeries:4,fullTitle:"Biomechanics",editors:[{id:"212432",title:"Prof.",name:"Hadi",middleName:null,surname:"Mohammadi",slug:"hadi-mohammadi",fullName:"Hadi Mohammadi",profilePictureURL:"https://mts.intechopen.com/storage/users/212432/images/system/212432.jpeg",biography:"Dr. Hadi Mohammadi is a biomedical engineer with hands-on experience in the design and development of many engineering structures and medical devices through various projects that he has been involved in over the past twenty years. Dr. Mohammadi received his BSc. and MSc. degrees in Mechanical Engineering from Sharif University of Technology, Tehran, Iran, and his PhD. degree in Biomedical Engineering (biomaterials) from the University of Western Ontario. He was a postdoctoral trainee for almost four years at University of Calgary and Harvard Medical School. He is an industry innovator having created the technology to produce lifelike synthetic platforms that can be used for the simulation of almost all cardiovascular reconstructive surgeries. He’s been heavily involved in the design and development of cardiovascular devices and technology for the past 10 years. He is currently an Assistant Professor with the University of British Colombia, Canada.",institutionString:"University of British Columbia",institution:{name:"University of British Columbia",institutionURL:null,country:{name:"Canada"}}}]}]},openForSubmissionBooks:{paginationCount:3,paginationItems:[{id:"11580",title:"Recent Advances in Canine Medicine",coverURL:"https://cdn.intechopen.com/books/images_new/11580.jpg",hash:"1806716f60b9be14fc05682c4a912b41",secondStepPassed:!0,currentStepOfPublishingProcess:3,submissionDeadline:"March 23rd 2022",isOpenForSubmission:!0,editors:[{id:"258334",title:"Dr.",name:"Carlos Eduardo",surname:"Fonseca-Alves",slug:"carlos-eduardo-fonseca-alves",fullName:"Carlos Eduardo Fonseca-Alves"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{id:"11579",title:"Animal Welfare - New Insights",coverURL:"https://cdn.intechopen.com/books/images_new/11579.jpg",hash:"12e4f41264cbe99028655e5463fa941a",secondStepPassed:!1,currentStepOfPublishingProcess:2,submissionDeadline:"June 1st 2022",isOpenForSubmission:!0,editors:[{id:"51520",title:"Dr.",name:"Shao-Wen",surname:"Hung",slug:"shao-wen-hung",fullName:"Shao-Wen Hung"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{id:"11578",title:"Antibiotics and Probiotics in Animal Food - Impact and Regulation",coverURL:"https://cdn.intechopen.com/books/images_new/11578.jpg",hash:"3731c009f474c6ed4293f348ca7b27ac",secondStepPassed:!1,currentStepOfPublishingProcess:2,submissionDeadline:"June 3rd 2022",isOpenForSubmission:!0,editors:[{id:"225390",title:"Dr.",name:"Asghar Ali",surname:"Kamboh",slug:"asghar-ali-kamboh",fullName:"Asghar Ali Kamboh"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null}]},onlineFirstChapters:{paginationCount:1,paginationItems:[{id:"81831",title:"Deep Network Model and Regression Analysis using OLS Method for Predicting Lung Vital Capacity",doi:"10.5772/intechopen.104737",signatures:"Harun Sümbül",slug:"deep-network-model-and-regression-analysis-using-ols-method-for-predicting-lung-vital-capacity",totalDownloads:0,totalCrossrefCites:null,totalDimensionsCites:0,authors:null,book:{title:"Decision Science - Recent Advances and Applications",coverURL:"https://cdn.intechopen.com/books/images_new/11604.jpg",subseries:{id:"86",title:"Business and Management"}}}]},subseriesFiltersForOFChapters:[{caption:"Business and Management",value:86,count:1,group:"subseries"}],publishedBooks:{paginationCount:1,paginationItems:[{type:"book",id:"11392",title:"Leadership in a Changing World",subtitle:"A Multidimensional Perspective",coverURL:"https://cdn.intechopen.com/books/images_new/11392.jpg",slug:"leadership-in-a-changing-world-a-multidimensional-perspective",publishedDate:"May 11th 2022",editedByType:"Edited by",bookSignature:"Muhammad Mohiuddin, Bilal Khalid, Md. Samim Al Azad and Slimane Ed-dafali",hash:"86a6d33cf601587e591064ce92effc02",volumeInSeries:1,fullTitle:"Leadership in a Changing World - A Multidimensional Perspective",editors:[{id:"418514",title:"Dr.",name:"Muhammad",middleName:null,surname:"Mohiuddin",slug:"muhammad-mohiuddin",fullName:"Muhammad Mohiuddin",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038UqSfQAK/Profile_Picture_2022-05-13T10:39:03.jpg",institutionString:null,institution:{name:"Université Laval",institutionURL:null,country:{name:"Canada"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null}]},subseriesFiltersForPublishedBooks:[{group:"subseries",caption:"Business and Management",value:86,count:1}],publicationYearFilters:[{group:"publicationYear",caption:"2022",value:2022,count:1}],authors:{paginationCount:249,paginationItems:[{id:"274452",title:"Dr.",name:"Yousif",middleName:"Mohamed",surname:"Abdallah",slug:"yousif-abdallah",fullName:"Yousif Abdallah",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/274452/images/8324_n.jpg",biography:"I certainly enjoyed my experience in Radiotherapy and Nuclear Medicine, particularly it has been in different institutions and hospitals with different Medical Cultures and allocated resources. Radiotherapy and Nuclear Medicine Technology has always been my aspiration and my life. As years passed I accumulated a tremendous amount of skills and knowledge in Radiotherapy and Nuclear Medicine, Conventional Radiology, Radiation Protection, Bioinformatics Technology, PACS, Image processing, clinically and lecturing that will enable me to provide a valuable service to the community as a Researcher and Consultant in this field. My method of translating this into day to day in clinical practice is non-exhaustible and my habit of exchanging knowledge and expertise with others in those fields is the code and secret of success.",institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"313277",title:"Dr.",name:"Bartłomiej",middleName:null,surname:"Płaczek",slug:"bartlomiej-placzek",fullName:"Bartłomiej Płaczek",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/313277/images/system/313277.jpg",biography:"Bartłomiej Płaczek, MSc (2002), Ph.D. (2005), Habilitation (2016), is a professor at the University of Silesia, Institute of Computer Science, Poland, and an expert from the National Centre for Research and Development. His research interests include sensor networks, smart sensors, intelligent systems, and image processing with applications in healthcare and medicine. He is the author or co-author of more than seventy papers in peer-reviewed journals and conferences as well as the co-author of several books. He serves as a reviewer for many scientific journals, international conferences, and research foundations. Since 2010, Dr. Placzek has been a reviewer of grants and projects (including EU projects) in the field of information technologies.",institutionString:"University of Silesia",institution:{name:"University of Silesia",country:{name:"Poland"}}},{id:"35000",title:"Prof.",name:"Ulrich H.P",middleName:"H.P.",surname:"Fischer",slug:"ulrich-h.p-fischer",fullName:"Ulrich H.P Fischer",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/35000/images/3052_n.jpg",biography:"Academic and Professional Background\nUlrich H. P. has Diploma and PhD degrees in Physics from the Free University Berlin, Germany. He has been working on research positions in the Heinrich-Hertz-Institute in Germany. Several international research projects has been performed with European partners from France, Netherlands, Norway and the UK. He is currently Professor of Communications Systems at the Harz University of Applied Sciences, Germany.\n\nPublications and Publishing\nHe has edited one book, a special interest book about ‘Optoelectronic Packaging’ (VDE, Berlin, Germany), and has published over 100 papers and is owner of several international patents for WDM over POF key elements.\n\nKey Research and Consulting Interests\nUlrich’s research activity has always been related to Spectroscopy and Optical Communications Technology. Specific current interests include the validation of complex instruments, and the application of VR technology to the development and testing of measurement systems. He has been reviewer for several publications of the Optical Society of America\\'s including Photonics Technology Letters and Applied Optics.\n\nPersonal Interests\nThese include motor cycling in a very relaxed manner and performing martial arts.",institutionString:null,institution:{name:"Charité",country:{name:"Germany"}}},{id:"341622",title:"Ph.D.",name:"Eduardo",middleName:null,surname:"Rojas Alvarez",slug:"eduardo-rojas-alvarez",fullName:"Eduardo Rojas Alvarez",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/341622/images/15892_n.jpg",biography:null,institutionString:null,institution:{name:"University of Cuenca",country:{name:"Ecuador"}}},{id:"215610",title:"Prof.",name:"Muhammad",middleName:null,surname:"Sarfraz",slug:"muhammad-sarfraz",fullName:"Muhammad Sarfraz",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/215610/images/system/215610.jpeg",biography:"Muhammad Sarfraz is a professor in the Department of Information Science, Kuwait University, Kuwait. His research interests include optimization, computer graphics, computer vision, image processing, machine learning, pattern recognition, soft computing, data science, and intelligent systems. Prof. Sarfraz has been a keynote/invited speaker at various platforms around the globe. He has advised/supervised more than 110 students for their MSc and Ph.D. theses. He has published more than 400 publications as books, journal articles, and conference papers. He has authored and/or edited around seventy books. Prof. Sarfraz is a member of various professional societies. He is a chair and member of international advisory committees and organizing committees of numerous international conferences. He is also an editor and editor in chief for various international journals.",institutionString:"Kuwait University",institution:{name:"Kuwait University",country:{name:"Kuwait"}}},{id:"32650",title:"Prof.",name:"Lukas",middleName:"Willem",surname:"Snyman",slug:"lukas-snyman",fullName:"Lukas Snyman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/32650/images/4136_n.jpg",biography:"Lukas Willem Snyman received his basic education at primary and high schools in South Africa, Eastern Cape. He enrolled at today's Nelson Metropolitan University and graduated from this university with a BSc in Physics and Mathematics, B.Sc Honors in Physics, MSc in Semiconductor Physics, and a Ph.D. in Semiconductor Physics in 1987. After his studies, he chose an academic career and devoted his energy to the teaching of physics to first, second, and third-year students. After positions as a lecturer at the University of Port Elizabeth, he accepted a position as Associate Professor at the University of Pretoria, South Africa.\r\n\r\nIn 1992, he motivates the concept of 'television and computer-based education” as means to reach large student numbers with only the best of teaching expertise and publishes an article on the concept in the SA Journal of Higher Education of 1993 (and later in 2003). The University of Pretoria subsequently approved a series of test projects on the concept with outreach to Mamelodi and Eerste Rust in 1993. In 1994, the University established a 'Unit for Telematic Education ' as a support section for multiple faculties at the University of Pretoria. In subsequent years, the concept of 'telematic education” subsequently becomes well established in academic circles in South Africa, grew in popularity, and is adopted by many universities and colleges throughout South Africa as a medium of enhancing education and training, as a method to reaching out to far out communities, and as a means to enhance study from the home environment.\r\n\r\nProfessor Snyman in subsequent years pursued research in semiconductor physics, semiconductor devices, microelectronics, and optoelectronics.\r\n\r\nIn 2000 he joined the TUT as a full professor. Here served for a period as head of the Department of Electronic Engineering. Here he makes contributions to solar energy development, microwave and optoelectronic device development, silicon photonics, as well as contributions to new mobile telecommunication systems and network planning in SA.\r\n\r\nCurrently, he teaches electronics and telecommunications at the TUT to audiences ranging from first-year students to Ph.D. level.\r\n\r\nFor his research in the field of 'Silicon Photonics” since 1990, he has published (as author and co-author) about thirty internationally reviewed articles in scientific journals, contributed to more than forty international conferences, about 25 South African provisional patents (as inventor and co-inventor), 8 PCT international patent applications until now. Of these, two USA patents applications, two European Patents, two Korean patents, and ten SA patents have been granted. A further 4 USA patents, 5 European patents, 3 Korean patents, 3 Chinese patents, and 3 Japanese patents are currently under consideration.\r\n\r\nRecently he has also published an extensive scholarly chapter in an internet open access book on 'Integrating Microphotonic Systems and MOEMS into standard Silicon CMOS Integrated circuitry”.\r\n\r\nFurthermore, Professor Snyman recently steered a new initiative at the TUT by introducing a 'Laboratory for Innovative Electronic Systems ' at the Department of Electrical Engineering. The model of this laboratory or center is to primarily combine outputs as achieved by high-level research with lower-level system development and entrepreneurship in a technical university environment. Students are allocated to projects at different levels with PhDs and Master students allocated to the generation of new knowledge and new technologies, while students at the diploma and Baccalaureus level are allocated to electronic systems development with a direct and a near application for application in industry or the commercial and public sectors in South Africa.\r\n\r\nProfessor Snyman received the WIRSAM Award of 1983 and the WIRSAM Award in 1985 in South Africa for best research papers by a young scientist at two international conferences on electron microscopy in South Africa. He subsequently received the SA Microelectronics Award for the best dissertation emanating from studies executed at a South African university in the field of Physics and Microelectronics in South Africa in 1987. In October of 2011, Professor Snyman received the prestigious Institutional Award for 'Innovator of the Year” for 2010 at the Tshwane University of Technology, South Africa. This award was based on the number of patents recognized and granted by local and international institutions as well as for his contributions concerning innovation at the TUT.",institutionString:null,institution:{name:"University of South Africa",country:{name:"South Africa"}}},{id:"317279",title:"Mr.",name:"Ali",middleName:"Usama",surname:"Syed",slug:"ali-syed",fullName:"Ali Syed",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/317279/images/16024_n.png",biography:"A creative, talented, and innovative young professional who is dedicated, well organized, and capable research fellow with two years of experience in graduate-level research, published in engineering journals and book, with related expertise in Bio-robotics, equally passionate about the aesthetics of the mechanical and electronic system, obtained expertise in the use of MS Office, MATLAB, SolidWorks, LabVIEW, Proteus, Fusion 360, having a grasp on python, C++ and assembly language, possess proven ability in acquiring research grants, previous appointments with social and educational societies with experience in administration, current affiliations with IEEE and Web of Science, a confident presenter at conferences and teacher in classrooms, able to explain complex information to audiences of all levels.",institutionString:null,institution:{name:"Air University",country:{name:"Pakistan"}}},{id:"75526",title:"Ph.D.",name:"Zihni Onur",middleName:null,surname:"Uygun",slug:"zihni-onur-uygun",fullName:"Zihni Onur Uygun",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/75526/images/12_n.jpg",biography:"My undergraduate education and my Master of Science educations at Ege University and at Çanakkale Onsekiz Mart University have given me a firm foundation in Biochemistry, Analytical Chemistry, Biosensors, Bioelectronics, Physical Chemistry and Medicine. After obtaining my degree as a MSc in analytical chemistry, I started working as a research assistant in Ege University Medical Faculty in 2014. In parallel, I enrolled to the MSc program at the Department of Medical Biochemistry at Ege University to gain deeper knowledge on medical and biochemical sciences as well as clinical chemistry in 2014. In my PhD I deeply researched on biosensors and bioelectronics and finished in 2020. Now I have eleven SCI-Expanded Index published papers, 6 international book chapters, referee assignments for different SCIE journals, one international patent pending, several international awards, projects and bursaries. In parallel to my research assistant position at Ege University Medical Faculty, Department of Medical Biochemistry, in April 2016, I also founded a Start-Up Company (Denosens Biotechnology LTD) by the support of The Scientific and Technological Research Council of Turkey. Currently, I am also working as a CEO in Denosens Biotechnology. The main purposes of the company, which carries out R&D as a research center, are to develop new generation biosensors and sensors for both point-of-care diagnostics; such as glucose, lactate, cholesterol and cancer biomarker detections. My specific experimental and instrumental skills are Biochemistry, Biosensor, Analytical Chemistry, Electrochemistry, Mobile phone based point-of-care diagnostic device, POCTs and Patient interface designs, HPLC, Tandem Mass Spectrometry, Spectrophotometry, ELISA.",institutionString:null,institution:{name:"Ege University",country:{name:"Turkey"}}},{id:"246502",title:"Dr.",name:"Jaya T.",middleName:"T",surname:"Varkey",slug:"jaya-t.-varkey",fullName:"Jaya T. Varkey",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/246502/images/11160_n.jpg",biography:"Jaya T. Varkey, PhD, graduated with a degree in Chemistry from Cochin University of Science and Technology, Kerala, India. She obtained a PhD in Chemistry from the School of Chemical Sciences, Mahatma Gandhi University, Kerala, India, and completed a post-doctoral fellowship at the University of Minnesota, USA. She is a research guide at Mahatma Gandhi University and Associate Professor in Chemistry, St. Teresa’s College, Kochi, Kerala, India.\nDr. Varkey received a National Young Scientist award from the Indian Science Congress (1995), a UGC Research award (2016–2018), an Indian National Science Academy (INSA) Visiting Scientist award (2018–2019), and a Best Innovative Faculty award from the All India Association for Christian Higher Education (AIACHE) (2019). She Hashas received the Sr. Mary Cecil prize for best research paper three times. She was also awarded a start-up to develop a tea bag water filter. \nDr. Varkey has published two international books and twenty-seven international journal publications. She is an editorial board member for five international journals.",institutionString:"St. Teresa’s College",institution:null},{id:"250668",title:"Dr.",name:"Ali",middleName:null,surname:"Nabipour Chakoli",slug:"ali-nabipour-chakoli",fullName:"Ali Nabipour Chakoli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/250668/images/system/250668.jpg",biography:"Academic Qualification:\r\n•\tPhD in Materials Physics and Chemistry, From: Sep. 2006, to: Sep. 2010, School of Materials Science and Engineering, Harbin Institute of Technology, Thesis: Structure and Shape Memory Effect of Functionalized MWCNTs/poly (L-lactide-co-ε-caprolactone) Nanocomposites. Supervisor: Prof. Wei Cai,\r\n•\tM.Sc in Applied Physics, From: 1996, to: 1998, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Determination of Boron in Micro alloy Steels with solid state nuclear track detectors by neutron induced auto radiography, Supervisors: Dr. M. Hosseini Ashrafi and Dr. A. Hosseini.\r\n•\tB.Sc. in Applied Physics, From: 1991, to: 1996, Faculty of Physics & Nuclear Science, Amirkabir Uni. of Technology, Tehran, Iran, Thesis: Design of shielding for Am-Be neutron sources for In Vivo neutron activation analysis, Supervisor: Dr. M. Hosseini Ashrafi.\r\n\r\nResearch Experiences:\r\n1.\tNanomaterials, Carbon Nanotubes, Graphene: Synthesis, Functionalization and Characterization,\r\n2.\tMWCNTs/Polymer Composites: Fabrication and Characterization, \r\n3.\tShape Memory Polymers, Biodegradable Polymers, ORC, Collagen,\r\n4.\tMaterials Analysis and Characterizations: TEM, SEM, XPS, FT-IR, Raman, DSC, DMA, TGA, XRD, GPC, Fluoroscopy, \r\n5.\tInteraction of Radiation with Mater, Nuclear Safety and Security, NDT(RT),\r\n6.\tRadiation Detectors, Calibration (SSDL),\r\n7.\tCompleted IAEA e-learning Courses:\r\nNuclear Security (15 Modules),\r\nNuclear Safety:\r\nTSA 2: Regulatory Protection in Occupational Exposure,\r\nTips & Tricks: Radiation Protection in Radiography,\r\nSafety and Quality in Radiotherapy,\r\nCourse on Sealed Radioactive Sources,\r\nCourse on Fundamentals of Environmental Remediation,\r\nCourse on Planning for Environmental Remediation,\r\nKnowledge Management Orientation Course,\r\nFood Irradiation - Technology, Applications and Good Practices,\r\nEmployment:\r\nFrom 2010 to now: Academic staff, Nuclear Science and Technology Research Institute, Kargar Shomali, Tehran, Iran, P.O. Box: 14395-836.\r\nFrom 1997 to 2006: Expert of Materials Analysis and Characterization. Research Center of Agriculture and Medicine. Rajaeeshahr, Karaj, Iran, P. O. Box: 31585-498.",institutionString:"Atomic Energy Organization of Iran",institution:{name:"Atomic Energy Organization of Iran",country:{name:"Iran"}}},{id:"248279",title:"Dr.",name:"Monika",middleName:"Elzbieta",surname:"Machoy",slug:"monika-machoy",fullName:"Monika Machoy",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/248279/images/system/248279.jpeg",biography:"Monika Elżbieta Machoy, MD, graduated with distinction from the Faculty of Medicine and Dentistry at the Pomeranian Medical University in 2009, defended her PhD thesis with summa cum laude in 2016 and is currently employed as a researcher at the Department of Orthodontics of the Pomeranian Medical University. She expanded her professional knowledge during a one-year scholarship program at the Ernst Moritz Arndt University in Greifswald, Germany and during a three-year internship at the Technical University in Dresden, Germany. She has been a speaker at numerous orthodontic conferences, among others, American Association of Orthodontics, European Orthodontic Symposium and numerous conferences of the Polish Orthodontic Society. She conducts research focusing on the effect of orthodontic treatment on dental and periodontal tissues and the causes of pain in orthodontic patients.",institutionString:"Pomeranian Medical University",institution:{name:"Pomeranian Medical University",country:{name:"Poland"}}},{id:"252743",title:"Prof.",name:"Aswini",middleName:"Kumar",surname:"Kar",slug:"aswini-kar",fullName:"Aswini Kar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/252743/images/10381_n.jpg",biography:"uploaded in cv",institutionString:null,institution:{name:"KIIT University",country:{name:"India"}}},{id:"204256",title:"Dr.",name:"Anil",middleName:"Kumar",surname:"Kumar Sahu",slug:"anil-kumar-sahu",fullName:"Anil Kumar Sahu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/204256/images/14201_n.jpg",biography:"I have nearly 11 years of research and teaching experience. I have done my master degree from University Institute of Pharmacy, Pt. Ravi Shankar Shukla University, Raipur, Chhattisgarh India. I have published 16 review and research articles in international and national journals and published 4 chapters in IntechOpen, the world’s leading publisher of Open access books. I have presented many papers at national and international conferences. I have received research award from Indian Drug Manufacturers Association in year 2015. My research interest extends from novel lymphatic drug delivery systems, oral delivery system for herbal bioactive to formulation optimization.",institutionString:null,institution:{name:"Chhattisgarh Swami Vivekanand Technical University",country:{name:"India"}}},{id:"253468",title:"Dr.",name:"Mariusz",middleName:null,surname:"Marzec",slug:"mariusz-marzec",fullName:"Mariusz Marzec",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/253468/images/system/253468.png",biography:"An assistant professor at Department of Biomedical Computer Systems, at Institute of Computer Science, Silesian University in Katowice. Scientific interests: computer analysis and processing of images, biomedical images, databases and programming languages. He is an author and co-author of scientific publications covering analysis and processing of biomedical images and development of database systems.",institutionString:"University of Silesia",institution:null},{id:"212432",title:"Prof.",name:"Hadi",middleName:null,surname:"Mohammadi",slug:"hadi-mohammadi",fullName:"Hadi Mohammadi",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/212432/images/system/212432.jpeg",biography:"Dr. Hadi Mohammadi is a biomedical engineer with hands-on experience in the design and development of many engineering structures and medical devices through various projects that he has been involved in over the past twenty years. Dr. Mohammadi received his BSc. and MSc. degrees in Mechanical Engineering from Sharif University of Technology, Tehran, Iran, and his PhD. degree in Biomedical Engineering (biomaterials) from the University of Western Ontario. He was a postdoctoral trainee for almost four years at University of Calgary and Harvard Medical School. He is an industry innovator having created the technology to produce lifelike synthetic platforms that can be used for the simulation of almost all cardiovascular reconstructive surgeries. He’s been heavily involved in the design and development of cardiovascular devices and technology for the past 10 years. He is currently an Assistant Professor with the University of British Colombia, Canada.",institutionString:"University of British Columbia",institution:{name:"University of British Columbia",country:{name:"Canada"}}},{id:"254463",title:"Prof.",name:"Haisheng",middleName:null,surname:"Yang",slug:"haisheng-yang",fullName:"Haisheng Yang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/254463/images/system/254463.jpeg",biography:"Haisheng Yang, Ph.D., Professor and Director of the Department of Biomedical Engineering, College of Life Science and Bioengineering, Beijing University of Technology. He received his Ph.D. degree in Mechanics/Biomechanics from Harbin Institute of Technology (jointly with University of California, Berkeley). Afterwards, he worked as a Postdoctoral Research Associate in the Purdue Musculoskeletal Biology and Mechanics Lab at the Department of Basic Medical Sciences, Purdue University, USA. He also conducted research in the Research Centre of Shriners Hospitals for Children-Canada at McGill University, Canada. Dr. Yang has over 10 years research experience in orthopaedic biomechanics and mechanobiology of bone adaptation and regeneration. He earned an award from Beijing Overseas Talents Aggregation program in 2017 and serves as Beijing Distinguished Professor.",institutionString:"Beijing University of Technology",institution:null},{id:"255757",title:"Dr.",name:"Igor",middleName:"Victorovich",surname:"Lakhno",slug:"igor-lakhno",fullName:"Igor Lakhno",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/255757/images/system/255757.jpg",biography:"Lakhno Igor Victorovich was born in 1971 in Kharkiv (Ukraine). \nMD – 1994, Kharkiv National Medical Univesity.\nOb&Gyn; – 1997, master courses in Kharkiv Medical Academy of Postgraduate Education.\nPhD – 1999, Kharkiv National Medical Univesity.\nDSc – 2019, PL Shupik National Academy of Postgraduate Education \nLakhno Igor has been graduated from an international training courses on reproductive medicine and family planning held in Debrecen University (Hungary) in 1997. Since 1998 Lakhno Igor has worked as an associate professor of the department of obstetrics and gynecology of VN Karazin National University and an associate professor of the perinatology, obstetrics and gynecology department of Kharkiv Medical Academy of Postgraduate Education. Since June 2019 he’s a professor of the department of obstetrics and gynecology of VN Karazin National University and a professor of the perinatology, obstetrics and gynecology department of Kharkiv Medical Academy of Postgraduate Education . He’s an author of about 200 printed works and there are 17 of them in Scopus or Web of Science databases. Lakhno Igor is a rewiever of Journal of Obstetrics and Gynaecology (Taylor and Francis), Informatics in Medicine Unlocked (Elsevier), The Journal of Obstetrics and Gynecology Research (Wiley), Endocrine, Metabolic & Immune Disorders-Drug Targets (Bentham Open), The Open Biomedical Engineering Journal (Bentham Open), etc. He’s defended a dissertation for DSc degree \\'Pre-eclampsia: prediction, prevention and treatment”. Lakhno Igor has participated as a speaker in several international conferences and congresses (International Conference on Biological Oscillations April 10th-14th 2016, Lancaster, UK, The 9th conference of the European Study Group on Cardiovascular Oscillations). His main scientific interests: obstetrics, women’s health, fetal medicine, cardiovascular medicine.",institutionString:"V.N. Karazin Kharkiv National University",institution:{name:"Kharkiv Medical Academy of Postgraduate Education",country:{name:"Ukraine"}}},{id:"89721",title:"Dr.",name:"Mehmet",middleName:"Cuneyt",surname:"Ozmen",slug:"mehmet-ozmen",fullName:"Mehmet Ozmen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/89721/images/7289_n.jpg",biography:null,institutionString:null,institution:{name:"Gazi University",country:{name:"Turkey"}}},{id:"243698",title:"M.D.",name:"Xiaogang",middleName:null,surname:"Wang",slug:"xiaogang-wang",fullName:"Xiaogang Wang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/243698/images/system/243698.png",biography:"Dr. Xiaogang Wang, a faculty member of Shanxi Eye Hospital specializing in the treatment of cataract and retinal disease and a tutor for postgraduate students of Shanxi Medical University, worked in the COOL Lab as an international visiting scholar under the supervision of Dr. David Huang and Yali Jia from October 2012 through November 2013. Dr. Wang earned an MD from Shanxi Medical University and a Ph.D. from Shanghai Jiao Tong University. Dr. Wang was awarded two research project grants focused on multimodal optical coherence tomography imaging and deep learning in cataract and retinal disease, from the National Natural Science Foundation of China. He has published around 30 peer-reviewed journal papers and four book chapters and co-edited one book.",institutionString:"Shanxi Eye Hospital",institution:{name:"Shanxi Eye Hospital",country:{name:"China"}}},{id:"242893",title:"Ph.D. Student",name:"Joaquim",middleName:null,surname:"De Moura",slug:"joaquim-de-moura",fullName:"Joaquim De Moura",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/242893/images/7133_n.jpg",biography:"Joaquim de Moura received his degree in Computer Engineering in 2014 from the University of A Coruña (Spain). In 2016, he received his M.Sc degree in Computer Engineering from the same university. He is currently pursuing his Ph.D degree in Computer Science in a collaborative project between ophthalmology centers in Galicia and the University of A Coruña. His research interests include computer vision, machine learning algorithms and analysis and medical imaging processing of various kinds.",institutionString:null,institution:{name:"University of A Coruña",country:{name:"Spain"}}},{id:"267434",title:"Dr.",name:"Rohit",middleName:null,surname:"Raja",slug:"rohit-raja",fullName:"Rohit Raja",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRZkkQAG/Profile_Picture_2022-05-09T12:55:18.jpg",biography:null,institutionString:null,institution:null},{id:"294334",title:"B.Sc.",name:"Marc",middleName:null,surname:"Bruggeman",slug:"marc-bruggeman",fullName:"Marc Bruggeman",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/294334/images/8242_n.jpg",biography:"Chemical engineer graduate, with a passion for material science and specific interest in polymers - their near infinite applications intrigue me. \n\nI plan to continue my scientific career in the field of polymeric biomaterials as I am fascinated by intelligent, bioactive and biomimetic materials for use in both consumer and medical applications.",institutionString:null,institution:null},{id:"244950",title:"Dr.",name:"Salvatore",middleName:null,surname:"Di Lauro",slug:"salvatore-di-lauro",fullName:"Salvatore Di Lauro",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0030O00002bSF1HQAW/ProfilePicture%202021-12-20%2014%3A54%3A14.482",biography:"Name:\n\tSALVATORE DI LAURO\nAddress:\n\tHospital Clínico Universitario Valladolid\nAvda Ramón y Cajal 3\n47005, Valladolid\nSpain\nPhone number: \nFax\nE-mail:\n\t+34 983420000 ext 292\n+34 983420084\nsadilauro@live.it\nDate and place of Birth:\nID Number\nMedical Licence \nLanguages\t09-05-1985. Villaricca (Italy)\n\nY1281863H\n474707061\nItalian (native language)\nSpanish (read, written, spoken)\nEnglish (read, written, spoken)\nPortuguese (read, spoken)\nFrench (read)\n\t\t\nCurrent position (title and company)\tDate (Year)\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. Private practise.\t2017-today\n\n2019-today\n\t\n\t\nEducation (High school, university and postgraduate training > 3 months)\tDate (Year)\nDegree in Medicine and Surgery. University of Neaples 'Federico II”\nResident in Opthalmology. Hospital Clinico Universitario Valladolid\nMaster in Vitreo-Retina. IOBA. University of Valladolid\nFellow of the European Board of Ophthalmology. Paris\nMaster in Research in Ophthalmology. University of Valladolid\t2003-2009\n2012-2016\n2016-2017\n2016\n2012-2013\n\t\nEmployments (company and positions)\tDate (Year)\nResident in Ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl.\nFellow in Vitreo-Retina. IOBA. University of Valladolid\nVitreo-Retinal consultant in ophthalmology. Hospital Clinico Universitario Valladolid. Sacyl. National Health System.\nVitreo-Retinal consultant in ophthalmology. Instituto Oftalmologico Recoletas. Red Hospitalaria Recoletas. \n\t2012-2016\n2016-2017\n2017-today\n\n2019-Today\n\n\n\t\nClinical Research Experience (tasks and role)\tDate (Year)\nAssociated investigator\n\n' FIS PI20/00740: DESARROLLO DE UNA CALCULADORA DE RIESGO DE\nAPARICION DE RETINOPATIA DIABETICA BASADA EN TECNICAS DE IMAGEN MULTIMODAL EN PACIENTES DIABETICOS TIPO 1. Grant by: Ministerio de Ciencia e Innovacion \n\n' (BIO/VA23/14) Estudio clínico multicéntrico y prospectivo para validar dos\nbiomarcadores ubicados en los genes p53 y MDM2 en la predicción de los resultados funcionales de la cirugía del desprendimiento de retina regmatógeno. Grant by: Gerencia Regional de Salud de la Junta de Castilla y León.\n' Estudio multicéntrico, aleatorizado, con enmascaramiento doble, en 2 grupos\nparalelos y de 52 semanas de duración para comparar la eficacia, seguridad e inmunogenicidad de SOK583A1 respecto a Eylea® en pacientes con degeneración macular neovascular asociada a la edad' (CSOK583A12301; N.EUDRA: 2019-004838-41; FASE III). Grant by Hexal AG\n\n' Estudio de fase III, aleatorizado, doble ciego, con grupos paralelos, multicéntrico para comparar la eficacia y la seguridad de QL1205 frente a Lucentis® en pacientes con degeneración macular neovascular asociada a la edad. (EUDRACT: 2018-004486-13). Grant by Qilu Pharmaceutical Co\n\n' Estudio NEUTON: Ensayo clinico en fase IV para evaluar la eficacia de aflibercept en pacientes Naive con Edema MacUlar secundario a Oclusion de Vena CenTral de la Retina (OVCR) en regimen de tratamientO iNdividualizado Treat and Extend (TAE)”, (2014-000975-21). Grant by Fundacion Retinaplus\n\n' Evaluación de la seguridad y bioactividad de anillos de tensión capsular en conejo. Proyecto Procusens. Grant by AJL, S.A.\n\n'Estudio epidemiológico, prospectivo, multicéntrico y abierto\\npara valorar la frecuencia de la conjuntivitis adenovírica diagnosticada mediante el test AdenoPlus®\\nTest en pacientes enfermos de conjuntivitis aguda”\\n. National, multicenter study. Grant by: NICOX.\n\nEuropean multicentric trial: 'Evaluation of clinical outcomes following the use of Systane Hydration in patients with dry eye”. Study Phase 4. Grant by: Alcon Labs'\n\nVLPs Injection and Activation in a Rabbit Model of Uveal Melanoma. Grant by Aura Bioscience\n\nUpdating and characterization of a rabbit model of uveal melanoma. Grant by Aura Bioscience\n\nEnsayo clínico en fase IV para evaluar las variantes genéticas de la vía del VEGF como biomarcadores de eficacia del tratamiento con aflibercept en pacientes con degeneración macular asociada a la edad (DMAE) neovascular. Estudio BIOIMAGE. IMO-AFLI-2013-01\n\nEstudio In-Eye:Ensayo clínico en fase IV, abierto, aleatorizado, de 2 brazos,\nmulticçentrico y de 12 meses de duración, para evaluar la eficacia y seguridad de un régimen de PRN flexible individualizado de 'esperar y extender' versus un régimen PRN según criterios de estabilización mediante evaluaciones mensuales de inyecciones intravítreas de ranibizumab 0,5 mg en pacientes naive con neovascularización coriodea secunaria a la degeneración macular relacionada con la edad. CP: CRFB002AES03T\n\nTREND: Estudio Fase IIIb multicéntrico, randomizado, de 12 meses de\nseguimiento con evaluador de la agudeza visual enmascarado, para evaluar la eficacia y la seguridad de ranibizumab 0.5mg en un régimen de tratar y extender comparado con un régimen mensual, en pacientes con degeneración macular neovascular asociada a la edad. CP: CRFB002A2411 Código Eudra CT:\n2013-002626-23\n\n\n\nPublications\t\n\n2021\n\n\n\n\n2015\n\n\n\n\n2021\n\n\n\n\n\n2021\n\n\n\n\n2015\n\n\n\n\n2015\n\n\n2014\n\n\n\n\n2015-16\n\n\n\n2015\n\n\n2014\n\n\n2014\n\n\n\n\n2014\n\n\n\n\n\n\n\n2014\n\nJose Carlos Pastor; Jimena Rojas; Salvador Pastor-Idoate; Salvatore Di Lauro; Lucia Gonzalez-Buendia; Santiago Delgado-Tirado. Proliferative vitreoretinopathy: A new concept of disease pathogenesis and practical\nconsequences. Progress in Retinal and Eye Research. 51, pp. 125 - 155. 03/2016. DOI: 10.1016/j.preteyeres.2015.07.005\n\n\nLabrador-Velandia S; Alonso-Alonso ML; Di Lauro S; García-Gutierrez MT; Srivastava GK; Pastor JC; Fernandez-Bueno I. Mesenchymal stem cells provide paracrine neuroprotective resources that delay degeneration of co-cultured organotypic neuroretinal cultures.Experimental Eye Research. 185, 17/05/2019. DOI: 10.1016/j.exer.2019.05.011\n\nSalvatore Di Lauro; Maria Teresa Garcia Gutierrez; Ivan Fernandez Bueno. Quantification of pigment epithelium-derived factor (PEDF) in an ex vivo coculture of retinal pigment epithelium cells and neuroretina.\nJournal of Allbiosolution. 2019. ISSN 2605-3535\n\nSonia Labrador Velandia; Salvatore Di Lauro; Alonso-Alonso ML; Tabera Bartolomé S; Srivastava GK; Pastor JC; Fernandez-Bueno I. Biocompatibility of intravitreal injection of human mesenchymal stem cells in immunocompetent rabbits. Graefe's archive for clinical and experimental ophthalmology. 256 - 1, pp. 125 - 134. 01/2018. DOI: 10.1007/s00417-017-3842-3\n\n\nSalvatore Di Lauro, David Rodriguez-Crespo, Manuel J Gayoso, Maria T Garcia-Gutierrez, J Carlos Pastor, Girish K Srivastava, Ivan Fernandez-Bueno. A novel coculture model of porcine central neuroretina explants and retinal pigment epithelium cells. Molecular Vision. 2016 - 22, pp. 243 - 253. 01/2016.\n\nSalvatore Di Lauro. Classifications for Proliferative Vitreoretinopathy ({PVR}): An Analysis of Their Use in Publications over the Last 15 Years. Journal of Ophthalmology. 2016, pp. 1 - 6. 01/2016. DOI: 10.1155/2016/7807596\n\nSalvatore Di Lauro; Rosa Maria Coco; Rosa Maria Sanabria; Enrique Rodriguez de la Rua; Jose Carlos Pastor. Loss of Visual Acuity after Successful Surgery for Macula-On Rhegmatogenous Retinal Detachment in a Prospective Multicentre Study. Journal of Ophthalmology. 2015:821864, 2015. DOI: 10.1155/2015/821864\n\nIvan Fernandez-Bueno; Salvatore Di Lauro; Ivan Alvarez; Jose Carlos Lopez; Maria Teresa Garcia-Gutierrez; Itziar Fernandez; Eva Larra; Jose Carlos Pastor. Safety and Biocompatibility of a New High-Density Polyethylene-Based\nSpherical Integrated Porous Orbital Implant: An Experimental Study in Rabbits. Journal of Ophthalmology. 2015:904096, 2015. DOI: 10.1155/2015/904096\n\nPastor JC; Pastor-Idoate S; Rodríguez-Hernandez I; Rojas J; Fernandez I; Gonzalez-Buendia L; Di Lauro S; Gonzalez-Sarmiento R. Genetics of PVR and RD. Ophthalmologica. 232 - Suppl 1, pp. 28 - 29. 2014\n\nRodriguez-Crespo D; Di Lauro S; Singh AK; Garcia-Gutierrez MT; Garrosa M; Pastor JC; Fernandez-Bueno I; Srivastava GK. Triple-layered mixed co-culture model of RPE cells with neuroretina for evaluating the neuroprotective effects of adipose-MSCs. Cell Tissue Res. 358 - 3, pp. 705 - 716. 2014.\nDOI: 10.1007/s00441-014-1987-5\n\nCarlo De Werra; Salvatore Condurro; Salvatore Tramontano; Mario Perone; Ivana Donzelli; Salvatore Di Lauro; Massimo Di Giuseppe; Rosa Di Micco; Annalisa Pascariello; Antonio Pastore; Giorgio Diamantis; Giuseppe Galloro. Hydatid disease of the liver: thirty years of surgical experience.Chirurgia italiana. 59 - 5, pp. 611 - 636.\n(Italia): 2007. ISSN 0009-4773\n\nChapters in books\n\t\n' Salvador Pastor Idoate; Salvatore Di Lauro; Jose Carlos Pastor Jimeno. PVR: Pathogenesis, Histopathology and Classification. Proliferative Vitreoretinopathy with Small Gauge Vitrectomy. Springer, 2018. ISBN 978-3-319-78445-8\nDOI: 10.1007/978-3-319-78446-5_2. \n\n' Salvatore Di Lauro; Maria Isabel Lopez Galvez. Quistes vítreos en una mujer joven. Problemas diagnósticos en patología retinocoroidea. Sociedad Española de Retina-Vitreo. 2018.\n\n' Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor Jimeno. iOCT in PVR management. OCT Applications in Opthalmology. pp. 1 - 8. INTECH, 2018. DOI: 10.5772/intechopen.78774.\n\n' Rosa Coco Martin; Salvatore Di Lauro; Salvador Pastor Idoate; Jose Carlos Pastor. amponadores, manipuladores y tinciones en la cirugía del traumatismo ocular.Trauma Ocular. Ponencia de la SEO 2018..\n\n' LOPEZ GALVEZ; DI LAURO; CRESPO. OCT angiografia y complicaciones retinianas de la diabetes. PONENCIA SEO 2021, CAPITULO 20. (España): 2021.\n\n' Múltiples desprendimientos neurosensoriales bilaterales en paciente joven. Enfermedades Degenerativas De Retina Y Coroides. SERV 04/2016. \n' González-Buendía L; Di Lauro S; Pastor-Idoate S; Pastor Jimeno JC. Vitreorretinopatía proliferante (VRP) e inflamación: LA INFLAMACIÓN in «INMUNOMODULADORES Y ANTIINFLAMATORIOS: MÁS ALLÁ DE LOS CORTICOIDES. RELACION DE PONENCIAS DE LA SOCIEDAD ESPAÑOLA DE OFTALMOLOGIA. 10/2014.",institutionString:null,institution:null},{id:"265335",title:"Mr.",name:"Stefan",middleName:"Radnev",surname:"Stefanov",slug:"stefan-stefanov",fullName:"Stefan Stefanov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/265335/images/7562_n.jpg",biography:null,institutionString:null,institution:null},{id:"318905",title:"Prof.",name:"Elvis",middleName:"Kwason",surname:"Tiburu",slug:"elvis-tiburu",fullName:"Elvis Tiburu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Ghana",country:{name:"Ghana"}}},{id:"336193",title:"Dr.",name:"Abdullah",middleName:null,surname:"Alamoudi",slug:"abdullah-alamoudi",fullName:"Abdullah Alamoudi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Majmaah University",country:{name:"Saudi Arabia"}}},{id:"318657",title:"MSc.",name:"Isabell",middleName:null,surname:"Steuding",slug:"isabell-steuding",fullName:"Isabell Steuding",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}},{id:"318656",title:"BSc.",name:"Peter",middleName:null,surname:"Kußmann",slug:"peter-kussmann",fullName:"Peter Kußmann",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Harz University of Applied Sciences",country:{name:"Germany"}}},{id:"338222",title:"Mrs.",name:"María José",middleName:null,surname:"Lucía Mudas",slug:"maria-jose-lucia-mudas",fullName:"María José Lucía Mudas",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Carlos III University of Madrid",country:{name:"Spain"}}},{id:"147824",title:"Mr.",name:"Pablo",middleName:null,surname:"Revuelta Sanz",slug:"pablo-revuelta-sanz",fullName:"Pablo Revuelta Sanz",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Carlos III University of Madrid",country:{name:"Spain"}}}]}},subseries:{item:{id:"12",type:"subseries",title:"Human Physiology",keywords:"Anatomy, Cells, Organs, Systems, Homeostasis, Functions",scope:"Human physiology is the scientific exploration of the various functions (physical, biochemical, and mechanical properties) of humans, their organs, and their constituent cells. The endocrine and nervous systems play important roles in maintaining homeostasis in the human body. Integration, which is the biological basis of physiology, is achieved through communication between the many overlapping functions of the human body's systems, which takes place through electrical and chemical means. Much of the basis of our knowledge of human physiology has been provided by animal experiments. Because of the close relationship between structure and function, studies in human physiology and anatomy seek to understand the mechanisms that help the human body function. 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His interest later turned to the molecular mechanism and attenuating strategy of sarcopenia (age-related muscle atrophy). His opinion is to attenuate sarcopenia by improving autophagic defects using nutrient- and pharmaceutical-based treatments.",institutionString:null,institution:{name:"Tokyo Institute of Technology",institutionURL:null,country:{name:"Japan"}}},editorTwo:null,editorThree:{id:"331519",title:"Dr.",name:"Kotomi",middleName:null,surname:"Sakai",slug:"kotomi-sakai",fullName:"Kotomi Sakai",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000031QtFXQA0/Profile_Picture_1637053227318",biography:"Senior researcher Kotomi Sakai, Ph.D., MPH, works at the Research Organization of Science and Technology in Ritsumeikan University. She is a researcher in the geriatric rehabilitation and public health field. She received Ph.D. from Nihon University and MPH from St.Luke’s International University. 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In recent years, the application of chemistry to biological molecules has gained significant interest in medicinal and pharmacological studies. This topic will be devoted to understanding the interplay between biomolecules and chemical compounds, their structure and function, and their potential applications in related fields. Being a part of the biochemistry discipline, the ideas and concepts that have emerged from Chemical Biology have affected other related areas. 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Behind these definitions are hidden all the aspects of normal and pathological functioning of all processes that the topic ‘Metabolism’ will cover within the Biochemistry Series. 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Thus proteomics, an area of research that detects all protein forms expressed in an organism, including splice isoforms and post-translational modifications, is more suitable than genomics for a comprehensive understanding of the biochemical processes that govern life. The most common proteomics applications are currently in the clinical field for the identification, in a variety of biological matrices, of biomarkers for diagnosis and therapeutic intervention of disorders. From the comparison of proteomic profiles of control and disease or different physiological states, which may emerge, changes in protein expression can provide new insights into the roles played by some proteins in human pathologies. Understanding how proteins function and interact with each other is another goal of proteomics that makes this approach even more intriguing. Specialized technology and expertise are required to assess the proteome of any biological sample. Currently, proteomics relies mainly on mass spectrometry (MS) combined with electrophoretic (1 or 2-DE-MS) and/or chromatographic techniques (LC-MS/MS). MS is an excellent tool that has gained popularity in proteomics because of its ability to gather a complex body of information such as cataloging protein expression, identifying protein modification sites, and defining protein interactions. 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