Comparison of Various Expression Systems for Producing Recombinant Proteins
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He holds two PhDs in Mathematics and Prognostics from the Lebanese University and Aix-Marseille University. His research interests are in the field of mathematics.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"248271",title:"Dr.",name:"Abdo",middleName:null,surname:"Abou Jaoudé",slug:"abdo-abou-jaoude",fullName:"Abdo Abou Jaoudé",profilePictureURL:"https://mts.intechopen.com/storage/users/248271/images/system/248271.jpg",biography:"Abdo Abou Jaoudé has been teaching for many years and has a passion for researching and teaching mathematics. He is currently an Associate Professor of Mathematics and Statistics at Notre Dame University-Louaizé (NDU), Lebanon. He holds a BSc and an MSc in Computer Science from NDU, and three PhDs in Applied Mathematics, Computer Science, and Applied Statistics and Probability, all from Bircham International University through a distance learning program. He also holds two PhDs in Mathematics and Prognostics from the Lebanese University, Lebanon, and Aix-Marseille University, France. Dr. Abou Jaoudé's broad research interests are in the field of applied mathematics. He has published twenty-three international journal articles and six contributions to conference proceedings, in addition to seven books on prognostics, pure and applied mathematics, and computer science.",institutionString:"Notre Dame University - Louaize",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"4",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Notre Dame University – Louaize",institutionURL:null,country:{name:"Lebanon"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"15",title:"Mathematics",slug:"mathematics"}],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. 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A vast majority of developing countries cannot afford the high costs of medical treatments resulted from the existing methods. Hence, we need to produce not only the new drugs but also the cheaper versions of the present samples in the market. Molecular farming can offer efficient solutions for the current growing need for the biomedicines [1]. Plants provide an inexpensive and simple system for the production of valuable recombinant proteins on large scale, and compared to the other production systems, they have numerous advantages in terms of economy, safety, and applicability. Though using transgenic plants has entailed some sorts of limitations and concerns, the optimization has been operated for solving the existing problems. Normally, the production of pharmaceutical proteins has been largely concentrated by the technology of molecular farming in plants, also plants can be used for the production of food supplements, biopolymers, industrial enzymes, and proteins in the investigations (avidin, β-glucuronidase, etc.). Prior production systems, including bacteria, microbial eukaryotes (yeasts, double-stranded fungi), animal cells, and transgenic animals, as a result of their limitations, were replaced by transgenic plants. The primary recombinant pharmaceutical proteins, extracted from the plants (hormones of human growth), and the first recombinant antibodies were generated from transgenic plants, respectively, in 1986 and 1989 [2, 3]. In 1997, the first recombinant protein, avidin (egg protein) was produced in a transgenic maize for industrial uses [4]. These applications proved that plants can be converted into bioreactors to produce a wide range of recombinant proteins. Many years had already passed when it was proved that plants were even able to produce several complexes of functional mammal proteins with the pharmacological functions, such as human serum proteins, growth regulators, antibodies, vaccines, hormones, cytokines, and enzymes [5]. An increasing request for the biomedicines was aligned with the high costs and inefficiency of existing production systems [6] including yeasts, bacteria, animal cells [7], and transgenic animals [8].
The aim of this study is to review the technologies of molecular farming, limitations and advantages of plant systems, challenges, bio-security, public acceptance of molecular farming.
Plant molecular farming depending on the production of transgenic plants has been operated by two general methods as the following:
a. Stable nuclear transformation: Stable nuclear transformation refers to the integration of genes or nominated foreign genes into the nuclear genome of plants, which results in the change of genetic structures and consequent expression of transgenes after integration with the host genomes. The largest amount of recombinant proteins has been produced by one of the most common methods of stable nuclear transformation. A method exploited for aggregating proteins in dried beans of maize culminates in a long-term storage in the beans at the room temperature without decomposition of proteins [9]. In addition, it has a considerable potential for producing crops like cereals that actually grow everywhere. However, a long production cycle for some crops and their potential collisions with natural species or food products have restricted the wide acceptance of this method [10].
b. Stable plastid transformation: Plastid transformation offers a remarkable solution in comparison to that of nuclear transformation since it has numerous advantages including preventing transgene escape through amphimixis (because plastids are inherited through the maternal tissue in the majority of species.) and absence of chloroplasts in pollen and consequent improbability of their transfer, which reduces environmental concerns [11, 12]. The transformed transgenic plants with homoplasmic chloroplasts (all chloroplasts carry transgenes) were selected after several generations of plant regeneration from bombarded leaf explants. Selection was conducted on a medium containing spectinomycin or combined with streptomycin. The researchers [13, 14] have already extracted a human pharmaceutical protein, more than 3% to 6%, from the total soluble proteins in the chloroplasts of tobacco. Recently, Oey et al. [75] reported a very high level (70% of an entire soluble protein) for a protein antibiotic with the chloroplast system, which, till today, has been the highest concentration of recombinant proteins. Despite this, the great potential of plastid transformation has some functional limitations. Although this technology has been developed in other species such as tomatoes, lettuce, soy, and eggs [15, 16], in the current situation, chloroplast transformation only in tobacco is practically possible, but unfortunately this plant is inedible and full of poisonous alkaloids; in addition, long lasting storage in refrigerators will bring about changes in protein stability [9].
c. Plant cell suspension culture: This method involves the removal of cell walls and gene transfer to the obtained protoplasts and suspension culture. The purification system and its downstream processing are cheaper and easier [17]. In addition, the use of suspension culture can decrease heterogeneity in proteins and sugar (N-glycans) regarding the uniformity of the type and size of cells [5]. Furthermore, as a fast system there is no need for the production of transgenic plants; however, the cell lines can be produced after a few months [18, 19]. Some samples of plant suspension cultures can be used for producing biomedicines, including vaccines of Newcastle disease virus of chicks approved by the Center for Veterinary Biology and recombinant human glucocerebrosidase for treating diabetes (www.protalix.com) [19]. Though this method is cheaper, safer, and easier in comparison to the other methods of genetic manipulation, cell suspension has not yet been suggested as an optimal production choice of production in plant systems. This is due to a belief that the ultimate products and their usability are constrained by reducing the level of recombinant proteins during the stationary phase because of the enhanced proteolytic activity [20].
A transient production may be the fastest system for plant molecular farming [21]. Nowadays, these are the systems routinely applied for verifying expression constructs during a few weeks for a significant amount of proteins [22]. The given systems include the following methods:
Agrobacterium transformation method: Infiltration of recombinant agrobacterium suspension into tobacco leaf tissue is achieved without stable gene transfer, which facilitates the transfer of T-DNA to a very high percentage of cells, where the transgenes are expressed at a high level without a stable transfer of genes. Presently, this method has been very efficient for the production of clinical biomedicines with a fast expansion [22-24].
Viral infection methods: The viral infection method depends on the capability of plant viruses, such as tobacco mosaic virus and X potato virus, which functions as a vector to convey foreign genes into plant genomes without fusing with the genome of that plant [25].
Magnifection system: Expression systems based on viral vectors and agrobacterium methods suffer from some constraints for the co-expression of two or more polypeptides required for the production of
Comparison of different expression systems (see Table 1) reveals the advantages of plants in comparison with other expression systems as follows:
The healthiness of derived products (plants cannot be the host of human pathogens and bacterial toxins).
Capability of post-translational processing (respecting the features of eukaryotic cells).
The possibility of using breeding methods and sexual crosses to obtain active recombinant multi-chain proteins (therefore, there is the possibility of producing antibodies without application of a double transformation).
Reducing the costs of production (plants can produce biological materials by the use of carbon dioxide, solar energy, and inorganic materials. Moreover, the scale of production can be manipulated regarding scalability).
Reducing the costs of storage and transportation of recombinant proteins (when they are produced in dry textures like grains).
Removing the purification step (when the plant tissues containing recombinant protein are edible) [1].
\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t\t\n\t\t\t\t | \n\t\t
Production cost | \n\t\t\tAverage | \n\t\t\tHigh | \n\t\t\tLow | \n\t\t\tLow | \n\t\t
Post-translational modifications | \n\t\t\tNo | \n\t\t\tYes | \n\t\t\tYes | \n\t\t\tYes | \n\t\t
Function | \n\t\t\tHigh | \n\t\t\tAverage | \n\t\t\tHigh | \n\t\t\tHigh | \n\t\t
Protein stability | \n\t\t\tYes | \n\t\t\tYes | \n\t\t\tYes in seeds | \n\t\t\tYes | \n\t\t
Comparison of Various Expression Systems for Producing Recombinant Proteins
To optimize the expression of transcripts, a widely used strategy is the use of building promoters, such as cauliflower mosaic virus 35S RNA promoter and maize 1-ubiquitin promoter, respectively, suitable for spilt-cotyledons and single-cotyledons [27]. Tissue-specific and organ-specific promoters are used for stimulating the expression of transgenes (antigen vaccine HBsAgM, single-chain variable fragment Maureen G4, and Human interferon-α) in some tissues or organs, such as tubers, seeds, and fruits [28, 29]. The given specific expression of tissues prevents the accumulation of recombinant proteins in vegetative organs, which can have a negative impact on plant growth; for example,
Expression constructs can be designed for guaranteeing the efficiency of translation and the sustainability of transcripts. As an instance, the removal of 5\' untranslated region and natural ´3 for foreign genes and introducing the leader sequence of tobacco mosaic virus RNA, RUB13 rice polyubiquitin gene, alfalfa mosaic virus, or tobacco viruses in the expressions, all, individually, have shown a significant increase in the level of transgene expressions [32, 33].
In addition to the leader sequences, expression cassettes can be designed with the AU-rich sequences in 3\' untranslated regions, which may change or be removed as the editing sites for ensuring the stability of transcript. It is also proved that every organism shows codon usage deviations that may be the subject of importance for adapting the coding sequence of heterologous genes for the host gene to optimize the efficiency of translation. In this regard, the site of initial translation from heterologous protein to pair with Kozak consensus sequence, with the application of GCTTCCTCC sequence, started after codon or ACC, or ACA had been changed before that. It is better to unscientifically estimate codon changes rather than their real amount considering the changes in the expression level of transgenes in similar systems and the use of similar structures. To this end, an increase of codon combinations of (A/G)(a/c)(a/g)AUG and (A/g)(u/C)(g/C)AUG for the optimal operation of translation was, respectively, reported in Arabidopsis and rice. The given change in transgene expression could be due to the position effect, number of transgene copies, or gene silencing.
Regarding the effect of position, expression cassettes can be designed to have nuclear matrix attachment regions for ensuring the transgene insertion in proper sites for stimulating transcription factors for promoters. Furthermore, the problem of position effect can be prohibited by targeting the transgene to plastids. To optimize the production of single-cotyledon transgene, the strategies that include the use of specific genetic elements containing cAMP response elements for a simultaneous transfer with transgene in T-DNA are used. In addition, one new technology, including the structure of an artificial autonomous mini-chromosome, can genetically materialize excellent possibilities with several advantages, namely genetic stability due to the absence of gene silencing and position effect.
To optimize the stability of recombinant proteins, known as the most important limiting factor for the function of molecular farming [34], the targeting of proteins into certain intracellular parts is demanded. The intracellular targeting not only increases protein stability but also determines the processing type of dependent protein. This can be applied for the optimization of isolation and procedures of purification by the fusion proteins and targets with high affinity [27]. Targeting of proteins can be done by the following pathways and organelles,
The intracellular parts, like
Cathepsin D
To protect proteins from cytosolic degradation, these proteins can be targeted by fusion to a C-terminal tail without a forced passing through the lumen of the endoplasmic reticulum to the membrane surface [39]. To enhance the ease of purification, proteins can be fused to
The proteins, like in glycosylation, that do not need post-translation modifications for their activity, can be targeted to chloroplast since post-translation modifications are not conducted in these organelles [40].
Targeting for accumulation in endoplasmic reticulum is accomplished by two methods: one is adding SEKDEL endoplasmic reticulum signals to the end of C-protein, and the other is using fused N or C signals with y-zein. Endoplasmic reticulum is an oxidizing environment with high amounts of
Glycosylation refers to the covalent binding of sugars to proteins in order to increase close-packing, biological activity, solubility, and biological functionality [5]. Glycosylation takes place in plants in the secretory pathway of endoplasmic reticulum and golgi apparatus. The glycosylation patterns of plants and animals differ in the composition of N-glycans; plants add residues of α (1, 3) fucose and β-(1,2) xylose to N-glycans of their protein, but animals add residues of (1 and 6) fucose, glucose, and sialic acid to N-glycans. These differences can be problematic for humans when medical animal proteins extracted from plants are used (Krupp et al., 2003); consequently, a correct human N-glycosylation demands a plant engineering. A number of strategies for changing the pattern of N-glycosylation in plants have been elaborated as following [71]:
The use of purified enzymes of β-(1,4) galactosyltransferase and Sialyltransferase for making glass transitions in the recombinant proteins derived from plants.
Co-expression of β-(1,4) galactosyltransferase human enzyme with the target transgene in transgenic plants.
Prohibiting the activity of fucosyltransferase and xylyltransferase enzymes.
Targeting pharmaceutical proteins to the endoplasmic reticulum in order to avoid the addition of protective N-glycans.
Major economical factors in appointing an appropriate host include the total biomass yield, storage characteristics, ease of transport, value of recombinant proteins, maintenance costs, its availability for workers, required area, duration of production cycle, cost of subsequent products, and edibility [27, 34]. In addition to the economical analysis, a sufficient host should be appropriate in terms of transformation and regeneration [34, 72]. In addition to the high potential of tobacco for transformation and regeneration, it has the majority of the aforementioned economic benefits [27, 41, 42]. However, tobacco (except the
The importance of intracellular localization of proteins is due to the functional consequences of proteins. Therefore, the problem of intracellular localization of amino acid sequences has been the subject of great attention in the community of bioinformatics. Thus, various methods, like searching for targeted signals, have been presented with respect to a prediction that various proteins are produced in different intercellular segments [51].
Plants are able to produce those bacterial and viral recombinant antigens that preserve the capability of making the structures Type IV similar to those witnessed in mammalian systems, and the post-translational modifications are operated to maintain the biological activity of proteins. The most important issue is vaccine production in the edible tissues of transgenic plants, which is a very safe and effective method in vaccination.
The biomedicines produced in plants are as follows:
Antigens for the production of edible vaccines: Antigens, used for generating an immune response resulting in immunity against diseases in human proteins, are expressed from different pathogens in plants. Those vaccines derived from plants have been so far induced immunity against rabies virus, hepatitis B, rotavirus, HIV, and other pathogens.
Monoclonal antibodies: Widespread application of antibodies has lead to the study of new methods in order to strengthen efficiency and reduce the cost of producing antibodies. Among the studied methods, using transgenic plants as bioreactors are known as the most efficient one. While designing therapeutic antibodies in the production of recombinant expression systems, the apprehension of the functioning mechanisms of antibodies is essential. Although the primary function of antibodies is actualized by binding to antigens, it does not act as a protective performance. Some antibodies have a direct neutralizing impact, for instance blocking the bacteria or the active sites of the pathogenic factors such as enzymes. The antibodies produced in plants incorporate Immunoglobulin G (IgG) and Immunoglobulin A (IgA), IgA and IgG shimmer molecules, IgG and IgA secreted molecules, Single-Chain variable fragment, fragment antigen-binding, and second variable of heavy and light chains [52-54].
Pharmaceutical proteins: Some samples of biomedicines recently expressed in plants include
Non-pharmaceutical proteins derived from plants or industrial proteins belong mainly to the enzymes that include avidin, trypsin, aprotinin, β-
Molecular farming and metabolic engineering make the production of new high-tech products possible. There is a driving force backing molecular farming that makes its costs much less than traditional farming.
Recovery usually includes the process and breakdown of plant tissues, protein extraction, solid-liquid separation, and protein concentration while purification encompasses safety protection, liquid-liquid extraction, membrane filtration, chromatography, etc. The processing of leaves requires a particular attention; leaves should be processed immediately after the harvest or frozen to prevent protein degradation by proteases, whereas seeds can be stored for a long period of time due to the less probability of destruction of recombinant proteins expressed in seeds. Using the secretory systems of cells can also be beneficial since disintegrating plant cells throughout recovery is not required; thus, the release of phenolic compounds can be avoided while the recombinant proteins can be unstable in culture mediums. Another way of facilitating the recovery of proteins is utilizing continuous labels. Protein labels must be removed after purification so that the structure of purified protein can change into its natural position. The technology of oleosin fusion, through which the gene sequence of recombinant proteins is fused to the sequence of a special internal oil protein called oleosin in safflower and canola, is separated after the digestion of internal protease following protein purification [1].
The costs of subsequent processing of the recombinant proteins derived from plants have been estimated about 80% of the total production costs [60, 61]. This is why so much attention has been paid at sufficient strategies for reducing the costs to the least amount. The application of watery textures like tomatoes as a production system has been expanded because of their potential for reducing the costs via the ease of extracting from their textures in comparison with those of dry tissues like grains [34, 62]. In addition, tomatoes are highly regarded as a reputed host crop in terms of its bio-safety because these plants grow in greenhouses without worrying about the preservation of transgenic plants.
Nowadays, oil bodies of oilseed agricultural products, like the seeds of safflower and mustard, are being exploited by the application of oleosin fusion technology developed by SemBioSys Genetics in order to facilitate the purification of recombinant proteins and reduction of subsequent costs (http://www.sembiosys.com/). The strategies including targeting of recombinant proteins for the seeds of oilseed agricultural products as an oleosin fusion facilitate the extraction of fused proteins from oil bodies and the release of the recombinant proteins from their fusion partner; one example can be the accumulation and purification of biologically active human insulin, apolipoprotein
There are several recombinant proteins derived from plants that were the basic idea of edible vaccines, directly eaten as fruits (tomatoes and bananas) and vegetables (lettuce and carrots); accordingly, no processing costs will be demanded by the elimination of processing, [66]. Bananas, as a fruit host in agricultural products, have particularly attracted lots of customers for the production of edible vaccines, especially for developing countries. This has been widely developed in such countries because of long distance transports and cooling requirements [42]. Apart from the mentioned advantages, high digestibility and palatability of bananas have won a wide public acceptance for the vaccination of children [67, 68]. The sufficiency of potatoes, eaten in raw or low processed forms, for edible vaccines has resulted in their wide production. Potatoes, like seeds, have the advantage of production stability due to a special molecular environment allocated in glands [69].
The risks of transgenic plants are divided into two categories: one category directly affects humans and the other endangers environment and other organisms. The attack of immune system can disable these medicines and lead to the stimuli for the allergic reactions, some of which have been elaborated as follows:
There are some concerns in terms of environmental pollution about the entrance of transgenes into the food chain, which requires a sound management and supervision.
The other concern refers to the grain transformations using agrobacterium since grains are important crops in the production of pharmaceutical protein.
The reactions of immune system can disable the medicines produced in plants and be the stimuli for allergic reactions [70].
The development stages and subsequent commercialization of the products is the subject of consideration in the second phase of clinical trials. A number of small biotechnology companies have aimed to commercialize the antibodies produced in plants. It has been estimated that the increasing annual need for secretory IgA will be 13%, and a rate of $25 billion was predicted as the annual income of producing IgA in crops. While there have been great advances in the field of biomedicine production in plants on large scales, fundamental studies are demanded to pave the way for the commercialization of these products. The present problems include the difficulty of low yield of protein, the possibility of harmful effects on the function/performance of proteins due to the differences in glycosylation patterns, and the severe potential impact of expressing plants of biomedicine plants on the environment (e.g., concerns upon genetic limitations) [74].
The aim of molecular farming is to produce large quantities of active and secure pharmaceutical proteins with lower prices. With the scientific advances in the field of bio-technology, nowadays, gene transfer methods in plants have considerably developed. These transgenic plants in comparison with other microbial and animal expression systems have various advantages in terms of easy production, cost, safety, etc. for producing pharmaceutical biomolecules. So far, lots of valuable pharmaceutical proteins and antibodies have been produced by the help of this method, which remarkably has helped the treatment of patients especially in developing countries where the production and preservation costs of such medicines cannot be afforded. However, there are some disputes, such as public acceptance, transgene escape and biosecurity, clinical and commercialization investigations of products, etc., which has made it a challenging area, but it is hoped that in near future molecular farming will witness great achievements with the researchers and scholars\' efforts.
We are grateful to the Azarbaijan Shahid Madani University, especially the Vice president for research since financial assistance for some research in this field.
Stable isotope labeling with amino acids in cell culture (SILAC) is a polypeptide-labeling technology developed by the Thermo Fisher company of the United States in 2002 [1]. Heavy isotopes (13C or 15N) and light isotopes (12C or 14N) are used to label two essential amino acids (L-lysine and L-arginine) that are contained in a cell-cultured medium, respectively. After the cells were cultured with essential amino acids for 6–10 generations, all proteins were labeled with heavy isotopes or light isotopes. The cellular proteins stimulated by different treatment factors are analyzed by mass spectrometry (MS) to obtain the qualitative and quantitative proteome data [2]. SILAC generally allows heavy and light isotopes-labeled sample cells at the early stage of the experimental workflow, so the variability caused by the sample handling process was minimized [3]. SILAC was widely used in quantitative proteomics to study pathogenesis, drug target, protein modification and dynamics, protein-molecule interaction, and screen special functional proteins [4]. SILAC showed outstanding performance for quantification and dynamics of phosphosites in colorectal cancer with the treatment of the epidermal growth factor receptor (EGFR)-blocking antibody cetuximab, rendering it the effective method for cellular signaling study in cell culture models [5]. In terms of identification of protein-molecule interaction, SILAC combined with various affinity purifications of protein experimental setups could be used to distinguish specific complexes from nonspecific ones [6]. One study performed SILAC to overcome the most challenging problem in defining specific partners in protein complexes. The cells containing an affinity tagged protein were cultured in a light isotopic medium, while wild-type cells were grown in a heavy isotopic medium. The results of MS showed that specific partners appear as isotopically light [7]. SILAC also offers numerous opportunities to discover potential biomarkers and therapeutic targets for some drugs [8]. SILAC in combination with other developed approaches made SILAC more popular; for example, these SILAC labels in pulse or pulse-chase scenarios could be used to measure macromolecular dynamics on time scales of several hours [9]. An MS-based approach combining dynamic-SILAC labeling with isobaric mass tagging was well used to understand protein degradation and synthesis in cellular systems [10]. SILAC provided an effective scheme to comprehensively and systematically qualify and quantify complex mammalian cell proteome, which would promote progress in the medical field.
Ivermectin, marketed in 1981, was commonly used as a broad-spectrum antiparasitic compound. It was approved to treat onchocerciasis (150–200 μg kg−1 body weight), scabies (200 μg kg−1 body weight), lymphatic filariasis (150–200 μg kg−1 body weight), demodicosis (200 μg kg−1 body weight), strongyloidiasis (200 μg kg−1 body weight), pediculosis (400 μg kg−1 body weight), and filariasis (due to Mansonella ozzardi, 6 mg as a single dose) [11].
Because ivermectin mainly targets chloride-dependent channels (γ-aminobutyric acid and glutamate), its safety could be fine in higher animals. In humans, especially the blood-brain barrier can reduce ivermectin delivery to the central nervous system [12]. The safety of ivermectin has been proved with clinical studies on children, infants, and pregnant women. A study including 170 infants and children with the treatment of oral ivermectin (mean dose = 223 μg kg−1) showed good tolerance, and only seven subjects occurred mild adverse events [13]. A study including 893 pregnant women with the oral treatment of ivermectin also showed good tolerance, and no patients were reported to generate serious events (stillbirths, neonatal death, low birth weight, spontaneous abortions, preterm births, and congenital anomalies) [14]. Those studies proved that ivermectin was safe enough to be used in human diseases, but there was still insufficient evidence to prove no adverse side effects. The highest ivermectin dose was 200 μg/kg, which was approved by FDA. However, some patients without serious events have used 10 times more than the FDA-approved dose [15]. All those made ivermectin more likely to achieve success in clinical application. In recent days, studies found that ivermectin was effective in a completely new range of diseases, such as neurological disorders, antiviral (e.g., dengue, HIV, and encephalitis), antibacterial (e.g., Buruli ulcer and tuberculosis), anticancer (melanoma, lymphoid leukemia, lung cancer, glioblastoma, and breast cancer) [11]. The functions and mechanisms of ivermectin on anticancer generated interest and excitement in the scientific community. Ivermectin suppresses breast cancer by disrupting cellular signaling in the process and activating cytostatic autophagy through mediating PAK1 expression [16]. Ivermectin showed a synergistic effect with the chemotherapy agents by increasing cell death in leukemia cells. Some researchers, who aimed at overcoming cancer, claimed that ivermectin could be rapidly advanced into clinical trials [17]. Further study on molecular network, signaling pathway, and key biological processes of ivermectin would provide more useful information about this multifaceted “wonder” drug.
This chapter describes that SILAC identifies differentially expressed proteins (DEGs) in ivermectin-treated ovarian cancer cells in the following aspects: (i) ovarian cancer cell culture—TOV21G and labeled with heavy and light SILAC reagents; (ii) ivermectin treatment of SILAC-labeled TOV21G cells and protein preparation; (iii) the quality of SILAC-labeled protein samples with 1D SDS-PAGE; (iv) trypsin-digestion of SILAC-labeled proteins; (v) each fraction was subjected to LC-MS/MS analysis; and (vi) bioinformatics analysis (signaling pathway and biological process). SILAC can be a useful and effective method to detect protein alterations and dynamic changes in living cells, and the results would provide scientific data to further clarify molecular mechanisms of ivermectin in ovarian cancer.
The human ovarian cell line (TOV-21G) was cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and 5% CO2 at 37°C. (i) Ovarian cell line used here was TOV-21G, which was obtained from Keibai Academy of Science (Nanjing, China) [18]. (ii) RPMI 1640 was used without glutamine, lysine, and arginine. (iii) FBS was brought from Gibco® Certified Thermo Fisher Scientific. (iv) The growing states of TOV-21G were observed, and the medium was changed in every 2 days.
SILAC “light” or “heavy” labeling growing medium (Thermo Fisher Scientific, US) was used to culture TOV-21G cells for 10 passages, to ensure a high level of stable isotope replacing original amino acids [8]. (i) “Light” labeled amino acids: 50 mg L-arginine HCl (Arg0), 50 mg L-lysine HCl (Lys0). “Heavy” labeled amino acids: 50 mg L-arginine-13C6,15N4 HCl (Arg10), 50 mg L-lysine-13C6,15N2 HCl (Lys8). (ii) A total of 1 L RPMI 1640 without glutamine, lysine, and arginine. (iii) For SILAC experiments, 50 mg Arg0 and 50 mg Lys0 (“light” labeling reagent) were added into 500 mL RPMI 1640 medium to form SILAC “light” labeling growing medium. A total of 50 mg Arg10 and 50 mg Lys8 (“heavy” labeling reagent) were added to 500 mL RPMI 1640 medium to form SILAC “heavy” labeling growing medium. (iv) The growing states of TOV-21G were observed, and the medium was changed in every 2 days. TOV-21G cells were cultured and passaged for 10 generations in 10-cm culture flasks.
When SILAC-labeled TOV21G cells achieved 80% cell density in 10-cm culture flasks, 20 μM ivermectin was added to TOV-21G cells in SILAC “heavy” labeling growing medium, and 0.1% DMSO was added to TOV-21G cells in SILAC “light” labeling growing medium for 24 h [19]. (i) Ivermectin (C48H74O14, purity ≥ 95%): The drug was brought from Solarbio (http://www.solarbio.com/goods-3911.html). (ii) TOV-21G cells were counted, and 8000 cells/well were seeded in 96-well plates. Ivermectin was added into each well in different drug concentrations (0 μM, 10 μM, 20 μM, 30 μM, 40 μM, and 50 μM) for 24 h. CCK8 (10 μL) was added into each well for 1 h to measure absorbance values (OD) at a wavelength of 450 nm. Lethal concentration 50 (IC50) was calculated according to OD values of each well in different ivermectin concentrations. (iii) Ivermectin treatment group: 20 μM ivermectin was added to TOV-21G cells in SILAC “heavy” labeling growing medium for 24 h. Control group: 0.1% DMSO was added to TOV-21G cells in SILAC “light” labeling growing medium for 24 h.
Ivermectin treatment and 0.1% DMSO treatment TOV-21G cells were collected and lysed by protein isolation buffer, respectively. (i) TOV-21G cells collection: A total of 500 μL trypsin was added to each 10-cm culture flasks for several minutes and collected with centrifugation (800 × g, 5 min). TOV-21G cells were washed with ice-cold phosphate buffer solution (PBS) for three times. (ii) Protein isolation buffer: 2 mM thiourea, 4% CHAPS (3-[(3-cholamidopropyl)-dimethylammonio] -1-propane), 7 M urea, 100 mM dithiothreitol (DTT), and 2% ampholyte. (iii) TOV-21G cells lysis: A total of 200 μL protein isolation buffer was added to each 10-cm culture flasks for 30 min (ice-cold) and then oscillated with five vortex cycles. (iv) SILAC-labeled protein collection: Protein isolation buffer was centrifuged (13,000 × g, 20 min, 4°C), and the SILAC-labeled protein samples were collected from the supernatants in new tubes. (v) Protein concentrations measurement: Protein concentrations of the SILAC-labeled protein samples were measured with the 2-D quant protein assay kit (Bio-Rad, US).
The “heavy”- and “light”-SILAC-labeled proteins were mixed and loaded onto 1X SDS-PAGE to check the quality. SDS-PAGE-separated proteins were further analyzed with MS/MS as a preliminary experiment to check the labeling efficiency. (i) The loading sample preparation: According to the 1:1 ratio, the “heavy”- and “light”-SILAC-labeled proteins were mixed in a 5X loading buffer. (ii) Electrophoresis: The mixed SILAC-labeled proteins were loaded onto SDS-PAGE gel (gel concentration: 12.5%) with the amount of 20 μg/lane by constant current (14 mA, 90 min). (iii) Coomassie brilliant blue staining: Prepare Coomassie brilliant blue stain and destain solutions. Filter the stain solution through Whatman 1 filter paper. (iv) MS/MS: Proteins were separated from SDS-PAGE bands, and then were reduced, alkylated, and trypsin-digested. The tryptic peptides were analyzed with MS/MS.
The main reagents and methods included: (i) Reducing agent: 100 mM DTT was added to SILAC-labeled protein sample. (ii) Uranyl acetate (UA) buffer: The UA buffer contained 8 M urea and 0.1 M Tris/HCL. The SILAC-labeled protein sample with DTT was filtered by a 10-kD ultrafiltration centrifuge tube for two times. (iii) Isolation mixture reacted: A total of 100 μL of 0.05 M iodoacetamide was added to the isolation mixture following centrifugalization (14,000 × g, 15 min). A total of 25 mM ammonium bicarbonate (NH4HCO3) was added to the mixture following centrifugalization (14,000 × g, 15 min). (iv) Trypsin buffer: 2 μg trypsin in 40 μL 100-mM NH4HCO3. (v) Tryptic peptide content: A volume of trypsin buffer (40 μL) was added to the mixture from last step and shaken by 600 rpm for 1 min. Enzymatic hydrolysis of the mixture was done for 16–18 h at 37°C. A volume (40 μL) of 25 mM NH4HCO3 was added to the mixture and that mixture was centrifuged (14,000 × g, 15 min); the filtrate was collected.
The instrument and materials are as follows: (i) MS instrument, for example, Q Exactive mass spectrometer (Thermo Fisher Scientific); (ii) Easy nLC system, for example, Proxeon Biosystems (Thermo Fisher Scientific); (iii) Thermo scientific EASY column: Acclaim PepMap, 100 μm × 2 cm, nanoViper, 5 μm-C18; (iv) analytical column: Thermo scientific EASY column (75 μm * 100 mm 3 μm-C18); (v) solvent A: 0.1% formic acid in H2O; and (vi) solvent B: 0.1% formic acid, 84% acetonitrile in H2O.
The main parameters are following [20]: Main search ppm: 6; missed cleavage: 2; MS/MS tolerance ppm: 20; de-isotopic: TRUE; enzyme: trypsin/P; database: uniprot_Homo_sapiens_169753_20190313; fixed modification: carbamidomethyl (C); lables: Lys(8), Arg(10); variable modification: oxidation (M), acetyl (protein N-term) decoy database; pattern: reverse; peptide FDR: 0.01; and protein FDR: 0.01.
Several bioinformatics analyses were used, which are as follows: (i) The enrichment of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway was performed with R package clusterProfiler-KEGG (https://bioconductor.org/packages/release/bioc/html/clusterProfiler.html). (ii) The enrichment of biological processes (BPs) was analyzed with Cytoscape ClueGO. (iii) The level of statistical significance was set as p < 0.05 and adjusted p value < 0.05. For KEGG and BPs enrichment analyses, a Benjamini-Hochberg multiple text was used to adjust p value.
The flow chart of SILAC quantitative proteomics was shown to summarize the overall analysis process for the identification of ivermectin-related proteins (Figure 1).
The flow chart of SILAC quantitative proteomics analysis of ovarian cancer cells treated with and without ivermectin.
In total, 4447 proteins were identified with SILAC quantitative proteomics in human ovarian cancer cells treated with ivermectin. The ratio of “heavy”/“light” labeling samples was obtained, including 97.91% proteins with ratio < 1, and 2.09% proteins with ratio > 1. The MS/MS spectra of tryptic peptides EYQDLLNVK (Figure 2A) and VVQGSLDSLPQAVR (Figure 2B) are taken as examples. For peptide EYQDLLNVK (gene name = NEFM), the excellent b-ion and y-ion series were obtained with high signal-to-noise (S/N) (Figure 2A). For peptide VVQGSLDSLPQAVR (gene name = PKC1), the excellent b-ion and y-ion series were also obtained with high signal-to-noise (S/N) (Figure 2B).
The MS/MS spectra of tryptic peptides with SILAC labeling. (A) MS/MS spectrum of tryptic peptide EYQDLLNVK (gene name = NEFM). (B) MS/MS spectrum of tryptic peptide VVQGSLDSLPQAVR (gene name = PKC1).
The fold-changes of some identified proteins were very striking; for example, those upregulated proteins (ratio > 2), including histone H2A, progranulin, cathepsin Z, SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A-like protein 1, beta-mannosidase, GRAM domain-containing 1C, BMP-2-inducible protein kinase, ribosomal protein L3, ubiquitin-conjugating enzyme E2, PIK3R1, LIM domain-containing protein 1, retinal guanylyl cyclase 1, telomerase-binding protein EST1A, and COX7A2L protein. Some of them have been reported in ovarian cancers. For example, recent studies demonstrated that PI3K/AKT/mTOR and ERK1/2 signaling pathways were involved in this chemoresistance. Progranulin was upregulated in epithelial ovarian cancer cell lines and associated with cisplatin resistance through regulating AKT/mTOR and ERK1/2 signaling pathways [21]. Progranulin is also involved in the process of cartilage development, progression, wound healing, and inflammation in ovarian cancer [22]. Additionally, one study showed that progranulin could directly activate cancer-associated fibroblasts to induce the epithelial-mesenchymal transition process of epithelial ovarian cancer cells [23]. Copy number loss of PIK3R1 most commonly occurs in ovarian cancer, which would activate AKT and p110-independent JAK2/STAT3 signaling and renders ovarian cancer cells vulnerable to AKT inhibitors [24]. CD97 can activate NF-κB-dependent JAK2/STAT3 pathway, consequently playing an important role in migratory, invasive capacity, and drug-resistant in ovarian cancer cells [25].
Some downregulated proteins (ratio < 0.1) were also very striking; for example, anion exchange protein, Rho guanine nucleotide exchange factor 16, dynein assembly factor 1, glucoside xylosyltransferase 1, glutamine synthetase, insulin-like growth factor-binding protein 2, microtubule-associated protein RP/EB family member 3, myelin proteolipid protein, neurochondrin, neurofilament medium polypeptide, phosphoenolpyruvate carboxykinase, ANKUB1, and TASOR 2. Some of them play an important role in the pathogenesis of ovarian cancer. For example, the most prominent effects of insulin-like growth factor-binding protein 2 in ovarian cancer include promoting driving invasion, proliferation, and suppressing apoptosis. The area under the ROC curve of insulin-like growth factor-binding protein 2 in detecting ovarian cancer was 0.815 (95% CI: 0.721–0.910, P < 0.001), further studies are needed to confirm its diagnostic performance at an early stage of ovarian cancer [26].
The glutamine metabolism could be a novel therapeutic target against cisplatin resistance in various cancers. Glutamine synthetase can take part in the reprogramming of glutamine metabolism to induce cisplatin resistance in A2780 ovarian cancer cells [27]. Anion exchanger 2 is a sodium-independent chloride/bicarbonate transporter, which is implicated in the regulation of membrane potential and intracellular potential of hydrogen (pH value). Anion exchanger 2 was highly expressed in ovarian cancer tissues compared to adjacent non-tumor lesions with quantitative proteomics analysis [28]. Those identified proteins in ovarian cancer cells treated with and without ivermectin with SILAC quantitative proteomics discovered reliable and effective biomarkers and drug targets for the anticancer process of ivermectin [8].
In total, 89 statistically significant molecular pathways were enriched based on those 4447 ivermectin-related proteins with KEGG pathway analysis (Table 1).
hsa00010 | Glycolysis/gluconeogenesis | 2.03E−03 |
hsa00020 | Citrate cycle (TCA cycle) | 7.38E−11 |
hsa00030 | Pentose phosphate pathway | 3.51E−04 |
hsa00052 | Galactose metabolism | 6.04E−03 |
hsa00062 | Fatty acid elongation | 1.09E−02 |
hsa00071 | Fatty acid degradation | 1.39E−05 |
hsa00190 | Oxidative phosphorylation | 3.47E−12 |
hsa00230 | Purine metabolism | 1.08E−02 |
hsa00240 | Pyrimidine metabolism | 2.55E−03 |
hsa00270 | Cysteine and methionine metabolism | 8.93E−07 |
hsa00280 | Valine, leucine and isoleucine degradation | 2.06E−08 |
hsa00480 | Glutathione metabolism | 1.05E−03 |
hsa00510 | N-Glycan biosynthesis | 4.31E−03 |
hsa00513 | Various types of N-glycan biosynthesis | 5.15E−03 |
hsa00520 | Amino sugar and nucleotide sugar metabolism | 1.04E−07 |
hsa00620 | Pyruvate metabolism | 1.11E−07 |
hsa00630 | Glyoxylate and dicarboxylate metabolism | 4.09E−03 |
hsa00640 | Propanoate metabolism | 1.16E−05 |
hsa00920 | Sulfur metabolism | 4.45E−03 |
hsa01040 | Biosynthesis of unsaturated fatty acids | 1.09E−02 |
hsa01200 | Carbon metabolism | 3.96E−16 |
hsa01212 | Fatty acid metabolism | 1.74E−08 |
hsa01230 | Biosynthesis of amino acids | 3.09E−08 |
hsa03008 | Ribosome biogenesis in eukaryotes | 3.34E−04 |
hsa03010 | Ribosome | 7.07E−22 |
hsa03013 | RNA transport | 8.04E−20 |
hsa03015 | mRNA surveillance pathway | 4.28E−08 |
hsa03018 | RNA degradation | 2.24E−07 |
hsa03030 | DNA replication | 8.18E−09 |
hsa03040 | Spliceosome | 5.79E−24 |
hsa03050 | Proteasome | 1.50E−14 |
hsa03410 | Base excision repair | 1.22E−02 |
hsa03420 | Nucleotide excision repair | 4.72E−06 |
hsa03430 | Mismatch repair | 4.33E−04 |
hsa04012 | ErbB signaling pathway | 6.27E−03 |
hsa04066 | HIF-1 signaling pathway | 1.31E−05 |
hsa04071 | Sphingolipid signaling pathway | 9.01E−03 |
hsa04110 | Cell cycle | 4.03E−03 |
hsa04120 | Ubiquitin mediated proteolysis | 1.82E−06 |
hsa04130 | SNARE interactions in vesicular transport | 1.18E−04 |
hsa04137 | Mitophagy—animal | 9.07E−03 |
hsa04141 | Protein processing in the endoplasmic reticulum | 1.07E−13 |
hsa04142 | Lysosome | 3.81E−06 |
hsa04144 | Endocytosis | 9.24E−15 |
hsa04145 | Phagosome | 8.66E−07 |
hsa04152 | AMPK signaling pathway | 3.49E−03 |
hsa04210 | Apoptosis | 1.07E−03 |
hsa04213 | Longevity regulating pathway—multiple species | 4.46E−03 |
hsa04216 | Ferroptosis | 4.75E−04 |
hsa04510 | Focal adhesion | 2.57E−05 |
hsa04520 | Adherens junction | 2.04E−05 |
hsa04530 | Tight junction | 6.21E−05 |
hsa04540 | Gap junction | 3.14E−03 |
hsa04611 | Platelet activation | 4.03E−03 |
hsa04666 | Fc gamma R-mediated phagocytosis | 2.41E−03 |
hsa04714 | Thermogenesis | 1.08E−09 |
hsa04720 | Long-term potentiation | 7.22E−03 |
hsa04721 | Synaptic vesicle cycle | 2.29E−04 |
hsa04810 | Regulation of actin cytoskeleton | 4.94E−06 |
hsa04910 | Insulin signaling pathway | 2.90E−04 |
hsa04919 | Thyroid hormone signaling pathway | 5.20E−03 |
hsa04922 | Glucagon signaling pathway | 3.38E−04 |
hsa04931 | Insulin resistance | 1.26E−02 |
hsa04932 | Nonalcoholic fatty liver disease (NAFLD) | 1.44E−07 |
hsa04961 | Endocrine and other factor-regulated calcium reabsorption | 1.83E−03 |
hsa04962 | Vasopressin-regulated water reabsorption | 9.81E−03 |
hsa05010 | Alzheimer disease | 3.09E−07 |
hsa05012 | Parkinson disease | 5.32E−23 |
hsa05014 | Amyotrophic lateral sclerosis (ALS) | 2.55E−03 |
hsa05016 | Huntington disease | 7.93E−13 |
hsa05100 | Bacterial invasion of epithelial cells | 5.95E−10 |
hsa05110 | 6.22E−06 | |
hsa05120 | Epithelial cell signaling in | 1.07E−04 |
hsa05130 | Pathogenic | 6.13E−09 |
hsa05131 | Shigellosis | 2.10E−07 |
hsa05132 | Salmonella infection | 7.10E−14 |
hsa05134 | Legionellosis | 1.05E−03 |
hsa05135 | Yersinia infection | 4.64E−06 |
hsa05163 | Human cytomegalovirus infection | 5.57E−05 |
hsa05165 | Human papillomavirus infection | 4.68E−03 |
hsa05169 | Epstein-Barr virus infection | 2.03E−05 |
hsa05170 | Human immunodeficiency virus 1 infection | 4.59E−08 |
hsa05203 | Viral carcinogenesis | 6.98E−05 |
hsa05205 | Proteoglycans in cancer | 2.44E−05 |
hsa05211 | Renal cell carcinoma | 2.65E−03 |
hsa05212 | Pancreatic cancer | 6.79E−03 |
hsa05220 | Chronic myeloid leukemia | 1.31E−02 |
hsa05230 | Central carbon metabolism in cancer | 1.18E−03 |
hsa03060 | Protein export | 8.73E−05 |
Statistically significant pathways identified with ivermectin-related proteins with KEGG pathway enrichment analysis.
These molecular pathways demonstrated that ivermectin was involved in multiple cancer-related molecular pathways, such as mismatch repair process, ErbB signaling pathway, HIF-1 signaling pathway, cell-cycle regulation, ubiquitin-mediated proteolysis, AMPK signaling pathway, apoptosis, ferroptosis, proteoglycans, and central carbon metabolism in cancer. These molecular pathways also indicated that ivermectin was involved in multiple cancer pathogenesis, such as energy metabolism pathways, immunity-related pathways, stromal element-related pathways, RNA regulation pathways, hormone signaling pathways, and biosynthesis of substances. Different pathways enriched different proteins, whereas some pathways shared the same proteins. These data showed that ivermectin has a complex influence on various signaling pathways. The results were consistent with many studies previously. Ivermectin induced PAK1-mediated cytostatic autophagy both
A total of 61 statistically significant biological processes were enriched based on those 4447 ivermectin-related proteins with GO analysis (Figure 3). These biological processes indicated that ivermectin was involved in multiple cancer-related biological processes, such as negative/positive regulation of canonical Wnt signaling pathway, cysteine-type endopeptidase activity involved in the apoptotic process, innate immune response-activating signal transduction, protein targeting to membrane, T-cell receptor signaling pathway, regulation of protein ubiquitination, activation of protein kinase activity, regulation of transcription by RNA polymerase II, and DNA-binding transcription factor activity. These results were consistent with many studies previously. For example, CTNNB1 (catenin beta 1, IMPβ1) in the biological process of protein polyubiquitination, was responsible for the nuclear entry of cargoes. Ivermectin can impact thermal stability and α-helicity of IMPα and IMPβ1 by binding to the IMPα armadillo repeat domain [35]. CASP3 in the biological process of protein kinase regulator activity is a member of the cysteine-aspartic acid protease (caspase) family. SK-MEL-28 cells were treated with different concentrations of ivermectin (2.5 μM, 5 μM, and 10 μM). Ivermectin enhanced the apoptosis effect by the upregulation of caspase-3 activity [36]. Also, PAK1 in the biological process of protein kinase regulator activity binds to and inhibits the activity of cyclin-cyclin-dependent kinase 2 or -cyclin-dependent kinase 4 complexes, and thus functions as a regulator of cell-cycle progression at G1. Ivermectin inhibited cancer stem cells formation by regulating the binding of PAK1/Stat3 complex and the IL-6 promoter [37]. YAP1 in the biological process of positive regulation of canonical Wnt signaling pathway was involved in the development, growth, repair, and homeostasis of multiple cancers. Ivermectin inhibited YAP1 nuclear expression and nuclear accumulation in gastric cancer cells. Moreover, in xenografts of gastric cancer cells, ivermectin suppressed tumor growth by regulating YAP1 nuclear expression [38]. Those identified proteins in ovarian cancer cells treated with and without ivermectin based on the SILAC method play important roles in multiple cellular signaling pathways and have broad biological activities. Those findings provide basic data for further study of ivermectin in ovarian cancer.
Statistically significant biological processes (BPs) identified with ivermectin-related proteins with GO enrichment analysis.
Stable isotope labeling with amino acids in cell culture (SILAC) was an effective quantitative proteomics method to identify differentially expressed proteins or differentially modified proteins in cultured cells between two different conditions. In this study, ovarian cancer cells TOV-21 under two different conditions were cultured with the “heavy” labeling medium that contained 50 mg L-lysine-2HCl [13C6, 15N2] and 50 mg L-arginine-HCl [13C6, 15N4] in 500 mL RPMI 1640 medium, and the “light” labeling medium that contained 50 mg L-lysine-2HCl [12C6, 14N2] and 50 mg L-arginine-HCl [12C6, 14N4] in 500 mL RPMI 1640 medium for 10 passages, respectively. Then TOV-21G cells with SILAC “heavy” or “light” labeling were treated with or without 20 μM ivermectin for 24 h. The heavy- and light-stable isotope-labeled proteins were equally mixed (1:1), digested with trypsin, and analyzed with LC-MS/MS. A total of 4447 proteins were identified in ivermectin-treated TOV-21G cells relative to controls, and these proteins were significantly enriched in 89 molecular pathways, and 62 biological processes. These findings offer important data to study ivermectin-mediated molecular pathway network changes and discover effective ivermectin-related biomarkers and therapeutic targets for ivermectin treatment of ovarian cancer.
The authors acknowledge the financial supports from the Shandong First Medical University Talent Introduction Funds (to X.Z.), and the Hunan Provincial Hundred Talent Plan (to X.Z.).
We declare that we have no financial and personal relationships with other people or organizations.
N.L. analyzed the data, prepared figures, and wrote the manuscript. X.Z. conceived the concept, designed the manuscript, wrote and critically revised the manuscript, and was responsible for the correspondence work and financial support.
BPs | biological processes |
CTNNB1 | catenin beta 1 |
ICD | immunogenic cell death |
DTT | dithiothreitol |
EGFR | epidermal growth factor receptor |
IC50 | lethal concentration 50 |
KEGG | Kyoto Encyclopedia of Genes and Genomes |
LC | liquid chromatography |
MS | mass spectrometry |
MS/MS | tandem mass spectrometry |
OD | absorbance values |
PBS | phosphate buffer solution |
SILAC | stable isotope labeling with amino acids in cell culture |
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Iron oxide nanoparticles and their nanocomposites have performed excellent in supercapacitor. Iron oxide as negative electrode has extended the working voltage window of a supercapacitor. The main problems associated with iron oxide based electrodes are their poor electrical conductivity and cycle stability. Therefore, a conductive carbon matrix has been added to the iron oxide based electrodes to improve the electrochemical performance. In this chapter, recent progress on iron oxide and its composite with different materials as electrode in supercapacitor is summarized. The various synergistic effects of nanocomposites and compositional engineering to enhance the electrochemical performance of iron oxide are also discussed.",book:{id:"10824",title:"Iron Oxide Nanoparticles",coverURL:"https://cdn.intechopen.com/books/images_new/10824.jpg"},signatures:"Rajan Lakra, Rahul Kumar, Parasanta Kumar Sahoo, Sandeep Kumar and Ankur Soam"},{id:"82030",title:"Magnetite Nanoparticles (Fe3O4) for Radio-Frequency and Microwave Applications",slug:"magnetite-nanoparticles-fe3o4-for-radio-frequency-and-microwave-applications",totalDownloads:7,totalDimensionsCites:0,doi:"10.5772/intechopen.104930",abstract:"The size and shape dependent tunable electromagnetic (EM) properties of magnetite – Fe3O4 nanoparticles makes them an attractive material for various future electronics and biomedical device applications such as tunable attenuators, miniaturized isolators and circulators, RF antennas, EM shielding, and biomedical implants etc. The strategic design of RF devices requires specific dielectric and magnetic properties according to the applications, which in turn depends on the size and shape of the particles. 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Finally, Fe3O4 nanocomposites will be explored by using different synthesis approaches for implementation of RF and microwave applications and we will conclude the chapter with future recommendations.",book:{id:"10824",title:"Iron Oxide Nanoparticles",coverURL:"https://cdn.intechopen.com/books/images_new/10824.jpg"},signatures:"Poonam Lathiya and Jing Wang"},{id:"81878",title:"Recent Progress and Overview of Nanocomposites",slug:"recent-progress-and-overview-of-nanocomposites",totalDownloads:17,totalDimensionsCites:0,doi:"10.5772/intechopen.102469",abstract:"Nanocomposites are versatile materials because of possessing superior properties as compared to their parent materials. Due to their improved electrical, mechanical, thermomechanical, electronic, optoelectronic, thermal, and magnetic properties, these materials are receiving much attention from researchers all over the world. In every field, the focus of the research is to develop such materials which have low weight, superior strength, and enhanced performance as well as cost competitiveness in comparison to existing materials. The nanocomposite materials have been used in the fields of avionics, biomedical, auto industry, sports industry, oil/gas, construction, food industry, agriculture industry, and information technology. 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She is now a lecturer at the University of Witwatersrand, South Africa, and a principal researcher at the Health Economics and Epidemiology Research Office (HE2RO), South Africa. Dr. Moolla holds a Ph.D. in Psychology with her research being focused on mental health and resilience. In her professional work capacity, her research has further expanded into the fields of early childhood development, mental health, the HIV and TB care cascades, as well as COVID. She is also a UNESCO-trained International Bioethics Facilitator.",institutionString:"University of the Witwatersrand",institution:{name:"University of the Witwatersrand",country:{name:"South Africa"}}},{id:"342152",title:"Dr.",name:"Santo",middleName:null,surname:"Grace Umesh",slug:"santo-grace-umesh",fullName:"Santo Grace Umesh",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/342152/images/16311_n.jpg",biography:null,institutionString:null,institution:{name:"SRM Dental College",country:{name:"India"}}},{id:"333647",title:"Dr.",name:"Shreya",middleName:null,surname:"Kishore",slug:"shreya-kishore",fullName:"Shreya Kishore",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/333647/images/14701_n.jpg",biography:"Dr. Shreya Kishore completed her Bachelor in Dental Surgery in Chettinad Dental College and Research Institute, Chennai, and her Master of Dental Surgery (Orthodontics) in Saveetha Dental College, Chennai. She is also Invisalign certified. She’s working as a Senior Lecturer in the Department of Orthodontics, SRM Dental College since November 2019. She is actively involved in teaching orthodontics to the undergraduates and the postgraduates. Her clinical research topics include new orthodontic brackets, fixed appliances and TADs. She’s published 4 articles in well renowned indexed journals and has a published patency of her own. Her private practice is currently limited to orthodontics and works as a consultant in various clinics.",institutionString:null,institution:{name:"SRM Dental College",country:{name:"India"}}},{id:"323731",title:"Prof.",name:"Deepak M.",middleName:"Macchindra",surname:"Vikhe",slug:"deepak-m.-vikhe",fullName:"Deepak M. Vikhe",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/323731/images/13613_n.jpg",biography:"Dr Deepak M.Vikhe .\n\n\t\n\tDr Deepak M.Vikhe , completed his Masters & PhD in Prosthodontics from Rural Dental College, Loni securing third rank in the Pravara Institute of Medical Sciences Deemed University. He was awarded Dr.G.C.DAS Memorial Award for Research on Implants at 39th IPS conference Dubai (U A E).He has two patents under his name. He has received Dr.Saraswati medal award for best research for implant study in 2017.He has received Fully funded scholarship to Spain ,university of Santiago de Compostela. He has completed fellowship in Implantlogy from Noble Biocare. \nHe has attended various conferences and CDE programmes and has national publications to his credit. His field of interest is in Implant supported prosthesis. Presently he is working as a associate professor in the Dept of Prosthodontics, Rural Dental College, Loni and maintains a successful private practice specialising in Implantology at Rahata.\n\nEmail: drdeepak_mvikhe@yahoo.com..................",institutionString:null,institution:{name:"Pravara Institute of Medical Sciences",country:{name:"India"}}},{id:"204110",title:"Dr.",name:"Ahmed A.",middleName:null,surname:"Madfa",slug:"ahmed-a.-madfa",fullName:"Ahmed A. Madfa",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/204110/images/system/204110.jpg",biography:"Dr. Madfa is currently Associate Professor of Endodontics at Thamar University and a visiting lecturer at Sana'a University and University of Sciences and Technology. He has more than 6 years of experience in teaching. His research interests include root canal morphology, functionally graded concept, dental biomaterials, epidemiology and dental education, biomimetic restoration, finite element analysis and endodontic regeneration. Dr. Madfa has numerous international publications, full articles, two patents, a book and a book chapter. Furthermore, he won 14 international scientific awards. Furthermore, he is involved in many academic activities ranging from editorial board member, reviewer for many international journals and postgraduate students' supervisor. Besides, I deliver many courses and training workshops at various scientific events. Dr. Madfa also regularly attends international conferences and holds administrative positions (Deputy Dean of the Faculty for Students’ & Academic Affairs and Deputy Head of Research Unit).",institutionString:"Thamar University",institution:null},{id:"210472",title:"Dr.",name:"Nermin",middleName:"Mohammed Ahmed",surname:"Yussif",slug:"nermin-yussif",fullName:"Nermin Yussif",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/210472/images/system/210472.jpg",biography:"Dr. Nermin Mohammed Ahmed Yussif is working at the Faculty of dentistry, University for October university for modern sciences and arts (MSA). Her areas of expertise include: periodontology, dental laserology, oral implantology, periodontal plastic surgeries, oral mesotherapy, nutrition, dental pharmacology. She is an editor and reviewer in numerous international journals.",institutionString:"MSA University",institution:null},{id:"204606",title:"Dr.",name:"Serdar",middleName:null,surname:"Gözler",slug:"serdar-gozler",fullName:"Serdar Gözler",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/204606/images/system/204606.jpeg",biography:"Dr. Serdar Gözler has completed his undergraduate studies at the Marmara University Faculty of Dentistry in 1978, followed by an assistantship in the Prosthesis Department of Dicle University Faculty of Dentistry. Starting his PhD work on non-resilient overdentures with Assoc. Prof. Hüsnü Yavuzyılmaz, he continued his studies with Prof. Dr. Gürbüz Öztürk of Istanbul University Faculty of Dentistry Department of Prosthodontics, this time on Gnatology. He attended training programs on occlusion, neurology, neurophysiology, EMG, radiology and biostatistics. In 1982, he presented his PhD thesis \\Gerber and Lauritzen Occlusion Analysis Techniques: Diagnosis Values,\\ at Istanbul University School of Dentistry, Department of Prosthodontics. As he was also working with Prof. Senih Çalıkkocaoğlu on The Physiology of Chewing at the same time, Gözler has written a chapter in Çalıkkocaoğlu\\'s book \\Complete Prostheses\\ entitled \\The Place of Neuromuscular Mechanism in Prosthetic Dentistry.\\ The book was published five times since by the Istanbul University Publications. Having presented in various conferences about occlusion analysis until 1998, Dr. Gözler has also decided to use the T-Scan II occlusion analysis method. Having been personally trained by Dr. Robert Kerstein on this method, Dr. Gözler has been lecturing on the T-Scan Occlusion Analysis Method in conferences both in Turkey and abroad. Dr. Gözler has various articles and presentations on Digital Occlusion Analysis methods. He is now Head of the TMD Clinic at Prosthodontic Department of Faculty of Dentistry , Istanbul Aydın University , Turkey.",institutionString:"Istanbul Aydin University",institution:{name:"Istanbul Aydın University",country:{name:"Turkey"}}},{id:"240870",title:"Ph.D.",name:"Alaa Eddin Omar",middleName:null,surname:"Al Ostwani",slug:"alaa-eddin-omar-al-ostwani",fullName:"Alaa Eddin Omar Al Ostwani",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/240870/images/system/240870.jpeg",biography:"Dr. Al Ostwani Alaa Eddin Omar received his Master in dentistry from Damascus University in 2010, and his Ph.D. in Pediatric Dentistry from Damascus University in 2014. Dr. Al Ostwani is an assistant professor and faculty member at IUST University since 2014. \nDuring his academic experience, he has received several awards including the scientific research award from the Union of Arab Universities, the Syrian gold medal and the international gold medal for invention and creativity. Dr. Al Ostwani is a Member of the International Association of Dental Traumatology and the Syrian Society for Research and Preventive Dentistry since 2017. He is also a Member of the Reviewer Board of International Journal of Dental Medicine (IJDM), and the Indian Journal of Conservative and Endodontics since 2016.",institutionString:"International University for Science and Technology.",institution:{name:"Islamic University of Science and Technology",country:{name:"India"}}},{id:"42847",title:"Dr.",name:"Belma",middleName:null,surname:"Işik Aslan",slug:"belma-isik-aslan",fullName:"Belma Işik Aslan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/42847/images/system/42847.jpg",biography:"Dr. Belma IşIk Aslan was born in 1976 in Ankara-TURKEY. After graduating from TED Ankara College in 1994, she attended to Gazi University, Faculty of Dentistry in Ankara. She completed her PhD in orthodontic education at Gazi University between 1999-2005. Dr. Işık Aslan stayed at the Providence Hospital Craniofacial Institude and Reconstructive Surgery in Michigan, USA for three months as an observer. She worked as a specialist doctor at Gazi University, Dentistry Faculty, Department of Orthodontics between 2005-2014. She was appointed as associate professor in January, 2014 and as professor in 2021. Dr. Işık Aslan still works as an instructor at the same faculty. She has published a total of 35 articles, 10 book chapters, 39 conference proceedings both internationally and nationally. Also she was the academic editor of the international book 'Current Advances in Orthodontics'. She is a member of the Turkish Orthodontic Society and Turkish Cleft Lip and Palate Society. She is married and has 2 children. Her knowledge of English is at an advanced level.",institutionString:"Gazi University Dentistry Faculty Department of Orthodontics",institution:null},{id:"178412",title:"Associate Prof.",name:"Guhan",middleName:null,surname:"Dergin",slug:"guhan-dergin",fullName:"Guhan Dergin",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/178412/images/6954_n.jpg",biography:"Assoc. Prof. Dr. Gühan Dergin was born in 1973 in Izmit. He graduated from Marmara University Faculty of Dentistry in 1999. He completed his specialty of OMFS surgery in Marmara University Faculty of Dentistry and obtained his PhD degree in 2006. In 2005, he was invited as a visiting doctor in the Oral and Maxillofacial Surgery Department of the University of North Carolina, USA, where he went on a scholarship. Dr. Dergin still continues his academic career as an associate professor in Marmara University Faculty of Dentistry. He has many articles in international and national scientific journals and chapters in books.",institutionString:null,institution:{name:"Marmara University",country:{name:"Turkey"}}},{id:"178414",title:"Prof.",name:"Yusuf",middleName:null,surname:"Emes",slug:"yusuf-emes",fullName:"Yusuf Emes",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/178414/images/6953_n.jpg",biography:"Born in Istanbul in 1974, Dr. Emes graduated from Istanbul University Faculty of Dentistry in 1997 and completed his PhD degree in Istanbul University faculty of Dentistry Department of Oral and Maxillofacial Surgery in 2005. He has papers published in international and national scientific journals, including research articles on implantology, oroantral fistulas, odontogenic cysts, and temporomandibular disorders. Dr. Emes is currently working as a full-time academic staff in Istanbul University faculty of Dentistry Department of Oral and Maxillofacial Surgery.",institutionString:null,institution:{name:"Istanbul University",country:{name:"Turkey"}}},{id:"192229",title:"Ph.D.",name:"Ana Luiza",middleName:null,surname:"De Carvalho Felippini",slug:"ana-luiza-de-carvalho-felippini",fullName:"Ana Luiza De Carvalho Felippini",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/192229/images/system/192229.jpg",biography:null,institutionString:"University of São Paulo",institution:{name:"University of Sao Paulo",country:{name:"Brazil"}}},{id:"256851",title:"Prof.",name:"Ayşe",middleName:null,surname:"Gülşen",slug:"ayse-gulsen",fullName:"Ayşe Gülşen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/256851/images/9696_n.jpg",biography:"Dr. Ayşe Gülşen graduated in 1990 from Faculty of Dentistry, University of Ankara and did a postgraduate program at University of Gazi. \nShe worked as an observer and research assistant in Craniofacial Surgery Departments in New York, Providence Hospital in Michigan and Chang Gung Memorial Hospital in Taiwan. \nShe works as Craniofacial Orthodontist in Department of Aesthetic, Plastic and Reconstructive Surgery, Faculty of Medicine, University of Gazi, Ankara Turkey since 2004.",institutionString:"Univeristy of Gazi",institution:null},{id:"255366",title:"Prof.",name:"Tosun",middleName:null,surname:"Tosun",slug:"tosun-tosun",fullName:"Tosun Tosun",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/255366/images/7347_n.jpg",biography:"Graduated at the Faculty of Dentistry, University of Istanbul, Turkey in 1989;\nVisitor Assistant at the University of Padua, Italy and Branemark Osseointegration Center of Treviso, Italy between 1993-94;\nPhD thesis on oral implantology in University of Istanbul and was awarded the academic title “Dr.med.dent.”, 1997;\nHe was awarded the academic title “Doç.Dr.” (Associated Professor) in 2003;\nProficiency in Botulinum Toxin Applications, Reading-UK in 2009;\nMastership, RWTH Certificate in Laser Therapy in Dentistry, AALZ-Aachen University, Germany 2009-11;\nMaster of Science (MSc) in Laser Dentistry, University of Genoa, Italy 2013-14.\n\nDr.Tosun worked as Research Assistant in the Department of Oral Implantology, Faculty of Dentistry, University of Istanbul between 1990-2002. \nHe worked part-time as Consultant surgeon in Harvard Medical International Hospitals and John Hopkins Medicine, Istanbul between years 2007-09.\u2028He was contract Professor in the Department of Surgical and Diagnostic Sciences (DI.S.C.), Medical School, University of Genova, Italy between years 2011-16. \nSince 2015 he is visiting Professor at Medical School, University of Plovdiv, Bulgaria. \nCurrently he is Associated Prof.Dr. at the Dental School, Oral Surgery Dept., Istanbul Aydin University and since 2003 he works in his own private clinic in Istanbul, Turkey.\u2028\nDr.Tosun is reviewer in journal ‘Laser in Medical Sciences’, reviewer in journal ‘Folia Medica\\', a Fellow of the International Team for Implantology, Clinical Lecturer of DGZI German Association of Oral Implantology, Expert Lecturer of Laser&Health Academy, Country Representative of World Federation for Laser Dentistry, member of European Federation of Periodontology, member of Academy of Laser Dentistry. Dr.Tosun presents papers in international and national congresses and has scientific publications in international and national journals. He speaks english, spanish, italian and french.",institutionString:null,institution:{name:"Istanbul Aydın University",country:{name:"Turkey"}}},{id:"171887",title:"Prof.",name:"Zühre",middleName:null,surname:"Akarslan",slug:"zuhre-akarslan",fullName:"Zühre Akarslan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/171887/images/system/171887.jpg",biography:"Zühre Akarslan was born in 1977 in Cyprus. She graduated from Gazi University Faculty of Dentistry, Ankara, Turkey in 2000. \r\nLater she received her Ph.D. degree from the Oral Diagnosis and Radiology Department; which was recently renamed as Oral and Dentomaxillofacial Radiology, from the same university. \r\nShe is working as a full-time Associate Professor and is a lecturer and an academic researcher. \r\nHer expertise areas are dental caries, cancer, dental fear and anxiety, gag reflex in dentistry, oral medicine, and dentomaxillofacial radiology.",institutionString:"Gazi University",institution:{name:"Gazi University",country:{name:"Turkey"}}},{id:"256417",title:"Associate Prof.",name:"Sanaz",middleName:null,surname:"Sadry",slug:"sanaz-sadry",fullName:"Sanaz Sadry",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/256417/images/8106_n.jpg",biography:null,institutionString:null,institution:null},{id:"272237",title:"Dr.",name:"Pinar",middleName:"Kiymet",surname:"Karataban",slug:"pinar-karataban",fullName:"Pinar Karataban",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/272237/images/8911_n.png",biography:"Assist.Prof.Dr.Pınar Kıymet Karataban, DDS PhD \n\nDr.Pınar Kıymet Karataban was born in Istanbul in 1975. After her graduation from Marmara University Faculty of Dentistry in 1998 she started her PhD in Paediatric Dentistry focused on children with special needs; mainly children with Cerebral Palsy. She finished her pHD thesis entitled \\'Investigation of occlusion via cast analysis and evaluation of dental caries prevalance, periodontal status and muscle dysfunctions in children with cerebral palsy” in 2008. She got her Assist. Proffessor degree in Istanbul Aydın University Paediatric Dentistry Department in 2015-2018. ın 2019 she started her new career in Bahcesehir University, Istanbul as Head of Department of Pediatric Dentistry. In 2020 she was accepted to BAU International University, Batumi as Professor of Pediatric Dentistry. She’s a lecturer in the same university meanwhile working part-time in private practice in Ege Dental Studio (https://www.egedisklinigi.com/) a multidisciplinary dental clinic in Istanbul. Her main interests are paleodontology, ancient and contemporary dentistry, oral microbiology, cerebral palsy and special care dentistry. She has national and international publications, scientific reports and is a member of IAPO (International Association for Paleodontology), IADH (International Association of Disability and Oral Health) and EAPD (European Association of Pediatric Dentistry).",institutionString:null,institution:null},{id:"202198",title:"Dr.",name:"Buket",middleName:null,surname:"Aybar",slug:"buket-aybar",fullName:"Buket Aybar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/202198/images/6955_n.jpg",biography:"Buket Aybar, DDS, PhD, was born in 1971. She graduated from Istanbul University, Faculty of Dentistry, in 1992 and completed her PhD degree on Oral and Maxillofacial Surgery in Istanbul University in 1997.\nDr. Aybar is currently a full-time professor in Istanbul University, Faculty of Dentistry Department of Oral and Maxillofacial Surgery. She has teaching responsibilities in graduate and postgraduate programs. Her clinical practice includes mainly dentoalveolar surgery.\nHer topics of interest are biomaterials science and cell culture studies. She has many articles in international and national scientific journals and chapters in books; she also has participated in several scientific projects supported by Istanbul University Research fund.",institutionString:null,institution:null},{id:"260116",title:"Dr.",name:"Mehmet",middleName:null,surname:"Yaltirik",slug:"mehmet-yaltirik",fullName:"Mehmet Yaltirik",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/260116/images/7413_n.jpg",biography:"Birth Date 25.09.1965\r\nBirth Place Adana- Turkey\r\nSex Male\r\nMarrial Status Bachelor\r\nDriving License Acquired\r\nMother Tongue Turkish\r\n\r\nAddress:\r\nWork:University of Istanbul,Faculty of Dentistry, Department of Oral Surgery and Oral Medicine 34093 Capa,Istanbul- TURKIYE",institutionString:null,institution:null},{id:"172009",title:"Dr.",name:"Fatma Deniz",middleName:null,surname:"Uzuner",slug:"fatma-deniz-uzuner",fullName:"Fatma Deniz Uzuner",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/172009/images/7122_n.jpg",biography:"Dr. Deniz Uzuner was born in 1969 in Kocaeli-TURKEY. After graduating from TED Ankara College in 1986, she attended the Hacettepe University, Faculty of Dentistry in Ankara. \nIn 1993 she attended the Gazi University, Faculty of Dentistry, Department of Orthodontics for her PhD education. After finishing the PhD education, she worked as orthodontist in Ankara Dental Hospital under the Turkish Government, Ministry of Health and in a special Orthodontic Clinic till 2011. Between 2011 and 2016, Dr. Deniz Uzuner worked as a specialist in the Department of Orthodontics, Faculty of Dentistry, Gazi University in Ankara/Turkey. In 2016, she was appointed associate professor. Dr. Deniz Uzuner has authored 23 Journal Papers, 3 Book Chapters and has had 39 oral/poster presentations. She is a member of the Turkish Orthodontic Society. Her knowledge of English is at an advanced level.",institutionString:null,institution:null},{id:"332914",title:"Dr.",name:"Muhammad Saad",middleName:null,surname:"Shaikh",slug:"muhammad-saad-shaikh",fullName:"Muhammad Saad Shaikh",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Jinnah Sindh Medical University",country:{name:"Pakistan"}}},{id:"315775",title:"Dr.",name:"Feng",middleName:null,surname:"Luo",slug:"feng-luo",fullName:"Feng Luo",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Sichuan University",country:{name:"China"}}},{id:"423519",title:"Dr.",name:"Sizakele",middleName:null,surname:"Ngwenya",slug:"sizakele-ngwenya",fullName:"Sizakele Ngwenya",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of the Witwatersrand",country:{name:"South Africa"}}},{id:"419270",title:"Dr.",name:"Ann",middleName:null,surname:"Chianchitlert",slug:"ann-chianchitlert",fullName:"Ann Chianchitlert",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Walailak University",country:{name:"Thailand"}}},{id:"419271",title:"Dr.",name:"Diane",middleName:null,surname:"Selvido",slug:"diane-selvido",fullName:"Diane Selvido",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Walailak University",country:{name:"Thailand"}}},{id:"419272",title:"Dr.",name:"Irin",middleName:null,surname:"Sirisoontorn",slug:"irin-sirisoontorn",fullName:"Irin Sirisoontorn",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Walailak University",country:{name:"Thailand"}}},{id:"355660",title:"Dr.",name:"Anitha",middleName:null,surname:"Mani",slug:"anitha-mani",fullName:"Anitha Mani",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"SRM Dental College",country:{name:"India"}}},{id:"355612",title:"Dr.",name:"Janani",middleName:null,surname:"Karthikeyan",slug:"janani-karthikeyan",fullName:"Janani Karthikeyan",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"SRM Dental College",country:{name:"India"}}},{id:"334400",title:"Dr.",name:"Suvetha",middleName:null,surname:"Siva",slug:"suvetha-siva",fullName:"Suvetha Siva",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"SRM Dental College",country:{name:"India"}}},{id:"334239",title:"Prof.",name:"Leung",middleName:null,surname:"Wai Keung",slug:"leung-wai-keung",fullName:"Leung Wai Keung",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Hong Kong",country:{name:"China"}}}]}},subseries:{item:{id:"4",type:"subseries",title:"Fungal Infectious Diseases",keywords:"Emerging Fungal Pathogens, Invasive Infections, Epidemiology, Cell Membrane, Fungal Virulence, Diagnosis, Treatment",scope:"Fungi are ubiquitous and there are almost no non-pathogenic fungi. Fungal infectious illness prevalence and prognosis are determined by the exposure between fungi and host, host immunological state, fungal virulence, and early and accurate diagnosis and treatment. \r\nPatients with both congenital and acquired immunodeficiency are more likely to be infected with opportunistic mycosis. Fungal infectious disease outbreaks are common during the post- disaster rebuilding era, which is characterised by high population density, migration, and poor health and medical conditions.\r\nSystemic or local fungal infection is mainly associated with the fungi directly inhaled or inoculated in the environment during the disaster. The most common fungal infection pathways are human to human (anthropophilic), animal to human (zoophilic), and environment to human (soilophile). Diseases are common as a result of widespread exposure to pathogenic fungus dispersed into the environment. \r\nFungi that are both common and emerging are intertwined. In Southeast Asia, for example, Talaromyces marneffei is an important pathogenic thermally dimorphic fungus that causes systemic mycosis. Widespread fungal infections with complicated and variable clinical manifestations, such as Candida auris infection resistant to several antifungal medicines, Covid-19 associated with Trichoderma, and terbinafine resistant dermatophytosis in India, are among the most serious disorders. \r\nInappropriate local or systemic use of glucocorticoids, as well as their immunosuppressive effects, may lead to changes in fungal infection spectrum and clinical characteristics. Hematogenous candidiasis is a worrisome issue that affects people all over the world, particularly ICU patients. CARD9 deficiency and fungal infection have been major issues in recent years. Invasive aspergillosis is associated with a significant death rate. Special attention should be given to endemic fungal infections, identification of important clinical fungal infections advanced in yeasts, filamentous fungal infections, skin mycobiome and fungal genomes, and immunity to fungal infections.\r\nIn addition, endemic fungal diseases or uncommon fungal infections caused by Mucor irregularis, dermatophytosis, Malassezia, cryptococcosis, chromoblastomycosis, coccidiosis, blastomycosis, histoplasmosis, sporotrichosis, and other fungi, should be monitored. \r\nThis topic includes the research progress on the etiology and pathogenesis of fungal infections, new methods of isolation and identification, rapid detection, drug sensitivity testing, new antifungal drugs, schemes and case series reports. It will provide significant opportunities and support for scientists, clinical doctors, mycologists, antifungal drug researchers, public health practitioners, and epidemiologists from all over the world to share new research, ideas and solutions to promote the development and progress of medical mycology.",coverUrl:"https://cdn.intechopen.com/series_topics/covers/4.jpg",hasOnlineFirst:!0,hasPublishedBooks:!1,annualVolume:11400,editor:{id:"174134",title:"Dr.",name:"Yuping",middleName:null,surname:"Ran",slug:"yuping-ran",fullName:"Yuping Ran",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bS9d6QAC/Profile_Picture_1630330675373",biography:"Dr. Yuping Ran, Professor, Department of Dermatology, West China Hospital, Sichuan University, Chengdu, China. Completed the Course Medical Mycology, the Centraalbureau voor Schimmelcultures (CBS), Fungal Biodiversity Centre, Netherlands (2006). International Union of Microbiological Societies (IUMS) Fellow, and International Emerging Infectious Diseases (IEID) Fellow, Centers for Diseases Control and Prevention (CDC), Atlanta, USA. Diploma of Dermatological Scientist, Japanese Society for Investigative Dermatology. Ph.D. of Juntendo University, Japan. Bachelor’s and Master’s degree, Medicine, West China University of Medical Sciences. Chair of Sichuan Medical Association Dermatology Committee. General Secretary of The 19th Annual Meeting of Chinese Society of Dermatology and the Asia Pacific Society for Medical Mycology (2013). In charge of the Annual Medical Mycology Course over 20-years authorized by National Continue Medical Education Committee of China. Member of the board of directors of the Asia-Pacific Society for Medical Mycology (APSMM). Associate editor of Mycopathologia. Vice-chief of the editorial board of Chinses Journal of Mycology, China. Board Member and Chair of Mycology Group of Chinese Society of Dermatology.",institutionString:null,institution:{name:"Sichuan University",institutionURL:null,country:{name:"China"}}},editorTwo:null,editorThree:null,series:{id:"6",title:"Infectious Diseases",doi:"10.5772/intechopen.71852",issn:"2631-6188"},editorialBoard:[{id:"302145",title:"Dr.",name:"Felix",middleName:null,surname:"Bongomin",slug:"felix-bongomin",fullName:"Felix Bongomin",profilePictureURL:"https://mts.intechopen.com/storage/users/302145/images/system/302145.jpg",institutionString:null,institution:{name:"Gulu University",institutionURL:null,country:{name:"Uganda"}}},{id:"45803",title:"Ph.D.",name:"Payam",middleName:null,surname:"Behzadi",slug:"payam-behzadi",fullName:"Payam Behzadi",profilePictureURL:"https://mts.intechopen.com/storage/users/45803/images/system/45803.jpg",institutionString:"Islamic Azad University, Tehran",institution:{name:"Islamic Azad University, Tehran",institutionURL:null,country:{name:"Iran"}}}]},onlineFirstChapters:{paginationCount:14,paginationItems:[{id:"82103",title:"The Role of Endoplasmic Reticulum Stress and Its Regulation in the Progression of Neurological and Infectious Diseases",doi:"10.5772/intechopen.105543",signatures:"Mary Dover, Michael Kishek, Miranda Eddins, Naneeta Desar, Ketema Paul and Milan Fiala",slug:"the-role-of-endoplasmic-reticulum-stress-and-its-regulation-in-the-progression-of-neurological-and-i",totalDownloads:5,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Updates on Endoplasmic Reticulum",coverURL:"https://cdn.intechopen.com/books/images_new/11674.jpg",subseries:{id:"14",title:"Cell and Molecular Biology"}}},{id:"80954",title:"Ion Channels and Neurodegenerative Disease Aging Related",doi:"10.5772/intechopen.103074",signatures:"Marika Cordaro, Salvatore Cuzzocrea and Rosanna Di Paola",slug:"ion-channels-and-neurodegenerative-disease-aging-related",totalDownloads:6,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Ion Channels - From Basic Properties to Medical Treatment",coverURL:"https://cdn.intechopen.com/books/images_new/10838.jpg",subseries:{id:"14",title:"Cell and Molecular Biology"}}},{id:"81647",title:"Diabetes and Epigenetics",doi:"10.5772/intechopen.104653",signatures:"Rasha A. 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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. 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