Monoclonal antibody-based therapeutic drugs approved by FDA (Food and Drug Administration) until 2015.
\r\n\t
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In recent years, the use of mAbs has been expanded due to significant advances in design. The effect of decreasing immunogenicity in humans, improvement in their bioavailability, optimizing the affinity and antigen-binding specificity, and other advances in protein engineering are improving therapeutic mAb profiles (Figure 1) [2].
\nSchematic overview of a monoclonal antibody, showing their heavy and variable chain.
With the advent of genetic engineering, it has been possible to develop new methods to obtain monoclonal antibodies, both for improvement with regard to these humanized antibodies and for production models [4–6]. Advances in molecular and cell biology for the development of more efficient antibodies have allowed advances in diagnostic and therapeutic areas. Such advances have triggered improvements in production processes, allowing for the reduction of production costs and thus leading to an increase in the popularization of treatments with mAbs. All process improvements provide a consistent and reproducible production of large quantities of mAbs at a moderate cost [4–6].
\nLarge-scale production has revolutionized the market for monoclonal antibodies by boosting its production, making this a more practical method of production. Production techniques have only had a sizable breakthrough due to molecular techniques [1, 7].
\nIn general, a process of commercial production of mAb begins with the generation of an mAb by immunizing an animal or by molecular biology methods involving the identification and optimization of the coding DNA sequence and the construction and identification of a stable high-producing clone. Improvements in cultivation are similar to those applied in other bioproducts that rely on culturing microorganisms or cells, requiring the development of a well-designed culturing process comprising the full range of control and associated operations that will support technical evaluations [1, 8].
\nmAbs production processes in wave or single-use bioreactor (SUBs) are characterized by flexibility and low operating costs when compared to the production processes in fixed stainless steel vats. The development of bioprocesses involving these production platforms can reap greater acceptance by the industry [9–11].
\nDrugs based on mAbs have been controlled by regulatory agencies around the world. Therefore, it is necessary to elaborate regulatory protocols accompanying the increase in production and the nuances of the characteristics of this class of drugs [10, 11].
\nThe proposed chapter covers the fundamental aspects of monoclonal antibody production methods, with emphasis on methodologies using immobilized cells, wave bioreactor systems, SUBs, and finally the roller bottles technique. Such techniques have been described in the most recent literature, both for murine monoclonal antibody production and for production of antibodies from modified microorganisms.
\nMilstein and Köhler described the first technique developed for stable monoclonal antibody production in 1975. This technique consists of creating a hybridoma, a stable hybrid cell capable of producing a single type of antibody against a specific epitope present in an antigen. Hybridoma construction was initially produced from murine models. The technique consists of removing a pool of activated B lymphocytes from an immunized animal spleen and combining them with immortalized myeloma cells unable to produce the enzyme hypoxanthine-guanine-phosphoribosyltransferase (HGPRT), an important enzyme present in the
In spite of the fact that the primary recombinant mAbs were delivered utilizing this innovation—including the first medication approved by the Food and Drug Administration (FDA) for therapeutic proposes (Table 1)—the great contribution of this technology was mostly to elucidate immune response mechanisms and control in vitro antibody production. Therefore, mAb hybridoma production from murine sources exhibits a genuine downside in human therapeutics (Figure 1).
\nDrug name | \nActive ingredient | \nDescription | \nTarget | \nTherapeutic category | \napproval (FDA) | \n
---|---|---|---|---|---|
ACTEMRA® | \nTocilizumab | \nHumanized IgG1κ | \nIL-6 receptor | \nImmunological | \n2010 | \n
ADCETRIS® | \nBrentuximab vedotin | \nChimeric IgG1 | \nCD30 | \nCancer | \n2011 | \n
ARZERRA® | \nOfatumumab | \nHuman IgG1κ | \nCD20 | \nCancer | \n2009 | \n
AVASTIN® | \nBevacizumab | \nHumanized IgG1 | \nVEGF | \nCancer | \n2004 | \n
BENLYSTA® | \nBelimumab | \nHuman IgG1λ | \nBLyS | \nImmunological | \n2011 | \n
BEXXAR | \nTositumomab; iodine I 131 tositumomab | \nIgG2αλ, I131 | \nCD20 | \nCancer | \n2003 | \n
BLINCYTO | \nBlinatumomab | \nBiTE antibody-scFvs | \nCD19/CD3 | \nCancer | \n2014 | \n
CAMPATH (LEMTRADA™) | \nAlemtuzumab | \nHumanized IgG1κ | \nCD52 | \nImmunological | \n2001 | \n
CEA-SCAN | \nArcitumomab | \nMurine IgG1 Fab’ | \nCEA | \nDiagnosys | \n1996 | \n
CIMZIA® | \nCertolizumab pegol | \nHumanized Fab’, PEG | \nTNFα | \nImmunological | \n2008 | \n
COSENTYX® | \nSecukinumab | \nHuman IgG1κ | \nIL-17A | \nImmunological | \n2015 | \n
CYRAMZA | \nRamucirumab | \nHuman IgG1 | \nVEGRF-2 | \nCancer | \n2014 | \n
DARZALEX | \nDaratumumab | \nHuman IgG1κ | \nCD38 | \nCancer | \n2015 | \n
HERCEPTIN® | \nTrastuzumab | \nhumanized IgG1κ | \nHER2 | \nCancer | \n1998 | \n
EMPLICITI™ | \nElotuzumab | \nHumanized IgG1 | \nSLAMF7 | \nCancer | \n2015 | \n
ENTYVIO | \nVedolizumab | \nHumanized IgG1 | \nα4β7 Integrin | \nImmunological | \n2014 | \n
ERBITUX® | \nCetuximab | \nChimeric IgG1 | \nEGFR | \nCancer | \n2004 | \n
GAZYVA® | \nObinutuzumab | \nHumanized IgG1 | \nCD20 | \nCancer | \n2013 | \n
HUMIRA | \nAdalimumab | \nHuman IgG1 | \nTNF | \nImmunological | \n2002 | \n
ILARIS | \nCanakinumab | \nHuman IgG1κ | \numan-IL-1β | \nImmunological/anti-inflammatory | \n2009 | \n
KADCYLA® | \nAdo-trastuzumab emtansine | \nHumanized IgG1; DM1 | \nHER2 | \nCancer | \n2013 | \n
KEYTRUDA® | \nPembrolizumab | \nHumanized IgG4κ | \nPD-1 | \nCancer | \n2014 | \n
LEMTRADA™ | \nAlemtuzumab | \nHumanized IgG1κ | \nCD52 | \nImmunological | \n2001 | \n
LUCENTIS | \nRanibizumab | \nHumanized IgG1κ | \nVEGF-A | \nOphthalmic | \n2006 | \n
Muromomab | \nOrthoclone | \nMurine IgG2α | \nCD3 | \nImmunological | \n1992 | \n
Mylotarg® | \nGemtuzumab ozogamicin | \nHumanized IgG4κ, calicheamicin | \nCD33 | \nCancer | \n2000 | \n
MYOSCINT® | \nImciromab Penlelale | \nMurine IgG2/4κ Fab’;DTPA | \nHeavy chain of human myosin | \nDetection of myocardial injury | \n1996 | \n
NUCALA® | \nMepolizumab | \nHumanized IgG1κ | \nIL-5 | \nImmunological | \n2015 | \n
OPDIVO | \nNivolumab | \nHuman IgG4κ | \nPD-1 | \nCancer | \n2014 | \n
PERJETA® | \nPertuzumab | \nHumanized IgG | \nHER2/neu receptor | \nCancer | \n2012 | \n
PORTRAZZA | \nNecitumumab | \nHuman IgGκ | \nEGFR | \nCancer | \n2015 | \n
PRALUENT™ | \nAlirocumab | \nHuman IgG1 | \nPCSK9 | \nLipid-lowering | \n2015 | \n
PRAXBIND® | \nIdarucizumab | \nHumanized IgG1 Fab | \nDabigatran (anticoagulant) | \nHemostasis | \n2015 | \n
XGEVA® | \nDenosumab | \nHuman IgG2 | \nRANKL | \nBone disorders | \n2010 | \n
ProstaScint® | \nCapromab pendetide | \nMurine IgG1κ, GYK-DTPA-HCl | \nPSMA | \nCancer | \n1996 | \n
RAPTIVA® | \nEfalizumab | \nHumanized IgG1κ | \nCD11a | \nImmunological | \n2003 | \n
RAXIBACUMAB | \nRaxibacumab | \nHuman IgG1λ | \nPA of B. Anthracis toxin | \nAnti-toxin | \n2012 | \n
REMICADE® | \nInfliximab | \nChimeric IgG1κ | \nTNFα | \nImmunological | \n1998 | \n
ReoPro® | \nAbciximab | \nChimeric IgG1κ Fab | \nGPIIb/IIIa | \nHemostasis | \n1993 | \n
REPATHA | \nEvolocumab | \nHuman IgG2 | \nPCSK9 | \nLipid-lowering | \n2015 | \n
RITUXAN® | \nRituximab | \nChimeric IgG1κ | \nCD20 | \nCancer | \n1997 | \n
SIMPONI | \nGolimumab | \nHuman IgG1κ | \nTNFα | \nImmunological | \n2009 | \n
SIMULECT® | \nBasiliximab | \nChimeric IgG1κ | \nIL-2 receptor | \nImmunological | \n1998 | \n
SOLIRIS® | \nEculizumab | \nHumanized IgG2/4κ | \nC5 | \nHemostasis | \n2007 | \n
STELARA® | \nUstekinumab | \nHuman IgG1κ | \nIL-12 and IL 23 | \nImmunological | \n2009 | \n
SYLVANT | \nSiltuximab | \nChimeric IgG | \nIL-6 | \nImmunological | \n2014 | \n
SYNAGIS® | \nPalivizumab | \nHumanized IgG1κ | \nRSV F | \nAntiviral | \n1998 | \n
NeutroSpec™ | \nFanolesomaB; technetium Tc 99m | \nMurine IgM | \n3-fucosyl-N-acetyllactosamine | \nDiagnosys | \n2004 | \n
TYSABRI | \nNatalizumab | \nHumanized IgG4κ | \nα4β1/α4β7 integrins | \nImmunological | \n2004 | \n
UNITUXIN™ | \nDinutuximab | \nChimeric IgG1κ | \nGlycolipid GD2 | \nCancer | \n2015 | \n
VECTIBIX® | \nPanitumumab | \nHuman IgG2κ | \nEGFR | \nCancer | \n2006 | \n
VERLUMA™ | \nNofetumomab | \nMurine IgG2b Fab | \nGlycoprotein antigen expressed in a variety of cancers | \nDiagnosys | \n1996 | \n
XGEVA | \nDenosumab | \nHuman IgG2 | \nRANKL | \nCancer | \n2010 | \n
XOLAIR® | \nOmalizumab | \nHumanized IgG1κ | \nHuman IgE | \nImmunological | \n2003 | \n
YERVOY® | \nIpilimumab | \nHuman IgG1κ | \nCTLA-4 | \nCancer | \n2011 | \n
ZENAPAX® | \nDaclizumab | \nHumanized IgG1 | \nIL-2 receptor | \nImmunological | \n1997 | \n
ZEVALIN® | \nIbritumomab tiuxetan | \nmurine IgG1κ, Yttrium-90 | \nCD20 | \nCancer | \n2002 | \n
Monoclonal antibody-based therapeutic drugs approved by FDA (Food and Drug Administration) until 2015.
After a few infusions, murine antibody molecules trigger the human anti-mouse antibody (HAMA) response of the human immune system [1, 12]. To work around this issue, new methodologies have been developed to deliver antibodies similar to human molecules, so the technology evolved to less immunogenic chimeric antibodies (constant regions of human antibodies linked to the variable region of the murine source), creating a new set of therapeutic possibilities (Figure 1). Subsequently, the need for an even less immunogenic alternative boosted the production of humanized antibodies (only the region that interacts with the antigen epitope is from mouse origin) (Figure 1). Even fully human antibodies (Figure 1) can be produced from genetically modified mice [13].
\nA great improvement in mAb production has come with the development of phage display libraries. This methodology helps to investigate interactions between molecules (protein-protein, protein-peptide, and protein-DNA) and consists, basically, in cloning Fab-region-coding genes amplified from B lymphocytes into bacteriophage plasmid vectors. Then the bacterium can be transformed with these vectors, going on to express the heterologous genes from a viral capsid. This capsid contains viral proteins and proteins encoded by the Fab sequence received by that specific cell. Once the library is complete, the affinity between proteins produced from different Fab regions can be tested against the antigen of interest and the cell transformed with the plasmid that contains those genes can be readily sequenced. The advantages of this methodology are the following: the same library has the potential to generate a great number of new antibodies, it is an in vitro process, so it does not require animal immunizations steps, and because of that, toxic antigens can be tested. Also, a greater variety of antigens can be tested, and antibody molecules can be rapidly obtained [13].
\nOne of the most critical steps in developing an mAb production system is to choose the cell line. The cells must be stable and secrete the desired protein with the correct conformation at high levels. Based on these requirements, the mammalian cell is the most commonly chosen expression system for mAb production. The main advantage of a mammalian expression system is that the cellular machinery is adapted for the production, processing, and secretion of highly complex molecules. The great majority of commercial mAbs are produced in Chinese hamster ovary (CHO) and NS0 cells, originating from plasmacytoma cells that were modified until IgG generation in nonsecreting B cells. Genetic modifications in CHO cells have generated cell lines capable of producing a high quantity of humanized mAbs. These cell lines were able to secrete up to 100 pg/cell/day [14]. Other modifications led to a high production of a chimeric mAb, ranging from 80 to 110 pg/cell/day [15]. NS0 modifications also have been made, leading to higher mAb production rates, ranging from 20 to 50 pg/cell/day [16]. In smaller quantities, hybridoma cell lines are also used in industrial mAb production. Some hybridoma strains are reported to have a production rate up to 80 pg/cell/day [16]. In spite of this, different mammalian cell lines and even more peculiar expression systems such as genetically modified plant cells, genetically modified insect cells, and genetically modified microorganism cells have also been used in mAb production and have gained space in the biopharmaceutical industry [1, 8]
\nMicroorganisms modified by genetic engineering techniques have attracted much focus in industry, because these cells are simpler to handle and to modify when compared to animal cells. Other advantages of production methods using genetically modified microorganisms are that these cells have well-defined expression systems, and the production methodology is reproducible and easy to validate. Modified yeast cells, such as
Modified plants have also gained attention since plants are easy to cultivate and propagate. Other cultivation advantages such as cheap medium, low maintenance cost, and high production yields make plant production a cheaper alternative when compared to mammalian cell cultures [17]. However, there are some limitations—different glycosylation patterns and post-translational processing can also make plant cell utilization difficult [17].
\nCultivation media for mammalian cells must have a complex content of ingredients ranging from amino acids to trace elements. To supply the cellular demand of these nutrients, the culture medium uses serum in its composition, however, due to the emergence of diseases caused by defective prions, such as bovine spongiform encephalitis (BSE), there is a great incentive to remove any animal component of culture media composition, especially if the medium is used for industrial production of biopharmaceuticals products. This has led to the emergence of media free from any animal components, including well-defined media for CHO and NS0, the two most utilized cell types in mAb production. The development of a proper medium can be time consuming and very expensive. However, many companies prefer to develop their own production media to maintain the composition between production lots as well as develop an appropriate medium composition for the specific cell type that will be used and to achieve greater control over production. Added to this, the development of downstream processes that meet the requirement for high-purity products and tests to validate the final product quality raises the overall production cost of a drug based on monoclonal antibodies [1].
\nDespite the complexity of developing a culture medium, much progress has been made in this area, allowing for greater cell growth and increasing cell conservation time in suitable conditions for the growth and production of molecules of interest [8].
\nGrowing conditions can directly influence the cell growth and production levels of molecules of interest. Usually, mammalian cell culture conditions for mAb production are very well defined: 37 °C, pH 7.15, and dissolved O2 (OD) levels at 30–60%. CO2 level is monitored to mimic the physiological standard between 31 and 54 mmHg. However, changes in cellular conditions have shown great potential to change cellular metabolism toward cellular growth or molecule production and this can be used to increase mAb production. Bioprocesses can be designed to occur in two phases. First, cell growth is optimized to reach a certain cell density. Once this density is reached, the second phase begins and the bioreactor conditions are shifted so the cells continue to grow just at a maintenance rate and directing the metabolism toward monoclonal antibody production. Some CHO cell strains and hybridoma cells are sensitive to changes in temperature and pH. When subjected to temperature and pH values lower than those normally used, values between 30 and 35 °C and 6.7–7.0, respectively, cell growth metabolism is reduced and specific production increases. The growth metabolism reduction also contributes to lower production of some metabolic compounds which are toxic for cell cultures, allowing increased cell viability, which spend more time producing molecules of interest. A good way to monitor the growth stage of a cell culture for controlling changes in cultivation is watching the DO and pCO2 levels, which can also be adjusted to maximize the production of proteins such as mAbs [1].
\nThe cell culture for mAb production can follow three different types of processes. The simplest of them is batch production, which consists of a closed system where a bioreactor is sterilized and prepared with a medium containing all the nutrients needed for cellular growth and product manufacturing and then, cells are inoculated. There is no feeding system with fresh medium or withdrawal of spent medium. As the process runs, nutrient concentration decreases and waste metabolites are produced, lowering cell viability. In spite of being a simple process, batch is not the most suitable type of production platform for mammalian cell cultures, as the environment inside the reactor quickly becomes unfavorable for cell growth and, at the same time, waste product concentration increases. Cultivation factors such as initial nutrient concentration and waste metabolite production directly determine the maximum concentration that cells can reach in a bath culture. Generally, this type of cultivation reaches a maximum density of 1–2 × 106 cells/mL, and then the cell viability drops rapidly [1]. The production process lasts for 4–7 days, when productivity reaches certain concentration of interest [1]. Supernatant is collected and the product is recovered by downstream processes. The time that each batch takes to finish also depends on the production kinetics. If the production is growth dependent (production occurs concomitantly with cellular growth), batch processes can be stopped as soon as cells reach the stationary phase. But if the product is not associated with growth (production only starts when the growth rate decreases), the culture needs to be carried for a longer period of time since production only starts at stationary phase.
\nIn contrast to batch, a second type of production process utilized is continuous fermentation. There are two types of continuous production: chemostat cultures and perfusion cultures. Concerning chemostat cultures, fresh medium is added to the bioreactor and fermented medium is removed along with cells at a constant flow rate so that the culture volume remains unchanged. The flow rate (dilution rate) controls cellular growth and when these two variables are equal, the bioreactor reaches equilibrium—cell concentration, nutrient concentration, and product concentration are held constant. In this context, the culture can be kept in equilibrium for several months reaching a cell density of 10–30 × 106 cells/mL [1]. To avoid viable cell loss along with the constant outflow of the by-products of cell metabolism, many manufacturing plants have developed a cell-recycling system and thus, the perfusion culture method was developed where cells are kept inside the bioreactor. The disadvantages of continuous fermentation are the use of a large amount of expensive culture media and the difficulty in recovering the product, which comes out fairly diluted. These two disadvantages are consequences of the constant medium flow rate. To work around the product dilution problem, some production manufacturing plants have ultrafiltration systems which retain the product inside the bioreactor [18]. Another obstacle of this type of process is that the establishment of culture conditions for a stable industrial production plant can take months. For this to occur, the strain used must be very stable and have its physiological aspects clearly elucidated, such as growth rate, productivity, and response to certain stress conditions. It is not uncommon to hear that numerous attempts are made before the settlement of a stable production plant is achieved, but, once settled, this production process can bring many advantages, since it can be operated in smaller-volume bioreactors, and therefore have greater production flexibility.
\nThe third type of process for producing monoclonal antibodies is by far the most utilized at industrial scale, which is fed-batch process. In this process, the cell density reaches 8–12 × 106 cells/mL, and cell viability in the bioreactor is enhanced by controlled nutrient addition at specified intervals [1]. The production process can take 12–20 days [1]. Usually, the same medium used in the initial culture is also used for feeding, but in a more concentrated version. The feeding solution composition can be designed to supply the cells based on their metabolic state at different culture phases by analyzing and identifying the spent medium nutrients that are being more consumed. Furthermore, the medium used in feeding can be modified to promote cell growth or to stimulate molecule production, since different components may modify the behavior of cells, changing the metabolism for different purposes. The feed solution can also be designed to minimize the production of waste metabolites that cause cell stress when in excess. However, their production is not completely avoidable as they eventually reach harmful concentrations. It is relatively easy to scale up and operate this system. More summarized data about the advantages and disadvantages of each process for mAb production can be seen in Table 2.
\nProduction platform | \nBatch | \nFeed-batch | \nPerfusion culture | \n
---|---|---|---|
Advantages | \n
| \n
| \n
| \n
Disadvantages | \n
| \n
| \n
| \n
A lot of effort has been made to increase cell longevity in batch and feed-batch modes of operation. It is expected that the longer the cells are maintained viable, the greater the antibodies’ production will be. So, in order to maintain cell viability, some culture parameters can be optimized, such as culture media, feed solution, and mAb secretion rates and by-product production. To improve mAb titers in the batch platform, the start medium can be supplemented with glucose and amino acids, increasing mAb production up to eightfold when compared with regular media [9, 26]. Improvements for the fed-batch platform can be achieved by adjustments in feed solution, as mentioned before. Feed solutions containing glucose and aminoacids/glutamine have been reported to increase mAb titers from two to fourfold, reaching production of up to 2 g/L, when compared with the batch production platform [19].
\nThe optimization of the antibody secretion rate can be achieved by high-density cell cultivation. On a fed-batch platform, a high cell cultivation culture can reach an mAb productivity rate of 0.94 g/L/day and a final titration of 17 g/L, while a continuous culture performed at high density conditions can reach final titration and productivity rates of 0.8 and 1.6 g/L/day, respectively [20]. Optimizing mAb secretion highly depends on the cell line chosen for production. Each cell strain can be influenced by the manufacturing conditions and respond differently to increasing or decreasing mAb production and secretion [19]. The accumulation of toxic by-products is a great bottleneck in manufacturing processes since they can inhibit cell growth and then directly affect mAb production. Although a few strategies to minimize this by-product accumulation have shown to be promising, some are not applicable for a large-scale production. Optimizing medium composition and feed solutions with substrates that reduce toxic compound production is the most common strategy used at industrial scales of production [19].
\nAlthough most mAbs are produced by fed-batch process, there are tendencies indicating that in the future many bioprocesses will be operated in continuous platforms, especially for the production of biopharmaceuticals. On these platforms, the production system will be coupled to upstream and downstream processes [21]. However, for this to actually happen, a great improvement in technological development still needs to be achieved.
\nThe use of monoclonal antibodies as therapeutic drugs requires a large-scale production that far exceeds that of laboratory production (Figure 2). Various production systems have been developed and have evolved, while new alternatives are emerging. The production of mAbs at commercial scale can be performed with adherent cells or suspension cells, although the latter is by far the most used and is better established with more efficient production methods available for cells cultivation. Thus, scale-up using suspension cells is easier. Another advantage of the suspension production system is that a bioreactor with a large area for cell adhesion is not necessary since the cultivation of adherent cell productivity is directly linked to the bioreactor’s area [22].
\nWork volumes used for industrial production of some commercial monoclonal antibodies [
Some cultivation issues and worries have arisen regarding the production scale increase, maintenance of product quality, contamination control, demand for oxygen supply, and control over DO and CO2 removal, among others. Regarding suspension cell cultures, aeration is in part dependent on the agitation of the culture inside the bioreactor, which can lead to cell shear stress. To work around cultivation problems, major advances have been made in the process itself by developing better culture control and conditions, as well as the improvement and development of new bioreactors [7, 23].
\nThe different types of bioreactors commonly used for mAb production in submerged mammalian cells are stainless steel stirred tank bioreactors (STR), air-lift reactors, and disposable bioreactors. More details on each of these bioreactors are discussed below.
\nSchematic representation of a stainless steel stirred tank bioreactor. Showing the main components in a cell cultivation.
Stainless steel stirred tank bioreactors are the most consolidated type of bioreactor used for industrial mAb production and consist of baffle-stirred tanks linked to rotor systems (Figure 3). It is a consolidated system, and there is a lot of knowledge and experience surrounding this technology, acquired by its vast industrial use beyond production using mammalian cells.
\nThe cultivation in this bioreactor allows for wide flexibility of working volumes, ranging from 1.0 to 25.0 L [1], since this system is easily scalable to larger volumes due to its high control over production conditions and extensive handling knowledge. The mechanisms and cleaning and sterilization protocols are well defined. Additionally, cultivation parameters for this system, such as gas transfer coefficient, agitation, aeration, temperature maintenance, pH, and others are well controlled and regulated when compared to other production systems. Another advantage of the STR is that it can be used for cultivation of various cell types and in addition, the products obtained from the cultivation in this type of bioreactor are easily approved for therapeutic use, as regulatory terms are well defined for this type of production [11].
\nHowever, the biggest disadvantage for the use of STR is the stress caused by shear. It can cause cell lysis and lead to loss in mAb productivity.
\nSchematic representation of an air-lift bioreactor. Showing the main components in a cultivation process.
Air-lift reactors are also broadly used for the industrial production of mAbs. The reactor consists of tanks with a bubble column inside, and air is injected into the column base (Figure 4). The air flows through the column’s length to the top of the bioreactor as degassed culture medium flows in the opposite direction to the reactor bottom. This creates a constant gentle mixing of the medium as well as proper culture aeration, annulling part of the shear stress caused by other stirring systems. Other advantages of this operation system are that it is easier to scale-up, contamination problems are more unlikely to occur, and the equipment is simpler. In spite of these advantages, this system is less utilized than STR reactors because the working volume ranges only from 2.0 to 5.0 L [1] and the air-lift reactor handling is not so well elucidated [11].
\nThe first single-use bioreactors emerged in the late 1990s with the launch of a wave reactor system. After that, disposable stirred tank bioreactors were developed [11].
\nThis method brought many advantages for mAb manufacturing. At the end of the process, the bioreactor is discarded and replaced by a new clean and sterile one. This eradicates cross contamination between batches and decreases the time consumed with the equipment preparation between batches. When all the advantages of this process are taken in account, the savings made regarding production and investment capital are highly significant when compared with other process methods. The great disadvantage of this production system is the small work volume supported, ranging from 50 to 2000 L [1].
\nThe wave system consists of a sterile plastic bag (CellBag™) lying on a rocking platform (Figure 5). The bag is half filled with cultivation medium and half filled with a gas mix of interest. The platform motion creates an undulation movement in the culture, ensuring efficient aeration and culture mixing without causing shear damage [10, 11, 13]. The other available systems combine the convenience of a disposable system with the well-known stirred tank system and they are HyClone S.U.B®, Millipore® (CellReady™), or Xcellerex® (XDR™).
\nSchematic representation of a disposable wave bioreactor. Showing the main components in a cell cultivation process.
The main features of SUBs are related to their technical characteristics similar to those of stainless steel bioreactors, that is, aeration rate, agitation, reactor geometry, and ease of monitoring internal conditions, a process similar to stainless steel bioreactors [9].
\nSUBs are being widely used to replace many processes for the production of bioproducts. SUBs may be a cheaper and more efficient alternative from an industrial point of view, and its principle can easily replace any bioprocess to adapt the method to the platform of interest to be replaced, such as large tanks and stainless steel or the motion rocking platforms [9, 24].
\nSUBs have been used in bioprocesses for monoclonal antibody production involving several expression systems, including mammalian cells, microorganisms, plants, mammary glands, etc. Animal cell culture technology is one of the oldest techniques for the production of mAbs.
\nThere is also the production of bottles known as roller bottles, consisting of mammalian cells growing in nutritional and physical conditions controlled in bottles which remain in rotational movement.
\nRoller bottles are a rotary motion system for growing cells and for the production of some bioproducts. It has been an alternative to other monoclonal antibody production systems (Figure 6). Roller bottles provide conditions that favor the transfer of oxygen and temperature control without aeration, agitation propellers, or circulation pumps. The bottle is mounted on a turntable which gives homogeneity of growth and aeration of the culture medium [11, 25, 28].
\nSchematic representation of roller bottles bioreactor and a rack with the rotational motion system in a cultivation for mAb production.
For the production of monoclonal antibodies at commercial scale, the roller bottle technique can be adapted to racks containing tens of bottle in a production line. The advantages of this technique is the high growth potential linked to ease of handling and monitoring of certain conditions such as temperature and rotation. However, the scale of view requires a large physical footprint, which can make the process less economical [11, 25].
\nActually, the trade of monoclonal antibodies makes up half of marketed biopharmaceuticals, reaching $ 75 billion. For some years, new development methodologies of antibodies have advanced with new production processes, allowing scale-up production and market introduction, and demands for high-quality biologics will continue to increase in the coming decades. Generally, processes are similar to those applied in the scheduling for other bioproducts/biosimilars that rely on culturing microorganisms or cells, requiring the development of a well-designed culturing process comprising the full range of control and associated operations that will support technical evaluations.
\nIn combination with increasing pressure from regulatory agencies for enhanced quality and lower process costs from the health care systems, we are facing an important challenge. It will be necessary to make changes in plant design aiming for highly flexible multi-purpose facilities for small production volumes.
\nMicroRNAs or miRNAs are a class of small non-coding RNA approximately 21–25 nucleotides that modulate on gene expression post-transcriptionally via binding to the 3′ untranslated region (3′-UTR) of the target messenger RNA (mRNA), resulting in mRNA degradation or translational repression. The first miRNA, lin-4, was discovered by Ambro and his research group in 1993 and it was found to be related with larva development in
miRNAs are normally transcribed by RNA polymerase II from miRNA genes. This transcription leads to generate a primary miRNA transcript (pri-miRNA). Then, pri-miRNA is further cleaved by a microprocessor complex, which consists of Drosha, the double-stranded RNase III enzyme and DiGeorge syndrome critical region 8 (DGCR8), important cofactor, into a hairpin structure precursor miRNA (pre-miRNA) in the nucleus (Figure 1). The double strand pre-miRNAs with 70 nucleotides are then exported to the cytoplasm by the process of nuclear export factor exportin-5. The pre-miRNA is then processed by RNase III, Dicer, thereby generating a mature miRNA:miRNA duplex approximately 22 nucleotides in length and without a hairpin structure. The helicase enzyme cleaves miRNA duplexes into single-stranded miRNAs and incorporated into the Argonaute (AGO), TRBP and PACT proteins to form the RNA-induced silencing complex (RISC). Usually, other single strand called passenger strand or the star (*) strand will be degraded, while single strand mature miRNA is able to bind with its target mRNA and mediating translational inhibition or mRNA degradation, along with their sequence complementarity to the target [1, 3]. In plants, target mRNA will be degraded if miRNA has perfect or near-perfect complementarity to its target. In contrast to mammal, miRNAs bind to partially complementary sites in the 3′-UTRs of target mRNA, which leading to translational repression [4]. the target mRNA is either blocked (imperfect complementary) or degraded (perfect complementary) of the ribosomal translation, which sequentially impacts the cellular functions.
miRNA biogenesis. miRNA gene is transcribed by RNA polymerase II and then forming the primary miRNA transcript (pri-miRNA), which is further cleaved by the Drosha/DGCR8 complex to generate the precursor miRNA (pre-miRNA). Pre-miRNA is then exported into the cytoplasm by exportin 5/RAN-GTP and further processed by dicer to create the mature miRNA, which is loaded into RISC, which contains AGO, PACT and TRBP proteins. Mature miRNA that binding to its target mRNA by perfect complementary binding and resulting in gene suppression by mRNA degradation. The partially complementary binding of miRNA and its target mRNA, which in turn inhibit the protein translation.
Phytochemicals are major plant-derived compounds that naturally found in vegetables, fruits, medicinal plants or other plants with medicinal properties including antioxidant, anti-diabetic, anti-inflammatory, antimicrobial, antidepressant, anticancer and prevention in other chronic non-communicable diseases [5, 6, 7]. Phenolic and flavonoid compounds are the most important group of bioactive compounds and second metabolites in plants which comprise of essential molecules of human diet [6, 8]. It has been shown that bioactive compounds can modulate the endogenous miRNAs expression [1, 9, 10, 11, 12]. Recently, some studies have revealed that plant-derived miRNAs (dietary miRNAs) as new bioactive compounds in plants can affect the synthesis of endogenous miRNAs [13, 14, 15]. Strikingly, miRNAs do not function only their origins but they are able to regulate the gene expression in cross-kingdom. Therefore, bioactive compounds present in functional foods are potentially regulate endogenous miRNAs expression.
Extensive studies have been performed to understand the molecular mechanism of bioactive compounds with a positive effect on chronic diseases or non-communicable diseases such as arthritis, cancer, cardiovascular diseases, diabetes and obesity [1, 16]. Emerging evidences confirm that alteration of endogenous miRNAs expression can be influenced by bioactive compounds in functional foods [16, 17] (Figure 2 and Table 1).
Influences of bioactive compounds and dietary miRNAs on human non-communicable diseases. Ascending arrows represent up-regulated miRNAs and descending arrows represent down-regulated miRNAs by bioactive compounds. The green triangles show the positive impact of dietary miRNAs on human health.
Dietary compound | miRNA expression | Target of miRNA | Diseases | References | |
---|---|---|---|---|---|
Up-regulation | Down-regulation | ||||
Acetyl-11-Keto-β-Boswellic Acid | miR-27a miR-34a | Unknown | Colorectal cancer | [23] | |
miR-155 | SOCS-1 | Neuroinflammation | [21] | ||
miR-206 | ER-α | Breast cancer | [22] | ||
Arctigenin | miR-16 miR-199a | Unknown | Neuroinflammation | [28] | |
miR-21 miR-19b miR-148a | Unknown | Prostate cancer | [29] | ||
Cinnamic acid derivatives | miR-143 | MAPK/Erk5 | Colon cancer | [31] | |
miR-145 | Unknown | Gastric cancer | [33] | ||
Curcumin | miR-15a, miR-16, miR-34a, miR-146b-5p miR-181b | miR-19a miR-19b | Unknown | Breast cancer | [38] |
miR-101, miR-200b, miR-200c, miR-141 miR-429 | miR-21 | Unknown | Colorectal cancer | [39, 40] | |
miR-21 | Gastric cancer | [41] | |||
miR-145 miR-1275 miR-1908 miR-3127 miR-3178 miR-3198 | miR-23b*, miR-183 miR-193b* miR-210 miR-222* miR-494 miR-664* miR-671-5p | Oct4 | Prostate cancer | [42] | |
miR-181b | CXCL1 CXCL2 | Breast cancer | [43] | ||
miR-378 | p38 | glioblastoma | [44] | ||
miR-124 miR-155 | Unknown | Neurodegenerative disorder | [45] | ||
3,3′-Diindolyl-methane | let-7 miR-34a miR-150-5p | EZH2, Notch1 AR Ahr | Prostate cancer | [46] | |
miR-200 | FoxM1 | Breast cancer | [47] | ||
miR-212/132 cluster miR-21 | Sox4 Cdc25A | Breast cancer | [48, 49] | ||
let-7b, let-7c, let-7d, let-7e, and miR-200b/c | ZEB-1, E-cadherin | Pancreatic cancer | [50] | ||
miR-146a | Unknown | Pancreatic cancer | [51] | ||
(−)-Epigallocatechin-3-Gallate | miR-296 | STAT3 | Nasopharyngeal carcinoma | [57] | |
let-7a miR34a | c-Myc | Hepatocellular carcinoma | [58] | ||
miR-34a | miR-93 | Unknown | Prostate cancer | [59] | |
miR-29 miR-210 | miR-125b miR-203 | Unknown | Cervical cancer | [60] | |
let-7 | HMGA2 | Melanoma cell | [61] | ||
miR-384 | Beclin-1 | Myocardial ischemia/ reperfusion | [62] | ||
miR-140-3p | Unknown | Osteoarthritis | [63] | ||
miR-10b miR-181a miR-221 | Unknown | Liver fibrosis | [64] | ||
Genistein | miR-23b | Unknown | Breast cancer | [66] | |
miR-1260b | sRRP1 Smad4 | Prostate cancer | [67] | ||
miR-1260b | sFRP1, Dkk2, Smad4 | Renal cancer | [68] | ||
miR-27a | Unknown | Lung cancer | [69] | ||
miR-29b | Unknown | Lung cancer | [70] | ||
miR-451 | Unknown | Chronic liver disease | [72] | ||
Quercetin | miR-200b-3p | Notch1 | Pancreatic cancer | [75] | |
miR-146a | EGFR | Breast cancer | [76] | ||
miR-16 | HOXA10 | Oral cancer | [77] | ||
miR-22 | WNT1/β-catenin | Oral cancer | [78] | ||
miR-97 miR-298 miR-2218 miR-1502 miR-2117 | Unknown | Oxidative stress in pheochromocytoma | [79] | ||
miR-503-5p miR-1283, miR-3714 miR-6867-5p | CCND1 | Endometriosis | [80] | ||
miR-122 | miR-21 | Unknown | Liver fibrosis | [81] | |
miR-199 | Sert1 | Hypoxia | [82] | ||
Silymarin | miR-203 | class 1 HDAC proteins and ZEB1 | Lung cancer | [84] | |
miR-155 | Unknown | Rheumatoid arthritis | [85] | ||
miR-122 | Unknown | Liver damage | [86] | ||
miR-122 miR-192 miR-194 | Unknown | Liver damage | [87] | ||
β-Sitosterol-d-glucoside | miR-10a | Unknown | Breast cancer | [89] | |
Sulforaphane | miR-23b miR-92b miR-381 miR-382 | Unknown | Breast cancer | [92] | |
miR-616-5p | GSK3β/β-catenin | Lung cancer | [93] | ||
miR-135b-5p | miR-30a-3p | RASAL2 Cx43 | Pancreatic cancer | [94] | |
miR-200c | Unknown | Oral cancer | [96] | ||
miR-9 miR-326 | Unknown | Gastric cancer | [97] | ||
miR-124-3p | STAT3 | Nasopharyngeal cancer | [98] | ||
miR-423-5p | Unknown | Liver fibrosis | [99] | ||
miR-155 | Unknown | Neuroinflammation | [100] |
Summary of miRNAs bioactive compounds and miRNAs expression in human pathology.
3-acetyl-11-keto- β -boswellic acid (AKBA) is pentacyclic triterpene acids that mainly found in
Arctigenin (AR) is a phenylpropanoid dizbenzylbutyrolactone lignin and was first identified in
Cinnamic acid derivatives can occur naturally in plants and their structure composing of benzene ring and acrylic acid group. Several compounds of cinnamic acid derivatives have been identified including artepilin C, baccharin, drupanin, ferulic acid, curcumin, caffeic acid, p-hydroxycinnamic acid, coumaric and chlorogenic acids, etc. [30, 31]. Medicinal activities of cinnamic acid derivatives have been reported such as anti-inflammatory, anti-oxidant, anti-viral, anti-microbial, anti-diabetic, neuroprotective and anti-tumor activities [30, 31, 32]. Cinnamic acid derivatives from propolis significantly induced colon cancer cell apoptosis through TRAIL/DR4/5 and/or FasL/Fas death-signaling pathways and via the upregulated miR-143 expression, resulting in decreased the target gene MAPK/Erk5 expression and its downstream target c-Myc [31]. Moreover, Li et al. demonstrated that cinnamic acid derivatives decreased gastric cancer cell proliferation through the up-regulation of miR-145 and down-regulation P13K/Akt signaling pathway [33]. Therefore, cinnamic acid derivatives have a potential as therapeutic agents for cancer.
Curcumin[(1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptane-3,5-dione] is well known as natural polyphenol and derived from the rhizome of turmeric or
3,3′-diindolylmethane (DIM) is a naturally active compound found in stomach, which derived from indole-3-carbinol (I3C) that present in cruciferous vegetables. DIM has been reported to regulate several miRNAs expression in cancer. Tumor suppressor miRNAs was upregulated by DIM in prostate cancer cells including let-7, miR-34a and miR-150-5p by targeting EZH2, Notch1 and AR and Ahr, respectively [46]. DIM also upregulated tumor suppressor miR-200, which led to inhibit the expression of FoxM1 in breast cancer cells [47]. miR-212/132 cluster and miR-21 were upregulated by DIM, which downregulated the expression of Sox4 and Cdc25A, respectively in breast cancer [48, 49]. Moreover, DIM upregulated let-7b, let-7c, let-7d, let-7e, and miR-200b/c expression, which led to inhibit the expression of ZEB-1, E-cadherin in pancreatic cancer cells [50]. It has been reported miR-146a was upregulated upon treated with DIM and suppressed the expression of MTA2, NF-κB, IRAK1, EGFR in pancreatic cancer cells [51].
DIM showed the modulation of miRNAs expression in other inflammatory diseases. The expression of miR-106a, miR-20b, and miR-125b-5p were increased after treatment with DIM and suppressed the expression of IRAK4 and TNF-α to limit responses to TLRs activated by LPS in acute liver failure (ALF) animal model [52]. DIM significantly upregulated miR-200c, miR-146a, miR-16, miR-93, and miR-22 in brain CD4+ T cells and inhibited the expression of cyclin E1 and B-cell lymphoma-2 in experimental autoimmune encephalomyelitis (EAE), a murine model of multiple sclerosis [53].
(−)-Epigallocatechin-3-Gallate or EGCG is a major polyphenol compound in green tea (
EGCG showed the protective effect against myocardial ischemia/reperfusion (I/R) injury through up-regulation of miR-384-mediated autophagy by targeting Beclin-1 via activating the PI3K/Akt signaling pathway [62]. EGCG also demonstrated the anti-arthritic effects by inhibited IL-1β-induced ADAMTS5 expression and up-regulated the expression of miR-140-3p in osteoarthritis chondrocytes [63]. EGCG treatment has potential role of preventing toxin-induced fibrosis by suppression of osteopontin expression and up-regulation of miR-10b, miR-181a and miR-221 in liver hepatocellular carcinoma cells [64].
Genistein belongs to isoflavone family and presents in soybeans with antiangiogenic, anti-metastasis, anti-inflammatory, anti-oxidant, cell cycle arrest and induction of apoptosis effects [65]. Genistein can regulate the expression of miRNAs in several call types [65]. It has been reported that treatment of genistein up-regulated miR-23b and inhibited breast cancer cell growth [66]. Genistein also exhibited anti-tumor effect by down-regulated miR-1260b and targeting sRRP1 and Smad4 through DNA methylation or histone modifications in prostate cancer cells [67]. The same research group reported that miR-1260b was highly expressed in renal cancer cells and miR-1260b was down-regulated in genistein treated renal cancer cells [68]. The treatment of miR-1260b inhibitor inhibited the expression of its target genes, sFRP1, Dkk2, Smad4 [68]. Treatment with genistein induced non-small lung cancer cell apoptosis, caspase-3/9 activation and inhibited cell proliferation via up-regulation of miR-27a -mediated MET signaling [69]. Co-encapsulate miR-29b with genistein in hybrid nanoparticles (GMLHN) has been studied to treat effectively in non-small lung cancer cell and GMLHN showed the anti-proliferative effect by down-regulation of phosphorylated AKT (pAKT) and phosphorylated phosphoinositide-3 kinase (p-PI3K) [70].
Genistein promoted myoblast proliferation and differentiation through down-regulated miR-222 expression, resulting in increased expression of its target genes, MyoG, MyoD, and ERα [71]. Interestingly, genistein up-regulated miR-451 expression and inhibited IL1β expression and inflammation in chronic liver disease nonalcoholic steatohepatitis (NASH) mice model [72].
Quercetin is bioactive flavonoids that can be found in fruits and vegetables including onion, kale, apple, many berries, citrus fruits and tea [73]. Anti-cancer, anti-inflammatory, antioxidant, anti-diabetes, anti-atherosclerosis and anti-viral effects have been reported in different in vitro studies for quercetin [74]. Several studies have focused on quercetin and miRNAs modulation for therapeutic approaches. miR-200b-3p was up-regulated in pancreatic cancer cells when treated with quercetin, resulting in inhibition of self-renewal and decrease of proliferation through Notch1 signaling pathway [75]. Quercetin significantly inhibited breast cancer cell proliferation and invasion via up-regulated miR-146a expression and targeting EGFR [76]. Quercetin inhibited cell viability, migration and invasion by up-regulated miR-16 and targeting HOXA10 in oral cancer cells [77]. In addition, quercetin decreased oral cancer cell viability and increased cell apoptosis via miR-22/WNT1/β-catenin pathway [78].
Recently, quercetin modulated 34 miRNAs expression (5 upregulated and 29 downregulated) and novel miR-97, miR-298, miR-2218, miR-1502, and miR-2117 were identified in pheochromocytoma of the rat adrenal medulla that responded for protective effect against oxidative stress through PI3K-AKT signaling pathway [79]. Treatment of quercetin inhibited proliferation of endometriosis through up-regulated miR-503-5p, miR-1283, miR-3714 and miR-6867-5p by targeting CCND1 [80]. TGFβ1 is a fibrosis inducer and quercetin significantly down-regulated miR-21 and TGFβ1 and up-regulated miR-122 in liver fibrosis [81]. Protection of cardiomyocyte against hypoxia caused insults of quercetin has been reported by up-regulation of miR-199 mediated sirt1 expression and AMPK phosphorylation [82].
Silymarin is a flavonolignans extracted from the milk thistle
β-Sitosterol-d-glucoside is bioactive compounds that has been isolated from
Sulforaphane is dietary compounds in broccoli (
Sulforaphane has potential to inhibit hepatic fibrosis by downregulating miR-423-5p in hepatic stellate cell [99]. Sulforaphane showed the protective effect in microglia-mediated neurotoxicity by inhibited LPS-induced expression of inflammatory miRNA, miR-155 [100].
Several evidences demonstrated the direct modulation of cellular signaling pathways by dietary compounds could decrease the risk of chronic diseases [101]. Interestingly, it has been reported that small non-coding RNA including miRNAs can be transferred across Kingdoms, for example dietary miRNAs have been found in human body fluids and these circulating miRNAs are likely to regulate human gene expression [15, 102, 103, 104, 105, 106, 107]. The uptake of plant derived miRNAs could be in the form of raw and cooked plants in capable of stability forms [107, 108]. Due to high temperature cooking process, low pH and enzymes in digestive tract as well as enzymes in blood circulation, miRNAs might be destroyed before their functions with target mRNAs [15]. Strikingly, GC base content, 2’-O-methylation on the 3′-terminal, unique nucleotide sequence of dietary miRNAs and extracellular vesicles (exosome and microvesicle) are preventive features of plant derived miRNAs in harmful conditions [109, 110, 111, 112, 113, 114].
There are numerous studies to support the functional roles of dietary miRNAs in cross kingdom gene regulation. Rice miR156a and miR168a were detected in human serum and miR168a down-regulated low-density lipoprotein receptor adapter protein 1 (LDLRAP1) expression, resulted in an increase of plasma LDL cholesterol level, Table 2 [105]. miR2910 from
Plants | Plant derived-miRNAs | Human target gene/ Disease | References |
---|---|---|---|
osa-miR156a osa-miR166a osa-miR168a | LDLRAP1 | [105] | |
peu-miR2910 | JAK–STAT pathway | [115] | |
14 miRNAs | Cancer (breast, lung and leukemia) | [116] | |
miR156 miR531 miR160 miR529b miR1118 | Ras-MAPK signaling pathway, Alzheimer disease, breast cancer, cardiomyopathy, HIV, lung cancer, several neurological disorders | [117] | |
miR14 | Rheumatoid arthritis | [120] | |
cabbage, spinach and lettuce | miR156a | Cardiovascular disease | [118] |
miR156-5p miR164-5p miR168-5p miR395-3p miR396-3p miR396-5p miR444-3p miR529-3p miR1846-3p miR2907-3p | Cancer, cardiovascular and neurodegenerative diseases | [119] |
Dietary miRNAs and human gene regulation.
The abundantly expressed miRNA in dietary green vegetable, miR156a which was detected in human serum and targeted the junction adhesion molecule-A (JAM-A) [118]. The JAM-A was up-regulated in atherosclerotic lesions from cardiovascular disease patients and miR156a could suppressed inflammatory cytokine-induced monocytes adhesion by targeting JAM-A [118]. The very recently report using a computational approach to predict the potential target of rice miRNAs including miR156-5p, miR164-5p, miR168-5p, miR395-3p, miR396-3p, miR396-5p, miR444-3p, miR529-3p, miR1846-3p, miR2907-3p, which can bind to the human mRNA [119]. Most of these target genes were associated with cancer, cardiovascular and neurodegenerative diseases [119]. miR14 derived from
It has been widely known that functional foods and their bioactive compounds have the capacity for human health benefits. To date, miRNAs have been shown a significant effect on gene expression and modulate the cellular biological functions in physiological and pathological conditions. There is emerging evidence suggesting that dietary bioactive compounds can be effective in human diseases as a result of altering miRNAs expression levels, resulting in modulation of cellular signaling pathway. Additional research the possibility of bioactive compounds for developing as novel drugs with less side effects is required
This study has been supported by University of Phayao Research Grant (Grant no. FF64-RIM037) and School of Medical Sciences Research Grant (Grant no. MS 632001), University of Phayao.
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",metaTitle:"Copyright Policy",metaDescription:"Copyright is the term used to describe the rights related to the publication and distribution of original works. Most importantly from a publisher's perspective, copyright governs how authors, publishers and the general public can use, publish and distribute publications.",metaKeywords:null,canonicalURL:"/page/copyright-policy",contentRaw:'[{"type":"htmlEditorComponent","content":"Copyright is the term used to describe the rights related to the publication and distribution of original Works. Most importantly from a publisher's perspective, copyright governs how Authors, publishers and the general public can use, publish, and distribute publications.
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\\n\\nPolicy last updated: 2016-06-08
\\n"}]'},components:[{type:"htmlEditorComponent",content:'Copyright is the term used to describe the rights related to the publication and distribution of original Works. Most importantly from a publisher's perspective, copyright governs how Authors, publishers and the general public can use, publish, and distribute publications.
\n\nIntechOpen only publishes manuscripts for which it has publishing rights. This is governed by a publication agreement between the Author and IntechOpen. This agreement is accepted by the Author when the manuscript is submitted and deals with both the rights of the publisher and Author, as well as any obligations concerning a particular manuscript. However, in accepting this agreement, Authors continue to retain significant rights to use and share their publications.
\n\nHOW COPYRIGHT WORKS WITH OPEN ACCESS LICENSES?
\n\nAgreement samples are listed here for the convenience of prospective Authors:
\n\nDEFINITIONS
\n\nThe following definitions apply in this Copyright Policy:
\n\nAuthor - in order to be identified as an Author, three criteria must be met: (i) Substantial contribution to the conception or design of the Work, or the acquisition, analysis, or interpretation of data for the Work; (ii) Participation in drafting or revising the Work; (iii) Approval of the final version of the Work to be published.
\n\nWork - a Chapter, including Conference Papers, a Scientific Article and any and all text, graphics, images and/or other materials forming part of or accompanying the Chapter/Conference Paper.
\n\nMonograph/Compacts - a full manuscript usually written by a single Author, including any and all text, graphics, images and/or other materials.
\n\nCompilation - a collection of Works distributed in a Book that IntechOpen has selected, and for which the coordination of the preparation, arrangement and publication has been the responsibility of IntechOpen. Any Work included is accepted in its entirety in unmodified form and is published with one or more other contributions, each constituting a separate and independent Work, but which together are assembled into a collective whole.
\n\nScientific Journal – Periodical publication intended to further the progress of science.
\n\nJournal Article/Scientific Article – Publication based on empirical evidence. It can support a hypothesis with original research, describe existing research or comment on current trends in a specific field.
\n\nIntechOpen - Registered publisher with office at 5 Princes Gate Court, London, SW7 2QJ - UNITED KINGDOM
\n\nIntechOpen platform - IntechOpen website www.intechopen.com whose main purpose is to host Monographs in the format of Book Chapters, Long Form Monographs, Compacts, Conference Proceedings, Scientific Journals and Videos.
\n\nVideo Lecture – an audiovisual recording of a lecture or a speech given by a Lecturer, recorded, edited, owned and published by IntechOpen.
\n\nTERMS
\n\nAll Works published on the IntechOpen platform and in print are licensed under a Creative Commons Attribution 3.0 Unported and Creative Commons 4.0 International License, a license which allows for the broadest possible reuse of published material.
\n\nCopyright on the individual Works belongs to the specific Author, subject to an agreement with IntechOpen. The Creative Common license is granted to all others to:
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\n\t\t\t Creative Commons Attribution-NonCommercial-ShareAlike 3.0 Unported (CC BY-NC-SA 3.0) \n\t\t\t | \n\t\t\t1 July 2005 (2005-07-01) | \n\t\t\t3 October 2011 (2011-10-03) | \n\t\t
\n\t\t\t Creative Commons Attribution 3.0 Unported (CC BY 3.0) \n\t\t\t | \n\t\t\t5 October 2011 (2011-10-05) | \n\t\t\tCurrently | \n\t\t
\n\t\t\t Creative Commons 4.0 International (CC BY 4.0) – for Journal Articles \n\t\t\t | \n\t\t\t15 March 2022 | \n\t\t\tCurrently | \n\t\t
The CC BY 3.0 and CC BY 4.0 license permits Works to be freely shared in any medium or format, as well as the reuse and adaptation of the original contents of Works (e.g. figures and tables created by the Authors), as long as the source Work is cited and its Authors are acknowledged in the following manner:
\n\nContent reuse:
\n\n© {year} {authors' full names}. Originally published in {short citation} under {license version} license. Available from: {DOI}
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\n\nReposting & sharing:
\n\nOriginally published in {full citation}. Available from: {DOI}
\n\nRepublishing – More about Attribution Policy can be found here.
\n\nThe same principles apply to Works published under the CC BY-NC-SA 3.0 license, with the caveats that (1) the content may not be used for commercial purposes, and (2) derivative works building on this content must be distributed under the same license. The restrictions contained in these license terms may, however, be waived by the copyright holder(s). Users wishing to circumvent any of the license terms are required to obtain explicit permission to do so from the copyright holder(s).
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\n\nAll rights to Books and Journals and all other compilations published on the IntechOpen platform and in print are reserved by IntechOpen.
\n\nThe copyright to Books, Journals and other compilations is subject to separate copyright from those that exist in the included Works.
\n\nAll Long Form Monographs/Compacts are licensed under the Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0) license granted to all others.
\n\nCopyright to the individual Works (Chapters) belongs to their specific Authors, subject to an agreement with IntechOpen and the Creative Common license granted to all others to:
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\n\nContent reuse:
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\n\nPolicy last updated: 2016-06-08
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Therefore, the selection of proper type of central air conditioning system is a crucial target in the construction industry as improper selection can maximize initial and/or running costs of the system and decreases the human comfort and indoor air quality levels. In fact, a pre-assessment of the construction type and budget available is required for selecting the proper type of central air conditioning system. Therefore, there is a continuous need for an updated material in the literature that reviews the central air conditioning systems and applications, which is the motivation of the present chapter. The present chapter reviews the central air conditioning systems and applications. Specifically, all-air systems, all-water systems, and air-water systems are discussed. In addition, all provided systems are further explored through several developed schematic diagrams enabling the identification of their various components and the understanding of their working principles. It is may be of interest to note that this chapter is suitable for undergraduate level students in the fields of HVAC and R, mechanical, and construction engineering.",book:{id:"8394",slug:"low-temperature-technologies",title:"Low-temperature Technologies",fullTitle:"Low-temperature Technologies"},signatures:"Mohamed Elnaggar and Mohammed Alnahhal",authors:[{id:"178453",title:"Dr.",name:"Mohamed",middleName:null,surname:"Elnaggar",slug:"mohamed-elnaggar",fullName:"Mohamed Elnaggar"},{id:"308344",title:"Dr.",name:"Mohammed",middleName:null,surname:"Alnahhal",slug:"mohammed-alnahhal",fullName:"Mohammed Alnahhal"}]},{id:"69746",title:"Energy and Exergy Analysis of Refrigeration Systems",slug:"energy-and-exergy-analysis-of-refrigeration-systems",totalDownloads:1538,totalCrossrefCites:0,totalDimensionsCites:5,abstract:"Refrigeration systems have the priority in design for residential and industrial applications. The chapter includes five major refrigeration systems: vapor-compression refrigeration; ammonia-water absorption refrigeration; gas refrigeration where standard air is the most popular refrigerant; multi-pressure refrigeration including multistage, cascade, and multipurpose refrigeration system; and heat pump systems. Energy and exergy analysis has been presented for most of the systems. The energetic and the exergetic COP for each system are presented. Renewable energy sources are also discussed including geothermal, solar, and wind energy, a with combination with refrigeration systems in different industrial and residential applications. The overall efficiency of the renewable systems is achieved to be more than 50% providing promising solutions for energy use and having a low environmental impact.",book:{id:"8394",slug:"low-temperature-technologies",title:"Low-temperature Technologies",fullTitle:"Low-temperature Technologies"},signatures:"Shaimaa Seyam",authors:[{id:"257733",title:"MSc.",name:"Shaimaa",middleName:null,surname:"Seyam",slug:"shaimaa-seyam",fullName:"Shaimaa Seyam"}]},{id:"60220",title:"Application of Exergy Analysis to Energy Systems",slug:"application-of-exergy-analysis-to-energy-systems",totalDownloads:2446,totalCrossrefCites:10,totalDimensionsCites:13,abstract:"Exergy analysis is a practical approach to evaluate the merit of energy conversion or distribution processes and systems. With the aid of an energy analysis, the performance of an energy conversion system cannot be evaluated efficiently and precisely. But, an exergy analysis complements and enhances an energy analysis. Exergy analysis involves the application of exergy concepts, balances, and efficiencies to evaluate and improve energy and other systems. Many scientists suggest that processes or sytems can be well evaluated and improved using exergy analysis in addition to or in place of energy analysis. Application of exergy analysis has given us more beneficial opportunities through a big part of a wide range of processes and systems particularly for the evaluation of energy systems and technologies as well as an environmental impact in all existing thermal and nuclear power plants. Conventional energy technologies, especially for power generation plants, have made numerous energy and exergy analyses and have produced beneficial results. Also, the use of energy and exergy analyses for advanced nuclear energy technologies can be expected to provide meaningful insights into performance that can assist in achieving optimal design concepts. Finally, explaining the analysis of thermal and nuclear power plant systems deals with exergetic approach.",book:{id:"6469",slug:"application-of-exergy",title:"Application of Exergy",fullTitle:"Application of Exergy"},signatures:"Rauf Terzi",authors:[{id:"222042",title:"Dr.",name:"Rauf",middleName:null,surname:"Terzi",slug:"rauf-terzi",fullName:"Rauf Terzi"}]},{id:"68598",title:"Impact of Air-Conditioning Filters on Microbial Growth and Indoor Air Pollution",slug:"impact-of-air-conditioning-filters-on-microbial-growth-and-indoor-air-pollution",totalDownloads:1771,totalCrossrefCites:3,totalDimensionsCites:7,abstract:"Contemporary lifestyles dictate that people spend between 60 and 90% of their daily lives indoors. For those living in warm climates, air conditioning is thus considered a necessity. Air conditioners function by removing hot and humid air from building interior and replacing it with cooler air. Microorganisms are considered among the most important sources of poor quality of indoor air, and contamination of this air by microbial pollutants is being increasingly recognized as a public health problem and a probable cause of the so-called sick building syndrome. In this regard, microfiber glass panel filters are considered to provide an effective solution for air filtration and have been demonstrated to improve air quality in many applications. However, recent research has demonstrated that certain microorganisms are able to colonize panel filter surfaces. Studies on selected microbes isolated from the most commonly used filters have revealed that the bacterial and fungal moist masses carried on sponge-type filters are greater than those carried on polyester and high-efficiency particulate air (HEPA) filters. Moreover, microbial moist mass has been found to increase with increasing incubation time. In addition, recent research has shown that certain microorganisms, particularly fungi, can colonize the materials used in heating, ventilation, and air-conditioning systems (HVAC).",book:{id:"8394",slug:"low-temperature-technologies",title:"Low-temperature Technologies",fullTitle:"Low-temperature Technologies"},signatures:"Amira Hassan Al-abdalall, Sarah Abdullah Al-dakheel and Hmidah Abdulhadi Al-Abkari",authors:[{id:"301509",title:"Dr.",name:"Amira",middleName:null,surname:"A-Abdalall",slug:"amira-a-abdalall",fullName:"Amira A-Abdalall"}]},{id:"45134",title:"Fouling in Membrane Filtration and Remediation Methods",slug:"fouling-in-membrane-filtration-and-remediation-methods",totalDownloads:5781,totalCrossrefCites:27,totalDimensionsCites:68,abstract:null,book:{id:"3072",slug:"mass-transfer-advances-in-sustainable-energy-and-environment-oriented-numerical-modeling",title:"Mass Transfer",fullTitle:"Mass Transfer - Advances in Sustainable Energy and Environment Oriented Numerical Modeling"},signatures:"A. 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Much of biochemistry is devoted to enzymes, proteins that catalyze chemical reactions, enzyme structures, mechanisms of action and their roles within cells. Biochemistry also studies small signaling molecules, coenzymes, inhibitors, vitamins, and hormones, which play roles in life processes. Biochemical experimentation, besides coopting classical chemistry methods, e.g., chromatography, adopted new techniques, e.g., X-ray diffraction, electron microscopy, NMR, radioisotopes, and developed sophisticated microbial genetic tools, e.g., auxotroph mutants and their revertants, fermentation, etc. More recently, biochemistry embraced the ‘big data’ omics systems. Initial biochemical studies have been exclusively analytic: dissecting, purifying, and examining individual components of a biological system; in the apt words of Efraim Racker (1913 –1991), “Don’t waste clean thinking on dirty enzymes.” Today, however, biochemistry is becoming more agglomerative and comprehensive, setting out to integrate and describe entirely particular biological systems. The ‘big data’ metabolomics can define the complement of small molecules, e.g., in a soil or biofilm sample; proteomics can distinguish all the comprising proteins, e.g., serum; metagenomics can identify all the genes in a complex environment, e.g., the bovine rumen. 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Dr. Blumenberg’s research is focused on the epidermis, expression of keratin genes, transcription profiling, keratinocyte differentiation, inflammatory diseases and cancers, and most recently the effects of the microbiome on the skin. He has published more than 100 peer-reviewed research articles and graduated numerous Ph.D. and postdoctoral students.",institutionString:null,institution:{name:"New York University Langone Medical Center",institutionURL:null,country:{name:"United States of America"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:4,paginationItems:[{id:"14",title:"Cell and Molecular Biology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/14.jpg",isOpenForSubmission:!0,editor:{id:"165627",title:"Dr.",name:"Rosa María",middleName:null,surname:"Martínez-Espinosa",slug:"rosa-maria-martinez-espinosa",fullName:"Rosa María Martínez-Espinosa",profilePictureURL:"https://mts.intechopen.com/storage/users/165627/images/system/165627.jpeg",biography:"Dr. Rosa María Martínez-Espinosa has been a Spanish Full Professor since 2020 (Biochemistry and Molecular Biology) and is currently Vice-President of International Relations and Cooperation development and leader of the research group 'Applied Biochemistry” (University of Alicante, Spain). Other positions she has held at the university include Vice-Dean of Master Programs, Vice-Dean of the Degree in Biology and Vice-Dean for Mobility and Enterprise and Engagement at the Faculty of Science (University of Alicante). She received her Bachelor in Biology in 1998 (University of Alicante) and her PhD in 2003 (Biochemistry, University of Alicante). She undertook post-doctoral research at the University of East Anglia (Norwich, U.K. 2004-2005; 2007-2008).\nHer multidisciplinary research focuses on investigating archaea and their potential applications in biotechnology. She has an H-index of 21. She has authored one patent and has published more than 70 indexed papers and around 60 book chapters.\nShe has contributed to more than 150 national and international meetings during the last 15 years. Her research interests include archaea metabolism, enzymes purification and characterization, gene regulation, carotenoids and bioplastics production, antioxidant\ncompounds, waste water treatments, and brines bioremediation.\nRosa María’s other roles include editorial board member for several journals related\nto biochemistry, reviewer for more than 60 journals (biochemistry, molecular biology, biotechnology, chemistry and microbiology) and president of several organizing committees in international meetings related to the N-cycle or respiratory processes.",institutionString:null,institution:{name:"University of Alicante",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null},{id:"15",title:"Chemical Biology",coverUrl:"https://cdn.intechopen.com/series_topics/covers/15.jpg",isOpenForSubmission:!0,editor:{id:"441442",title:"Dr.",name:"Şükrü",middleName:null,surname:"Beydemir",slug:"sukru-beydemir",fullName:"Şükrü Beydemir",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00003GsUoIQAV/Profile_Picture_1634557147521",biography:"Dr. Şükrü Beydemir obtained a BSc in Chemistry in 1995 from Yüzüncü Yıl University, MSc in Biochemistry in 1998, and PhD in Biochemistry in 2002 from Atatürk University, Turkey. He performed post-doctoral studies at Max-Planck Institute, Germany, and University of Florence, Italy in addition to making several scientific visits abroad. He currently works as a Full Professor of Biochemistry in the Faculty of Pharmacy, Anadolu University, Turkey. Dr. Beydemir has published over a hundred scientific papers spanning protein biochemistry, enzymology and medicinal chemistry, reviews, book chapters and presented several conferences to scientists worldwide. He has received numerous publication awards from various international scientific councils. He serves in the Editorial Board of several international journals. Dr. Beydemir is also Rector of Bilecik Şeyh Edebali University, Turkey.",institutionString:null,institution:{name:"Anadolu University",institutionURL:null,country:{name:"Turkey"}}},editorTwo:{id:"13652",title:"Prof.",name:"Deniz",middleName:null,surname:"Ekinci",slug:"deniz-ekinci",fullName:"Deniz Ekinci",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYLT1QAO/Profile_Picture_1634557223079",biography:"Dr. Deniz Ekinci obtained a BSc in Chemistry in 2004, MSc in Biochemistry in 2006, and PhD in Biochemistry in 2009 from Atatürk University, Turkey. He studied at Stetson University, USA, in 2007-2008 and at the Max Planck Institute of Molecular Cell Biology and Genetics, Germany, in 2009-2010. Dr. Ekinci currently works as a Full Professor of Biochemistry in the Faculty of Agriculture and is the Head of the Enzyme and Microbial Biotechnology Division, Ondokuz Mayıs University, Turkey. He is a member of the Turkish Biochemical Society, American Chemical Society, and German Genetics society. Dr. Ekinci published around ninety scientific papers, reviews and book chapters, and presented several conferences to scientists. He has received numerous publication awards from several scientific councils. Dr. Ekinci serves as the Editor in Chief of four international books and is involved in the Editorial Board of several international journals.",institutionString:null,institution:{name:"Ondokuz Mayıs University",institutionURL:null,country:{name:"Turkey"}}},editorThree:null},{id:"17",title:"Metabolism",coverUrl:"https://cdn.intechopen.com/series_topics/covers/17.jpg",isOpenForSubmission:!0,editor:{id:"138626",title:"Dr.",name:"Yannis",middleName:null,surname:"Karamanos",slug:"yannis-karamanos",fullName:"Yannis Karamanos",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002g6Jv2QAE/Profile_Picture_1629356660984",biography:"Yannis Karamanos, born in Greece in 1953, completed his pre-graduate studies at the Université Pierre et Marie Curie, Paris, then his Masters and Doctoral degree at the Université de Lille (1983). He was associate professor at the University of Limoges (1987) before becoming full professor of biochemistry at the Université d’Artois (1996). He worked on the structure-function relationships of glycoconjugates and his main project was the investigations on the biological roles of the de-N-glycosylation enzymes (Endo-N-acetyl-β-D-glucosaminidase and peptide-N4-(N-acetyl-β-glucosaminyl) asparagine amidase). From 2002 he contributes to the understanding of the Blood-brain barrier functioning using proteomics approaches. He has published more than 70 papers. His teaching areas are energy metabolism and regulation, integration and organ specialization and metabolic adaptation.",institutionString:null,institution:{name:"Artois University",institutionURL:null,country:{name:"France"}}},editorTwo:null,editorThree:null},{id:"18",title:"Proteomics",coverUrl:"https://cdn.intechopen.com/series_topics/covers/18.jpg",isOpenForSubmission:!0,editor:{id:"200689",title:"Prof.",name:"Paolo",middleName:null,surname:"Iadarola",slug:"paolo-iadarola",fullName:"Paolo Iadarola",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bSCl8QAG/Profile_Picture_1623568118342",biography:"Paolo Iadarola graduated with a degree in Chemistry from the University of Pavia (Italy) in July 1972. He then worked as an Assistant Professor at the Faculty of Science of the same University until 1984. In 1985, Prof. Iadarola became Associate Professor at the Department of Biology and Biotechnologies of the University of Pavia and retired in October 2017. Since then, he has been working as an Adjunct Professor in the same Department at the University of Pavia. His research activity during the first years was primarily focused on the purification and structural characterization of enzymes from animal and plant sources. During this period, Prof. Iadarola familiarized himself with the conventional techniques used in column chromatography, spectrophotometry, manual Edman degradation, and electrophoresis). Since 1995, he has been working on: i) the determination in biological fluids (serum, urine, bronchoalveolar lavage, sputum) of proteolytic activities involved in the degradation processes of connective tissue matrix, and ii) on the identification of biological markers of lung diseases. In this context, he has developed and validated new methodologies (e.g., Capillary Electrophoresis coupled to Laser-Induced Fluorescence, CE-LIF) whose application enabled him to determine both the amounts of biochemical markers (Desmosines) in urine/serum of patients affected by Chronic Obstructive Pulmonary Disease (COPD) and the activity of proteolytic enzymes (Human Neutrophil Elastase, Cathepsin G, Pseudomonas aeruginosa elastase) in sputa of these patients. More recently, Prof. Iadarola was involved in developing techniques such as two-dimensional electrophoresis coupled to liquid chromatography/mass spectrometry (2DE-LC/MS) for the proteomic analysis of biological fluids aimed at the identification of potential biomarkers of different lung diseases. He is the author of about 150 publications (According to Scopus: H-Index: 23; Total citations: 1568- According to WOS: H-Index: 20; Total Citations: 1296) of peer-reviewed international journals. 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She gained considerable experience in developing and validating new methodologies whose applications allowed her to determine both the amount of biomarkers (Desmosine and Isodesmosine) in the urine of patients affected by COPD, and the activity of proteolytic enzymes (HNE, Cathepsin G, Pseudomonas aeruginosa elastase) in the sputa of these patients. Simona Viglio was also involved in research dealing with the supplementation of amino acids in patients with brain injury and chronic heart failure. She is presently engaged in the development of 2-DE and LC-MS techniques for the study of proteomics in biological fluids. The aim of this research is the identification of potential biomarkers of lung diseases. 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Singh",profilePictureURL:"https://mts.intechopen.com/storage/users/329385/images/system/329385.png",institutionString:"Punjab Technical University",institution:{name:"Punjab Technical University",institutionURL:null,country:{name:"India"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{type:"book",id:"8018",title:"Extracellular Matrix",subtitle:"Developments and Therapeutics",coverURL:"https://cdn.intechopen.com/books/images_new/8018.jpg",slug:"extracellular-matrix-developments-and-therapeutics",publishedDate:"October 27th 2021",editedByType:"Edited by",bookSignature:"Rama Sashank Madhurapantula, Joseph Orgel P.R.O. and Zvi Loewy",hash:"c85e82851e80b40282ff9be99ddf2046",volumeInSeries:23,fullTitle:"Extracellular Matrix - Developments and Therapeutics",editors:[{id:"212416",title:"Dr.",name:"Rama Sashank",middleName:null,surname:"Madhurapantula",slug:"rama-sashank-madhurapantula",fullName:"Rama Sashank Madhurapantula",profilePictureURL:"https://mts.intechopen.com/storage/users/212416/images/system/212416.jpg",institutionString:"Illinois Institute of Technology",institution:{name:"Illinois Institute of Technology",institutionURL:null,country:{name:"United States of America"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null},{type:"book",id:"9759",title:"Vitamin E in Health and Disease",subtitle:"Interactions, Diseases and Health Aspects",coverURL:"https://cdn.intechopen.com/books/images_new/9759.jpg",slug:"vitamin-e-in-health-and-disease-interactions-diseases-and-health-aspects",publishedDate:"October 6th 2021",editedByType:"Edited by",bookSignature:"Pınar Erkekoglu and Júlia Scherer Santos",hash:"6c3ddcc13626110de289b57f2516ac8f",volumeInSeries:22,fullTitle:"Vitamin E in Health and Disease - Interactions, Diseases and Health Aspects",editors:[{id:"109978",title:"Prof.",name:"Pınar",middleName:null,surname:"Erkekoğlu",slug:"pinar-erkekoglu",fullName:"Pınar Erkekoğlu",profilePictureURL:"https://mts.intechopen.com/storage/users/109978/images/system/109978.jpg",institutionString:"Hacettepe University",institution:{name:"Hacettepe University",institutionURL:null,country:{name:"Turkey"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null}]},subseriesFiltersForPublishedBooks:[{group:"subseries",caption:"Proteomics",value:18,count:4},{group:"subseries",caption:"Metabolism",value:17,count:6},{group:"subseries",caption:"Cell and Molecular Biology",value:14,count:9},{group:"subseries",caption:"Chemical Biology",value:15,count:13}],publicationYearFilters:[{group:"publicationYear",caption:"2022",value:2022,count:8},{group:"publicationYear",caption:"2021",value:2021,count:7},{group:"publicationYear",caption:"2020",value:2020,count:12},{group:"publicationYear",caption:"2019",value:2019,count:3},{group:"publicationYear",caption:"2018",value:2018,count:2}],authors:{paginationCount:229,paginationItems:[{id:"318170",title:"Dr.",name:"Aneesa",middleName:null,surname:"Moolla",slug:"aneesa-moolla",fullName:"Aneesa Moolla",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/318170/images/system/318170.png",biography:"Dr. Aneesa Moolla has extensive experience in the diverse fields of health care having previously worked in dental private practice, at the Red Cross Flying Doctors association, and in healthcare corporate settings. 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:"419588",title:"Ph.D.",name:"Sergio",middleName:"Alexandre",surname:"Gehrke",slug:"sergio-gehrke",fullName:"Sergio Gehrke",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038WgMKQA0/Profile_Picture_2022-06-02T11:44:20.jpg",biography:"Dr. Sergio Alexandre Gehrke is a doctorate holder in two fields. The first is a Ph.D. in Cellular and Molecular Biology from the Pontificia Catholic University, Porto Alegre, Brazil, in 2010 and the other is an International Ph.D. in Bioengineering from the Universidad Miguel Hernandez, Elche/Alicante, Spain, obtained in 2020. In 2018, he completed a postdoctoral fellowship in Materials Engineering in the NUCLEMAT of the Pontificia Catholic University, Porto Alegre, Brazil. He is currently the Director of the Postgraduate Program in Implantology of the Bioface/UCAM/PgO (Montevideo, Uruguay), Director of the Cathedra of Biotechnology of the Catholic University of Murcia (Murcia, Spain), an Extraordinary Full Professor of the Catholic University of Murcia (Murcia, Spain) as well as the Director of the private center of research Biotecnos – Technology and Science (Montevideo, Uruguay). Applied biomaterials, cellular and molecular biology, and dental implants are among his research interests. He has published several original papers in renowned journals. In addition, he is also a Collaborating Professor in several Postgraduate programs at different universities all over the world.",institutionString:null,institution:{name:"Universidad Católica San Antonio de Murcia",country:{name:"Spain"}}},{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. 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