Important plants of the Solanaceae family and their uses [8].
\r\n\t
",isbn:"978-1-80356-273-5",printIsbn:"978-1-80356-272-8",pdfIsbn:"978-1-80356-274-2",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,isNomenclature:!1,hash:"e1d9662c334dd78ab35bfb57c3bf106e",bookSignature:"Dr. Fabio Arturo Iannotti",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11675.jpg",keywords:"Skeletal Muscle Diseases, Rare Skeletal Muscle Diseases, Basic Research, Molecular Mechanisms of Disease, Translational Research, Diagnostic Technologies, Functional Tests, Disease Models, Innovative Therapies, Drug Repositioning, Drug Discovery, Emerging Technologies",numberOfDownloads:24,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"February 11th 2022",dateEndSecondStepPublish:"April 19th 2022",dateEndThirdStepPublish:"June 18th 2022",dateEndFourthStepPublish:"September 6th 2022",dateEndFifthStepPublish:"November 5th 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"2 months",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dr. Fabio Arturo Iannotti received his Bachelor's Degree in Biotechnology Science at the University of Naples “Federico II” in 2006 with the highest degree. In 2010, he graduated with a Ph.D. in Neuroscience at the University of Naples “Federico II”. He published many papers on his areas of research in international peer-reviewed journals and for his pioneering studies has received awards from both national and international scientific societies.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"281317",title:"Dr.",name:"Fabio",middleName:"Arturo",surname:"Iannotti",slug:"fabio-iannotti",fullName:"Fabio Iannotti",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRdOdQAK/Profile_Picture_1644820016099",biography:"Currently researcher at the CNR-ICB Institute of Biomolecular Chemistry of Pozzuoli, Napoli (Italy), Fabio Arturo Iannotti has as major focus of his research activity the role of the endocannabinoid system and TRP in epilepsy and muscle development. Dr. Fabio Arturo Iannotti received his Bachelor Degree in Biotecnology Science (Medical curricula) at University of Naples \\'Federico II\\' in 2006 (with 110/110 cum laude). In 2010, Dr. Iannotti graduated with a PhD in Neuroscience at University of Naples \\'Federico II\\'. The focus of his thesis was on the role of voltage-gated potassium channels Kv7 during the neuronal excitoxicity as well as skeletal muscle cell differentiation. During the three years of the PhD program, Dr. Iannotti has been introduced to the field of ion channels, particularly voltage-gated ion channels; he has been instrumental in setting up RT-PCR and quantitative RT-PCR techniques in our lab, focusing onto research themes which would allow to combine both molecular and functional approaches in the study of ion channels during muscle cell differentiation. He has become familiar with most molecular biology (cloning, mutagenesis, PCR and RT-PCR, Southern and Northern blotting, gene silencing via RNAi, …) as well as with protein biochemistry techniques (protein extraction, immunoprecipitation, Western blotting, in-vitro translation, …) and morphological methods (confocal and conventional immunofluorescence). He is also familiar with imaging tools for intracellular ion concentration analysis, and has more recently gained considerable experience with electrophysiological techniques (specifically, patch-clamp). During this time (2009-2010), he also researched at the University of California-Davis assessing changes to the phosphorylation state of potassium channels in in vivo models of epilepsy. In 2011, he started his postdoc at the Institute of Biomolecular Chemistry (ICB)/ National Council of Research (CNR) and during this period he also visited the University of Reading (2012-2013), researching the potential involvement of TRP channels in epilepsy and muscle development. Since 2014, he was promoted to the position of research fellow at ICB. To date, Dr. Iannotti has published many papers on these areas of research in international peer reviewed journals, and has received awards from both national and international scientific societies for his work. 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According to [1], there is an increasing consumer preference for healthy foods, which has invited considerable demand for the use of anthocyanins as natural colorants, because of their natural pedigree and healthful properties. Anthocyanins are the most widely distributed group of water-soluble plant pigments in nature. They are mainly responsible for the mauve, red, blue, and purple colors in flowers, fruits, leaves, seeds and other organs in most of the flowering plants. The other important class of water-soluble pigments are betalains, which are present only in plants belonging to 13 families of Caryophyllales order [2-5]. An interesting phenomenon is the existence of mutual exclusiveness of anthocyanins and betalains in plant kingdom [3, 5-9]. Recent research demonstrated that simultaneous production of anthocyanins and betalains is possible in cell cultures and seedlings of anthocyanin producing plants by introduction and expression of genes encoding dihydroxyphenylalanine (L-DOPA) dioxygenases in combination with substrate precursor feeding [10]. However, the co-occurrence of both pigments in the same plant species have never been found in nature and the plants which produce anthocyanins never produce betalains and vice versa [6]. The commercial production of anthocyanin pigments is one of the fastest growing segments of the food colorant industry [2, 11]. The only industrial sources for anthocyanin pigments are from whole plant extracts [1], with the most common source being grape skins from the wine industry. According to [1], the demand of natural colorants continues to rise by 5-15% every year and this translated to the sales of anthocyanins isolated from grape skins in 2002, which was estimated to be US$200 million worldwide. The increase in demand for processed foods and high health products has caused the manufacturers to look for alternative sources of colorants with antioxidant properties. One source is the production of anthocyanins through the use of plant cell cultures [2, 12, 13].
Anthocyanins are synthesized via the flavonoid pathway, and they are known to contribute red, blue and purple color to colored grapes, wines and other products [14-18]. Anthocyanins can be used not only as food and beverage additives to obtain attractive natural coloration [19], but also for generating pharmaceutical and cosmetic products. Most researchers are optimistic about utilizing them as bioactive compounds with the consideration that they have the potential to improve human health [1]. Anthocyanins have been implicated in lowering the risk of cardiovascular disease and certain cancers. Dietary anthocyanins can be obtained by humans through the ingestion of fresh colored fruits processed into food and beverages. For instance, the consumption of red grapes and wine is considered vital for bioavailable anthocyanins [20, 21]. To date, most anthocyanin colorants are extracted from grape skins, black carrots, red cabbage, and sweet potato [11]. However, researchers are also exploring the idea of cultivating plant cell cultures for the production of natural colorants. Therefore, there is an interest in improving the quantity and quality of anthocyanins produced in grape cells, and this means that commercially viable systems must be developed to produce anthocyanins in grape cell cultures.
Production of anthocyanins by plant cell cultures is a feasible technology being pursued by industrial and academic interests. Several strategies are being used to enhance anthocyanin biosynthesis in plant cells. This involves a proper selection of the cell strain and optimization of media as well as culture conditions. It is crucial to note that anthocyanins obtained directly from fresh plant materials has limitations such as low metabolite yield, variability, and seasonal availability of raw materials, fresh material losses, inconsistent product quality, and pigment degradation caused by storage and extraction process [22]. Therefore, it is prudent to use
As colored molecules, anthocyanins play a key role in survival and evolution of flowering plants by attracting pollinators, frugivores and seed dispersers on one hand, and by repelling herbivores and parasites on the other [24-26]. Moreover, anthocyanins execute several important physiological functions in plant cells, and their biosynthesis is strongly induced by biotic and abiotic stress factors. These factors include, light, UV radiation, high or low temperatures, wounding, osmotic stress, nutrient imbalance, ozone exposure, herbivores, microbial and viral attacks. In [24, 27] the major roles of anthocyanins in photoprotection of chloroplasts from photoinhibitory damage have been discussed in details. The authors have also clarified the involvement of anthocyanins in protection from UV-B radiation, as well as how anthocyanins decrease oxidative stress by scavenging free radicals and modulating reactive oxygen signaling cascades. These cascades are responsible for triggering the expression of stress-responsive genes as well as the regulation of plant growth and development [24, 27].
Structurally anthocyanins are substituted glycosides and acylglycosides of 2-phenylbenzopyrilium salts (anthocyanidins). The basic structure of anthocyanidins consist of a chromane ring (C-6 – ring A and C-3 – ring C) bearing a second aromatic ring (C-6 – ring B) in position 2 (Figure 1) [2, 5, 28-30]. The various anthocyanidins differs in number and position of the hydroxyl and /or methyl ether groups attached on 3, 5, 6, 7, 3’, 4’ and/or 5’ positions. Despite the fact that 31 different monomeric anthocyanidins have been identified (including 3-deoxyanthocyanidins, pyranoanthocyanidins and sphagnorubins), 90% of the naturally occurring anthocyanins are based on only six structures (30% on cyanidin
Structures of common anthocyanidins
The color of anthocyanidins differs with the number of hydroxyl groups, attached on their molecules (especially those substituted in ring B). With the increase of attached hydroxyl groups, the visible color of entire molecule shift from orange to violet (Figure 2) [2, 5, 29, 30]. Glycosylation of anthocyanidins results to additional reddening of obtained anthocyanins, whereas the presence of aliphatic or aromatic acyl moieties causes no color change or slight blue shift and has significant effect on their stability and solubility [5]. Changes in pH can also cause reversible structural transformations in anthocyanins molecules, which has a dramatic effect on their color (Figure 3) [30-34].
Visible color range of common anthocyanidins
Structural transformations of anthocyanins in aqueous medium with different pH.
Most of the anthocyanins are
Anthocyanins based on cyanidin aglycone.
As a major flavonoid group, anthocyanins are products of phenylpropanoid metabolism of plant cells [28, 29, 38]. Anthocyanins in grapes are synthesized via flavonoid pathway. The biosynthetic pathway can be divided into two sections, the basic flavonoid upstream pathway, which includes early biosynthetic genes (EBGs), and the specific anthocyanin downstream branch, which includes late biosynthetic genes (LBGs) (Figure 5). Studies have shown that the basic flavonoid upstream pathway is restricted in many plants [39, 40, 41, 42] and that large gene families encodes the enzymes that act early in the flavonoid pathway, while the enzymes acting late in the pathway are encoded by single active gene [43]. The flavonoid pathway starts with phenylalanine, produced via shikimate pathway and transformed to 4-coumaroyl-CoA. The key enzyme, chalcone synthase (CHS) produce a naringenin chalcone by condense of one molecule 4-coumaroyl-CoA and three malonyl-CoA molecules (derived from citrate produced by The Krebs cycle) (Figure 5) [44]. In this case, the rings A and C are derived from the acetate pathway, whereas the ring B is derived from shikimate pathway [45]. Currently, there are three genes encoding CHS in grapes:
Flavonoids biosynthetic pathways and biosynthetic pathway of anthocyanins in grape: PAL - phenylalanine ammonia-lyase; C4H – cinnamate 4-hydroxylase; 4CL – 4-coumarate:CoA ligase; ACC – acetyl-CoA carboxylase; STS - stilbene synthase; CHS1, CHS2, and CHS3 - chalcone synthase 1, 2, and 3, respectively; PKR – polyketide reductase; AUS – aureusidin synthase; CHI1 and CHI2 - chalcone isomerase 1 and 2, respectively; FNS – flavone synthaes; FNR – flavonone 4-reductase; ANS – anthocyanidin synthase; GT – glucosyltransferases; AT – acyltransferases; MT – methyltransferases; F3’H – flavonoid 3’-hydroxylase; F3’5’H - flavonoid 3’5’-hydroxylase; F3H1 and F3H2 - flavanon 3β-hydroxylase 1 and 2, respectively; FLS – flavonol synthase; DFR – dihydroflavonol 4-reductase; LAR1 and LAR2 - leucoanthocyanidin reductase 1 and 2, respectively; ANR - anthocyanidin reductase;
After this reduction, anthocyanidin synthase (ANS) oxidize leucoanthocyanidins to their corresponding anthocyanidins. Anthocyanidins are inherently unstable under physiological conditions and were immediately glycosylated to anthocyanins by UDP-glucose: Anthocianidin: Flavonoid glucosyltransferase (UFGT) [48]. Anthocyanins, containing methylated anthocyanidins (peonidin
Once anthocyanins have been produced, they are transported and stored into the cell vacuole. Inside of vacuole, anthocyanins could be connected to specific proteins forming nomembrane intravacuolar bodies, known as anthocyanic vacuolar inclusions (AVI) (Figure 6) [50]. It has been confirmed that AVI plays a critical role in formation of color in flowering plants [50]. Recently, AVI from grape cell suspension were isolated and analyzed [51]. In contradiction with other plants, it was demonstrated that in grape cell suspension AVI consist of complex mix of tannins, anthocyanins (predominantly acylated derivates), proteins and other organic compounds, encased by lipid membrane [32, 51]. It was observed that a strong correlation between the prevalence of AVI structures in grapevine cell suspensions and the increase of their anthocyanin accumulation exist [51]. However, the enhancement of AVI prevalence does affect neither the number of available pigmented cells nor the overall growth rate of suspension cultures. Since AVI plays an important function in the storage and concentration of anthocyanins in cell vacuoles, their perspective role as enhancers of anthocyanin accumulation in grape cell suspensions have been proposed [51].
Anthocyanic vacuolar inclusions (AVI) in cell vacuoles of muscadine callus culture.
Grapes are rich sources of anthocyanins and bioavailable flavonoids. Grapevines are one of the world’s most grown economically important fruit crops. Currently, there are more than 10000 grape cultivars deposited in germplasm collection [52, 53]. Among them, the cultivars of
During their growth, grape berries follow a double sigmoid curve [68]. Veraison is the unique stage of berry development, representing the transition from growth stage to ripening. Once the grape berries enter to veraison, many physiological and biochemical changes occur. The grape cells completely redirect their metabolism to production of secondary metabolites, necessary to prepare berries for reaching the stage of physiological maturity. During this stage, the chlorophyll in berries has been completely lost and the biosynthesis of flavonoids, including anthocyanins is promoted [68]. Microarray analysis showed remarkable overexpression of genes involved in flavonoid biosynthesis and particularly in anthocyanin production during veraison and ripening in
Anthocyanin biosynthesis in grapes commences only when ripening of the berry begins (termed véraison) and normally continues throughout the ripening phase of growth. Anthocyanin biosynthesis pathway in grapes has been greatly investigated, including intracellular transportation and accumulation [18, 76-78]. Most of the structural genes have been isolated, cloned and characterized, and there is valuable information available on the mechanisms that regulate their expression within the plant cell [29, 44, 79]. Multiple regulatory genes under the complex regulation are responsible for the synthesis of anthocyanin at the transcriptional level [17, 18, 80]. The early biosynthetic genes (EBGs), which are upstream of the anthocyanin biosynthetic pathway, are regulated by several different families of genes called the Myb transcriptional factors, Myc transcriptional factors (encoding basic helix–loop-helix proteins, bHLH) and WD40-like proteins [40-42, 81]. On the other hand, late biosynthetic genes (LBGs), which are downstream leading to anthocyanin formation through glycosylation and subsequent modification (methylation and acylation) are under the specific control of several regulatory factors. Specific regulatory genes have been identified and characterized in
Recent studies in grapevine indicates that VvMYBA1 and VvMYBA2 transcription factors regulate UFGT gene, which plays a crucial role in the synthesis and accumulation of anthocyanins [56, 88-90], also identified another key R2R3-MYB protein that regulates proanthocyanidins (PA) synthesis in berry skin and seeds. But it is important to note that regulatory genes that control the expression of genes that encodes enzymes located upstream of UFGT have not been identified. It has been suggested that there is a contribution of at least two distinct regulatory complexes involved in the early and late steps of berry development [61]. Another study [86] also revealed that a MYB gene named
Muscadines (
Genetic engineering of plants has lots of benefit in the agricultural field [95-97]. It contributes to an efficient and cost-effective way to produce a wide array of novel, value-added plant and food products in an environmentally friendly manner. Most scientists including Butelli et al. [98] have highlighted the interest of producing crop plants and their products enriched with health-promoting natural compounds. These compounds include anthocyanins and flavonoids, which have become the targets for improving the nutritional value of foods. This requires an in depth knowledge of the molecular mechanisms underlying the biosynthetic pathways of secondary metabolites in plants as demonstrated by [98]. Even though there are some plants that contain high levels of anthocyanins such as blueberries, there are some species where the accumulation of secondary metabolites is not enough. This is why genetic engineering has been used as a strategy to modify flavonoid biosynthesis in order to enhance flower pigmentation in ornamentals and fruit plants [99-103]. Genetic engineering has become increasingly important worldwide because it provides significant improvements in the quantity, quality, and acceptability of the world\'s food supply and may be the best source for food security [5, 104]. Currently there is an increased production of plant-based products with an enhanced antioxidant capacity, which is facilitated by this technology [103]. An example of this approach is in tomato, which is also an important vegetable crop worldwide [98, 105, 106]. Several transgenic approaches have been used to enhance the accumulation of flavonoid levels in grape berries and tomato fruit by overexpressing either the structural or regulatory genes involved in the biosynthetic pathway [87, 107]. Most of these studies have been carried out
The final assembly of secondary metabolites in plants is determined by the coordinate transcriptional control of structural biosynthetic genes. Based on the information provided in [112], modulation of the rate of initiation of mRNA synthesis depends on the specific transcription factors, which interact with promoter regions of targeted genes. The regulatory genes that control the pattern and intensity of anthocyanin pigmentations through regulating the expression of several flavonoid-anthocyanin structural genes have been identified in many plants [16, 113-115]. There are two families of transcription factors i.e
In addition to the over-expression of the transcription factors, the suppression or negative regulators of flavonoid biosynthesis have been described [123]. For instance, high pigmented phenotypes (hp-2) in tomato were revealed when Bino et al. [123], mutated the DE-ETIOLATED1 gene (DET1). The fruits produced from these mutants are dark, and it is because of the elevated levels of flavonoids and carotenoids. The suppression of the regulatory gene
The regulatory gene families MYB and MYC control the structural genes within the grape anthocyanin biosynthetic pathway [87, 107]. But it is important to note that, the way in which the structural genes are regulated in grape berry skins appears to be different from the patterns observed in snapdragon, petunia, and maize [125-127]. There are two ways in which the pattern of gene expression in grape berry skins could be explained in relation to regulatory genes; 1.) early biosynthetic genes, which induces the expression of all of the structural genes except UFGT, and 2.) late biosynthetic genes that results in the induction of expression of all structural genes [1]. Alternatively, two types of regulatory genes may be present, one that controls expression of PAL, CHS, CHI, F3H, DFR and LDOX and another that induces UFGT gene expression [43]. This means that the regulatory gene that controls expression of PAL, CHS, CHI, F3H, DFR, and LDOX is expressed early in berry development. But it is crucial to note that many studies have identified UFGT, as the major control point to anthocyanin biosynthesis in grape berry skins, and this control is later in the pathway than has been observed in the studies of maize, petunia, and snapdragon anthocyanin biosynthesis.
Irrespective of the function of the regulatory genes, ectopic expression of the structural genes can also enhance the accumulation of anthocyanins. A study carried out by Muir et al. [108] determined that an ectopic expression of the Petunia
Silencing the structural or regulatory genes on the anthocyanin pathway in muscadine requires the following steps: 1) isolation of the endogenous structural or regulatory gene; 2) construction of the transformation cassettes using structural or regulatory-gene fragments as transgenes; 3) transformation of transgenic red cells via
The flow of genetic information dictates that “DNA is transcribed into RNA that is translated into a protein” (Fig 7A). Flavonoid biosynthetic proteins are produced using this concept. To shift the metabolic flux in muscadine grape cells, one can consider either 1) over-expressing the genes on the flavonoid pathway or 2) to knock out the production of the flavonoid proteins. Blocking the production of flavonoid proteins can be done by interfering with the flow of the genetic information. For example, to eliminate the production of the muscadine DFR protein (Fig 7B), we could interfere either at the mRNA transcription level (transcriptional gene silencing [TGS]) or at the post-transcriptional level (PTGS). The advancements made in genetic engineering have led to the possibility of knocking out the production of specific proteins in organisms by downregulating and/or silencing the genes encoding these proteins. Strategies developed to downregulate genes in plants include mutation-based reverse genetics [129], gene targeting [130], antisense RNA [131], cosuppression [95, 132], and RNA interference (RNAi) [133]. Genetic and biochemical evidences suggest that antisense-mediated gene silencing, co-suppression, and RNAi are all inputs into a common RNA silencing pathway triggered by the formation of a double-stranded RNA (dsRNA). This pathway, called PTGS, is characterized by accumulation of 21 to 25 nucleotides, small-interfering RNAs (siRNAs), sequence-specific degradation of target mRNA, and methylation of target gene sequences [134]. A typical example is demonstrated by Muir et al. [108], where they used RNAi to blocked specific metabolic conversions in the endogenous tomato flavonoid biosynthesis pathway by down-regulating the expression of specific structural flavonoid genes. In another study, Schijlen [135] also used RNAi technique to inhibit tomato
Schematic representation describing the flow of genetic information.
Grape cell suspension cultures have been extensively studied as an model for elucidation of anthocyanin biosynthesis pathway, for performing functional genetic studies, somatic embryo development and most importantly as an alternative source of natural colorants [1, 69, 140-142]. Cultivation of plant cells in controlled conditions offers advantages of continuous supply of high quality anthocyanin pigments. However, scientists have tried for over 40 years to produce anthocyanins in different cell systems, but until now, no commercially feasible anthocyanin producing system has been developed. Switching the production of natural pigments from the traditional approach (involving implementation of numerous agricultural activities) to modern industrial biotech factories is not an easy task. But it is obvious that for the successful realization of such biotechnological advances, research on both empirical and rational levels have to be performed [143]. To succeed in such challenges, we propose to follow a simple integrated approach based on consecutive conduction of various multidisciplinary experiments, optimization and monitoring procedures (Figure 8). As we already discussed some of the rational approaches for manipulation of anthocyanin biosynthesis on genetic level, in the next few pages, the basic principles of empirical studies are highlighted and the current progress on them has been reviewed.
Basic concepts for the development of biotechnological system for anthocyanins production based on grape cell suspension culture. The key steps in bioprocess engineering and optimization of anthocyanins production, involving application of both empirical and rational approaches are presented on each technological stage.
The first step for creation of biotechnology process for anthocyanins production is the development and selection of high producing cell line. To facilitate this process several important preliminary questions needs to be answered. The most important is the right choice of plant species (respectively, the appropriate cultivar) having the necessary anthocyanins profile in both quantity and quality aspects. Recently Lazar and Petolescu [144], generated cell suspension cultures of six grapevine varieties (Burgund Mare, Cabernet Sauvignon, Merlot, Oporto, Negru Tinctorial and Pinot Noir). They cultivated them in a laboratory bioreactor and demonstrated that the growth rates and biosynthetic potential for anthocyanins production were in strong dependence of cultivar used for culture initiation [144]. Currently, most of the research on grape cell suspensions have been performed with cell lines derived from two sources –
After the choice of targeted variety, the critical step is the selection of appropriate explants, which are used for callus initiation. The type and the age of explants, as well as the environmental conditions at which they are collected are critical factors for successful initiation of
Effects of the inoculum sizes (14 days old culture) on accumulated fresh weight (AFW) and packed cell volume (PCV) by
However, the non-homogenous growth of grape cell suspensions could be a serious issue for large-scale cultivation of these in
The optimal balance of nutrients in cultivation medium has been found to be an essential factor, determining the success on
Cell suspension of
As the grape cell suspensions exist as mixture of colored and colorless cells, the optimal cultivation medium should be developed by the way to provide a right balance between the growth rates of both cell populations [1, 142]. Finding the right nutrients balance is often a complicated task mainly because of the observation that the colorless population usually has better growth characteristics and the colored cells showed slow growth [1, 142, 150]. Dedaldechamp and Uhel [154] isolated a cell line from colorless cells of
Since the pH is an important factor for anthocyanin stability and activities of enzyme systems in plant cells, its value in the cultivation medium is critically important for the regulation of both pigments and biomass yields. Recently, the effect of pH in culture medium was investigated on callus cultures of three grapevine varieties (Coarnă neagră, Fetească neagră and Cadarcă) [164]. The authors observed that the largest amount of accumulated anthocyanins (13.5 mg/g FW) were registered in callus culture of Fetească neagră, cultivated on the medium with the pH=4.5. When cultivated on medium with pH=9.0, the anthocyanin production by the same culture was significantly decreased (up to 3.2 mg/g FW) [164]. Suzuki and colleagues [162], investigated the growth and production of anthocyanin in grape cell suspension from
Temperature has a strong effect on anthocyanin biosynthetic pathway, since some cold regulation genes are involved on it [165, 166]. Anthocyanin accumulation in berry skins of “Aki Queen” (
Light is an important controlling agent in anthocyanin biosynthesis [166]. Light has been found to induce the expression of genes, responsible for activation of the promoters of the flavonoid pathway genes (
Anthocyanin accumulation in callus culture of
Lazar and colleagues investigated the effect of light on anthocyanin accumulation by callus cultures from six grapevine varieties (Burgund Mare, Cabernet Sauvignon, Merlot, Oporto, Negru Tinctorial and Pinot Noir) [171]. They found that the light has a stimulating effect on anthocyanin production in all calli studied, but the amount of accumulated pigments was in strong correlation with the genotype of variety used for callus initiation [171]. In grape cell suspension of
Availability of growth regulators (auxins and cytokinins) in cultivation medium are essential for ensuring the growth and to determine the levels of produced secondary metabolite by
Application of different (biotic or abiotic) elicitors has been proved to be an effective strategy for enhancement of the production of secondary metabolites related to the plant defense system [175]. Anthocyanin biosynthetic pathway as a part of phenylpropanoid metabolism of plant cells could be significantly manipulated by application of different elicitors or feeding with specific precursors. Treatment of cell suspension culture of
The increased demand of natural colorants and nutraceuticals determines the needs for development of alternative technologies for supply of such additives. The anthocyanins, produced by grape cell suspensions, represent a very attractive class of natural compounds, which could find application in food industry (as colorants), pharmacy (as nutraceuticals and therapeutic compounds) and in cosmetics (as UV protectors, antioxidant and anti cancer compounds). Biotechnological production of grape anthocyanins presents significant economical benefits. Cormier and colleagues calculated that the cost of 1 kg anthocyanins, produced by two-stage cultivation process of grape cell suspension in bioreactor with working volume of 155,000 L can cost almost the half of the price of such amount of anthocyanins, produced by the extraction of grape skins ($ 931 per 1 kg of anthocyanins from grape cell suspension, compared to $ 2,083 per 1 kg of anthocyanins, produced by grape skin extraction) [142]. However, the specific requirements of the available grape cell suspensions significantly complicate the scale up of the cultivation process, which is the serious restriction for realization of such biotechnological process.
Anthocyanins have great potential for application in pharmaceutical products both as nutraceuticals and as therapeutic compounds. Frequent ingestion of anthocyanins could provide various health benefits including reduced risk of coronary heart diseases, anti-carcinogenic activity, antioxidant activity, reduced risk of stroke, anti-inflammatory effects etc. [13, 34, 184-186]. Biological activities of anthocyanin pigments have been already discussed in several excellent reviews [184, 187, 188]. Their pharmaceutical value has been additionally increased due to their high bioavailability. However, the administration and metabolism of anthocyanins
The world market of natural food colorants expands with the annual growth rate of 4-6% [142]. In USA 4 of the 26 colorants approved by the food administration, that are exempt from certification, are based on anthocyanin pigments [34]. In European Union, all anthocyanin-containing colorants are classified as natural colorants under the classification E163 [191]. Currently most of the worldwide anthocyanins supply comes from processing of grape pomace, which is a waste product from winemaking. But in European Union other plant sources such as red cabbage, elderberry, black currant, purple carrot, sweet potato, and red radish are also allowed [192]. Anthocyanins, produced by grape cell suspensions can be a promising alternative supply of natural colorants. It has already been demonstrated that the produced pigments by the grape cell suspensions undergo significant structural modifications. Grape cell suspensions accumulates higher levels of metabolically more evolved structures (methylated and acylated anthocyanins). Acylated anthocyanins are suitable for application in food products, mainly because of the improved color stability compared to non-acylated structures [72]. Moreover, the grape cell suspensions can also produce elevated levels of beneficial phenolic compounds such as flavonoids, stilbenes, phenolics, etc., which are capable of increasing the added value of the final additive. The overall metabolite profile of grape cells in combination with the lack of microbial and toxic contaminations will give the potential for development of new types of food additives if the entire cell suspension biomass are utilized.
The commercial interest of cosmetic companies to apply plant additives, derived by biotechnological cultivation of plant cells to their products has increased remarkably in the last few years [193]. The addition of plant cell derived extracts in cosmetic products has been considered as a powerful approach used to increase their health benefits. Several plant extracts have been added to various cosmetic products as moisturizers, antioxidants, whitening agents, colorants, sunscreens, preservatives etc. [193]. With the advancement of plant cell biotechnology, more and more cosmetic companies have been attracted for application of additives, based on plant cell suspensions. Recently the application of so-called plant “steam” cells attracts industry’s attention [193]. In the last few years, the French company “Sederma” launched the product “Resistem™” based on application of
The approaches described in this chapter can be effective in improving novel anthocyanin-derived metabolites in grape cell suspensions. Continuous study and exploitation of the knowledge of grape cell lines and their control mechanisms will open up new possibilities for metabolic engineering of the anthocyanin biosynthesis pathway. In parallel, the recent achievements in bioengineering with plant cell suspensions and the improvements of the existed bioreactor designs discovers new prospectives for commercial realization of anthocyanin producing technology based on cultivation of grape cells. This is a research area that is growing and gaining interest in the analysis of plant-based health-related compounds. Therefore, the full impact of metabolomics on muscadine research is yet to be experienced. But this chapter serves as a starting point for scientists who are interested in cell cultures from muscadine grapes.
Plants have been used for the treatment of diseases for centuries ago. Ancient manuscripts of different civilizations show evidence of using herbs as medicine. Now, this system is used in Unani, Ayurveda, and Siddha medicines also. According to WHO, 21,000 plant species have the potential to use as medicinal plants [1]. The plant is used as a whole or parts as medicine. The plant contains various chemicals called phytochemicals or phytonutrients, which are primary metabolites or secondary metabolites. These metabolites protect the plants from the attack of microbes, such as bacteria, fungi, and viruses. These chemicals are rich in fruits, vegetables, grains, and other plants. The intake of these plants decreases the risk of developing cancer, diabetes, and heart diseases. These chemicals may act as antioxidants or nutrient protectors [2].
Cancer, the abnormal growth of cells that can invade the nearby cells and even spread to other organs called metastasis, which can occur in any part of the body results due to various causes and factor is one of the most dreadful conditions in the world. Common cancer reported are breast, lung, colon, rectum, etc. Among those mentioned types high-mortality rate reported was because of lung cancer. The number of people affected is male [3]. The common treatment methods are radiation therapy, chemotherapy, and surgery. These treatments are quite expensive and also possess many side effects. These side effects can be reduced by using plant or plant-based drugs. Scientists prove that several plants and their components have the potential to fight against cancer and mechanism of action include cessation of the cell cycle, regulation of the transcription process, induction of autophagy, downregulation of proteins in biochemical pathways, and rupturing of the membrane. Also, it was reported that some chemicals are effective against more than one type of cancer [4].
There are different classes of phytochemicals derived anticancer drugs available on market. The drugs were used as tinctures, tea, powders, decoctions, etc.
One among the family which possesses anticancer properties is the Solanaceae family. It is an angiosperm with 102 genera and more than 3000–4000 species of plants. Glycoalkaloids from Solanum species, such as solanine, solasonine, and solanidine, isolated from
Solanaceae family belongs to the order Solanales of angiosperms. Generally, they are called as potato family. This group is also called as nightshade family because of the poisonous alkaloids present in some members of the family. Most of the plants are economically important because of their food, ornamental or medicinal values. Some of the plants in this family are potato, tomato, all peppers, eggplant, etc. It also contains deadly toxic plants. These family members are found throughout the world but are widely distributed in the tropical regions of South America. Members of this family show different morphological and ecological characteristics [6]. Some of the medicinal plants are
Genus name | Plant name | Uses |
---|---|---|
Solanum | Used for dysentery, stomach complaints, antitumorigenic, and antioxidant. | |
used for the treatment of rheumatic pains, eye diseases, heart pain | ||
lower blood cholesterol level, used in the treatment of internal bleeding, piles, and toothache | ||
Used against cough, sore throat, etc. | ||
To prevent prostate cancer, breast cancer, and skin diseases | ||
Atropa | Used as diuretics | |
Used in traditional medicine | ||
Capsicum | Lowering blood pressure and Cholesterol. | |
Treatment of cancer | ||
Datura | antispasmodic and antiasthmatic diabetes, cancer, and viral infections | |
Withania | Used as a tranquilizer, Cancer treatment | |
Is used to treat nervous prostration, sleeplessness, infertility, multiple sclerosis, etc. | ||
Hyoscyamus | Cerebral and spinal sedative Used as anesthetics | |
Nicotiana | antispasmodics, diuretics, antioxidant activity along with other pharmacological effects | |
Physalis | Used as a hallucinogen, used as a remedy for abscesses, coughs, fevers, and sore throats. | |
Scopolia | Used as antispasmodic |
Important plants of the Solanaceae family and their uses [8].
Afroz et al. reported, “Bioactive secondary metabolites reported from the members of the Solanaceae include AMPs, alkaloids, flavonoids, glycosides, lactones, lignans, steroids, simple phenols, sugars, and terpenoids” [7].
Phytochemicals or phytonutrients are produced by plants. These chemicals are the basic principle for the nutritive as well as pharmacological action of plants. These chemicals also provide a defense mechanism to the plants against plant pathogens. The main pathogens belong to the class of bacteria, fungi, protozoa, and viruses [9]. The common vegetables and fruits, such as broccoli, berries, carrot, tomato, garlic, seeds, and onion, contain these chemicals in large quantities which increase the immune capacity of the body.
There are two types of metabolites in all living organisms including plants. They are primary metabolites, such as sugar, amino acids, nucleotides, and lipids, which are essential for their functioning. Plants also produce certain molecules which are not used directly in their life process such chemicals are called secondary metabolites synthesized by biochemical pathways, such as alkaloids, flavonoids, glycosides, phenolics, and terpenoids discovered with various properties, such as induction of flowering, protection from pathogens, protection from the external environment, attractant or repellant for pollination, antimicrobial, antioxidant, anti-inflammatory, and antitumor activity. These chemicals and their product are used in herbal medicine and modern medicines [10, 11, 12].
The phytochemicals are classified on the basis of their biosynthetic origin, structure, and solubility properties. The different types of phytochemicals are alkaloids, glycosides, flavonoids, saponins, terpenes, steroids, etc. [13]. The Solanaceae family is rich in phytochemicals showing their effectiveness as medicinal plants (Table 2). The review studies show that most of these common plants in the selective family show anticancer properties (Figure 2) which can enlighten the treatment of cancer in the near future.
Genus name | Plant name | Phytochemical | Class |
---|---|---|---|
Solanum | Solasonine, Solamargine, Solanigroside P Solamargine | Steroidal glycoalkaloids | |
Anguivioside A,B,C | Saponins | ||
Solamargine, Methylprotodioscin Indioside D | Steroidal glycoalkaloids | ||
Lycopene. Diosgenin | Carotenoid Saponins | ||
Atropine | Alkaloids | ||
Atropine, Apoatropine Scopolamine Kaempferol 3,7-diglycosides | Alkaloids Alkaloids Alkaloids Flavonoids | ||
Capsicum | Caffeic acid, capsinoids Canusesnol kaempferol-3-O-glucoside | Phenolics sesquiterpenoids flavonols | |
Capsaicin | capsaicinoids | ||
Datura | Apoatropine Tropine tropate Withanolides, | Nitrogen-containing polyhydroxylated heterocyclic compounds steroidal lactones | |
Withania | Withanolides, Withaferin A, Physagulin D, Withanoside IV | Steroids steroidal lactone Glycoside Glycoside | |
Withanolides | Steroidal lactone | ||
Hyoscyamus | Hyoscyamine, Apo atropine, hyoscine, skimmianine, scopolamine, belladonines | Alkaloids | |
Nicotiana | Nicotine | Alkaloid |
Plants of Solanaceae family and their phytochemicals against cancer.
The use of phytochemicals in pharmaceutical and agrochemical industries is an ongoing process that requires continuous and elaborate study.
Alkaloids are cyclic nitrogenous secondary metabolites seen in many plants. They are synthesized during the biochemical synthesis of proteins and nucleic acid. These alkaloids have wide applications as drugs, narcotics, or poisons. Jerzykiewicz et al. reported, “Chenopodiaceae, Lauraceae, Magnoliaceae, Berberidaceae, Menispermaceae, Ranunculaceae, Papaveraceae, Fumariaceae, Papilionaceae, Rutaceae, Apocynaceae, Loganiaceae, Rubiaceae, Boraginaceae, Convolvulaceae, Solanaceae, and Campanulaceae are some of the families that rich in alkaloids which protect the plant from insects, pest and also give disease resistant capacity to plants” [18]. This property of alkaloids in Solanaceae can be utilized for anticancer medicine production.
Alkaloids are classified into different categories based the on nature of the precursor molecule for its biosynthesis, chemical structure, biological effect, and heterocyclic or non-heterocyclic types [19]. The biological effects of alkaloids include hallucinogens, antimalarial, tranquilizer, anticancer, CNS stimulant, insecticidal, antiviral, antihypertension, antimicrobial, antirheumatics, anti-inflammatory, antioxidant, and diuretics. The alkaloids are used in the drug industry because they are the precursor for medicines for cardiovascular disease, menopause, etc.
Alkaloid | Type of Alkaloid | Source | Biological Action |
---|---|---|---|
Atropine | Tropane | Para sympatholytic, Anticholinergic. | |
Hyoscyamine | Tropane | ||
Scopolamine | Tropane | Antidepressant and Antinausea | |
Daturametelindoles A-D (1–4) | Indole Alkaloids | Anticancer Effect | |
Anabasine | Pyridine Alkaloids | Insecticide | |
Nicotine | Pyridine Alkaloids | Insecticidal | |
Chaconine | Glycoalkaloid | Fungicidal | |
Solanidane; Solanidine | Steroidal Alkaloid | Anticancer | |
Solamargine | Glycoalkaloid | anticancer | |
Solamarine | Glycoalkaloid | Antibacterial | |
Solanine | Saponins | Antifungal | |
Solanopubamine | Steroidal alkaloid | Anticancer | |
Solasodine | glycoalkaloid | Antibacterial | |
Solasonine | glycoalkaloid | substrate for the production of important steroids | |
Tomatidine | steroid glycosides | Anticancerous |
The review studies show that tomatidine, solanopubamine, solamargine, solanidane; solanidine, daturametelindoles A-D (1–4) chemicals show anticancer activity in in vitro conditions by activation of caspase-3 and regulation of cell cycle to induce apoptosis. The detailed mechanism of action of these chemicals and their clinical trials will be an asset for developments in cancer medicine.
Flavonoids have several potential effects in plants system such as attracting pollination, seed germination, aromatic flavors in defense mechanisms, stress tolerance, UV photoprotection, inducing root nodulation, and controlling transport of plant hormones [25, 26, 27]. Panche et al. reviewed that flavonoids have pharmaceutical, medicine, and cosmetic applications.
The different polyphenolic and glycosidic compounds had been reported from various members of the Solanaceae family (Table 4). Scopoletin (7-hydroxy-6-methoxycoumarin), a coumarin, was isolated from
Sl | Flavonoids | Plant Source | Biological Action |
---|---|---|---|
1 | Luteolin | Anticancer | |
2 | Apigenin | Antioxidant,antibacterial, cytotoxic | |
3 | Kaempferol | Anticancer | |
4 | Quercetin | Anticancer | |
5 | Tangeretin | Antioxidant, anti-inflamotory,antitumour | |
6 | Mycicetin | Antispasmodic effect | |
7 | Scopoletin | Hepatoprotective activity | |
8 | Anthocyanidins | ||
9 | Anthocyanin | Inhibition of cell proliferation |
The flavonoids, such as apigenin, kaempferol, quercetin, and anthocyanin are some of the chemicals isolated from the respective family that possesses antiproliferation effects against cancer cell lines [30].
More than 100 plant families, a few starfishes, and sea cucumber reported the presence of saponins. Dicot families, such as Leguminosae, Araliaceae, and Caryophyllaceae, are sources of triterpenoid saponins. Steroidal saponins are found in families, such as Agavaceae, Alliaceae, Asparagaceae, Dioscoreaceae, Liliaceae, Amaryllidaceae, Bromeliaceae, Palmae, and Scrophulariaceae. Solanaceae families contain steroidal glycoalkaloids [31]. Some of the biological activities of saponins are anti- cancer activity, reducing cholesterol levels, decreasing blood glucose, anti-inflammatory potentials, antibacterial, antifungal and antiviral activity [32, 33].
Figueiredo et al. isolated steroidal saponins from the roots of
Terpenes are used in herbal medicines because of their biological activities. Some of the activities include antiplasmodial, especially antimalarial, anticancer, antidiabetic, anti-inflammatory, antioxidant, etc. Curcumin is one of the terpenes used in folk medicine Terpenes are used as flavors and fragrances in food and cosmetics [36].
Diterpene phytol was isolated from Solanum schimperianum, and Betulinic acid was isolated from Solanum buddleifolium. 3β-Hydroxysolavetivone from the root of S. abutiloides showed antifungal activities. Solavetivone and Lubimin also showed antifungal activities isolated from
The common conventional treatment of cancer causes side effects and drug resistance in patients, so many plant species were attempted as anticancer drugs. α-chaconine a derivative of solanidine shows an antimetastatic effect individually also in combination with gallic acid by caspase-dependent apoptosis Reddivari et al. [37]. Solanine a glycoalkaloid present in
Solamargine glycoalkaloid present in
Saponins from
The above-mentioned plants exhibit anticancer mechanisms by cell cycle arrest: The cell cycle contains several proteins at the checkpoint. Cancer cells overcome this checkpoint leads to the multiplication of cells. So inducing cell cycle arrest can be an alternative method in the treatment of cancer. Various researches show that this is possible by phytochemicals. The phytochemicals, such as solanine, solanidine, solamargine, and α-chaconine, result in cell cycle arrest at the S phase of the cell cycle and thereby induce apoptosis based on various concentrations. [52] (Figure 3).
Mechanism of cell cycle inhibition by phytochemicals of Solanaceae [
Studies prove that these chemicals have anticancer properties against different types of cancer, such as breast cancer, colon cancer, cervical cancer, and liver cancer.
Another mechanism involved in anticancer therapy is the regulation of transcription by inhibiting oncogenic transcription factors. Withaferin isolated from Withania sominifera shows antitumor activity by regulating transcription factors [5].
Physapubescin B and physapubenolide isolated from Physalis pubescens exhibit anticancer mechanisms by autophagy (Figure 4) and apoptosis in colorectal cancer cell lines [53].
Anticancer mechanism of physapubenolide [
Other mechanisms include the suppression of metabolic enzymes. Some plant molecules can cause apoptosis by breaking the mitochondrial membrane. Defensin isolated from Nicotiana alata induces necrotic-like cell death in a number of tumor cells [5]. The summary of the anticancer potential of selected plants that are economically useful to humans of the Solanaceae family with the mechanism of their action against different cancer cell lines which is reviewed in this literature from previous works by eminent workers is given in the table (Table 5).
Plant Source | Phytochemical | Model | Mechanism of Action |
---|---|---|---|
Solanine | Human pancreatic cancer cell lines, human melanoma cell lines, human prostate cancer cells | Apoptosis | |
Solanidine | Human lung adenocarcinoma cell line (A549) A549 CAM xenograft BALB/c mouse model | Inhibition of DNA synthesis | |
Tomatidine | Human fibrosarcoma cells, human lung adenocarcinoma cell A549 | Suppression of cell invasion by inhibition of ERK and Akt signaling pathways | |
Tomatine | Prostate cancer in mice | Induction of apoptosis mediated by P13K/Akt pro-signaling pathway | |
Solamargine | Human liver cancer cell lines, i.e., HepG2 and Huh-7 cells Human neuroblastoma cell line (SH-SY5Y) | By arresting the cell cycle at the G2/M phase | |
Solasodine | Human colorectal cancer cells | Suppression of the AKT/glycogen synthase kinase-3β/β-catenin pathway | |
Capsaicin | Human cell lines of different origins | Apoptosis, cell-cycle arrest, transcription factor regulation | |
Withanolides | Human colorectal carcinoma | Inhibit tumor cell proliferation | |
Nicotine | Human airway epithelial cells. | By regulation of tumor necrosis factor | |
Anthocyanins | Different cancer cell models | Inhibition of cell multiplication and apoptosis | |
Degalactotigonin | Human Pancreatic cancer cell lines | Induces apoptosis and cell cycle arrest by inhibiting the signaling pathway | |
Saponins | Human larynx cancer cell lines | Initiation of apoptosis | |
Withaferin | Chicken myeloid cell lines (HD11-C3-GFP1 Myb reporter cell line) Human Myeloid leukemia cell line (HL60) Quail Japanese fibrosarcoma (QT6) Mouse preadipocyte cell line (3T3-L1) | By regulating transcription factors | |
Physapubescin B and physapubenolide | Colorectal cancer cell lines. | Autophagy and apoptosis |
The effective study of the mechanism of phytochemicals in the Solanaceae family will open a new approach to the treatment of cancer.
Many of these are commercially interesting because of their use as flavors and fragrances in foods and cosmetics.
Many of these are commercially interesting because of their use as flavors and fragrances in foods and cosmetics.
Cancer is one of the major public health problems across the world. The pandemic condition of the current world results in the delay of diagnosis and treatment that may lead to increased complications in the treatment of cancer. Phytochemicals from Solanaceae exhibit anticancer activity against various type of cancer.
These compounds proved their efficiency in the inhibition of cancer cell line proliferation by cell cycle arrest, regulation of transcription factors, blocking the signal pathways, initiation of apoptosis, and suppression of metastasis. Most of the compound shows positive results with a combination of other phytochemicals in cancer treatment. The effective study of these biomolecules as anticancer targets can lead to clinical trials and in the future, it opens an effective area for the treatment of cancer and prevention.
The author acknowledges and thanks to Karpagam Academy of Higher Education, Coimbatore providing the internet facility to complete this review process.
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Muedi",authors:[{id:"225304",title:"Dr.",name:"Vhahangwele",middleName:null,surname:"Masindi",slug:"vhahangwele-masindi",fullName:"Vhahangwele Masindi"},{id:"241403",title:"M.Sc.",name:"Khathutshelo",middleName:"Lilith",surname:"Muedi",slug:"khathutshelo-muedi",fullName:"Khathutshelo Muedi"}]},{id:"36171",doi:"10.5772/36942",title:"Research of Calcium Phosphates Using Fourier Transform Infrared Spectroscopy",slug:"research-of-calcium-phosphates-using-fourier-transformation-infrared-spectroscopy",totalDownloads:9225,totalCrossrefCites:130,totalDimensionsCites:375,abstract:null,book:{id:"1591",slug:"infrared-spectroscopy-materials-science-engineering-and-technology",title:"Infrared Spectroscopy",fullTitle:"Infrared Spectroscopy - Materials Science, Engineering and Technology"},signatures:"Liga Berzina-Cimdina and Natalija Borodajenko",authors:[{id:"110522",title:"Prof.",name:"Liga",middleName:null,surname:"Berzina-Cimdina",slug:"liga-berzina-cimdina",fullName:"Liga Berzina-Cimdina"},{id:"112181",title:"MSc.",name:"Natalija",middleName:null,surname:"Borodajenko",slug:"natalija-borodajenko",fullName:"Natalija Borodajenko"}]},{id:"41411",doi:"10.5772/53659",title:"Textile Dyes: Dyeing Process and Environmental Impact",slug:"textile-dyes-dyeing-process-and-environmental-impact",totalDownloads:20608,totalCrossrefCites:97,totalDimensionsCites:305,abstract:null,book:{id:"3137",slug:"eco-friendly-textile-dyeing-and-finishing",title:"Eco-Friendly Textile Dyeing and Finishing",fullTitle:"Eco-Friendly Textile Dyeing and Finishing"},signatures:"Farah Maria Drumond Chequer, Gisele Augusto Rodrigues de Oliveira, Elisa Raquel Anastácio Ferraz, Juliano Carvalho Cardoso, Maria Valnice Boldrin Zanoni and Danielle Palma de Oliveira",authors:[{id:"49040",title:"Prof.",name:"Danielle",middleName:null,surname:"Palma De Oliveira",slug:"danielle-palma-de-oliveira",fullName:"Danielle Palma De Oliveira"},{id:"149074",title:"Prof.",name:"Maria Valnice",middleName:null,surname:"Zanoni",slug:"maria-valnice-zanoni",fullName:"Maria Valnice Zanoni"},{id:"153502",title:"Ph.D.",name:"Farah",middleName:null,surname:"Chequer",slug:"farah-chequer",fullName:"Farah Chequer"},{id:"153504",title:"MSc.",name:"Gisele",middleName:null,surname:"Oliveira",slug:"gisele-oliveira",fullName:"Gisele Oliveira"},{id:"163377",title:"Dr.",name:"Juliano",middleName:null,surname:"Cardoso",slug:"juliano-cardoso",fullName:"Juliano Cardoso"},{id:"163393",title:"Dr.",name:"Elisa",middleName:null,surname:"Ferraz",slug:"elisa-ferraz",fullName:"Elisa Ferraz"}]},{id:"17237",doi:"10.5772/24553",title:"Hydrogels: Methods of Preparation, Characterisation and Applications",slug:"hydrogels-methods-of-preparation-characterisation-and-applications",totalDownloads:65847,totalCrossrefCites:86,totalDimensionsCites:277,abstract:null,book:{id:"248",slug:"progress-in-molecular-and-environmental-bioengineering-from-analysis-and-modeling-to-technology-applications",title:"Progress in Molecular and Environmental Bioengineering",fullTitle:"Progress in Molecular and Environmental Bioengineering - From Analysis and Modeling to Technology Applications"},signatures:"Syed K. H. Gulrez, Saphwan Al-Assaf and Glyn O Phillips",authors:[{id:"58120",title:"Prof.",name:"Saphwan",middleName:null,surname:"Al-Assaf",slug:"saphwan-al-assaf",fullName:"Saphwan Al-Assaf"}]}],mostDownloadedChaptersLast30Days:[{id:"35255",title:"Mechanical Transmissions Parameter Modelling",slug:"mechanical-transmissions-parameter-modelling",totalDownloads:7279,totalCrossrefCites:1,totalDimensionsCites:2,abstract:null,book:{id:"1982",slug:"mechanical-engineering",title:"Mechanical Engineering",fullTitle:"Mechanical Engineering"},signatures:"Isad Saric, Nedzad Repcic and Adil Muminovic",authors:[{id:"101313",title:"Prof.",name:"Isad",middleName:null,surname:"Saric",slug:"isad-saric",fullName:"Isad Saric"}]},{id:"68505",title:"Research Design and Methodology",slug:"research-design-and-methodology",totalDownloads:24813,totalCrossrefCites:7,totalDimensionsCites:16,abstract:"There are a number of approaches used in this research method design. The purpose of this chapter is to design the methodology of the research approach through mixed types of research techniques. The research approach also supports the researcher on how to come across the research result findings. In this chapter, the general design of the research and the methods used for data collection are explained in detail. It includes three main parts. The first part gives a highlight about the dissertation design. The second part discusses about qualitative and quantitative data collection methods. The last part illustrates the general research framework. The purpose of this section is to indicate how the research was conducted throughout the study periods.",book:{id:"8511",slug:"cyberspace",title:"Cyberspace",fullTitle:"Cyberspace"},signatures:"Kassu Jilcha Sileyew",authors:[{id:"292841",title:"Ph.D.",name:"Kassu",middleName:null,surname:"Jilcha Sileyew",slug:"kassu-jilcha-sileyew",fullName:"Kassu Jilcha Sileyew"}]},{id:"67558",title:"Polymerase Chain Reaction (PCR): Principle and Applications",slug:"polymerase-chain-reaction-pcr-principle-and-applications",totalDownloads:10511,totalCrossrefCites:6,totalDimensionsCites:15,abstract:"The characterization of the diversity of species living within ecosystems is of major scientific interest to understand the functioning of these ecosystems. It is also becoming a societal issue since it is necessary to implement the conservation or even the restoration of biodiversity. Historically, species have been described and characterized on the basis of morphological criteria, which are closely linked by environmental conditions or which find their limits especially in groups where they are difficult to access, as is the case for many species of microorganisms. The need to understand the molecular mechanisms in species has made the PCR an indispensable tool for understanding the functioning of these biological systems. A number of markers are now available to detect nuclear DNA polymorphisms. In genetic diversity studies, the most frequently used markers are microsatellites. The study of biological complexity is a new frontier that requires high-throughput molecular technology, high speed computer memory, new approaches to data analysis, and the integration of interdisciplinary skills.",book:{id:"7728",slug:"synthetic-biology-new-interdisciplinary-science",title:"Synthetic Biology",fullTitle:"Synthetic Biology - New Interdisciplinary Science"},signatures:"Karim Kadri",authors:[{id:"290766",title:"Dr.",name:"Kadri",middleName:null,surname:"Karim",slug:"kadri-karim",fullName:"Kadri Karim"}]},{id:"62059",title:"Types of HVAC Systems",slug:"types-of-hvac-systems",totalDownloads:12245,totalCrossrefCites:8,totalDimensionsCites:14,abstract:"HVAC systems are milestones of building mechanical systems that provide thermal comfort for occupants accompanied with indoor air quality. HVAC systems can be classified into central and local systems according to multiple zones, location, and distribution. Primary HVAC equipment includes heating equipment, ventilation equipment, and cooling or air-conditioning equipment. Central HVAC systems locate away from buildings in a central equipment room and deliver the conditioned air by a delivery ductwork system. Central HVAC systems contain all-air, air-water, all-water systems. Two systems should be considered as central such as heating and cooling panels and water-source heat pumps. Local HVAC systems can be located inside a conditioned zone or adjacent to it and no requirement for ductwork. Local systems include local heating, local air-conditioning, local ventilation, and split systems.",book:{id:"6807",slug:"hvac-system",title:"HVAC System",fullTitle:"HVAC System"},signatures:"Shaimaa Seyam",authors:[{id:"247650",title:"M.Sc.",name:"Shaimaa",middleName:null,surname:"Seyam",slug:"shaimaa-seyam",fullName:"Shaimaa Seyam"},{id:"257733",title:"MSc.",name:"Shaimaa",middleName:null,surname:"Seyam",slug:"shaimaa-seyam",fullName:"Shaimaa Seyam"},{id:"395618",title:"Dr.",name:"Shaimaa",middleName:null,surname:"Seyam",slug:"shaimaa-seyam",fullName:"Shaimaa Seyam"}]},{id:"70315",title:"Some Basic and Key Issues of Switched-Reluctance Machine Systems",slug:"some-basic-and-key-issues-of-switched-reluctance-machine-systems",totalDownloads:1238,totalCrossrefCites:0,totalDimensionsCites:1,abstract:"Although switched-reluctance machine (SRM) possesses many structural advantages and application potential, it is rather difficult to successfully control with high performance being comparable to other machines. Many critical affairs must be properly treated to obtain the improved operating characteristics. This chapter presents the basic and key technologies of switched-reluctance machine in motor and generator operations. The contents in this chapter include: (1) structures and governing equations of SRM; (2) some commonly used SRM converters; (3) estimation of key parameters and performance evaluation of SRM drive; (4) commutation scheme, current control scheme, and speed control scheme of SRM drive; (5) some commonly used front-end converters and their operation controls for SRM drive; (6) reversible and regenerative braking operation controls for SRM drive; (7) some tuning issues for SRM drive; (8) operation control and some tuning issues of switched-reluctance generators; and (9) experimental application exploration for SRM systems—(a) wind generator and microgrid and (b) EV SRM drive.",book:{id:"8899",slug:"modelling-and-control-of-switched-reluctance-machines",title:"Modelling and Control of Switched Reluctance Machines",fullTitle:"Modelling and Control of Switched Reluctance Machines"},signatures:"Chang-Ming Liaw, Min-Ze Lu, Ping-Hong Jhou and Kuan-Yu Chou",authors:[{id:"37616",title:"Prof.",name:"Chang-Ming",middleName:null,surname:"Liaw",slug:"chang-ming-liaw",fullName:"Chang-Ming Liaw"},{id:"306461",title:"Mr.",name:"Min-Ze",middleName:null,surname:"Lu",slug:"min-ze-lu",fullName:"Min-Ze Lu"},{id:"306463",title:"Mr.",name:"Ping-Hong",middleName:null,surname:"Jhou",slug:"ping-hong-jhou",fullName:"Ping-Hong Jhou"},{id:"306464",title:"Mr.",name:"Kuan-Yu",middleName:null,surname:"Chou",slug:"kuan-yu-chou",fullName:"Kuan-Yu Chou"}]}],onlineFirstChaptersFilter:{topicId:"1",limit:6,offset:0},onlineFirstChaptersCollection:[{id:"82418",title:"Bayesian Networks for Decision Support in Emergency Response: A Model for Missing Person Investigations",slug:"bayesian-networks-for-decision-support-in-emergency-response-a-model-for-missing-person-investigatio",totalDownloads:0,totalDimensionsCites:0,doi:"10.5772/intechopen.105047",abstract:"The successful operation of Emergency services (Police, Fire, Medical Emergency) relies heavily upon Information Systems and particularly Decision Support Systems. Missing person cases consume resources from the already overstretched resources of Police Forces. Such cases predominantly come from at-risk groups such as children in care, people suffering from depression, or elderly people suffering from dementia. This chapter reviews current practices used for missing person cases and describes a decision support model based on Bayesian networks.",book:{id:"11068",title:"Contemporary Issues in Information Systems - a Global Perspective",coverURL:"https://cdn.intechopen.com/books/images_new/11068.jpg"},signatures:"Denis Reilly"},{id:"82413",title:"Utilization of Biopolymers in Water Based Drilling Muds",slug:"utilization-of-biopolymers-in-water-based-drilling-muds",totalDownloads:0,totalDimensionsCites:null,doi:"10.5772/intechopen.105516",abstract:"With the increase in energy demand, deeper wells drilling is one of the solutions to fulfill the energy demand, which demands specialized drilling mud formulation. These muds are composed of thermally stable materials that can sustain in high-temperature conditions. Biopolymers are widely used out of various mud additives for improving the rheology and filtration characteristics of mud. Owing to the high temperature and poor thermal stability of such additives, these additives lose their primary functions, resulting in the nonproductive time and irreversible problems. The book chapter highlights the uses of water-based mud, its limitations, and the degradation of biopolymers. Various additives’ significance and susceptibility in harsh borehole conditions have been discussed. The existing additives used for the rheological and filtration characteristics improvements and their shortcomings are presented. Furthermore, the field applications of native and modified polymeric-based mud formulations have been further examined and presented.",book:{id:"11929",title:"Drilling Engineering and Technology - Recent Advances, New Perspectives and Applications",coverURL:"https://cdn.intechopen.com/books/images_new/11929.jpg"},signatures:"Imtiaz Ali, Maqsood Ahmad, Aftab Hussain Arain, Vahid Atashbari and Asif Zamir"},{id:"82123",title:"Microwave-Assisted Pyrolysis Process: From a Laboratory Scale to an Industrial Plant",slug:"microwave-assisted-pyrolysis-process-from-a-laboratory-scale-to-an-industrial-plant",totalDownloads:0,totalDimensionsCites:null,doi:"10.5772/intechopen.104925",abstract:"One of the great challenges for the European Union (EU) is the “Circular Economy Package,” and to achieve this goal, materials at the end of their life cycle must be recycled using a sustainable process. In this way, as a thermochemical treatment, pyrolysis represents a significant opportunity so long it leads to the recovery of both energy and chemical content of mixed, contaminated, or deteriorated plastics. An excellent history of an academic-industrial adventure started in 2008 at the Department of Chemistry of the University of Florence demonstrates the possibility of employing microwaves to recycle plastics to preserve their energy and chemical content. After that, Techwave started industrialization of the process in 2019, realizing a small-scale prototype followed by a full-scale pilot plant using different plastic materials (e.g., polystyrene, acrylonitrile-butadiene-styrene (ABS), and polypropylene). Nowadays, the plant may process 90 kg/h of plastics with a low formation of char and gas and an interesting amount of liquid useful as a source of chemicals or fuel because it has an LHV of 35–43 kJ/kg. The Microwave-Assisted Pyrolysis (MAP) is an industrial novelty in plastic recycling, and it looks very promising for a much more modern and innovative plastic waste recovery system.",book:{id:"11145",title:"Recent Microwave Technologies",coverURL:"https://cdn.intechopen.com/books/images_new/11145.jpg"},signatures:"Marco Frediani, Piero Frediani, Gianni Innocenti, Irene Mellone, Roberto Simoni and Gianpaolo Oteri"},{id:"82420",title:"Applications of Microwaves in Medicine and Biology",slug:"applications-of-microwaves-in-medicine-and-biology",totalDownloads:0,totalDimensionsCites:0,doi:"10.5772/intechopen.105492",abstract:"This chapter deals with the description of recent research activities oriented on the perspective of microwave technologies in medicine and biology. It brings new ideas about the possibilities of using microwaves in thermotherapy—above all toward hyperthermia in cancer treatment. Development of new types of hyperthermia applicators (based, e.g., on technologies such as metamaterials, evanescent modes in waveguides, and other types of transmission structures) will be discussed here. Furthermore, we would like to underline in this chapter perspectives of microwaves in medical diagnostics. It is possible to expect that, e.g., microwave differential tomography, UWB radar, and microwave radiometers (all three can be used both for medical diagnostic and for noninvasive temperature measurement) will soon play an important role in it. Finally, experimental equipment necessary for research on the biological effects of EM fields is presented.",book:{id:"11145",title:"Recent Microwave Technologies",coverURL:"https://cdn.intechopen.com/books/images_new/11145.jpg"},signatures:"David Vrba, Jan Vrba, Ondrej Fiser, Jesus Cumana, Milan Babak and Jan Vrba Senior"},{id:"82158",title:"A Usability Analysis of the DAO Concept Based on the Case Study of a Blockchain Game",slug:"a-usability-analysis-of-the-dao-concept-based-on-the-case-study-of-a-blockchain-game",totalDownloads:3,totalDimensionsCites:0,doi:"10.5772/intechopen.105347",abstract:"Games based on a blockchain exist in all variants and facets, mostly as single- or multiplayer games. This chapter deals with the implementation of a multiplayer strategy game (Connect Four), using blockchain technology for decentralized data storage and the entire game logic. The focus is on the use of the decentralized autonomous organization (DAO) principle, for coordination and voting within the teams. The chosen game just stands as an example; other games or gamification approaches in which users can take decisions collaboratively can be used here. A web application was implemented acting as a central interface between players and the blockchain. Hence, it was possible for players to compete against each other in teams and to collectively decide the next move by participating in a roundly voting. With the help of a standardized questionnaire, answered by each player after each match, possible impacts of the voting mechanism on the usability were determined.",book:{id:"11552",title:"Gamification - Analysis, Design and Development",coverURL:"https://cdn.intechopen.com/books/images_new/11552.jpg"},signatures:"Lars Karbach, Mortiz Korte, Nils Orbat, Daniel Muschiol and March Jansen"},{id:"82415",title:"Power Consumption in CMOS Circuits",slug:"power-consumption-in-cmos-circuits",totalDownloads:2,totalDimensionsCites:0,doi:"10.5772/intechopen.105717",abstract:"In this chapter, we explain the two types of power consumption found in a complementary metal-oxide-semiconductor (CMOS) circuit. In general, a CMOS circuit tends to dissipate power at all times—be it active or inactive. The power consumed by the circuit when it is performing computational tasks is known as dynamic power. On the contrary, the power lost due to current leakage during which the circuit is dormant is referred to as static power. By carefully and properly designing the circuit, current leakage can be suppressed to its minimum. Hence, dynamic power consumption is usually significantly higher than its static counterpart. Some of the techniques that could be adopted to save dynamic power consumption include reducing the supply voltage, clock frequency, clock power, and dynamic effective capacitance. By probing into the activity factors of the design modules, the techniques can be applied to those with high power consumption.",book:{id:"11948",title:"Field-Effect Transistor",coverURL:"https://cdn.intechopen.com/books/images_new/11948.jpg"},signatures:"Len Luet Ng, Kim Ho Yeap, Magdalene Wan Ching Goh and Veerendra Dakulagi"}],onlineFirstChaptersTotal:797},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:8,limit:8,total:0},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:89,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:104,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:31,numberOfPublishedChapters:315,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:11,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:141,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:129,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:113,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:105,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:2,numberOfUpcomingTopics:1,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:5,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:14,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}},{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. 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\r\n\tEducation and Human Development is an interdisciplinary research area that aims to shed light on topics related to both learning and development. This Series is intended for researchers, practitioners, and students who are interested in understanding more about these fields and their applications.
",coverUrl:"https://cdn.intechopen.com/series/covers/23.jpg",latestPublicationDate:"June 25th, 2022",hasOnlineFirst:!0,numberOfPublishedBooks:0,editor:{id:"280770",title:"Dr.",name:"Katherine K.M.",middleName:null,surname:"Stavropoulos",slug:"katherine-k.m.-stavropoulos",fullName:"Katherine K.M. Stavropoulos",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRdFuQAK/Profile_Picture_2022-05-24T09:03:48.jpg",biography:"Katherine Stavropoulos received her BA in Psychology from Trinity College, in Connecticut, USA. Dr. Stavropoulos received her Ph.D. in Experimental Psychology from the University of California, San Diego. She completed her postdoctoral work at the Yale Child Study Center with Dr. James McPartland. Dr. Stavropoulos’ doctoral dissertation explored neural correlates of reward anticipation to social versus nonsocial stimuli in children with and without autism spectrum disorders (ASD). She has been a faculty member at the University of California, Riverside in the School of Education since 2016. Her research focuses on translational studies to explore the reward system in ASD, as well as how anxiety contributes to social challenges in ASD. She also investigates how behavioral interventions affect neural activity, behavior, and school performance in children with ASD. She is also involved in the diagnosis of children with ASD and is a licensed clinical psychologist in California. She is the Assistant Director of the SEARCH Center at UCR and is a Faculty member in the Graduate Program in Neuroscience.",institutionString:null,institution:{name:"University of California, Riverside",institutionURL:null,country:{name:"United States of America"}}},editorTwo:null,editorThree:null},subseries:{paginationCount:2,paginationItems:[{id:"89",title:"Education",coverUrl:"https://cdn.intechopen.com/series_topics/covers/89.jpg",isOpenForSubmission:!1,editor:{id:"260066",title:"Associate Prof.",name:"Michail",middleName:null,surname:"Kalogiannakis",slug:"michail-kalogiannakis",fullName:"Michail Kalogiannakis",profilePictureURL:"https://mts.intechopen.com/storage/users/260066/images/system/260066.jpg",biography:"Michail Kalogiannakis is an Associate Professor of the Department of Preschool Education, University of Crete, and an Associate Tutor at School of Humanities at the Hellenic Open University. He graduated from the Physics Department of the University of Crete and continued his post-graduate studies at the University Paris 7-Denis Diderot (D.E.A. in Didactic of Physics), University Paris 5-René Descartes-Sorbonne (D.E.A. in Science Education) and received his Ph.D. degree at the University Paris 5-René Descartes-Sorbonne (PhD in Science Education). His research interests include science education in early childhood, science teaching and learning, e-learning, the use of ICT in science education, games simulations, and mobile learning. He has published over 120 articles in international conferences and journals and has served on the program committees of numerous international conferences.",institutionString:"University of Crete",institution:{name:"University of Crete",institutionURL:null,country:{name:"Greece"}}},editorTwo:{id:"422488",title:"Dr.",name:"Maria",middleName:null,surname:"Ampartzaki",slug:"maria-ampartzaki",fullName:"Maria Ampartzaki",profilePictureURL:"https://mts.intechopen.com/storage/users/422488/images/system/422488.jpg",biography:"Dr Maria Ampartzaki is an Assistant Professor in Early Childhood Education in the Department of Preschool Education at the University of Crete. Her research interests include ICT in education, science education in the early years, inquiry-based and art-based learning, teachers’ professional development, action research, and the Pedagogy of Multiliteracies, among others. 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