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1. Introduction
Climate change alters environmental conditions and therefore has direct and biophysical effects on agricultural production. The biophysical and direct effects of climate change induce alterations on the prices and production of agriculture. Such changes are reflected on the economic system as farmers and other market participants make adjustments autonomously. They are both compelled to modify their crop combinations, use of supplies, level of production, and food demand, consumption and trade. Climate change causes a changes in rainfall regimes which have direct effects on crop yields as well as indirect effects through changes in the availability of water irrigation [1].
Sugarcane (Saccharum officinarum L.) is a monocotyledonous perennial plant belonging to the gramineous family Saccharum officinarum L. [2]. Sugarcane is a commercial crop in tropical and subtropical regions. According to FAOSTAT [3], sugarcane is cultivated in 26.1 million hectares producing 1.83 trillion canes. Sugarcane is a multi-purpose industrial cash crop and the main source of raw material for sugar production. It is responsible for almost 70% of world-produced centrifugal sugar [4]. Some mitigation and adaptation strategies for climate change in sugarcane production are the use of biotechnological techniques such as transcriptome, genetic transformation and in vitro micropropagation. In this chapter, we will talk about water stress in sugarcane caused by the climatic change and the biotechnological alternatives such as transcriptome, genetic transformation and micropropagation which are currently being carried out in our laboratory to counteract this problem.
2. Climate change, water stress and its effect in sugarcane
A large scale of plant production grown under different agricultural production systems is lost under the effects of abiotic stresses, which may result in a 70% reduction of the potential yields of crop plants [5]. During growth and developmental periods, crops suffer seasonal floods and droughts, extreme temperatures or salinity all year round. Globally, about 22% of global agricultural land is saline, and the increased damage caused by drought has been reported to limit plant growth and development followed by a loss of productivity, especially in crop species [6, 7]. Thus, drought stresses are one of the most serious kind of abiotic stresses that implies a threat on crop productivity worldwide.
Sugarcane, an important source of sugar and ethanol, is a relatively high water-demanding crop and its growth is highly sensitive to water deficit [8]. It is estimated that sugarcane produces 8–12 ton cane per ML of water irrigation [9], and water deficit can lead to productivity losses of up to 60% [10–13]. For this reason, production areas are concentrated in regions with favorable rain regime to sugarcane growth and development [14], while in other areas crop production requires supplemental or full irrigation [15].
According to various studies, water stress triggers many physiological, biochemical, and molecular responses that influence various cellular processes in plants and this impacts on its productivity [16, 17].
Severe water stress such as drought affects the entire plant. Morphological and physiological responses in sugarcane plants vary according to its genotype, duration (rapid or gradual) and intensity (severe or mild) of stress and also the type of affected tissue [18–21]. Water stress also affects both cane and sugar yield substantially. The most common water stress responses in sugarcane are leaf rolling, stomatal closure, inhibition of stalk and leaf growth, leaf senescence and reduced leaf area [12, 22]. Moreover, under water stress, both cell division and cell elongation are interrupted [23] and stem and leaf elongation are the most severely affected growth processes [24, 25]. Root development is also influenced by water deficit [19, 26] but its overall biomass is relatively less than the above-ground biomass. Sugarcane is a tropical crop with C4 photosynthetic metabolism. A moderate water stress causes a stomatal limitation, which triggers a decrease in stomatal conductance (gs), transpiration rate (E), internal CO2 concentration (Ci), and photosynthetic rate [26–30]. Under water stress, a decline in photosynthetic rate is mainly caused by a decrease in phosphoenolpyruvate carboxylase (PEPcase) and ribulose-1,5-biphosphate carboxylase (Rubisco) activity [26, 27, 31]. It is worth to note that photosynthesis rate is also impacted by sugar accumulation in leaves [32]. Under non-stressed condition low leaf sugar content is conducive to photosynthesis, while high sugar content moderates carbon fixation [33]. Interestingly, increased levels of some sugars, such as trehalose, can help plants to cope with water deficit, reducing the damage on cell membrane [34]. The capacity to accumulate trehalose was demonstrated in sugarcane roots under drought conditions. Sales et al. [35] reported an increase in starch hydrolysis, leading to higher levels of soluble sugars that helped sustain carbon supply even in a reduced CO2 fixation condition, facilitating growth recovery after stress.
3. Sugarcane and biotechnology
Sugarcane crop productivity has progressively increased to remarkable levels worldwide in the last century [36]. This increase in productivity has been ascribed to the development and widespread use of improved cultivars with increased resistance to diseases and pests, better management of water, nutrients and other resources, and the availability of relatively cheap chemical fertilizers and pesticides. Sustaining this pace of improvement in crop productivity by innovative and intensive agriculture, whilst ensuring minimal environmental impact, will be one of the major challenges to maintain a profitable sugar industry in the future.
Biotechnology offers excellent opportunities for sugarcane crop improvement. Commercial sugarcane, mainly the interspecific hybrids of S. officinarum and S. spontaneum [37], would greatly benefit from biotechnological improvements due to its complex polyploid-aneuploid genome, narrow genetic base, poor fertility, susceptibility to various diseases and pests, and the long duration (12–15 years) required to breed elite cultivars. More importantly, there is an ongoing need to provide durable disease and pest resistance commercial clones in combination with superior agronomic performance. This led to considerable research in different areas of biotechnology pertinent to sugarcane breeding and disease control. Despite the availability of molecular tools and strategies and advancements in our understanding of stress responses, engineering crops for drought tolerance remains a major challenge. This is not only due to the complexity of the plant responses to water deficit but also due to the difficulty of identifying and exploiting of large effect genes and alleles and the associated selection traits for developing drought tolerant varieties suitable for commercial crop production conditions [38].
4. Micropropagation an alternative to develop plants tolerant to water stress “hyperhydricity”
Various micropropagation systems such as liquid cultures and automation have proven the potential to resolve manual handling of in vitro cultures at various stages and decrease production cost. However, hyperhydricity is a major problem during in vitro culture of many crops in liquid culture systems. Hyperhydricity (also known as “vitrification”) is a physiological disorder occurring in plant material of tissue culture, which causes a reduction of propagation and death of tissues when transferred to ex vitro conditions [39–41]. The environment inside culture vessels normally used for plant micropropagation is characterized by high humidity, limited gaseous exchange between the internal atmosphere of the culture vessel and its surrounding environment, and the accumulation of ethylene; conditions that may induce physiological disorders [42]. The development of hyperhydric deformities represents a disadvantage for plant micropropagation and a barrier for the exploitation of bioreactor technologies to scale-up its production [41]. The concept of stress in relation to hyperhydricity is not completely established. Therefore, it remains difficult to assume when hyperhydric tissues are stressed. Previous studies argued that abnormal morphology observed in hyperhydricity could be attributed to changes occurring at cellular level due to the modifications of membrane composition or DNA content [42]. However, Rojas-Martínez and coworkers [41] considered this disorder as the result of the stressful conditions brought out by waterlogging of the apoplast. This causes hypoxia and thereby leads to severe oxidative stress. They concluded that hyperhydric features like vitreous appearance and wrinkled leaves are secondary events resulting from waterlogging of the apoplast.
The temporary immersion system (TIS) consists on the use of bioreactors with automated devices that control features such as gas exchange, liquid medium culture and lighting, required for the growth, development and survival of plants. TIS mainly consist of three phases: multiplication, elongation and rooting phase. Plantlets propagated in TIS have better performance than those propagated by conventional methods of micropropagation. TIS provides a rapid and efficient plant propagation system for many agricultural and forestry species, it utilizes liquid media avoiding intensive manual handling [43].
With the objective of evaluating the stress caused by hyperhydricity in the in vitro culture of sugarcane var. MEX69290, three types of culture were analyzed: Semisolid (Magenta) was used as control; Continuous immersion (250 ml Flask); and Temporary Immersion (BioMINT II Bioreactor). Multiplication, maturation, and ex vitro adaptation phases of sugarcane under these three types of culture were evaluated.
The obtained results in the adaptation of in vitro plants of S. officinarum at three different types of culture in the multiplication phase were surprising, as it is observed in Figure 1, where a notorious formation of shoots occurs in continuous immersion medium. Plants of var. MEX69290 obtained a much higher average shoot formation at the temporary immersion bioreactors than those observed in semi-solid medium. It was observed that invariable of the inoculum density applied (5, 10, 15 plants per bottle) was higher in continuous immersion. Similarly, growth index factor was higher in this culture system than that obtained in semi-solid medium or temporary immersion bioreactors (Figure 1). We can observe comparing our results with other works that the treatment response depends on the type of explant and variety of sugarcane. Several studies have reported that the rate of shoot formation is higher in temporary immersion bioreactors than in semi-solid cultures. It is important to mention that none of the previous works reported any problem with the hyperhydricity in the obtained in vitro plants. Only, Snyman [44] reports this condition on the induction and germination of somatic sugarcane embryos. Tesfa and coworkers [45], didn’t report problems of hyperhydricity or a decrease in field survival rate out of in vitro plants after using a liquid culture medium with agitation (80 rpm) in which they obtained an average shoot emission of 6.95 and 6.30 in the two cultivars used. The shoot emissions and growing index of the sugarcane variety MEX69290 was not affected when cultivated in a stationary liquid medium for 28 days (Figure 1).
Figure 1.
Average of shoots at different densities of inoculum. T5-T20: 5 inoculum plants were used in semi-solid and continuous immersion medium, 20 plants in temporary immersion; T10-T40: 10 inoculum plants were used in semi-solid and continuous immersion medium, and 40 plants in temporary immersion; T15-T60: 15 inoculum plants were used in semi-solid and continuous immersion medium, and 60 plants in temporary immersion; semisolid (red rectangle), continue immersion (orange rectangle) and temporary immersion (green rectangle). At the bottom of the figure, the calculated growth index factor is reported using the obtained fresh weight under the same inoculum density conditions; T5-T20 (red rectangle), T10-T40 (orange rectangle) and T15-T60 (green rectangle). Five replicates were carried out for each treatment.
The variety MEX69290 clones’ response at the maturation phase showed the same behavior as that observed at the multiplication phase, with the average shoot emission and the growth index being higher in the liquid culture than the one obtained in half semi-solid or in the temporary immersion bioreactor culture (Figure 2).
Figure 2.
Mean of shoots using 10 in vitro plants in semi-solid and continuous immersion cultures and 60 plants in temporary immersion bioreactors. At the bottom of the figure the calculated growth index factor is reported using the obtained fresh weight under the same inoculum density conditions. Five replicates were carried out for each treatment. Semisolid (red rectangle), continue immersion (orange rectangle) and temporary immersion (green rectangle).
After 28 days in maturation phase, 120 plants from semi-solid culture, 120 plants under continuous immersion, and 75 from BIOMINT were adapted. In Figure 3, we can observe the quality of the plants from the same clone at the three different cultivation systems.
Figure 3.
Phase adaptation of in vitro plants of S. officinarum var. MEX69290, seeded in a germination mixture BM2, previously autoclaved. 15 plants per container were adapted in growth culture room at 25°C with 16/8 hours photoperiod light/dark. (A) and (B) day zero and twenty-eight, of plants coming from semi-solid culture; (C) and (D) day zero and twenty-eight, of plants coming from liquid culture; (E) and (F) day zero and twenty-eight, of plants coming from temporary immersion system (BioMINT).
Plants underwent a 28 days preadaptation period, and afterward were planted and placed in greenhouse conditions. Once plants where transferred into the greenhouse, their survival rate was evaluated, being 100% in all cases (Figure 4). Plants from the temporary immersion bioreactors were taller and with longer leaves, but those from semi-solid medium and continuous immersion continued to emit shoots during the following 4 months evaluation at the greenhouse. The results obtained in this phase are very similar to those reported by Arencibia et al. [46], Bernal et al. [47], and Silva et al. [48], who reported survival rates higher than 96% in the different cultivars using a temporary immersion bioreactor, and our result is much higher than the studies reported by Snyman et al. [44], with only 34% of survival rate from sugarcane grown in the RITA system.
Figure 4.
Greenhouse adaptation of in vitro plants of S. officinarum var. MEX69290, from culture: (A) semi-solid; (B) continuous immersion; (C) temporary immersion. Substrate consisted on a 3: 1 mixture of sunshine: soil. All plantlets survived 100% after 30 days in the greenhouse.
The best results out of the measured parameters were obtained from the continuous immersion propagation system. It was concluded the reason for this may reside in the elimination of gelling agent, which additionally lowers production costs in the process of delivering this sugarcane’s variety to the field. Plants obtained under this system achieved normal development, they developed shoots and roots cyclically and no vitrification was detected in any of the evaluated micropropagation phases. This suggests that the clone obtained from the MEX69290 variety is tolerant to liquid culture conditions. Apparently this system does not generate an abiotic stress, stationing it as a prospective medium to perform genetic transformation processes and to study its gene expression pattern that could further make enhanced tolerant clones.
5. Transcriptomic analysis of an elite Mexican sugarcane cultivar (‘Mex 69-290’) in response to osmotic stress. Identification of genes with biotechnological potential
Modern sugarcane cultivars have been obtained by inter-specific hybridizations between the high-sucrose-yielding of S. officinarum (2n = 8x = 80) and the stress-tolerant S. spontaneum (2n = 40–128). As a consequence, sugarcane cultivars present large (10 Gb) and poly-aneuploid genomes with numerous gene alleles and repetitive sequences. Such genome complexity has made it difficult to obtain a complete sequenced reference genome that could aid in the identification of novel genes with biotechnological potential for the improvement of this important C4 crop. Alternatively, de novo transcriptome assembly of reads produced by high-throughput sequencing technologies (also referred to as Next Generation Sequencing (NGS)) offers a mean to unravel global gene expression changes in response to various conditions in sugarcane. For example, some recent works have employed High-throughput sequencing to identify sugarcane genes involved in leaf abscission [49], biomass content and composition [50], and abiotic stress [51]. Li and cols. [49] performed a transcriptome analysis to identify genes associated with leaf abscission in sugarcane. They employed the Illumina HiSeq 2000 platform (2x90pb) to analyze six cDNA libraries from parents and their F1 offspring, which present different leaf abscission behaviors. After a total assembly, they found 275,018 transcripts corresponding to 164,803 genes. Then, to identify genes related to leaf abscission in sugarcane [49], analyzed a core set of 1, 202 transcripts which were up-regulated in leaf abscission sugarcane plants (LASP) in comparison to leaf packaging sugarcane plants (LPSP). They found that some of these genes were associated with plant-pathogen interaction, response to stress, and ABA-associated pathways. On the other hand [50], performed an extensive transcriptome analysis to identify genes associated with biomass content. They employed the Illumina HiSeq 4000 platform to analyze cDNA libraries from 20 internodal samples of 10 different sugarcane genotypes, which were divided in low and high fiber containing groups. They found 5601 and 4659 unique expressed transcripts in High and Low fiber containing genotypes; and 83,421 shared expressed transcripts between both groups. Furthermore, they found 555 differentially expressed transcripts between low and high fiber containing genotypes. Of these, 151 and 23 transcripts corresponded to sugar and fiber accumulation, respectively. Some of these genes were involved in Carbohydrate metabolism, Photosynthesis, Cell-wall metabolism and Lignin Pathway; DIR proteins were also represented [50].
Regarding abiotic stress, Belesini and cols. [51] analyzed the transcriptomic profile of the drought-tolerant ‘SP81-3250’ and the drought-sensitive ‘RB855453’ sugarcane cultivars under drought stress conditions for 30, 60, and 90 days. They analyzed a total of 54 cDNA libraries by Illumina HiScanSQ System and HiSeq 2500 platforms. Among the genes that were induced in the drought-tolerant cultivar, they found an ascorbate peroxidase, a MYB TF, an E3 SUMO-protein ligase SIZ2, a coenzyme A ligase (a key enzyme for the biosynthesis of flavonoids), and an aquaporin, among others. These types of genes are well known to play a role in abiotic stress tolerance. In the drought-sensitive cultivar they found several kinases that were induced upon stress like Receptor like protein kinases (RLK), which might play a role in stress stimulus perception; bHLH transcription factors; ACC oxidase from the ethylene biosynthetic pathway; and many undescribed genes. More recently (2017), in our laboratory Pereira-Santana and cols. [52] analyzed the transcriptomic profile of the 2nd most important sugarcane cultivar in Mexico, ‘Mex 69-290’, in response to osmotic stress. In such study, authors employed the High-throughput sequencing system HiSeq-Illumina (2x100bp) to analyze 16 cDNA libraries representing leaves and roots of in vitro-grown plantlets exposed to PEG-8000 during 0, 24, 48, and 72 hours. After assembly of a total of 140, 339 unigenes, Pereira-Santana and cols. Found core sets of 536 and 750 up-regulated genes in response to osmotic stress in roots and leaves, respectively; and core sets of 1093 and 531 down-regulated genes in roots and leaves, respectively. After gene annotation, the authors found that sugarcane ‘MEX69290’ responds to osmotic stress by increasing the expression of genes involved in transcription regulation, oxide-reduction, carbohydrate catabolism, and flavonoid and other secondary metabolites biosynthesis. Genes responsive to ABA, water deprivation, and heat stress were also up-regulated. On the other hand, this sugarcane cultivar responds to osmotic stress by decreasing the expression of genes involved in sucrose and starch metabolic processes, cell wall biogenesis, cellulose biosynthesis, anion transport, and light response. A handful of the genes found by Pereria-Santana and cols. Are presented along with their expression profiles in the heat map of Figure 5A. Because of the well-defined expression pattern of some of these genes, they could prove to be useful as expression markers in the response of ‘MEX69290’ to osmotic stress. For example, ABA 8-hydroxylase 3, Isoflavone 2-hydroxylase, LEA 14A, and NAC TF 25 showed clear patterns of up-regulation. In fact, in our laboratory further expression and functional analyses are currently being carried out regarding this NAC TF25 gene. Conversely, Bidirectional sugar transporter SWEET11, Cellulose synthase E6, and Sugar transporter ERD6 16 showed clear patterns of down-regulation. These down-regulated genes are also interesting, not just because of their responsiveness to osmotic stress but also due to their involvement in sucrose metabolism. The engineering of these genes might increase biomass production in sugarcane and tolerance to osmotic stress simultaneously. Furthermore, many TFs known to play important roles in the stress responses of plants, i.e. HSF, ZN, bZIP, WRKY, NAC, and MYB, were found in abundance in the total assembly of the ‘MEX69290’ transcriptome (Figure 5B). Even when some of these TF families seemed underrepresented (like NAC and MYC), they still provide a useful benchmark to conduct phylogenetic, expression, and functional analysis.
Figure 5.
Selected DEGs in response to osmotic stress and abundance of major TF families and Dirigent protein family in sugarcane ‘MEX69290’ transcriptome. (A) Expression profile of 20 selected DEGs in leaves and roots of sugarcane ‘MEX69290’ plantlets submitted to PEG-8000 treatment during 0, 24, 48, and 72 hours. Data was obtained from the work of Pereira-Santana and cols [52]. The heat map was generated with the ComplexHeatmappackage v1.14.0 [52] in R v3.4.1 [53]. (B) Abundance of major stress-related TF families and Dirigent protein family in arabidopsis, rice, sorghum, and sugarcane. The results were obtained by means of HMM searches using the profiles of the HSF (PF00447), ZF (PF00096), bZIP (PF00170), WRKY (PF03106), NAC (PF02365), MYB (PF00249), and Dirigent (PF03018) proteins obtained from the Pfam database (http://pfam.xfam.org) [54]. For this analysis the complete predicted proteomes (primary transcripts only) of arabidopsis, rice, and sorghum were obtained from Phytozome v. 12 [55]. Sugarcane predicted protein dataset was obtained from the transcriptome assembly of Pereira-Santana and cols [52] HMM searches were performed using HMMER3 v3.1b2 (http://hmmer.org/) and set to a cut-off e-value of 1e-05 and a score above the inclusion threshold of each HMM profile.
In addition to the insights about the global gene expression dynamics of ‘Mex 69-290’ in response to osmotic stress and the identification of novel TFs, the work of Pereira-Santana and cols. Provides a useful benchmark for the study of other specific gene families of biotechnological significance for sugarcane engineering, for example the DIR protein family. Plant DIR proteins are believed to be involved in lignin biosynthesis, defense [56, 57], and abiotic stress responses such as dehydration [58], and salinity and oxidative stress [59]. In a recent study, 5 available sequence databases for sugarcane were surveyed, a total of 120 DIR proteins were identified [60]. Phylogenetic analysis showed that these DIR proteins are divided in 64 groups and 7 major clades: Dir-a, Dir-b/d, Dir-c, Dir-e, Dir-g, Dir-h, and Dir-i [60]. In the sugarcane transcriptome assembly of ‘sugarcane Mex 69-290’ performed in our laboratory by Pereira-Santana and cols, a total of 48 predicted proteins with DIR-like domains were identified. These DIR proteins were clustered in 7 groups according to their expression patterns (Figure 6). DIR42 protein from cluster 1 was significantly up-regulated in all time points of osmotic stress in root tissues. Conversely, DIR40 protein from cluster 7 was significantly down-regulated in all time points of osmotic stress in leaf tissues. In general, DIR genes from cluster 4 seem to possess a relative high expression in roots under control conditions, and those from cluster 7 seem to possess a relative high expression in leaves under control conditions. DIR genes from both clusters are down-regulated in response to osmotic stress. On the other hand, we also recovered a homolog of the ScDir gene (GenBank: JQ622282.1) from the sugarcane variety FN39 (DIR38 in cluster 5). The expression of ScDir from FN39 has been reported to be up-regulated in response to H2O2, NaCl, and PEG treatment [59]. Furthermore, its heterologous expression in Escherichia coli increases the bacterial host’s tolerance to NaCl and PEG [59]. The homolog of this gene in ‘Mex 69-290’ was slightly up-regulated in leaves, but down-regulated in roots (Figure 6, cluster 5). All of these mentioned DIR genes from sugarcane ‘MEX69290’ are interesting because they show differential expression patterns in leaves and roots in response to osmotic stress. However, their functional roles in osmotic stress tolerance and biomass accumulation still need to be experimentally analyzed. In summary, in the absence of a complete sequenced genome for sugarcane, high-throughput sequencing technologies applied to the elucidation of elite cultivars’ transcriptome profile are one of the most valuable resources for the identification of genes involved in both stress tolerance and biomass accumulation, which are important agronomic traits to face global climate change.
Figure 6.
Differential expression in response to osmotic stress of 48 Dirigent proteins found in sugarcane ‘MEX69290’ transcriptome. The 48 Dirigent sequences from sugarcane were grouped according to their expression profiles in 7 clusters (1–7). Data was obtained from Pereira-Santana and cols [52]. Heat map and sequence clustering were generated with ComplexHeatmap v1.14.0 [53] in R v3.4.1 [54] using the “euclidean” distance method and “complete” clustering method.
6. Genetic transformation of cane, a very powerful biotechnological tool to generate tolerant plants to water stress
According to the International Service for the Acquisition of Agri-biotech Applications (ISAAA), the worldwide distribution of genetically modified crops involves a total of 26 developing countries and 7 industrialized countries, headed by USA, Brazil, Argentina, Canada, India, China and South Africa. There is a current approval on the use of two commercial varieties of genetically modified cane in Brazil and Indonesia. On the former, plants containing the Cry1Ab gene, which produces an insecticidal toxin capable of killing the Diatraea caterpillar, are being cultivated. In Indonesia plants transformed with the EcBetA gene are resistant to drought.
Scientific research in genetic transformation have focused on resistance to biotic and abiotic factors such as weed control, production of renewable primary products, energy crops and production of pharmaceutically active substances.
Some of the methods in genetic transformation of plants are by Agrobacterium or biolistic which are time consuming, laborious and have low transformation efficiency. Thus we have attempted different options to optimize genetic transformation in sugar cane. An option for efficient transformation is by using different types of vectors, for example Anderson & Birch [61] used Binary super vectors in addition of different types of promoters (constitutive and inducible). Niu et al. [62] is other case who used the SoCINI inducible promoters and the ScMybRI constitutive promoters respectively [62, 63].
On the other hand, different in vitro culture protocols have been tried for decades to optimize the efficiency (time and management of the explant) as well as the number of transgenic plants. Yogesh and collaborators transformed cane leaves by Biolistic [64], regenerating seedlings via direct (ED) and indirect (EI) embryogenesis [65]. Arencibia and Carmona [66] reported genetic transformation by Agrobacterium tumefaciens and via indirect morphogenesis resulting in regenerated seedlings. Manickavasagam et al. reported regenerated seedlings after A. tumefaciens transformation via axillary shoots [67]. These latter two protocols require a time lapse between 3 and 6 months to generate seedlings.
In contrast, a genetic transformation protocol using A. tumefaciens has been developed in our laboratory (in the process of obtaining patent) where in vitro basal micro-shoots of MEX69290 cultivars underwent the insertion of the CpRap2.4b gene from the AP2/ERF transcription factor family, and out of cDNA of papaya stressed at 40°C. This genetic transformation protocol requires only 20 minutes and has a contamination rate of 0%, as well as a 21-day seedling regeneration rate. Our results showed a 70% survival in the first subculture and 100% in the second subculture with Kanamycin; similar results were reported by Manickavasagam regenerating transgenic seedlings using micro axillary outbreaks out of field plants [67], with a very laborious genetic transformation system and with 50% survival in the first crop. In addition, this work would be the second in sugarcane to report a gene of the AP2/ERF family of transcription factors inserted in sugar cane, the other work is the one reported by Reis et al. where they over expressed AtDREB2A CA (constitutive activity) in sugar cane [68]. In the transformed sugarcane seedlings generated by the genetic transformation protocol that was developed in our laboratory, the presence of the GFP was observed at the fluorescent emission of 395–475 nm, which indicates that the seedlings are transformed (Figure 7).
Figure 7.
GFP fluorescence of different plant leaves of sugar cane var. MEX69290. (A) Segment of wild leaf in visible light. (B) Wild leaf segment with emission at 509 nm. (C, E and G) Transgenic plants 1, 2 and 3 in visible light. (D, F and H) Transgenic plants 1, 2 and 3 with emission at 509 nm.
It should be clarified that the functionality of the CpRap2.4b gene belonging to the (AP2/ERF) transcription factors family was tested in tobacco plants, which were segregated to obtain F2 plants and were then subjected to water stress (drought) conditions to evaluate their function.
7. Conclusions
Climate change affects farmers economically, causing drought floods, which affect the productivity of the plant. Biotechnology is an alternative to reduce the impact of climate change on plants. In recent years there has been a continuing need to provide commercial clones of resistance to pests and long-lasting diseases in combination with superior agronomic performance. This led to considerable research in different areas of biotechnology including: micropropagation, transcriptomics and genetic transformation.
These areas of biotechnology together are a key tool in the pursuit of genetically enhanced plants that resist climate change.
Abbreviations
BAC
Bacterial artificial chromosome
EST
Expressed sequence tag
NGS
Next generation sequencing
GO
Gene Ontology
ABA
Abscisic acid
LEA
Late embryogenesis abundant
NAC
NAM, ATAF, and CUC
MYB
Myeloblastosis
HSF
Heat shock factor
ZF
Zinc Finger
TF
Transcription factor
ORF
Open reading frame
aa
Amino acids
DEG
Differentially expressed genes
HMM
Hidden Markov Model
TMM
Trimmed mean of M values
DIR
Dirigent
nt
Nucleotide
H2O2
Hydrogen peroxide
NaCl
Sodium chloride
PEG
Polyethylene glycol
CA
Constitutive activity
GFP
Green fluorescent protein
\n',keywords:"biotechnology, micropropagation systems, transcriptome, genetic transformation, sugarcane, abiotic stress",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/58418.pdf",chapterXML:"https://mts.intechopen.com/source/xml/58418.xml",downloadPdfUrl:"/chapter/pdf-download/58418",previewPdfUrl:"/chapter/pdf-preview/58418",totalDownloads:1014,totalViews:209,totalCrossrefCites:0,totalDimensionsCites:1,totalAltmetricsMentions:0,impactScore:0,impactScorePercentile:8,impactScoreQuartile:1,hasAltmetrics:0,dateSubmitted:"June 20th 2017",dateReviewed:"November 14th 2017",datePrePublished:"December 20th 2017",datePublished:"May 23rd 2018",dateFinished:"December 24th 2017",readingETA:"0",abstract:"Global climate change caused by natural processes results in major environmental issues that affect the world. Climate variability results in changes that cause water stress in plants. Sugarcane is a tropical grass C4, perennial and a multi-purpose industrial cash crop which serves as the main source of raw material for the production of sugar and biofuel. Farmers face the challenge to provide biotech alternatives with potential benefits and minimize potential adverse impacts on sugarcane’s production. In order to find biotechnology strategies to diminish the impact of climate change, our laboratory teamworks with micropropagation, transcriptome and genetic transformation of sugarcane using the var. MEX69290. In the transcriptome of sugarcane, a total of 536 and 750 genes were differentially regulated under normal and water stress treatment respectively, of which key genes were selected to be inserted into sugarcane for tolerance to abiotic stress. Regarding results of micropropagation, it was concluded that the continuous immersion propagation system was the best culture strategy. This may be as result of the elimination of gelling agent, which additionally helps reduce production costs.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/58418",risUrl:"/chapter/ris/58418",book:{id:"6377",slug:"plant-abiotic-stress-and-responses-to-climate-change"},signatures:"Evelyn Arlette Carrillo Bermejo, Miguel Angel Herrera Alamillo,\nSamuel David Gamboa Tuz, Miguel Angel Keb Llanes, Enrique\nCastaño de la Serna, Luis Manuel Robert Díaz and Luis Carlos\nRodríguez Zapata",authors:[{id:"62818",title:"Dr.",name:"Enrique",middleName:null,surname:"Castaño",fullName:"Enrique Castaño",slug:"enrique-castaao",email:"enriquec@cicy.mx",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"154564",title:"Dr.",name:"Luis",middleName:null,surname:"Rodriguez-Zapata",fullName:"Luis Rodriguez-Zapata",slug:"luis-rodriguez-zapata",email:"lcrz@cicy.mx",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"221080",title:"MSc.",name:"Samuel",middleName:null,surname:"Gamboa-Tuz",fullName:"Samuel Gamboa-Tuz",slug:"samuel-gamboa-tuz",email:"samuel.gamboa@cicy.mx",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"221083",title:"BSc.",name:"Evelyn Arlette",middleName:null,surname:"Carrillo-Bermejo",fullName:"Evelyn Arlette Carrillo-Bermejo",slug:"evelyn-arlette-carrillo-bermejo",email:"evelyn.carrillo@cicy.mx",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"221087",title:"MSc.",name:"Miguel Ángel",middleName:null,surname:"Herrera-Alamillo",fullName:"Miguel Ángel Herrera-Alamillo",slug:"miguel-angel-herrera-alamillo",email:"mianheal@cicy.mx",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"221088",title:"Dr.",name:"Manuel L.",middleName:null,surname:"Robert",fullName:"Manuel L. Robert",slug:"manuel-l.-robert",email:"robert@cicy.mx",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"226794",title:"MSc.",name:"Miguel A.",middleName:null,surname:"Keb-Llanes",fullName:"Miguel A. Keb-Llanes",slug:"miguel-a.-keb-llanes",email:"miguelke343@gmail.com",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Climate change, water stress and its effect in sugarcane",level:"1"},{id:"sec_3",title:"3. Sugarcane and biotechnology",level:"1"},{id:"sec_4",title:"4. Micropropagation an alternative to develop plants tolerant to water stress “hyperhydricity”",level:"1"},{id:"sec_5",title:"5. Transcriptomic analysis of an elite Mexican sugarcane cultivar (‘Mex 69-290’) in response to osmotic stress. Identification of genes with biotechnological potential",level:"1"},{id:"sec_6",title:"6. Genetic transformation of cane, a very powerful biotechnological tool to generate tolerant plants to water stress",level:"1"},{id:"sec_7",title:"7. Conclusions",level:"1"},{id:"sec_10",title:"Abbreviations",level:"1"}],chapterReferences:[{id:"B1",body:'Nelsona G, Valinb H, Sandsc R, Havlíkb P, Ahammadd H, Derynge D, et al. Climate change effects on agriculture: Economic responses to biophysical shocks. PNAS. 2014;111:3274-3279. DOI: 10.1073/pnas.1222465110'},{id:"B2",body:'Jahangir GZ, Nasir IA. Various hormonal supplementations activate sugarcane regeneration in vitro. Journal of Agricultural Science. 2010;2:231-237. DOI: 10.5539/jas.v2n4p231'},{id:"B3",body:'FAOSTAT. Sugarcane Production. Rome, Italy: Food and Agriculture Organization; 2014 [Internet]. Available from: http://faostat.fao.org/site/567/DesktopDefault.asp [Accessed: 21-02-2017]'},{id:"B4",body:'Sengar K. Developing an efficient protocol through tissue culture techniques for sugarcane micropropagation. 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DOI: 10.1007/s11103-005-2226-y'},{id:"B58",body:'Zhu L, Zhang X, Tu L, Zeng F, Nie Y, Guo X. Isolation and characterization of two novel dirigent-like genes highly induced in cotton (Gossypium barbadense and G. hirsutum) after infection by Verticillium dahliae. Journal of Plant Pathology. 2007;89:41-45'},{id:"B59",body:'Wu R, Wang L, Wang Z, Shang H, Liu X, Zhu Y, Deng X. Cloning and expression analysis of a dirigent protein gene from the resurrection plant Boea hygrometrica. Progress in Natural Science. 2009;19(3):347-352. DOI: 10.1016/j.pnsc.2008.07.010'},{id:"B60",body:'Jin-long G, Li-ping X, Jing-ping F, Ya-chun S, Hua-ying F, You-xiong Q, Jing-sheng X. A novel dirigent protein gene with highly stem-specific expression from sugarcane, response to drought, salt and oxidative stresses. Plant Cell Reports. 2012;31(10):1801-1812. DOI: 10.1007/s00299-012-1293-1'},{id:"B61",body:'Nobile PM, Bottcher A, Mayer JLS, Brito MS, dos Anjos IA, de Andrade Landell MG, Mazzafera P. Identification, classification and transcriptional profiles of dirigent domain-containing proteins in sugarcane. Molecular Genetics and Genomics. 2017;292(6):1323-1340. DOI: 10.1007/s00438-017-1349-6'},{id:"B62",body:'Anderson DJ, Birch RG. Minimal handling and super-binary vectors facilitate efficient, Agrobacterium-mediated, transformation of sugarcane (Saccharum spp. hybrid). Tropical Plant Biology. 2012;5:183-192. DOI: 10.1007/s12042-012-9101-1'},{id:"B63",body:'Niu J-Q, Wang A-Q, Huang J-L, Yang L-T, Li Y-R. Isolation, characterization and promoter analysis of cell wall invertase gene SoCIN1 from sugarcane (Saccharum spp.). Sugar Tech. 2014;17(1):65-76. DOI: 10.1007/s12355-014-0348-8'},{id:"B64",body:'Yogesh T, Maria G, Fredy A. Comparison of direct and indirect embryogenesis protocols, biolistic gene transfer and selection parameters for efficient genetic transformation of sugarcane. Plant Cell, Tissue and Organ Culture (PCTOC). 2012;111:131-141. DOI: 10.1007/s11240-012-0177-y'},{id:"B65",body:'Zale J, Jung JH, Kim JY, Pathak B, Karan R, Liu H, Chen X, Wu H, Candreva J, Zhai Z, Shanklin J, Altpeter F. Metabolic engineering of sugarcane to accumulate energy-dense triacylglycerols in vegetative biomass. Plant Biotechnology Journal. 2016;14:661-669'},{id:"B66",body:'Arencibia AD, Carmona ER. Sugarcane (Saccharum spp.). In: Wang K, editor. Agrobacterium Protocols. 2nd ed. Totowa, NJ: Humana Press; 2006. pp. 227-235. DOI: 10.1385/1-59745-131-2:227'},{id:"B67",body:'Manickavasagam M, Ganapathi A, Anbazhagan VR, Sudhakar B, Selvaraj N, Vasudevan A, Kasthurirengan S. Agrobacterium-mediated genetic transformation and development of herbicide-resistant sugarcane (Saccharum species hybrids) using axillary buds. Plant Cell Reports. 2004;23:134-143. DOI: 10.1007/s00299-004-0794-y'},{id:"B68",body:'Reis RR, da Cunha BADB, Martins PK, Martins MTB, Alekcevetch JC, Chalfun-Júnior A, et al. 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The Portuguese Experience",slug:"participatory-plant-quality-breeding-an-ancient-art-revisited-by-knowledge-sharing-the-portuguese-ex",signatures:"Maria Carlota Vaz Patto, Pedro Manuel Mendes-Moreira, Mara Lisa\nAlves, Elsa Mecha, Carla Brites, Maria do Rosário Bronze and Silas\nPego",authors:[{id:"70071",title:"Prof.",name:"Maria",middleName:null,surname:"Bronze",fullName:"Maria Bronze",slug:"maria-bronze"},{id:"160017",title:"Dr.",name:"Maria Carlota",middleName:null,surname:"Vaz Patto",fullName:"Maria Carlota Vaz Patto",slug:"maria-carlota-vaz-patto"},{id:"160020",title:"MSc.",name:"Pedro",middleName:null,surname:"Mendes-Moreira",fullName:"Pedro Mendes-Moreira",slug:"pedro-mendes-moreira"},{id:"160021",title:"Dr.",name:"Carla",middleName:null,surname:"Brites",fullName:"Carla Brites",slug:"carla-brites"},{id:"160022",title:"Dr.",name:"Silas",middleName:null,surname:"Pego",fullName:"Silas Pego",slug:"silas-pego"},{id:"166078",title:"MSc.",name:"Mara Lisa",middleName:null,surname:"Alves",fullName:"Mara Lisa Alves",slug:"mara-lisa-alves"},{id:"166079",title:"MSc.",name:"Elsa",middleName:null,surname:"Mecha",fullName:"Elsa Mecha",slug:"elsa-mecha"}]}]}],publishedBooks:[{type:"book",id:"597",title:"Crop Production Technologies",subtitle:null,isOpenForSubmission:!1,hash:"7f87c31dfd7e38f3e10cf7ec02df2201",slug:"crop-production-technologies",bookSignature:"Peeyush Sharma and Vikas Abrol",coverURL:"https://cdn.intechopen.com/books/images_new/597.jpg",editedByType:"Edited by",editors:[{id:"73200",title:"Dr.",name:"Peeyush",surname:"Sharma",slug:"peeyush-sharma",fullName:"Peeyush Sharma"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1248",title:"Resource Management for Sustainable Agriculture",subtitle:null,isOpenForSubmission:!1,hash:"3445faea41382312cdb15ca628229a20",slug:"resource-management-for-sustainable-agriculture",bookSignature:"Vikas Abrol and Peeyush Sharma",coverURL:"https://cdn.intechopen.com/books/images_new/1248.jpg",editedByType:"Edited by",editors:[{id:"136230",title:"Dr.",name:"Vikas",surname:"Abrol",slug:"vikas-abrol",fullName:"Vikas Abrol"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1404",title:"Aflatoxins",subtitle:"Detection, Measurement and Control",isOpenForSubmission:!1,hash:"e3a2b9bd1c46dd47875d6a0f3d8b2a39",slug:"aflatoxins-detection-measurement-and-control",bookSignature:"Irineo Torres-Pacheco",coverURL:"https://cdn.intechopen.com/books/images_new/1404.jpg",editedByType:"Edited by",editors:[{id:"62984",title:"Dr.",name:"Irineo",surname:"Torres-Pacheco",slug:"irineo-torres-pacheco",fullName:"Irineo Torres-Pacheco"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"2030",title:"Research in Organic Farming",subtitle:null,isOpenForSubmission:!1,hash:"11136b8e73780b3196ddb1ef0ce1571e",slug:"research-in-organic-farming",bookSignature:"Raumjit Nokkoul",coverURL:"https://cdn.intechopen.com/books/images_new/2030.jpg",editedByType:"Edited by",editors:[{id:"87654",title:"Dr.",name:"Raumjit",surname:"Nokkoul",slug:"raumjit-nokkoul",fullName:"Raumjit Nokkoul"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"2079",title:"Problems, Perspectives and Challenges of Agricultural Water Management",subtitle:null,isOpenForSubmission:!1,hash:"183bb777195754e887da67131255661f",slug:"problems-perspectives-and-challenges-of-agricultural-water-management",bookSignature:"Manish Kumar",coverURL:"https://cdn.intechopen.com/books/images_new/2079.jpg",editedByType:"Edited by",editors:[{id:"102967",title:"Dr.",name:"Manish",surname:"Kumar",slug:"manish-kumar",fullName:"Manish Kumar"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}],publishedBooksByAuthor:[]},onlineFirst:{chapter:{type:"chapter",id:"70121",title:"An Electrochemical Sensor Based on Electroreduction of Graphene Oxide on a Glassy Carbon Electrode Using Multiple Pulse Amperometry for Simultaneous Determination of l-Dopa and Benserazide",doi:"10.5772/intechopen.89685",slug:"an-electrochemical-sensor-based-on-electroreduction-of-graphene-oxide-on-a-glassy-carbon-electrode-u",body:'\n
\n
1. Introduction
\n
Parkinson’s disease (PA) is an illness that affects about 1% of the world population, according to the World Health Organization (WHO), and this number tends to increase considerably, as demonstrated by recent studies conducted by the University of Rochester. Its estimated that the number of people affected by the PA in the 15 countries analyzed will more than double up to 2040 [1]. Thus, it becomes extremely important to develop research on this disease and the treatment employed. Mainly alternative routes for the quantification of compounds are present in the drug used in the treatment of BP, which provides an effective and lower cost treatment.
\n
Parkinson’s disease is a condition that attacks the central nervous system (CNS) and the brain, and this evil affects the amount of dopamine in the body. Dopamine (DA) is an existing neurotransmitter catecholamine in the CNS and in the mammalian brain. It has a vital role in maintaining the activities of the central nervous system, cardiovascular system, and hormonal system [2]. Patients suffering from BP have a considerable decay in the production OF DA in the brain. The medications for this disease have the design of elevating the dopamine index in the brain [3].
\n
However, it is not possible to inject dopamine directly into the patient, because the blood-brain barrier does not allow the arrival of this hormone in the encephalon. With this, l-Dopa (LD) is used, a medicine that can overcome such a barrier and is converted into the encephalon in DA [4, 5]. Despite this, when there is an irregularity in the levels OF DA, there are some side effects such as nausea, vomiting, cardiac arrhythmia, schizophrenia, and dyskinesia [2, 6].
\n
Unfortunately, Dopa decarboxylase (DDC) quickly converts LD into the bloodstream, so only a small percentage reaches the brain. By inhibiting the enzyme, higher amounts of l-Dopa administered can reach the brain [7]. In order not to be converted “early,” the drug hydrochloride is introduced in the benserazide (BZ), which acts as a DDC inhibitor [8].
\n
Electroanalytical methods emerged as an alternative line when compared to chromatographic methods that demand the use of expensive instruments, which are experimentally complex and require a lot of time to analyze with various pretreatments of the sample. These methods are based on the different electroanalytical and electrochemical techniques. These in turn have excellent sensitivity, selectivity, speed, reduced costs, and possibility of miniaturization of the system, making the electrochemical sensors a promising tool to supplement the existing techniques. Therefore, the development of new strategies with the aim of perfecting and improving the electrochemical techniques is promising [9, 10, 11].
\n
Solid electrodes of different materials are employed in the electroanalytical methods; one of the most widely used materials is those based on carbonaceous materials, such as carbon graphite, pyrolytic carbon, and glass carbon. In addition to serving as a base electrode, carbonaceous materials are also used as modifiers, in order to catalyze the redox process and/or increase selectivity. Some examples that can be found in the literature are the use of reduced graphene oxide (RGO), graphene, nanotubes (single or multiple wall), etc.
\n
For this work the glass carbon electrode (GCE) was chosen as the base electrode and the reduced graphene oxide (rGO) by electroreduction. The GO could be reduced also by thermal and chemical process; however the electrochemical reduction causes a homogeneity and stability of the surface of the work electrode; for this reason we opted for the use of electroreduction of GO.
\n
The process of reduction of GO by electrochemical route is described in the literature in two ways, one using conventional amperometry (Amp) and the other using cyclic voltammetry (CV). The methodology employing the Amp consists in the application of a single potential in a specific time range, already the methodology employing CV focused on the application of a potential window at different scanning speeds and number of cycles.
\n
In order to add value to this work, the use of multiple pulse amperometry (MPA) was tested, a technique derived from conventional amperometry, and there are still no reports in the literature of this technique used for the purpose of reducing GO. Since MPA consists in the application of different potentials in a certain time range, this process has as an advantage the application of activation potentials of the electrode surface and cleaning [12]. Therefore, these possibilities can generate better efficiency in the reduction process; the number of pulses of potential applicable considering the software GPES reaches 10 and has the possibility of acquiring the current as a function of time to each potential pulse.
\n
The glass carbon electrode has distinct properties of its allotropic forms, pyrolytic graphite, and diamond carbon, which are also used in electrochemistry. However, it presents the best synergism of physical properties, affinity with the modifier material, and its good affinity with the selected immobilization methodology, which is a prerequisite described in the literature [13].
\n
The material that will carry out the chemical modification of the base electrode will be the GO, which will be reduced electrochemically in order to make it a conductive material, thus decreasing the resistivity of the electrode facilitating the transfer of electrons. The GO consists of a graphene sheet, a carbon structure consisting of sp2 bonds, which has these connections transformed into sp3 bonds by the substitutes groups, which removes its conductive characteristic. The reduction process causes this material to lose some functional groups and return to conductive characteristics and maintain the interaction from the functional groups [14, 15, 16, 17].
\n
The proposal of this work is the development of an electroanalytical method for the electrochemical reduction of graphene oxide and the simultaneous determination of LD and BZ in an electrochemical cell of three electrodes.
\n
\n
\n
2. Construction of an electrochemical sensor based on electroreduction of graphene oxide on a GCE
\n
\n
2.1 Instrumentation
\n
Electrochemical measurements and the formation of the work electrode modifier film were performed in a multi-potentiostat/galvanostat model PGSTAT101 coupled to a microcomputer containing the new 1.11 software and a microcomputer containing the GPES 4.1 software. As a system for obtaining electrochemical measurements, an electrochemical cell containing three electrodes was used: a saturated Ag/AgCl electrode containing 3.0 mol L−1 of KCl as a reference electrode, a platinum wire as an auxiliary electrode, and the GCE/rGO as a working electrode.
\n
Sodium phosphate dibasic heptahydrate, monobasic sodium phosphate monohydrate obtained from the Synth®, phosphoric acid, and 3.4-dihydroxy-l-phenylalanine were used; benserazide hydrochloride is acquired from Sigma-Aldrich®. All the chemical reagents used were analytical grade, and the buffer solutions were prepared with purified water by Millipore Milli-Q system.
\n
The solution containing l-Dopa was prepared with a solution of phosphoric acid pH 2.00 and concentration 0.1 mol L−1, and the solution of benserazide was prepared in phosphate buffer pH 5.50 and concentration 0.1 mol L−1.
\n
\n
\n
2.2 Pretreatment of glassy carbon electrode
\n
The glassy carbon electrode was pretreated by mechanical polishing with Alumina, HNO3, and H2O and also by electrochemical activation performed in a 0.5 mol L−1 H2SO4 solution for 20 scans at scan rate of 100 MV s−1 and a potential range −0.2 to 1.6 V.
\n
\n
\n
2.3 Electroreduction of graphene oxide
\n
The graphene oxide (GO) was synthesized based on previous works published [14]. The dispersion of GO was prepared in ethanol and Nafion 5% v/v. The construction of the proposed sensor was carried out by adding 5 μL of the GO suspension in the GCE. Then the electrode was kept in the oven at 60°C for 10 minutes. The electrochemical reduction of the GO was performed at multiple pulses to obtain the rGO. The reduction was carried out using a three-electrode setup: reference electrode (Ag/AgCl), auxiliary electrode (platinum wire), and working electrode (GCE) at PBS 0.1 mol L−1 and pH 7.00. \nFigure 1\n summarizes the process of modifying of the GCE.
\n
Figure 1.
Process of modifying the surface of the glassy carbon electrode.
\n
\n
\n
2.4 Multiple pulse amperometry
\n
The electrochemical reduction of rGO by MPA is not described in the literature. However, based on the procedure via amperometry and by cyclic voltammetry [18, 19, 20, 21, 22], some conditions were adopted as a starting point. According to the literature review, [22] the application of a negative potential after the reduction process using cyclical voltammetry tends to improve the reduction and homogeneity of the film, with this in mind and in order to enjoy the best performance possible of the MPA technique was adopted 9 potentials for the process of reduction of GO, optimizing which would be these potentials, the time of application and the concentration of GO.
\n
Considering that mechanisms may change according to the conditions used in electroreduction, some experimental parameters will be evaluated in the MPA reduction process. Initially, four parameters were optimized—applied potential, pulse application time, GO concentration, and pH effect. The objective of the optimization of these parameters is to develop a sensor with higher sensitivity in the determination of LD and BZ, and the conditions and results obtained in the optimization process will be discussed later.
\n
\n
\n
\n
3. Simultaneous determination of l-Dopa and benserazide
\n
\n
3.1 Electrochemical behavior of l-Dopa and benserazide
\n
The electrochemical behavior of LD against different sensor configurations, bare GCE and GCE/rGO before being optimized and after optimization, is shown in \nFigure 2A\n. An associated oxidation peak around 0.45 V is observed from the oxidation of hydroxyls bound to the LD aromatic ring. This oxidation process involves two electrons, and it is a similarly reversible process [23, 24, 25]. The cyclic voltammogram obtained with GCE/rGO after optimizing exhibits a current variation of approximately 1300% over simple GCE, as shown in \nFigure 2B\n. The observed current gain for l-Dopa is also observed in benserazide, thus making rGO a viable material for GCE modification.
\n
Figure 2.
LD 0.1 μmol L−1 electrochemical behavior in the analytical response (A) GCE, GCE/rGO before optimization, and GCE/rGO after optimization (B).
\n
The increase in current observed in the oxidation process of LD and BZ is due to some characteristics of rGO. Reduced graphene oxide consists of a graphene sheet with the presence of some functional groups along its structure, which will make the interaction between the base electrode and the analyte easier. In addition, rGO is a conductive material thus facilitating the exchange of electrons between the analyte and the WE.
\n
It is noteworthy that the increase in observed current from the addition of rGO leads to an increase in the linear working range of the sensor and an increase in its sensitivity.
\n
\n
\n
3.2 Optimization of experimental parameters
\n
\n
3.2.1 Effect of applied potential range
\n
Initially we analyzed potential range to reduce the graphene oxide. These potentials were in the window of −0.50 to −1.40 V. \nFigure 3\n indicates the behavior of the analytes before the different potentials adopted, and the anodic current was adopted as the selection method of the best result and also took into account the separation of current peaks anodic of LD and BZ. This figure does not represent the process of reducing but the electrochemical behavior of analytes using the GCE/rGO after the reduction.
\n
Figure 3.
(A) Cyclic voltammograms obtained at different electrodes prepared via MPA with different potentials (PBS 0.1 mol L−1, pH 5.50) after addition of 1.0 × 10−5 mol (BZ) and 9.95 × 10−5 mol L−1 (LD). (B) Analytical response.
\n
Among the potentials adopted, (−0.50, −0.60, −0.70, −0.80, −0.90, −1.00, −1.10, − 1.20, −1.30), (−0.60, −0.70, −0.80, −0.90, −0.95, − 1.00, −1.10, − 1.20, −1.30), and (−0.70, −0.80, −0.90, −1.00, −1.05, −1.10, −1.20, −1.30, −1.40), the range of −0.60 to −1.30 V presented the best result; therefore this value was adopted as the optimum value, and the analysis process continued.
\n
\n
\n
3.2.2 Effect of time
\n
After determining the potentials to be applied in the electroreduction, the time of application of these potentials was evaluated. Three application times were analyzed: 400, 450, and 500. \nFigure 4\n represents the voltammograms obtained after the GO reduction process, and this analysis was performed in the presence and absence of analyte for electrodes prepared at different times of reduction.
\n
Figure 4.
(A) Cyclic voltammograms obtained at different electrodes prepared via MPA with different times (PBS 0.1 mol L−1, pH 5.50) after addition of 1.0 × 10−5 mol (BZ) and 9.95 × 10−5 mol L−1 (LD). (B) Analytical response.
\n
Through the current difference, it was determined which would be the best fear of the application of the pulses, having as optimum time 450 s.
\n
\n
\n
3.2.3 Effect of GO concentration
\n
With the potentials and time of application well-defined, the evaluation of the GO concentration added to the electrode surface was performed, from 0.5 to 3 mg ml−1. \nFigure 5\n shows the voltammograms for the different concentrations of the solutions added on the GCE, and the optimum condition was defined by means of the current difference. This condition was 2.00 mg mL−1.
\n
Figure 5.
(A) Cyclic voltammograms obtained at different electrodes prepared via MPA with different GO concentrations (PBS 0.1 mol L−1, pH 5.50) after addition of 1.0 × 10−5 mol (BZ) and 9.95 × 10−5 mol L−1 (LD). (B) Analytical response.
\n
\n
\n
3.2.4 Effect of pH
\n
Finally, the pH influence of the electrolytic solution in the reduction process was evaluated, since it can cause interferences in the process of electroreduction, such as damage to the modifier material. Concerning this study, the optimal measurements previously evaluated in \nFigure 6\n can be observed, and the influence of the pH value of the support solution—phosphate buffer at 0.10 mol L−1—in the formation of the rGO film. By means of the current difference, the best pH value was defined, which was 6.00.
\n
Figure 6.
(A) Cyclic voltammograms obtained at different electrodes prepared via MPA with different pH (PBS 0.1 mol L−1, pH 5.50) after addition of 1.0 × 10−5 mol (BZ) and 9.95 × 10−5 mol L−1 (LD). (B) Analytical response.
\n
It should be noted that when the dispersion concentration of the modified material was optimized due to the influence of the thickness of the modification and the number of functional groups, and the thicker and the smaller the number of groups, the lower the number of functional loads will be transported. The potential is applied to eliminate the excess of functional groups, transforming the GO into or returning to give conductive characteristics of this material. The reduction time in turn allows a better interaction of the film with the reduction process making its surface more homogeneous.
\n
Therefore, it is extremely important that there is an optimization of the parameters of the technique and the concentration of modifier material to obtain a more efficient sensor, that is, with good sensitivity and selectivity.
\n
\n
\n
\n
3.3 Optimization of experimental conditions
\n
After the optimization of the GO reduction process, it was evaluated which of these presented better conditions for the continuation of the analyses of LD and BZ. The choice of the best technique was based on the separation of the peaks of LD and BZ and the higher anodic current.
\n
\n
3.3.1 Influence of ionic force
\n
It is known that the ionic force is directly linked to the feasibility of the analysis once the concentration of the ions influences in the transport of loads, which can generate an increase in the signal of the analyte.
\n
The determination of the ionic strength of the solution was made by means of the difference of white current and the addition of the analytes obtained in a buffer system pH 5.50, and the electrolyte concentration support was varied from 0.025 to 0.200 mol L−1. \nFigure 7\n illustrates the voltammograms obtained for these different concentrations, and through them it was possible to define that the ionic force that best suited was 0.050 mol L−1. The higher concentration may be happening with the competition between the electrolyte ions support and the analytes and concentrations below these there are not enough ions to carry out the transport of loads.
\n
Figure 7.
(A) Cyclic voltammograms obtained at different electrodes prepared via MPA with different electrolyte concentration (PBS pH 5.50) after addition of 1.0 × 10−5 mol (BZ) and 9.95 × 10−5 mol L−1 (LD). (B) Analytical response.
\n
\n
\n
3.3.2 pH influence
\n
Subsequently, the pH effect was evaluated, which may influence the electrode stability and provide secondary reactions of the analytes, such as LD, which in basic ph suffers secondary reactions which degrade it. This parameter was also evaluated by analyzing anodic current, and the higher these differences are the signal obtained, \nFigure 8\n illustrates the voltammograms for the different pH values of the electrolyte support, the optimum value for analysis was determined being 5.50.
\n
Figure 8.
(A) Cyclic voltammograms obtained at different electrodes prepared via MPA with different pH of the electrolyte (PBS 0.050 mol L−1) after addition of 1.0 × 10−5 mol (BZ) and 9.95 × 10−5 mol L−1 (LD). (B) Analytical response.
\n
\n
\n
\n
3.4 Calibration curve
\n
First, we calculated the amount of analyte needed to prepare a solution containing LD and BZ in the same proportion found in Prolopa®, from 5.23 mol of LD to 1.00 mol of BZ, which would be analyzed. Through this solution, the voltammetry analysis was performed using proposed sensor (previously optimized) as shown in \nFigure 9\n. The analytical curves were constructed for LD (\nFigure 10\n) and BZ (\nFigure 11\n).
\n
Figure 9.
Voltammograms (PBS 0.05 mol L−1 pH 5.5) and calibration curve, using cyclic voltammetry, containing LD in the concentration range of 2.53 × 10−5 mol L−1 to 4.19 × 10−4 mol L−1 and BZ of 4.83 × 10−6 mol L−1 to 8.02 × 10−5 mol L−1.
\n
Figure 10.
Calibration curve of BZ in phosphate buffer, pH 5.50, and concentration 0.05 mol L−1.
\n
Figure 11.
Calibration curve of LD in phosphate buffer, pH 5.50, and concentration 0.05 mol L−1.
\n
Having all the data optimized, it was possible to construct the final analytical curves by means of successive additions of LD 5.08 mmol L−1 and BZ 0.97 mmol L−1 in order to obtain the sensibility, the detection limit (LOD), and the limit of quantification (LOQ) for each analyte.
\n
The sensibility for LD was 0.05148 μmol L−1 and 0.10303 μmol L−1 for BZ. For the analytical curve, to have an acceptable degree of reliability, the value of R must be close to 1. The values obtained for R are above 0.98, which indicates a considerable good degree of reliability.
\n
The detection limit (LOD) represents the smallest amount of the analyte present in a sample that can be detected by the method. From the parameters of the analytical curve, the LOD can be expressed by LOD = 3 × (S/B), where S is the standard deviation of the white and B is the slope of the analytical curve.
\n
The limit of quantification (LOQ) is the smallest amount of the analyte in a sample that can be determined with accuracy and accuracy acceptable under the established experimental conditions. The LOQ is established through the analysis of samples containing decreasing concentrations of the analyte up to the lowest determinable level and can be expressed by the equation: LOD = 10 × (S/B), where S and B have the same values found for the LOD, previously.
\n
The limit of detection and the limit of quantification of LD and BZ obtaining was, respectively, 5.14 μmol L−1 and 17.10 μmol L−1 for LD and 8.96 × 10−1 μmol L−1 and 2.99 μmol L−1 and BZ. However, the detection limit can be adopted as the first point of the analytical curve, thus becoming 4.83 μmol L−1 for BZ and 25.30 μmol L−1 for LD.
\n
After the calibration curve was constructed, reproducibility and repeatability tests were performed. Reproducibility was confirmed by inter-day testing, where analyses of the same sample were performed on different days, and by intra-day testing, where five electrodes were built and samples of the same concentration were analyzed, after these tests. A good reproducibility must have standard deviation of less than 5% which will occur.
\n
Repeatability can be ratified by inter- and intra-day analysis, and it was made by intra-day analysis, where an electrode was built to analyze five samples of the same concentration, which presented a standard deviation below 5%, which statistically proves the repeatability of the method.
\n
It is noteworthy that all analyses presented throughout the text were made in triplicate, where a relative error of less than 5% was observed, which statistically proves that the analyses presented are within an acceptable standard.
\n
\n
\n
3.5 Application in pharmaceutical formulation
\n
In order to evaluate the applicability of the proposed method, a solution of LD and BZ was prepared employing the sample of the drug (Prolopa®) at the concentration of 5.08 mmol L−1 of LD and 0.97 mmol L−1 of BZ, having the nominal value of each analyte present in the Prolopa® as a reference for calculating the indicated concentrations. This solution was analyzed by the proposed sensor.
\n
Then, the recovery tests were performed. The results were obtained in triplicate and are presented in \nTables 1\n and \n2\n.
\n
\n
\n
\n
\n
\n
\n\n
\n
Measure
\n
[LD] added
\n
[LD] recovered
\n
Recovery (%)
\n
ER (%)
\n
\n\n\n
\n
1
\n
2.53 × 10−5 mol L−1\n
\n
2.60 × 10−5 mol L−1\n
\n
102.77
\n
2.77
\n
\n
\n
2
\n
2.87 × 10−4 mol L−1\n
\n
2.78 × 10−4 mol L−1\n
\n
96.86
\n
−3.14
\n
\n
\n
3
\n
4.19 × 10−4 mol L−1\n
\n
3.99 × 10−4 mol L−1\n
\n
97.91
\n
−2.09
\n
\n\n
Table 1.
Results of the LD addition and recovery tests in a solution prepared with the drug.
\n
\n
\n
\n
\n
\n
\n\n
\n
Measure
\n
[BZ] added
\n
[BZ] recovered
\n
Recovery (%)
\n
ER (%)
\n
\n\n\n
\n
1
\n
4.83 × 10−5 mol L−1\n
\n
4.95 × 10−5 mol L−1\n
\n
102.48
\n
2.48
\n
\n
\n
2
\n
5.50 × 10−5 mol L−1\n
\n
5.25 × 10−5 mol L−1\n
\n
95.45
\n
−4.55
\n
\n
\n
3
\n
8.02 × 10−5 mol L−1\n
\n
8.41 × 10−5 mol L−1\n
\n
104.86
\n
4.86
\n
\n\n
Table 2.
Results of the addition and recovery tests of BZ in a solution prepared with the drug.
\n
Through the results presented in \nTables 1\n and \n2\n, it was possible to observe that the sensor proved to be promising in the simultaneous determination of the analytes with a maximum relative error of ±5%. And the results found were analyzed in the same system with successive additions of the drug solution.
\n
It is noteworthy that the methodology of modification of the surface of the GCE is rapid and that the data obtained when compared with those present in the literature [23, 26, 27, 28, 29, 30, 31, 32, 33] has performance as good as the sensors described; however, the preparation methodology of some is more laborious than the proposal in this work.
\n
\n
\n
\n
4. Conclusion
\n
The proposed methodology was successfully developed, since it proved to be effective in the simultaneous determination of LD and BZ. It is worth noting that the methodology employed uses a rapid analysis technique, when comparing the separation techniques (chromatography).
\n
In short, the work proved to be efficient and innovative when compared to the literature but can have the methodology of reduction by MPA improved. Thus, this study enable an improvement in the methodology of analysis and the description of this technique with the purpose of electroreduction and/or electropolymerize in the literature.
\n
\n
Acknowledgments
\n
We thank FAPEMIG, CAPES, CNPQ and Rede Mineira de Química for the continuous support of our research.
\n
\n',keywords:"graphene oxide, electroreduction, simultaneous determination, l-Dopa, benserazide",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/70121.pdf",chapterXML:"https://mts.intechopen.com/source/xml/70121.xml",downloadPdfUrl:"/chapter/pdf-download/70121",previewPdfUrl:"/chapter/pdf-preview/70121",totalDownloads:481,totalViews:0,totalCrossrefCites:0,dateSubmitted:"June 10th 2019",dateReviewed:"September 11th 2019",datePrePublished:"November 20th 2019",datePublished:"May 6th 2020",dateFinished:"November 20th 2019",readingETA:"0",abstract:"In this work, we described the development an electrochemical sensor based on electroreduction of graphene oxide (rGO) on a glassy carbon electrode (GCE) for simultaneous determination of l-Dopa and benserazide. For the elaboration of the GCE/rGO, the developed methodology was based on the electrochemical technique: multiple pulse amperometry (MPA). The MPA was more stable and efficient for the formation of rGO film, under optimum conditions (pH 6.00; concentration of rGO 2.00 mg mL−1; time 450 s; potentials −0.60, −0.70, −0.80, −0.90, −0.95, −1.00, −1.10, −1.20, and – 1.30 V). After the film was formed, the cyclic voltammetry was used to detect LD and BZ in real samples and optimized conditions 0.05 mol L−1 PBS (pH 5.50). The linear range for the LD is 25–425 μmol L−1 and the BZ of 5–80 μmol L−1. The limit of detection calculated was 17.10 (LD) and 2.99 (BZ) μmol L−1.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/70121",risUrl:"/chapter/ris/70121",signatures:"Thiago Gabry Barbosa, Ana Elisa Ferreira Oliveira and Arnaldo César Pereira",book:{id:"9305",type:"book",title:"Graphene Production and Application",subtitle:null,fullTitle:"Graphene Production and Application",slug:"graphene-production-and-application",publishedDate:"May 6th 2020",bookSignature:"Sadia Ameen, M. Shaheer Akhtar and Hyung-Shik Shin",coverURL:"https://cdn.intechopen.com/books/images_new/9305.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",isbn:"978-1-83880-192-2",printIsbn:"978-1-83880-191-5",pdfIsbn:"978-1-83880-252-3",isAvailableForWebshopOrdering:!0,editors:[{id:"52613",title:"Dr.",name:"Sadia",middleName:null,surname:"Ameen",slug:"sadia-ameen",fullName:"Sadia Ameen"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:[{id:"307892",title:"Prof.",name:"Arnaldo César",middleName:null,surname:"Pereira",fullName:"Arnaldo César Pereira",slug:"arnaldo-cesar-pereira",email:"arnaldocsp@yahoo.com.br",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"307894",title:"Mr.",name:"Thiago Gabry",middleName:null,surname:"Barbosa",fullName:"Thiago Gabry Barbosa",slug:"thiago-gabry-barbosa",email:"thgabry@hotmail.com",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null},{id:"307895",title:"Mrs.",name:"Ana Elisa",middleName:null,surname:"Ferreira Oliveira",fullName:"Ana Elisa Ferreira Oliveira",slug:"ana-elisa-ferreira-oliveira",email:"ana_elisa_oliveira@yahoo.com",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Construction of an electrochemical sensor based on electroreduction of graphene oxide on a GCE",level:"1"},{id:"sec_2_2",title:"2.1 Instrumentation",level:"2"},{id:"sec_3_2",title:"2.2 Pretreatment of glassy carbon electrode",level:"2"},{id:"sec_4_2",title:"2.3 Electroreduction of graphene oxide",level:"2"},{id:"sec_5_2",title:"2.4 Multiple pulse amperometry",level:"2"},{id:"sec_7",title:"3. Simultaneous determination of l-Dopa and benserazide",level:"1"},{id:"sec_7_2",title:"3.1 Electrochemical behavior of l-Dopa and benserazide",level:"2"},{id:"sec_8_2",title:"3.2 Optimization of experimental parameters",level:"2"},{id:"sec_8_3",title:"3.2.1 Effect of applied potential range",level:"3"},{id:"sec_9_3",title:"3.2.2 Effect of time",level:"3"},{id:"sec_10_3",title:"3.2.3 Effect of GO concentration",level:"3"},{id:"sec_11_3",title:"3.2.4 Effect of pH",level:"3"},{id:"sec_13_2",title:"3.3 Optimization of experimental conditions",level:"2"},{id:"sec_13_3",title:"3.3.1 Influence of ionic force",level:"3"},{id:"sec_14_3",title:"3.3.2 pH influence",level:"3"},{id:"sec_16_2",title:"3.4 Calibration curve",level:"2"},{id:"sec_17_2",title:"3.5 Application in pharmaceutical formulation",level:"2"},{id:"sec_19",title:"4. Conclusion",level:"1"},{id:"sec_20",title:"Acknowledgments",level:"1"}],chapterReferences:[{id:"B1",body:'\nDorsey ER, Sherer T, Okun MS, Bloem RB. The emerging evidence of the Parkinson pandemic. Journal of Parkison’s Disease. 2018;8(Suppl 1):S3-S8. DOI: 10.3233/JPD-181474\n'},{id:"B2",body:'\nVenton BJ, Wightman RM. Psychoanalytical electrochemistry: Dopamine and behavior. Analytical Chemistry. 2003;75:414A-421A. DOI: 10.1021/ac031421c\n'},{id:"B3",body:'\nOlanow CW, Obeso JA, Stocchi F. Continuous dopaminereceptor treatment of Parkinson’s disease: Scientific rationale and clinical implications. Lancet Neurology. 2006;5:677-687. DOI: 10.1016/S1474-4422 (06) 70521-X\n'},{id:"B4",body:'\nHershey T, Black KJ, Carl JL, Mcgee-Minnich L, Snyder AZ. Long term treatment and disease severity change brain responses to levodopa in Parkinson\'s disease. Journal of Neurology, Neurosurgery, and Psychiatry. 2003;74:844-851. DOI: 10.1136/jnnp.74.7.844\n'},{id:"B5",body:'\nKhor SP, Hsu A. The pharmacokinetics and pharmacodynamics of levodopa in the treatment of Parkinsons disease. Current Clinical Pharmacology. 2007;2(3):234-243. DOI: 10.2174/157488407781668802\n'},{id:"B6",body:'\nRezaei B, Shams-Ghahfarokhi L, Havakeshian E, Ensafi AA. An electrochemical biosensor based on nanoporous stainless steel modified by gold and palladium nanoparticles for simultaneous determination of levodopa and uric acid. Talanta. 2016;158:42-50. DOI: 10.1016/j.talanta.2016.04.061\n'},{id:"B7",body:'\nBurkhard P, Dominici P, Borri-Voltattorni C, Jansonius JN, Malashkevich VN. Structural insight into Parkinson’s disease treatment from drug-inhibited DOPA decarboxylase. Nature Structural Biology. 2001;8(11):963-967. DOI: 10.1038/nsb1101-963\n'},{id:"B8",body:'\nTreseder SA, Rose S, Summo L, Jenner P. Commonly used Lamino acid decarboxylase inhibitors block monoamine oxidase activity in the rat. Journal of Neural Transmission. 2003;110(3):229-238. DOI: 10.1007/s00702-002-0778-4\n'},{id:"B9",body:'\nNorouzi P, Larijani B, Faridbod F, Ganjali MR. A novel method for ultra trace measurement of bentazon based on nanocomposite electrode and continuous coulometric FFTCyclic voltammetry. International Journal of Environmental Research. 2015;9:101-108\n'},{id:"B10",body:'\nWang Y, Hasebe Y. Uricase-adsorbed carbon-felt reactor coupled with a peroxidase-modified carbon-felt-based H2O2 detector for highly sensitive amperometric flow determination of uric acid. Journal of Pharmaceutical and Biomedical Analysis. 2012;57:125-132. DOI: 10.1016/j.jpba.2011.08.021\n'},{id:"B11",body:'\nTrojanowicz M. Recent developments in electrochemical flow detections—A review: Part II. Liquid chromatography. Analytica Chemica Acta. 2011;688:8-35. DOI: 10.1016/j.aca.2010.12.024\n'},{id:"B12",body:'\nde Miranda JAT. Simultaneous determination of aspirin and ascorbic acid in medications using flow injection analysis with amperometric detection [master’s thesis]. Uberlândia: Federal University of Uberlândia—Graduate Program in Chemistry; 2011. DOI: 10.1002/elan.200804262\n'},{id:"B13",body:'\nPereira AC. Electrochemical behavior of some electroactive organic compounds immobilized in inorganic media aiming at the development of sensors for NADH [PhD thesis]. Unicamp: Institute of Chemistry; 2003\n'},{id:"B14",body:'\nOliveira AEF, Braga GB, Tarley CRT, Pereira AC. Thermally reduced graphene oxide: Synthesis, studies and characterization. Journal of Materials Science. 2018;53:12005-12015. DOI: 10.1007/s10853-018-2473-3\n'},{id:"B15",body:'\nLi Y, Martens I, Cheung KC, Bizzotto D. Electrodeposition of reduced graphene oxide onto gold electrodes: Creating thin electrochemically active and optically transparent overlayers. Electrochimica Acta. 2019;319:649-656. DOI: 10.1016/j.electacta.2019.07.004\n'},{id:"B16",body:'\nYao SY, Cai WL, Liu L, Liao XQ , Tao KL, Feng F, et al. Electrochemical behavior of eriocitrin and highly sensitive determination based on an electrochemically reduced graphene oxide modified glassy carbon electrode. Analytical Methods. 2016;18:3722-3729. DOI: 10.1039/C6AY00064A\n'},{id:"B17",body:'\nWang X, Shen W, Zhang X, Guo S, Gao Y, Li X, et al. Indirect electrochemical determination of ribavirin using boronic acid-diol recognition on a 3-aminophenylboronic acid-electrochemically reduced graphene oxide modified glassy carbon electrode (APBA/ERGO/GCE). Analytical Letters. 2019;52:1-14. DOI: 10.1080/00032719.2019.1576716\n'},{id:"B18",body:'\nMotta R, Schrebler R, Zanoni R, Dalchiele EA. Insights from experiment and theory into the electrochemical reduction mechanism of graphene oxide. Electrochimica Acta. 2019;304:231-238. DOI: 10.1016/j.electacta.2019.02.108\n'},{id:"B19",body:'\nde Camargo MNL et al. Tuning the electrochemical reduction of graphene oxide: Structural correlations towards the electrooxidation of nicotinamide adenine dinucleotide hydride. Electrochimica Acta. 2016;197:194-199. DOI: 10.1016/j.electacta.2015.09.022\n'},{id:"B20",body:'\nKumar DR, Kesavan S, Bayonosa ML, Shin JJ. 3,5-Diamino-1,2,4-triazole@electrochemically reduced graphene oxide film modified electrode for the electrochemical determination of 4-nitrophenol. Electrochimica Acta. 2017;246:1131-1140. DOI: 10.1016/j.electacta.2017.06.116\n'},{id:"B21",body:'\nPalakollu VN et al. Electrochemically reduced graphene oxide/poly-glycine composite modified electrode for sensitive determination of l-dopa. Materials Science and Engineering: C. 2017;77:394-404. DOI: 10.1016/j.msec.2017.03.173\n'},{id:"B22",body:'\nGarcía-Argumáneza A, Llorentea I, Caballero-Calerob O, Gonzálezc Z, Menéndezc R, Escuderoa ML, et al. Electrochemical reduction of graphene oxide on biomedical grade CoCr alloy. Applied Surface Science. 2019;465:1028-1036. DOI: 10.1016/j.apsusc.2018.09.188\n'},{id:"B23",body:'\nSunder GSS, Rohanifar A, Devasurendra AM, Kirchhoff JR. Selective determination of l-DOPA at a graphene oxide/yttrium oxide modified glassy carbon electrode. Electrochimica Acta. 2019;301:192-199. DOI: 10.1016/j.electacta.2019.01.098\n'},{id:"B24",body:'\nHassanvand Z, Jalali F. Simultaneous determination of l-DOPA, l-tyrosine and uric acid by cysteic acid-modified glassy carbon electrode. Materials Science and Engineering C. 2019;98:496-502. DOI: 10.1016/j.msec.2018.12.131\n'},{id:"B25",body:'\nPrabhu P, Babu RS, Narayanan SS. Amperometric determination of l-dopa by nickel hexacyanoferrate film modified gold nanoparticle graphite composite electrode. Sensors and Actuators B: Chemical. 2011;156(2):606-614. DOI: 10.1016/j.snb.2011.02.006\n'},{id:"B26",body:'\nSufredini HB, Pedrosa VA, Codognoto L, Machado SAS, Filho RCR, Avaca LA. Enhanced electrochemical response of boron-doped diamond electrodes brought on by a cathodic surface pre-treatment. Electrochimica Acta. 2013;49:4021-4026\n'},{id:"B27",body:'\nNaushad M, Gupta VK, Wabaidur SM, Alothman ZA. Simultaneous determination of benserazide and levodopa in pharmaceutical tablet, human serum and urine sample by differential pulse voltammetry using modified glassy carbon electrode. International Journal of Electrochemical Science. 2013;8:297-311\n'},{id:"B28",body:'\nKul D, Brett CMA. Electrochemical investigation and determination of levodopa on poly(Nile blue-a)/multiwalled carbon nanotube modified glassy carbon electrodes. Electroanalysis. 2014;26:1320-1325. DOI: 10.1002/elan.201400071\n'},{id:"B29",body:'\nZhou YZ, Alany RG, Chuang V, Wen J. Studies of the rate constant of l-DOPA oxidation and decarboxylation by HPLC. Chromatographia. 2012;75:597-606. DOI: 10.1007/s10337-012-2229-1\n'},{id:"B30",body:'\nEnsafi AA, Bahmari H, Rezaei B, Maleh KH. Application of ionic liquid–TiO2 nanoparticle modified carbon paste electrode for the voltammetric determination of benserazide in biological samples. Materials Science and Engineering. 2013;33(2):831-835. DOI: 10.1016/j.msec.2012.11.008\n'},{id:"B31",body:'\nFouladgar M. A high sensitive square wave voltammetric sensor based on ZnO nanoparticle ionic liquid paste electrode for determination of benserazide in biological samples. Measurement. 2016:141-147. DOI: 10.1016/j.measurement.2016.02.057\n'},{id:"B32",body:'\nRabinca AA et al. Voltammetric method for simultaneous determination of l-Dopa and benserazide. Current Analytical Chemistry. 2017;13:218-224. DOI: 10.2174/1573411012666160601161703\n'},{id:"B33",body:'\nRahmanifar E, Yoosefian M, Maleh KH. Application of CdO/SWCNTs nanocomposite ionic liquids carbon paste electrode as a voltammetric sensor for determination of benserazide. Current Analytical Chemistry. 2017;13:46-51. DOI: 10.2174/1573411012666160601145809\n'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"Thiago Gabry Barbosa",address:null,affiliation:'
Departamento de Ciências Naturais, Universidade Federal de São João del-Rei, UFSJ, São João del-Rei, MG, Brazil
Departamento de Ciências Naturais, Universidade Federal de São João del-Rei, UFSJ, São João del-Rei, MG, Brazil
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The increasing technological complexity of these systems is posing serious challenges regarding electromagnetic compatibility (EMC) issues. Indeed, the radiation emitted from electronic circuits can induce harmful effects on nearby devices. Thus, several research works have been conducted using the nearfield technique to deal with electromagnetic interferences (EMI) that might occur, especially due to rapidly changing currents and voltages. In the present work, a detailed study about the characterization of the electromagnetic nearfield-radiated emissions is established using a time-domain analysis to provide an equivalent model constituted of a set of electromagnetic dipole parameters. Source reconstruction has been obtained using electromagnetic time reversal (EMTR), which has proven successful and efficient in identifying transient disturbance sources in power electronics. Experimental measurements of the magnetic nearfield have been carried out under an AC/DC flyback converter. 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The results of a reasonable computing time have shown that, particularly in transient signals with a wide frequency band, the suggested inverse method is an adequate alternative to overcome frequency domain limitations.",signatures:"Sassia Hedia, Bessem Zitouna, Jaleleddine Ben Hadj Slama and Lionel Pichon",authors:[{id:"309925",title:"Ph.D. Student",name:"Sassia",surname:"Hedia",fullName:"Sassia Hedia",slug:"sassia-hedia",email:"hdia.sassia@gmail.com"},{id:"310034",title:"Dr.",name:"Bessem",surname:"Zitouna",fullName:"Bessem Zitouna",slug:"bessem-zitouna",email:"bessem.zitouna@yahoo.fr"},{id:"435309",title:"Prof.",name:"Lionel",surname:"Pichon",fullName:"Lionel Pichon",slug:"lionel-pichon",email:"lionel.pichon@geeps.centralesupelec.fr"}],book:{id:"10852",title:"Recent Topics in Electromagnetic Compatibility",slug:"recent-topics-in-electromagnetic-compatibility",productType:{id:"1",title:"Edited Volume"}}}],collaborators:[{id:"310034",title:"Dr.",name:"Bessem",surname:"Zitouna",slug:"bessem-zitouna",fullName:"Bessem Zitouna",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"310038",title:"Prof.",name:"Jaleleddine",surname:"Ben Hadj Slama",slug:"jaleleddine-ben-hadj-slama",fullName:"Jaleleddine Ben Hadj Slama",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"353926",title:"Dr.",name:"Ting-Wei",surname:"Wang",slug:"ting-wei-wang",fullName:"Ting-Wei Wang",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/353926/images/17405_n.jpg",biography:"Dr. Ting-Wei Wang received the B.S. degree from the Department of Electrical Engineering, National Taiwan University of Science and Technology, Taipei, Taiwan, in 2014, the M.S. degree from the Institute of Electro-Optical Engineering, National Chiao Tung University, Hsinchu, Taiwan, in 2016, and the Ph.D. degree from the Department of Electrical and Computer Engineering, National Chiao Tung University, in 2020. From 2016 to 2019, he was a Research and Development Engineer of advanced process development with Taiwan Semiconductor Manufacturing Company (TSMC), Hsinchu, Taiwan. He is currently a Postdoctoral Researcher with Department of Electrical Engineering and Department of Medical Engineering, California Institute of Technology, Pasadena, CA, USA. 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This study provides, on the other hand, a global vision of the family starting for their fortuitous discovery, the synthesis of their derivatives, their mechanism of action widely known nowadays, the actual classification according to the chemical structure and pharmacokinetic properties, and their uses and indications, the traditional and the new ones. On the other hand,the study is focused in the mainly problems of benzodiazepines, depedence, and tolerance, many times led by a misuse of the patient, wrong prescriptions, or extended treatments. A withdrawal program is proposed that includes the important factors or criteria to success, with a slow and gradual reduction of these drugs, avoiding relapse or severe adverse effects. New lines of research related to benzodiazepines are taken into account, which not only include the new therapeutic uses but also the adverse effects in short and long term. They are also analyzed the new discoveries concerning the nonbenzodiazepine drugs due to the close relation they have with benzodiazepines.",book:{id:"7518",slug:"medicinal-chemistry",title:"Medicinal Chemistry",fullTitle:"Medicinal Chemistry"},signatures:"Elisabet Batlle, Enric Lizano, Miquel Viñas and Maria Dolors Pujol",authors:[{id:"252409",title:"Prof.",name:"Maria Dolors",middleName:null,surname:"Pujol",slug:"maria-dolors-pujol",fullName:"Maria Dolors Pujol"},{id:"262535",title:"Dr.",name:"Elisabet",middleName:null,surname:"Batlle",slug:"elisabet-batlle",fullName:"Elisabet Batlle"},{id:"262756",title:"Mr.",name:"Enric",middleName:null,surname:"Lizano",slug:"enric-lizano",fullName:"Enric Lizano"},{id:"274188",title:"Dr.",name:"Miquel",middleName:null,surname:"Viñas",slug:"miquel-vinas",fullName:"Miquel Viñas"}]},{id:"62647",title:"Indomethacin from Anti-Inflammatory to Anticancer Agent",slug:"indomethacin-from-anti-inflammatory-to-anticancer-agent",totalDownloads:1313,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"The chapter “Indomethacin from Anti-inflammatory to Anticancer Agent” covers the recent reports regarding the implication of COX-2/PGE2 in multiple cancer cell proliferation to emphasize the anticancer potential of COX-inhibitors including indomethacin and to reveal that the reduction of PGE2 production interferes with the cancer cell proliferation belongs to multiple cancer cell types. Impressively, indomethacin is involved in antiproliferative and apoptotic actions against cancer cell types via COX-2-independent mechanisms to highlight indomethacin as promising anticancer agent with dual actions to control the cancer cell proliferation. The cardiovascular complications result from diaryl heterocycle sulfonamide/methylsulfone selective COX-2 inhibitors upon reduction in PGE2 and PGI2 production that affects the vascular tone limits the use of Celecoxib as chemopreventive agent against recurrence of colorectal carcinoma cells. Kinetic profile of indomethacin against COX-2 showed obvious difference from that of selective COX-2 inhibitors in which it recovered completely from the enzyme after long time of incubation while COX-2 inhibitors did not recover to impress that this might be implicated in the cardiovascular toxicity of the selective inhibitors. This raised the concern to develop the indomethacin from nonselective COX- to selective COX-2-inhibitors and to assert whether the cardiac complications are from pharmacological class effect or chemical class effect.",book:{id:"7518",slug:"medicinal-chemistry",title:"Medicinal Chemistry",fullTitle:"Medicinal Chemistry"},signatures:"Shaymaa Emam Kassab",authors:[{id:"251335",title:"Dr.",name:"Shaymaa",middleName:null,surname:"Kassab",slug:"shaymaa-kassab",fullName:"Shaymaa Kassab"}]},{id:"63789",title:"Clinical Pharmacokinetics of Clavulanic Acid, a Novel β- Lactamase Isolated from Streptomyces clavuligerus and Its Variability",slug:"clinical-pharmacokinetics-of-clavulanic-acid-a-novel-lactamase-isolated-from-streptomyces-clavuliger",totalDownloads:1099,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"The clavulanic acid derived by fermentation of Streptomyces clavuligerus and possessed the capability to inactivate a broad range of β-lactamase enzymes. A complex physicochemical process involves the binding of clavulanic acid to β-lactamases in which clavulanic acid itself deplete irreversibly along with β-lactamase enzyme rendering amoxicillin spared which otherwise would hydrolyze by an enzyme. It is therefore termed as ‘suicide ‘inhibitor for β-lactamases. We discussed here pharmacokinetic parameters and identified factors responsible for the variability of absorption of clavulanic acid. The results based on individual plasma concentration-time curve of amoxicillin and clavulanic acid in an open, randomized, two-way crossover study involving 10 healthy male subjects administered with two amoxiclav formulations.",book:{id:"7518",slug:"medicinal-chemistry",title:"Medicinal Chemistry",fullTitle:"Medicinal Chemistry"},signatures:"Anab Fatima, Mohammad Jiyad Shaikh, Hina Zahid, Ishart Younus,\nSheikh Abdul Khaliq and Farah Khalid",authors:[{id:"225358",title:"Dr.",name:"Muhammad Jiyad",middleName:null,surname:"Shaikh",slug:"muhammad-jiyad-shaikh",fullName:"Muhammad Jiyad Shaikh"},{id:"231412",title:"Dr.",name:"Anab",middleName:null,surname:"Fatima",slug:"anab-fatima",fullName:"Anab Fatima"},{id:"243371",title:"Dr.",name:"Hina",middleName:null,surname:"Zahid",slug:"hina-zahid",fullName:"Hina Zahid"},{id:"243372",title:"Dr.",name:"Ishart",middleName:null,surname:"Younus",slug:"ishart-younus",fullName:"Ishart Younus"},{id:"243373",title:"Dr.",name:"Sheikh Abdul",middleName:null,surname:"Khaliq",slug:"sheikh-abdul-khaliq",fullName:"Sheikh Abdul Khaliq"},{id:"243374",title:"Dr.",name:"Farah",middleName:null,surname:"Khalid",slug:"farah-khalid",fullName:"Farah Khalid"}]},{id:"63353",title:"New Antituberculosis Drug FS-1",slug:"new-antituberculosis-drug-fs-1",totalDownloads:1460,totalCrossrefCites:2,totalDimensionsCites:5,abstract:"The new iodine complex (FS-1), including molecular iodine, which is coordinated by lithium, magnesium halides, and bioorganic ligands, possesses high bactericidal activity against various microorganisms, including Mycobacterium sp., Staphylococcus aureus MRSA and MSSA, Escherichia coli, Pseudomonas aeruginosa, etc. FS-1 has synergistic properties with a broad class of antibiotics. The experimental model of tuberculosis in guinea pigs caused by clinical multidrug-resistant strains of Mycobacterium tuberculosis shows antituberculosis, immunomodulatory, and anti-inflammatory activity. FS-1 is characterized by low acute toxicity and lack of genotoxicity and mutagenicity. FS-1 is well distributed to organs and tissues; its pharmacokinetics is linear. The maximum nontoxic dose is 100 mg/kg for rats after 28-day oral administration and 30 mg/kg for rabbits after 180-day oral administration.",book:{id:"7518",slug:"medicinal-chemistry",title:"Medicinal Chemistry",fullTitle:"Medicinal Chemistry"},signatures:"Rinat Islamov, Bahkytzhan Kerimzhanova and Alexander Ilin",authors:[{id:"109493",title:"Dr.",name:"Rinat",middleName:null,surname:"Islamov",slug:"rinat-islamov",fullName:"Rinat Islamov"},{id:"136527",title:"Dr.",name:"Alexander",middleName:null,surname:"Ilin",slug:"alexander-ilin",fullName:"Alexander Ilin"},{id:"261645",title:"Prof.",name:"Bahyitzhan",middleName:null,surname:"Kerimzhanova",slug:"bahyitzhan-kerimzhanova",fullName:"Bahyitzhan Kerimzhanova"}]},{id:"64761",title:"Introductory Chapter: Unregulated Mitochondrial Production of Reactive Oxygen Species in Testing the Biological Activity of Compounds",slug:"introductory-chapter-unregulated-mitochondrial-production-of-reactive-oxygen-species-in-testing-the-",totalDownloads:974,totalCrossrefCites:0,totalDimensionsCites:1,abstract:null,book:{id:"7518",slug:"medicinal-chemistry",title:"Medicinal Chemistry",fullTitle:"Medicinal Chemistry"},signatures:"Janka Vašková and Ladislav Vaško",authors:[{id:"140747",title:"Associate Prof.",name:"Janka",middleName:null,surname:"Vašková",slug:"janka-vaskova",fullName:"Janka Vašková"},{id:"207199",title:"Prof.",name:"Ladislav",middleName:null,surname:"Vaško",slug:"ladislav-vasko",fullName:"Ladislav Vaško"}]}],onlineFirstChaptersFilter:{topicId:"1190",limit:6,offset:0},onlineFirstChaptersCollection:[],onlineFirstChaptersTotal:0},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:0,limit:8,total:null},allSeries:{pteSeriesList:[],lsSeriesList:[],hsSeriesList:[],sshSeriesList:[],testimonialsList:[]},series:{item:{id:"11",title:"Biochemistry",doi:"10.5772/intechopen.72877",issn:"2632-0983",scope:"Biochemistry, the study of chemical transformations occurring within living organisms, impacts all areas of life sciences, from molecular crystallography and genetics to ecology, medicine, and population biology. Biochemistry examines macromolecules - proteins, nucleic acids, carbohydrates, and lipids – and their building blocks, structures, functions, and interactions. 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. This Biochemistry Series will address the current research on biomolecules and the emerging trends with great promise.",coverUrl:"https://cdn.intechopen.com/series/covers/11.jpg",latestPublicationDate:"May 18th, 2022",hasOnlineFirst:!0,numberOfPublishedBooks:27,editor:{id:"31610",title:"Dr.",name:"Miroslav",middleName:null,surname:"Blumenberg",slug:"miroslav-blumenberg",fullName:"Miroslav Blumenberg",profilePictureURL:"https://mts.intechopen.com/storage/users/31610/images/system/31610.jpg",biography:"Miroslav Blumenberg, Ph.D., was born in Subotica and received his BSc in Belgrade, Yugoslavia. He completed his Ph.D. at MIT in Organic Chemistry; he followed up his Ph.D. with two postdoctoral study periods at Stanford University. Since 1983, he has been a faculty member of the RO Perelman Department of Dermatology, NYU School of Medicine, where he is codirector of a training grant in cutaneous biology. 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. He is a Consultant Reviewer for several journals, including the Journal of Chromatography A, Journal of Chromatography B, Plos ONE, Proteomes, International Journal of Molecular Science, Biotech, Electrophoresis, and others. He is also Associate Editor of Biotech.",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorTwo:{id:"201414",title:"Dr.",name:"Simona",middleName:null,surname:"Viglio",slug:"simona-viglio",fullName:"Simona Viglio",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRKDHQA4/Profile_Picture_1630402531487",biography:"Simona Viglio is an Associate Professor of Biochemistry at the Department of Molecular Medicine at the University of Pavia. She has been working since 1995 on the determination of proteolytic enzymes involved in the degradation process of connective tissue matrix and on the identification of biological markers of lung diseases. 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. She is an author of about 90 publications (According to Scopus: H-Index: 23; According to WOS: H-Index: 20) on peer-reviewed journals, a member of the “Società Italiana di Biochimica e Biologia Molecolare,“ and a Consultant Reviewer for International Journal of Molecular Science, Journal of Chromatography A, COPD, Plos ONE and Nutritional Neuroscience.",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorThree:null}]},overviewPageOFChapters:{paginationCount:49,paginationItems:[{id:"80495",title:"Iron in Cell Metabolism and Disease",doi:"10.5772/intechopen.101908",signatures:"Eeka Prabhakar",slug:"iron-in-cell-metabolism-and-disease",totalDownloads:1,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Iron Metabolism - Iron a Double‐Edged Sword",coverURL:"https://cdn.intechopen.com/books/images_new/10842.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"81799",title:"Cross Talk of Purinergic and Immune Signaling: Implication in Inflammatory and Pathogenic Diseases",doi:"10.5772/intechopen.104978",signatures:"Richa Rai",slug:"cross-talk-of-purinergic-and-immune-signaling-implication-in-inflammatory-and-pathogenic-diseases",totalDownloads:7,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Purinergic System",coverURL:"https://cdn.intechopen.com/books/images_new/10801.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"81764",title:"Involvement of the Purinergic System in Cell Death in Models of Retinopathies",doi:"10.5772/intechopen.103935",signatures:"Douglas Penaforte Cruz, Marinna Garcia Repossi and Lucianne Fragel Madeira",slug:"involvement-of-the-purinergic-system-in-cell-death-in-models-of-retinopathies",totalDownloads:4,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Purinergic System",coverURL:"https://cdn.intechopen.com/books/images_new/10801.jpg",subseries:{id:"17",title:"Metabolism"}}},{id:"81756",title:"Alteration of Cytokines Level and Oxidative Stress Parameters in COVID-19",doi:"10.5772/intechopen.104950",signatures:"Marija Petrusevska, Emilija Atanasovska, Dragica Zendelovska, Aleksandar Eftimov and Katerina Spasovska",slug:"alteration-of-cytokines-level-and-oxidative-stress-parameters-in-covid-19",totalDownloads:8,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Chemokines Updates",coverURL:"https://cdn.intechopen.com/books/images_new/11672.jpg",subseries:{id:"18",title:"Proteomics"}}}]},overviewPagePublishedBooks:{paginationCount:27,paginationItems:[{type:"book",id:"7006",title:"Biochemistry and Health Benefits of Fatty Acids",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/7006.jpg",slug:"biochemistry-and-health-benefits-of-fatty-acids",publishedDate:"December 19th 2018",editedByType:"Edited by",bookSignature:"Viduranga Waisundara",hash:"c93a00abd68b5eba67e5e719f67fd20b",volumeInSeries:1,fullTitle:"Biochemistry and Health Benefits of Fatty Acids",editors:[{id:"194281",title:"Dr.",name:"Viduranga Y.",middleName:null,surname:"Waisundara",slug:"viduranga-y.-waisundara",fullName:"Viduranga Y. Waisundara",profilePictureURL:"https://mts.intechopen.com/storage/users/194281/images/system/194281.jpg",biography:"Dr. Viduranga Waisundara obtained her Ph.D. in Food Science and Technology from the Department of Chemistry, National University of Singapore, in 2010. She was a lecturer at Temasek Polytechnic, Singapore from July 2009 to March 2013. She relocated to her motherland of Sri Lanka and spearheaded the Functional Food Product Development Project at the National Institute of Fundamental Studies from April 2013 to October 2016. She was a senior lecturer on a temporary basis at the Department of Food Technology, Faculty of Technology, Rajarata University of Sri Lanka. She is currently Deputy Principal of the Australian College of Business and Technology – Kandy Campus, Sri Lanka. She is also the Global Harmonization Initiative (GHI) Ambassador to Sri Lanka.",institutionString:"Australian College of Business & Technology",institution:null}]},{type:"book",id:"6820",title:"Keratin",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/6820.jpg",slug:"keratin",publishedDate:"December 19th 2018",editedByType:"Edited by",bookSignature:"Miroslav Blumenberg",hash:"6def75cd4b6b5324a02b6dc0359896d0",volumeInSeries:2,fullTitle:"Keratin",editors:[{id:"31610",title:"Dr.",name:"Miroslav",middleName:null,surname:"Blumenberg",slug:"miroslav-blumenberg",fullName:"Miroslav Blumenberg",profilePictureURL:"https://mts.intechopen.com/storage/users/31610/images/system/31610.jpg",biography:"Miroslav Blumenberg, Ph.D., was born in Subotica and received his BSc in Belgrade, Yugoslavia. He completed his Ph.D. at MIT in Organic Chemistry; he followed up his Ph.D. with two postdoctoral study periods at Stanford University. Since 1983, he has been a faculty member of the RO Perelman Department of Dermatology, NYU School of Medicine, where he is codirector of a training grant in cutaneous biology. 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"}}}]},{type:"book",id:"7978",title:"Vitamin A",subtitle:null,coverURL:"https://cdn.intechopen.com/books/images_new/7978.jpg",slug:"vitamin-a",publishedDate:"May 15th 2019",editedByType:"Edited by",bookSignature:"Leila Queiroz Zepka, Veridiana Vera de Rosso and Eduardo Jacob-Lopes",hash:"dad04a658ab9e3d851d23705980a688b",volumeInSeries:3,fullTitle:"Vitamin A",editors:[{id:"261969",title:"Dr.",name:"Leila",middleName:null,surname:"Queiroz Zepka",slug:"leila-queiroz-zepka",fullName:"Leila Queiroz Zepka",profilePictureURL:"https://mts.intechopen.com/storage/users/261969/images/system/261969.png",biography:"Prof. Dr. Leila Queiroz Zepka is currently an associate professor in the Department of Food Technology and Science, Federal University of Santa Maria, Brazil. She has more than fifteen years of teaching and research experience. She has published more than 550 scientific publications/communications, including 15 books, 50 book chapters, 100 original research papers, 380 research communications in national and international conferences, and 12 patents. She is a member of the editorial board of five journals and acts as a reviewer for several national and international journals. Her research interests include microalgal biotechnology with an emphasis on microalgae-based products.",institutionString:"Universidade Federal de Santa Maria",institution:{name:"Universidade Federal de Santa Maria",institutionURL:null,country:{name:"Brazil"}}}]},{type:"book",id:"7953",title:"Bioluminescence",subtitle:"Analytical Applications and Basic Biology",coverURL:"https://cdn.intechopen.com/books/images_new/7953.jpg",slug:"bioluminescence-analytical-applications-and-basic-biology",publishedDate:"September 25th 2019",editedByType:"Edited by",bookSignature:"Hirobumi Suzuki",hash:"3a8efa00b71abea11bf01973dc589979",volumeInSeries:4,fullTitle:"Bioluminescence - Analytical Applications and Basic Biology",editors:[{id:"185746",title:"Dr.",name:"Hirobumi",middleName:null,surname:"Suzuki",slug:"hirobumi-suzuki",fullName:"Hirobumi Suzuki",profilePictureURL:"https://mts.intechopen.com/storage/users/185746/images/system/185746.png",biography:"Dr. Hirobumi Suzuki received his Ph.D. in 1997 from Tokyo Metropolitan University, Japan, where he studied firefly phylogeny and the evolution of mating systems. He is especially interested in the genetic differentiation pattern and speciation process that correlate to the flashing pattern and mating behavior of some fireflies in Japan. He then worked for Olympus Corporation, a Japanese manufacturer of optics and imaging products, where he was involved in the development of luminescence technology and produced a bioluminescence microscope that is currently being used for gene expression analysis in chronobiology, neurobiology, and developmental biology. Dr. Suzuki currently serves as a visiting researcher at Kogakuin University, Japan, and also a vice president of the Japan Firefly Society.",institutionString:"Kogakuin University",institution:null}]}]},openForSubmissionBooks:{},onlineFirstChapters:{},subseriesFiltersForOFChapters:[],publishedBooks:{},subseriesFiltersForPublishedBooks:[],publicationYearFilters:[],authors:{paginationCount:148,paginationItems:[{id:"165328",title:"Dr.",name:"Vahid",middleName:null,surname:"Asadpour",slug:"vahid-asadpour",fullName:"Vahid Asadpour",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/165328/images/system/165328.jpg",biography:"Vahid Asadpour, MS, Ph.D., is currently with the Department of Research and Evaluation, Kaiser Permanente Southern California. He has both an MS and Ph.D. in Biomedical Engineering. He was previously a research scientist at the University of California Los Angeles (UCLA) and visiting professor and researcher at the University of North Dakota. He is currently working in artificial intelligence and its applications in medical signal processing. In addition, he is using digital signal processing in medical imaging and speech processing. Dr. Asadpour has developed brain-computer interfacing algorithms and has published books, book chapters, and several journal and conference papers in this field and other areas of intelligent signal processing. He has also designed medical devices, including a laser Doppler monitoring system.",institutionString:"Kaiser Permanente Southern California",institution:null},{id:"169608",title:"Prof.",name:"Marian",middleName:null,surname:"Găiceanu",slug:"marian-gaiceanu",fullName:"Marian Găiceanu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/169608/images/system/169608.png",biography:"Prof. Dr. Marian Gaiceanu graduated from the Naval and Electrical Engineering Faculty, Dunarea de Jos University of Galati, Romania, in 1997. He received a Ph.D. (Magna Cum Laude) in Electrical Engineering in 2002. Since 2017, Dr. Gaiceanu has been a Ph.D. supervisor for students in Electrical Engineering. He has been employed at Dunarea de Jos University of Galati since 1996, where he is currently a professor. Dr. Gaiceanu is a member of the National Council for Attesting Titles, Diplomas and Certificates, an expert of the Executive Agency for Higher Education, Research Funding, and a member of the Senate of the Dunarea de Jos University of Galati. He has been the head of the Integrated Energy Conversion Systems and Advanced Control of Complex Processes Research Center, Romania, since 2016. He has conducted several projects in power converter systems for electrical drives, power quality, PEM and SOFC fuel cell power converters for utilities, electric vehicles, and marine applications with the Department of Regulation and Control, SIEI S.pA. (2002–2004) and the Polytechnic University of Turin, Italy (2002–2004, 2006–2007). He is a member of the Institute of Electrical and Electronics Engineers (IEEE) and cofounder-member of the IEEE Power Electronics Romanian Chapter. He is a guest editor at Energies and an academic book editor for IntechOpen. He is also a member of the editorial boards of the Journal of Electrical Engineering, Electronics, Control and Computer Science and Sustainability. Dr. Gaiceanu has been General Chairman of the IEEE International Symposium on Electrical and Electronics Engineering in the last six editions.",institutionString:'"Dunarea de Jos" University of Galati',institution:{name:'"Dunarea de Jos" University of Galati',country:{name:"Romania"}}},{id:"4519",title:"Prof.",name:"Jaydip",middleName:null,surname:"Sen",slug:"jaydip-sen",fullName:"Jaydip Sen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/4519/images/system/4519.jpeg",biography:"Jaydip Sen is associated with Praxis Business School, Kolkata, India, as a professor in the Department of Data Science. His research areas include security and privacy issues in computing and communication, intrusion detection systems, machine learning, deep learning, and artificial intelligence in the financial domain. He has more than 200 publications in reputed international journals, refereed conference proceedings, and 20 book chapters in books published by internationally renowned publishing houses, such as Springer, CRC press, IGI Global, etc. Currently, he is serving on the editorial board of the prestigious journal Frontiers in Communications and Networks and in the technical program committees of a number of high-ranked international conferences organized by the IEEE, USA, and the ACM, USA. He has been listed among the top 2% of scientists in the world for the last three consecutive years, 2019 to 2021 as per studies conducted by the Stanford University, USA.",institutionString:"Praxis Business School",institution:null},{id:"320071",title:"Dr.",name:"Sidra",middleName:null,surname:"Mehtab",slug:"sidra-mehtab",fullName:"Sidra Mehtab",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00002v6KHoQAM/Profile_Picture_1584512086360",biography:"Sidra Mehtab has completed her BS with honors in Physics from Calcutta University, India in 2018. She has done MS in Data Science and Analytics from Maulana Abul Kalam Azad University of Technology (MAKAUT), Kolkata, India in 2020. Her research areas include Econometrics, Time Series Analysis, Machine Learning, Deep Learning, Artificial Intelligence, and Computer and Network Security with a particular focus on Cyber Security Analytics. Ms. Mehtab has published seven papers in international conferences and one of her papers has been accepted for publication in a reputable international journal. She has won the best paper awards in two prestigious international conferences – BAICONF 2019, and ICADCML 2021, organized in the Indian Institute of Management, Bangalore, India in December 2019, and SOA University, Bhubaneswar, India in January 2021. Besides, Ms. Mehtab has also published two book chapters in two books. Seven of her book chapters will be published in a volume shortly in 2021 by Cambridge Scholars’ Press, UK. Currently, she is working as the joint editor of two edited volumes on Time Series Analysis and Forecasting to be published in the first half of 2021 by an international house. Currently, she is working as a Data Scientist with an MNC in Delhi, India.",institutionString:"NSHM College of Management and Technology",institution:null},{id:"226240",title:"Dr.",name:"Andri Irfan",middleName:null,surname:"Rifai",slug:"andri-irfan-rifai",fullName:"Andri Irfan Rifai",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/226240/images/7412_n.jpg",biography:"Andri IRFAN is a Senior Lecturer of Civil Engineering and Planning. He completed the PhD at the Universitas Indonesia & Universidade do Minho with Sandwich Program Scholarship from the Directorate General of Higher Education and LPDP scholarship. He has been teaching for more than 19 years and much active to applied his knowledge in the project construction in Indonesia. His research interest ranges from pavement management system to advanced data mining techniques for transportation engineering. He has published more than 50 papers in journals and 2 books.",institutionString:null,institution:{name:"Universitas Internasional Batam",country:{name:"Indonesia"}}},{id:"314576",title:"Dr.",name:"Ibai",middleName:null,surname:"Laña",slug:"ibai-lana",fullName:"Ibai Laña",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/314576/images/system/314576.jpg",biography:"Dr. Ibai Laña works at TECNALIA as a data analyst. He received his Ph.D. in Artificial Intelligence from the University of the Basque Country (UPV/EHU), Spain, in 2018. He is currently a senior researcher at TECNALIA. His research interests fall within the intersection of intelligent transportation systems, machine learning, traffic data analysis, and data science. He has dealt with urban traffic forecasting problems, applying machine learning models and evolutionary algorithms. He has experience in origin-destination matrix estimation or point of interest and trajectory detection. Working with large volumes of data has given him a good command of big data processing tools and NoSQL databases. He has also been a visiting scholar at the Knowledge Engineering and Discovery Research Institute, Auckland University of Technology.",institutionString:"TECNALIA Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"314575",title:"Dr.",name:"Jesus",middleName:null,surname:"L. Lobo",slug:"jesus-l.-lobo",fullName:"Jesus L. Lobo",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/314575/images/system/314575.png",biography:"Dr. Jesús López is currently based in Bilbao (Spain) working at TECNALIA as Artificial Intelligence Research Scientist. In most cases, a project idea or a new research line needs to be investigated to see if it is good enough to take into production or to focus on it. That is exactly what he does, diving into Machine Learning algorithms and technologies to help TECNALIA to decide whether something is great in theory or will actually impact on the product or processes of its projects. So, he is expert at framing experiments, developing hypotheses, and proving whether they’re true or not, in order to investigate fundamental problems with a longer time horizon. He is also able to design and develop PoCs and system prototypes in simulation. He has participated in several national and internacional R&D projects.\n\nAs another relevant part of his everyday research work, he usually publishes his findings in reputed scientific refereed journals and international conferences, occasionally acting as reviewer and Programme Commitee member. Concretely, since 2018 he has published 9 JCR (8 Q1) journal papers, 9 conference papers (e.g. ECML PKDD 2021), and he has co-edited a book. He is also active in popular science writing data science stories for reputed blogs (KDNuggets, TowardsDataScience, Naukas). Besides, he has recently embarked on mentoring programmes as mentor, and has also worked as data science trainer.",institutionString:"TECNALIA Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"103779",title:"Prof.",name:"Yalcin",middleName:null,surname:"Isler",slug:"yalcin-isler",fullName:"Yalcin Isler",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRyQ8QAK/Profile_Picture_1628834958734",biography:"Yalcin Isler (1971 - Burdur / Turkey) received the B.Sc. degree in the Department of Electrical and Electronics Engineering from Anadolu University, Eskisehir, Turkey, in 1993, the M.Sc. degree from the Department of Electronics and Communication Engineering, Suleyman Demirel University, Isparta, Turkey, in 1996, the Ph.D. degree from the Department of Electrical and Electronics Engineering, Dokuz Eylul University, Izmir, Turkey, in 2009, and the Competence of Associate Professorship from the Turkish Interuniversity Council in 2019.\n\nHe was Lecturer at Burdur Vocational School in Suleyman Demirel University (1993-2000, Burdur / Turkey), Software Engineer (2000-2002, Izmir / Turkey), Research Assistant in Bulent Ecevit University (2002-2003, Zonguldak / Turkey), Research Assistant in Dokuz Eylul University (2003-2010, Izmir / Turkey), Assistant Professor at the Department of Electrical and Electronics Engineering in Bulent Ecevit University (2010-2012, Zonguldak / Turkey), Assistant Professor at the Department of Biomedical Engineering in Izmir Katip Celebi University (2012-2019, Izmir / Turkey). He is an Associate Professor at the Department of Biomedical Engineering at Izmir Katip Celebi University, Izmir / Turkey, since 2019. In addition to academics, he has also founded Islerya Medical and Information Technologies Company, Izmir / Turkey, since 2017.\n\nHis main research interests cover biomedical signal processing, pattern recognition, medical device design, programming, and embedded systems. He has many scientific papers and participated in several projects in these study fields. He was an IEEE Student Member (2009-2011) and IEEE Member (2011-2014) and has been IEEE Senior Member since 2014.",institutionString:null,institution:{name:"Izmir Kâtip Çelebi University",country:{name:"Turkey"}}},{id:"339677",title:"Dr.",name:"Mrinmoy",middleName:null,surname:"Roy",slug:"mrinmoy-roy",fullName:"Mrinmoy Roy",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/339677/images/16768_n.jpg",biography:"An accomplished Sales & Marketing professional with 12 years of cross-functional experience in well-known organisations such as CIPLA, LUPIN, GLENMARK, ASTRAZENECA across different segment of Sales & Marketing, International Business, Institutional Business, Product Management, Strategic Marketing of HIV, Oncology, Derma, Respiratory, Anti-Diabetic, Nutraceutical & Stomatological Product Portfolio and Generic as well as Chronic Critical Care Portfolio. A First Class MBA in International Business & Strategic Marketing, B.Pharm, D.Pharm, Google Certified Digital Marketing Professional. Qualified PhD Candidate in Operations and Management with special focus on Artificial Intelligence and Machine Learning adoption, analysis and use in Healthcare, Hospital & Pharma Domain. Seasoned with diverse therapy area of Pharmaceutical Sales & Marketing ranging from generating revenue through generating prescriptions, launching new products, and making them big brands with continuous strategy execution at the Physician and Patients level. Moved from Sales to Marketing and Business Development for 3.5 years in South East Asian Market operating from Manila, Philippines. Came back to India and handled and developed Brands such as Gluconorm, Lupisulin, Supracal, Absolut Woman, Hemozink, Fabiflu (For COVID 19), and many more. In my previous assignment I used to develop and execute strategies on Sales & Marketing, Commercialization & Business Development for Institution and Corporate Hospital Business portfolio of Oncology Therapy Area for AstraZeneca Pharma India Ltd. Being a Research Scholar and Student of ‘Operations Research & Management: Artificial Intelligence’ I published several pioneer research papers and book chapters on the same in Internationally reputed journals and Books indexed in Scopus, Springer and Ei Compendex, Google Scholar etc. Currently, I am launching PGDM Pharmaceutical Management Program in IIHMR Bangalore and spearheading the course curriculum and structure of the same. I am interested in Collaboration for Healthcare Innovation, Pharma AI Innovation, Future trend in Marketing and Management with incubation on Healthcare, Healthcare IT startups, AI-ML Modelling and Healthcare Algorithm based training module development. I am also an affiliated member of the Institute of Management Consultant of India, looking forward to Healthcare, Healthcare IT and Innovation, Pharma and Hospital Management Consulting works.",institutionString:null,institution:{name:"Lovely Professional University",country:{name:"India"}}},{id:"1063",title:"Prof.",name:"Constantin",middleName:null,surname:"Volosencu",slug:"constantin-volosencu",fullName:"Constantin Volosencu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/1063/images/system/1063.png",biography:"Prof. Dr. Constantin Voloşencu graduated as an engineer from\nPolitehnica University of Timișoara, Romania, where he also\nobtained a doctorate degree. He is currently a full professor in\nthe Department of Automation and Applied Informatics at the\nsame university. Dr. Voloşencu is the author of ten books, seven\nbook chapters, and more than 160 papers published in journals\nand conference proceedings. He has also edited twelve books and\nhas twenty-seven patents to his name. He is a manager of research grants, editor in\nchief and member of international journal editorial boards, a former plenary speaker, a member of scientific committees, and chair at international conferences. His\nresearch is in the fields of control systems, control of electric drives, fuzzy control\nsystems, neural network applications, fault detection and diagnosis, sensor network\napplications, monitoring of distributed parameter systems, and power ultrasound\napplications. He has developed automation equipment for machine tools, spooling\nmachines, high-power ultrasound processes, and more.",institutionString:"Polytechnic University of Timişoara",institution:{name:"Polytechnic University of Timişoara",country:{name:"Romania"}}},{id:"221364",title:"Dr.",name:"Eneko",middleName:null,surname:"Osaba",slug:"eneko-osaba",fullName:"Eneko Osaba",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/221364/images/system/221364.jpg",biography:"Dr. Eneko Osaba works at TECNALIA as a senior researcher. He obtained his Ph.D. in Artificial Intelligence in 2015. He has participated in more than twenty-five local and European research projects, and in the publication of more than 130 papers. He has performed several stays at universities in the United Kingdom, Italy, and Malta. Dr. Osaba has served as a program committee member in more than forty international conferences and participated in organizing activities in more than ten international conferences. He is a member of the editorial board of the International Journal of Artificial Intelligence, Data in Brief, and Journal of Advanced Transportation. He is also a guest editor for the Journal of Computational Science, Neurocomputing, Swarm, and Evolutionary Computation and IEEE ITS Magazine.",institutionString:"TECNALIA Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"275829",title:"Dr.",name:"Esther",middleName:null,surname:"Villar-Rodriguez",slug:"esther-villar-rodriguez",fullName:"Esther Villar-Rodriguez",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/275829/images/system/275829.jpg",biography:"Dr. Esther Villar obtained a Ph.D. in Information and Communication Technologies from the University of Alcalá, Spain, in 2015. She obtained a degree in Computer Science from the University of Deusto, Spain, in 2010, and an MSc in Computer Languages and Systems from the National University of Distance Education, Spain, in 2012. Her areas of interest and knowledge include natural language processing (NLP), detection of impersonation in social networks, semantic web, and machine learning. Dr. Esther Villar made several contributions at conferences and publishing in various journals in those fields. Currently, she is working within the OPTIMA (Optimization Modeling & Analytics) business of TECNALIA’s ICT Division as a data scientist in projects related to the prediction and optimization of management and industrial processes (resource planning, energy efficiency, etc).",institutionString:"TECNALIA Research & Innovation",institution:{name:"Tecnalia",country:{name:"Spain"}}},{id:"49813",title:"Dr.",name:"Javier",middleName:null,surname:"Del Ser",slug:"javier-del-ser",fullName:"Javier Del Ser",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/49813/images/system/49813.png",biography:"Prof. Dr. Javier Del Ser received his first PhD in Telecommunication Engineering (Cum Laude) from the University of Navarra, Spain, in 2006, and a second PhD in Computational Intelligence (Summa Cum Laude) from the University of Alcala, Spain, in 2013. He is currently a principal researcher in data analytics and optimisation at TECNALIA (Spain), a visiting fellow at the Basque Center for Applied Mathematics (BCAM) and a part-time lecturer at the University of the Basque Country (UPV/EHU). His research interests gravitate on the use of descriptive, prescriptive and predictive algorithms for data mining and optimization in a diverse range of application fields such as Energy, Transport, Telecommunications, Health and Industry, among others. In these fields he has published more than 240 articles, co-supervised 8 Ph.D. theses, edited 6 books, coauthored 7 patents and participated/led more than 40 research projects. He is a Senior Member of the IEEE, and a recipient of the Biscay Talent prize for his academic career.",institutionString:"Tecnalia Research & Innovation",institution:null},{id:"278948",title:"Dr.",name:"Carlos Pedro",middleName:null,surname:"Gonçalves",slug:"carlos-pedro-goncalves",fullName:"Carlos Pedro Gonçalves",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRcmyQAC/Profile_Picture_1564224512145",biography:'Carlos Pedro Gonçalves (PhD) is an Associate Professor at Lusophone University of Humanities and Technologies and a researcher on Complexity Sciences, Quantum Technologies, Artificial Intelligence, Strategic Studies, Studies in Intelligence and Security, FinTech and Financial Risk Modeling. He is also a progammer with programming experience in:\n\nA) Quantum Computing using Qiskit Python module and IBM Quantum Experience Platform, with software developed on the simulation of Quantum Artificial Neural Networks and Quantum Cybersecurity;\n\nB) Artificial Intelligence and Machine learning programming in Python;\n\nC) Artificial Intelligence, Multiagent Systems Modeling and System Dynamics Modeling in Netlogo, with models developed in the areas of Chaos Theory, Econophysics, Artificial Intelligence, Classical and Quantum Complex Systems Science, with the Econophysics models having been cited worldwide and incorporated in PhD programs by different Universities.\n\nReceived an Arctic Code Vault Contributor status by GitHub, due to having developed open source software preserved in the \\"Arctic Code Vault\\" for future generations (https://archiveprogram.github.com/arctic-vault/), with the Strategy Analyzer A.I. module for decision making support (based on his PhD thesis, used in his Classes on Decision Making and in Strategic Intelligence Consulting Activities) and QNeural Python Quantum Neural Network simulator also preserved in the \\"Arctic Code Vault\\", for access to these software modules see: https://github.com/cpgoncalves. He is also a peer reviewer with outsanding review status from Elsevier journals, including Physica A, Neurocomputing and Engineering Applications of Artificial Intelligence. Science CV available at: https://www.cienciavitae.pt//pt/8E1C-A8B3-78C5 and ORCID: https://orcid.org/0000-0002-0298-3974',institutionString:"University of Lisbon",institution:{name:"Universidade Lusófona",country:{name:"Portugal"}}},{id:"241400",title:"Prof.",name:"Mohammed",middleName:null,surname:"Bsiss",slug:"mohammed-bsiss",fullName:"Mohammed Bsiss",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/241400/images/8062_n.jpg",biography:null,institutionString:null,institution:null},{id:"276128",title:"Dr.",name:"Hira",middleName:null,surname:"Fatima",slug:"hira-fatima",fullName:"Hira Fatima",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/276128/images/14420_n.jpg",biography:"Dr. Hira Fatima\nAssistant Professor\nDepartment of Mathematics\nInstitute of Applied Science\nMangalayatan University, Aligarh\nMobile: no : 8532041179\nhirafatima2014@gmal.com\n\nDr. Hira Fatima has received his Ph.D. degree in pure Mathematics from Aligarh Muslim University, Aligarh India. Currently working as an Assistant Professor in the Department of Mathematics, Institute of Applied Science, Mangalayatan University, Aligarh. She taught so many courses of Mathematics of UG and PG level. Her research Area of Expertise is Functional Analysis & Sequence Spaces. She has been working on Ideal Convergence of double sequence. She has published 17 research papers in National and International Journals including Cogent Mathematics, Filomat, Journal of Intelligent and Fuzzy Systems, Advances in Difference Equations, Journal of Mathematical Analysis, Journal of Mathematical & Computer Science etc. She has also reviewed few research papers for the and international journals. She is a member of Indian Mathematical Society.",institutionString:null,institution:null},{id:"414880",title:"Dr.",name:"Maryam",middleName:null,surname:"Vatankhah",slug:"maryam-vatankhah",fullName:"Maryam Vatankhah",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Borough of Manhattan Community College",country:{name:"United States of America"}}},{id:"414879",title:"Prof.",name:"Mohammad-Reza",middleName:null,surname:"Akbarzadeh-Totonchi",slug:"mohammad-reza-akbarzadeh-totonchi",fullName:"Mohammad-Reza Akbarzadeh-Totonchi",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Ferdowsi University of Mashhad",country:{name:"Iran"}}},{id:"414878",title:"Prof.",name:"Reza",middleName:null,surname:"Fazel-Rezai",slug:"reza-fazel-rezai",fullName:"Reza Fazel-Rezai",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"American Public University System",country:{name:"United States of America"}}},{id:"302698",title:"Dr.",name:"Yao",middleName:null,surname:"Shan",slug:"yao-shan",fullName:"Yao Shan",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Dalian University of Technology",country:{name:"China"}}},{id:"125911",title:"Prof.",name:"Jia-Ching",middleName:null,surname:"Wang",slug:"jia-ching-wang",fullName:"Jia-Ching Wang",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"National Central University",country:{name:"Taiwan"}}},{id:"357085",title:"Mr.",name:"P. Mohan",middleName:null,surname:"Anand",slug:"p.-mohan-anand",fullName:"P. Mohan Anand",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Indian Institute of Technology Kanpur",country:{name:"India"}}},{id:"356696",title:"Ph.D. Student",name:"P.V.",middleName:null,surname:"Sai Charan",slug:"p.v.-sai-charan",fullName:"P.V. Sai Charan",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Indian Institute of Technology Kanpur",country:{name:"India"}}},{id:"357086",title:"Prof.",name:"Sandeep K.",middleName:null,surname:"Shukla",slug:"sandeep-k.-shukla",fullName:"Sandeep K. 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He obtained his Master’s degree in the Department of Information and Communications from Gwangju Institute of Science and Technology (GIST) in 2003. In 2010, he received his Ph.D. degree in the School of Information and Mechatronics from GIST. In the meantime, he was an executed team leader at Culture Technology Institute, GIST, 2010-2012. In 2011, he worked at Lancaster University, the UK as a visiting scholar. In September 2012, he joined Daegu University, where he is currently an associate professor in the School of ICT Conver, Daegu University. Also, he served as the Board of Directors of KSIIS since 2019, and HCI Korea since 2016. From 2017~2019, he worked as a center director of the Mixed Reality Convergence Research Center at Daegu University. From 2015-2017, He worked as a director in the Enterprise Supporting Office of LINC Project Group, Daegu University. 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