Search results for: genes expression
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2477

Search results for: genes expression

647 Disruption of Cancer Cell Proliferation by Magnetic Field

Authors: Ming Ze Kao

Abstract:

Static magnetic fields (SMF) are widely used in several medical applications, especially in diagnosis of tumors. However, biological effects of the SMFs on modulating cell physiology through the Lorentz force, which is highly frequency and magnitude dependent, remain to be elucidated. Specific patterns from SMFs of static MF, delivered by means of Halbach array magnets with a gradient increment of 6.857mT/mm from center to border, were found to have profound inhibitory effect on the growth rate of human cell line derived from Nasopharyngeal carcinoma patients. The SMFs, which were shown to be noncontact, selectively impact rapid dividing cells while quiescent cells stay intact. The phenomenon acts in two modes: the arrest of cell proliferation in the G2/M phase and destruction of cell mitosis in cell division. First mode is manifested by impacting the proper formation of mitotic spindle, whereas the second results in disintegration of the cancer cell. Both modes are demonstrated when SMF was applied for 24 hours to cancer cells, the results revealed that metaphase arrest during mitosis due to activation of DNA damage response (DDR), resulting in high expression of ATM-NBS1-CHEK signaling pathways and higher G2/M phase ratio compared with control group. Here, experimental data suggest that the SMFs cause activation of cell cycle checkpoints, which implies the MFs as a potential therapeutic modality as a sensitizer for radiotherapy or chemotherapy.

Keywords: static magnetic field, DNA damage response, Halbach array, magnetic therapy

Procedia PDF Downloads 91
646 Th2 and Th17 Subsets in the Circulation of Psoriasis Patients

Authors: Chakrit Thapphan, Suteeraporn Chaowattanapanit, Sorutsiri Chareonsudjai, Wisitsak Phoksawat, Supranee Phantanawiboon, Kiatichai Faksri, Steve W. Edwards, Kanin Salao

Abstract:

Background: Psoriasis is a chronic inflammatory disease of the skin that is mediated by crosstalk between keratinocytes and immune cells, especially CD4+ T helper (Th) cells. To date, psoriasis is established as a T helper 17 (Th17) cell-mediated inflammatory process driven by the over-expression of Th17. However, the role of other CD4+T helper cells is rather controversial. Objective: Our study, thereby, aimed to characterize and analyze T cell subsets in the circulating blood of psoriasis patients and compare them to healthy controls. Methods: Peripheral blood mononuclear cells were isolated from the participants and stained with fluorescent dye-conjugated monoclonal antibodies specific for intracellular cytokines, including interferon-gamma (IFN- γ), interleukin (IL-4), IL-17 and forkhead box P3 (FOXP3), that can be used to define T helper 1 (Th1) cells, T helper 2 (Th2), T helper 17 (Th17) and regulatory T cells (Treg) respectively. Results: We found that the numbers of Th2 (59.6% ± 17.0) and Th17 (4.0% ± 2.0) cells in the circulating blood of psoriasis patients were significantly higher than those of the healthy controls (p= 0.0007 and 0.0013 respectively). In contrast, the numbers of Th1 and Treg cells were not significantly different between psoriasis patients and healthy controls (p= 0.0593 and 0.8518, respectively). Additionally, when adjusting these numbers of Th cells to Treg, we observed a similar trend that the ratio of Th2/Treg and Th17/Treg also elevated (p = 0.0007 and 0.0047, respectively). Conclusion: Taken together, our results suggest an imbalanced T exhibit toward the Th2 and Th17 skewed-immune responses in psoriasis patients.

Keywords: psoriasis, Th cell subsets, Th2 cells, Th17 cells, Treg cells

Procedia PDF Downloads 47
645 DCDNet: Lightweight Document Corner Detection Network Based on Attention Mechanism

Authors: Kun Xu, Yuan Xu, Jia Qiao

Abstract:

The document detection plays an important role in optical character recognition and text analysis. Because the traditional detection methods have weak generalization ability, and deep neural network has complex structure and large number of parameters, which cannot be well applied in mobile devices, this paper proposes a lightweight Document Corner Detection Network (DCDNet). DCDNet is a two-stage architecture. The first stage with Encoder-Decoder structure adopts depthwise separable convolution to greatly reduce the network parameters. After introducing the Feature Attention Union (FAU) module, the second stage enhances the feature information of spatial and channel dim and adaptively adjusts the size of receptive field to enhance the feature expression ability of the model. Aiming at solving the problem of the large difference in the number of pixel distribution between corner and non-corner, Weighted Binary Cross Entropy Loss (WBCE Loss) is proposed to define corner detection problem as a classification problem to make the training process more efficient. In order to make up for the lack of Dataset of document corner detection, a Dataset containing 6620 images named Document Corner Detection Dataset (DCDD) is made. Experimental results show that the proposed method can obtain fast, stable and accurate detection results on DCDD.

Keywords: document detection, corner detection, attention mechanism, lightweight

Procedia PDF Downloads 323
644 The Morality of the Sensitive in Adorno: Suffering and Recognition in the Mimesis Model

Authors: Talita Cavaignac

Abstract:

Adorno's critique of totality, especially in a split society marked by reification, also rests on the impossibility of generalizing normative principles. Given the unfeasibility of normative universalizations, which conditions can justify the possibility of criticism and normativity in Adorno's thought? If reason itself is still entangled in alienation from the model of the domination of nature, how could be possible a critical theory? In political terms, if the notion of totality is challenged by the critique of identity, how can Adorno maintain the ideal of liberation and reconciliation between private interests and the possibility of some sort of ethics without giving up a materialist theory of society and without betting in a necessary link between redemption and history? Faced with this complex of questions, it is intended to reflect on the sense in which the notion of ‘suffering’ could throw help to the epistemological problem of the foundations of criticism in Adorno's work. The idea is that, in contrast to a universalizable model of justice, Adorno mobilizes in the notion of ‘suffering’ a gateway to the critical reflection of society. He would thus develop an approach to moral problems through the sensual-bodily perspective, fear, pain, and somatic factors. Nevertheless, due to the attention to the damaged experience and to the constitution of subjectivity -a sense in which the concept of mimesis continues to stand out- we understand suffering as an expression of an objective reification. Following the statement of other authors, the intention is to think how the resources linked to the idea of ‘suffering’ in Adorno's writings are engaged in the reflection of the problem of morality and of the contradictions between universal and particular (articulated in Hegel's tradition).

Keywords: ethics, morality, sensitive, Theodor Adorno

Procedia PDF Downloads 109
643 Microbial Dark Matter Analysis Using 16S rRNA Gene Metagenomics Sequences

Authors: Hana Barak, Alex Sivan, Ariel Kushmaro

Abstract:

Microorganisms are the most diverse and abundant life forms on Earth and account for a large portion of the Earth’s biomass and biodiversity. To date though, our knowledge regarding microbial life is lacking, as it is based mainly on information from cultivated organisms. Indeed, microbiologists have borrowed from astrophysics and termed the ‘uncultured microbial majority’ as ‘microbial dark matter’. The realization of how diverse and unexplored microorganisms are, actually stems from recent advances in molecular biology, and in particular from novel methods for sequencing microbial small subunit ribosomal RNA genes directly from environmental samples termed next-generation sequencing (NGS). This has led us to use NGS that generates several gigabases of sequencing data in a single experimental run, to identify and classify environmental samples of microorganisms. In metagenomics sequencing analysis (both 16S and shotgun), sequences are compared to reference databases that contain only small part of the existing microorganisms and therefore their taxonomy assignment may reveal groups of unknown microorganisms or origins. These unknowns, or the ‘microbial sequences dark matter’, are usually ignored in spite of their great importance. The goal of this work was to develop an improved bioinformatics method that enables more complete analyses of the microbial communities in numerous environments. Therefore, NGS was used to identify previously unknown microorganisms from three different environments (industrials wastewater, Negev Desert’s rocks and water wells at the Arava valley). 16S rRNA gene metagenome analysis of the microorganisms from those three environments produce about ~4 million reads for 75 samples. Between 0.1-12% of the sequences in each sample were tagged as ‘Unassigned’. Employing relatively simple methodology for resequencing of original gDNA samples through Sanger or MiSeq Illumina with specific primers, this study demonstrates that the mysterious ‘Unassigned’ group apparently contains sequences of candidate phyla. Those unknown sequences can be located on a phylogenetic tree and thus provide a better understanding of the ‘sequences dark matter’ and its role in the research of microbial communities and diversity. Studying this ‘dark matter’ will extend the existing databases and could reveal the hidden potential of the ‘microbial dark matter’.

Keywords: bacteria, bioinformatics, dark matter, Next Generation Sequencing, unknown

Procedia PDF Downloads 216
642 Social Semiotics in the Selected Films of Chito S. Roño

Authors: Hannah Jennica P. Ello, Regina Via G. Garcia

Abstract:

Films are famous expressions of art in the country. As an expression of art, it serves as a medium in which a culture is reflected. This paper studied how films reflected the Filipino culture. In this study, social semiotics was used to analyze the semiotic resources identified in the film. The films studied were 'Feng Shui', 'Sukob', and 'The Healing', which were three of the highest grossing horror films of Chito S. Roño. The objectives of the paper were (1) to identify the semiotic resources in the film, (2) to extract their meanings, and (3) to determine how these resources were perceived in the Filipino culture. The semiotic resources identified in each film are organized into three categories: color, practices and supernatural occurrences. Each semiotic resource is analyzed through the four dimensions of social semiotics, genre, style, modality, and discourse. For color, some of the semiotic resources identified are red, white and blue; for practices, Hagiolatry, and Mariolatry, faith healing and the belief in superstitions; and for supernatural occurrences, haunting ghosts, doppelganger attacks and returning from the dead were identified. The practices that are prominent in the films are Hagiolatry and Mariolatry, belief in feng shui and belief in faith healers and albularyos. The belief of these practices shows that Filipinos have a dual faith; belief in religion and a belief in superstitions. In short, Filipinos highly practice folk Catholicism and because of this, a mixture of different cultures can be seen, as having molded the Filipino culture to what it is today.

Keywords: culture, film, semiotics, social semiotics

Procedia PDF Downloads 297
641 Association of 105A/C IL-18 Gene Single Nucleotide Polymorphism with House Dust Mite Allergy in an Atopic Filipino Population

Authors: Eisha Vienna M. Fernandez, Cristan Q. Cabanilla, Hiyasmin Lim, John Donnie A. Ramos

Abstract:

Allergy is a multifactorial disease affecting a significant proportion of the population. It is developed through the interaction of allergens and the presence of certain polymorphisms in various susceptibility genes. In this study, the correlation of the 105A/C single nucleotide polymorphism (SNP) of the IL-18 gene and house dust mite-specific IgE among Filipino allergic and non-allergic population was investigated. Atopic status was defined by serum total IgE concentration of ≥100 IU/mL, while house dust mite allergy was defined by specific IgE value ≥ +1SD of IgE of nonatopic participants. Two hundred twenty match-paired Filipino cases and controls aged 6-60 were the subjects of this investigation. The level of total IgE and Specific IgE were measured using Enzyme-Linked Immunosorbent Assay (ELISA) while Polymerase Chain Reaction – Restriction Fragment Length Polymorphism (PCR-RFLP) analysis was used in the SNP detection. Sensitization profiles of the allergic patients revealed that 97.3% were sensitized to Blomia tropicalis, 40.0% to Dermatophagoides farinae, and 29.1% to Dermatophagoides pteronyssinus. Multiple sensitization to HDMs was also observed among the 47.27% of the atopic participants. Any of the allergy classes of the atopic triad were exhibited by the cases (allergic asthma: 48.18%; allergic rhinitis: 62.73%; atopic dermatitis: 19.09%), and two or all of these atopic states are concurrently occurring in 26.36% of the cases. A greater proportion of the atopic participants with allergic asthma and allergic rhinitis were sensitized to D. farinae, and D. pteronyssinus, while more of those with atopic dermatitis were sensitized to D. pteronyssinus than D. farinae. Results show that there is overrepresentation of the allele “A” of the 105A/C IL-18 gene SNP in both cases and control groups of the population. The genotype that predominate the population is the heterozygous “AC”, followed by the homozygous wild “AA”, and the homozygous variant “CC” being the least. The study confirmed a positive association between serum specific IgE against B. tropicalis and D. pteronyssinus and the allele “C” (Bt P=0.021, Dp P=0.027) and “AC” (Bt P=0.003, Dp P=0.026) genotype. Findings also revealed that the genotypes “AA” (OR:1.217; 95% CI: 0.701-2.113) and “CC” (OR, 3.5; 95% CI: 0.727-16.849) increase the risk of developing allergy. This indicates that the 105A/C IL-18 gene SNP is a candidate genetic marker for HDM allergy among Filipino patients.

Keywords: house dust mite allergy, interleukin-18 (IL-18), single nucleotide polymorphism,

Procedia PDF Downloads 435
640 The Effect of Particulate Matter on Cardiomyocyte Apoptosis Through Mitochondrial Fission

Authors: Tsai-chun Lai, Szu-ju Fu, Tzu-lin Lee, Yuh-Lien Chen

Abstract:

There is much evidence that exposure to fine particulate matter (PM) from air pollution increases the risk of cardiovascular morbidity and mortality. According to previous reports, PM in the air enters the respiratory tract, contacts the alveoli, and enters the blood circulation, leading to the progression of cardiovascular disease. PM pollution may also lead to cardiometabolic disturbances, increasing the risk of cardiovascular disease. The effects of PM on cardiac function and mitochondrial damage are currently unknown. We used mice and rat cardiomyocytes (H9c2) as animal and in vitro cell models, respectively, to simulate an air pollution environment using PM. These results indicate that the apoptosis-related factor PUMA, a regulator of apoptosis upregulated by p53, is increased in mice treated with PM. Apoptosis was aggravated in cardiomyocytes treated with PM, as measured by TUNEL assay and Annexin V/PI. Western blot results showed that CASPASE3 was significantly increased and BCL2 (B-cell lymphoid 2) was significantly decreased under PM treatment. Concurrent exposure to PM increases mitochondrial reactive oxygen species (ROS) production by MitoSOX Red staining. Furthermore, using Mitotracker staining, PM treatment significantly shortened mitochondrial length, indicating mitochondrial fission. The expression of mitochondrial fission-related proteins p-DRP1 (phosphodynamics-related protein 1) and FIS1 (mitochondrial fission 1 protein) was significantly increased. Based on these results, the exposure to PM worsens mitochondrial function and leads to cardiomyocyte apoptosis.

Keywords: particulate matter, cardiomyocyte, apoptosis, mitochondria

Procedia PDF Downloads 73
639 Tom Stoppard: The Amorality of the Artist

Authors: Majeed Mohammed Midhin, Clare Finburgh

Abstract:

To maintain a healthy balanced loyalty between art and politics posits a debatable issue. The artist is always on the look out for the potential tension between those two realms. Therefore, one of the most painful dilemmas the artist finds is how to function in a society without sacrificing the aesthetic values of his/her work. In other words, the life-long awareness of failure which derives from the concept of the artist as caught between unflattering social realities and the need to invent genuine art forms becomes a fertilizing soil for the artists to dig deep into its origin. Thus, within the framework of this dilemma, the question of the responsibility of the artist and the relationship of the art to politics will be illuminating. The present paper tackles the idea of the amorality of the artist in selected plays by Tom Stoppard. However, Stoppard’s awareness of his situation as a refugee has led him to keep at a distance from politics. He tried hard to avoid any intervention into the realms of political debate, especially in his earliest work. On the one hand, it is not meant that he did not interest in politics as such, but rather he preferred to question it than to create a fixed ideological position. On the other hand, Stoppard’s refusal to intervene in politics is ascribed to his feeling of gratitude to Britain where he settled. As a result, Stoppard has frequently been criticized for a lack of political engagement and also for not leaning too much for the left when he does engage. His reaction to these public criticisms finds expression in his self-conscious statements which defensively stressed the artifice of his work. He, like Oscar Wilde thinks that the responsibility of the artist is devoted to the realm of his/her art. Consequently, his consciousness for the role of the artist is truly reflected in his two plays, Artist Descending a Staircase(1972) and Travesties(1974).

Keywords: amorality, dilemma, aesthetic, responsibility of the artist, political theatre

Procedia PDF Downloads 362
638 Microbial Bioagent Triggered Biochemical Response in Tea (Camellia sinensis) Inducing Resistance against Grey Blight Disease and Yield Enhancement

Authors: Popy Bora, L. C. Bora, A. Bhattacharya, Sehnaz S. Ahmed

Abstract:

Microbial bioagents, viz., Pseudomonas fluorescens, Bacillus subtilis, and Trichoderma viride were assessed for their ability to suppress grey blight caused by Pestalotiopsis theae, a major disease of tea crop in Assam. The expression of defense-related phytochemicals due to the application of these bioagents was also evaluated. The individual bioagents, as well as their combinations, were screened for their bioefficacy against P. theae in vitro using nutrient agar (NA) as basal medium. The treatment comprising a combination of the three bioagents, P. fluorescens, B. subtilis, and T. viride showed significantly the highest inhibition against the pathogen. Bioformulation of effective bioagent combinations was further evaluated under field condition, where significantly highest reduction of grey blight (90.30%), as well as the highest increase in the green leaf yield (10.52q/ha), was recorded due to application of the bioformulation containing the three bioagents. The application of the three bioformulation also recorded an enhanced level of caffeine (4.15%) and polyphenols (22.87%). A significant increase in the enzymatic activity of phenylalanine ammonia-lyase, peroxidase and polyphenol oxidase were recorded in the plants treated with the microbial bioformulation of the three bioagents. The present investigation indicates the role of microbial agents in suppressing disease, inducing plant defense response, as well as improving the quality of tea.

Keywords: enzymatic activity, grey blight, microbial bioagents, Pestalotiopsis theae, phytochemicals, plant defense, tea

Procedia PDF Downloads 113
637 Subject, Language, and Representation: Snyder's Poetics of Emptiness

Authors: Son Hyesook

Abstract:

This project explores the possibility of poetics of emptiness in the poetry of Gary Snyder, one of the most experimental American poets, interpreting his works as an expression of his Buddhist concept, emptiness. This philosophical term demonstrates the lack of intrinsic nature in all phenomena and the absence of an independent, perduring self. Snyder’s poetics of emptiness locates the extralinguistic reality, emptiness, within the contingent nexus of language itself instead of transcending or discarding it. Language, therefore, plays an important role in his poetry, a medium intentionally applied to the carrying out of this Buddhist telos. Snyder’s poetry is characterized by strangeness and disruptiveness of language as is often the case with Asian Zen discourses. The elision of a lyric ‘I’ and transitive verbs, for example, is his grammatic attempt to represent the illusory nature of the self. He replaces the solitary speaker with sparely modified, concrete but generic images to prevent any anthropocentric understanding of the world and to demonstrate human enactment into a harmonious interplay with other elements of life as a part of a vast web of interconnections, where everything is interrelated to every other thing. In many of his poems, Snyder employs grammatical and structural ellipses and paratactical construction to avoid a facile discursive relation and to help the reader illogically imagine the inexpressible, the void. Through various uses of typographical and semantical space, his poetry forces the reader to experience the ‘thought-pause’ and intuitively perceive things-as-they-are. Snyder enacts in his Poetics an alternative to postmodern perspectives on the subject, language, and representation, and revitalizes their skeptical look at any account of human agency and the possibility of language.

Keywords: subject, language, representation, poetics of emptiness

Procedia PDF Downloads 175
636 Efficient Delivery of Biomaterials into Living Organism by Using Noble Metal Nanowire Injector

Authors: Kkochorong Park, Keun Cheon Kim, Hyoban Lee, Eun Ju Lee, Bongsoo Kim

Abstract:

Introduction of biomaterials such as DNA, RNA, proteins is important for many research areas. There are many methods to introduce biomaterials into living organisms like tissue and cells. To introduce biomaterials, several indirect methods including virus‐mediated delivery, chemical reagent (i.e., lipofectamine), electrophoresis have been used. Such methods are passive delivery using an endocytosis process of cell, reducing an efficiency of delivery. Unlike the indirect delivery method, it has been reported that a direct delivery of exogenous biomolecules into nucleus have been more efficient to expression or integration of biomolecules. Nano-sized material is beneficial for detect signal from cell or deliver stimuli/materials into the cell at cellular and molecular levels, due to its similar physical scale. Especially, because 1 dimensional (1D) nanomaterials such as nanotube, nanorod and nanowire with high‐aspect ratio have nanoscale geometry and excellent mechanical, electrical, and chemical properties, they could play an important role in molecular and cellular biology. In this study, by using single crystalline 1D noble metal nanowire, we fabricated nano-sized 1D injector which can successfully interface with living cells and directly deliver biomolecules into several types of cell line (i.e., stem cell, mammalian embryo) without inducing detrimental damages on living cell. This nano-bio technology could be a promising and robust tool for introducing exogenous biomaterials into living organism.

Keywords: DNA, gene delivery, nanoinjector, nanowire

Procedia PDF Downloads 244
635 The Effect of Heart Rate and Valence of Emotions on Perceived Intensity of Emotion

Authors: Madeleine Nicole G. Bernardo, Katrina T. Feliciano, Marcelo Nonato A. Nacionales III, Diane Frances M. Peralta, Denise Nicole V. Profeta

Abstract:

This study aims to find out if heart rate variability and valence of emotion have an effect on perceived intensity of emotion. Psychology undergraduates (N = 60) from the University of the Philippines Diliman were shown 10 photographs from the Japanese Female Facial Expression (JAFFE) Database, along with a corresponding questionnaire with a Likert scale on perceived intensity of emotion. In this 3 x 2 mixed subjects factorial design, each group was either made to do a simple exercise prior to answering the questionnaire in order to increase the heart rate, listen to a heart rate of 120 bpm, or colour a drawing to keep the heart rate stable. After doing the activity, the participants then answered the questionnaire, providing a rating of the faces according to the participants’ perceived emotional intensity on the photographs. The photographs presented were either of positive or negative emotional valence. The results of the experiment showed that neither an induced fast heart rate or perceived fast heart rate had any significant effect on the participants’ perceived intensity of emotion. There was also no interaction effect of heart rate variability and valence of emotion. The insignificance of results was explained by the Philippines’ high context culture, accompanied by the prevalence of both intensely valenced positive and negative emotions in Philippine society. Insignificance in the effects were also attributed to the Cannon-Bard theory, Schachter-Singer theory and various methodological limitations.

Keywords: heart rate variability, perceived intensity of emotion, Philippines , valence of emotion

Procedia PDF Downloads 218
634 The Use of Artificial Intelligence in Diagnosis of Mastitis in Cows

Authors: Djeddi Khaled, Houssou Hind, Miloudi Abdellatif, Rabah Siham

Abstract:

In the field of veterinary medicine, there is a growing application of artificial intelligence (AI) for diagnosing bovine mastitis, a prevalent inflammatory disease in dairy cattle. AI technologies, such as automated milking systems, have streamlined the assessment of key metrics crucial for managing cow health during milking and identifying prevalent diseases, including mastitis. These automated milking systems empower farmers to implement automatic mastitis detection by analyzing indicators like milk yield, electrical conductivity, fat, protein, lactose, blood content in the milk, and milk flow rate. Furthermore, reports highlight the integration of somatic cell count (SCC), thermal infrared thermography, and diverse systems utilizing statistical models and machine learning techniques, including artificial neural networks, to enhance the overall efficiency and accuracy of mastitis detection. According to a review of 15 publications, machine learning technology can predict the risk and detect mastitis in cattle with an accuracy ranging from 87.62% to 98.10% and sensitivity and specificity ranging from 84.62% to 99.4% and 81.25% to 98.8%, respectively. Additionally, machine learning algorithms and microarray meta-analysis are utilized to identify mastitis genes in dairy cattle, providing insights into the underlying functional modules of mastitis disease. Moreover, AI applications can assist in developing predictive models that anticipate the likelihood of mastitis outbreaks based on factors such as environmental conditions, herd management practices, and animal health history. This proactive approach supports farmers in implementing preventive measures and optimizing herd health. By harnessing the power of artificial intelligence, the diagnosis of bovine mastitis can be significantly improved, enabling more effective management strategies and ultimately enhancing the health and productivity of dairy cattle. The integration of artificial intelligence presents valuable opportunities for the precise and early detection of mastitis, providing substantial benefits to the dairy industry.

Keywords: artificial insemination, automatic milking system, cattle, machine learning, mastitis

Procedia PDF Downloads 30
633 Genetic Diversity of Sugar Beet Pollinators

Authors: Ksenija Taški-Ajdukovic, Nevena Nagl, Živko Ćurčić, Dario Danojević

Abstract:

Information about genetic diversity of sugar beet parental populations is of a great importance for hybrid breeding programs. The aim of this research was to evaluate genetic diversity among and within populations and lines of diploid sugar beet pollinators, by using SSR markers. As plant material were used eight pollinators originating from three USDA-ARS breeding programs and four pollinators from Institute of Field and Vegetable Crops, Novi Sad. Depending on the presence of self-fertility gene, the pollinators were divided into three groups: autofertile (inbred lines), autosterile (open-pollinating populations), and group with partial presence of autofertility gene. A total of 40 SSR primers were screened, out of which 34 were selected for the analysis of genetic diversity. A total of 129 different alleles were obtained with mean value 3.2 alleles per SSR primer. According to the results of genetic variability assessment the number and percentage of polymorphic loci was the maximal in pollinators NS1 and tester cms2 while effective number of alleles, expected heterozygosis and Shannon’s index was highest in pollinator EL0204. Analysis of molecular variance (AMOVA) showed that 77.34% of the total genetic variation was attributed to intra-varietal variance. Correspondence analysis results were very similar to grouping by neighbor-joining algorithm. Number of groups was smaller by one, because correspondence analysis merged IFVCNS pollinators with CZ25 into one group. Pollinators FC220, FC221 and C 51 were in the next group, while self-fertile pollinators CR10 and C930-35 from USDA-Salinas were separated. On another branch were self-sterile pollinators ЕL0204 and ЕL53 from USDA-East Lansing. Sterile testers cms1 and cms2 formed separate group. The presented results confirmed that SSR analysis can be successfully used in estimation of genetic diversity within and among sugar beet populations. Since the tested pollinator differed considering the presence of self-fertility gene, their heterozygosity differed as well. It was lower in genotypes with fixed self-fertility genes. Since the most of tested populations were open-pollinated, which rarely self-pollinate, high variability within the populations was expected. Cluster analysis grouped populations according to their origin.

Keywords: auto fertility, genetic diversity, pollinator, SSR, sugar beet

Procedia PDF Downloads 431
632 Migration, Agency and Subjectivity in Helon Habila's Travellers

Authors: Bankole Wright

Abstract:

The late 20th to the early 21st century has been predominantly characterized by the movement of individuals from one country to another country or countries. The chief reasons for migration have always been premised on socio-cultural, socio-political and socio-economic factors, with influences of migration finding expression through various ways. Indeed, migration experiences have formed points of subjectivity which functions as agencies that propel migrants to strongly quest for migrating from their home space to other socio-cultural space that performs the role of escape for them. This paper interrogates the discourse of migration, agency and subjectivity in Helon Habila’s Travellers. The essay explores the interconnectedness between migration which is the physical [as deployed in this paper] movement from one location to another, agency as seen in the ability to act based on various ideological frameworks within which the action is taken, and subjectivity which identifies with the predominant factors that influence human actions; and how these connections are responsible for defining the diaspora individual. The discourse of what makes migrants desire to move from their various spaces is as critical as the experiences they face in their various host land. Hence, this paper demonstrates, through the analysis of an African diasporic novel, that the quest for migration is mostly determined by certain agencies in the diaspora home space, which characters have been subjects of and desire to escape. Traveller is a novel which chronicles the various experiences of migrants who journey from their various home space to another land as a result of different agencies that precipitated their migration. This paper engages these agencies as impediments to human survival.

Keywords: migration, agency, subjectivity, Helon Habila, diaspora, home, space

Procedia PDF Downloads 240
631 Building Deep: Mystery And Sensuality In The Underground World

Authors: Rene Davids

Abstract:

Urban undergrounds spaces such as parking garages or metro stations are perceived as interludes before reaching desired destinations, as commodities devoid of aesthetic value. Within the encoded space of the city, commercial underground spaces are the closest expression to pure to structures of consumption and commodity. Even in the house, the cellar is associated with castoffs and waste or, as scholar Mircea Eliade has pointed out at best, with a place to store abandoned household and childhood objects, which lie forgotten and on rediscovery evoke a nostalgic and uncanny sense of the past. Despite a growing body of evidence presented by an increasing number of buildings situated entirely below or semi underground that feature exemplary spatial and sensuous qualities, critics and scholars see them largely as efforts to produce efforts in producing low consumption non-renewable energy. Buildings that also free space above ground. This critical approach neglects to mention and highlight other project drivers such as the notion that the ground and sky can be considered a building’s fundamental context, that underground spaces are conducive to the exploration of pure space, namely an architecture that doesn’t have to deal with facades and or external volumes and that digging into geology can inspire the textural and spatial richness. This paper will argue that while the assessment about the reduced energy consumption of underground construction is important, it does not do justice to the qualities underground buildings can contribute to a city’s expanded urban and or landscape experiences.

Keywords: low non-renewable energy consumption, pure space, underground buildings, urban and landscape experience

Procedia PDF Downloads 156
630 Computational Identification of Signalling Pathways in Protein Interaction Networks

Authors: Angela U. Makolo, Temitayo A. Olagunju

Abstract:

The knowledge of signaling pathways is central to understanding the biological mechanisms of organisms since it has been identified that in eukaryotic organisms, the number of signaling pathways determines the number of ways the organism will react to external stimuli. Signaling pathways are studied using protein interaction networks constructed from protein-protein interaction data obtained using high throughput experimental procedures. However, these high throughput methods are known to produce very high rates of false positive and negative interactions. In order to construct a useful protein interaction network from this noisy data, computational methods are applied to validate the protein-protein interactions. In this study, a computational technique to identify signaling pathways from a protein interaction network constructed using validated protein-protein interaction data was designed. A weighted interaction graph of the Saccharomyces cerevisiae (Baker’s Yeast) organism using the proteins as the nodes and interactions between them as edges was constructed. The weights were obtained using Bayesian probabilistic network to estimate the posterior probability of interaction between two proteins given the gene expression measurement as biological evidence. Only interactions above a threshold were accepted for the network model. A pathway was formalized as a simple path in the interaction network from a starting protein and an ending protein of interest. We were able to identify some pathway segments, one of which is a segment of the pathway that signals the start of the process of meiosis in S. cerevisiae.

Keywords: Bayesian networks, protein interaction networks, Saccharomyces cerevisiae, signalling pathways

Procedia PDF Downloads 506
629 Retrospective Insight on the Changing Status of the Romanian Language Spoken in the Republic of Moldova

Authors: Gina Aurora Necula

Abstract:

From its transformation into a taboo and its hiding under the so-called “Moldovan language” or under the euphemistic expression “state language” to its regained status recognition as an official language, the Romanian language spoken in the Republic of Moldova has undergone impressive reforms in the last 60 years. Meant to erase the awareness of citizens’ ethnic identity and turn a majority language into a minority one, all the laws and regulations issued on the field succeeded into setting numerous barriers for speakers of Romanian. Either manifested as social constraints or materialized into assumed rejection of mother tongue usage, all these laws have demonstrated their usefulness and major impact on the Romanian-speaking population. This article is the result of our research carried out over 10 years with the support of students, and Moldovan citizens, from the master's degree program "Romanian language - identity and cultural awareness." We present here a retrospective insight of the reforms, laws, and regulations that contributed to the shifted status of the Romanian language from the official language, seen as the language of common use both in the public and private spheres, in the minority language that surrendered its privileged place to the Russian language, firstly in the public sphere, and then, slowly but surely, in the private sphere. Our main goal here is to identify and make speakers understand what the barriers to learning Romanian language are nowadays when the social pressure on using Russian no longer exists.

Keywords: linguistic barriers, lingua franca, private sphere, public sphere, reformation

Procedia PDF Downloads 82
628 Immunoinformatic Design and Evaluation of an Epitope-Based Tetravalent Vaccine against Human Hand, Foot, and Mouth Disease

Authors: Aliyu Maje Bello, Yaowaluck Maprang Roshorm

Abstract:

Hand, foot, and mouth disease (HFMD) is a highly contagious viral infection affecting mostly infants and children. Although the Enterovirus A71 (EV71) is usually the major causative agent of HFMD, other enteroviruses such as coxsackievirus A16, A10, and A6 are also found in some of the recent outbreaks. The commercially available vaccines have demonstrated their effectiveness against only EV71 infection but no protection against other enteroviruses. To address the limitation of the monovalent EV71 vaccine, the present study thus designed a tetravalent vaccine against the four major enteroviruses causing HFMD and primarily evaluated the designed vaccine using an immunoinformatics approach. The immunogen was designed to contain the EV71 VP1 protein and multiple reported epitopes from all four distinct enteroviruses and thus designated a tetravalent vaccine. The 3D structure of the designed tetravalent vaccine was modeled, refined, and validated. Epitope screening showed the presence of B-cell, CTL, CD4 T cell, and IFN epitopes with vast application among the Asian population. Docking analysis confirmed the stable and strong binding interactions between the immunogen and immune receptor B-cell receptor (BCR). In silico cloning and immune simulation analyses guaranteed high efficiency and sufficient expression of the vaccine candidate in humans. Overall, the promising results obtained from the in-silico studies of the proposed tetravalent vaccine make it a potential candidate worth further experimental validation.

Keywords: enteroviruses, coxsackieviruses, hand foot and mouth disease, immunoinformatics, tetravalent vaccine

Procedia PDF Downloads 35
627 Deep Vision: A Robust Dominant Colour Extraction Framework for T-Shirts Based on Semantic Segmentation

Authors: Kishore Kumar R., Kaustav Sengupta, Shalini Sood Sehgal, Poornima Santhanam

Abstract:

Fashion is a human expression that is constantly changing. One of the prime factors that consistently influences fashion is the change in colour preferences. The role of colour in our everyday lives is very significant. It subconsciously explains a lot about one’s mindset and mood. Analyzing the colours by extracting them from the outfit images is a critical study to examine the individual’s/consumer behaviour. Several research works have been carried out on extracting colours from images, but to the best of our knowledge, there were no studies that extract colours to specific apparel and identify colour patterns geographically. This paper proposes a framework for accurately extracting colours from T-shirt images and predicting dominant colours geographically. The proposed method consists of two stages: first, a U-Net deep learning model is adopted to segment the T-shirts from the images. Second, the colours are extracted only from the T-shirt segments. The proposed method employs the iMaterialist (Fashion) 2019 dataset for the semantic segmentation task. The proposed framework also includes a mechanism for gathering data and analyzing India’s general colour preferences. From this research, it was observed that black and grey are the dominant colour in different regions of India. The proposed method can be adapted to study fashion’s evolving colour preferences.

Keywords: colour analysis in t-shirts, convolutional neural network, encoder-decoder, k-means clustering, semantic segmentation, U-Net model

Procedia PDF Downloads 76
626 Association of Transforming Growth Factor-β1 Gene 1800469 C > T and 1982073 C > T Polymorphism with Type 2 Diabetic Foot Ulcer Patient in Cipto Mangunkusumo National Hospital Jakarta

Authors: Dedy Pratama, Akhmadu Muradi, Hilman Ibrahim, Patrianef Darwis, Alexander Jayadi Utama, Raden Suhartono, D. Suryandari, Luluk Yunaini, Tom Ch Adriani

Abstract:

Objective: Diabetic Foot Ulcer (DFU) is one of the complications of Type 2 Diabetes Mellitus (T2DM) that can lead to disability and death. Inadequate vascularization condition will affect healing process of DFU. Therefore, we investigated the expression of polymorphism TGF- β1 in the relation of the occurrence of DFU in T2DM. Methods: We designed a case-control study to investigate the polymorphism TGF- β1 gene 1800469 C > T and 1982073 C > T in T2DM in Cipto Mangunkusumo National Hospital (RSCM) Jakarta from June to December 2016. We used PCR techniques and compared the results in a group of T2DM patients with DFU as the case study and without DFU as the control group. Results: There were 203 patients, 102 patients with DFU and 101 patients control without DFU. 49,8% is male and 50,2% female with mean age about 56 years. Distribution of wild-type genotype TGF-B1 1800469 C > T wild type CC was found in 44,8%, the number of mutant heterozygote CT was 10,8% and mutant homozygote is 11,3%. Distribution of TGF-B1 1982073 C>T wild type CC was 32,5%, mutant heterozygote is 38,9% and mutant homozygote 25,1%. Conclusion: Distribution of alleles from TGF-B1 1800469 C > T is C 75% and T 25% and from TGF-B1 1982073 C > T is C53,8% and T 46,2%. In the other word polymorphism TGF- β1 plays a role in the occurrence and healing process of the DFU in T2DM patients.

Keywords: diabetic foot ulcers, diabetes mellitus, polymorphism, TGF-β1

Procedia PDF Downloads 255
625 Supplier Risk Management: A Multivariate Statistical Modelling and Portfolio Optimization Based Approach for Supplier Delivery Performance Development

Authors: Jiahui Yang, John Quigley, Lesley Walls

Abstract:

In this paper, the authors develop a stochastic model regarding the investment in supplier delivery performance development from a buyer’s perspective. The authors propose a multivariate model through a Multinomial-Dirichlet distribution within an Empirical Bayesian inference framework, representing both the epistemic and aleatory uncertainties in deliveries. A closed form solution is obtained and the lower and upper bound for both optimal investment level and expected profit under uncertainty are derived. The theoretical properties provide decision makers with useful insights regarding supplier delivery performance improvement problems where multiple delivery statuses are involved. The authors also extend the model from a single supplier investment into a supplier portfolio, using a Lagrangian method to obtain a theoretical expression for an optimal investment level and overall expected profit. The model enables a buyer to know how the marginal expected profit/investment level of each supplier changes with respect to the budget and which supplier should be invested in when additional budget is available. An application of this model is illustrated in a simulation study. Overall, the main contribution of this study is to provide an optimal investment decision making framework for supplier development, taking into account multiple delivery statuses as well as multiple projects.

Keywords: decision making, empirical bayesian, portfolio optimization, supplier development, supply chain management

Procedia PDF Downloads 264
624 Ellagic Acid Enhanced Apoptotic Radiosensitivity via G1 Cell Cycle Arrest and γ-H2AX Foci Formation in HeLa Cells in vitro

Authors: V. R. Ahire, A. Kumar, B. N. Pandey, K. P. Mishra, G. R. Kulkarni

Abstract:

Radiation therapy is an effective vital strategy used globally in the treatment of cervical cancer. However, radiation efficacy principally depends on the radiosensitivity of the tumor, and not all patient exhibit significant response to irradiation. A radiosensitive tumor is easier to cure than a radioresistant tumor which later advances to local recurrence and metastasis. Herbal polyphenols are gaining attention for exhibiting radiosensitization through various signaling. Current work focuses to study the radiosensitization effect of ellagic acid (EA), on HeLa cells. EA intermediated radiosensitization of HeLa cells was due to the induction γ-H2AX foci formation, G1 phase cell cycle arrest, and loss of reproductive potential, growth inhibition, drop in the mitochondrial membrane potential and protein expression studies that eventually induced apoptosis. Irradiation of HeLa in presence of EA (10 μM) to doses of 2 and 4 Gy γ-radiation produced marked tumor cytotoxicity. EA also demonstrated radio-protective effect on normal cell, NIH3T3 and aided recovery from the radiation damage. Our results advocate EA to be an effective adjuvant for improving cancer radiotherapy as it displays striking tumor cytotoxicity and reduced normal cell damage instigated by irradiation.

Keywords: apoptotic radiosensitivity, ellagic acid, mitochondrial potential, cell-cycle arrest

Procedia PDF Downloads 320
623 A Solution to Analyze the Geosynthetic Reinforced Piled Embankments Considering Pile-Soil Interaction

Authors: Feicheng Liu, Weiming Liao, Jianjing Zhang

Abstract:

A pile-supported embankment with geosynthetic-reinforced mat (PSGR embankment) has been considered as an effective solution to reduce the total and differential settlement of the embankment constructed over soft soil. In this paper, a new simplified method proposed firstly incorporates the load transfer between piles and surrounding soil and the settlement of pile, and also considers arching effect in embankment fill, membrane effect of geosynthetic reinforcement, and subsoil resistance, to evaluate the behavior of PSGR embankment. Subsoil settlement is assumed to consist of two parts:(1) the settlement of subsoil surface between piles equivalent to that of pile caps assuming the geosynthetic reinforcement without deformation yet; (2) the subsoil subsiding along with the geosynthetic deforming, and the deflected geosynthetic being considered as centenary. The force equilibrium, including loads acting on the upper surface of geosynthetic, subsoil resistance, as well as the stress-strain relationship of the geosynthetic reinforcement at the edge of pile cap, is established, thus the expression of subsoil resistance is deduced, and subsequently the tension of geosynthetic and stress concentration ratio between piles can be calculated. The proposed method is validated through observed data from three field tests and also compared with other eight analytical solutions available in the literature. In addition, a sensitive analysis is provided to demonstrate the influence of with/without considering pile-soil interaction for evaluating the performance of PSGR embankment.

Keywords: pile-supported embankment, geosynthetic, analytical solution, soil arching effect, the settlement of pile, sensitive analysis

Procedia PDF Downloads 131
622 The Inattentional Blindness Paradigm: A Breaking Wave for Attentional Biases in Test Anxiety

Authors: Kritika Kulhari, Aparna Sahu

Abstract:

Test anxiety results from concerns about failure in examinations or evaluative situations. Attentional biases are known to pronounce the symptomatic expression of test anxiety. In recent times, the inattentional blindness (IB) paradigm has shown promise as an attention bias modification treatment (ABMT) for anxiety by overcoming practice and expectancy effects which preexisting paradigms fail to counter. The IB paradigm assesses the inability of an individual to attend to a stimulus that appears suddenly while indulging in a perceptual discrimination task. The present study incorporated an IB task with three critical items (book, face, and triangle) appearing randomly in the perceptual discrimination task. Attentional biases were assessed as detection and identification of the critical item. The sample (N = 50) consisted of low test anxiety (LTA) and high test anxiety (HTA) groups based on the reactions to tests scale scores. Test threat manipulation was done with pre- and post-test assessment of test anxiety using the State Test Anxiety Inventory. A mixed factorial design with gender, test anxiety, presence or absence of test threat, and critical items was conducted to assess their effects on attentional biases. Results showed only a significant main effect for test anxiety on detection with higher accuracy of detection of the critical item for the LTA group. The study presents promising results in the realm of ABMT for test anxiety.

Keywords: attentional bias, attentional bias modification treatment, inattentional blindness, test anxiety

Procedia PDF Downloads 179
621 Research on a Digital Basketball Sports Game (DBSG) Framework Based on the Female Perspective

Authors: Ran Yue, Zhejing Li

Abstract:

Context: The context of this study is the field of Digital Basketball Sports Games (DBSG). The existing DBSGs often prioritize competitiveness and confrontation, neglecting the narrative and progressive expression, especially from a female standpoint. This study aims to address this gap by analyzing existing DBSGs and proposing a comprehensive framework tailored to meet the needs and desires of women in basketball. Research Aim: The aim of this research is to examine the narrative perspectives of women in basketball and understand their desires and expectations within the sport. It also seeks to investigate methods to seamlessly integrate women's basketball stories into gameplay, addressing their specific needs and expectations. Additionally, the study aims to develop a digital basketball sports game framework that combines narrative richness and entertainment, with a focus on the female audience. Methodology: The study utilizes affective-arousal theories as a psychological framework to explore how emotional arousal influences player engagement and responses in the digital basketball sports game. It employs in-depth case studies to examine specific instances and gain insights into the implementation and impact of narrative elements and educational features in existing DBSGs. Comparative studies are conducted to analyze different DBSGs, identifying effective strategies and shortcomings. Findings: The research findings contribute to the development of a digital basketball game framework from a female perspective. This framework enhances the completeness, diversity, and inclusivity of digital basketball sports games. By addressing the specific needs of women in basketball, including fundamental knowledge, sports skills, safety awareness, and rehabilitation training methods, the framework provides a foundational reservoir for a broader range of basketball participation. It enriches the gaming experience by enhancing enjoyment, narrative, and diversity. It also acts as a catalyst to encourage more women to engage with basketball stories, participate in the sport, persevere, and derive greater enjoyment while benefiting their physical fitness and health. Theoretical Importance: The study contributes to the existing literature by incorporating game motivation psychology theories and proposing a comprehensive framework that caters to the specific needs of women in basketball. It emphasizes the importance of considering the narrative and progressive expression in DBSGs, especially from a female perspective. The research explores affective-arousal theories and provides insights into how emotional arousal can influence player engagement and responses in digital basketball sports games. Data Collection and Analysis Procedures: The study collects data through in-depth case studies of existing DBSGs, examining specific instances to uncover insights into the implementation and impact of narrative elements and educational features. Comparative studies are conducted to contrast and analyze various DBSGs, identifying effective strategies and shortcomings. The analysis procedures involve identifying commonalities, differences, strengths, and weaknesses among the DBSGs, guiding the development of a female-centric perspective in the proposed framework. Questions Addressed: The study addresses the following questions: What are the narrative perspectives of women in basketball? How can women's basketball stories be seamlessly integrated into gameplay? What are the specific needs and expectations of women in basketball? What effective strategies and shortcomings exist in current DBSGs? How can a digital basketball game framework be developed to cater to the female audience? Conclusion: In conclusion, this study contributes to the field of DBSGs by proposing a comprehensive digital basketball game framework from a female perspective. The framework enhances the inclusivity, diversity, and enjoyment of DBSGs by addressing the specific needs and desires of women in basketball. It provides a foundation for a broader range of basketball participation, enriching the gaming experience and benefiting women's physical fitness and health. The research, using affective-arousal theories and in-depth case studies, provides valuable insights into the implementation and impact of narrative elements and educational features in existing DBSGs, guiding the development of the proposed female-centric framework.

Keywords: digital basketball game, game framework, female perspective, game narratives

Procedia PDF Downloads 25
620 Accentuation Moods of Blaming Utterances in Egyptian Arabic: A Pragmatic Study of Prosodic Focus

Authors: Reda A. H. Mahmoud

Abstract:

This paper investigates the pragmatic meaning of prosodic focus through four accentuation moods of blaming utterances in Egyptian Arabic. Prosodic focus results in various pragmatic meanings when the speaker utters the same blaming expression in different emotional moods: the angry, the mocking, the frustrated, and the informative moods. The main objective of this study is to interpret the meanings of these four accentuation moods in relation to their illocutionary forces and pre-locutionary effects, the integrated features of prosodic focus (e.g., tone movement distributions, pitch accents, lengthening of vowels, deaccentuation of certain syllables/words, and tempo), and the consonance between the former prosodic features and certain lexico-grammatical components to communicate the intentions of the speaker. The data on blaming utterances has been collected via elicitation and pre-recorded material, and the selection of blaming utterances is based on the criteria of lexical and prosodic regularity to be processed and verified by three computer programs, Praat, Speech Analyzer, and Spectrogram Freeware. A dual pragmatic approach is established to interpret expressive blaming utterance and their lexico-grammatical distributions into intonational focus structure units. The pragmatic component of this approach explains the variable psychological attitudes through the expressions of blaming and their effects whereas the analysis of prosodic focus structure is used to describe the intonational contours of blaming utterances and other prosodic features. The study concludes that every accentuation mood has its different prosodic configuration which influences the listener’s interpretation of the pragmatic meanings of blaming utterances.

Keywords: pragmatics, pragmatic interpretation, prosody, prosodic focus

Procedia PDF Downloads 120
619 Influences on Female Gender Identity and Role in Pre-School, Saudi Arabian: Analyzing Children's Perspectives through Narratives and Teachers' Pedagogies

Authors: Mona Alzahrani

Abstract:

Microworld theories can help to define the many influences on female development. In this research, theories together with narratives have been used to discover the reality of children’s gender perceptions in Saudi Arabia. Today, Saudi Arabia is considered a ‘closed and conserved’ society due to tribal, cultural and religious factors. This study focuses on how young girls in Saudi Arabia learn about what is expected of them as females. Cultural beliefs and experiences contribute to children’s notions of identity. Moreover, significant others such as more experienced peers, teachers, parents, and other members of a society can influence a child’s development of knowledge through interactions within their social world. There are dominant influences from the Saudi State. These influences have very strong devices and perceptions of what or how a female should act and be. However, children may have other viewpoints, as it also needs to be considered that the Internet and other media sources could have an influence. Consequently, difficulties could exist for these young children to feel an authentic sense of belonging. The study gathered data using a multi-method approach that elicited the perspectives of the children using ‘multiple modes of expression’ such as observations, story-telling, picture prompt cards, group interviews, drawings and annotations. For this study, prompts and a book was devised, specifically, for use in a Saudi setting. It was found that Saudi young girls in preschool were heteronomous, mainly influenced by culture and society, in their perceptions of female gender and role.

Keywords: Saudi Arabia, pre-school, female, teachers, gender, identity, role

Procedia PDF Downloads 128
618 High-Throughput Artificial Guide RNA Sequence Design for Type I, II and III CRISPR/Cas-Mediated Genome Editing

Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

Abstract:

A huge revolution has emerged in genome engineering by the discovery of CRISPR (clustered regularly interspaced palindromic repeats) and CRISPR-associated system genes (Cas) in bacteria. The function of type II Streptococcus pyogenes (Sp) CRISPR/Cas9 system has been confirmed in various species. Other S. thermophilus (St) CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been also reported for preventing phage infection. The CRISPR1-Cas system interferes by cleaving foreign dsDNA entering the cell in a length-specific and orientation-dependant manner. The S. thermophilus CRISPR3-Cas system also acts by cleaving phage dsDNA genomes at the same specific position inside the targeted protospacer as observed in the CRISPR1-Cas system. It is worth mentioning, for the effective DNA cleavage activity, RNA-guided Cas9 orthologs require their own specific PAM (protospacer adjacent motif) sequences. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of target site for the three orthogonals of Cas9 protein, a well-organized procedure will be required for high-throughput and accurate mining of possible target sites in a large genomic dataset. Consequently, we created a reliable procedure to explore potential gRNA sequences for type I (Streptococcus thermophiles), II (Streptococcus pyogenes), and III (Streptococcus thermophiles) CRISPR/Cas systems. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows: i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. The output of such procedure highlights the power of comparative genome mining for different CRISPR/Cas systems. This could yield a repertoire of Cas9 variants with expanded capabilities of gRNA design, and will pave the way for further advance genome and epigenome engineering.

Keywords: CRISPR/Cas systems, gRNA mining, Streptococcus pyogenes, Streptococcus thermophiles

Procedia PDF Downloads 219