Search results for: Encoding

Authors: Mahbuba Rahman, Sabri Boughorbel, Scott Presnell, Charlie Quinn, Darawan Rinchai, Damien Chaussabel, Nico Marr

Abstract:

Collections of large-scale datasets made available in public repositories can be used to identify and fill gaps in biomedical knowledge. But first, these data need to be made readily accessible to researchers for analysis and interpretation. Here a collection of transcriptome datasets was made available to investigate the functional programming of human hematopoietic cells in early life. Thirty two datasets were retrieved from the NCBI Gene Expression Omnibus (GEO) and loaded in a custom, interactive web application called the Gene Expression browser (GXB), designed for visualization and query of integrated large-scale data. Multiple sample groupings and gene rank lists were created based on the study design and variables in each dataset. Web links to customized graphical views can be generated by users and subsequently be used to graphically present data in manuscripts for publication. The GXB tool also enables browsing of a single gene across datasets, which can provide information on the role of a given molecule across biological systems. The dataset collection is available online. As a proof-of-principle, one of the datasets (GSE25087) was re-analyzed to identify genes that are differentially expressed by regulatory T cells in early life. Re-analysis of this dataset and a cross-study comparison using multiple other datasets in the above mentioned collection revealed that PMCH, a gene encoding a precursor of melanin-concentrating hormone (MCH), a cyclic neuropeptide, is highly expressed in a variety of other hematopoietic cell types, including neonatal erythroid cells as well as plasmacytoid dendritic cells upon viral infection. Our findings suggest an as yet unrecognized role of MCH in immune regulation, thereby highlighting the unique potential of the curated dataset collection and systems biology approach to generate new hypotheses which can be tested in future mechanistic studies.

Keywords: early-life, GEO datasets, PMCH, interactive query, systems biology

Procedia PDF Downloads 296

Authors: Aleya Ferdausi, Meriel Jones, Anthony Halls

Abstract:

The Amaryllidaceae genus Narcissus contains secondary metabolites, which are important sources of bioactive compounds such as pharmaceuticals indicating that their biological activity extends from the native plant to humans. Transcriptome analysis (RNA-seq) is an effective platform for the identification and functional characterization of candidate genes as well as to identify genes encoding uncharacterized enzymes. The biotechnological production of secondary metabolites in plant cell or organ cultures has become a tempting alternative to the extraction of whole plant material. The biochemical pathways for the production of secondary metabolites require primary metabolites to undergo a series of modifications catalyzed by enzymes such as cytochrome P450s, methyltransferases, glycosyltransferases, and acyltransferases. Differential gene expression analysis of Narcissus was obtained from two conditions, i.e. field and in vitro callus. Callus was obtained from modified MS (Murashige and Skoog) media supplemented with growth regulators and twin-scale explants from Narcissus cv. Carlton bulb. A total of 2153 differentially expressed transcripts were detected in Narcissus bulb and in vitro callus, and 78.95% of those were annotated. It showed the expression of genes involved in the biosynthesis of alkaloids were present in both conditions i.e. cytochrome P450s, O-methyltransferase (OMTs), NADP/NADPH dehydrogenases or reductases, SAM-synthetases or decarboxylases, 3-ketoacyl-CoA, acyl-CoA, cinnamoyl-CoA, cinnamate 4-hydroxylase, alcohol dehydrogenase, caffeic acid, N-methyltransferase, and NADPH-cytochrome P450s. However, cytochrome P450s and OMTs involved in the later stage of Amaryllidaceae alkaloids biosynthesis were mainly up-regulated in field samples. Whereas, the enzymes involved in initial biosynthetic pathways i.e. fructose biphosphate adolase, aminotransferases, dehydrogenases, hydroxyl methyl glutarate and glutamate synthase leading to the biosynthesis of precursors; tyrosine, phenylalanine and tryptophan for secondary metabolites were up-regulated in callus. The knowledge of probable genes involved in secondary metabolism and their regulation in different tissues will provide insight into the Narcissus plant biology related to alkaloid production.

Keywords: narcissus, callus, transcriptomics, secondary metabolites

Procedia PDF Downloads 143

Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender

Abstract:

The ability of yeast cell wall-derived mannoproteins (glycoproteins) to positively contribute to oenological properties has been a key factor that stimulates research initiatives into these industrially important glycoproteins. In addition, and from a fundamental research perspective, yeast cell wall glycoproteins are involved in a wide range of biological interactions. To date, and to the best of our knowledge, our understanding of the fine molecular structure of these mannoproteins is fairly limited. Generally, the amino acid sequences of their protein moieties have been established from structural and functional analysis of the genomic sequence of these yeasts whilst far less information is available on the glycosyl moieties of these mannoproteins. A novel strategy was devised in this study that entails the genetic engineering of yeast strains that over-express and release cell wall-associated glycoproteins into the liquid growth medium. To this end, the Flo1p mannoprotein was overexpressed in Saccharomyces cerevisiae laboratory strains bearing a specific deletion in KNR4 and GPI7 genes involved in cell wall biosynthesis that have been previously shown to extracellularly hyper-secrete cell wall-associated glycoproteins. A polymerase chain reaction (PCR) -based cloning strategy was employed to generate transgenic yeast strains in which the native cell wall FLO1 glycoprotein-encoding gene is brought under transcriptional control of the constitutive PGK1 promoter. The modified Helm’s flocculation assay was employed to assess flocculation intensities of a Flo1p over-expressing wild type and deletion mutant as an indirect measure of their abilities to release the desired mannoprotein. The flocculation intensities of the transformed strains were assessed and all the strains showed similar intensities (>98% flocculation). To assess if mannoproteins were released into the growth medium, the supernatant of each strain was subjected to the BCA protein assay and the transformed Δknr4 strain showed a considerable increase in protein levels. This study has the potential to produce mannoproteins in sufficient quantities that may be employed in future investigations to understand their molecular structures and mechanisms of interaction to the benefit of both fundamental and industrial applications.

Keywords: glycoproteins, genetic engineering, flocculation, over-expression

Procedia PDF Downloads 415

Authors: I. Wiedemann, A. R. Sharifi, A. Mählmeyer, C. Knorr

Abstract:

A requirement for sperm motility is a morphologically intact flagellum with a central axoneme. The flagellar beating is caused by the varying activation and inactivation of dynein molecules which are located in the axoneme. DNAL4 (dynein, axonemal, light chain 4) is regarded as a possible functional candidate gene encoding a small subunit of the dyneins. In the present study, 5814bp of the porcine DNAL4 (GenBank Acc. No. AM284696.1, 6097 bp, 4 exons) were comparatively sequenced using three boars with a high motility (>68%) and three with a low motility (<60%). Primers were self-designed except for those covering exons 1, 2 and 3. Prior to sequencing, the PCR products were purified. Sequencing was performed with an ABI PRISM 3100 Genetic Analyzer using the BigDyeTM Terminator v3.1 Cycle Sequencing Reaction Kit. Finally, 23 SNPs were described and genotyped for 82 AI boars representing the breeds Piétrain, German Large White and German Landrace. The genotypes were used to assess possible associations with standard spermatological parameters (ejaculate volume, density, and sperm motility (undiluted (Motud), 24h (Mot1) and 48h (Mot2) after semen collection) that were regularly recorded on the AI station. The analysis included a total of 8,833 spermatological data sets which ranged from 2 to 295 sets per boar in five years. Only SNP g.1007A>G had a significant effect. Finally, the gene substitution effect using the following statistical model was calculated: Yijk= µ+αi+βj+αβij+b1Sijk+b2Aijk+b3T ijk + b4Vijk+b5(α*A)ijk +b6(β*A)ijk+b7(A*T)ijk+Uijk+eijk where Yijk is the semen characteristics, µ is the general mean, α is the main effect of breed, β is the main effect of season, S is the effect of SNP (g.1007A > G), A is the effect of age at semen collection, V is the effect of diluter, αβ, α*A, β*A, A*T are interactions between the fixed effects, b1-b7 are regression coefficients between y and the respective covariate, U is the random effect of repeated observation on animal and e is the random error. The results from the single marker regression analysis revealed highly significant effects (p < 0.0001) of SNP g.1007A > G on Mot1 resp. on Mot2, resulting in a marked reduction by 11.4% resp. 15.4%. Furthermore a loss of Motud by 4.6% was detected (p < 0.0178). Considering the SNP g.1007A > G as a main factor (dominant-recessive model), significant differences between genotypes AA and AG as well as AA and GG for Mot1 and Mot2 exist. For Motud there was a significant difference between AA and GG.

Keywords: association, DNAL4, porcine, sperm traits

Procedia PDF Downloads 460

Authors: Nawras Kurzom, Avi Mendelsohn

Abstract:

The effects of background music on learning and memory are inconsistent, partly due to the intrinsic complexity and variety of music and partly to individual differences in music perception and preference. A prominent musical feature that is known to elicit strong emotional responses is musical tension. Musical tension can be brought about by building anticipation of rhythm, harmony, melody, and dynamics. Delaying the resolution of dominant-to-tonic chord progressions, as well as using dissonant harmonics, can elicit feelings of tension, which can, in turn, affect memory formation of concomitant information. The aim of the presented studies was to explore how forming declarative memory is influenced by musical tension, brought about within continuous music as well as in the form of isolated chords with varying degrees of dissonance/consonance. The effects of musical tension on long-term memory of declarative information were studied in two ways: 1) by evoking tension within continuous music pieces by delaying the release of harmonic progressions from dominant to tonic chords, and 2) by using isolated single complex chords with various degrees of dissonance/roughness. Musical tension was validated through subjective reports of tension, as well as physiological measurements of skin conductance response (SCR) and pupil dilation responses to the chords. In addition, music information retrieval (MIR) was used to quantify musical properties associated with tension and its release. Each experiment included an encoding phase, wherein individuals studied stimuli (words or images) with different musical conditions. Memory for the studied stimuli was tested 24 hours later via recognition tasks. In three separate experiments, we found positive relationships between tension perception and physiological measurements of SCR and pupil dilation. As for memory performance, we found that background music, in general, led to superior memory performance as compared to silence. We detected a trade-off effect between tension perception and memory, such that individuals who perceived musical tension as such displayed reduced memory performance for images encoded during musical tension, whereas tense music benefited memory for those who were less sensitive to the perception of musical tension. Musical tension exerts complex interactions with perception, emotional responses, and cognitive performance on individuals with and without musical training. Delineating the conditions and mechanisms that underlie the interactions between musical tension and memory can benefit our understanding of musical perception at large and the diverse effects that music has on ongoing processing of declarative information.

Keywords: musical tension, declarative memory, learning and memory, musical perception

Procedia PDF Downloads 98

Authors: Belbaly Nassim

Abstract:

Virtual teams (VT) initiatives, in which teams are geographically dispersed and communicate via modern computer-driven technologies, have attracted increasing attention from researchers and professionals. The growing need to examine how to balance and optimize VT is particularly important given the exposure experienced by companies when their employees encounter globalization and decentralization pressures to monitor VT performance. Hence, organization is regularly limited due to misalignment between the behavioral capabilities of the team’s dispersed competences and knowledge capabilities and how trust issues interplay and influence these VT dimensions and the effects of such exchanges. In fact, the future success of business depends on the extent to which VTs are managing efficiently their dispersed expertise, skills and knowledge to stimulate VT creativity. Transactive memory system (TMS) may enhance VT creativity using its three dimensons: knowledge specialization, credibility and knowledge coordination. TMS can be understood as a composition of both a structural component residing of individual knowledge and a set of communication processes among individuals. The individual knowledge is shared while being retrieved, applied and the learning is coordinated. TMS is driven by the central concept that the system is built on the distinction between internal and external memory encoding. A VT learns something new and catalogs it in memory for future retrieval and use. TMS uses the role of information technology to explain VT behaviors by offering VT members the possibility to encode, store, and retrieve information. TMS considers the members of a team as a processing system in which the location of expertise both enhances knowledge coordination and builds trust among members over time. We build on TMS dimensions to hypothesize the effects of specialization, coordination, and credibility on VT creativity. In fact, VTs consist of dispersed expertise, skills and knowledge that can positively enhance coordination and collaboration. Ultimately, this team composition may lead to recognition of both who has expertise and where that expertise is located; over time, the team composition may also build trust among VT members over time developing the ability to coordinate their knowledge which can stimulate creativity. We also assess the reciprocal relationship between TMS dimensions and VT creativity. We wish to use TMS to provide researchers with a theoretically driven model that is empirically validated through survey evidence. We propose that TMS provides a new way to enhance and balance VT creativity. This study also provides researchers insight into the use of TMS to influence positively VT creativity. In addition to our research contributions, we provide several managerial insights into how TMS components can be used to increase performance within dispersed VTs.

Keywords: virtual team creativity, transactive memory systems, specialization, credibility, coordination

Procedia PDF Downloads 173

Authors: Hani M. Alothaid, Louise Robson, Richmond Muimo

Abstract:

Cystic fibrosis (CF) is a disease that affects respiratory function and in EU it affects about 1 in 2,500 live births with an average 40-year life expectancy. This disease caused by mutations within the gene encoding the CFTR (Cystic Fibrosis Transmembrane Conductance Regulator) chloride channel leading to dysregulation of epithelial fluid transport and chronic lung inflammation, suggesting functional alterations of immune cells. In airways, CFTR been found to form a functional complex with S100A10 and AnxA2 in a cAMP/PKA dependent manner. The multiprotein complex of AnxA2-S100A10 and CFTR is also regulated by calcineurin. The aim of this study was i) to investigate whether chloride ion (Cl−) channels are activated by Pseudomonas aeruginosa lipopolysaccharide (LPS from PA), ii) if this activation is regulated by cAMP/PKA/calcineurin pathway and iii) to investigate the role of LPS-activated Cl− channels in the release of pro-inflammatory cytokines by immune cells. Human peripheral blood monocytes were used in the study. Whole-cell patch records showed that LPS from PA can activate Cl− channels, including CFTR and outwardly-rectifying Cl− channel (ORCC). This activation appears to require an intact PKA/calcineurin signalling pathway. The Gout in the presence of LPS was significantly inhibited by diisothiocyanatostilbene-disulfonic acid (DIDS), an ORCC blocker (p<0.001). The Gout was further suppressed by CFTR(inh)-172, a specific inhibitor for CFTR channels (p<0.001). Monocytes pre-incubated with PKA inhibitor or calcineurin inhibitor before stimulated with LPS from PA that were resulted in DIDS and CFTR(inh)-172 insensitive currents. Activation of both ORCC and CFTR was however, observed in response to monocytes exposure to LPS. Additionally, ELISA showed that the CFTR and ORCC play a role in mediating the release of pro-inflammatory cytokines such as IL-1β upon exposure of monocytes to LPS. However, this secretion was significantly inhibited due to CFTR and ORCC inhibition. However, Cl− may play a role in IL-1β release independent of cAMP/PKA/calcineurin signalling due to the enhancement of IL-1β secretion even when cAMP/PKA/calcineurin pathway was inhibited. In conclusion, our data confirmed that LPS from PA activates Cl− channels in human peripheral blood monocytes. Our data also confirmed that Cl− channels were involved in IL-1β release in monocytes upon exposure to LPS. However, it has been found that PKA and calcineurin does not seem to influence the Cl− dependent cytokine release.

Keywords: cystic fibrosis, CFTR, Annexin A2, S100A10, PP2B, PKA, outwardly-rectifying Cl− channel, Pseudomonas aeruginosa

Procedia PDF Downloads 177

Authors: Judit Perjessy, Zsolt Zalan, Ferenc Hegyi, Eniko Horvath-Szanics, Krisztina Takacs, Andras Nagy, Adel Klupacs, Erika Koppany-Szabo, Zhirong Wang, Kaituo Wang, Muying Du, Jianquan Kan

Abstract:

The post-harvest diseases cause great economic losses in the fruit and vegetables; the prevention of these deterioration has great importance. Against the fungi, which cause most of the diseases, are extensively used the fungicides. However, there are increasing consumer concerns over the presence of pesticide residues in food. An alternative and in recent years, increasingly studied method for the prevention of the diseases is biocontrol, where antagonistic microorganisms are used for the control of fungi. The genera of Lactobacillus is well known and extensively studied, but its applicability as biocontrol agents in post-harvest preservation of fruit and vegetables is poorly investigated. However these bacteria can be found on the surface of the plants and have great antimicrobial activity. In our study we have investigated the chitinase activity, the antifungal effect and the applicability of several Lactobacillus strains to select potential biocontrol agents. We investigated the determination of the environmental parameters of a gene (encoding chitinase) expression and we also investigated the relationship between actual antifungal activity and potential chitinase activity. Mixed cultures were also developed to enhance the antifungal activity and determined the optimal mold spore and bacteria concentration ratio for the appropriate efficacy. Five Lactobacillus strains (L. acidophilus N2, L. delbrueckii subsp. bulgaricus B397, L. sp. 2231, L. sake subsp. sake 2471, L. buchneri 1145) possess chitinase-coding gene from the 43 investigated Lactobacillus strains. Proteins with similar molecular weight and separation properties like bacterial chitinases were detected from these strains, which also possess chitin-binding property. Nevertheless, they were inactive, lacks the chitinolytic activity. In point of the cumulative activity of inhibition, our results showed that certain strains were statistically significant in a positive direction compared to other strains, e.g., L. rhamnosus VT1 and L. Casey 154 have shown great general antifungal effect against 11 molds from the genera Penicillium and Botrytis and isolated from spoiled fruit and vegetables. Also, some mixed cultures (L. rhamnosus VT1 - L. Plantarum 299v) showed significant antifungal effects against the indigenous molds on the surface of apple fruit during the industrial storage experiment. Thus, they could be promising for post-harvest biopreservation.

Keywords: biocontrol, chitinase, Lactobacillus, post-harvest

Procedia PDF Downloads 154

Authors: Boutheina Ben Abdelmoumen Mardassi, Salim Chibani, Safa Boujemaa, Amaury Vaysse, Julien Guglielmini, Elhem Yacoub

Abstract:

Background and aim: Mycoplasma hominis (MH) is a pathogenic bacterium belonging to the Mollicutes class. It causes a wide range of gynecological infections and infertility among adults. Recently, we have explored for the first time the phylodistribution of Tunisian M. hominis clinical strains using an expanded MLST. We have demonstrated their distinction into two pure lineages, which each corresponding to a specific pathotype: genital infections and infertility. The aim of this project is to gain further insight into the evolutionary dynamics and the specific genetic factors that distinguish MH pathotypes Methods: Whole genome sequencing of Mycoplasma hominis clinical strains was performed using illumina Miseq. Denovo assembly was performed using a publicly available in-house pipeline. We used prokka to annotate the genomes, panaroo to generate the gene presence matrix and Jolytree to establish the phylogenetic tree. We used treeWAS to identify genetic loci associated with the pathothype of interest from the presence matrix and phylogenetic tree. Results: Our results revealed a clear categorization of the 62 MH clinical strains into two distinct genetic lineages, with each corresponding to a specific pathotype.; gynecological infections and infertility[AV1] . Genome annotation showed that GC content is ranging between 26 and 27%, which is a known characteristic of Mycoplasma genome. Housekeeping genes belonging to the core genome are highly conserved among our strains. TreeWas identified 4 virulence genes associated with the pathotype gynecological infection. encoding for asparagine--tRNA ligase, restriction endonuclease subunit S, Eco47II restriction endonuclease, and transcription regulator XRE (involved in tolerance to oxidative stress). Five genes have been identified that have a statistical association with infertility, tow lipoprotein, one hypothetical protein, a glycosyl transferase involved in capsule synthesis, and pyruvate kinase involved in biofilm formation. All strains harbored an efflux pomp that belongs to the family of multidrug resistance ABC transporter, which confers resistance to a wide range of antibiotics. Indeed many adhesion factors and lipoproteins (p120, p120', p60, p80, Vaa) have been checked and confirmed in our strains with a relatively 99 % to 96 % conserved domain and hypervariable domain that represent 1 to 4 % of the reference sequence extracted from gene bank. Conclusion: In summary, this study led to the identification of specific genetic loci associated with distinct pathotypes in M hominis.

Keywords: mycoplasma hominis, infertility, gynecological infections, virulence genes, antibiotic resistance

Procedia PDF Downloads 96

Authors: Shraddha Rane, Richard Warren, Stephen Eyre

Abstract:

L-23 is a pro-inflammatory molecule that signals T cells to release cytokines such as IL-17A and IL-22. Psoriasis is driven by a dysregulated immune response, within which IL-23 is now thought to play a key role. Genome-wide association studies (GWAS) have identified a number of genetic risk loci that support the involvement of IL-23 signalling in psoriasis; in particular a robust susceptibility locus at a gene encoding a subunit of the IL-23 receptor (IL23R) (Stuart et al., 2015; Tsoi et al., 2012). The lead psoriasis-associated SNP rs9988642 is located approximately 500 bp downstream of IL23R but is in tight linkage disequilibrium (LD) with a missense SNP rs11209026 (R381Q) within IL23R (r2 = 0.85). The minor (G) allele of rs11209026 is present in approximately 7% of the population and is protective for psoriasis and several other autoimmune diseases including IBD, ankylosing spondylitis, RA and asthma. The psoriasis-associated missense SNP R381Q causes an arginine to glutamine substitution in a region of the IL23R protein between the transmembrane domain and the putative JAK2 binding site in the cytoplasmic portion. This substitution is expected to affect the receptor’s surface localisation or signalling ability, rather than IL23R expression. Recent studies have also identified a psoriatic arthritis (PsA)-specific signal at IL23R; thought to be independent from the psoriasis association (Bowes et al., 2015; Budu-Aggrey et al., 2016). The lead PsA-associated SNP rs12044149 is intronic to IL23R and is in LD with likely causal SNPs intersecting promoter and enhancer marks in memory CD8+ T cells (Budu-Aggrey et al., 2016). It is therefore likely that the PsA-specific SNPs affect IL23R function via a different mechanism compared with the psoriasis-specific SNPs. It could be hypothesised that the risk allele for PsA located within the IL23R promoter causes an increase IL23R expression, relative to the protective allele. An increased expression of IL23R might then lead to an exaggerated immune response. The independent genetic signals identified for psoriasis and PsA in this locus indicate that different mechanisms underlie these two conditions; although likely both affecting the function of IL23R. It is very important to further characterise these mechanisms in order to better understand how the IL-23 receptor and its downstream signalling is affected in both diseases. This will help to determine how psoriasis and PsA patients might differentially respond to therapies, particularly IL-23 biologics. To investigate this further we have developed an in vitro model using CD4 T cells which express either wild type IL23R and IL12Rβ1 or mutant IL23R (R381Q) and IL12Rβ1. Model expressing different isotypes of IL23R is also underway to investigate the effects on IL23R expression. We propose to further investigate the variants for Ps and PsA and characterise key intracellular processes related to the variants.

Keywords: IL23R, psoriasis, psoriatic arthritis, SNP

Procedia PDF Downloads 168

Authors: Christoph Opperer

Abstract:

Space syntax is a theory and method for analyzing the spatial layout of buildings and urban environments to understand how they can influence patterns of human movement, social interaction, and behavior. While direct visibility is a key factor in space syntax research, important visual information such as light, color, texture, etc., are typically not considered, even though psychological studies have shown a strong correlation to the human perceptual experience within physical space – with light and color, for example, playing a crucial role in shaping the perception of spaciousness. Furthermore, these surface properties are often the visual features that are most salient and responsible for drawing attention to certain elements within the environment. This paper explores the potential of integrating these factors into general space syntax methods and visibility-based analysis of space, particularly for architectural spatial layouts. To this end, we use a combination of geometric (isovist) and topological (visibility graph) approaches together with image-based methods, allowing a comprehensive exploration of the relationship between spatial geometry, visual aesthetics, and human experience. Custom-coded ray-tracing techniques are employed to generate spherical panorama images, encoding three-dimensional spatial data in the form of two-dimensional images. These images are then processed through computer vision algorithms to generate saliency-maps, which serve as a visual representation of areas most likely to attract human attention based on their visual properties. The maps are subsequently used to weight the vertices of isovists and the visibility graph, placing greater emphasis on areas with high saliency. Compared to traditional methods, our weighted visibility analysis introduces an additional layer of information density by assigning different weights or importance levels to various aspects within the field of view. This extends general space syntax measures to provide a more nuanced understanding of visibility patterns that better reflect the dynamics of human attention and perception. Furthermore, by drawing parallels to traditional isovist and VGA analysis, our weighted approach emphasizes a crucial distinction, which has been pointed out by Ervin and Steinitz: the difference between what is possible to see and what is likely to be seen. Therefore, this paper emphasizes the importance of including surface properties in visibility-based analysis to gain deeper insights into how people interact with their surroundings and to establish a stronger connection with human attention and perception.

Keywords: space syntax, visibility analysis, isovist, visibility graph, visual features, human perception, saliency detection, raytracing, spherical images

Procedia PDF Downloads 74

Authors: Szymon Rutkowski, Tomasz Korbak

Abstract:

We present a reinforcement-learning-based method of training neural machine translation models without parallel corpora. The standard encoder-decoder approach to machine translation suffers from two problems we aim to address. First, it needs parallel corpora, which are scarce, especially for low-resource languages. Second, it lacks psychological plausibility of learning procedure: learning a foreign language is about learning to communicate useful information, not merely learning to transduce from one language’s 'encoding' to another. We instead pose the problem of learning to translate as learning a policy in a communication game between two agents: the translator and the classifier. The classifier is trained beforehand on a natural language inference task (determining the entailment relation between a premise and a hypothesis) in the target language. The translator produces a sequence of actions that correspond to generating translations of both the hypothesis and premise, which are then passed to the classifier. The translator is rewarded for classifier’s performance on determining entailment between sentences translated by the translator to disciple’s native language. Translator’s performance thus reflects its ability to communicate useful information to the classifier. In effect, we train a machine translation model without the need for parallel corpora altogether. While similar reinforcement learning formulations for zero-shot translation were proposed before, there is a number of improvements we introduce. While prior research aimed at grounding the translation task in the physical world by evaluating agents on an image captioning task, we found that using a linguistic task is more sample-efficient. Natural language inference (also known as recognizing textual entailment) captures semantic properties of sentence pairs that are poorly correlated with semantic similarity, thus enforcing basic understanding of the role played by compositionality. It has been shown that models trained recognizing textual entailment produce high-quality general-purpose sentence embeddings transferrable to other tasks. We use stanford natural language inference (SNLI) dataset as well as its analogous datasets for French (XNLI) and Polish (CDSCorpus). Textual entailment corpora can be obtained relatively easily for any language, which makes our approach more extensible to low-resource languages than traditional approaches based on parallel corpora. We evaluated a number of reinforcement learning algorithms (including policy gradients and actor-critic) to solve the problem of translator’s policy optimization and found that our attempts yield some promising improvements over previous approaches to reinforcement-learning based zero-shot machine translation.

Keywords: agent-based language learning, low-resource translation, natural language inference, neural machine translation, reinforcement learning

Procedia PDF Downloads 128

Authors: Dino Carpentras, Paul J. Maher, Caoimhe O'Reilly, Michael Quayle

Abstract:

Opinion dynamics is a sub-field of agent-based modeling that focuses on people’s opinions and their evolutions over time. Despite the rapid increase in the number of publications in this field, it is still not clear how to apply these models to real-world scenarios. Indeed, there is no agreement on how people update their opinion while interacting. Furthermore, it is not clear if different topics will show the same dynamics (e.g., more polarized topics may behave differently). These problems are mostly due to the lack of experimental validation of the models. Some previous studies started bridging this gap in the literature by directly measuring people’s opinions before and after the interaction. However, these experiments force people to express their opinion as a number instead of using natural language (and then, eventually, encoding it as numbers). This is not the way people normally interact, and it may strongly alter the measured dynamics. Another limitation of these studies is that they usually average all the topics together, without checking if different topics may show different dynamics. In our work, we collected data from 200 participants on 5 unpolarized topics. Participants expressed their opinions in natural language (“agree” or “disagree”). We also measured the certainty of their answer, expressed as a number between 1 and 10. However, this value was not shown to other participants to keep the interaction based on natural language. We then showed the opinion (and not the certainty) of another participant and, after a distraction task, we repeated the measurement. To make the data compatible with opinion dynamics models, we multiplied opinion and certainty to obtain a new parameter (here called “continuous opinion”) ranging from -10 to +10 (using agree=1 and disagree=-1). We firstly checked the 5 topics individually, finding that all of them behaved in a similar way despite having different initial opinions distributions. This suggested that the same model could be applied for different unpolarized topics. We also observed that people tend to maintain similar levels of certainty, even when they changed their opinion. This is a strong violation of what is suggested from common models, where people starting at, for example, +8, will first move towards 0 instead of directly jumping to -8. We also observed social influence, meaning that people exposed with “agree” were more likely to move to higher levels of continuous opinion, while people exposed with “disagree” were more likely to move to lower levels. However, we also observed that the effect of influence was smaller than the effect of random fluctuations. Also, this configuration is different from standard models, where noise, when present, is usually much smaller than the effect of social influence. Starting from this, we built an opinion dynamics model that explains more than 80% of data variance. This model was also able to show the natural conversion of polarization from unpolarized states. This experimental approach offers a new way to build models grounded on experimental data. Furthermore, the model offers new insight into the fundamental terms of opinion dynamics models.

Keywords: experimental validation, micro-dynamics rule, opinion dynamics, update rule

Procedia PDF Downloads 109

Authors: Wenjun Hou, Marek Perkowski

Abstract:

The Traveling Salesman problem is famous in computing and graph theory. In short, it asks for the Hamiltonian cycle of the least total weight in a given graph with N nodes. All variations on this problem, such as those with K-bounded-degree nodes, are classified as NP-complete in classical computing. Although several papers propose theoretical high-level designs of quantum algorithms for the Traveling Salesman Problem, no quantum circuit implementation of these algorithms has been created up to our best knowledge. In contrast to previous papers, the goal of this paper is not to optimize some abstract complexity measures based on the number of oracle iterations, but to be able to evaluate the real circuit and time costs of the quantum computer. Using the emerging quantum programming language Q# developed by Microsoft, which runs quantum circuits in a quantum computer simulation, an implementation of the bounded-degree problem and its respective quantum circuit were created. To apply Grover’s algorithm to this problem, a quantum oracle was designed, evaluating the cost of a particular set of edges in the graph as well as its validity as a Hamiltonian cycle. Repeating the Grover algorithm with an oracle that finds successively lower cost each time allows to transform the decision problem to an optimization problem, finding the minimum cost of Hamiltonian cycles. N log₂ K qubits are put into an equiprobablistic superposition by applying the Hadamard gate on each qubit. Within these N log₂ K qubits, the method uses an encoding in which every node is mapped to a set of its encoded edges. The oracle consists of several blocks of circuits: a custom-written edge weight adder, node index calculator, uniqueness checker, and comparator, which were all created using only quantum Toffoli gates, including its special forms, which are Feynman and Pauli X. The oracle begins by using the edge encodings specified by the qubits to calculate each node that this path visits and adding up the edge weights along the way. Next, the oracle uses the calculated nodes from the previous step and check that all the nodes are unique. Finally, the oracle checks that the calculated cost is less than the previously-calculated cost. By performing the oracle an optimal number of times, a correct answer can be generated with very high probability. The oracle of the Grover Algorithm is modified using the recalculated minimum cost value, and this procedure is repeated until the cost cannot be further reduced. This algorithm and circuit design have been verified, using several datasets, to generate correct outputs.

Keywords: quantum computing, quantum circuit optimization, quantum algorithms, hybrid quantum algorithms, quantum programming, Grover’s algorithm, traveling salesman problem, bounded-degree TSP, minimal cost, Q# language

Procedia PDF Downloads 190

Authors: Ivonete Teresinha Schulter Buss Heidemann, Michelle Kuntz Durand, Aline Megumi Arakawa-Belaunde, Sandra Mara Corrêa, Leandro Martins Costa Do Araujo, Kamila Soares Maciel

Abstract:

Primary Health Care is defined as the level of a system of services that enables the achievement of answers to health needs. This level of care produces services and actions of attention to the person in the life cycle and in their health conditions or diseases. Primary Health Care refers to a conception of care model and organization of the health system that in Brazil seeks to reorganize the principles of the Unified Health System. This system is based on the principle of health as a citizen's right and duty of the State. Primary health care has family health as a priority strategy for its organization according to the precepts of the Unified Health System, structured in the logic of new sectoral practices, associating clinical work and health promotion. Thus, this study seeks to know the possibilities and limits of the care developed by professionals working in Primary Health Care. It was conducted by a qualitative approach of the participant action type, based on Paulo Freire's Research Itinerary, which corresponds to three moments: Thematic Investigation; Encoding and Decoding; and, Critical Unveiling. The themes were investigated in a health unit with the development of a culture circle with 20 professionals, from a municipality in southern Brazil, in the first half of 2021. The participants revealed as possibilities the involvement, bonding and strengthening of the interpersonal relationships of the professionals who work in the context of primary care. Promoting welcoming in primary care has favoured care and teamwork, as well as improved access. They also highlighted that care planning, the use of technologies in the process of communication and the orientation of the population enhances the levels of problem-solving capacity and the organization of services. As limits, the lack of professional recognition and the scarce material and human resources were revealed, conditions that generate tensions for health care. The reduction in the number of professionals and the low salary are pointed out as elements that boost the motivation of the health team for the development of the work. The participants revealed that due to COVID-19, the flow of care had as a priority the pandemic situation, which affected health care in primary care, and prevention and health promotion actions were canceled. The study demonstrated that empowerment and professional involvement are fundamental to promoting comprehensive and problem-solving care. However, limits of the teams are observed when exercising their activities, these are related to the lack of human and material resources, and the expansion of public health policies is urgent.

Keywords: health promotion, primary health care, health professionals, welcoming.

Procedia PDF Downloads 98

Authors: Jung-Hong Hong, Jing-Cen Yang, Cai-Yu Ou

Abstract:

The analysis of geographic inequality heavily relies on the use of location-enabled statistical data and quantitative measures to present the spatial patterns of the selected phenomena and analyze their differences. To protect the privacy of individual instance and link to administrative units, point-based datasets are spatially aggregated to area-based statistical datasets, where only the overall status for the selected levels of spatial units is used for decision making. The partition of the spatial units thus has dominant influence on the outcomes of the analyzed results, well known as the Modifiable Areal Unit Problem (MAUP). A new spatial reference framework, the Taiwan Geographical Statistical Classification (TGSC), was recently introduced in Taiwan based on the spatial partition principles of homogeneous consideration of the number of population and households. Comparing to the outcomes of the traditional township units, TGSC provides additional levels of spatial units with finer granularity for presenting spatial phenomena and enables domain experts to select appropriate dissemination level for publishing statistical data. This paper compares the results of respectively using TGSC and township unit on the mortality data and examines the spatial characteristics of their outcomes. For the mortality data between the period of January 1^st, 2008 and December 31^st, 2010 of the Taitung County, the all-cause age-standardized death rate (ASDR) ranges from 571 to 1757 per 100,000 persons, whereas the 2^nd dissemination area (TGSC) shows greater variation, ranged from 0 to 2222 per 100,000. The finer granularity of spatial units of TGSC clearly provides better outcomes for identifying and evaluating the geographic inequality and can be further analyzed with the statistical measures from other perspectives (e.g., population, area, environment.). The management and analysis of the statistical data referring to the TGSC in this research is strongly supported by the use of Geographic Information System (GIS) technology. An integrated workflow that consists of the tasks of the processing of death certificates, the geocoding of street address, the quality assurance of geocoded results, the automatic calculation of statistic measures, the standardized encoding of measures and the geo-visualization of statistical outcomes is developed. This paper also introduces a set of auxiliary measures from a geographic distribution perspective to further examine the hidden spatial characteristics of mortality data and justify the analyzed results. With the common statistical area framework like TGSC, the preliminary results demonstrate promising potential for developing a web-based statistical service that can effectively access domain statistical data and present the analyzed outcomes in meaningful ways to avoid wrong decision making.

Keywords: mortality map, spatial patterns, statistical area, variation

Procedia PDF Downloads 258

Authors: Walter Vera-Ortega, Idoia Busnadiego, Sam J. Wilson

Abstract:

Introduction: Human Immunodeficiency Virus type 1 (HIV-1) is responsible for the acquired immunodeficiency syndrome (AIDS), a leading cause of death worldwide infecting millions of people each year. Despite intensive research in vaccine development, therapies against HIV-1 infection are not curative, and the huge genetic variability of HIV-1 challenges to drug development. Current animal models for HIV-1 research present important limitations, impairing the progress of in vivo approaches. Macaques require a CD8+ depletion to progress to AIDS, and the maintenance cost is high. Mice are a cheaper alternative but need to be 'humanized,' and breeding is not possible. The development of an HIV-1 clone able to replicate in mice is a challenging proposal. The lack of human co-factors in mice impedes the function of the HIV-1 accessory proteins, Tat and Rev, hampering HIV-1 replication. However, Tat and Rev function can be replaced by constitutive/chimeric promoters, codon-optimized proteins and the constitutive transport element (CTE), generating a novel HIV-1 clone able to replicate in mice without disrupting the amino acid sequence of the virus. By minimally manipulating the genomic 'identity' of the virus, we propose the generation of an HIV-1 clone able to replicate in mice to assist in antiviral drug development. Methods: i) Plasmid construction: The chimeric promoters and CTE copies were cloned by PCR using lentiviral vectors as templates (pCGSW and pSIV-MPCG). Tat mutants were generated from replication competent HIV-1 plasmids (NHG and NL4-3). ii) Infectivity assays: Retroviral vectors were generated by transfection of human 293T cells and murine NIH 3T3 cells. Virus titre was determined by flow cytometry measuring GFP expression. Human B-cells (AA-2) and Hela cells (TZMbl) were used for infectivity assays. iii) Protein analysis: Tat protein expression was determined by TZMbl assay and HIV-1 capsid by western blot. Results: We have determined that NIH 3T3 cells are able to generate HIV-1 particles. However, they are not infectious, and further analysis needs to be performed. Codon-optimized HIV-1 constructs are efficiently made in 293T cells in a Tat and Rev independent manner and capable of packaging a competent genome in trans. CSGW is capable of generating infectious particles in the absence of Tat and Rev in human cells when 4 copies of the CTE are placed preceding the 3’LTR. HIV-1 Tat mutant clones encoding different promoters are functional during the first cycle of replication when Tat is added in trans. Conclusion: Our findings suggest that the development of an HIV-1 Tat-Rev independent clone is challenging but achievable aim. However, further investigations need to be developed prior presenting our HIV-1 clone as a candidate model for research.

Keywords: codon-optimized, constitutive transport element, HIV-1, long terminal repeats, research model

Procedia PDF Downloads 308

Authors: Camilla Simoncelli

Abstract:

The connection between language and cognition has been one of the main interests of linguistics from several years. According to the Sapir-Whorf Linguistic Relativity Theory, the way we perceive reality depends on the language we speak which in turn has a central role in the human cognition. This paper is in line with this research work with the aim of analyzing how language structures reflect on our cognitive abilities even in the description of space, which is generally considered as a human natural and universal domain. The main objective is to identify the differences in the encoding of spatial inclusion relationships in French and Italian speakers to make evidence that a significant variation exists at various levels even in two similar systems. Starting from the constitution a corpora, the first step of the study has been to establish the relevant complex prepositions marking an inclusion relation in French and Italian: au centre de, au cœur de, au milieu de, au sein de, à l'intérieur de and the opposition entre/parmi in French; al centro di, al cuore di, nel mezzo di, in seno a, all'interno di and the fra/tra contrast in Italian. These prepositions had been classified on the base of the type of Noun following them (e.g. mass nouns, concrete nouns, abstract nouns, body-parts noun, etc.) following the Collostructional Analysis of lexemes with the purpose of analyzing the preferred construction of each preposition comparing the relations construed. Comparing the Italian and the French results it has been possible to define the degree of representativeness of each target Noun for the chosen preposition studied. Lexicostatistics and Statistical Association Measures showed the values of attraction or repulsion between lexemes and a given preposition, highlighting which words are over-represented or under-represented in a specific context compared to the expected results. For instance, a Noun as Dibattiti has a negative value for the Italian Al cuore di (-1,91), but it has a strong positive representativeness for the corresponding French Au cœur de (+677,76). The value, positive or negative, is the result of a hypergeometric distribution law which displays the current use of some relevant nouns in relations of spatial inclusion by French and Italian speakers. Differences on the kind of location conceptualization denote syntactic and semantic constraints based on spatial features as well as on linguistic peculiarity, too. The aim of this paper is to demonstrate that the domain of spatial relations is basic to human experience and is linked to universally shared perceptual mechanisms which create mental representations depending on the language use. Therefore, linguistic coding strongly correlates with the way spatial distinctions are conceptualized for non-verbal tasks even in close language systems, like Italian and French.

Keywords: cognitive semantics, cross-linguistic variations, locational terms, non-verbal spatial representations

Procedia PDF Downloads 113

Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee

Abstract:

In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.

Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization

Procedia PDF Downloads 354

Authors: M. Nerantzaki, I. Koliakou, D. Bikiaris

Abstract:

This study evaluates the hypothesis that the incorporation of fibrous hydroxyapatite nanoparticles (nHA) with high crystallinity and high aspect ratio, synthesized by hydrothermal method, into Poly(butylene succinate) (PBSu), improves the bioactivity of the aliphatic polyester and affects new bone growth inhibiting resorption and enhancing bone formation. Hydroxyapatite nanorods were synthesized using a simple hydrothermal procedure. First, the HPO42- -containing solution was added drop-wise into the Ca2+-containing solution, while the molar ratio of Ca/P was adjusted at 1.67. The HA precursor was then treated hydrothermally at 200°C for 72 h. The resulting powder was characterized using XRD, FT-IR, TEM, and EDXA. Afterwards, PBSu nanocomposites containing 2.5wt% (nHA) were prepared by in situ polymerization technique for the first time and were examined as potential scaffolds for bone engineering applications. For comparison purposes composites containing either 2.5wt% micro-Bioglass (mBG) or 2.5wt% mBG-nHA were prepared and studied, too. The composite scaffolds were characterized using SEM, FTIR, and XRD. Mechanical testing (Instron 3344) and Contact Angle measurements were also carried out. Enzymatic degradation was studied in an aqueous solution containing a mixture of R. Oryzae and P. Cepacia lipases at 37°C and pH=7.2. In vitro biomineralization test was performed by immersing all samples in simulated body fluid (SBF) for 21 days. Biocompatibility was assessed using rat Adipose Stem Cells (rASCs), genetically modified by nucleofection with DNA encoding SB100x transposase and pT2-Venus-neo transposon expression plasmids in order to attain fluorescence images. Cell proliferation and viability of cells on the scaffolds were evaluated using fluoresce microscopy and MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay. Finally, osteogenic differentiation was assessed by staining rASCs with alizarine red using cetylpyridinium chloride (CPC) method. TEM image of the fibrous HAp nanoparticles, synthesized in the present study clearly showed the fibrous morphology of the synthesized powder. The addition of nHA decreased significantly the contact angle of the samples, indicating that the materials become more hydrophilic and hence they absorb more water and subsequently degrade more rapidly. In vitro biomineralization test confirmed that all samples were bioactive as mineral deposits were detected by X-ray diffractometry after incubation in SBF. Metabolic activity of rASCs on all PBSu composites was high and increased from day 1 of culture to day 14. On day 28 metabolic activity of rASCs cultured on samples enriched with bioceramics was significantly decreased due to possible differentiation of rASCs to osteoblasts. Staining rASCs with alizarin red after 28 days in culture confirmed our initial hypothesis as the presence of calcium was detected, suggesting osteogenic differentiation of rACS on PBSu/nHAp/mBG 2.5% and PBSu/mBG 2.5% composite scaffolds.

Keywords: biomaterials, hydroxyapatite nanorods, poly(butylene succinate), scaffolds

Procedia PDF Downloads 308

Authors: Rahul Ranjan

Abstract:

Using gender as a category to cull out historical analysis, feminist scholars have produced plethora of literature on the sexual symbolics and carnal practices of modern European empires. At a symbolic level, the penetration and conquest of faraway lands was charged with sexual significance and intrigue. The white male’s domination and possession of dark and fertile lands in Africa, Asia and the Americas offered, in Anne McClintock’s words, ‘a fantastic magic lantern of the mind onto which Europe projected its forbidden sexual desires and fears’. The politics of rape were also symbolically a question significant to the politics of empire. To the colonized subject, rape was a fearsome factor, a language that spoke of violent and voracious nature of imperial exploitation. The colonized often looked at rape as an act which colonizers used as tool of oppression. The rape as act of violence got encoded into the legal structure under the helm of Lord Macaulay in the so called ‘Age of Reform’ in 1860 under IPC (Indian penal code). Initially Lord Macaulay formed Indian Law Commission in 1837 in which he drafted a bill and defined the ‘crime of rape as sexual intercourse by a man to a woman against her will and without her consent , except in cases involving girls under nine years of age where consent was immaterial’. The modern English law of rape formulated under the colonial era introduced twofold issues to the forefront. On the one hand it deployed ‘technical experts’ who wrote textbooks of medical jurisprudence that were used as credential citation to make case more ‘objective’, while on the other hand the presumptions about barbaric subjects, the colonized women’s body that was docile which is prone to adultery reflected in cases. The untrustworthiness of native witness also remained an imperative for British jurists to put extra emphasis making ‘objective’ and ‘presumptuous’. This sort of formulation put women down on the pedestrian of justice because it disadvantaged her doubly through British legality and their thinking about the rape. The Imperial morality that acted as vanguards of women’s chastity coincided language of science propagated in the post-enlightenment which not only annulled non-conformist ideas but also made itself a hegemonic language, was often used as a tool and language in encoding of law. The medico-legal understanding of rape in the colonial India has its clear imprints in the post-colonial legality. The onus on the part of rape’s victim was dictated for the longest time and still continues does by widely referred idea that ‘there should signs, marks of resistance on the body of the victim’ otherwise it is likely to be considered consensual. Having said so, this paper looks at the textual continuity that had prolonged the colonial construct of women’s body and the self.

Keywords: body, politics, textual construct, phallocentric

Procedia PDF Downloads 377

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

Abstract:

GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

Procedia PDF Downloads 320

Authors: Reem Alajmi, Hind Al Harbi, Tahany Ayaad, Zainab Al Musawi

Abstract:

Hard ticks Hyalomma spp. (family: Ixodidae) are obligate ectoparasite in their all life stages on some domestic animals mainly camels and cattle. Ticks may lead to many economic and public health problems because of their blood feeding behavior. Also, they act as vectors for many bacterial, viral and protozoan agents which may cause serious diseases such as tick-born encephalitis, Rocky-mountain spotted fever, Q-fever and Lyme disease which can affect human and/or animals. In the present study, molecular identification of ticks that attack camels in Riyadh region, Saudi Arabia based on the partial sequence of mitochondrial 16s rRNA gene was applied. Also, the present study aims to detect natural infections of collected camel ticks with Rickessia spp. and Borelia spp. using PCR/hybridization of Citrate synthase encoding gene present in bacterial cells. Hard ticks infesting camels were collected from different camels located in a farm in Riyadh region, Saudi Arabia. Results of the present study showed that the collected specimens belong to two species: Hyalomma dromedari represent 99% of the identified specimens and Hyalomma marginatum which account for 1 % of identified ticks. The molecular identification was made through blasting the obtained sequence of this study with sequences already present and identified in GeneBank. All obtained sequences of H. dromedarii specimens showed 97-100% identity with the same gene sequence of the same species (Accession # L34306.1) which was used as a reference. Meanwhile, no intraspecific variations of H. marginatum mesured because only one specimen was collected. Results also had shown that the intraspecific variability between individuals of H. dromedarii obtained in 92 % of samples ranging from 0.2- 6.6%, while the remaining 7 % of the total samples of H. dromedarii showed about 10.3 % individual differences. However, the interspecific variability between H. dromedarii and H. marginatum was approximately 18.3 %. On the other hand, by using the technique of PCR/hybridization, we could detect natural infection of camel ticks with Rickettsia spp. and Borrelia spp. Results revealed the natural presence of both bacteria in collected ticks. Rickettsial spp. infection present in 29% of collected ticks, while 35% of collected specimen were infected with Borrelia spp. The valuable results obtained from the present study are a new record for the molecular identification of camel ticks in Riyadh, Saudi Arabia and their natural infection with both Rickettsia spp. and Borrelia spp. These results may help scientists to provide a good and direct control strategy of ticks in order to protect one of the most important economic animals which are camels. Also results of this project spotlight on the disease that might be transmitted by ticks to put out a direct protective plan to prevent spreading of these dangerous agents. Further molecular studies are needed to confirm the results of the present study by using other mitochondrial and nuclear genes for tick identification.

Keywords: Camel ticks, Rickessia spp. , Borelia spp. , mitochondrial 16s rRNA gene

Procedia PDF Downloads 276

Authors: Maxwell Henderson, Tristan Cook, Justin Chan Jin Le, Mark Hodson, YoungJung Chang, John Novak, Daniel Padilha, Nishan Kulatilaka, Ansu Bagchi, Sanjoy Ray, John Kelly

Abstract:

We present a method of using adiabatic quantum optimization (AQO) to solve a mixed integer nonlinear programming (MINLP) problem instance. The MINLP problem is a general form of a set of NP-hard optimization problems that are critical to many business applications. It requires optimizing a set of discrete and continuous variables with nonlinear and potentially nonconvex constraints. Obtaining an exact, optimal solution for MINLP problem instances of non-trivial size using classical computation methods is currently intractable. Current leading algorithms leverage heuristic and divide-and-conquer methods to determine approximate solutions. Creating more accurate and efficient algorithms is an active area of research. Quantum computing (QC) has several theoretical benefits compared to classical computing, through which QC algorithms could obtain MINLP solutions that are superior to current algorithms. AQO is a particular form of QC that could offer more near-term benefits compared to other forms of QC, as hardware development is in a more mature state and devices are currently commercially available from D-Wave Systems Inc. It is also designed for optimization problems: it uses an effect called quantum tunneling to explore all lowest points of an energy landscape where classical approaches could become stuck in local minima. Our work used a novel algorithm formulated for AQO to solve a special type of MINLP problem. The research focused on determining: 1) if the problem is possible to solve using AQO, 2) if it can be solved by current hardware, 3) what the currently achievable performance is, 4) what the performance will be on projected future hardware, and 5) when AQO is likely to provide a benefit over classical computing methods. Two different methods, integer range and 1-hot encoding, were investigated for transforming the MINLP problem instance constraints into a mathematical structure that can be embedded directly onto the current D-Wave architecture. For testing and validation a D-Wave 2X device was used, as well as QxBranch’s QxLib software library, which includes a QC simulator based on simulated annealing. Our results indicate that it is mathematically possible to formulate the MINLP problem for AQO, but that currently available hardware is unable to solve problems of useful size. Classical general-purpose simulated annealing is currently able to solve larger problem sizes, but does not scale well and such methods would likely be outperformed in the future by improved AQO hardware with higher qubit connectivity and lower temperatures. If larger AQO devices are able to show improvements that trend in this direction, commercially viable solutions to the MINLP for particular applications could be implemented on hardware projected to be available in 5-10 years. Continued investigation into optimal AQO hardware architectures and novel methods for embedding MINLP problem constraints on to those architectures is needed to realize those commercial benefits.

Keywords: adiabatic quantum optimization, mixed integer nonlinear programming, quantum computing, NP-hard

Procedia PDF Downloads 525

Authors: Junali Deka

Abstract:

“Pictures speak more than a thousand words”. It is the power of image which can have multiple meanings the way it is read by the viewers. This research article is an outcome of the extensive study of the phenomenon of‘selfie culture’ and dire need of self-constructed virtual identity among youths. In the recent times, there has been a revolutionary change in the concept of photography in terms of both techniques and applications. The popularity of ‘self-portraits’ mainly depend on the temporal space and time created on social networking sites like Facebook, Instagram. With reference to Stuart’s Hall encoding and decoding process, the article studies the behavior of the users who post photographs online. The photographic messages (Roland Barthes) are interpreted differently by different viewers. The notion of ‘self’, ‘self-love and practice of looking (Marita Sturken) and ways of seeing (John Berger) got new definition and dimensional together. After Oscars Night, show host Ellen DeGeneres’s selfie created the most buzz and hype in the social media. The term was judged the word of 2013, and has earned its place in the dictionary. “In November 2013, the word "selfie" was announced as being the "word of the year" by the Oxford English Dictionary. By the end of 2012, Time magazine considered selfie one of the "top 10 buzzwords" of that year; although selfies had existed long before, it was in 2012 that the term "really hit the big time an Australian origin. The present study was carried to understand the concept of ‘selfie-bug’ and the phenomenon it has created among youth (especially students) at large in developing a pseudo-image of its own. The topic was relevant and gave a platform to discuss about the cultural, psychological and sociological implications of selfie in the age of digital technology. At the first level, content analysis of the primary and secondary sources including newspapers articles and online resources was carried out followed by a small online survey conducted with the help of questionnaire to find out the student’s view on selfie and its social and psychological effects. The newspapers reports and online resources confirmed that selfie is a new trend in the digital media and it has redefined the notion of beauty and self-love. The Facebook and Instagram are the major platforms used to express one-self and creation of virtual identity. The findings clearly reflected the active participation of female students in comparison to male students. The study of the photographs of few selected respondents revealed the difference of attitude and image building among male and female users. The study underlines some basic questions about the desire of reconstruction of identity among young generation, such as - are they becoming culturally narcissist; responsible factors for cultural, social and moral changes in the society, psychological and technological effects caused by Smartphone as well, culminating into a big question mark whether the selfie is a social signifier of identity construction.

Keywords: Culture, Narcissist, Photographs, Selfie

Procedia PDF Downloads 407

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

Abstract:

Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

Procedia PDF Downloads 341

Authors: Fernanda Bravo Cornejo, Camilo Cerda Sarabia, Belén Díaz Díaz, Diego Santibañez Oyarce, Esteban Gómez Terán, Hugo Osses Prado, Raúl Caulier-Cisterna, Jorge Vergara-Quezada, Ana Moya-Beltrán

Abstract:

Streptococcus pyogenes is a gram-positive bacteria involved in a wide range of diseases and is a major-human-specific bacterial pathogen. In Chile, this year the 'Ministerio de Salud' declared an alert due to the increase in strains throughout the year. This increase can be attributed to the multitude of factors including antimicrobial resistance (AMR) and Virulence Factors (VF). Understanding these VF and AMR is crucial for developing effective strategies and improving public health responses. Moreover, experimental identification and characterization of these pathogenic mechanisms are labor-intensive and time-consuming. Therefore, new computational methods are required to provide robust techniques for accelerating this identification. Advances in Machine Learning (ML) algorithms represent the opportunity to refine and accelerate the discovery of VF associated with Streptococcus pyogenes. In this work, we evaluate the accuracy of various machine learning models in predicting the virulence factors and antimicrobial resistance of Streptococcus pyogenes, with the objective of providing new methods for identifying the pathogenic mechanisms of this organism.Our comprehensive approach involved the download of 32,798 genbank files of S. pyogenes from NCBI dataset, coupled with the incorporation of data from Virulence Factor Database (VFDB) and Antibiotic Resistance Database (CARD) which contains sequences of AMR gene sequence and resistance profiles. These datasets provided labeled examples of both virulent and non-virulent genes, enabling a robust foundation for feature extraction and model training. We employed preprocessing, characterization and feature extraction techniques on primary nucleotide/amino acid sequences and selected the optimal more for model training. The feature set was constructed using sequence-based descriptors (e.g., k-mers and One-hot encoding), and functional annotations based on database prediction. The ML models compared are logistic regression, decision trees, support vector machines, neural networks among others. The results of this work show some differences in accuracy between the algorithms, these differences allow us to identify different aspects that represent unique opportunities for a more precise and efficient characterization and identification of VF and AMR. This comparative analysis underscores the value of integrating machine learning techniques in predicting S. pyogenes virulence and AMR, offering potential pathways for more effective diagnostic and therapeutic strategies. Future work will focus on incorporating additional omics data, such as transcriptomics, and exploring advanced deep learning models to further enhance predictive capabilities.

Keywords: antibiotic resistance, streptococcus pyogenes, virulence factors., machine learning

Procedia PDF Downloads 30

Authors: Leo Nnamdi Ozurumba-Dwight

Abstract:

Messenger ribooxynucleic acid (mRNA) molecules are compositional, protein-based. These proteins, encoding mRNA molecules (which collectively connote the transcriptome), when analyzed by RNA sequencing (RNAseq), unveils the nature of gene expression in the RNA. The obtained gene expression provides clues of cellular traits and their dynamics in presentations. These can be studied in relation to function and responses. RNAseq is a practical concept in Genomics as it enables detection and quantitative analysis of mRNA molecules. Single cell and spatial transcriptomics both present varying avenues for expositions in genomic characteristics of single cells and pooled cells in disease conditions such as cancer, auto-immune diseases, hematopoietic based diseases, among others, from investigated biological tissue samples. Single cell transcriptomics helps conduct a direct assessment of each building unit of tissues (the cell) during diagnosis and molecular gene expressional studies. A typical technique to achieve this is through the use of a single-cell RNA sequencer (scRNAseq), which helps in conducting high throughput genomic expressional studies. However, this technique generates expressional gene data for several cells which lack presentations on the cells’ positional coordinates within the tissue. As science is developmental, the use of complimentary pre-established tissue reference maps using molecular and bioinformatics techniques has innovatively sprung-forth and is now used to resolve this set back to produce both levels of data in one shot of scRNAseq analysis. This is an emerging conceptual approach in methodology for integrative and progressively dependable transcriptomics analysis. This can support in-situ fashioned analysis for better understanding of tissue functional organization, unveil new biomarkers for early-stage detection of diseases, biomarkers for therapeutic targets in drug development, and exposit nature of cell-to-cell interactions. Also, these are vital genomic signatures and characterizations of clinical applications. Over the past decades, RNAseq has generated a wide array of information that is igniting bespoke breakthroughs and innovations in Biomedicine. On the other side, spatial transcriptomics is tissue level based and utilized to study biological specimens having heterogeneous features. It exposits the gross identity of investigated mammalian tissues, which can then be used to study cell differentiation, track cell line trajectory patterns and behavior, and regulatory homeostasis in disease states. Also, it requires referenced positional analysis to make up of genomic signatures that will be sassed from the single cells in the tissue sample. Given these two presented approaches to RNA transcriptomics study in varying quantities of cell lines, with avenues for appropriate resolutions, both approaches have made the study of gene expression from mRNA molecules interesting, progressive, developmental, and helping to tackle health challenges head-on.

Keywords: transcriptomics, RNA sequencing, single cell, spatial, gene expression.

Procedia PDF Downloads 122

Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay

Abstract:

Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.

Keywords: cloning, Bacillus cereus, in silico analysis, TCE

Procedia PDF Downloads 397

Authors: I. N. Nwafia, U. C. Ozumba, M. E. Ohanu, S. O. Ebede

Abstract:

Antibiotic resistance is increasing globally and has become a major health challenge. Extended-spectrum beta-lactamase is clinically important because the ESBL gene are mostly plasmid encoded and these plasmids frequently carry genes encoding resistance to other classes of antimicrobials thereby limiting antibiotic options in the treatment of infections caused by these organisms. The specific objectives of this study were to determine the prevalence of ESBLs production in Escherichia coli, to determine the antibiotic susceptibility pattern of ESBLs producing Escherichia coli, to detect TEM, SHV and CTX-M genes and the risk factors to acquisition of ESBL producing Escherichia coli. The protocol of the study was approved by Health Research and Ethics committee of the University of Nigeria Teaching Hospital (UNTH), Enugu. It was a descriptive cross-sectional study that involved all hospitalized patients in UNTH from whose specimens Escherichia coli was isolated during the period of the study. The samples analysed were urine, wound swabs, blood and cerebrospinal fluid. These samples were cultured in 5% sheep Blood agar and MacConkey agar (Oxoid Laboratories, Cambridge UK) and incubated at 35-370C for 24 hours. Escherichia coli was identified with standard biochemical tests and confirmed using API 20E auxanogram (bioMerieux, Marcy 1'Etoile, France). The antibiotic susceptibility testing was done by disc diffusion method and interpreted according to the Clinical and Laboratory Standard Institute guideline. ESBL production was confirmed using ESBL Epsilometer test strips (Liofilchem srl, Italy). The ESBL bla genes were detected with polymerase chain reaction, after extraction of DNA with plasmid mini-prep kit (Jena Bioscience, Jena, Germany). Data analysis was with appropriate descriptive and inferential statistics. One hundred and six isolates (53.00%) out of the 200 were from urine, followed by isolates from different swabs specimens 53(26.50%) and the least number of the isolates 4(2.00) were from blood (P value = 0.096). Seventy (35.00%) out of the 200 isolates, were confirmed positive for ESBL production. Forty-two (60.00%) of the isolates were from female patients while 28(40.00%) were from male patients (P value = 0.13). Sixty-eight (97.14%) of the isolates were susceptible to imipenem while all of the isolates were resistant to ampicillin, chloramphenicol and tetracycline. From the 70 positive isolates the ESBL genes detected with polymerase chain reaction were blaCTX-M (n=26; 37.14%), blaTEM (n=7; 10.00%), blaSHV (n=2; 2.86%), blaCTX-M/TEM (n=7; 10.0%), blaCTX-M/SHV (n=14; 20.0%) and blaCTX-M/TEM/SHV (n=10; 14.29%). There was no gene detected in 4(5.71%) of the isolates. The most associated risk factors to infections caused by ESBL producing Escherichia coli was previous antibiotics use for the past 3 months followed by admission in the intensive care unit, recent surgery, and urinary catheterization. In conclusion, ESBLs was detected in 4 of every 10 Escherichia coli with the predominant gene detected being CTX-M. This knowledge will enable appropriate measures towards improvement of patient health care, antibiotic stewardship, research and infection control in the hospital.

Keywords: antimicrobial, Escherichia coli, extended spectrum beta lactamase, resistance

Procedia PDF Downloads 299