Search results for: DNA encoding
69 Vibro-Tactile Equalizer for Musical Energy-Valence Categorization
Authors: Dhanya Nair, Nicholas Mirchandani
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Musical haptic systems can enhance a listener’s musical experience while providing an alternative platform for the hearing impaired to experience music. Current music tactile technologies focus on representing tactile metronomes to synchronize performers or encoding musical notes into distinguishable (albeit distracting) tactile patterns. There is growing interest in the development of musical haptic systems to augment the auditory experience, although the haptic-music relationship is still not well understood. This paper represents a tactile music interface that provides vibrations to multiple fingertips in synchronicity with auditory music. Like an audio equalizer, different frequency bands are filtered out, and the power in each frequency band is computed and converted to a corresponding vibrational strength. These vibrations are felt on different fingertips, each corresponding to a different frequency band. Songs with music from different spectrums, as classified by their energy and valence, were used to test the effectiveness of the system and to understand the relationship between music and tactile sensations. Three participants were trained on one song categorized as sad (low energy and low valence score) and one song categorized as happy (high energy and high valence score). They were trained both with and without auditory feedback (listening to the song while experiencing the tactile music on their fingertips and then experiencing the vibrations alone without the music). The participants were then tested on three songs from both categories, without any auditory feedback, and were asked to classify the tactile vibrations they felt into either category. The participants were blinded to the songs being tested and were not provided any feedback on the accuracy of their classification. These participants were able to classify the music with 100% accuracy. Although the songs tested were on two opposite spectrums (sad/happy), the preliminary results show the potential of utilizing a vibrotactile equalizer, like the one presented, for augmenting musical experience while furthering the current understanding of music tactile relationship.Keywords: haptic music relationship, tactile equalizer, tactile music, vibrations and mood
Procedia PDF Downloads 16868 Transcriptional Evidence for the Involvement of MyD88 in Flagellin Recognition: Genomic Identification of Rock Bream MyD88 and Comparative Analysis
Authors: N. Umasuthan, S. D. N. K. Bathige, W. S. Thulasitha, I. Whang, J. Lee
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The MyD88 is an evolutionarily conserved host-expressed adaptor protein that is essential for proper TLR/ IL1R immune-response signaling. A previously identified complete cDNA (1626 bp) of OfMyD88 comprised an ORF of 867 bp encoding a protein of 288 amino acids (32.9 kDa). The gDNA (3761 bp) of OfMyD88 revealed a quinquepartite genome organization composed of 5 exons (with the sizes of 310, 132, 178, 92 and 155 bp) separated by 4 introns. All the introns displayed splice signals consistent with the consensus GT/AG rule. A bipartite domain structure with two domains namely death domain (24-103) coded by 1st exon, and TIR domain (151-288) coded by last 3 exons were identified through in silico analysis. Moreover, homology modeling of these two domains revealed a similar quaternary folding nature between human and rock bream homologs. A comprehensive comparison of vertebrate MyD88 genes showed that they possess a 5-exonic structure. In this structure, the last three exons were strongly conserved, and this suggests that a rigid structure has been maintained during vertebrate evolution. A cluster of TATA box-like sequences were found 0.25 kb upstream of cDNA starting position. In addition, putative 5'-flanking region of OfMyD88 was predicted to have TFBS implicated with TLR signaling, including copies of NFB1, APRF/ STAT3, Sp1, IRF1 and 2 and Stat1/2. Using qPCR technique, a ubiquitous mRNA expression was detected in liver and blood. Furthermore, a significantly up-regulated transcriptional expression of OfMyD88 was detected in head kidney (12-24 h; >2-fold), spleen (6 h; 1.5-fold), liver (3 h; 1.9-fold) and intestine (24 h; ~2-fold) post-Fla challenge. These data suggest a crucial role for MyD88 in antibacterial immunity of teleosts.Keywords: MyD88, innate immunity, flagellin, genomic analysis
Procedia PDF Downloads 41167 Altered Gene Expression: Induction/Suppression of some Pathogenesis Related Protein Genes in an Egyptian Isolate of Potato Leafroll Virus (PLRV)
Authors: Dalia G. Aseel
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The potato (Solanum tubersum, L.) has become one of the major vegetable crops in Egypt and all over the world. Potato leafroll virus(PLRV) was observed on potato plants collected from different governorates in Egypt. Three cultivars, Spunta, Diamont, and Cara, infected with PLRV were collected; RNA was extracted and subjected to Real-Time PCR using the coat protein gene primers. The results showed that the expression of the coat protein was 39.6-fold, 12.45-fold, and 47.43-fold, respectively, for Spunta, Diamont, and Cara cultivars. Differential Display Polymerase Chain Reaction (DD-PCR) using pathogenesis-related protein 1 (PR-1), β-1,3-glucanases (PR-2), chitinase (PR-3), peroxidase (POD), and polyphenol oxidase (PPO) forward primers for pathogenesis-related proteins (PR). The obtained data revealed different banding patterns depending on the viral type and the region of infection. Regarding PLRV, 58 up-regulated and 19 down-regulated genes were detected. Sequence analysis of the up-and down-regulated genes revealed that infected plants were observed in comparison with the healthy control. Sequence analysis of the up-regulated gene was performed, and the encoding sequence analysis showed that the obtained genes include: induced stolen tip protein. On the other hand, two down-regulated genes were identified: disease resistance RPP-like protein and non-specific lipid-transfer protein. In this study, the expressions of PR-1, PR-2, PR-3, POD, and PPO genes in the infected leaves of three potato cultivars were estimated by quantitative real-time PCR. We can conclude that the PLRV-infection of potato plants inhibited the expression of the five PR genes. On the contrary, infected leaves by PLRV elevated the expression of some defense genes. This interaction may also induce and/or suppress the expression of some genes responsible for the plant's defense mechanisms.Keywords: PLRV, pathogenesis-related proteins (PRs), DD-PCR, sequence, real-time PCR
Procedia PDF Downloads 13366 Disruption of MoNUC1 Gene Mediates Conidiation in Magnaporthe oryzae
Authors: Irshad Ali Khan, Jian-Ping Lu, Xiao-Hong Liu, Fu-Cheng Lin
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This study reports the functional analysis of a gene MoNUC1 in M. oryzae, which is homologous to the Saccharomyces cerevisiae NUC1 encoding a mitochondrial nuclease protein. The MoNUC1 having a gene locus MGG_05324 is 1002-bp in length and encodes an identical protein of 333 amino acids. We disrupted the gene through gene disruption strategy and isolated two mutants confirmed by southern blotting. The deleted mutants were then used for phenotypic studies and their phenotypes were compared to those of the Guy-11 strain. The mutants were first grown on CM medium to find the effect of MoNUC1 gene disruption on colony growth and the mutants were found to show normal culture colony growth similar to that of the Guy-11 strain. Conidial germination and appressorial formation were also similar in both the mutants and Guy-11 strains showing that this gene plays no significant role in these phenotypes. For pathogenicity, the mutants and Guy-11 mycelium blocks were inoculated on blast susceptible barley seedlings and it was found that both the strains exhibited full pathogenicity showing coalesced and necrotic blast lesions suggesting that this gene is not involved in pathogenicity. Mating of the mutants with 2539 strain formed numerous perithecia showing that MoNUC1 is not essential for sexual reproduction in M. oryzae. However, the mutants were found to form reduced conidia (1.06±8.03B and 1.08±9.80B) than those of the Guy-11 strain (1.46±10.61A) and we conclude that this protein is not required for the blast fungus to cause pathogenicity but plays significant role in conidiation. Proteins of signal transduction pathways that could be disrupted/ intervened genetically or chemically could lead to antifungal products of important fungal cereal diseases and reduce rice yield losses. Tipping the balance toward understanding the whole of pathogenesis, rather than simply conidiation will take some time, but clearly presents the most exciting challenge of all.Keywords: appressorium formation, conidiation, NUC1, Magnaporthe oryzae, pathogenicity
Procedia PDF Downloads 48865 Cytolethal Distending Toxins in Intestinal and Extraintestinal E. coli
Authors: Katarína Čurová, Leonard Siegfried, Radka Vargová, Marta Kmeťová, Vladimír Hrabovský
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Introduction: Cytolethal distending toxins (CDTs) represent intracellular acting proteins which interfere with cell cycle of eukaryotic cells. They are produced by Gram-negative bacteria with afinity to mucocutaneous surfaces and could play a role in the pathogenesis of various diseases. CDTs induce DNA damage probably through DNAse activity, which causes cell cycle arrest and leads to further changes (cell distension and death, apoptosis) depending on the cell type. Five subtypes of CDT (I to V) were reported in E. coli. Methods: We examined 252 E. coli strains belonging to four different groups. Of these strains, 57 were isolated from patients with diarrhea, 65 from patients with urinary tract infections (UTI), 65 from patients with sepsis and 65 from patients with other extraintestinal infections (mostly surgical wounds, decubitus ulcers and respiratory tract infections). Identification of these strains was performed by MALDI-TOF analysis and detection of genes encoding CDTs and determination of the phylogenetic group was performed by PCR. Results: In this study, we detected presence of cdt genes in 11 of 252 E. coli strains tested (4,4 %). Four cdt positive E. coli strains were confirmed in group of UTI (6,15 %), three cdt positive E. coli strains in groups of diarrhea (5,3 %) and other extraintestinal infections (4,6 %). The lowest incidence, one cdt positive E. coli strain, was observed in group of sepsis (1,5 %). All cdt positive E. coli strains belonged to phylogenetic group B2. Conclusion: CDT-producing E. coli are isolated in a low percentage from patients with intestinal and extraintestinal infections, including sepsis and our results correspond with these studies. A weak prevalence of cdt genes suggests that CDTs are not major virulence factors but in combination with other virulence factors may increase virulence potential of E. coli. We suppose that all 11 cdt positive E. coli strains represent real pathogens because they belong to the phylogenetic group B2 which is pathogenic lineage for bacteria E. coli.Keywords: cytolethal distending toxin, E. coli, phylogenetic group, extraintestinal infection, diarrhea
Procedia PDF Downloads 34464 Prevalence of Clostridium perfringens β2-Toxin in Type a Isolates of Sheep and Goats
Authors: Mudassar Mohiuddin, Zahid Iqbal
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Introduction: Clostridium perfringens is an important pathogen responsible for causing enteric diseases in both human and animals. The bacteria produce several toxins. These toxins play vital role in the pathogenesis of various fatal enteric diseases and are classified into five types, on the basis of the differential production of Alpha, Beta, Epsilon and Iota toxins. In addition to the so-called major toxins, there are other toxins like beta2 toxin, produced by some strains of C. perfringens which may play a role in the pathogenesis of disease. Aim of the study: In this study a multiplex PCR assay was developed and used for detection of cpb2 gene to identify the Beta2 harboring isolates among different types of C. perfringens. Objectives: The primary objective of this study was to identify the prevalence of β2-toxin gene in local isolates of Clostridium perfringens. Methodology: This was an experimental study. Random sampling technique was used. A total of 97 sheep and goats were included in this study. All were Pakistani local breeds. The samples were collected during the period from Sep, 2014 to Mar, 2015 from selected districts of Punjab province (Pakistan). Faecal samples were cultured in cooked meat media. The identification of Clostridium perfringens was made on the basis of biochemical tests. Multiplex PCR was performed to identify the toxin genes. Results: A total of 43 C. perfringens isolates were genotyped using multiplex PCR assay. The gene encoding C. perfringens β2-toxin (cpb2) was present in more than 50% of the isolates genotyped. However, the prevalence of this gene varied between sheep and goat isolates. Conclusion: The present study suggests the high occurrence of C. perfringens b2-toxin (cpb2) in the local isolates of Pakistan. As β2-toxin is present in both healthy and diseased animals, so further studies are suggested to establish the role of β2-toxin in pathogenesis of the clostridial enteric diseases.Keywords: beta 2 toxin gene, clostridium perfringens, enteric diseases, goats, multiplex PCR, sheep
Procedia PDF Downloads 45463 Serial Position Curves under Compressively Expanding and Contracting Schedules of Presentation
Authors: Priya Varma, Denis John McKeown
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Psychological time, unlike physical time, is believed to be ‘compressive’ in the sense that the mental representations of a series of events may be internally arranged with ever decreasing inter-event spacing (looking back from the most recently encoded event). If this is true, the record within immediate memory of recent events is severely temporally distorted. Although this notion of temporal distortion of the memory record is captured within some theoretical accounts of human forgetting, notably temporal distinctiveness accounts, the way in which the fundamental nature of the distortion underpins memory and forgetting broadly is barely recognised or at least directly investigated. Our intention here was to manipulate the spacing of items for recall in order to ‘reverse’ this supposed natural compression within the encoding of the items. In Experiment 1 three schedules of presentation (expanding, contracting and fixed irregular temporal spacing) were created using logarithmic spacing of the words for both free and serial recall conditions. The results of recall of lists of 7 words showed statistically significant benefits of temporal isolation, and more excitingly the contracting word series (which we may think of as reversing the natural compression within the mental representation of the word list) showed best performance. Experiment 2 tested for effects of active verbal rehearsal in the recall task; this reduced but did not remove the benefits of our temporal scheduling manipulation. Finally, a third experiment used the same design but with Chinese characters as memoranda, in a further attempt to subvert possible verbal maintenance of items. One change to the design here was to introduce a probe item following the sequence of items and record response times to this probe. Together the outcomes of the experiments broadly support the notion of temporal compression within immediate memory.Keywords: memory, serial position curves, temporal isolation, temporal schedules
Procedia PDF Downloads 20862 Exploring Affordable Care Practs in Nigeria’s Health Insurance Discourse
Authors: Emmanuel Chinaguh, Kehinde Adeosun
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Nigerians die untimely, with 55.75 years of life expectancy, which is 17.45 below the world average of 73.2 (Worldometer, 2020). This is due, among other factors, to the country's limited access to high-quality healthcare. To increase access to good and affordable healthcare services, the National Health Insurance Authority (NHIA) Bill 2022 – which repealed the National Health Insurance Scheme Act 2004 – was passed into law. Applying Jacob Mey’s (2001) pragmatics act (pract) theory, this study explores how NHIA seeks to actualise these healthcare goals by characterising the general situational prototype or pragmemes and pragmatic acts in institutional communications. Data was sourced from the NHIA operational guidelines, which has 147 pages and four sections, and shared posters on NHIA Nigeria Twitter Handle with 14,200 followers. Digital humanities tools, like AntConc and Voyant, were engaged in the data analysis for text encoding and data visualisation. This study identifies these discourse tokens in the data: advertisement and programmes, standards and accreditation, records and information, and offences and penalties. Advertisement and programmes pract facilitating, propagating, prospecting, advising and informing; standards and accreditation, and records and information pract stating, informing and instructing; and offences and penalties pract stating and sanctioning. These practs combined to advance the goals of affordable care and universal accessibility to quality healthcare services. The pragmatic acts were marked by these pragmatic tools: shared situational knowledge (SSK), relevance (REL), reference (REF) and inference (INF). This paper adds to the understanding of health insurance discourse in Nigeria as a mediated social practice that promotes the health of Nigerians.Keywords: affordable care, NHIA, Nigeria’s health insurance discourse, pragmatic acts.
Procedia PDF Downloads 7661 Drought Alters the Expression of a Candidate Zea Mays P-Coumarate 3-Hydroxylase Gene and Caffeic Acid Biosynthesis
Authors: Zintle Kolo, Ndiko Ludidi
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The enzymatic activity of p-coumarate 3-hydroxylase (C3H) synthesize caffeic acid from p-coumaric acid. We recently showed that exogenously applied caffeic acid confers salinity tolerance in soybean (Glycine max) by inducing antioxidant enzymatic activity to promote enhanced scavenging or reactive oxygen species, thus limiting salinity-induced oxidative stress. Recent evidence also establishes that pre-treatment of plants with exogenously supplied caffeic acid improves plant tolerance to osmotic stress by improving plant antioxidant capacity and enhancing biosynthesis of compatible solutes. We aimed to identify a C3H in maize (Zea mays) and evaluate the effect of drought on the spatial and temporal expression of the gene encoding the candidate maize C3H (ZmC3H). Primary sequence analysis shows that ZmC3H shares 71% identity with an Arabidopsis thaliana C3H that is implicated in the control of Arabidopsis cell expansion, growth, and responses to stress. In silico ZmC3H promoter analysis reveals the presence of cis-acting elements that interact with transcription factors implicated in plant responses to drought. Spatial expression analysis by semi-quantitative RT-PCR shows that ZmC3H is expressed in both leaves and roots under normal conditions. However, drought represses the expression of ZmC3H in leaves whereas it up-regulates its expression in roots. These changes in ZmC3H expression correlate with the changes in the content of caffeic acid in maize in response to drought. We illustrate the implications of these changes in the expression of the gene in relation to maize responses to drought and discuss the potential of regulating caffeic acid biosynthesis towards genetic improvement of maize tolerance to drought stress. These findings have implications for food security because of the potential of the implications of the study for drought tolerance in maize.Keywords: caffeic acid, drought-responsive expression, maize drought tolerance, p-coumarate 3-hydroxylase
Procedia PDF Downloads 46360 Lithuanian Sign Language Literature: Metaphors at the Phonological Level
Authors: Anželika Teresė
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In order to solve issues in sign language linguistics, address matters pertaining to maintaining high quality of sign language (SL) translation, contribute to dispelling misconceptions about SL and deaf people, and raise awareness and understanding of the deaf community heritage, this presentation discusses literature in Lithuanian Sign Language (LSL) and inherent metaphors that are created by using the phonological parameter –handshape, location, movement, palm orientation and nonmanual features. The study covered in this presentation is twofold, involving both the micro-level analysis of metaphors in terms of phonological parameters as a sub-lexical feature and the macro-level analysis of the poetic context. Cognitive theories underlie research of metaphors in sign language literature in a range of SL. The study follows this practice. The presentation covers the qualitative analysis of 34 pieces of LSL literature. The analysis employs ELAN software widely used in SL research. The target is to examine how specific types of each phonological parameter are used for the creation of metaphors in LSL literature and what metaphors are created. The results of the study show that LSL literature employs a range of metaphors created by using classifier signs and by modifying the established signs. The study also reveals that LSL literature tends to create reference metaphors indicating status and power. As the study shows, LSL poets metaphorically encode status by encoding another meaning in the same sign, which results in creating double metaphors. The metaphor of identity has been determined. Notably, the poetic context has revealed that the latter metaphor can also be identified as a metaphor for life. The study goes on to note that deaf poets create metaphors related to the importance of various phenomena significance of the lyrical subject. Notably, the study has allowed detecting locations, nonmanual features and etc., never mentioned in previous SL research as used for the creation of metaphors.Keywords: Lithuanian sign language, sign language literature, sign language metaphor, metaphor at the phonological level, cognitive linguistics
Procedia PDF Downloads 13059 Study on Principals Using Change Leadership to Promote School Innovation: A Case Study of a Primary School in Taiwan
Authors: Chih-Wen Fan
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Backgrounds/ Research goals : School improvement requires change leadership, which often means discomfort. Principals are the key people that determine the effectiveness of schools. In an era of organization’s pursuit of speed and effectiveness, school administration has to be accountable and innovative. Effective principals work to improve achievement by focusing on the administrative and teaching quality of improvement. However, there is a lack of literature addressing the relevant case studies on school change leadership. This article explores how principals can use change leadership to drive school change. It analyze the driving factors of principal changes in the case school, the beliefs of change leadership, specific methods, and what impact they have. Methods: This study applies the case study research method to the selected primary school located in an urban area for case study, which has achieved excellent performance after reform and innovation. The researchers selected an older primary school located in an urban area that was transformed into a high-performance primary school after changes were enacted by the principal. The selected case was recommended by three supervisors of the Education Department. The case school underwent leadership change by the new principal during his term, and won an award from the Ministry of Education. Total of 8 teachers are interviewed. The data encoding includes interviews and documents. Expected results/ conclusions: The conclusions of the study are, as follows: (1) The influence for Principal Lin's change leadership is from internal and external environmental development and change pressures. (2) The principal's belief in change leadership is to recognize the sense of crisis, and to create a climate of change and demand for change. (3) The principal's specific actions are intended to identify key members, resolve resistance, use innovative thinking, and promote organizational learning. (4) Principal Lin's change leadership can enhance the professional functions of all employees through appropriate authorization. (5) The effectiveness of change leadership lies in teachers' participation in decision-making; the school's reputation has been enhanced through featured courses.Keywords: change leadership, empowerment, crisis awareness, case study
Procedia PDF Downloads 13358 Dynamic Web-Based 2D Medical Image Visualization and Processing Software
Authors: Abdelhalim. N. Mohammed, Mohammed. Y. Esmail
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In the course of recent decades, medical imaging has been dominated by the use of costly film media for review and archival of medical investigation, however due to developments in networks technologies and common acceptance of a standard digital imaging and communication in medicine (DICOM) another approach in light of World Wide Web was produced. Web technologies successfully used in telemedicine applications, the combination of web technologies together with DICOM used to design a web-based and open source DICOM viewer. The Web server allowance to inquiry and recovery of images and the images viewed/manipulated inside a Web browser without need for any preinstalling software. The dynamic site page for medical images visualization and processing created by using JavaScript and HTML5 advancements. The XAMPP ‘apache server’ is used to create a local web server for testing and deployment of the dynamic site. The web-based viewer connected to multiples devices through local area network (LAN) to distribute the images inside healthcare facilities. The system offers a few focal points over ordinary picture archiving and communication systems (PACS): easy to introduce, maintain and independently platforms that allow images to display and manipulated efficiently, the system also user-friendly and easy to integrate with an existing system that have already been making use of web technologies. The wavelet-based image compression technique on which 2-D discrete wavelet transform used to decompose the image then wavelet coefficients are transmitted by entropy encoding after threshold to decrease transmission time, stockpiling cost and capacity. The performance of compression was estimated by using images quality metrics such as mean square error ‘MSE’, peak signal to noise ratio ‘PSNR’ and compression ratio ‘CR’ that achieved (83.86%) when ‘coif3’ wavelet filter is used.Keywords: DICOM, discrete wavelet transform, PACS, HIS, LAN
Procedia PDF Downloads 15457 Heterogeneity of Genes Encoding the Structural Proteins of Avian Infectious Bronchitis Virus
Authors: Shahid Hussain Abro, Siamak Zohari, Lena H. M. Renström, Désirée S. Jansson, Faruk Otman, Karin Ullman, Claudia Baule
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Infectious bronchitis is an acute, highly contagious respiratory, nephropathogenic and reproductive disease of poultry that is caused by infectious bronchitis virus (IBV). The present study used a large data set of structural gene sequences, including newly generated ones and sequences available in the GenBank database to further analyze the diversity and to identify selective pressures and recombination spots. There were some deletions or insertions in the analyzed regions in isolates of the Italy-02 and D274 genotypes. Whereas, there were no insertions or deletions observed in the isolates of the Massachusetts and 4/91 genotype. The hypervariable nucleotide sequence regions spanned positions 152–239, 554–582, 686–737 and 802–912 in the S1 sub-unit of the all analyzed genotypes. The nucleotide sequence data of the E gene showed that this gene was comparatively unstable and subjected to a high frequency of mutations. The M gene showed substitutions consistently distributed except for a region between nucleotide positions 250–680 that remained conserved. The lowest variation in the nucleotide sequences of ORF5a was observed in the isolates of the D274 genotype. While, ORF5b and N gene sequences showed highly conserved regions and were less subjected to variation. Genes ORF3a, ORF3b, M, ORF5a, ORF5b and N presented negative selective pressure among the analyzed isolates. However, some regions of the ORFs showed favorable selective pressure(s). The S1 and E proteins were subjected to a high rate of mutational substitutions and non-synonymous amino acids. Strong signals of recombination breakpoints and ending break point were observed in the S and N genes. Overall, the results of this study revealed that very likely the strong selective pressures in E, M and the high frequency of substitutions in the S gene can probably be considered the main determinants in the evolution of IBV.Keywords: IBV, avian infectious bronchitis, structural genes, genotypes, genetic diversity
Procedia PDF Downloads 42856 X-Ray Crystallographic Studies on BPSL2418 from Burkholderia pseudomallei
Authors: Mona Alharbi
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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis
Procedia PDF Downloads 23755 Studies on Virulence Factors Analysis in Streptococcus agalactiae from the Clinical Isolates
Authors: Natesan Balasubramanian, Palpandi Pounpandi, Venkatraman Thamil Priya, Vellasamy Shanmugaiah, Karubbiah Balakrishnan, Mandayam Anandam Thirunarayan
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Streptococcus agalactiae is commonly known as Group B Streptococcus (GBS) and it is the most common cause of life-threatening bacterial infection. GBS first considered as a veterinary pathogen causing mastitis in cattle later becomes a human pathogen for severe neonatal infections. In this present study, a total of 20 new clinical isolates of S. agalactiae were collected from male (6) and female patient (14) with different age group. The isolates were from Urinary tract infection (UTI), blood, pus and eye ulcer. All the 20 S. agalactiae isolates has clear hemolysis properties on blood agar medium and were identified by serogrouping and MALTI-TOF-MS analysis. Antibiotic susceptibility/resistance test was performed for 20 S. agalactiae isolates, further phenotypic resistance pattern was observed for tetracycline, vancomycin, ampicillin and penicillin. Genotypically we found two antibiotic resistance genes such as Betalactem antibiotic resistance gene (Tem) (70%) and tetracycline resistance gene Tet(O) 15% in our isolates. Six virulence factors encoding genes were performed by PCR in twenty GBS isolates, cfb gene (100%), followed by, cylE(90.47%), lmp(85.7%), bca(71.42%), rib (38%) and low frequency in bac gene (4.76%) were determined. Most of the S. agalactiae isolates produced strong biofilm in the polystyrene surface (hydrophobic), and low-level biofilm formation was found in glass tube (hydrophilic) surface. lytR is secreted protein and localized in bacterial cell wall, extra cellular membrane, and cytoplasm. In silico docking studies were performed for lytR protein with four antibiofilm compounds, including a peptide (PR39) with the docking study showed peptide has strong interaction followed by ellagic acid and interaction length is 2.95, 2.97 and 2.95 A°. In ligand EGCGO10 and O11 two atoms intract with lytR (Leu271), with binding bond affinity length is 3.24 and 3.14. The aminoacid Leu 271 is act as an impartant aminoacid, since ellagic acid and EGCG interact with same aminoacid.Keywords: antibiotics, biofilms, clinical isolates, S. agalactiae, virulence
Procedia PDF Downloads 10554 Protection and Immune Responses of DNA Vaccines Targeting Virulence Factors of Streptococcus iniae in Nile Tilapia (Oreochromis niloticus)
Authors: Pattanapon Kayansamruaj, Ha Thanh Dong, Nopadon Pirarat, Channarong Rodkhum
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Streptococcus iniae (SI) is a devastating pathogenic bacteria causing heavy mortality in farmed fish. The application of commercialized bacterin vaccine has been reported failures as the outbreaks of the new serotype of SI were emerged in farms after vaccination and subsequently caused severe losses. In the present study, we attempted to develop effective DNA vaccines against SI infection using Nile tilapia (Oreochromis niloticus) as an animal model. Two monovalent DNA vaccines were constructed by the insertion of coding sequences of cell wall-associated virulence factors-encoding genes, comprised of eno (α-enolase) and mtsB (hydrophobic membrane protein), into cytomegalovirus expression vector (pCI-neo). In the animal trial, 30-g Nile tilapia were injected intramuscularly with 15 µg of each vaccine (mock vaccine group was injected by naked pCI-neo) and maintained for 35 days prior challenging with pathogenic SI at the dosage of 107 CFU/fish. At 13 days post-challenge, the relative percent survival of pEno, pMtsB and mock vaccine were 57%, 45% and 27%, respectively. The expression levels of immune responses-associated genes, namely, IL1β, TNF-α, TGF-β, COX2, IL-6, IL-12 and IL-13, were investigated from the spleen of experimental animal at 7 days post-vaccination (PV) and 7 days post-challenge (PC) using quantitative RT-PCR technique. Generally, at 7 days PV, the pEno vaccinated group exhibited highest level of up-regulation (1.7 to 2.9 folds) of every gene, but TGF-β, comparing to pMtsB and mock vaccine groups. However, at 7 days PC, pEno group showed significant up-regulation (1.4 to 8.5 folds) of immune-related genes as similar as mock vaccine group, while pMtsB group had lowest level of up-regulation (0.7 to 3.3 folds). Summarily, this study indicated that the pEno and pMtsB vaccines could elicit the immune responses of the fish and the magnitude of gene expression at 7 days PV was also consistent with the protection level conferred by the vaccine.Keywords: gene expression, DNA vaccine, Nile tilapia, Streptococcus iniae
Procedia PDF Downloads 32453 Targeting the EphA2 Receptor Tyrosine Kinases in Melanoma Cancer, both in Humans and Dogs
Authors: Shabnam Abdi, Behzad Toosi
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Background: Melanoma is the most lethal type of malignant skin cancer in humans and dogs since it spreads rapidly throughout the body. Despite significant advances in treatment, cancer at an advanced stage has a poor prognosis. Hence, more effective treatments are needed to enhance outcomes with fewer side effects. Erythropoietin-producing hepatocellular receptors are the largest family of receptor tyrosine kinases and are divided into two subfamilies, EphA and EphB, both of which play a significant role in disease, especially cancer. Due to their association with proliferation and invasion in many aggressive types of cancer, Eph receptor tyrosine kinases (Eph RTKs) are promising cancer therapy molecules. Because these receptors have not been studied in canine melanoma, we investigated how EphA2 influences survival and tumorigenicity of melanoma cells. Methods: Expression of EphA2 protein in canine melanoma cell lines and human melanoma cell line was evaluated by Western blot. Melanoma cells were transduced with lentiviral particles encoding Eph-targeting shRNAs or non-silencing shRNAs (control) for silencing the expression of EphA2 receptor, and silencing was confirmed by Western blotting and immunofluorescence. The effect of siRNA treatment on cellular proliferation, colony formation, tumorsphere assay, invasion was analyzed by Resazurin assay Matrigel invasion assay, respectively. Results: Expression of EphA2 was detected in canine and human melanoma cell lines. Moreover, stably silencing EphA2 by specific shRNAs significantly and consistently decreased the expression of EphA2 protein in both human and canine melanoma cells. Proliferation, colony formation, tumorsphere and invasion of melanoma cells were significantly decreased in EphA2 siRNA-treated cells compared to control. Conclusion: Our data provide the first functional evidence that the EphA2 receptor plays a critical role in the malignant cellular behavior of melanoma in both human and dogs.Keywords: ephA2, targeting, melanoma, human, canine
Procedia PDF Downloads 5252 A Review Study on the Importance and Correlation of Crisis Literacy and Media Communications for Vulnerable Marginalized People During Crisis
Authors: Maryam Jabeen
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In recent times, there has been a notable surge in attention towards diverse literacy concepts such as media literacy, information literacy, and digital literacy. These concepts have garnered escalating interest, spurring the emergence of novel approaches, particularly in the aftermath of the Covid-19 crisis. However, amidst discussions of crises, the domain of crisis literacy remains largely uncharted within academic exploration. Crisis literacy, also referred to as disaster literacy, denotes an individual's aptitude to not only comprehend but also effectively apply information, enabling well-informed decision-making and adherence to instructions about disaster mitigation, preparedness, response, and recovery. This theoretical and descriptive study seeks to transcend foundational literacy concepts, underscoring the urgency for an in-depth exploration of crisis literacy and its interplay with the realm of media communication. Given the profound impact of the pandemic experience and the looming uncertainty of potential future crises, there arises a pressing need to elevate crisis literacy, or disaster literacy, towards heightened autonomy and active involvement within the spheres of critical disaster preparedness, recovery initiatives, and media communication domains. This research paper is part of my ongoing Ph.D. research study, which explores on a broader level the Encoding and decoding of media communications in relation to crisis literacy. The primary objective of this research paper is to expound upon a descriptive, theoretical research endeavor delving into this domain. The emphasis lies in highlighting the paramount significance of media communications in literacy of crisis, coupled with an accentuated focus on its role in providing information to marginalized populations amidst crises. In conclusion, this research bridges the gap in crisis literacy correlation to media communications exploration, advocating for a comprehensive understanding of its dynamics and its symbiotic relationship with media communications. It intends to foster a heightened sense of crisis literacy, particularly within marginalized communities, catalyzing proactive participation in disaster preparedness, recovery processes, and adept media interactions.Keywords: covid-19, crisis literacy, crisis, marginalized, media and communications, pandemic, vulnerable people
Procedia PDF Downloads 5451 An Attenuated Quadruple Gene Mutant of Mycobacterium tuberculosis Imparts Protection against Tuberculosis in Guinea Pigs
Authors: Shubhita Mathur, Ritika Kar Bahal, Priyanka Chauhan, Anil K. Tyagi
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Mycobacterium tuberculosis, the causative agent of human tuberculosis, is a major cause of mortality. Bacillus Calmette-Guérin (BCG), the only licensed vaccine available for protection against tuberculosis confers highly variable protection ranging from 0%-80%. Thus, novel vaccine strains need to be evaluated for their potential as a vaccine against tuberculosis. We had previously constructed a triple gene mutant of M. tuberculosis (MtbΔmms), having deletions in genes encoding for phosphatases mptpA, mptpB, and sapM that are involved in host-pathogen interaction. Though vaccination with Mtb∆mms strain induced protection in the lungs of guinea pigs, the mutant strain was not able to control the hematogenous spread of the challenge strain to the spleens. Additionally, inoculation with Mtb∆mms resulted in some pathological damage to the spleens in the early phase of infection. In order to overcome the pathology caused by MtbΔmms in the spleens of guinea pigs and also to control the dissemination of the challenge strain, MtbΔmms was genetically modified by disrupting bioA gene to generate MtbΔmmsb strain. Further, in vivo attenuation of MtbΔmmsb was evaluated, and its protective efficacy was assessed against virulent M. tuberculosis challenge in guinea pigs. Our study demonstrates that Mtb∆mmsb mutant was highly attenuated for growth and virulence in guinea pigs. Vaccination with Mtb∆mmsb mutant generated significant protection in comparison to sham-immunized animals at 4 and 12 weeks post-infection in lungs and spleens of the infected animals. Our findings provide evidence that deletion of genes involved in signal transduction and biotin biosynthesis severely attenuates the pathogen and the single immunization with the auxotroph was able to provide significant protection as compared to sham-immunized animals. The protection imparted by Mtb∆mmsb fell short in comparison to the protection observed in BCG-immunized animals. This study nevertheless indicates the importance of attenuated multiple gene deletion mutants of M. tuberculosis in generating protection against tuberculosis.Keywords: Mycobacterium tuberculosis, BCG, MtbΔmmsb, bioA, guinea pigs
Procedia PDF Downloads 12550 Creating and Questioning Research-Oriented Digital Outputs to Manuscript Metadata: A Case-Based Methodological Investigation
Authors: Diandra Cristache
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The transition of traditional manuscript studies into the digital framework closely affects the methodological premises upon which manuscript descriptions are modeled, created, and questioned for the purpose of research. This paper intends to explore the issue by presenting a methodological investigation into the process of modeling, creating, and questioning manuscript metadata. The investigation is founded on a close observation of the Polonsky Greek Manuscripts Project, a collaboration between the Universities of Cambridge and Heidelberg. More than just providing a realistic ground for methodological exploration, along with a complete metadata set for computational demonstration, the case study also contributes to a broader purpose: outlining general methodological principles for making the most out of manuscript metadata by means of research-oriented digital outputs. The analysis mainly focuses on the scholarly approach to manuscript descriptions, in the specific instance where the act of metadata recording does not have a programmatic research purpose. Close attention is paid to the encounter of 'traditional' practices in manuscript studies with the formal constraints of the digital framework: does the shift in practices (especially from the straight narrative of free writing towards the hierarchical constraints of the TEI encoding model) impact the structure of metadata and its capability to respond specific research questions? It is argued that flexible structure of TEI and traditional approaches to manuscript description lead to a proliferation of markup: does an 'encyclopedic' descriptive approach ensure the epistemological relevance of the digital outputs to metadata? To provide further insight on the computational approach to manuscript metadata, the metadata of the Polonsky project are processed with techniques of distant reading and data networking, thus resulting in a new group of digital outputs (relational graphs, geographic maps). The computational process and the digital outputs are thoroughly illustrated and discussed. Eventually, a retrospective analysis evaluates how the digital outputs respond to the scientific expectations of research, and the other way round, how the requirements of research questions feed back into the creation and enrichment of metadata in an iterative loop.Keywords: digital manuscript studies, digital outputs to manuscripts metadata, metadata interoperability, methodological issues
Procedia PDF Downloads 13549 Wide Dissemination of CTX-M-Type Extended-Spectrum β-Lactamases in Korean Swine Farms
Authors: Young Ah Kim, Hyunsoo Kim, Eun-Jeong Yoon, Young Hee Seo, Kyungwon Lee
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Extended-spectrum β-lactamase (ESBL)-producing Escherichia coli from food animals are considered as a reservoir for transmission of ESBL genes to human. The aim of this study is to assess the prevalence and molecular epidemiology of ESBL-producing E. coli colonization in pigs, farm workers, and farm environments to elucidate the transmission of multidrug-resistant clones from animal to human. Nineteen pig farms were enrolled across the country in Korea from August to December 2017. ESBL-producing E. coli isolates were detected in 190 pigs, 38 farm workers, and 112 sites of farm environments using ChromID ESBL (bioMerieux, Marcy l'Etoile, France), directly (stool or perirectal swab) or after enrichment (sewage). Antimicrobial susceptibility tests were done with disk diffusion methods and blaTEM, blaSHV, and blaCTX-M were detected with PCR and sequencing. The genomes of the four CTX-M-55-producing E. coli isolates from various sources in one farm were entirely sequenced to assess the relatedness of the strains. Whole genome sequencing (WGS) was performed with PacBio RS II system (Pacific Biosciences, Menlo Park, CA, USA). ESBL genotypes were 85 CTX-M-1 group (one CTX-M-3, 23 CTX-M-15, one CTX-M-28, 59 CTX-M-55, one CTX-M-69) and 60 CTX-M-9 group (41 CTX-M-14, one CTX-M-17, one CTX-M-27, 13 CTX-M-65, 4 CTX-M-102) in total 145 isolates. The rectal colonization rates were 53.2% (101/190) in pigs and 39.5% (15/38) in farm workers. In WGS, sequence types (STs) were determined as ST69 (E. coli PJFH115 isolate from a human carrier), ST457 (two E. coli isolates PJFE101 recovered from a fence and PJFA1104 from a pig) and ST5899 (E. coli PJFA173 isolate from the other pig). The four plasmids encoding CTX-M-55 (88,456 to 149, 674 base pair), whether it belonged to IncFIB or IncFIC-IncFIB type, shared IncF backbone furnishing the conjugal elements, suggesting of genes originated from same ancestor. In conclusion, the prevalence of ESBL-producing E. coli in swine farms was surprisingly high, and many of them shared common ESBL genotypes of clinical isolates such as CTX-M-14, 15, and 55 in Korea. It could spread by horizontal transfer between isolates from different reservoirs (human-animal-environment).Keywords: Escherichia coli, extended-spectrum β-lactamase, prevalence, whole genome sequencing
Procedia PDF Downloads 19948 Autosomal Dominant Polycystic Kidney Patients May Be Predisposed to Various Cardiomyopathies
Authors: Fouad Chebib, Marie Hogan, Ziad El-Zoghby, Maria Irazabal, Sarah Senum, Christina Heyer, Charles Madsen, Emilie Cornec-Le Gall, Atta Behfar, Barbara Ehrlich, Peter Harris, Vicente Torres
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Background: Mutations in PKD1 and PKD2, the genes encoding the proteins polycystin-1 (PC1) and polycystin-2 (PC2) cause autosomal dominant polycystic kidney disease (ADPKD). ADPKD is a systemic disease associated with several extrarenal manifestations. Animal models have suggested an important role for the polycystins in cardiovascular function. The aim of the current study is to evaluate the association of various cardiomyopathies in a large cohort of patients with ADPKD. Methods: Clinical data was retrieved from medical records for all patients with ADPKD and cardiomyopathies (n=159). Genetic analysis was performed on available DNA by direct sequencing. Results: Among the 58 patients included in this case series, 39 patients had idiopathic dilated cardiomyopathy (IDCM), 17 had hypertrophic obstructive cardiomyopathy (HOCM), and 2 had left ventricular noncompaction (LVNC). The mean age at cardiomyopathy diagnosis was 53.3, 59.9 and 53.5 years in IDCM, HOCM and LVNC patients respectively. The median left ventricular ejection fraction at initial diagnosis of IDCM was 25%. Average basal septal thickness was 19.9 mm in patients with HOCM. Genetic data was available in 19, 8 and 2 cases of IDCM, HOCM, and LVNC respectively. PKD1 mutations were detected in 47.4%, 62.5% and 100% of IDCM, HOCM and LVNC cases. PKD2 mutations were detected only in IDCM cases and were overrepresented (36.8%) relative to the expected frequency in ADPKD (~15%). The prevalence of IDCM, HOCM, and LVNC in our ADPKD clinical cohort was 1:17, 1:39 and 1:333 respectively. When compared to the general population, IDCM and HOCM was approximately 10-fold more prevalent in patients with ADPKD. Conclusions: In summary, we suggest that PKD1 or PKD2 mutations may predispose to idiopathic dilated or hypertrophic cardiomyopathy. There is a trend for patients with PKD2 mutations to develop the former and for patients with PKD1 mutations to develop the latter. Predisposition to various cardiomyopathies may be another extrarenal manifestation of ADPKD.Keywords: autosomal dominant polycystic kidney (ADPKD), polycystic kidney disease, cardiovascular, cardiomyopathy, idiopathic dilated cardiomyopathy, hypertrophic cardiomyopathy, left ventricular noncompaction
Procedia PDF Downloads 30347 Characterization of a Putative Type 1 Toxin-Antitoxin System in Shigella Flexneri
Authors: David Sarpong, Waleed Khursheed, Ernest Danquah, Erin Murphy
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Shigella is a pathogenic bacterium responsible for shigellosis, a severe diarrheal disease that claims the lives of immunocompromised individuals worldwide. To develop therapeutics against this disease, an understanding of the molecular mechanisms underlying the pathogen’s physiology is crucial. Small non-coding RNAs (sRNAs) have emerged as important regulators of bacterial physiology, including as components of toxin-antitoxin systems. In this study, we investigated the role of RyfA in S. flexneri physiology and virulence. RyfA, originally identified as an sRNA in Escherichia coli, is conserved within the Enterobacteriaceae family, including Shigella. Whereas two copies of ryfA are present in S. dysenteriae, all other Shigella species contain only one copy of the gene. Additionally, we identified a putative open reading frame within the RyfA transcript, suggesting that it may be a dual-functioning gene encoding a small protein in addition to its sRNA function. To study ryfA in vitro, we cloned the gene into an inducible plasmid and observed the effect on bacterial growth. Here, we report that RyfA production inhibits the growth of S. flexneri, and this inhibition is dependent on the contained open reading frame. In-silico analyses have revealed the presence of two divergently transcribed sRNAs, RyfB1 and RyfB2, which share nucleotide complementarity with RyfA and thus are predicted to function as anti-toxins. Our data demonstrate that RyfB2 has a stronger antitoxin effect than RyfB1. This regulatory pattern suggests a novel form of a toxin-antitoxin system in which the activity of a single toxin is inhibited to varying degrees by two sRNA antitoxins. Studies are ongoing to investigate the regulatory mechanism(s) of the antitoxin genes, as well as the downstream targets and mechanism of growth inhibition by the RyfA toxin. This study offers distinct insights into the regulatory mechanisms underlying Shigella physiology and may inform the development of new anti-Shigella therapeutics.Keywords: sRNA, shigella, toxin-antitoxin, Type 1 toxin antitoxin
Procedia PDF Downloads 3946 Comparison of Existing Predictor and Development of Computational Method for S- Palmitoylation Site Identification in Arabidopsis Thaliana
Authors: Ayesha Sanjana Kawser Parsha
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S-acylation is an irreversible bond in which cysteine residues are linked to fatty acids palmitate (74%) or stearate (22%), either at the COOH or NH2 terminal, via a thioester linkage. There are several experimental methods that can be used to identify the S-palmitoylation site; however, since they require a lot of time, computational methods are becoming increasingly necessary. There aren't many predictors, however, that can locate S- palmitoylation sites in Arabidopsis Thaliana with sufficient accuracy. This research is based on the importance of building a better prediction tool. To identify the type of machine learning algorithm that predicts this site more accurately for the experimental dataset, several prediction tools were examined in this research, including the GPS PALM 6.0, pCysMod, GPS LIPID 1.0, CSS PALM 4.0, and NBA PALM. These analyses were conducted by constructing the receiver operating characteristics plot and the area under the curve score. An AI-driven deep learning-based prediction tool has been developed utilizing the analysis and three sequence-based input data, such as the amino acid composition, binary encoding profile, and autocorrelation features. The model was developed using five layers, two activation functions, associated parameters, and hyperparameters. The model was built using various combinations of features, and after training and validation, it performed better when all the features were present while using the experimental dataset for 8 and 10-fold cross-validations. While testing the model with unseen and new data, such as the GPS PALM 6.0 plant and pCysMod mouse, the model performed better, and the area under the curve score was near 1. It can be demonstrated that this model outperforms the prior tools in predicting the S- palmitoylation site in the experimental data set by comparing the area under curve score of 10-fold cross-validation of the new model with the established tools' area under curve score with their respective training sets. The objective of this study is to develop a prediction tool for Arabidopsis Thaliana that is more accurate than current tools, as measured by the area under the curve score. Plant food production and immunological treatment targets can both be managed by utilizing this method to forecast S- palmitoylation sites.Keywords: S- palmitoylation, ROC PLOT, area under the curve, cross- validation score
Procedia PDF Downloads 6745 Encoding the Design of the Memorial Park and the Family Network as the Icon of 9/11 in Amy Waldman's the Submission
Authors: Masami Usui
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After 9/11, the American literary scene was confronted with new perspectives that enabled both writers and readers to recognize the hidden aspects of their political, economic, legal, social, and cultural phenomena. There appeared an argument over new and challenging multicultural aspects after 9/11 and this argument is presented by a tension of space related to 9/11. In Amy Waldman’s the Submission (2011), designing both the memorial park and the family network has a significant meaning in establishing the progress of understanding from multiple perspectives. The most intriguing and controversial topic of racism is reflected in the Submission, where one young architect’s blind entry to the competition for the memorial of Ground Zero is nominated, yet he is confronted with strong objections and hostility as soon as he turns out to be a Muslim named Mohammad Khan. This ‘Khan’ issue, immediately enlarged into a social controversial issue on American soil, causes repeated acts of hostility to Muslim women by ignorant citizens all over America. His idea of the park is to design a new concept of tracing the cultural background of the open space. Against his will, his name is identified as the ‘ingredient’ of the networking of the resistant community with his supporters: on the other hand, the post 9/11 hysteria and victimization is presented in such family associations as the Angry Family Members and Grieving Family Members. These rapidly expanding networks, whether political or not, constructed by the internet, embody the contemporary societal connection and representation. The contemporary quest for the significance of human relationships is recognized as a quest for global peace. Designing both the memorial park and the communication networks strengthens a process of facing the shared conflicts and healing the survivors’ trauma. The tension between the idea and networking of the Garden for the memorial site and the collapse of Ground Zero signifies the double mission of the site: to establish the space to ease the wounded and to remember the catastrophe. Reading the design of these icons of 9/11 in the Submission means that decoding the myth of globalization and its representations in this century.Keywords: American literature, cultural studies, globalization, literature of catastrophe
Procedia PDF Downloads 52244 Expression of Human Papillomavirus Type 18 L1 Virus-Like Particles in Methylotropic Yeast, Pichia Pastoris
Authors: Hossein Rassi, Marjan Moradi Fard, Samaneh Niko
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Human papillomavirus type 16 and 18 are closely associated with the development of human cervical carcinoma, which is one of the most common causes of cancer death in women worldwide. At present, HPV type 18 accounts for about 34 % of all HPV infections in Iran and the most promising vaccine against HPV infection is based on the L1 major capsid protein. The L1 protein of HPV18 has the capacity to self-assemble into capsomers or virus-like particles (VLPs) that are non-infectious, highly immunogenic and allowing their use in vaccine production. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system used to produce high levels of heterologous proteins. In this study we expressed HPV18 L1 VLPs in P. pastoris. The gene encoding the major capsid protein L1 of the high-risk HPV type 18 was isolated from Iranian patient by PCR and inserted into pTG19-T vector to obtain the recombinant expression vector pTG19-HPV18-L1. Then, the pTG19-HPV18-L1 was transformed into E. coli strain DH5α and the recombinant protein HPV18 L1 was expressed under IPTG induction in soluble form. The HPV18 L1 gene was excised from recombinant plasmid with XhoI and EcoRI enzymes and ligated into the yeast expression vector pPICZα linearized with the same enzymes, and transformed into P. pastoris. Induction and expression of HPV18 L1 protein was demonstrated by BMGY/BMMY and RT PCR. The parameters for induced cultivation for strain in P. pastoris KM71 with HPV16L1 were investigated in shaking flask cultures. After induced cultivation BMMY (pH 7.0) medium supplemented with methanol to a final concentration of 1.0% every 24 h at 37 degrees C for 96 h, the recombinant produced 78.6 mg/L of L1 protein. This work offers the possibility for the production of prophylactic vaccine for cervical carcinoma by P. pastoris for HPV-18 L1 gene. The VLP-based HPV vaccines can prevent persistent HPV18 infections and cervical cancer in Iran. The HPV-18 L1 gene was expressed successfully in E.coli, which provides necessary basis for preparing HPV-18 L1 vaccine in human. Also, HPV type 6 L1 proteins expressed in Pichia pastoris will facilitate the HPV vaccine development and structure-function study.Keywords: Pichia pastoris, L1 virus-like particles, human papillomavirus type 18, biotechnology
Procedia PDF Downloads 40143 Update on Epithelial Ovarian Cancer (EOC), Types, Origin, Molecular Pathogenesis, and Biomarkers
Authors: Salina Yahya Saddick
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Ovarian cancer remains the most lethal gynecological malignancy due to the lack of highly sensitive and specific screening tools for detection of early-stage disease. The OSE provides the progenitor cells for 90% of human ovarian cancers. Recent morphologic, immunohistochemical and molecular genetic studies have led to the development of a new paradigm for the pathogenesis and origin of epithelial ovarian cancer (EOC) based on a ualistic model of carcinogenesis that divides EOC into two broad categories designated Types I and II which are characterized by specific mutations, including KRAS, BRAF, ERBB2, CTNNB1, PTEN PIK3CA, ARID1A, and PPPR1A, which target specific cell signaling pathways. Type 1 tumors rarely harbor TP53. type I tumors are relatively genetically stable and typically display a variety of somatic sequence mutations that include KRAS, BRAF, PTEN, PIK3CA CTNNB1 (the gene encoding beta catenin), ARID1A and PPP2R1A but very rarely TP53 . The cancer stem cell (CSC) hypothesis postulates that the tumorigenic potential of CSCs is confined to a very small subset of tumor cells and is defined by their ability to self-renew and differentiate leading to the formation of a tumor mass. Potential protein biomarker miRNA, are promising biomarkers as they are remarkably stable to allow isolation and analysis from tissues and from blood in which they can be found as free circulating nucleic acids and in mononuclear cells. Recently, genomic anaylsis have identified biomarkers and potential therapeutic targets for ovarian cancer namely, FGF18 which plays an active role in controlling migration, invasion, and tumorigenicity of ovarian cancer cells through NF-κB activation, which increased the production of oncogenic cytokines and chemokines. This review summarizes update information on epithelial ovarian cancers and point out to the most recent ongoing research.Keywords: epithelial ovarian cancers, somatic sequence mutations, cancer stem cell (CSC), potential protein, biomarker, genomic analysis, FGF18 biomarker
Procedia PDF Downloads 37242 A Web-Based Systems Immunology Toolkit Allowing the Visualization and Comparative Analysis of Publically Available Collective Data to Decipher Immune Regulation in Early Life
Authors: Mahbuba Rahman, Sabri Boughorbel, Scott Presnell, Charlie Quinn, Darawan Rinchai, Damien Chaussabel, Nico Marr
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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 28941 Transcriptomic Analysis for Differential Expression of Genes Involved in Secondary Metabolite Production in Narcissus Bulb and in vitro Callus
Authors: Aleya Ferdausi, Meriel Jones, Anthony Halls
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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 14040 Constitutive Flo1p Expression on Strains Bearing Deletions in Genes Involved in Cell Wall Biogenesis
Authors: Lethukuthula Ngobese, Abin Gupthar, Patrick Govender
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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 407