Search results for: sequencing skill
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1296

Search results for: sequencing skill

126 Validation of Global Ratings in Clinical Performance Assessment

Authors: S. J. Yune, S. Y. Lee, S. J. Im, B. S. Kam, S. Y. Baek

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This study aimed to determine the reliability of clinical performance assessments, having been emphasized by ability-based education, and professors overall assessment methods. We addressed the following problems: First, we try to find out whether there is a difference in what we consider to be the main variables affecting the clinical performance test according to the evaluator’s working period and the number of evaluation experience. Second, we examined the relationship among the global rating score (G), analytic global rating score (Gc), and the sum of the analytical checklists (C). What are the main factors affecting clinical performance assessments in relation to the numbers of times the evaluator had administered evaluations and the length of their working period service? What is the relationship between overall assessment score and analytic checklist score? How does analytic global rating with 6 components in OSCE and 4 components in sub-domains (Gc) CPX: aseptic practice, precision, systemic approach, proficiency, successfulness, and attitude overall assessment score and task-specific analytic checklist score sum (C) affect the professor’s overall global rating assessment score (G)? We studied 75 professors who attended a 2016 Bugyeoung Consortium clinical skills performances test evaluating third and fourth year medical students at the Pusan National University Medical school in South Korea (39 prof. in OSCE, 36 prof. in CPX; all consented to participate in our study). Each evaluator used 3 forms; a task-specific analytic checklist, subsequent analytic global rating scale with sub-6 domains, and overall global scale. After the evaluation, the professors responded to the questionnaire on the important factors of clinical performance assessment. The data were analyzed by frequency analysis, correlation analysis, and hierarchical regression analysis using SPSS 21.0. Their understanding of overall assessment was analyzed by dividing the subjects into groups based on experiences. As a result, they considered ‘precision’ most important in overall OSCE assessment, and ‘precise accuracy physical examination’, ‘systemic approaches to taking patient history’, and ‘diagnostic skill capability’ in overall CPX assessment. For OSCE, there was no clear difference of opinion about the main factors, but there was for CPX. Analytic global rating scale score, overall rating scale score, and analytic checklist score had meaningful mutual correlations. According to the regression analysis results, task-specific checklist score sum had the greatest effect on overall global rating. professors regarded task-specific analytic checklist total score sum as best reflecting overall OSCE test score, followed by aseptic practice, precision, systemic approach, proficiency, successfulness, and attitude on a subsequent analytic global rating scale. For CPX, subsequent analytic global rating scale score, overall global rating scale score, and task-specific checklist score had meaningful mutual correlations. These findings support explanations for validity of professors’ global rating in clinical performance assessment.

Keywords: global rating, clinical performance assessment, medical education, analytic checklist

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125 Integrative Omics-Portrayal Disentangles Molecular Heterogeneity and Progression Mechanisms of Cancer

Authors: Binder Hans

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Cancer is no longer seen as solely a genetic disease where genetic defects such as mutations and copy number variations affect gene regulation and eventually lead to aberrant cell functioning which can be monitored by transcriptome analysis. It has become obvious that epigenetic alterations represent a further important layer of (de-)regulation of gene activity. For example, aberrant DNA methylation is a hallmark of many cancer types, and methylation patterns were successfully used to subtype cancer heterogeneity. Hence, unraveling the interplay between different omics levels such as genome, transcriptome and epigenome is inevitable for a mechanistic understanding of molecular deregulation causing complex diseases such as cancer. This objective requires powerful downstream integrative bioinformatics methods as an essential prerequisite to discover the whole genome mutational, transcriptome and epigenome landscapes of cancer specimen and to discover cancer genesis, progression and heterogeneity. Basic challenges and tasks arise ‘beyond sequencing’ because of the big size of the data, their complexity, the need to search for hidden structures in the data, for knowledge mining to discover biological function and also systems biology conceptual models to deduce developmental interrelations between different cancer states. These tasks are tightly related to cancer biology as an (epi-)genetic disease giving rise to aberrant genomic regulation under micro-environmental control and clonal evolution which leads to heterogeneous cellular states. Machine learning algorithms such as self organizing maps (SOM) represent one interesting option to tackle these bioinformatics tasks. The SOMmethod enables recognizing complex patterns in large-scale data generated by highthroughput omics technologies. It portrays molecular phenotypes by generating individualized, easy to interpret images of the data landscape in combination with comprehensive analysis options. Our image-based, reductionist machine learning methods provide one interesting perspective how to deal with massive data in the discovery of complex diseases, gliomas, melanomas and colon cancer on molecular level. As an important new challenge, we address the combined portrayal of different omics data such as genome-wide genomic, transcriptomic and methylomic ones. The integrative-omics portrayal approach is based on the joint training of the data and it provides separate personalized data portraits for each patient and data type which can be analyzed by visual inspection as one option. The new method enables an integrative genome-wide view on the omics data types and the underlying regulatory modes. It is applied to high and low-grade gliomas and to melanomas where it disentangles transversal and longitudinal molecular heterogeneity in terms of distinct molecular subtypes and progression paths with prognostic impact.

Keywords: integrative bioinformatics, machine learning, molecular mechanisms of cancer, gliomas and melanomas

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124 Teacher Characteristics That Influence Development of Oral Language Skills among Pre-Primary School Pupils: Case Study of Nairobi City County, Kenya

Authors: Kenneth Okelo, Esther Waithaka, Maureen Mweru

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Development of oral language skills is a precursor to writing and reading acquisition. Oral skill is a means of communication through which people express their desires, ideas, excitements, amusements, disappointments and exchange information. In addition, oral skills have been found to be an important tool for thinking and concept development in children. Research carried out in industrialised countries have identified some appropriate teaching strategies used to enhance acquisition of oral language skills such as repetition, substitution, explanation, contrast, exemplification and code-switching. However, these studies’ geographical locations do not reflect the diversity of the Kenyan society. In addition, studies conducted in Kenya in the past have not established why pre-primary school teachers are not using appropriate teaching strategies. The purpose of this study was to find out whether teachers’ experience, academic qualification and type of training influences their choice of teaching strategies in the development of oral language skills inside and out of the classroom in selected preschools in Kibra Sub-County, Nairobi County. In addition, this study aimed at finding out the strategies used by teachers in Kibra Sub-County to promote oral skills development among pre-primary school children. The study was guided by Holdaway’s theory of language acquisition. Descriptive survey design was employed during this study. Questionnaires and observation schedules were used to collect data. Eighty-three (83) preschool teachers were sampled using multistage sampling methods for observation. Data was analysed using SPSS version 20. The researcher carried out content analysis on the qualitative data. The main descriptive methods used were tabulation of frequencies and percentages. Chi squire test was the inferential statistic used to test the relationship between variables. The main findings of the study indicate that teaching strategies that were mostly used by pre-primary school teachers were code-switching, examples, repetition, substitution and explanation. While questions, direction, expansion of children words and contrast were the least used teaching strategies when teaching oral language skills. The study revealed that the there is a slight correlation between the type of training of teachers and the teaching strategies as most of DICECE trained teachers used more teaching strategies when teaching oral skills compared to other teachers. The findings also revealed that there was a partial significant correlation between teacher’s academic qualifications and a few teaching strategies. A similar correlation was also observed between teaching experience and a few teaching strategies. Since the strategies used by pre-primary school teachers under the study were less than half of the recommended teaching strategies to promote oral skills, the study recommends that teachers should be encouraged to use more in structural strategies to improve children’s oral language skills.

Keywords: Kenya early childhood education, Kenya education, oral language skills acquisition, teaching methods

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123 Teamwork on Innovation in Young Enterprises: A Qualitative Analysis

Authors: Polina Trusova

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The majority of young enterprises is founded and run by teams and develops new, innovative products or services. While problems within the team are considered to be an important reason for the failure of young enterprises, effective teamwork on innovation may be a key success factor. It may require special teamwork design or members’ creativity not needed during work routine. However, little is known about how young enterprises develop innovative solutions in teams, what makes their teamwork special and what influences its effectivity. Extending this knowledge is essential for understanding the success and failure factors for young enterprises. Previous research focused on working on innovation or professional teams in general. Rare studies combining these issues usually concentrate on homogenous groups like IT expert teams in innovation projects of big, well-established firms. The transferability of those studies’ findings to the entrepreneurial context is doubtful because of several reasons why teamwork should differ significantly between big, well-established firms and young enterprises. First, teamwork is conducted by team members, e.g., employees. The personality of employees in young enterprises, in contrast to that of employees in established firms, has been shown to be more similar to the personality of entrepreneurs. As entrepreneurs were found to be more open to experience and show less risk aversion, it may have a positive impact on their teamwork. Persons open to novelty are more likely to develop or accept a creative solution, which is especially important for teamwork on innovation. Secondly, young enterprises are often characterized by a flat hierarchy, so in general, teamwork should be more participative there. It encourages each member (and not only the founder) to produce and discuss innovative ideas, increasing their variety and enabling the team to select the best idea from the larger idea pool. Thirdly, teams in young enterprises are often multidisciplinary. It has some advantages but also increases the risk of internal conflicts making teamwork less effective. Despite the key role of teamwork on innovation and presented barriers for transferring existing evidence to the context of young enterprises, only a few researchers have addressed this issue. In order to close the existing research gap, to explore and understand how innovations are developed in teams of young enterprises and which factors influencing teamwork may be especially relevant for such teams, a qualitative study has been developed. The study consisting of 20 half-structured interviews with (co-)founders of young innovative enterprises in the UK and USA started in September 2017. The interview guide comprises but is not limited to teamwork dimensions discussed in literature like members’ skill or authority differentiation. Data will be evaluated following the rules of qualitative content analysis. First results indicate some factors which may be relevant especially for teamwork in young innovative enterprises. They will enrich the scientific discussion and provide the evidence needed to test a possible causality between identified factors and teamwork effectivity in future research on young innovative enterprises. Results and their discussion can be presented at the conference.

Keywords: innovation, qualitative study, teamwork, young enterprises

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122 Exploring Fluoroquinolone-Resistance Dynamics Using a Distinct in Vitro Fermentation Chicken Caeca Model

Authors: Bello Gonzalez T. D. J., Setten Van M., Essen Van A., Brouwer M., Veldman K. T.

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Resistance to fluoroquinolones (FQ) has evolved increasingly over the years, posing a significant challenge for the treatment of human infections, particularly gastrointestinal tract infections caused by zoonotic bacteria transmitted through the food chain and environment. In broiler chickens, a relatively high proportion of FQ resistance has been observed in Escherichia coli indicator, Salmonella and Campylobacter isolates. We hypothesize that flumequine (Flu), used as a secondary choice for the treatment of poultry infections, could potentially be associated with a high proportion of FQ resistance. To evaluate this hypothesis, we used an in vitro fermentation chicken caeca model. Two continuous single-stage fermenters were used to simulate in real time the physiological conditions of the chicken caeca microbial content (temperature, pH, caecal content mixing, and anoxic environment). A pool of chicken caecal content containing FQ-resistant E. coli obtained from chickens at slaughter age was used as inoculum along with a spiked FQ-susceptible Campylobacter jejuni strain isolated from broilers. Flu was added to one of the fermenters (Flu-fermenter) every 24 hours for two days to evaluate the selection and maintenance of FQ resistance over time, while the other served as a control (C-Fermenter). The experiment duration was 5 days. Samples were collected at three different time points: before, during and after Flu administration. Serial dilutions were plated on Butzler culture media with and without Flu (8mg/L) and enrofloxacin (4mg/L) and on MacConkey culture media with and without Flu (4mg/L) and enrofloxacin (1mg/L) to determine the proportion of resistant strains over time. Positive cultures were identified by mass spectrometry and matrix-assisted laser desorption/ionization (MALDI). A subset of the obtained isolates were used for Whole Genome Sequencing analysis. Over time, E. coli exhibited positive growth in both fermenters, while C. jejuni growth was detected up to day 3. The proportion of Flu-resistant E. coli strains recovered remained consistent over time after antibiotic selective pressure, while in the C-fermenter, a decrease was observed at day 5; a similar pattern was observed in the enrofloxacin-resistant E. coli strains. This suggests that Flu might play a role in the selection and persistence of enrofloxacin resistance, compared to C-fermenter, where enrofloxacin-resistant E. coli strains appear at a later time. Furthermore, positive growth was detected from both fermenters only on Butzler plates without antibiotics. A subset of C. jejuni strains from the Flu-fermenter revealed that those strains were susceptible to ciprofloxacin (MIC < 0.12 μg/mL). A selection of E. coli strains from both fermenters revealed the presence of plasmid-mediated quinolone resistance (PMQR) (qnr-B19) in only one strain from the C-fermenter belonging to sequence type (ST) 48, and in all from Flu-fermenter belonged to ST189. Our results showed that Flu selective impact on PMQR-positive E. coli strains, while no effect was observed in C. jejuni. Maintenance of Flu-resistance was correlated with antibiotic selective pressure. Further studies into antibiotic resistance gene transfer among commensal and zoonotic bacteria in the chicken caeca content may help to elucidate the resistance spread mechanisms.

Keywords: fluoroquinolone-resistance, escherichia coli, campylobacter jejuni, in vitro model

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121 Bacterial Community Diversity in Soil under Two Tillage Systems

Authors: Dalia Ambrazaitienė, Monika Vilkienė, Danute Karcauskienė, Gintaras Siaudinis

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The soil is a complex ecosystem that is part of our biosphere. The ability of soil to provide ecosystem services is dependent on microbial diversity. T Tillage is one of the major factors that affect soil properties. The no-till systems or shallow ploughless tillage are opposite of traditional deep ploughing, no-tillage systems, for instance, increase soil organic matter by reducing mineralization rates and stimulating litter concentrations of the top soil layer, whereas deep ploughing increases the biological activity of arable soil layer and reduces the incidence of weeds. The role of soil organisms is central to soil processes. Although the number of microbial species in soil is still being debated, the metagenomic approach to estimate microbial diversity predicted about 2000 – 18 000 bacterial genomes in 1 g of soil. Despite the key role of bacteria in soil processes, there is still lack of information about the bacterial diversity of soils as affected by tillage practices. This study focused on metagenomic analysis of bacterial diversity in long-term experimental plots of Dystric Epihypogleyic Albeluvisols in western part of Lithuania. The experiment was set up in 2013 and had a split-plot design where the whole-plot treatments were laid out in a randomized design with three replicates. The whole-plot treatments consisted of two tillage methods - deep ploughing (22-25 cm) (DP), ploughless tillage (7-10 cm) (PT). Three subsamples (0-20 cm) were collected on October 22, 2015 for each of the three replicates. Subsamples from the DP and PT systems were pooled together wise to make two composition samples, one representing deep ploughing (DP) and the other ploughless tillage (PT). Genomic DNA from soil sample was extracted from approximately 200 mg field-moist soil by using the D6005 Fungal/Bacterial Miniprep set (Zymo Research®) following the manufacturer’s instructions. To determine bacterial diversity and community composition, we employed a culture – independent approach of high-throughput pyrosequencing of the 16S rRNA gene. Metagenomic sequencing was made with Illumina MiSeq platform in Base Clear Company. The microbial component of soil plays a crucial role in cycling of nutrients in biosphere. Our study was a preliminary attempt at observing bacterial diversity in soil under two common but contrasting tillage practices. The number of sequenced reads obtained for PT (161 917) was higher than DP (131 194). The 10 most abundant genus in soil sample were the same (Arthrobacter, Candidatus Saccharibacteria, Actinobacteria, Acidobacterium, Mycobacterium, Bacillus, Alphaproteobacteria, Longilinea, Gemmatimonas, Solirubrobacter), just the percent of community part was different. In DP the Arthrobacter and Acidobacterium consist respectively 8.4 % and 2.5%, meanwhile in PT just 5.8% and 2.1% of all community. The Nocardioides and Terrabacter were observed just in PT. This work was supported by the project VP1-3.1-ŠMM-01-V-03-001 NKPDOKT and National Science Program: The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems [grant number SIT-9/2015] funded by the Research Council of Lithuania.

Keywords: deep ploughing, metagenomics, ploughless tillage, soil community analysis

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120 E-Business Role in the Development of the Economy of Sultanate of Oman

Authors: Mairaj Salim, Asma Zaheer

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Oman has accomplished as much or more than its fellow Gulf monarchies, despite starting from scratch considerably later, having less oil income to utilize, dealing with a larger and more rugged geography, and resolving a bitter civil war along the way. Of course, Oman's progress in the past 30-plus years has not been without problems and missteps, but the balance is squarely on the positive side of the ledger. Oil has been the driving force of the Omani economy since Oman began commercial production in 1967. The oil industry supports the country’s high standard of living and is primarily responsible for its modern and expansive infrastructure, including electrical utilities, telephone services, roads, public education and medical services. In addition to extensive oil reserves, Oman also has substantial natural gas reserves, which are expected to play a leading role in the Omani economy in the Twenty-first Century. To reduce the country’s dependence on oil revenues, the government is restructuring the economy by directing investment to non-oil activities. Since the 21st century IT has changed the performing tasks. To manage the affairs for the benefits of organizations and economy, the Omani government has adopted E-Business technologies for the development. E-Business is important because it allows • Transformation of old economy relationships (vertical/linear relationships) to new economy relationships characterized by end-to-end relationship management solutions (integrated or extended relationships) • Facilitation and organization of networks, small firms depend on ‘partner’ firms for supplies and product distribution to meet customer demands • SMEs to outsource back-end process or cost centers enabling the SME to focus on their core competence • ICT to connect, manage and integrate processes internally and externally • SMEs to join networks and enter new markets, through shortened supply chains to increase market share, customers and suppliers • SMEs to take up the benefits of e-business to reduce costs, increase customer satisfaction, improve client referral and attract quality partners • New business models of collaboration for SMEs to increase their skill base • SMEs to enter virtual trading arena and increase their market reach A national strategy for the advancement of information and communication technology (ICT) has been worked out, mainly to introduce e-government, e-commerce, and a digital society. An information technology complex KOM (Knowledge Oasis Muscat) had been established, consisting of section for information technology, incubator services, a shopping center of technology software and hardware, ICT colleges, E-Government services and other relevant services. So, all these efforts play a vital role in the development of Oman economy.

Keywords: ICT, ITA, CRM, SCM, ERP, KOM, SMEs, e-commerce and e-business

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119 Purple Spots on Historical Parchments: Confirming the Microbial Succession at the Basis of Biodeterioration

Authors: N. Perini, M. C. Thaller, F. Mercuri, S. Orlanducci, A. Rubechini, L. Migliore

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The preservation of cultural heritage is one of the major challenges of today’s society, because of the fundamental right of future generations to inherit it as the continuity with their historical and cultural identity. Parchments, consisting of a semi-solid matrix of collagen produced from animal skin (i.e., sheep or goats), are a significant part of the cultural heritage, being used as writing material for many centuries. Due to their animal origin, parchments easily undergo biodeterioration. The most common biological damage is characterized by isolated or coalescent purple spots that often leads to the detachment of the superficial layer and the loss of the written historical content of the document. Although many parchments with the same biodegradative features were analyzed, no common causative agent has been found so far. Very recently, a study was performed on a purple-damaged parchment roll dated back 1244 A.D, the A.A. Arm. I-XVIII 3328, belonging to the oldest collection of the Vatican Secret Archive (Fondo 'Archivum Arcis'), by comparing uncolored undamaged and purple damaged areas of the same document. As a whole, the study gave interesting results to hypothesize a model of biodeterioration, consisting of a microbial succession acting in two main phases: the first one, common to all the damaged parchments, is characterized by halophilic and halotolerant bacteria fostered by the salty environment within the parchment maybe induced by bringing of the hides; the second one, changing with the individual history of each parchment, determines the identity of its colonizers. The design of this model was pivotal to this study, performed by different labs of the Tor Vergata University (Rome, Italy), in collaboration with the Vatican Secret Archive. Three documents, belonging to a collection of dramatically damaged parchments archived as 'Faldone Patrizi A 19' (dated back XVII century A.D.), were analyzed through a multidisciplinary approach, including three updated technologies: (i) Next Generation Sequencing (NGS, Illumina) to describe the microbial communities colonizing the damaged and undamaged areas, (ii) RAMAN spectroscopy to analyze the purple pigments, (iii) Light Transmitted Analysis (LTA) to evaluate the kind and entity of the damage to native collagen. The metagenomic analysis obtained from NGS revealed DNA sequences belonging to Halobacterium salinarum mainly in the undamaged areas. RAMAN spectroscopy detected pigments within the purple spots, mainly bacteriorhodopsine/rhodopsin-like pigments, a purple transmembrane protein containing retinal and present in Halobacteria. The LTA technique revealed extremely damaged collagen structures in both damaged and undamaged areas of the parchments. In the light of these data, the study represents a first confirmation of the microbial succession model described above. The demonstration of this model is pivotal to start any possible new restoration strategy to bring back historical parchments to their original beauty, but also to open opportunities for intervention on a huge amount of documents.

Keywords: biodeterioration, parchments, purple spots, ecological succession

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118 Entrepreneurial Dynamism and Socio-Cultural Context

Authors: Shailaja Thakur

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Managerial literature abounds with discussions on business strategies, success stories as well as cases of failure, which provide an indication of the parameters that should be considered in gauging the dynamism of an entrepreneur. Neoclassical economics has reduced entrepreneurship to a mere factor of production, driven solely by the profit motive, thus stripping him of all creativity and restricting his decision making to mechanical calculations. His ‘dynamism’ is gauged simply by the amount of profits he earns, marginalizing any discussion on the means that he employs to attain this objective. With theoretical backing, we have developed an Index of Entrepreneurial Dynamism (IED) giving weights to the different moves that the entrepreneur makes during his business journey. Strategies such as changes in product lines, markets and technology are gauged as very important (weighting of 4); while adaptations in terms of technology, raw materials used, upgradations in skill set are given a slightly lesser weight of 3. Use of formal market analysis, diversification in related products are considered moderately important (weight of 2) and being a first generation entrepreneur, employing managers and having plans to diversify are taken to be only slightly important business strategies (weight of 1). The maximum that an entrepreneur can score on this index is 53. A semi-structured questionnaire is employed to solicit the responses from the entrepreneurs on the various strategies that have been employed by them during the course of their business. Binary as well as graded responses are obtained, weighted and summed up to give the IED. This index was tested on about 150 tribal entrepreneurs in Mizoram, a state of India and was found to be highly effective in gauging their dynamism. This index has universal acceptability but is devoid of the socio-cultural context, which is very central to the success and performance of the entrepreneurs. We hypothesize that a society that respects risk taking takes failures in its stride, glorifies entrepreneurial role models, promotes merit and achievement is one that has a conducive socio- cultural environment for entrepreneurship. For obtaining an idea about the social acceptability, we are putting forth questions related to the social acceptability of business to another set of respondents from different walks of life- bureaucracy, academia, and other professional fields. Similar weighting technique is employed, and index is generated. This index is used for discounting the IED of the respondent entrepreneurs from that region/ society. This methodology is being tested for a sample of entrepreneurs from two very different socio- cultural milieus- a tribal society and a ‘mainstream’ society- with the hypothesis that the entrepreneurs in the tribal milieu might be showing a higher level of dynamism than their counterparts in other regions. An entrepreneur who scores high on IED and belongs to society and culture that holds entrepreneurship in high esteem, might not be in reality as dynamic as a person who shows similar dynamism in a relatively discouraging or even an outright hostile environment.

Keywords: index of entrepreneurial dynamism, India, social acceptability, tribal entrepreneurs

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117 Interferon-Induced Transmembrane Protein-3 rs12252-CC Associated with the Progress of Hepatocellular Carcinoma by Up-Regulating the Expression of Interferon-Induced Transmembrane Protein 3

Authors: Yuli Hou, Jianping Sun, Mengdan Gao, Hui Liu, Ling Qin, Ang Li, Dongfu Li, Yonghong Zhang, Yan Zhao

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Background and Aims: Interferon-induced transmembrane protein 3 (IFITM3) is a component of ISG (Interferon-Stimulated Gene) family. IFITM3 has been recognized as a key signal molecule regulating cell growth in some tumors. However, the function of IFITM3 rs12252-CC genotype in the hepatocellular carcinoma (HCC) remains unknown to author’s best knowledge. A cohort study was employed to clarify the relationship between IFITM3 rs12252-CC genotype and HCC progression, and cellular experiments were used to investigate the correlation of function of IFITM3 and the progress of HCC. Methods: 336 candidates were enrolled in study, including 156 with HBV related HCC and 180 with chronic Hepatitis B infections or liver cirrhosis. Polymerase chain reaction (PCR) was employed to determine the gene polymorphism of IFITM3. The functions of IFITM3 were detected in PLC/PRF/5 cell with different treated:LV-IFITM3 transfected with lentivirus to knockdown the expression of IFITM3 and LV-NC transfected with empty lentivirus as negative control. The IFITM3 expression, proliferation and migration were detected by Quantitative reverse transcription polymerase chain reaction (qRT-PCR), QuantiGene Plex 2.0 assay, western blotting, immunohistochemistry, Cell Counting Kit(CCK)-8 and wound healing respectively. Six samples (three infected with empty lentiviral as control; three infected with LV-IFITM3 vector lentiviral as experimental group ) of PLC/PRF/5 were sequenced at BGI (Beijing Genomics Institute, Shenzhen,China) using RNA-seq technology to identify the IFITM3-related signaling pathways and chose PI3K/AKT pathway as related signaling to verify. Results: The patients with HCC had a significantly higher proportion of IFITM3 rs12252-CC compared with the patients with chronic HBV infection or liver cirrhosis. The distribution of CC genotype in HCC patients with low differentiation was significantly higher than that in those with high differentiation. Patients with CC genotype found with bigger tumor size, higher percentage of vascular thrombosis, higher distribution of low differentiation and higher 5-year relapse rate than those with CT/TT genotypes. The expression of IFITM3 was higher in HCC tissues than adjacent normal tissues, and the level of IFITM3 was higher in HCC tissues with low differentiation and metastatic than high/medium differentiation and without metastatic. Higher RNA level of IFITM3 was found in CC genotype than TT genotype. In PLC/PRF/5 cell with knockdown, the ability of cell proliferation and migration was inhibited. Analysis RNA sequencing and verification of RT-PCR found out the phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin(PI3K/AKT/mTOR) pathway was associated with knockdown IFITM3.With the inhibition of IFITM3, the expression of PI3K/AKT/mTOR signaling pathway was blocked and the expression of vimentin was decreased. Conclusions: IFITM3 rs12252-CC with the higher expression plays a vital role in the progress of HCC by regulating HCC cell proliferation and migration. These effects are associated with PI3K/AKT/mTOR signaling pathway.

Keywords: IFITM3, interferon-induced transmembrane protein 3, HCC, hepatocellular carcinoma, PI3K/ AKT/mTOR, phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin

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116 Isolation and Identification of Sarcocystis suihominis in a Slaughtered Domestic Pig (Sus scrofa) in Benue State, Nigeria

Authors: H. I. Obadiah, S. N. Wieser, E. A. Omudu, B. O. Atu, O. Byanet, L. Schnittger, M. Florin-Christensen

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Sarcocystis sp. are Apicomplexan protozoan parasites with a life cycle that involves a predator and a prey as final and intermediate hosts, respectively. In tissues of the intermediate hosts, the parasites produce sarcocysts that vary in size and morphology according to the species. When a suitable predator ingests sarcocyst-containing meat, the parasites are released in the intestine and undergo sexual reproduction producing infective sporocysts, which are excreted with the feces into the environment. The cycle is closed when a prey ingests sporocyst-contaminated water or pasture; the parasites gain access to the circulation, and eventually invade tissues and reproduce asexually yielding sarcocysts. Pig farming is a common practice in Nigeria as well as in many countries around the world. In addition to its importance as protein source, pork is also a source of several pathogens relevant to humans. In the case of Sarcocystis, three species have been described both in domestic and wild pigs, namely, S. miescheriana, S. porcifelis and S. suihominis. Humans can act both as final and aberrant intermediate hosts of S. suihominis, after ingesting undercooked sarcocyst-infested pork. Infections are usually asymptomatic but can be associated with inappetence, nausea, vomiting and diarrhea, or with muscle pain, fever, eosinophilia and bronchospasm, in humans acting as final or intermediate hosts, respectively. Moreover, excretion of infective forms with human feces leads to further dissemination of the infection. In this study, macroscopic sarcocysts of white color, oval shape and a size range of approximately 3-5 mm were observed in the skeletal muscle of a slaughtered pig in an abattoir in Makurdi, Benue State, Nigeria, destined to human consumption. Sarcocysts were excised and washed in distilled water, and genomic DNA was extracted using a commercial kit. The near-complete length of the 18S rRNA gene was analyzed after PCR amplification of two overlapping fragments, each of which were submitted to direct sequencing. In addition, the mitochondrial cytochrome oxidase (cox-1) gene was PCR-amplified and directly sequenced. Two phylogenetic trees containing the obtained sequences along with available relevant 18S rRNA and cox-1 sequences were constructed by neighbor joining after alignment, using the corresponding sequences of Toxoplasma gondii as outgroup. The results showed in both cases that the analyzed sequences grouped with S. suihominis with high bootstrap value, confirming the identity of this macroscopic sarcocyst-forming parasite as S. suihominis. To the best of our knowledge, these results represent the first demonstration of this parasite in pigs of Nigeria and the largest sarcocysts described so far for S. suihominis. The close proximity between pigs and humans in pig farms, and the frequent poor sanitary conditions in human dwellings strongly suggest that the parasite undergoes the sexual stages of its life cycle in humans as final hosts. These findings provide an important reference for the examination and control of Sarcocystis species in pigs of Nigeria.

Keywords: nigeria, pork, sarcocystis suihominis, zoonotic parasite

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115 Effects of Glucogenic and Lipogenic Diets on Ruminal Microbiota and Metabolites in Vitro

Authors: Beihai Xiong, Dengke Hua, Wouter Hendriks, Wilbert Pellikaan

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To improve the energy status of dairy cows in the early lactation, lots of jobs have been done on adjusting the starch to fiber ratio in the diet. As a complex ecosystem, the rumen contains a large population of microorganisms which plays a crucial role in feed degradation. Further study on the microbiota alterations and metabolic changes under different dietary energy sources is essential and valuable to better understand the function of the ruminal microorganisms and thereby to optimize the rumen function and enlarge feed efficiency. The present study will focus on the effects of two glucogenic diets (G: ground corn and corn silage; S: steam-flaked corn and corn silage) and a lipogenic diet (L: sugar beet pulp and alfalfa silage) on rumen fermentation, gas production, the ruminal microbiota and metabolome, and also their correlations in vitro. The gas production was recorded consistently, and the gas volume and producing rate at times 6, 12, 24, 48 h were calculated separately. The fermentation end-products were measured after fermenting for 48 h. The ruminal bacteria and archaea communities were determined by 16S RNA sequencing technique, the metabolome profile was tested through LC-MS methods. Compared to the diet G and S, the L diet had a lower dry matter digestibility, propionate production, and ammonia-nitrogen concentration. The two glucogenic diets performed worse in controlling methane and lactic acid production compared to the L diet. The S diet produced the greatest cumulative gas volume at any time points during incubation compared to the G and L diet. The metabolic analysis revealed that the lipid digestion was up-regulated by the diet L than other diets. On the subclass level, most metabolites belonging to the fatty acids and conjugates were higher, but most metabolites belonging to the amino acid, peptides, and analogs were lower in diet L than others. Differences in rumen fermentation characteristics were associated with (or resulting from) changes in the relative abundance of bacterial and archaeal genera. Most highly abundant bacteria were stable or slightly influenced by diets, while several amylolytic and cellulolytic bacteria were sensitive to the dietary changes. The L diet had a significantly higher number of cellulolytic bacteria, including the genera of Ruminococcus, Butyrivibrio, Eubacterium, Lachnospira, unclassified Lachnospiraceae, and unclassified Ruminococcaceae. The relative abundances of amylolytic bacteria genera including Selenomonas_1, Ruminobacter, and Succinivibrionaceae_UCG-002 were higher in diet G and S. These affected bacteria was also proved to have high associations with certain metabolites. The Selenomonas_1 and Succinivibrionaceae_UCG-002 may contribute to the higher propionate production in the diet G and S through enhancing the succinate pathway. The results indicated that the two glucogenic diets had a greater extent of gas production, a higher dry matter digestibility, and produced more propionate than diet L. The steam-flaked corn did not show a better performance on fermentation end-products than ground corn. This study has offered a deeper understanding of ruminal microbial functions which could assistant the improvement in rumen functions and thereby in the ruminant production.

Keywords: gas production, metabolome, microbiota, rumen fermentation

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114 Clinical Cases of Rare Types of 'Maturity Onset Diabetes of the Young' Diabetes

Authors: Alla Ovsyannikova, Oksana Rymar, Elena Shakhtshneider, Mikhail Voevoda

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In Siberia endocrinologists increasingly noted young patients with the course of diabetes mellitus differing from 1 and 2 types. Therefore we did a molecular genetic study for this group of patients to verify the monogenic forms of diabetes mellitus in them and researched the characteristics of this pathology. When confirming the monogenic form of diabetes, we performed a correction therapy for many patients (transfer from insulin to tablets), prevented specific complications, examined relatives and diagnosed their diabetes at the preclinical stage, revealed phenotypic characteristics of the pathology which led to the high significance of this work. Materials and Methods: We observed 5 patients (4 families). We diagnosed MODY (Maturity Onset Diabetes of the Young) during the molecular genetic testing (direct automatic sequencing). All patients had a full clinical examination, blood samples for biochemical research, determination of C-peptide and TSH, antibodies to b-cells, microalbuminuria, abdominal ultrasound, heart and thyroid ultrasound, examination of ophthalmologist. Results: We diagnosed 3 rare types of MODY: two women had MODY8, one man – MODY6 and man and his mother - MODY12. Patients with types 8 and 12 had clinical features. Age of onset hyperglycemia ranged from 26 to 34 years. In a patient with MODY6 fasting hyperglycemia was detected during a routine examination. Clinical symptoms, complications were not diagnosed. The patient observes a diet. In the first patient MODY8 was detected during first pregnancy, she had itchy skin and mostly postprandial hyperglycemia. Upon examination we determined glycated hemoglobin 7.5%, retinopathy, non-proliferative stage, peripheral neuropathy. She uses a basic bolus insulin therapy. The second patient with MODY8 also had clinical manifestations of hyperglycemia (pruritus, thirst), postprandial hyperglycemia and diabetic nephropathy, a stage of microalbuminuria. The patient was diagnosed autoimmune thyroiditis. She used inhibitors of DPP-4. The patient with MODY12 had an aggressive course. In the detection of hyperglycemia he had complaints of visual impairment, intense headaches, leg cramps. The patient had a history of childhood convulsive seizures of non-epileptic genesis, without organic pathology, which themselves were stopped at the age of 12 years. When we diagnosed diabetes a patient was 28 years, he had hypertriglyceridemia, atherosclerotic plaque in the carotid artery, proliferative retinopathy (lacerocoagulation). Diabetes and early myocardial infarction were observed in three cases in family. We prescribe therapy with sulfonylureas and SGLT-2 inhibitors with a positive effect. At the patient's mother diabetes began at a later age (30 years) and a less aggressive course was observed. She also has hypertriglyceridemia and uses oral hypoglycemic drugs. Conclusions: 1) When young patients with hyperglycemia have extrapancreatic pathologies and diabetic complications with a short duration of diabetes we can assume they have one of type of MODY diabetes. 2) In patients with monogenic forms of diabetes mellitus, the clinical manifestations of hyperglycemia in each succeeding generation are revealed at an earlier age. Research had increased our knowledge of the monogenic forms of diabetes. The reported study was supported by RSCF, research project No. 14-15-00496-P.

Keywords: diabetes mellitus, MODY diabetes, monogenic forms, young patients

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113 A Comparative Study on Compliment Response between Indonesian EFL Students and English Native Speakers

Authors: Maria F. Seran

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In second language interaction, an EFL student always carries his knowledge of targeted language and sometimes gets influenced by his first language cultures which makes him transfer his utterances from the first language to the second language. The influence of L1 cultures somehow can lead to face-threatening act when it comes to responding on speech act, for instance, compliment. A speaker praises a compliment to show gratitude, and in return, he expects for compliment respond uttered by the hearer. While Western people use more acceptance continuum on compliment response, Indonesians utter more denial continuum which can somehow put the speakers into a face-threating situation and offense. This study investigated compliment response employed by EFL students and English native speakers. The study was distinct as none compliment response studies had been conducted to compare the compliment response between English native speakers and two different Indonesian EFL proficiency groups in which this research sought to meet this need. This study was significant for EFL teachers because it gave insight on cross-cultural understanding and brought pedagogical implication on explicit pragmatic instruction. Two research questions were set, 1. How do Indonesian EFL students and English native speakers respond compliments? 2. Is there any correlation between Indonesia EFL students’ proficiency and their compliment response use in English? The study involved three groups of participants; 5 English native speakers, 10 high-proficiency and 10 low-proficiency Indonesian EFL university students. The research instruments used in this study were as follows, an online TOEFL prediction test, focusing on grammar skill which was modified from Barron TOEFL exercise test, and a discourse completion task (DCT), consisting of 10 compliment respond items. Based on the research invitation, 20 second-year university students majoring in English education at Widya Mandira Catholic University, Kupang, East Nusa Tenggara, Indonesia who willingly participated in the research took the TOEFL prediction test online from the link provided. Students who achieved score 75-100 in test were categorized as high-proficiency students, while, students who attained score below 74 were considered as low-proficiency students. Then, the DCT survey was administered to these EFL groups and the native speaker group. Participants’ responses were coded and analyzed using categories of compliment response framework proposed by Tran. The study found out that 5 native speakers applied more compliment upgrades and appreciation token in compliment response, whereas, Indonesian EFL students combined some compliment response strategies in their utterance, such as, appreciation token, return and compliment downgrade. There is no correlation between students’ proficiency level and their CR responds as most EFL students in both groups produced less varied compliment responses and only 4 Indonesian high-proficiency students uttered more varied and were similar to the native speakers. The combination strategies used by EFL students can be explained as the influence of pragmatic transfer from L1 to L2; therefore, EFL teachers should explicitly teach more compliment response strategies to raise students’ awareness on English culture and elaborate their speaking to be more competence as close to native speakers as possible.

Keywords: compliment response, English native speakers, Indonesian EFL students, speech acts

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112 Improving Literacy Level Through Digital Books for Deaf and Hard of Hearing Students

Authors: Majed A. Alsalem

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In our contemporary world, literacy is an essential skill that enables students to increase their efficiency in managing the many assignments they receive that require understanding and knowledge of the world around them. In addition, literacy enhances student participation in society improving their ability to learn about the world and interact with others and facilitating the exchange of ideas and sharing of knowledge. Therefore, literacy needs to be studied and understood in its full range of contexts. It should be seen as social and cultural practices with historical, political, and economic implications. This study aims to rebuild and reorganize the instructional designs that have been used for deaf and hard-of-hearing (DHH) students to improve their literacy level. The most critical part of this process is the teachers; therefore, teachers will be the center focus of this study. Teachers’ main job is to increase students’ performance by fostering strategies through collaborative teamwork, higher-order thinking, and effective use of new information technologies. Teachers, as primary leaders in the learning process, should be aware of new strategies, approaches, methods, and frameworks of teaching in order to apply them to their instruction. Literacy from a wider view means acquisition of adequate and relevant reading skills that enable progression in one’s career and lifestyle while keeping up with current and emerging innovations and trends. Moreover, the nature of literacy is changing rapidly. The notion of new literacy changed the traditional meaning of literacy, which is the ability to read and write. New literacy refers to the ability to effectively and critically navigate, evaluate, and create information using a range of digital technologies. The term new literacy has received a lot of attention in the education field over the last few years. New literacy provides multiple ways of engagement, especially to those with disabilities and other diverse learning needs. For example, using a number of online tools in the classroom provides students with disabilities new ways to engage with the content, take in information, and express their understanding of this content. This study will provide teachers with the highest quality of training sessions to meet the needs of DHH students so as to increase their literacy levels. This study will build a platform between regular instructional designs and digital materials that students can interact with. The intervention that will be applied in this study will be to train teachers of DHH to base their instructional designs on the notion of Technology Acceptance Model (TAM) theory. Based on the power analysis that has been done for this study, 98 teachers are needed to be included in this study. This study will choose teachers randomly to increase internal and external validity and to provide a representative sample from the population that this study aims to measure and provide the base for future and further studies. This study is still in process and the initial results are promising by showing how students have engaged with digital books.

Keywords: deaf and hard of hearing, digital books, literacy, technology

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111 The First Complete Mitochondrial Genome of Melon Thrips, Thrips palmi (Thripinae: Thysanoptera): Vector for Tospoviruses

Authors: Kaomud Tyagi, Rajasree Chakraborty, Shantanu Kundu, Devkant Singha, Kailash Chandra, Vikas Kumar

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The melon thrips, Thrips palmi is a serious pest of a wide range of agriculture crops and also act as vectors for plant viruses (genus Tospovirus, family Bunyaviridae). More molecular data on this species is required to understand the cryptic speciation and evolutionary affiliations. Mitochondrial genomes have been widely used in phylogenetic and evolutionary studies in insect. So far, mitogenomes of five thrips species (Anaphothrips obscurus, Frankliniella intonsa, Frankliniella occidentalis, Scirtothrips dorsalis and Thrips imaginis) is available in the GenBank database. In this study, we sequenced the first complete mitogenome T. palmi and compared it with available thrips mitogenomes. We assembled the mitogenome from the whole genome sequencing data generated using Illumina Hiseq2500. Annotation was performed using MITOS web-server to estimate the location of protein coding genes (PCGs), transfer RNA (tRNAs), ribosomal RNAs (rRNAs) and their secondary structures. The boundaries of PCGs and rRNAs was confirmed manually in NCBI. Phylogenetic analyses were performed using the 13 PCGs data using maximum likelihood (ML) in PAUP, and Bayesian inference (BI) in MrBayes 3.2. The complete mitogenome of T. palmi was 15,333 base pairs (bp), which was greater than the genomes of A. obscurus (14,890bp), F. intonsa (15,215 bp), F. occidentalis (14,889 bp) and S. dorsalis South Asia strain (SA1) (14,283 bp), but smaller than the genomes of T. imaginis (15,407 bp) and S. dorsalis East Asia strain (EA1) (15,343bp). Like in other thrips species, the mitochondrial genome of T. palmi was represented by 37 genes, including 13 PCGs, large and small ribosomal RNA (rrnL and rrnS) genes, 22 transfer RNA (tRNAs) genes (with one extra gene for trn-Serine) and two A+T-rich control regions (CR1 and CR2). Thirty one genes were observed on heavy (H) strand and six genes on the light (L) strand. The six tRNA genes (trnG,trnK, trnY, trnW, trnF, and trnH) were found to be conserved in all thrips species mitogenomes in their locations relative to a protein-coding or rRNA gene upstream or downstream. The gene arrangements of T. palmi is very close to T. imaginis except the rearrangements in tRNAs genes: trnR (arginine), and trnE (glutamic acid) were found to be located between cox3 and CR2 in T. imaginis which were translocated between atp6 and CR1 in T. palmi; trnL1 (Leucine) and trnS1(Serine) were located between atp6 and CR1 in T. imaginis which were translocated between cox3 and CR2 in T. palmi. The location of CR1 upstream of nad5 gene was suggested to be ancestral condition of the thrips species in subfamily Thripinae, was also observed in T. palmi. Both the Maximum likelihood (ML) and Bayesian Inference (BI) phylogenetic trees generated resulted in similar topologies. The T. palmi was clustered with T. imaginis. We concluded that more molecular data on the diverse thrips species from different hierarchical level is needed, to understand the phylogenetic and evolutionary relationships among them.

Keywords: thrips, comparative mitogenomics, gene rearrangements, phylogenetic analysis

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110 Using the Clinical Decision Support Platform, Dem DX, to Assess the ‘Urgent Community Care Team’s Notes Regarding Clinical Assessment, Management, and Healthcare Outcomes

Authors: R. Tariq, R. Lee

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Background: Heywood, Middleton & Rochdale Urgent Community Care Team (UCCT)1 is a great example of using a multidisciplinary team to cope with demand. The service reduces unnecessary admissions to hospitals and ensures that patients can leave the hospital quicker by making care more readily available within the community and patient’s homes. The team comprises nurses, community practitioners, and allied health professions, including physiotherapy, occupational therapy, pharmacy, and GPs. The main challenge for a team with a range of experiences and skill sets is to maintain consistency of care, which technology can help address. Allied healthcare professionals (HCPs) are often used in expanded roles with duties mainly involving patient consultations and decision making to ease pressure on doctors. The Clinical Reasoning Platform (CRP) Dem Dx is used to support new as well as experienced professionals in the decision making process. By guiding HCPs through diagnosing patients from an expansive directory of differential diagnoses, patients can receive quality care in the community. Actions on the platform are determined using NICE guidelines along with local guidance influencing the assessment and management of a patient. Objective: To compare the clinical assessment, decisions, and actions taken by the UCCT multidisciplinary team in the community and Dem Dx, using retrospective clinical cases. Methodology: Dem Dx was used to analyse 192 anonymised cases provided by the HMR UCCT. The team’s performance was compared with Dem Dx regarding the quality of the documentation of the clinical assessment and the next steps on the patient’s journey, including the initial management, actions, and any onward referrals made. The cases were audited by two medical doctors. Results: The study found that the actions outlined by the Dem Dx platform were appropriate in almost 87% of cases. When in a direct comparison between DemDX and the actions taken by the clinical team, it was found that the platform was suitable 83% (p<0.001) of the time and could lead to a potential improvement of 66% in the assessment and management of cases. Dem Dx also served to highlight the importance of comprehensive and high quality clinical documentation. The quality of documentation of cases by UCCT can be improved to provide a detailed account of the assessment and management process. By providing step-by-step guidance and documentation at every stage, Dem Dx may ensure that legal accountability has been fulfilled. Conclusion: With the ever expanding workforce in the NHS, technology has become a key component in driving healthcare outcomes. To improve healthcare provision and clinical reasoning, a decision support platform can be integrated into HCPs’ clinical practice. Potential assistance with clinical assessments, the most appropriate next step and actions in a patient’s care, and improvements in the documentation was highlighted by this retrospective study. A further study has been planned to ascertain the effectiveness of improving outcomes using the clinical reasoning platform within the clinical setting by clinicians.

Keywords: allied health professional, assessment, clinical reasoning, clinical records, clinical decision-making, ocumentation

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109 In Vitro Studies on Antimicrobial Activities of Lactic Acid Bacteria Isolated from Fresh Fruits for Biocontrol of Pathogens

Authors: Okolie Pius Ifeanyi, Emerenini Emilymary Chima

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Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits using Molecular Nested PCR analysis and the efficacy of cell free supernatants from Lactic Acid Bacteria (LAB) isolated from fresh fruits for in vitro control of some tomato pathogens. Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples. The inhibitory potentials of supernatant obtained from LAB isolates of fruits origin that were molecularly characterized were investigated against some tomato phytopathogens using agar-well method with the view to develop biological agents for some tomato disease causing organisms. Methodology: Gram positive, catalase negative strains of LAB were isolated from fresh fruits on Man Rogosa and Sharpe agar (Lab M) using streaking method. Isolates obtained were molecularly characterized by means of genomic DNA extraction kit (Norgen Biotek, Canada) method. Standard methods were used for Nested Polymerase Chain Reaction (PCR) amplification targeting the 16S rRNA gene using universal 16S rRNA gene and LAB specific primers, agarose gel electrophoresis, purification and sequencing of generated Nested PCR products (Macrogen Inc., USA). The partial sequences obtained were identified by blasting in the non-redundant nucleotide database of National Center for Biotechnology Information (NCBI). The antimicrobial activities of characterized LAB against some tomato phytopathogenic bacteria which include (Xanthomonas campestries, Erwinia caratovora, and Pseudomonas syringae) were obtained by using the agar well diffusion method. Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%). The cell free supernatants of LAB from fresh fruits origin (Weissella cibaria, Weissella confusa, Leuconostoc paramensenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Lactobacillus pentosus) can inhibits these bacteria by creating clear zones of inhibition around the wells containing cell free supernatants of the above mentioned strains of lactic acid bacteria. Conclusion: This study shows that potentially LAB can be quickly characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer. Tomato disease causing organisms can be most likely biologically controlled by using extracts from LAB. This finding will reduce the potential hazard from the use of chemical herbicides on plant.

Keywords: nested pcr, molecular characterization, 16s rRNA gene, lactic acid bacteria

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108 Interpersonal Competence Related to the Practice Learning of Occupational Therapy Students in Hong Kong

Authors: Lik Hang Gary Wong

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Background: Practice learning is crucial for preparing the healthcare profession to meet the real challenge upon graduation. Students are required to demonstrate their competence in managing interpersonal challenges, such as teamwork with other professionals and communicating well with the service users, during the placement. Such competence precedes clinical practice, and it may eventually affect students' actual performance in a clinical context. Unfortunately, there were limited studies investigating how such competence affects students' performance in practice learning. Objectives: The aim of this study is to investigate how self-rated interpersonal competence affects students' actual performance during clinical placement. Methods: 40 occupational therapy students from Hong Kong were recruited in this study. Prior to the clinical placement (level two or above), they completed an online survey that included the Interpersonal Communication Competence Scale (ICCS) measuring self-perceived competence in interpersonal communication. Near the end of their placement, the clinical educator rated students’ performance with the Student Practice Evaluation Form - Revised edition (SPEF-R). The SPEF-R measures the eight core competency domains required for an entry-level occupational therapist. This study adopted the cross-sectional observational design. Pearson correlation and multiple regression are conducted to examine the relationship between students' interpersonal communication competence and their actual performance in clinical placement. Results: The ICCS total scores were significantly correlated with all the SPEF-R domains, with correlation coefficient r ranging from 0.39 to 0.51. The strongest association was found with the co-worker communication domain (r = 0.51, p < 0.01), followed by the information gathering domain (r = 0.50, p < 0.01). Regarding the ICCS total scores as the independent variable and the rating in various SPEF-R domains as the dependent variables in the multiple regression analyses, the interpersonal competence measures were identified as a significant predictor of the co-worker communication (R² = 0.33, β = 0.014, SE = 0.006, p = 0.026), information gathering (R² = 0.27, β = 0.018, SE = 0.007, p = 0.011), and service provision (R² = 0.17, β = 0.017, SE = 0.007, p = 0.020). Moreover, some specific communication skills appeared to be especially important to clinical practice. For example, immediacy, which means whether the students were readily approachable on all social occasions, correlated with all the SPEF-R domains, with r-values ranging from 0.45 to 0.33. Other sub-skills, such as empathy, interaction management, and supportiveness, were also found to be significantly correlated to most of the SPEF-R domains. Meanwhile, the ICCS scores correlated differently with the co-worker communication domain (r = 0.51, p < 0.01) and the communication with the service user domain (r = 0.39, p < 0.05). It suggested that different communication skill sets would be required for different interpersonal contexts within the workplace. Conclusion: Students' self-perceived interpersonal communication competence could predict their actual performance during clinical placement. Moreover, some specific communication skills were more important to the co-worker communication but not to the daily interaction with the service users. There were implications on how to better prepare the students to meet the future challenge upon graduation.

Keywords: interpersonal competence, clinical education, healthcare professional education, occupational therapy, occupational therapy students

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107 Research Project on Learning Rationality in Strategic Behaviors: Interdisciplinary Educational Activities in Italian High Schools

Authors: Giovanna Bimonte, Luigi Senatore, Francesco Saverio Tortoriello, Ilaria Veronesi

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The education process considers capabilities not only to be seen as a means to a certain end but rather as an effective purpose. Sen's capability approach challenges human capital theory, which sees education as an ordinary investment undertaken by individuals. A complex reality requires complex thinking capable of interpreting the dynamics of society's changes to be able to make decisions that can be rational for private, ethical and social contexts. Education is not something removed from the cultural and social context; it exists and is structured within it. In Italy, the "Mathematical High School Project" is a didactic research project is based on additional laboratory courses in extracurricular hours where mathematics intends to bring itself in a dialectical relationship with other disciplines as a cultural bridge between the two cultures, the humanistic and the scientific ones, with interdisciplinary educational modules on themes of strong impact in younger life. This interdisciplinary mathematics presents topics related to the most advanced technologies and contemporary socio-economic frameworks to demonstrate how mathematics is not only a key to reading but also a key to resolving complex problems. The recent developments in mathematics provide the potential for profound and highly beneficial changes in mathematics education at all levels, such as in socio-economic decisions. The research project is built to investigate whether repeated interactions can successfully promote cooperation among students as rational choice and if the skill, the context and the school background can influence the strategies choice and the rationality. A Laboratory on Game Theory as mathematical theory was conducted in the 4th year of the Mathematical High Schools and in an ordinary scientific high school of the Scientific degree program. Students played two simultaneous games of repeated Prisoner's Dilemma with an indefinite horizon, with two different competitors in each round; even though the competitors in each round will remain the same for the duration of the game. The results highlight that most of the students in the two classes used the two games with an immunization strategy against the risk of losing: in one of the games, they started by playing Cooperate, and in the other by the strategy of Compete. In the literature, theoretical models and experiments show that in the case of repeated interactions with the same adversary, the optimal cooperation strategy can be achieved by tit-for-tat mechanisms. In higher education, individual capacities cannot be examined independently, as conceptual framework presupposes a social construction of individuals interacting and competing, making individual and collective choices. The paper will outline all the results of the experimentation and the future development of the research.

Keywords: game theory, interdisciplinarity, mathematics education, mathematical high school

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106 Identification of Odorant Receptors through the Antennal Transcriptome of the Grapevine Pest, Lobesia botrana (Lepidoptera: Tortricidae)

Authors: Ricardo Godoy, Herbert Venthur, Hector Jimenez, Andres Quiroz, Ana Mutis

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In agriculture, grape production has great economic importance at global level, considering that in 2013 it reached 7.4 million hectares (ha) covered by plantations of this fruit worldwide. Chile is the number one exporter in the world with 800,000 tons. However, these values have been threatened by the attack of the grapevine moth, Lobesia botrana (Denis & Schiffermuller) (Lepidoptera: Tortricidae), since its detection in 2008. Nowadays, the use of semiochemicals, in particular the major component of the sex pheromone, (E,Z)-7.9-dodecadienil acetate, are part of mating disruption methods to control L. botrana. How insect pests can recognize these molecules, is being part of huge efforts to deorphanize their olfactory mechanism at molecular level. Thus, an interesting group of proteins has been identified in the antennae of insects, where odorant-binding proteins (OBPs) are known by transporting molecules to odorant receptors (ORs) and a co-receptor (ORCO) causing a behavioral change in the insect. Other proteins such as chemosensory proteins (CSPs), ionotropic receptors (IRs), odorant degrading enzymes (ODEs) and sensory neuron membrane proteins (SNMPs) seem to be involved, but few studies have been performed so far. The above has led to an increasing interest in insect communication at a molecular level, which has contributed to both a better understanding of the olfaction process and the design of new pest management strategies. To date, it has been reported that the ORs can detect one or a small group of odorants in a specific way. Therefore, the objective of this study is the identification of genes that encode these ORs using the antennal transcriptome of L. botrana. Total RNA was extracted for females and males of L. botrana, and the antennal transcriptome sequenced by Next Generation Sequencing service using an Illumina HiSeq2500 platform with 50 million reads per sample. Unigenes were assembled using Trinity v2.4.0 package and transcript abundance was obtained using edgeR. Genes were identified using BLASTN and BLASTX locally installed in a Unix system and based on our own Tortricidae database. Those Unigenes related to ORs were characterized using ORFfinder and protein Blastp server. Finally, a phylogenetic analysis was performed with the candidate amino acid sequences for LbotORs including amino acid sequences of other moths ORs, such as Bombyx mori, Cydia pomonella, among others. Our findings suggest 61 genes encoding ORs and one gene encoding an ORCO in both sexes, where the greatest difference was found in the OR6 because of the transcript abundance according to the value of FPKM in females and males was 1.48 versus 324.00. In addition, according to phylogenetic analysis OR6 is closely related to OR1 in Cydia pomonella and OR6, OR7 in Epiphyas postvittana, which have been described as pheromonal receptors (PRs). These results represent the first evidence of ORs present in the antennae of L. botrana and a suitable starting point for further functional studies with selected ORs, such as OR6, which is potentially related to pheromonal recognition.

Keywords: antennal transcriptome, lobesia botrana, odorant receptors (ORs), phylogenetic analysis

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105 Multisensory Science, Technology, Engineering and Mathematics Learning: Combined Hands-on and Virtual Science for Distance Learners of Food Chemistry

Authors: Paulomi Polly Burey, Mark Lynch

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It has been shown that laboratory activities can help cement understanding of theoretical concepts, but it is difficult to deliver such an activity to an online cohort and issues such as occupational health and safety in the students’ learning environment need to be considered. Chemistry, in particular, is one of the sciences where practical experience is beneficial for learning, however typical university experiments may not be suitable for the learning environment of a distance learner. Food provides an ideal medium for demonstrating chemical concepts, and along with a few simple physical and virtual tools provided by educators, analytical chemistry can be experienced by distance learners. Food chemistry experiments were designed to be carried out in a home-based environment that 1) Had sufficient scientific rigour and skill-building to reinforce theoretical concepts; 2) Were safe for use at home by university students and 3) Had the potential to enhance student learning by linking simple hands-on laboratory activities with high-level virtual science. Two main components of the resources were developed, a home laboratory experiment component, and a virtual laboratory component. For the home laboratory component, students were provided with laboratory kits, as well as a list of supplementary inexpensive chemical items that they could purchase from hardware stores and supermarkets. The experiments used were typical proximate analyses of food, as well as experiments focused on techniques such as spectrophotometry and chromatography. Written instructions for each experiment coupled with video laboratory demonstrations were used to train students on appropriate laboratory technique. Data that students collected in their home laboratory environment was collated across the class through shared documents, so that the group could carry out statistical analysis and experience a full laboratory experience from their own home. For the virtual laboratory component, students were able to view a laboratory safety induction and advised on good characteristics of a home laboratory space prior to carrying out their experiments. Following on from this activity, students observed laboratory demonstrations of the experimental series they would carry out in their learning environment. Finally, students were embedded in a virtual laboratory environment to experience complex chemical analyses with equipment that would be too costly and sensitive to be housed in their learning environment. To investigate the impact of the intervention, students were surveyed before and after the laboratory series to evaluate engagement and satisfaction with the course. Students were also assessed on their understanding of theoretical chemical concepts before and after the laboratory series to determine the impact on their learning. At the end of the intervention, focus groups were run to determine which aspects helped and hindered learning. It was found that the physical experiments helped students to understand laboratory technique, as well as methodology interpretation, particularly if they had not been in such a laboratory environment before. The virtual learning environment aided learning as it could be utilized for longer than a typical physical laboratory class, thus allowing further time on understanding techniques.

Keywords: chemistry, food science, future pedagogy, STEM education

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104 Analysis of Engagement Methods in the College Classroom Post Pandemic

Authors: Marsha D. Loda

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College enrollment is declining and generation Z, today’s college students, are struggling. Before the pandemic, researchers characterized this generational cohort as unique. Gen Z has been called the most achievement-oriented generation, as they enjoy greater economic status, are more racially and ethnically diverse, and better educated than any other generation. However, they are also the most likely generation to suffer from depression and anxiety. Gen Z has grown up largely with usually well-intentioned but overprotective parents who inadvertently kept them from learning life skills, likely impacting their ability to cope with and to effectively manage challenges. The unprecedented challenges resulting from the pandemic up ended their world and left them emotionally reeling. One of the ramifications of this for higher education is how to reengage current Gen Z students in the classroom. This research presents qualitative findings from 24 single-spaced pages of verbatim comments from college students. Research questions concerned what helps them learn and what they abhor, as well as how to engage them with the university outside of the classroom to aid in retention. Students leave little doubt about what they want to experience in the classroom. In order of mention, students want discussion, to engage with questions, to hear how a topic relates to real life and the real world, to feel connections with the professor and fellow students, and to have an opportunity to give their opinions. They prefer a classroom that involves conversation, with interesting topics and active learning. “professor talks instead of lecturing” “professor builds a connection with the classroom” “I am engaged because it feels like a respectful conversation” Similarly, students are direct about what they dislike in a classroom. In order of frequency, students dislike teachers unenthusiastically reading word or word from notes or presentations, repeating the text without adding examples, or addressing how to apply the information. “All lecture. I can read the book myself” “Not taught how to apply the skill or lesson” “Lectures the entire time. Lesson goes in one ear and out the other.” Pertaining to engagement outside the classroom, Gen Z challenges higher education to step outside the box. They don’t want to just hear from professionals in their field, they want to meet and interact with them. Perhaps because of their dependence on technology and pandemic isolation, they seem to reach out for assistance in forming social bonds. “I believe fun and social events are the best way to connect with students and get them involved. Cookouts, raffles, socials, or networking events would all most likely appeal to many students”. “Events… even if they aren’t directly related to learning. Maybe like movie nights… doing meet ups at restaurants”. Qualitative research suggests strategy. This research is rife with strategic implications to improve learning, increase engagement and reduce drop-out rates among Generation Z higher education students. It also compliments existing research on student engagement. With college enrollment declining by some 1.3 million students over the last two years, this research is both timely and important.

Keywords: college enrollment, generation Z, higher education, pandemic, student engagement

Procedia PDF Downloads 101
103 Phytoplankton Structure and Invasive Cyanobacterial Species of Polish Temperate Lakes: Their Associations with Environmental Parameters and Findings About Their Toxic Properties

Authors: Tumer Orhun Aykut, Robin Michael Crucitti-Thoo, Agnieszka Rudak, Iwona Jasser

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Due to eutrophication connected to the growing human population, intensive agriculture, industrialization, and reinforcement of global warming, freshwater resources are changing negatively in every region of the World. This change also concerns the replacement of native species by invasive ones that can spread in many ways. Biological invasions are a developing problem to ecosystem continuity and their presence is mostly common in freshwater bodies. The occurrence and potential invasion of the species depends on associations between abiotic and biotic variables. Due to climate change, many species can extend their range from low to high latitudes and differ in their geographic ranges. In addition, the hydrological issues strongly influence the physicochemical parameters and biological processes, especially the growth rates of species and bloom formation of Cyanobacteria. Among tropical invasive species noted in temperate Europe, Raphidiopsis raciborskii, Chrysosporum bergii, and Sphaerospermopsis aphanizomenoides are considered a serious threat. R. raciborskii being the most important one as it is already known as a highly invasive species in almost all around the World, is a freshwater, planktonic, filamentous, potentially toxic, and nitrogen-fixing Cyanobacteria. This study aimed to investigate the presence of invasive cyanobacterial species in temperate lakes in Northeastern Poland, reveal the composition of phytoplankton communities, determine the effect of environmental variables, and identify the toxic properties of invasive Cyanobacteria and other phytoplankton groups. Our study was conducted in twenty-five lakes in August 2023. The lakes represent a geographical gradient from central Poland to the Northeast and have different depths, sizes, and trophic statuses. According to performed analyses, the presence of R. raciborskii was recorded in five lakes: Szczęśliwickie (Warsaw), Mikołajskie, Rekąty, Sztynorckie (Masurian Lakeland), and further East, in Pobondzie (Suwałki Lakeland). On the other hand, C. bergii was found in three lakes: Rekąty (Masurian Lakeland), Żabinki, and Pobondzie (Suwałki Lakeland), while S. aphanizomenoides only in Pobondzie (Suwałki Lakeland). Maximum phytoplankton diversity was found in Lake Rekąty, a small and shallow lake mentioned above. The highest phytoplankton biomass was detected in highly eutrophic Lake Suskie, followed by Lake Sztynorckie. In this last lake, which is also strongly eutrophic, the highest biomass of R. raciborskii was found. Cyanophyceae had the highest biovolume and was followed by Chlorophyceae in the entire study. Numerous environmental parameters, including nutrients, were studied, and their relationships with the invasive species and the whole phytoplankton community will be presented. In addition, toxic properties of environmental DNA results from each lake will also be shown. In conclusion, investigated invasive cyanobacterial species were found in a few Northeastern Polish temperate lakes, but the number of individuals was quite low, so the biomass was quite low. It has been observed that the structure of phytoplankton changed based on lakes and environmental parameters.

Keywords: biological invasion, cyanobacteria, cyanotoxins, phytoplankton ecology, sanger sequencing

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102 Genetic Polymorphism and Insilico Study Epitope Block 2 MSP1 Gene of Plasmodium falciparum Isolate Endemic Jayapura

Authors: Arsyam Mawardi, Sony Suhandono, Azzania Fibriani, Fifi Fitriyah Masduki

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Malaria is an infectious disease caused by Plasmodium sp. This disease has a high prevalence in Indonesia, especially in Jayapura. The vaccine that is currently being developed has not been effective in overcoming malaria. This is due to the high polymorphism in the Plasmodium genome especially in areas that encode Plasmodium surface proteins. Merozoite Surface Protein 1 (MSP1) Plasmodium falciparum is a surface protein that plays a role in the invasion process in human erythrocytes through the interaction of Glycophorin A protein receptors and sialic acid in erythrocytes with Reticulocyte Binding Proteins (RBP) and Duffy Adhesion Protein (DAP) ligands in merozoites. MSP1 can be targeted to be a specific antigen and predicted epitope area which will be used for the development of diagnostic and malaria vaccine therapy. MSP1 consists of 17 blocks, each block is dimorphic, and has been marked as the K1 and MAD20 alleles. Exceptions only in block 2, because it has 3 alleles, among others K1, MAD20 and RO33. These polymorphisms cause allelic variations and implicate the severity of patients infected P. falciparum. In addition, polymorphism of MSP1 in Jayapura isolates has not been reported so it is interesting to be further identified and projected as a specific antigen. Therefore, in this study, we analyzed the allele polymorphism as well as detected the MSP1 epitope antigen candidate on block 2 P. falciparum. Clinical samples of selected malaria patients followed the consecutive sampling method, examining malaria parasites with blood preparations on glass objects observed through a microscope. Plasmodium DNA was isolated from the blood of malarial positive patients. The block 2 MSP1 gene was amplified using PCR method and cloned using the pGEM-T easy vector then transformed to TOP'10 E.coli. Positive colonies selection was performed with blue-white screening. The existence of target DNA was confirmed by PCR colonies and DNA sequencing methods. Furthermore, DNA sequence analysis was done through alignment and formation of a phylogenetic tree using MEGA 6 software and insilico analysis using IEDB software to predict epitope candidate for P. falciparum. A total of 15 patient samples have been isolated from Plasmodium DNA. PCR amplification results show the target gene size about ± 1049 bp. The results of MSP1 nucleotide alignment analysis reveal that block 2 MSP1 genes derived from the sample of malarial patients were distributed in four different allele family groups, K1 (7), MAD20 (1), RO33 (0) and MSP1_Jayapura (10) alleles. The most commonly appears of the detected allele is MSP1_Jayapura single allele. There was no significant association between sex variables, age, the density of parasitemia and alel variation (Mann Whitney, U > 0.05), while symptomatic signs have a significant difference as a trigger of detectable allele variation (U < 0.05). In this research, insilico study shows that there is a new epitope antigen candidate from the MSP1_Jayapura allele and it is predicted to be recognized by B cells with 17 amino acid lengths in the amino acid sequence 187 to 203.

Keywords: epitope candidate, insilico analysis, MSP1 P. falciparum, polymorphism

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101 Incorporating Spatial Transcriptome Data into Ligand-Receptor Analyses to Discover Regional Activation in Cells

Authors: Eric Bang

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Interactions between receptors and ligands are crucial for many essential biological processes, including neurotransmission and metabolism. Ligand-receptor analyses that examine cell behavior and interactions often utilize cell type-specific RNA expressions from single-cell RNA sequencing (scRNA-seq) data. Using CellPhoneDB, a public repository consisting of ligands, receptors, and ligand-receptor interactions, the cell-cell interactions were explored in a specific scRNA-seq dataset from kidney tissue and portrayed the results with dot plots and heat maps. Depending on the type of cell, each ligand-receptor pair was aligned with the interacting cell type and calculated the positori probabilities of these associations, with corresponding P values reflecting average expression values between the triads and their significance. Using single-cell data (sample kidney cell references), genes in the dataset were cross-referenced with ones in the existing CellPhoneDB dataset. For example, a gene such as Pleiotrophin (PTN) present in the single-cell data also needed to be present in the CellPhoneDB dataset. Using the single-cell transcriptomics data via slide-seq and reference data, the CellPhoneDB program defines cell types and plots them in different formats, with the two main ones being dot plots and heat map plots. The dot plot displays derived measures of the cell to cell interaction scores and p values. For the dot plot, each row shows a ligand-receptor pair, and each column shows the two interacting cell types. CellPhoneDB defines interactions and interaction levels from the gene expression level, so since the p-value is on a -log10 scale, the larger dots represent more significant interactions. By performing an interaction analysis, a significant interaction was discovered for myeloid and T-cell ligand-receptor pairs, including those between Secreted Phosphoprotein 1 (SPP1) and Fibronectin 1 (FN1), which is consistent with previous findings. It was proposed that an effective protocol would involve a filtration step where cell types would be filtered out, depending on which ligand-receptor pair is activated in that part of the tissue, as well as the incorporation of the CellPhoneDB data in a streamlined workflow pipeline. The filtration step would be in the form of a Python script that expedites the manual process necessary for dataset filtration. Being in Python allows it to be integrated with the CellPhoneDB dataset for future workflow analysis. The manual process involves filtering cell types based on what ligand/receptor pair is activated in kidney cells. One limitation of this would be the fact that some pairings are activated in multiple cells at a time, so the manual manipulation of the data is reflected prior to analysis. Using the filtration script, accurate sorting is incorporated into the CellPhoneDB database rather than waiting until the output is produced and then subsequently applying spatial data. It was envisioned that this would reveal wherein the cell various ligands and receptors are interacting with different cell types, allowing for easier identification of which cells are being impacted and why, for the purpose of disease treatment. The hope is this new computational method utilizing spatially explicit ligand-receptor association data can be used to uncover previously unknown specific interactions within kidney tissue.

Keywords: bioinformatics, Ligands, kidney tissue, receptors, spatial transcriptome

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100 Migratory Diaspora: The Media and the Human Element

Authors: Peter R. Alfieri

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The principal aim of this research and presentation is to give global and personal perspective of the migratory diaspora and how it is perceived by a substantial majority that relies on the media’s portrayal of migratory movements. Since its Greek origins the word “diaspora” has taken on several connotations, but none has surpassed its use in regard to the human element; because since before the dawn of history, man has had to struggle for survival. That survival was a struggle against the elements and other natural enemies, but none as tenacious and relentless as other men. Many have used the term diaspora to describe the spread of certain ethnic groups resulting in new generations in new places; but has the human diaspora been as haphazard as that of spores? The quest for survival has spawned migrations that are not quite that simple, even though it has several similarities to plant spores or dandelion seeds flying throughout the atmosphere. Man kind has constantly migrated in search of food, shelter, and safety. When they were able to find food and shelter, they would inform others who would venture to the new place. Information, whether through word of mouth, written material, or visual communications, has been a moving force in man’s life; and it spurred migrants in their quest for better environments. Today we pride ourselves in being able to communicate instantly with anyone anywhere in the world, and we are privileged to see most of what is happening in the world thanks to the highly developed modern media. Is Media a “wind/force” instrumental in propelling the diaspora throughout the world? The media has been the tool that has incentive many migratory, but unfortunately it is also the means responsible for many misconceptions regarding migrants and their hosts. Has the Media presented an unbiased view of the migrant or has it been the means that generated negative or prejudiced views of the migrant and, perhaps, the host environment? Some examples were easily seen in 19th century the United States where they advertised the following, “Help needed, Irish need not apply”. How do immigrants circumvent latent barriers that are not as obvious as the ones just mentioned? Some immigrants return home and have children that decide to emigrate. It is a perpetual cycle in the search for self-improvement. The stories that are brought back might be inspiration for the new generation of emigrants. Poverty, hunger, and political turmoil spur most migrations. The majority learn from others or through the media about certain destinations that will provide one or several opportunities to improve their existence. Many of those migrants suffer untold hardships to succeed. When they succeed, they provide a great incentive for their children to obtain an education or skill that will insure them a better life. Although the new environment may contribute greatly to a successful career, most immigrants do not forget their own struggle. They see the media’s portrayal of other migrants from all over the globe. Some try to communicate to others the true feelings of despair felt by immigrants, because they are all brothers and sisters in the perennial struggle for a better life. “HOPE” for a better life drives the immigrant toward the unknown and it has helped overcome the obstacles that present themselves challenging every newcomer. Hope and perseverance strengthen the resolve of the migrant in his struggle to survive.

Keywords: media, migration, heath, education, obstacles

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99 CD97 and Its Role in Glioblastoma Stem Cell Self-Renewal

Authors: Niklas Ravn-Boess, Nainita Bhowmick, Takamitsu Hattori, Shohei Koide, Christopher Park, Dimitris Placantonakis

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Background: Glioblastoma (GBM) is the most common and deadly primary brain malignancy in adults. Tumor propagation, brain invasion, and resistance to therapy critically depend on GBM stem-like cells (GSCs); however, the mechanisms that regulate GSC self-renewal are incompletely understood. Given the aggressiveness and poor prognosis of GBM, it is imperative to find biomarkers that could also translate into novel drug targets. Along these lines, we have identified a cell surface antigen, CD97 (ADGRE5), an adhesion G protein-coupled receptor (GPCR), that is expressed on GBM cells but is absent from non-neoplastic brain tissue. CD97 has been shown to promote invasiveness, angiogenesis, and migration in several human cancers, but its frequency of expression and functional role in regulating GBM growth and survival, and its potential as a therapeutic target has not been investigated. Design: We assessed CD97 mRNA and protein expression in patient derived GBM samples and cell lines using publicly available RNA-sequencing datasets and flow cytometry, respectively. To assess CD97 function, we generated shRNA lentiviral constructs that target a sequence in the CD97 extracellular domain (ECD). A scrambled shRNA (scr) with no predicted targets in the genome was used as a control. We evaluated CD97 shRNA lentivirally transduced GBM cells for Ki67, Annexin V, and DAPI. We also tested CD97 KD cells for their ability to self-renew using clonogenic tumorsphere formation assays. Further, we utilized synthetic Abs (sAbs) generated against the ECD of CD97 to test for potential antitumor effects using patient-derived GBM cell lines. Results: CD97 mRNA expression was expressed at high levels in all GBM samples available in the TCGA cohort. We found high levels of surface CD97 protein expression in 6/6 patient-derived GBM cell cultures, but not human neural stem cells. Flow cytometry confirmed downregulation of CD97 in CD97 shRNA lentivirally transduced cells. CD97 KD induced a significant reduction in cell growth in 3 independent GBM cell lines representing mesenchymal and proneural subtypes, which was accompanied by reduced (~20%) Ki67 staining and increased (~30%) apoptosis. Incubation of GBM cells with sAbs (20 ug/ ml) against the ECD of CD97 for 3 days induced GSC differentiation, as determined by the expression of GFAP and Tubulin. Using three unique GBM patient derived cultures, we found that CD97 KD attenuated the ability of GBM cells to initiate sphere formation by over 300 fold, consistent with an impairment in GSC self-renewal. Conclusion: Loss of CD97 expression in patient-derived GBM cells markedly decreases proliferation, induces cell death, and reduces tumorsphere formation. sAbs against the ECD of CD97 reduce tumorsphere formation, recapitulating the phenotype of CD97 KD, suggesting that sAbs that inhibit CD97 function exhibit anti-tumor activity. Collectively, these findings indicate that CD97 is necessary for the proliferation and survival of human GBM cells and identify CD97 as a promising therapeutically targetable vulnerability in GBM.

Keywords: adhesion GPCR, CD97, GBM stem cell, glioblastoma

Procedia PDF Downloads 133
98 Evaluating Impact of Teacher Professional Development Program on Students’ Learning

Authors: S. C. Lin, W. W. Cheng, M. S. Wu

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This study attempted to investigate the connection between teacher professional development program and students’ Learning. This study took Readers’ Theater Teaching Program (RTTP) for professional development as an example to inquiry how participants apply their new knowledge and skills learned from RTTP to their teaching practice and how the impact influence students learning. The goals of the RTTP included: 1) to enhance teachers RT content knowledge; 2) to implement RT instruction in teachers’ classrooms in response to their professional development. 2) to improve students’ ability of reading fluency in professional development teachers’ classrooms. This study was a two-year project. The researchers applied mixed methods to conduct this study including qualitative inquiry and one-group pretest-posttest experimental design. In the first year, this study focused on designing and implementing RTTP and evaluating participants’ satisfaction of RTTP, what they learned and how they applied it to design their English reading curriculum. In the second year, the study adopted quasi-experimental design approach and evaluated how participants RT instruction influenced their students’ learning, including English knowledge, skill, and attitudes. The participants in this study composed two junior high school English teachers and their students. Data were collected from a number of different sources including teaching observation, semi-structured interviews, teaching diary, teachers’ professional development portfolio, Pre/post RT content knowledge tests, teacher survey, and students’ reading fluency tests. To analyze the data, both qualitative and quantitative data analysis were used. Qualitative data analysis included three stages: organizing data, coding data, and analyzing and interpreting data. Quantitative data analysis included descriptive analysis. The results indicated that average percentage of correct on pre-tests in RT content knowledge assessment was 40.75% with two teachers ranging in prior knowledge from 35% to 46% in specific RT content. Post-test RT content scores ranged from 70% to 82% correct with an average score of 76.50%. That gives teachers an average gain of 35.75% in overall content knowledge as measured by these pre/post exams. Teachers’ pre-test scores were lowest in script writing and highest in performing. Script writing was also the content area that showed the highest gains in content knowledge. Moreover, participants hold a positive attitude toward RTTP. They recommended that the approach of professional learning community, which was applied in RTTP was benefit to their professional development. Participants also applied the new skills and knowledge which they learned from RTTP to their practices. The evidences from this study indicated that RT English instruction significantly influenced students’ reading fluency and classroom climate. The result indicated that all of the experimental group students had a big progress in reading fluency after RT instruction. The study also found out several obstacles. Suggestions were also made.

Keywords: teacher’s professional development, program evaluation, readers’ theater, english reading instruction, english reading fluency

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97 Defective Autophagy Disturbs Neural Migration and Network Activity in hiPSC-Derived Cockayne Syndrome B Disease Models

Authors: Julia Kapr, Andrea Rossi, Haribaskar Ramachandran, Marius Pollet, Ilka Egger, Selina Dangeleit, Katharina Koch, Jean Krutmann, Ellen Fritsche

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It is widely acknowledged that animal models do not always represent human disease. Especially human brain development is difficult to model in animals due to a variety of structural and functional species-specificities. This causes significant discrepancies between predicted and apparent drug efficacies in clinical trials and their subsequent failure. Emerging alternatives based on 3D in vitro approaches, such as human brain spheres or organoids, may in the future reduce and ultimately replace animal models. Here, we present a human induced pluripotent stem cell (hiPSC)-based 3D neural in a vitro disease model for the Cockayne Syndrome B (CSB). CSB is a rare hereditary disease and is accompanied by severe neurologic defects, such as microcephaly, ataxia and intellectual disability, with currently no treatment options. Therefore, the aim of this study is to investigate the molecular and cellular defects found in neural hiPSC-derived CSB models. Understanding the underlying pathology of CSB enables the development of treatment options. The two CSB models used in this study comprise a patient-derived hiPSC line and its isogenic control as well as a CSB-deficient cell line based on a healthy hiPSC line (IMR90-4) background thereby excluding genetic background-related effects. Neurally induced and differentiated brain sphere cultures were characterized via RNA Sequencing, western blot (WB), immunocytochemistry (ICC) and multielectrode arrays (MEAs). CSB-deficiency leads to an altered gene expression of markers for autophagy, focal adhesion and neural network formation. Cell migration was significantly reduced and electrical activity was significantly increased in the disease cell lines. These data hint that the cellular pathologies is possibly underlying CSB. By induction of autophagy, the migration phenotype could be partially rescued, suggesting a crucial role of disturbed autophagy in defective neural migration of the disease lines. Altered autophagy may also lead to inefficient mitophagy. Accordingly, disease cell lines were shown to have a lower mitochondrial base activity and a higher susceptibility to mitochondrial stress induced by rotenone. Since mitochondria play an important role in neurotransmitter cycling, we suggest that defective mitochondria may lead to altered electrical activity in the disease cell lines. Failure to clear the defective mitochondria by mitophagy and thus missing initiation cues for new mitochondrial production could potentiate this problem. With our data, we aim at establishing a disease adverse outcome pathway (AOP), thereby adding to the in-depth understanding of this multi-faced disorder and subsequently contributing to alternative drug development.

Keywords: autophagy, disease modeling, in vitro, pluripotent stem cells

Procedia PDF Downloads 119