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

Search results for: sequencing skill

396 Enhancing Higher Education Teaching and Learning Processes: Examining How Lecturer Evaluation Make a Difference

Authors: Daniel Asiamah Ameyaw

Abstract:

This research attempts to investigate how lecturer evaluation makes a difference in enhancing higher education teaching and learning processes. The research questions to guide this research work states first as, “What are the perspectives on the difference made by evaluating academic teachers in order to enhance higher education teaching and learning processes?” and second, “What are the implications of the findings for Policy and Practice?” Data for this research was collected mainly through interviewing and partly documents review. Data analysis was conducted under the framework of grounded theory. The findings showed that for individual lecturer level, lecturer evaluation provides a continuous improvement of teaching strategies, and serves as source of data for research on teaching. At the individual student level, it enhances students learning process; serving as source of information for course selection by students; and by making students feel recognised in the educational process. At the institutional level, it noted that lecturer evaluation is useful in personnel and management decision making; it assures stakeholders of quality teaching and learning by setting up standards for lecturers; and it enables institutions to identify skill requirement and needs as a basis for organising workshops. Lecturer evaluation is useful at national level in terms of guaranteeing the competencies of graduates who then provide the needed manpower requirement of the nation. Besides, it mentioned that resource allocation to higher educational institution is based largely on quality of the programmes being run by the institution. The researcher concluded, that the findings have implications for policy and practice, therefore, higher education managers are expected to ensure that policy is implemented as planned by policy-makers so that the objectives can successfully be achieved.

Keywords: academic quality, higher education, lecturer evaluation, teaching and learning processes

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395 Bean in Turkey: Characterization, Inter Gene Pool Hybridization Events, Breeding, Utilizations

Authors: Faheem Shahzad Baloch, Muhammad Azhar Nadeem, Muhammad Amjad Nawaz, Ephrem Habyarimana, Gonul Comertpay, Tolga Karakoy, Rustu Hatipoglu, Mehmet Zahit Yeken, Vahdettin Ciftci

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Turkey is considered a bridge between Europe, Asia, and Africa and possibly played an important role in the distribution of many crops including common bean. Hundreds of common bean landraces can be found in Turkey, particularly in farmers’ fields, and they consistently contribute to the overall production. To investigate the existing genetic diversity and hybridization events between the Andean and Mesoamerican gene pools in the Turkish common bean, 188 common bean accessions (182 landraces and 6 modern cultivars as controls) were collected from 19 different Turkish geographic regions. These accessions were characterized using phenotypic data (growth habit and seed weight), geographic provenance, 12557 high-quality whole-genome DArTseq markers, and 3767 novel DArTseq loci were also identified. The clustering algorithms resolved the Turkish common bean landrace germplasm into the two recognized gene pools, the Mesoamerican and Andean gene pools. Hybridization events were observed in both gene pools (14.36% of the accessions) but mostly in the Mesoamerican (7.97% of the accessions), and was low relative to previous European studies. The lower level of hybridization witnessed the existence of Turkish common bean germplasm in its original form as compared to Europe. Mesoamerican gene pool reflected a higher level of diversity, while the Andean gene pool was predominant (56.91% of the accessions), but genetically less diverse and phenotypically more pure, reflecting farmers greater preference for the Andean gene pool. We also found some genetically distinct landraces and overall, a meaningful level of genetic variability which can be used by the scientific community in breeding efforts to develop superior common bean strains.

Keywords: bean germplasm, DArTseq markers, genotyping by sequencing, Turkey, whole genome diversity

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394 Supporting Regulation and Shared Attention to Facilitate the Foundations for Development of Children and Adolescents with Complex Individual Profiles

Authors: Patsy Tan, Dana Baltutis

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This presentation demonstrates the effectiveness of music therapy in co-treatment with speech pathology and occupational therapy as an innovative way when working with children and adolescents with complex individual differences to facilitate communication, emotional, motor and social skills development. Each child with special needs and their carer has an individual profile which encompasses their visual-spatial, auditory, language, learning, mental health, family dynamic, sensory-motor, motor planning and sequencing profiles. The most common issues among children with special needs, especially those diagnosed with Autism Spectrum Disorder, are in the areas of regulation, communication, and social-emotional development. The ability of children living with challenges to communicate and use language and understand verbal and non-verbal information, as well as move their bodies to explore and interact with their environments in social situations, depends on the children being regulated both internally and externally and trusting their communication partners and understanding what is happening in the moment. For carers, it is about understanding the tempo, rhythm, pacing, and timing of their own individual profile, as well as the profile of the child they are interacting with, and how these can sync together. In this study, music therapy is used in co-treatment sessions with a speech pathologist and/or an occupational therapist using the DIRFloortime approach to facilitate the regulation, attention, engagement, reciprocity and social-emotional capacities of children presenting with complex individual differences. Documented changes in 10 domains of children’s development over a 12-month period using the Individual Music Therapy Assessment Profile (IMTAP) were observed. Children were assessed biannually, and results show significant improvements in the social-emotional, musicality and receptive language domains indicating that co-treatment with a music therapist using the DIRFloortime framework is highly effective. This presentation will highlight strategies that facilitate regulation, social-emotional and communication development for children and adolescents with complex individual profiles.

Keywords: communication, shared attention, regulation, social emotional

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393 Confidence Levels among UK Emergency Medicine Doctors in Performing Emergency Lateral Canthotomy: Should it be a Key Skill in the ED

Authors: Mohanad Moustafa, Julia Sieberer, Rhys Davies

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Background: Orbital compartment syndrome (OCS) is a sight-threatening Ophthalmologic emergency caused by rapidly increasing intraorbital pressure. It is usually caused by a retrobulbar hemorrhage as a result of trauma. If not treated in a timely manner, permanent vision loss can occur. Lateral canthotomy and cantholysis are minor procedures that can be performed bedside with equipment available in the emergency department. The aim of the procedure is to release the attachments between the suspensory ligaments of the eye and the bony orbital wall, leading to a decrease in intraorbital pressure and preventing irreversible loss of vision. As most Ophthalmologists across the UK provide non-resident on-call service, this may lead to a delay in the treatment of OCS and stresses the need for Emergency medical staff to be able to provide this sight-saving procedure independently. Aim: To survey current training, experience, and confidence levels among Emergency Medicine doctors in performing emergency lateral canthotomy and to establish whether these variables change the following teaching from experienced ophthalmologists. RESULTS: Most EM registrars had little to no experience in performing lateral canthotomy and cantholysis. The majority of them showed a significant increase in their confidence to perform the procedure following ophthalmic-led teaching. The survey also showed that the registrars felt such training should be added to/part of the EM curriculum. Conclusion: The involvement of Ophthalmologists in the teaching of EM doctors to recognise and treat OCS independently may prevent delays in treatment and reduce the risk of permanent sight loss. This project showed potential in improving patient care and will lead to a National Survey of EM doctors across the UK.

Keywords: lateral canthotomy, retrobulbar hemorrhage, Ophthalmology, orbital compartment syndrome, sight loss, blindness

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392 Using Multiomic Plasma Profiling From Liquid Biopsies to Identify Potential Signatures for Disease Diagnostics in Late-Stage Non-small Cell Lung Cancer (NSCLC) in Trinidad and Tobago

Authors: Nicole Ramlachan, Samuel Mark West

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Lung cancer is the leading cause of cancer-associated deaths in North America, with the vast majority being non-small cell lung cancer (NSCLC), with a five-year survival rate of only 24%. Non-invasive discovery of biomarkers associated with early-diagnosis of NSCLC can enable precision oncology efforts using liquid biopsy-based multiomics profiling of plasma. Although tissue biopsies are currently the gold standard for tumor profiling, this method presents many limitations since these are invasive, risky, and sometimes hard to obtain as well as only giving a limited tumor profile. Blood-based tests provides a less-invasive, more robust approach to interrogate both tumor- and non-tumor-derived signals. We intend to examine 30 stage III-IV NSCLC patients pre-surgery and collect plasma samples.Cell-free DNA (cfDNA) will be extracted from plasma, and next-generation sequencing (NGS) performed. Through the analysis of tumor-specific alterations, including single nucleotide variants (SNVs), insertions, deletions, copy number variations (CNVs), and methylation alterations, we intend to identify tumor-derived DNA—ctDNA among the total pool of cfDNA. This would generate data to be used as an accurate form of cancer genotyping for diagnostic purposes. Using liquid biopsies offer opportunities to improve the surveillance of cancer patients during treatment and would supplement current diagnosis and tumor profiling strategies previously not readily available in Trinidad and Tobago. It would be useful and advantageous to use this in diagnosis and tumour profiling as well as to monitor cancer patients, providing early information regarding disease evolution and treatment efficacy, and reorient treatment strategies in, timethereby improving clinical oncology outcomes.

Keywords: genomics, multiomics, clinical genetics, genotyping, oncology, diagnostics

Procedia PDF Downloads 157
391 Clients’ Priorities in Delivery of Green Projects: South African Perspective

Authors: C. Mothobiso, D. Root

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Purpose: This study attempts to identify the clients’ main priorities when delivering green projects. The aim is to compare if the clients have the same interest that are similar in delivery of convectional buildings as compared to green buildings. The main purpose is to find why other clients are investing in green buildings while others are reluctant and adopting green building at a slow pace. Design/methodology/approach: A sample of construction professional accredited by the Green Building Council of South Africa (GBCSA) was sent a questionnaire to participate in the research. Since GBSCSA accredited professionals have knowledge and experience about the green buildings, they are chosen as the sample. The research is qualitative because it evaluates the perceptions and knowledge around the subject matter. Research limitations: The research focuses only on the South African construction clients. Findings: Findings reveal that private clients invest more on green buildings as compared to government and parastatal entities. Private clients prioritise on maximising returns on investments and they mainly invest on buildings that save energies and have low life cycle costs. Private clients are perceived to be more knowledgeable about the benefits of green building project as compared to government and Parastatals clients. Shortage of expertise and managerial skill leads to low adaptation of green buildings in the government and parastatal projects. Other factors, which seem to disintegrate the adoption of green buildings, are the readiness of supply chain within the industry and inappropriate procurements strategies adopted by clients. The evaluation of the clients’ priorities will enable the design team to come up with innovative ways to approach the design process so that clients’ priorities and needs are identified and met. Practical implications: The findings are indicating that clients’ needs and priorities have a huge impact on the delivery of the project in terms of time, quality and cost of the project.

Keywords: construction clients, design team, green construction and project deliver

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390 Analysis of Farm Management Skills in Broiler Poultry Producers in Botswana

Authors: Som Pal Baliyan

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The purpose of this quantitative study was to analyze farm management skills in broiler poultryproducers in Botswana. The study adopted a descriptive and correlation research design. The population of the study was the poultry farm operators who had been in broiler poultry farming at least for two years. Based on the information from literature, a questionnaire was constructed for data collection on seven areas of farm management skills namely; planning skills, accounting and financial management skills, production management skills, product procurement and marketing skills, decision making skills, risk management skills, and specific technical skills. The validity and reliability of the questionnaire were accomplished by a panel of experts and by calculating the Cronbach’s alpha coefficient, respectively. Data were collected through a survey of 60 randomly sampled poultry farm operators in Botswana. Data were analyzed through descriptive statistical tools whereby the level of farm management skills were determined by calculating means and standard deviations of the management skills among the broiler producers. The level of farm management skills in broilers producers was discussed. All the seven farm management skills were ranked based on their calculated means. The specific technical skills and risk management skills were the highest and the lowest ranked farm management skills, respectively.Findings revealed that the broiler producers had skills above the average level only in specific technical skills whereas the skill levels in the remaining six farm management skills under study were found below the average level. This prevailing low level of farm management skills can be justified asthe cause of failure or poor performance of the broiler poultry farms in Botswana. Therefore, in order to improve the efficiency and productivityin broiler production in the country, it was recommended that the broiler poultry producers should be adequately trained in areas of planning skills, financial management skills, production management skills, product procurement and marketing skills, decision making skills and risk management skills.

Keywords: poultry production, broiler production, management skills, levels of skills

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389 Milk Yield and Fingerprinting of Beta-Casein Precursor (CSN2) Gene in Some Saudi Camel Breeds

Authors: Amr A. El Hanafy, Yasser M. Saad, Saleh A. Alkarim, Hussein A. Almehdar, Elrashdy M. Redwan

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Camels are substantial providers of transport, milk, sport, meat, shelter, fuel, security and capital in many countries, particularly Saudi Arabia. Identification of animal breeds has progressed rapidly during the last decade. Advanced molecular techniques are playing a significant role in breeding or strain protection laws. On the other hand, fingerprinting of some molecular markers related to some productive traits in farm animals represents most important studies to our knowledge, which aim to conserve these local genetic resources, and to the genetic improvement of such local breeds by selective programs depending on gene markers. Milk records were taken two days in each week from female camels of Majahem, Safara, Wathaha, and Hamara breeds, respectively from different private farms in northern Jeddah, Riyadh and Alwagh governorates and average weekly yields were calculated. DNA sequencing for CSN2 gene was used for evaluating the genetic variations and calculating the genetic distance values among four Saudi camel populations which are Hamra(R), Safra(Y), Wadha(W) and Majaheim(M). In addition, this marker was analyzed for reconstructing the Neighbor joining tree among evaluating camel breeds. In respect to milk yield during winter season, result indicated that average weekly milk yield of Safara camel breed (30.05 Kg/week) is significantly (p < 0.05) lower than the other 3 breeds which ranged from 39.68 for Hamara to 42.42 Kg/week for Majahem, while there are not significant differences between these three breeds. The Neighbor Joining analysis that re-constructed based on DNA variations showed that samples are clustered into two unique clades. The first clade includes Y (from Y4 to Y18) and M (from M1, to M9). On the other hand, the second cluster is including all R (from R1 to R6) and W (from W1 to W6). The genetic distance values were equal 0.0068 (between the groups M&Y and R&W) and equal 0 (within each group).

Keywords: milk yield, beta-casein precursor (CSN2), Saudi camel, molecular markers

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388 Practical Modelling of RC Structural Walls under Monotonic and Cyclic Loading

Authors: Reza E. Sedgh, Rajesh P. Dhakal

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Shear walls have been used extensively as the main lateral force resisting systems in multi-storey buildings. The recent development in performance based design urges practicing engineers to conduct nonlinear static or dynamic analysis to evaluate seismic performance of multi-storey shear wall buildings by employing distinct analytical models suggested in the literature. For practical purpose, application of macroscopic models to simulate the global and local nonlinear behavior of structural walls outweighs the microscopic models. The skill level, computational time and limited access to RC specialized finite element packages prevents the general application of this method in performance based design or assessment of multi-storey shear wall buildings in design offices. Hence, this paper organized to verify capability of nonlinear shell element in commercially available package (Sap2000) in simulating results of some specimens under monotonic and cyclic loads with very oversimplified available cyclic material laws in the analytical tool. The selection of constitutive models, the determination of related parameters of the constituent material and appropriate nonlinear shear model are presented in detail. Adoption of proposed simple model demonstrated that the predicted results follow the overall trend of experimental force-displacement curve. Although, prediction of ultimate strength and the overall shape of hysteresis model agreed to some extent with experiment, the ultimate displacement(significant strength degradation point) prediction remains challenging in some cases.

Keywords: analytical model, nonlinear shell element, structural wall, shear behavior

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387 English Language Teaching Graduate Students' Use of Discussion Moves in Research Articles

Authors: Gamzegul Koca, Evrim Eveyik-Aydin

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Genre and discipline-specific knowledge of academic discourse in writing has long been acknowledged as being a core skill to achieve formidable tasks that are expected of graduate students in academic settings. Genre analysis approaches can be adopted to unveil the challenges encountered in these tasks to be able to take instructional actions addressing the aspects of graduate writing that need improvement. In an attempt to find genre-specific academic writing needs of Turkish students enrolled in a graduate program in ELT, this study examines the rhetorical structure of discussion sections of research articles written during the course load stage of their graduate studies. The 35.437-word specialized corpus of graduate papers compiled for the purpose of the study includes discussions of 58 unpublished reports of empirical studies, 31 written in MA courses and 27 in Ph.D. courses by a total of 44 graduate students. The study does sentence-based move structure analysis using the framework developed by Eveyik-Aydın, Karabacak and Akyel in a corpus-based study that analyzed the discussion moves of expert writers in published articles in ELT journals indexed by Social Sciences Citation. The coding of 1577 sentences by three graders using this framework revealed that while the graduate papers included the same moves used in published articles, the rhetorical structure of MA and Ph.D. papers showed considerable differences in terms of the frequency of occurrence of main discussion moves, including interpretation of the results and drawing implications. The implications of these findings will be discussed with respect to the needs of graduate writers and the expectations of discourse community.

Keywords: discussion moves, genre-specific rhetorical structure, move analysis, research articles, the specialized corpus of graduate papers

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386 Neuropsychological Disabilities in Executive Functions and Visuospatial Skills of Juvenile Offenders in a Half-Open Program in Santiago De Chile

Authors: Gabriel Sepulveda Navarro

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Traditional interventions for young offenders are necessary but not sufficient to tackle the multiple causes of juvenile crime. For instance, interventions offered to young offenders often are verbally mediated and dialogue based, requiring important metacognitive abilities as well as abstract thinking, assuming average performance in a wide variety of skills. It seems necessary to assess a broader set of abilities and functions in order to increase the efficiency of interventions while addressing offending. In order to clarify these assumptions, Stroop Test, as well as Rey-Osterrieth Complex Figure Test were applied to juvenile offenders tried and sentenced for violent crimes in Santiago de Chile. A random sample was drawn from La Cisterna Half-Open Program, consisting of 50 young males between 18 and 24 years old, residing in different districts of Santiago de Chile. The analysis of results suggests a disproportionately elevated incidence of impairments in executive functions and visuospatial skills. As an outcome, over 40% of the sample shows a significant low performance in both assessments, exceeding four times the same prevalence rates among young people in the general population. While executive functions entail working memory (being able to keep information and use it in some way), cognitive flexibility (to think about something in more than one way) and inhibitory control (being able to self-control, ignore distractions and delay immediate gratification), visuospatial skills permit to orientate and organize a planned conduct. All of these abilities are fundamental to the skill of avoiding violent behaviour and abiding by social rules. Understanding the relevance of neurodevelopmental impairments in the onset of violent and criminal behaviour, as well as recidivism, eventually may guide the deployment of a more comprehensive assessment and treatment for juvenile offenders.

Keywords: executive functions, half-open program, juvenile offenders, neurodisabilities, visuospatial skills

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385 The Effect of PETTLEP Imagery on Equestrian Jumping Tasks

Authors: Nurwina Anuar, Aswad Anuar

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Imagery is a popular mental technique used by athletes and coaches to improve learning and performance. It has been widely investigated and beneficial in the sports context. However, the imagery application in equestrian sport has been understudied. Thus, the effectiveness of imagery should encompass the application in the equestrian sport to ensure its application covert all sports. Unlike most sports (e.g., football, badminton, tennis, ski) which are both mental and physical are dependent solely upon human decision and response, equestrian sports involves the interaction of human-horse collaboration to success in the equestrian tasks. This study aims to investigate the effect of PETTLEP imagery on equestrian jumping tasks, motivation and imagery ability. It was hypothesized that the use of PETTLEP imagery intervention will significantly increase in the skill equestrian jumping tasks. It was also hypothesized that riders’ imagery ability and motivation will increase across phases. The participants were skilled riders with less to no imagery experience. A single-subject ABA design was employed. The study was occurred over five week’s period at Universiti Teknologi Malaysia Equestrian Park. Imagery ability was measured using the Sport Imagery Assessment Questionnaires (SIAQ), the motivational measured based on the Motivational imagery ability measure for Sport (MIAMS). The effectiveness of the PETTLEP imagery intervention on show jumping tasks were evaluated by the professional equine rider on the observational scale. Results demonstrated the improvement on all equestrian jumping tasks for the most participants from baseline to intervention. Result shows the improvement on imagery ability and participants’ motivations after the PETTLEP imagery intervention. Implication of the present study include underlining the impact of PETTLEP imagery on equestrian jumping tasks. The result extends the previous research on the effectiveness of PETTLEP imagery in the sports context that involves interaction and collaboration between human and horse.

Keywords: PETTLEP imagery, imagery ability, equestrian, equestrian jumping tasks

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384 Speech Anxiety in Higher Education Students-Retention of an Ancestral Trait: A Study into the Students' Perspective of Communication Anxiety with Suggestions on How to Minimise Student Distress

Authors: Paul D. Facey, Claire Morgan

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Speech anxiety is thought to be deep-seated within the human evolutionary lineage.As a result, almost all people display high levels of anxiety when asked to communicate in front of an audience.However, proficiency in oral communication is considered as an essential skill for a graduate career and significant emphasis is placed on developing these skills in many degree programs.Because of this, many degree schemes incorporate some form of assessed dialogic presentation. Yet, a student’s anxiety over public speaking, especially if severe, can be so great that at worst it can cause the student to withdraw from their study. This study investigated how students perceive their own levels of anxiety when faced with public speaking using the Personal Report of Public Speaking Anxiety (PRPSA) questionnaire developed by McCroskey. Additionally, students were asked to provide examples of adjustments that could be implemented that they felt would alleviate some/all of their anxiety. The results of the study indicated that the majority of the students experienced a moderate level of anxiety. However, further analysis showed that of those who were in the moderate anxiety’ group, 43% fell into the higher range suggesting that overall more students experience higher levels of anxiety when faced with public speaking than maybe first envisaged. Thus, it is essential that steps are taken to address student anxiety in order that students engage with presentations, are motivated and encouraged and do not avoid such assignments. The feedback from our students indicated a need to implement systematic desensitization programs where students learn to overcome their anxiety through a series of sessions that gradually increase their anxiety levels. Furthermore, these sessions should be run in parallel with skills sessions in order for students to be better prepared and allow self-reflection and self-analysis.This study highlights the paucity of these sessions on many degree schemes and suggests that they should form an integral part of a students’ early academic learning.

Keywords: student anxiety, communication anxiety, public speaking, higher education, desensitisation

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383 Characterization of Defense-Related Genes and Metabolite Profiling in Oil Palm Elaeis guineensis during Interaction with Ganoderma boninense

Authors: Mohammad Nazri Abdul Bahari, Nurshafika Mohd Sakeh, Siti Nor Akmar Abdullah

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Basal stem rot (BSR) is the most devastating disease in oil palm. Among the oil palm pathogenic fungi, the most prevalent and virulent species associated with BSR is Ganoderma boninense. Early detection of G. boninense attack in oil palm wherein physical symptoms has not yet appeared can offer opportunities to prevent the spread of the necrotrophic fungus. However, poor understanding of molecular defense responses and roles of antifungal metabolites in oil palm against G. boninense has complicated the resolving measures. Hence, characterization of defense-related molecular responses and production of antifungal compounds during early interaction with G. boninense is of utmost important. Four month-old oil palm (Elaeis guineensis) seedlings were artificially infected with G. boninense-inoculated rubber wood block via sitting technique. RNA of samples were extracted from roots and leaves tissues at 0, 3, 7 and 11 days post inoculation (d.p.i) followed with sequencing using RNA-Seq method. Differentially-expressed genes (DEGs) of oil palm-G. boninense interaction were identified, while changes in metabolite profile will be scrutinized related to the DEGs. The RNA-Seq data generated a total of 113,829,376 and 313,293,229 paired-end clean reads from untreated (0 d.p.i) and treated (3, 7, 11 d.p.i) samples respectively, each with two biological replicates. The paired-end reads were mapped to Elaeis guineensis reference genome to screen out non-oil palm genes and subsequently generated 74,794 coding sequences. DEG analysis of phytohormone biosynthetic genes in oil palm roots revealed that at p-value ≤ 0.01, ethylene and jasmonic acid may act in antagonistic manner with salicylic acid to coordinate defense response at early interaction with G. boninense. Findings on metabolite profiling of G. boninense-infected oil palm roots and leaves are hoped to explain the defense-related compounds elicited by Elaeis guineensis in response to G. boninense colonization. The study aims to shed light on molecular defense response of oil palm at early interaction with G. boninense and promote prevention measures against Ganoderma infection.

Keywords: Ganoderma boninense, metabolites, phytohormones, RNA-Seq

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382 Extracellular Production of the Oncolytic Enzyme, Glutaminase Free L-Asparaginase, from Newly Isolated Streptomyces Olivaceus NEAE-119: Optimization of Culture Conditions Using Response Surface Methodology

Authors: Noura El-Ahmady El-Naggar

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Among the antitumour drugs, bacterial enzyme L-asparaginase has been employed as the most effective chemotherapeutic agent in pediatric oncotherapy especially for acute lymphoblastic leukemia. Glutaminase free L-asparaginase producing actinomycetes were isolated from soil samples collected from Egypt. Among them, a potential culture, strain NEAE-119, was selected and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as Streptomyces olivaceus NEAE-119 and sequencing product(1509 bp) was deposited in the GenBank database under accession number KJ200342. The optimization of different process parameters for L-asparaginase production by Streptomyces olivaceus NEAE-119 using Plackett–Burman experimental design and response surface methodology was carried out. Fifteen nutritional variables (temperature, pH, incubation time, inoculum size, inoculum age, agitation speed, dextrose, starch, L-asparagine, KNO3, yeast extract, K2HPO4, MgSO4.7H2O, NaCl and FeSO4. 7H2O) were screened using Plackett–Burman experimental design. The most positive significant independent variables affecting enzyme production (temperature, inoculum age and agitation speed) were further optimized by the central composite face-centered design -response surface methodology. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase in the culture filtrate of Streptomyces olivaceus NEAE-119: Dextrose 3g, starch 20g, L-asparagine 10g, KNO3 1g, K2HPO4 1g, MgSO4.7H2O 0.1g, NaCl 0.1g, pH 7, temperature 37°C, agitation speed 200 rpm/min, inoculum size 4%, v/v, inoculum age 72 h and fermentation period 5 days.

Keywords: Streptomyces olivaceus NEAE-119, glutaminase free L-asparaginase, production, Plackett-Burman design, central composite face-centered design, 16S rRNA, scanning electron microscope

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381 The Molecular Rationale for Steroid Based Therapy of Leukemia: Diagnostic and Therapeutic Implications

Authors: Eitan Yefenof

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Glucocorticoid (GC) hormones, e.g. Dexamethasone and Prednisone, are widely used in the therapy of leukemia and lymphoma owing to their apoptogenic effect on lymphoid cells. However, the emergence of GC resistant cells during therapy is a major cause for treatment failure, urging the need for novel strategies that maintain leukemia sensitivity to the pro-apoptotic activity of GCs. GCs act by binding to the GC receptor (GR), which, in its inactive state, is sequestered in the cytosol by a multi-subunit complex of heat shock proteins. Upon ligand binding, the complex dissociates, allowing GR activation and translocation to the nucleus, where it regulates transcription of multiple genes. We demonstrated that in addition to gene expression, GR also regulates microRNA (miR) expression. Deep-sequencing analysis revealed 14 miRs that are regulated in GC-sensitive but resistant leukemias upon treatment with GC. GC up-regulates miR-103, miR-15~16 and miR-30e/d, while down-regulates miR-17, mir-18a, miR-19a, miR-19b, miR-20a and miR-92a (members of the miR-17∼92a multi-cistron). Upon transfection, miR-103 confers GC apoptotic sensitivity to otherwise GC-resistant cell. Furthermore, knocking down miR-103 expression reduces the GC apoptotic response of sensitive cells. miR-103 abrogates c-Myc expression, an oncogenic transcription factor which is deregulated in many cancers. In addition, miR-103 up-regulates Bim, a pro-apoptotic protein crucial for GC-induced death. Activated glycogen synthase kinase 3 (GSK3) is also crucial for GC-induced apoptosis. GSK3 is active in GC-sensitive but not in GC-resistant cells. We found that GSK3 associates with the GR multi-subunit complex. Upon GC exposure, it dissociates from the GR and interacts with Bim to enable activation of the mitochondrial apoptosis pathway. miR-103 mediated c-Myc ablation is followed by down-regulation of the multi-cistron miR-17~92a, in particular miR-18a and miR-20a. miR-18a targets GR for degradation whereas miR-20a targets Bim degradation. Hence, miR-103 acts, in concert with Bim and GR, as a "tumor suppressor" that leads to reduced proliferation, cell-cycle arrest and cell death. We suggest that miR-103 can provide a diagnostic tool that predicts the sensitivity of leukemia to GC based therapy. Furthermore, exosomal delivery of miR-103 or up-regulation of the endogenous miR-103 could confer apoptotic sensitivity to resistant cells at the outset, thus becoming a useful therapeutic tool combined with GCs.

Keywords: apoptosis, leukemia, micro-RNA, steroids

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380 A Nucleic Acid Extraction Method for High-Viscosity Floricultural Samples

Authors: Harunori Kawabe, Hideyuki Aoshima, Koji Murakami, Minoru Kawakami, Yuka Nakano, David D. Ordinario, C. W. Crawford, Iri Sato-Baran

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With the recent advances in gene editing technologies allowing the rewriting of genetic sequences, additional market growth in the global floriculture market beyond previous trends is anticipated through increasingly sophisticated plant breeding techniques. As a prerequisite for gene editing, the gene sequence of the target plant must first be identified. This necessitates the genetic analysis of plants with unknown gene sequences, the extraction of RNA, and comprehensive expression analysis. Consequently, a technology capable of consistently and effectively extracting high-purity DNA and RNA from plants is of paramount importance. Although model plants, such as Arabidopsis and tobacco, have established methods for DNA and RNA extraction, floricultural species such as roses present unique challenges. Different techniques to extract DNA and RNA from various floricultural species were investigated. Upon sampling and grinding the petals of several floricultural species, it was observed that nucleic acid extraction from the ground petal solutions of low viscosity was straightforward; solutions of high viscosity presented a significant challenge. It is postulated that the presence of substantial quantities of polysaccharides and polyphenols in the plant tissue was responsible for the inhibition of nucleic acid extraction. Consequently, attempts were made to extract high-purity DNA and RNA by improving the CTAB method and combining it with commercially available nucleic acid extraction kits. The quality of the total extracted DNA and RNA was evaluated using standard methods. Finally, the effectiveness of the extraction method was assessed by determining whether it was possible to create a library that could be applied as a suitable template for a next-generation sequencer. In conclusion, a method was developed for consistent and accurate nucleic acid extraction from high-viscosity floricultural samples. These results demonstrate improved techniques for DNA and RNA extraction from flowers, help facilitate gene editing of floricultural species and expand the boundaries of research and commercial opportunities.

Keywords: floriculture, gene editing, next-generation sequencing, nucleic acid extraction

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379 Targeting Glucocorticoid Receptor Eliminate Dormant Chemoresistant Cancer Stem Cells in Glioblastoma

Authors: Aoxue Yang, Weili Tian, Yonghe Wu, Haikun Liu

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Brain tumor stem cells (BTSCs) are resistant to therapy and give rise to recurrent tumors. These rare and elusive cells are likely to disseminate during cancer progression, and some may enter dormancy, remaining viable but not increasing. The identification of dormant BTSCs is thus necessary to design effective therapies for glioblastoma (GBM) patients. Little progress has been made in therapeutic treatment of glioblastoma in the last decade despite rapid progress in molecular understanding of brain tumors1. Here we show that the stress hormone glucocorticoid is essential for the maintenance of brain tumor stem cells (BTSCs), which are resistant to conventional therapy. The glucocorticoid receptor (GR) regulates metabolic plasticity and chemoresistance of the dormant BTSC via controlling expression of GPD1 (glycerol-3-phosphate dehydrogenase 1), which is an essential regulator of lipid metabolism in BTSCs. Genomic, lipidomic and cellular analysis confirm that GR/GPD1 regulation is essential for BTSCs metabolic plasticity and survival. We further demonstrate that the GR agonist dexamethasone (DEXA), which is commonly used to control edema in glioblastoma, abolishes the effect of chemotherapy drug temozolomide (TMZ) by upregulating GPD1 and thus promoting tumor cell dormancy in vivo, this provides a mechanistic explanation and thus settle the long-standing debate of usage of steroid in brain tumor patient edema control. Pharmacological inhibition of GR/GPD1 pathway disrupts metabolic plasticity of BTSCs and prolong animal survival, which is superior to standard chemotherapy. Patient case study shows that GR antagonist mifepristone blocks tumor progression and leads to symptomatic improvement. This study identifies an important mechanism regulating cancer stem cell dormancy and provides a new opportunity for glioblastoma treatment.

Keywords: cancer stem cell, dormancy, glioblastoma, glycerol-3-phosphate dehydrogenase 1, glucocorticoid receptor, dexamethasone, RNA-sequencing, phosphoglycerides.

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378 Research Trends in Using Virtual Reality for the Analysis and Treatment of Lower-Limb Musculoskeletal Injury of Athletes: A Literature Review

Authors: Hannah K. M. Tang, Muhammad Ateeq, Mark J. Lake, Badr Abdullah, Frederic A. Bezombes

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There is little research applying virtual reality (VR) to the treatment of musculoskeletal injury in athletes. This is despite their prevalence, and the implications for physical and psychological health. Nevertheless, developments of wireless VR headsets better facilitate dynamic movement in VR environments (VREs), and more research is expected in this emerging field. This systematic review identified publications that used VR interventions for the analysis or treatment of lower-limb musculoskeletal injury of athletes. It established a search protocol, and through narrative discussion, identified existing trends. Database searches encompassed four term sets: 1) VR systems; 2) musculoskeletal injuries; 3) sporting population; 4) movement outcome analysis. Overall, a total of 126 publications were identified through database searching, and twelve were included in the final analysis and discussion. Many of the studies were pilot and proof of concept work. Seven of the twelve publications were observational studies. However, this may provide preliminary data from which clinical trials will branch. If specified, the focus of the literature was very narrow, with very similar population demographics and injuries. The trends in the literature findings emphasised the role of VR and attentional focus, the strategic manipulation of movement outcomes, and the transfer of skill to the real-world. Causal inferences may have been undermined by flaws, as most studies were limited by the practicality of conducting a two-factor clinical-VR-based study. In conclusion, by assessing the exploratory studies, and combining this with the use of numerous developments, techniques, and tools, a novel application could be established to utilise VR with dynamic movement, for the effective treatment of specific musculoskeletal injuries of athletes.

Keywords: athletes, lower-limb musculoskeletal injury, rehabilitation, return-to-sport, virtual reality

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377 An Affordability Evaluation of Computer-Based Social-Emotional Skills Interventions for School-Aged Children with Autism Spectrum Disorder

Authors: Ezra N. S. Lockhart

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The number of children diagnosed with autism spectrum disorder (ASD) has increased approximately 173% during the last decade making ASD the fastest growing developmental disability in the United States. This rise in prevalence rates indeed has an effect on schools. ASD is overwhelmingly the most reported primary special education eligibility category for students accessing special education, at a national average of 61.3%. ASD is regarded as an urgent public health concern at an estimated annual per capita cost of $3.2 million. Furthermore, considering that ASD is a lifelong disorder estimated lifetime per capita cost reach $35 billion. The resources available to special education programs are insufficient to meet the educational needs of the 6.4 million students receiving special educational services. This is especially true given that there has been and continues to be a chronic shortage of fully certified special education teachers for decades. Reports indicate that 81.1% of students with special needs spend 40% or more in general education classrooms. Regardless of whether support is implemented in the special education or general education classroom the resource demand is obvious. Schools are actively seeking to implement low-cost alternatives and budget saving measures in response to this demand. In public school settings, programs such as Applied Behavior Analysis are challenging to implement and fund at $40,000 per student per year. As an alternative, computer-based interventions are inexpensive, less time-consuming to implement, and require minimal teacher or paraprofessional training to administer. Affordability, pricing schemes, availability, and compatibility of computer-based interventions that support social and emotional skill development in individuals with ASD are discussed.

Keywords: affordability, autism spectrum disorder, computer-based intervention, emotional skills, social skills

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376 "Empowering Minds and Unleashing Curiosity: DIY Biotechnology for High School Students in the Age of Distance Learning"

Authors: Victor Hugo Sanchez Rodriguez

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Amidst the challenges posed by pandemic-induced lockdowns, traditional educational models have been disrupted. To bridge the distance learning gap, our project introduces an innovative initiative focused on teaching high school students basic biotechnology techniques. We aim to empower young minds and foster curiosity by encouraging students to create their own DIY biotechnology laboratories using easily accessible materials found at home. This abstract outlines the key aspects of our project, highlighting its importance, methodology, and evaluation approach.In response to the pandemic's limitations, our project targets the delivery of biotechnology education at a distance. By engaging students in hands-on experiments, we seek to provide an enriching learning experience despite the constraints of remote learning. The DIY approach allows students to explore scientific concepts in a practical and enjoyable manner, nurturing their interest in biotechnology and molecular biology. Originally designed to assess professional-level research programs, we have adapted the URSSA to suit the context of biotechnology and molecular biology synthesis for high school students. By applying this tool before and after the experimental sessions, we aim to gauge the program's impact on students' learning experiences and skill development. Our project's significance lies not only in its novel approach to teaching biotechnology but also in its adaptability to the current global crisis. By providing students with a stimulating and interactive learning environment, we hope to inspire educators and institutions to embrace creative solutions during challenging times. Moreover, the insights gained from our evaluation will inform future efforts to enhance distance learning programs and promote accessible science education.

Keywords: DIY biotechnology, high school students, distance learning, pandemic education, undergraduate research student self-assessment (URSSA)

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375 MicroRNA in Bovine Corpus Luteum during Early Pregnancy

Authors: Rreze Gecaj, Corina Schanzenbach, Benedikt Kirchner, Michael Pfaffl, Bajram Berisha

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The maintenance of corpus lutem (CL) during early pregnancy in cattle is a critical and multifarious process. A luteotrophic mechanism originating from the embryo is widely accepted as the triggering signal for the CL maintenance. In the cattle, it is the interferon-tau (IFNT) secretion form conceptus that prevents CL regression and ensures progesterone production for the establishment of pregnancy. In addition to endocrine and paracrine signals, microRNA (miRNA) can also support CL sustainability during early pregnancy. MiRNA are small non-coding nucleic acids that regulate gene expression post-transcriptionally and are shown to be involved in the modulation of CL function. However, the examination of miRNAs in corpus luteum function at the early pregnancy still remains largely uncovered. This study aims at profiling the expression of miRNA in CL during the early pregnancy in cattle by comparing it with the CL form late cycle and with the regressed CL. Corpora lutea were assigned in two different groups during the cycle (C13 group, late CL: days 13-18 and C18, regressed CL group: day >18) and during the early pregnancy (group P: 1-2 month). The estrous cycle was determined by macroscopic examination and to age the fetus crown-rump length measurement was applied. A total of 9 corpora lutea from individual animals were included in the study, three corpora lutea for each group. MiRNAs population was profiled using small RNA next-generation sequencing and biologically significant miRNAs were evaluated for their differential expression using the DESeq2-methodology. We show that 6 differentially expressed miRNAs (bta-mir-2890, -2332, -2441-3p, -148b, -1248 and -29c) are common to both comparisons, P vs C13 and P vs C18. While for each stage individually we have identified unique miRNAs differentially expressed only for the given comparison. bta-miR-23a and -769 were unique miRNAs differentially expressed in P vs C13, whereas forty-four unique miRNAs were identified as differentially expressed in P vs C18. These data confirm that miRNAs are highly abundant in luteal tissue during early pregnancy and potentially regulate the CL maintenance at this stage of fetus development.

Keywords: bovine, corpus luteum, microRNA, pregnancy, RNA-Seq

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374 An Analysis of Twitter Use of Slow Food Movement in the Context of Online Activism

Authors: Kubra Sultan Yuzuncuyil, Aytekin İsman, Berkay Bulus

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With the developments of information and communication technologies, the forms of molding public opinion have changed. In the presence of Internet, the notion of activism has been endowed with digital codes. Activists have engaged the use of Internet into their campaigns and the process of creating collective identity. Activist movements have been incorporating the relevance of new communication technologies for their goals and opposition. Creating and managing activism through Internet is called Online Activism. In this main, Slow Food Movement which was emerged within the philosophy of defending regional, fair and sustainable food has been engaging Internet into their activist campaign. This movement supports the idea that a new food system which allows strong connections between plate and planet is possible. In order to make their voices heard, it has utilized social networks and develop particular skills in the framework online activism. This study analyzes online activist skills of Slow Food Movement (SFM) develop and attempts to measure its effectiveness. To achieve this aim, it adopts the model proposed by Sivitandies and Shah and conduct both qualitiative and quantiative content analysis on social network use of Slow Food Movement. In this regard, the sample is chosen as the official profile and analyzed between in a three month period respectively March-May 2017. It was found that SFM develops particular techniques that appeal to the model of Sivitandies and Shah. The prominent skill in this regard was found as hyperlink abbreviation and use of multimedia elements. On the other hand, there are inadequacies in hashtag and interactivity use. The importance of this study is that it highlights and discusses how online activism can be engaged into a social movement. It also reveals current online activism skills of SFM and their effectiveness. Furthermore, it makes suggestions to enhance the related abilities and strengthen its voice on social networks.

Keywords: slow food movement, Twitter, internet, online activism

Procedia PDF Downloads 274
373 The Comparison Study of Human Microbiome in Chronic Rhinosinusitis between Adults and Children

Authors: Il Ho Park, Joong Seob Lee, Sung Hun Kang, Jae-Min Shin, Il Seok Park, Seok Min Hong, Seok Jin Hong

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Introduction: The human microbiota is the aggregate of microorganisms, and the bacterial microbiome of the human digestive tract contributes to both health and disease. In health, bacteria are key components in the development of mucosal barrier function and in innate and adaptive immune responses, and they also work to suppress the establishment of pathogens. In human upper airway, the sinonasal microbiota might play an important role in chronic rhinosinusitis (CRS). The purpose of this study is to investigate the human upper airway microbiome in CRS patients and to compare the sinonasal microbiome of adults with children. Materials and methods: A total of 19 samples from 19 patients (Group1; 9 CRS in children, aged 5 to 14 years versus Group 2; 10 CRS in adults aged 21 to 59 years) were examined. Swabs were collected from the middle meatus and/or anterior ethmoid region under general anesthesia during endoscopic sinus surgery or tonsillectomy. After DNA extraction from swab samples, we analysed bacterial microbiome consortia using 16s rRNA gene sequencing approach (the Illumina MiSeq platform). Results: In this study, relatively abundance of the six bacterial phyla and tremendous genus and species found in substantial amounts in the individual sinus swab samples, include Corynebacterium, Hemophilus, Moraxella, and Streptococcus species. Anaerobes like Fusobacterium and Bacteroides were abundantly present in the children group, Bacteroides and Propionibacterium were present in adults group. In genus, Haemophilus was the most common CRS microbiome in children and Corynebacterium was the most common CRS microbiome in adults. Conclusions: Our results show the diversity of human upper airway microbiome, and the findings will suggest that CRS is a polymicrobial infection. The Corynebacterium and Hemophilus may live as commensals on mucosal surfaces of sinus in the upper respiratory tract. The further study will be needed for analysis of microbiome-human interactions in upper airway and CRS.

Keywords: microbiome, upper airway, chronic rhinosinusitis, adult and children

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372 Characterisation of Extracellular Polymeric Substances from Bacteria Isolated from Acid Mine Decant in Gauteng, South Africa

Authors: Nonhlanhla Nkosi, Kulsum Kondiah

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The toxicological manifestation of heavy metals motivates interest towards the development of a reliable, eco-friendly biosorption process. With that being said, the aim of the current study was to characterise the EPS from heavy-metal resistant bacteria isolated from acid mine decant on the West Rand, Gauteng, South Africa. To achieve this, six exopolysaccharide (EPS) producing, metal resistant strains (Pb101, Pb102, Pb103, Pb204, Co101, and Ni101) were identified as Bacillus safensis strain NBRC 100820, Bacillus proteolyticus, Micrococcus luteus, Enterobacter sp. Pb204, Bacillus wiedmannii and Bacillus zhangzhouensis, respectively with 16S rRNA sequencing. Thereafter, EPS was extracted using chemical (formaldehyde/NaOH) and physical (ultrasonification) methods followed by physicochemical characterisation of carbohydrate, DNA, and protein contents using chemical assays and spectroscopy (FTIR- Fourier transformed infrared and 3DEEM- three-dimensional excitation-emission matrix fluorescence spectroscopy). EPS treated with formaldehyde/NaOH showed better recovery of macromolecules than ultrasonification. The results of the present study showed that carbohydrates were more abundant than proteins, with carbohydrate and protein concentrations of 8.00 mg/ml and 0.22 mg/ml using chemical method in contrast to 5.00 mg/ml and 0.77 mg/ml using physical method, respectively. The FTIR spectroscopy results revealed that the extracted EPS contained hydroxyl, amide, acyl, and carboxyl groups that corresponded to the aforementioned chemical analysis results, thus asserting the presence of carbohydrates, DNA, polysaccharides, and proteins in the EPS. These findings suggest that identified functional groups of EPS form surface charges, which serve as the binding sites for suspended particles, thus possibly mediating adsorption of divalent cations and heavy metals. Using the extracted EPS in the development of a cost-effective biosorption solution for industrial wastewater treatment is attainable.

Keywords: biosorbent, exopolysaccharides, heavy metals, wastewater treatment

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371 In vitro Modeling of Aniridia-Related Keratopathy by the Use of Crispr/Cas9 on Limbal Epithelial Cells and Rescue

Authors: Daniel Aberdam

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Haploinsufficiency of PAX6 in humans is the main cause of congenital aniridia, a rare eye disease characterized by reduced visual acuity. Patients have also progressive disorders including cataract, glaucoma and corneal abnormalities making their condition very challenging to manage. Aniridia-related keratopathy (ARK), caused by a combination of factors including limbal stem-cell deficiency, impaired healing response, abnormal differentiation, and infiltration of conjunctival cells onto the corneal surface, affects up to 95% of patients. It usually begins in the first decade of life resulting in recurrent corneal erosions, sub-epithelial fibrosis with corneal decompensation and opacification. Unfortunately, current treatment options for aniridia patients are currently limited. Although animal models partially recapitulate this disease, there is no in vitro cellular model of AKT needed for drug/therapeutic tools screening and validation. We used genome editing (CRISPR/Cas9 technology) to introduce a nonsense mutation found in patients into one allele of the PAX6 gene into limbal stem cells. Resulting mutated clones, expressing half of the amount of PAX6 protein and thus representative of haploinsufficiency were further characterized. Sequencing analysis showed that no off-target mutations were induced. The mutated cells displayed reduced cell proliferation and cell migration but enhanced cell adhesion. Known PAX6 targets expression was also reduced. Remarkably, addition of soluble recombinant PAX6 protein into the culture medium was sufficient to activate endogenous PAX6 gene and, as a consequence, rescue the phenotype. It strongly suggests that our in vitro model recapitulates well the epithelial defect and becomes a powerful tool to identify drugs that could rescue the corneal defect in patients. Furthermore, we demonstrate that the homeotic transcription factor Pax6 is able to be uptake naturally by recipient cells to function into the nucleus.

Keywords: Pax6, crispr/cas9, limbal stem cells, aniridia, gene therapy

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370 Self-Reliant Peer Learning for Nursing Students

Authors: U.-B. Schaer, M. Wehr, R. Hodler

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Background: Most nursing students require more training time for necessary nursing skills than defined by nursing schools curriculum to acquire basic nursing skills. Given skills training lessons are too brief to enable effective student learning, meaning in-depth skills practice and repetition various learning steps. This increases stress levels and the pressure to succeed for a nursing student with slower learning capabilities. Another possible consequence is that nursing students are less prepared in the required skills for future clinical practice. Intervention: The Bern College of Higher Education of Nursing, Switzerland, started the independent peer practice learning program in 2012. A concept was developed which defines specific aims and content as well as student’s rights and obligations. Students enlist beforehand and order the required materials. They organize themselves and train in small groups in allocated training location in the area of Learning Training and Transfer (LTT). During the peer practice, skills and knowledge can be repeatedly trained and reflected in the peer groups without the presence of a tutor. All invasive skills are practiced only on teaching dummies. This allows students to use all their potential. The students may access learning materials as literature and their own student notes. This allows nursing students to practice their skills and to deepen their knowledge on corresponding with their own learning rate. Results: Peer group discussions during the independent peer practice learning support the students in gaining certainty and confidence in their knowledge and skills. This may improve patient safety in future daily care practice. Descriptive statics show that the number of students who take advantage of the independent peer practice learning increased continuously (2012-2018). It has to be mentioned that in 2012, solely students of the first semester attended the independent peer practice learning program, while in 2018 over one-third of the participating students were in their fifth semester and final study year. It is clearly visible that the demand for independent peer practice learning is increasing. This has to be considered in the development of future teaching curricula.

Keywords: learning program, nursing students, peer learning, skill training

Procedia PDF Downloads 117
369 Genotypic Identification of Oral Bacteria Using 16S rRNA in Children with and without Early Childhood Caries in Kelantan, Malaysia

Authors: Zuliani Mahmood, Thirumulu Ponnuraj Kannan, Yean Yean Chan, Salahddin A. Al-Hudhairy

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Caries is the most common childhood disease which develops due to disturbances in the physiological equilibrium in the dental plaque resulting in demineralization of tooth structures. Plaque and dentine samples were collected from three different tooth surfaces representing caries progression (intact, over carious lesion and dentine) in children with early childhood caries (ECC, n=36). In caries free (CF) children, plaque samples were collected from sound tooth surfaces at baseline and after one year (n=12). The genomic DNA was extracted from all samples and subjected to 16S rRNA PCR amplification. The end products were cloned into pCR®2.1-TOPO® Vector. Five randomly selected positive clones collected from each surface were sent for sequencing. Identification of the bacterial clones was performed using BLAST against GenBank database. In the ECC group, the frequency of Lactobacillus sp. detected was significantly higher in the dentine surface (p = 0.031) than over the cavitated lesion. The highest frequency of bacteria detected in the intact surfaces was Fusobacterium nucleatum subsp. polymorphum (33.3%) while Streptococcus mutans was detected over the carious lesions and dentine surfaces at a frequency of 33.3% and 52.7% respectively. Fusobacterium nucleatum subsp. polymorphum was also found to be highest in the CF group (41.6%). Follow up at the end of one year showed that the frequency of Corynebacterium matruchotii detected was highest in those who remained caries free (16.6%), while Porphyromonas catoniae was highest in those who developed caries (25%). In conclusion, Streptococcus mutans and Porphyromonas catoniae are strongly associated with caries progression, while Lactobacillus sp. is restricted to deep carious lesions. Fusobacterium nucleatum subsp. polymorphum and Corynebacterium matruchotii may play a role in sustaining the healthy equilibrium in the dental plaque. These identified bacteria show promise as potential biomarkers in diagnosis which could help in the management of dental caries in children.

Keywords: early childhood caries, genotypic identification, oral bacteria, 16S rRNA

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368 Molecular Diversity of Forensically Relevant Insects from the Cadavers of Lahore

Authors: Sundus Mona, Atif Adnan, Babar Ali, Fareeha Arshad, Allah Rakha

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Molecular diversity is the variation in the abundance of species. Forensic entomology is a neglected field in Pakistan. Insects collected from the crime scene should be handled by forensic entomologists who are currently virtually non-existent in Pakistan. Correct identification of insect specimen along with knowledge of their biodiversity can aid in solving many problems related to complicated forensic cases. Inadequate morphological identification and insufficient thermal biological studies limit the entomological utility in Forensic Medicine. Recently molecular identification of entomological evidence has gained attention globally. DNA barcoding is the latest and established method for species identification. Only proper identification can provide a precise estimation of postmortem intervals. Arthropods are known to be the first tourists scavenging on decomposing dead matter. The objective of the proposed study was to identify species by molecular techniques and analyze their phylogenetic importance with barcoded necrophagous insect species of early succession on human cadavers. Based upon this identification, the study outcomes will be the utilization of established DNA bar codes to identify carrion feeding insect species for concordant estimation of post mortem interval. A molecular identification method involving sequencing of a 658bp ‘barcode’ fragment of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene from collected specimens of unknown dipteral species from cadavers of Lahore was evaluated. Nucleotide sequence divergences were calculated using MEGA 7 and Arlequin, and a neighbor-joining phylogenetic tree was generated. Three species were identified, Chrysomya megacephala, Chrysomya saffranea, and Chrysomya rufifacies with low genetic diversity. The fixation index was 0.83992 that suggests a need for further studies to identify and classify forensically relevant insects in Pakistan. There is an exigency demand for further research especially when immature forms of arthropods are recovered from the crime scene.

Keywords: molecular diversity, DNA barcoding, species identification, forensically relevant

Procedia PDF Downloads 142
367 Changes in the fecal Microbiome of Periparturient Dairy Cattle and Associations with the Onset of Salmonella Shedding

Authors: Lohendy Munoz-Vargas, Stephen O. Opiyo, Rose Digianantonio, Michele L. Williams, Asela Wijeratne, Gregory Habing

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Non-typhoidal Salmonella enterica is a zoonotic pathogen with critical importance in animal and public health. The persistence of Salmonella on farms affects animal productivity and health, and represents a risk for food safety. The intestinal microbiota plays a fundamental role in the colonization and invasion of this ubiquitous microorganism. To overcome the colonization resistance imparted by the gut microbiome, Salmonella uses invasion strategies and the host inflammatory response to survive, proliferate, and establish infections with diverse clinical manifestations. Cattle serve as reservoirs of Salmonella, and periparturient cows have high prevalence of Salmonella shedding; however, to author`s best knowledge, little is known about the association between the gut microbiome and the onset of Salmonella shedding during the periparturient period. Thus, the objective of this study was to assess the association between changes in bacterial communities and the onset of Salmonella shedding in cattle approaching parturition. In a prospective cohort study, fecal samples from 98 dairy cows originating from four different farms were collected at four time points relative to calving (-3 wks, -1 wk, +1 wk, +3 wks). All 392 samples were cultured for Salmonella. Sequencing of the V4 region of the 16S rRNA gene using the Illumina platform was completed to evaluate the fecal microbiome in a selected sample subset. Analyses of microbial composition, diversity, and structure were performed according to time points, farm, and Salmonella onset status. Individual cow fecal microbiomes, predominated by Bacteroidetes, Firmicutes, Spirochaetes, and Proteobacteria phyla, significantly changed before and after parturition. Microbial communities from different farms were distinguishable based on multivariate analysis. Although there were significant differences in some bacterial taxa between Salmonella positive and negative samples, our results did not identify differences in the fecal microbial diversity or structure for cows with and without the onset of Salmonella shedding. These data suggest that determinants other than the significant changes in the fecal microbiome influence the periparturient onset of Salmonella shedding in dairy cattle.

Keywords: dairy cattle, microbiome, periparturient, Salmonella

Procedia PDF Downloads 168