Search results for: early stages

Authors: Jeannette Anniés, Panagiotis Michalis, Chrysoula Papathanasiou, Selby Knudsen

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RiskPACC project aims to bring together researchers, practitioners, and first responders from nine European countries following a co-creation approach aiming to develop customised solutions to meet the needs of end-users. The co-creation workshops target to enhance the communication pathways between local civil protection authorities (CPAs) and citizens, in an effort to close the risk perception-action gap (RPAG). The participants in the workshops include a variety of stakeholders, as well as citizens, fostering the dialogue between the groups and supporting citizen participation in disaster risk management (DRM). The co-creation methodology in place implements co-design elements due to the integration of four ICT tools. Such ICT tools include web-based and mobile application technical solutions in different development stages, ranging from formulation and validation of concepts to pilot demonstrations. In total, seven different case studies are foreseen in RiskPACC. The workflow of the workshops is designed to be adaptive to every of the seven case study countries and their cultures’ particular needs. This work aims to provide an overview of the the preparation and the conduction of the workshops in which researchers and practitioners focused on mapping these different needs from the end users. The latter included first responders but also volunteers and citizens who actively participated in the co-creation workshops. The strategies to improve communication between CPAs and citizens themselves differ in the countries, and the modules of the co-creation methodology are adapted in response to such differences. Moreover, the project partners experienced how the structure of such workshops is perceived differently in the seven case studies. Therefore, the co-creation methodology itself is a design method underlying several iterations, which are eventually shaped by cross-cultural collaboration. For example, some case studies applied other modules according to the participatory group recruited. The participants were technical experts, teachers, citizens, first responders, or volunteers, among others. This work aspires to present the divergent approaches of the seven case studies implementing the co-creation methodology proposed, in response to different perceptions of the modules. An analysis of the adaptations and implications will also be provided to assess where the case studies’ objective of improving disaster resilience has been obtained.

Keywords: citizen participation, co-creation, disaster resilience, risk perception, ICT tools

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Authors: Kayondoamunmose Kamafrika

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Experimental based Engineering education approach plays a vital role in the development of student’s deep understanding of both social and physical sciences. Experimental based education approach through laboratory class activities prepare students to meet national demand for high-tech skilled individuals in the government and private sector. However, students across the country are faced with difficulties in classes that involve laboratory activities: poor experimental based exposure in their early development of student’s education-life-cycle, lack of student engagement in scientific method practical thinking approach, lack of communication between students and the instructor during class, a large number of students in one classroom, lack of instruments and improper equipment calibration. The purpose of this paper is to help students develop their own scientific knowledge and understanding, develop their methodologies in the design of experiments, collect and analyze data, write laboratory reports, present and explain their findings. Experimental based laboratory activities allow students to learn with high-level understanding as well as engage in the design processes of constructing knowledge through practical means of doing science. Experimental based education systems approach will act as a catalyst in the development of practical-based-educational methodologies in social and physical science and engineering domain of learning; thereby, converting laboratory classes into pilot industries and students into professional experts in finding a solution for complex problems, research, and development of super high- tech systems.

Keywords: experimental, engineering, innovation, practicability

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Authors: Zdzisław Kobos, Robert Jędrys, Zbigniew Wochyński

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Motor activity and good health are sine qua non determinants of a proper practice of the profession, especially aviation. Therefore, candidates to the aviation are selected according their psychomotor ability by both specialist medical commissions. Moreover, they must past an examination of the physical fitness. During the studies in the air force academy, eye-hand coordination is formed in two stages. The future aircraft pilots besides all-purpose physical education must practice specialist training on SAGI. Training includes: looping, aerowheel, and gyroscope. Aim of the training on the above listed apparatuses is to form eye-hand coordination during the tasks in the air. Such coordination is necessary to perform various figures in the real flight. Therefore, during the education of the future pilots, determinants of the effective ways of this important parameter of the human body functioning are sought for. Several studies of the sport psychology indicate an important role of the temperament as a factor determining human behavior during the task performance and acquiring operating skills> Polish psychologist Jan Strelau refers to the basic, relatively constant personality features which manifest themselves in the formal characteristics of the human behavior. Temperament, being initially determined by the inborn physiological mechanisms, changes in the course of maturation and some environmental factors and concentrates on the energetic level and reaction characteristics in time. Objectives. This study aimed at seeking a relationship between temperamental features and eye-hand coordination formation during training on SAGI. Material and Methods: Group of 30 students of pilotage was examined in two situations. The first assessment of the eye-hand coordination level was carried out before the beginning of a 30-hour training on SAGI. The second assessment was carried out after training completion. Training lasted for 2 hours once a week. Temperament was evaluated with The Formal Characteristics of Behavior − Temperament Inventory (FCB-TI) developed by Bogdan Zawadzki and Jan Strelau. Eye-hand coordination was assessed with a computer version of the Warsaw System of Psychological Tests. Results: It was found that the training on SAGI increased the level of eye-hand coordination in the examined students. Conclusions: Higher level of the eye-hand coordination was obtained after completion of the training. Moreover, a relationship between eye-hand coordination level and selected temperamental features was statistically significant.

Keywords: temperament, eye-hand coordination, pilot, SAGI

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Authors: Jasmal A. Syamsu, Muhammad Yusuf, Hikmah M. Ali, Mawardi A. Asja, Zulkharnaim

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This study was conducted and aimed in identifying and mapping the pasture as an ecological base of beef cattle. A survey was carried out during a period of April to June 2016, in Suppa, Mattirobulu, the district of Pinrang, South Sulawesi province. The mapping process of grazing area was conducted in several stages; inputting and tracking of data points into Google Earth Pro (version 7.1.4.1529), affirmation and confirmation of tracking line visualized by satellite with a variety of records at the point, a certain point and tracking input data into ArcMap Application (ArcGIS version 10.1), data processing DEM/SRTM (S04E119) with respect to the location of the grazing areas, creation of a contour map (a distance of 5 m) and mapping tilt (slope) of land and land cover map-making. Analysis of land cover, particularly the state of the vegetation was done through the identification procedure NDVI (Normalized Differences Vegetation Index). This procedure was performed by making use of the Landsat-8. The results showed that the topography of the grazing areas of hills and some sloping surfaces and flat with elevation vary from 74 to 145 above sea level (asl), while the requirements for growing superior grass and legume is an altitude of up to 143-159 asl. Slope varied between 0 - > 40% and was dominated by a slope of 0-15%, according to the slope/topography pasture maximum of 15%. The range of NDVI values for pasture image analysis results was between 0.1 and 0.27. Characteristics of vegetation cover of pasture land in the category of vegetation density were low, 70% of the land was the land for cattle grazing, while the remaining approximately 30% was a grove and forest included plant water where the place for shelter of the cattle during the heat and drinking water supply. There are seven types of graminae and 5 types of legume that was dominant in the region. Proportionally, graminae class dominated up 75.6% and legume crops up to 22.1% and the remaining 2.3% was another plant trees that grow in the region. The dominant weed species in the region were Cromolaenaodorata and Lantana camara, besides that there were 6 types of floor plant that did not include as forage fodder.

Keywords: pastoral, ecology, mapping, beef cattle

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Authors: Sumeeta Khurana, Kirti Megha, Amit Gupta, Rakesh Sehgal

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Background: Amoebic keratitis is a painful vision threatening infection caused by a free living pathogenic amoeba Acanthamoeba. It can be misdiagnosed and very difficult to treat if not suspected early. The epidemiology of Acanthamoeba genotypes causing infection in our geographical area is not yet known to the best of our knowledge. Objective: To characterize Acanthamoeba isolates from amoebic keratitis patients. Methods: A total of 19 isolates obtained from patients with amoebic keratitis presenting to the Advanced Eye Centre at Postgraduate Institute of Medical Education and Research, a tertiary care centre of North India over a period of last 10 years were included. Their corneal scrapings, lens solution and lens case (in case of lens wearer) were collected for microscopic examination, culture and molecular diagnosis. All the isolates were maintained in the Non Nutrient agar culture medium overlaid with E.coli and 13 strains were axenised and maintained in modified Peptone Yeast Dextrose Agar. Identification of Acanthamoeba genotypes was based on amplification of diagnostic fragment 3 (DF3) region of the 18srRNA gene followed by sequencing. Nucleotide similarity search was performed by BLAST search of sequenced amplicons in GenBank database (http//www.ncbi.nlm.nih.gov/blast). Multiple Sequence alignments were determined by using CLUSTAL X. Results: Nine out of 19 Acanthamoeba isolates were found to belong to Genotype T4 followed by 6 isolates of genotype T11, 3 T5 and 1 T3 genotype. Conclusion: T4 is the predominant Acanthamoeba genotype in our geographical area. Further studies should focus on differences in pathogenicity of these genotypes and their clinical significance.

Keywords: Acanthamoeba, free living amoeba, keratitis, genotype, ocular

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Authors: Hung Lin-Zin, Lai Mei-Yen

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Prostate cancer is the most commonly diagnosed male cancer in the U.S. The prevalence is around 1 in 8. The etiology of prostate cancer is still unknown, but some predisposing factors, such as age, black race, family history, and obesity, may increase the risk of the disease. In 2020, a total of 7,178 Taiwanese people were nearly diagnosed with prostate cancer, accounting for 5.88% of all cancer cases, and the incidence rate ranked fifth among men. In that year, the total number of deaths from prostate cancer was 1,730, accounting for 3.45% of all cancer deaths, and the death rate ranked 6th among men, accounting for 94.34% of the cases of male reproductive organs. Looking for domestic and foreign literature on the use of OMOP (Observational Medical Outcomes Partnership, hereinafter referred to as OMOP) database analysis, there are currently nearly a hundred literature published related to nursing-related health problems and nursing measures built in the OMOP general data model database of medical institutions are extremely rare. The OMOP common data model construction analysis platform is a system developed by the FDA in 2007, using a common data model (common data model, CDM) to analyze and monitor healthcare data. It is important to build up relevant nursing information from the OMOP- CDM database to assist our daily practice. Therefore, we choose prostate cancer patients who are our popular care objects and use the OMOP- CDM database to explore the common associated health problems. With the assistance of OMOP-CDM database analysis, we can expect early diagnosis and prevention of prostate cancer patients' comorbidities to improve patient care.

Keywords: OMOP, nursing diagnosis, health problem, prostate cancer

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Authors: Camille Intson

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This paper endeavors to explore the opportunities, challenges, and ethics of reconstructing and re-enacting archives of memory through virtual reality (VR) performance, using Jordan Tannahill’s Draw Me Close as an exemplary case study. Draw Me Close is a 1:1 virtual reality (VR) performance in which the artist’s childhood memories, experiences, and interactions with his mother are reconstructed in the wake of her passing. Solo audience members are positioned as Jordan (the subject and character) and taken through a series of narratives, (virtual) spaces, and interactions with his “mother,” played by a live actor. Piece by piece, audiences are brought into the world of the “shifting” archive, inhabiting Jordan’s reconstructed virtual world from his early explorations of queer sexuality through to his mother’s cancer diagnosis and passing. This paper will explore how the world of Draw Me Close represents a “touching” and/or “queering” of time within its archive, blurring and transgressing the boundaries between the animate and the inanimate, life and death. On a philosophical level, considering foundational queer performance scholarship and archival theory, it will also examine how performance’s ephemerality rewards its artists with the dual advantages of visibility and protection, allowing for an ethical exploration of traumatic memory and loss within a disappearing medium. Finally, this provocation will use Draw Me Close as a point of departure from which to outline future possibilities for performance and emerging technologies’ engagements with archival theory and practice. By positioning virtual reality (VR) as an archive-constructing medium, it aims to move beyond the question of how we can take performances seriously as archives towards how personal archive construction is itself a performative act.

Keywords: intermedial theatre, new media arts, queer performance, virtual reality

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Authors: Dipendra Kumar Yadav, Rajani Ghimire, Saroj Yadav

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Background: The concern about adolescent on reproductive health has grown due to unprecedented increasing rates of early pregnancies and sexually transmitted Infections and they do not have adequate awareness and knowledge about it. Access to these services as well as information about them is, therefore, crucial for adolescents to utilize and benefit from sexual and reproductive health services. The objective of the study was to assess the reproductive health knowledge, attitude and health services utilization among adolescents in rural and urban areas of Kaski district. Materials and Methods: A community-based descriptive cross-sectional study was conducted among adolescents (10-19 years of age) in rural and urban areas of Kaski district, Nepal. The period of data collection was October to November, 2014. Altogether 419 participants were taken for the study. Results: The mean age of the respondents was 15.86 and standard deviation was ±2.305. More than half (58.7 %) of the respondents were females and 41.3 % were males. Out of 419, majority (78.8%) of the respondents were known about family planning, among them only 70 % of respondents were aware about family planning methods. Fifty-one percentages of the respondents were aware about the sexually transmitted diseases. Before giving a birth there is need to consult with partner with this fact 68.7 % of the respondents were agree, 23.6 % of them were neutral and very few (7.6%) of them were disagree. Nearly twenty six percentage of the respondents were faced the reproductive health problems within one month. Out of 107 respondents, 57.9 % did not utilize reproductive health services because of different reasons. Conclusions: The overall level of knowledge towards reproductive health among adolescents was found low. However, levels of attitude towards different reproductive health components were found favorable. Only 42.1% of the respondents were utilized reproductive health services among those who was faced the reproductive health problems within one month which was low coverage of reproductive health services utilization.

Keywords: reproductive health knowledge, reproductive health attitudes, adolescent, service utilization

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Authors: C. Chaillou

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The dominant theoretical approaches in Criminology are interested in the phenomenon of delinquency from the question of the management of the risks incurred by the population. Thus, this research advocate prevention of this phenomenon by a tracking of early disorders in children. Treatments offered to rely on medical research (genetics and biology are cited as a reference) and assuming a high naturalization of delinquent behaviour. Programs that are offered also reduce to a recovery of the deviant behaviour, and rely readily on behavioral guidelines, with an educational grant. Public policy then rely on these programs to prevent unwanted behaviour within a given population and to reduce the risk for the company. This is the case in France, with national institutes making (juvenile) violence a public health problem. We consider that other approaches, issues of sociology, are more relevant to the treatment of offenders. These approaches are moving, not on its prevention, but from its inputs and its outputs. Several modalities of entries and exits of delinquency can find and analyze in terms of process. We assume that there is a dynamic inherent in the individual and it is important to take into account the environment of the offender. These different types of processes can illuminate from the derived work of the Psychoanalytical psychopathology and lead to more effective treatment of delinquent acts. Psychoanalytic concepts have enabled us to offer a new look means to treat delinquency, placing several types of relationship with the other and relating to the clinical structure and the uniqueness of the case, we have been able to enter subjective and unconscious logics at work in delinquent acts. This research has facilitated the reduction of these types of subjective responses and proposed others, opening to a reintegration of offenders in a social link them being more favourable and in a longer term.

Keywords: delinquency, insertion, social link, unconscious

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Authors: V. Reyes, P. Ferreira

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In response to high levels of competitiveness, industrial systems have evolved to improve productivity. As a consequence, a rapid increase in volume production and simultaneously, a customization process require lower costs, more variety, and accurate quality of products. Reducing time-cycle production, enabling customizability, and ensure continuous quality improvement are key features in advance intelligent industry. In this scenario, customers and producers will be able to participate in the ongoing production life cycle through real-time interaction. To achieve this vision, transparency, predictability, and adaptability are key features that provide the industrial systems the capability to adapt to customer demands modifying the manufacturing process through an autonomous response and acting preventively to avoid errors. The industrial system incorporates a diversified number of components that in advanced industry are expected to be decentralized, end to end communicating, and with the capability to make own decisions through feedback. The evolving process towards advanced intelligent industry defines a set of stages to empower components of intelligence and enhancing efficiency to achieve the decision-making stage. The integrated system follows an industrial cyber-physical system (CPS) architecture whose real-time integration, based on a set of enabler technologies, links the physical and virtual world generating the digital twin (DT). This instance allows incorporating sensor data from real to virtual world and the required transparency for real-time monitoring and control, contributing to address important features of the advanced intelligent industry and simultaneously improve sustainability. Assuming the industrial CPS as the core technology toward the latest advanced intelligent industry stage, this paper reviews and highlights the correlation and contributions of the enabler technologies for the operationalization of each stage in the path toward advanced intelligent industry. From this research, a real-time integration architecture for a cyber-physical system with applications to collaborative robotics is proposed. The required functionalities and issues to endow the industrial system of adaptability are identified.

Keywords: cyber-physical systems, digital twin, sensor data, system integration, virtual model

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Authors: Ervin Meqikukiqi

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It is difficult to assess how athletes can best modify their movements to prevent non contact ACL injuries. Speaking with an athletic trainer, physical therapist, or sports medicine specialist is a good place to start. Recent research has allowed therapists and clinicians to easily identify and target weak muscle areas (e.g., weak hips, which leads to knock-kneed landing positions) and identify ways to improve strength and thus help prevent injury. In addition, other risk factors such as reduced hamstring strength and increased joint range of motion can be further assessed by a physical therapist or athletic trainer to improve performance-or rehabilitation efforts after an injury has occurred. Current studies also demonstrate that specific types of training, such as jump routines and learning to pivot properly, help athletes prevent ACL injuries. These types of exercises and training programs are more beneficial if athletes start when they are young. It may be optimal to integrate prevention programs during early adolescence, prior to when young athletes develop certain habits that increase the risk of an ACL injury. This is a 20 minute program designed to reduce the risk of tears of the Anterior Cruciate Ligament. It should be started at least four and preferably six weeks prior to start of competition.Ideally it is done five times per week preseason and three times per week in season.The coach or trainer must constantly observe athletes during these exercises to correct and maintain proper technique. Once the athletes understand the principles, they can monitor and coach each other. Four phases: Warm-up, Strengthening, Plyometrics, Agility and Balance.

Keywords: athletes, acl, prevention, injuries, plyoemtric, proprioception, agillity

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Authors: Lilian A. Okeke, I. Okeke, N. Waziri, S. Balogun, P. Nguku, O. Fawole

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Background: Access to ante-natal care services promotes safe motherhood and delivery with improved maternal and neonatal outcome. We conducted this study to identify factors influencing the utilization of antenatal care (ANC) and health delivery services. Methods: We conducted a cross sectional study. Households were numbered and a one in three sample was selected using a systematic sampling method. One hundred and ninety eight women who were either pregnant or had previous deliveries were interviewed using pretested structured questionnaires to obtain information on their socio-demographic characteristics, and reasons for non-utilization of ANC and health delivery services. We performed univariate and bivariate analysis using Epi info version 3.5.3. Results: The age of respondents ranged from (17-55 years) with a median age of 29 years. One hundred and ninety two (97%) utilized antenatal care services. Ninety three (47.9%) attended ANC at second trimester. More than half (58.6%) had ≥ 4 visits to ANC. One hundred and thirty one (66.2%) had their last delivery at home by a traditional birth attendant. Factors associated with ANC and health facility delivery services utilization were: age group 45-55 (OR 0.01; 95% CI: 0.00-0.16) and > 55 years (OR 0.03; 95% CI: 0.00-0.60), wife’s educational status (OR 3.17; 95% CI: 1.66-8.30), husband’s permission (OR 11.8; 95% CI 2.19-63.62), and distance ≥ 5km (OR 0.33; 95% CI: 0.16-0.60). Conclusion: ANC services were well utilized. Most women did not book early and had their last delivery at home. Predictors of ANC use and health facility delivery were age, wife’s educational status, husband's permission and long distance from health facility. A one-day health sensitization of the benefits of ANC utilization and the dangers of delivering at home was implemented.

Keywords: ante natal care, health facility, delivery services, rural area, Plateau state

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Authors: Theo Heyns Veldsman, Dieter Veldsman

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Organisational Identity (OI) refers to who and what the organisation is, what it stands for and does, and what it aspires to become. OI explores the perspectives of how we see ourselves, are seen by others and aspire to be seen. It provides as rationale the ‘why’ for the organisation’s continued existence. The most widely accepted differentiating features of OI are encapsulated in the organisation’s core, distinctive, differentiating, and enduring attributes. OI finds its concrete expression in the organisation’s Purpose, Vision, Strategy, Core Ideology, and Legacy. In the emerging new order infused by hyper-turbulence and hyper-fluidity, the VICCAS world, OI provides a secure anchor and steady reference point for the organisation, particularly the growing widespread focus on Purpose, which is indicative of the organisation’s sense of social citizenship. However, the transforming world of work (TWOW) - particularly the potent mix of ongoing disruptive innovation, the 4th Industrial Revolution, and the gig economy with the totally unpredicted COVID19 pandemic - has resulted in the consequential adoption of different workplace strategies by organisations in terms of how, where, and when work takes place. Different employment relations (transient to permanent); work locations (on-site to remote); work time arrangements (full-time at work to flexible work schedules); and technology enablement (face-to-face to virtual) now form the basis of the employer/employee relationship. The different workplace strategies, fueled by the demands of TWOW, pose a substantive challenge to organisations of doing durable OI work, able to fulfill OI’s critical attributes of core, distinctive, differentiating, and enduring. OI work is contained in the ongoing, reciprocally interdependent stages of sense-breaking, sense-giving, internalisation, enactment, and affirmation. The objective of our paper is to explore how to do durable OI work relative to different workplace strategies in the TWOW. Using a conceptual-theoretical approach from a practice-based orientation, the paper addresses the following topics: distinguishes different workplace strategies based upon a time/place continuum; explicates stage-wise the differential organisational content and process consequences of these strategies for durable OI work; indicates the critical success factors of durable OI work under these differential conditions; recommends guidelines for OI work relative to TWOW; and points out ethical implications of all of the above.

Keywords: organisational identity, workplace strategies, new world of work, durable organisational identity work

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Authors: Preeti Nakhat

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Background: Values are essential part of one's life. They are inculcated since the early years of life and shape the personality of the individual. They play a tremendous role in decision making. Teenagers are seen perplexed about the values of their life. The challenge faced by majority of the teenage population in choosing between a positive and negative value is high. The values they adopt remain throughout their life and in every decision, hence it is a crucial topic of research. Research Methodology: This research aimed at finding out the value conflict of teenagers in relations. Hypothesis of the study are: H₀- There is no significant association between the life values and value conflict of higher secondary students; H₁– There is a significant association between the life values and value conflict of higher secondary students. For the same, the standardized tool, value conflict scale by R. L. Bhardwaj has been used. The tool consists 24 questions of different life situations with multiple choice options. Findings: There is 96% variation in value conflict due to evasion vs. fortitude, dependence vs. self-reliance, selfishness vs. probity, hate vs. love, fear vs. assertion and pragmatism vs. idealism life values. There is a positive association between all the life values and value conflict of higher secondary school students. Percentages of association are: 0.17% between value conflict and evasion vs. fortitude value, 0.16% between value conflict and dependence vs. self-reliance value, 0.17% between value conflict and selfishness vs. probity value, 0.16% between value conflict and hate vs. love value, 0.17% between value conflict and fear vs. assertion, 0.17% between value conflict and pragmatism vs. idealism value. Discussions: The dilemma faced by the students regarding value conflict is high. Bewilderment of being honest or lying, of loving or hating family and friends, being pragmatic or idealistic in life decision, being selfish or selfless is seen among the students. It is the challenge for the future. Teaching of values with a practical aspect should be added in the school curriculum.

Keywords: dilemma, conflict, school, values

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Authors: Krzysztof J. Czarnocki, F. Silveira, E. Czarnocka, K. Szaniawska

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Research background: Problems related to the occupational health and decreasing level of safety occur commonly in the construction industry. Important factor in the occupational safety in construction industry is scaffold use. All scaffolds used in construction, renovation, and demolition shall be erected, dismantled and maintained in accordance with safety procedure. Increasing demand for new construction projects unfortunately still is linked to high level of occupational accidents. Therefore, it is crucial to implement concrete actions while dealing with scaffolds and risk assessment in construction industry, the way on doing assessment and liability of assessment is critical for both construction workers and regulatory framework. Unfortunately, professionals, who tend to rely heavily on their own experience and knowledge when taking decisions regarding risk assessment, may show lack of reliability in checking the results of decisions taken. Purpose of the article: The aim was to indicate crucial parameters that could be modeling with Risk Assessment Model (RAM) use for improving both building enterprise productivity and/or developing potential and safety climate. The developed RAM could be a benefit for predicting high-risk construction activities and thus preventing accidents occurred based on a set of historical accident data. Methodology/Methods: A RAM has been developed for assessing risk levels as various construction process stages with various work trades impacting different spheres of enterprise activity. This project includes research carried out by teams of researchers on over 60 construction sites in Poland and Portugal, under which over 450 individual research cycles were carried out. The conducted research trials included variable conditions of employee exposure to harmful physical and chemical factors, variable levels of stress of employees and differences in behaviors and habits of staff. Genetic modeling tool has been used for developing the RAM. Findings and value added: Common types of trades, accidents, and accident causes have been explored, in addition to suitable risk assessment methods and criteria. We have found that the initial worker stress level is more direct predictor for developing the unsafe chain leading to the accident rather than the workload, or concentration of harmful factors at the workplace or even training frequency and management involvement.

Keywords: safety climate, occupational health, civil engineering, productivity

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Authors: Vilma Zydziunaite, Simona Kersiene

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The concept of focused leadership means that the leader gathers the entire community in various ways to communicate and cooperate with each other, to share their knowledge and responsibility, to get involved in problem-solving, to create a safe and trusting environment and to satisfy the needs and interests of each community member. The study's aim is to analyze the teamwork of teachers working in kindergartens and schools and its CEOs by implementing confused leadership. A mixed research design was used for the research study. Quantitative research used the teamwork test "Team-Puls" (2003). Data is processed by the IBM SPSS version 29.0 software package. Semi-structured interviews were used for data collection, and qualitative content analysis was applied for data analysis. The results of quantitative research show that there is no statistically significant difference between the evaluation averages of kindergarten and school teachers. Likewise, the effectiveness and evaluation of teacher teamwork in educational institutions depend on different characteristics and processes, such as the number of participating teachers, the involvement of the institution's administration or the stages of team formation. In the qualitative research, the components of the focused leadership categories applied by the kindergarten and school CEOs emerged. The categories reflect the components of shared leadership. In the study, the sharing of responsibilities and cooperation among teachers and the sharing of knowledge among themselves is distinguished. This shows that the action takes place between the teachers when they participate in the processes voluntarily, according to their wishes or for certain reasons. Distributed leadership components occurs when leadership responsibility is extended beyond the school CEO. The components of servant leadership are expressed when the CEO achieves organizational goals in the service of others. Servant leadership is helping and striving for others, creating a safe environment. The level of the educational institution does not affect working teachers in the evaluation of working in a team. Giving freedom to teachers, the role of the CEO is dividing responsibilities and creating cooperation between teachers as well as ensuring teachers' interests, needs, emotional well-being and professional development.

Keywords: teamwork, school, teacher, school CEO, school environment, mixed research, Team-Puls test, semi-structured interview, questioning survey, qualitative content analysis, focused leadership, teacher leadership

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Authors: Eman G. Alayad, Mazen G. Aleyad, Mohammed Alshahrani, Ibrahim Alnaami

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Introduction: Cerebral venous thrombosis (CVT) is a rare type of cerebrovascular disease that can occur at any age. Patients with CVT commonly present with headache, focal neurological deficit, decreased level of consciousness and seizures. Many etiologic risk factors have been reported for CVT, high altitude and oral contraceptive pill some of them. Case Presentation: A 37-year-old woman living in Abha city in the southeastern area of Saudi Arabia. (about 10,000 feet-3000 m) over the sea. complaining acute onset of severe diffuse headache and generalized tonic clonic convulsions. Followed by loss of consciousness. She was on contraceptive pills for the last 3 years. No significant Medical or surgical history. Brain CT revealed subarachnoid hemorrhage, with MRI findings showing thrombosis in transvers sinus. There was no vascular malformations such as aneurysm, arteriovenous malformation (AVM), or dural arteriovenous fistula. A CVT with subarachnoid hemorrhage was our final diagnosis based on clinical presentation and radiographic findings. Discussion: Patients with CVT had evidence of cortical SAH by 10 of 233, others found 3% of SAH was caused by CVT, indicating that the presence of cortical SAH without involvement of the basal cisterns may provide an early sign of underlying CVT. However, what is more interesting in this case, is the relationship of high altitude with CVT and SAH, which previously undescribed. Conclusion: High-altitude climbing per se was described as a risk factor for the development of CVT, though its occurrence was probably rare. Whether it is primary in etiology due to high altitude induced hypercoagulable state of unknown origin or due to cerebrovascular disturbances there is a need for further investigation especially at this unusual presentation of subarachnoid hemorrhage.

Keywords: cerebral venous thrombosis, high-altitude, subarachnoid hemorrhage, stroke

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Authors: Naim Izet Kajtazi

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Context: Frequently, the cause of raised intracranial pressure remains unresolved and rarely is related to spinal tumors, moreover less to spinal medulloblastoma without primary brain focus. Process: An 18-year-old woman had a 3-month history of headaches and impaired vision. Neurological examination revealed bilateral sixth cranial nerve palsies with bilateral papilloedema of grade III. No focal brain or spine lesion was found on imaging. Consecutive lumbar punctures showed high opening pressure and subsequent increasing protein level. The meningeal biopsy was negative. At one point, she developed an increasing headache, vomiting and back pain. Spine MRI showed diffuse nodular leptomeningeal enhancement with the largest nodule at T6–T7. Malignant cells were detected in cerebrospinal fluid. She underwent laminectomy with excisional biopsy, and pathology showed medulloblastoma WHO grade IV. Outcome: She was treated with chemotherapy and craniospinal irradiation and made a good recovery. Relevance: Primary spinal leptomeningeal medulloblastoma is extremely rare, especially without primary brain focus, but may cause increased intracranial pressure, even in the early microscopic phases, and it should be considered in the differential diagnosis if conventional and aggressive treatment of idiopathic intracranial hypertension fails. We assume that arachnoiditis from tumor seeding caused increased intracranial pressure. Appropriate neurosurgical intervention and surgical biopsy are mandated if a suspicious lesion is detected. Consider proper rescreening of the whole neuroaxis in refractory cases of intracranial hypertension.

Keywords: CNS infection, IIH, headache, primary spinal leptomeningeal medulloblastoma

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Authors: Shahzad Bhatti, M. Aslamkhan, Sana Abbas, Marcella Attimonelli, Hikmet Hakan Aydin, Erica Martinha Silva de Souza,

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Despite being situated at the crossroad of Asia, Pakistan has gained crucial importance because of its pivotal role in subsequent migratory events. To highlight the genetic footprints and to contribute an enigmatic picture of the relative population expansion pattern among four major Pashtun tribes in Khyber Pakhtunkhwa viz., Bangash, Khattak, Mahsuds and Orakzai, the complete mitochondrial control region of 100 Pashtun were analyzed. All Pashtun tribes studied here revealed high genetic diversity; that was comparable to the other Central Asian, Southeast Asian and European populations. The configuration of genetic variation and heterogeneity further unveiled through Multidimensional Scaling, Principal Component Analysis, and phylogenetic analysis. The results revealed that the Pashtun is a composite mosaic of West Eurasian ancestry of numerous geographic origin. They received substantial gene flow during different invasions and have a high element of the Western provenance. The most common haplogroups reported in this study are: South Asian haplogroup M (28%) and R (8%); whereas, West Asians haplogroups are present, albeit in high frequencies (67%) and widespread over all; HV (15%), U (17%), H (9%), J (8%), K (8%), W (4%), N (3%) and T (3%). Herein we linked the unexplored genetic connection between Ashkenazi Jews and Pashtun. The presence of specific haplotypes J1b (4%) and K1a1b1a (5%) point to a genetic connection of Jewish conglomeration with Khattak tribe. This was a result of an ancient genetic influx in the early Neolithic period that led to the formation of a diverse genetic substratum in present day Pashtun.

Keywords: mtDNA haplogroups, control region, Pakistan, KPK, ethnicity

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Authors: Francimar Alves, Marcos Castro, Marllus Lustosa

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In the context of the teaching of computer programming, the choice of tool to use is very important in the initiation and continuity of learning a programming language. The literature tools do not always provide usability and pedagogical dynamism clearly and accurately for effective learning. This hypothesis implies fall in productivity and difficulty of learning a particular programming language by students. The integrated development environments (IDEs) Dev-C ++ and Code :: Blocks are widely used in introductory courses for undergraduate courses in Computer Science for learning C and C ++ languages. However, after several years of discontinuity maintaining the source code of Dev-C ++ tool, the continued use of the same in the teaching and learning process of the students of these institutions has led to difficulties, mainly due to the lack of update by the official developers, which resulted in a sequence of problems in using it on educational settings. Much of the users, dissatisfied with the IDE Dev-C ++, migrated to Code :: Blocks platform targeting the more dynamic range in the learning process of the C and C ++ languages. Nevertheless, there is still the need to create a tool that can provide the resources of most IDE's software development literature, however, more interactive, simple, accurate and efficient. This motivation led to the creation of Falcon C ++ tool, IDE that brings with features that turn it into an educational platform, which focuses primarily on increasing student learning index in the early disciplines of programming and algorithms that use the languages ​​C and C ++ . As a working methodology, a field research to prove the truth of the proposed tool was used. The test results and interviews with entry-level students and intermediate in a postsecondary institution gave basis for the composition of this work, demonstrating a positive impact on the use of the tool in teaching programming, showing that the use of Falcon C ++ software is beneficial in the teaching process of the C and C ++ programming languages.

Keywords: ide, education, learning, development, language

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Authors: Chu Wan-Loy, Kok Yih-Yih, Choong Siew-Ling

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The human health risks due to poor air quality caused by a wide array of microorganisms have attracted much interest. Airborne algae have been reported as early as 19th century and they can be found in the air of tropic and warm atmospheres. Airborne algae normally originate from water surfaces, soil, trees, buildings and rock surfaces. It is estimated that at least 2880 algal cells are inhaled per day by human. However, there are relatively little data published on airborne algae and its related adverse health effects except sporadic reports of algae associated clinical allergenicity. A collection of airborne algae cultures has been established following a recent survey on the occurrence of airborne algae in indoor and outdoor environments in Kuala Lumpur. The aim of this study was to investigate the allergenic potential of the isolated airborne green and blue-green algae, namely Scenedesmus sp., Cylindrospermum sp. and Hapalosiphon sp.. The suspensions of freeze-dried airborne algae were adminstered into balb-c mice model through intra-nasal route to determine their allergenic potential. Results showed that Scenedesmus sp. (1 mg/mL) increased the systemic Ig E levels in mice by 3-8 fold compared to pre-treatment. On the other hand, Cylindrospermum sp. and Hapalosiphon sp. at similar concentration caused the Ig E to increase by 2-4 fold. The potential of airborne algae causing Ig E mediated type 1 hypersensitivity was elucidated using other immunological markers such as cytokine interleukin (IL)- 4, 5, 6 and interferon-ɣ. When we compared the amount of interleukins in mouse serum between day 0 and day 53 (day of sacrifice), Hapalosiphon sp. (1mg/mL) increased the expression of IL4 and 6 by 8 fold while the Cylindrospermum sp. (1mg/mL) increased the expression of IL4 and IFɣ by 8 and 2 fold respectively. In conclusion, repeated exposure to the three selected airborne algae may stimulate the immune response and generate Ig E in a mouse model.

Keywords: airborne algae, respiratory, allergenic, immune response, Malaysia

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Authors: Davendra Singh Malik

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India is one of the mega-biodiversity countries in the world and contributing about 11.72% of global fish diversity. The Yamuna river is the longest tributary of Ganga river ecosystem, providing a natural habitat for existing fish diversity of Himalayan region of Indian subcontinent. The several hydropower dams and barrages have been constructed on different locations of major rivers in Garhwal region. These dams have caused a major ecological threat to change existing fresh water ecosystems altering water flows, interrupting ecological connectivity, fragmenting habitats and native riverine fish species. Mahseer fishes (Indian carp) of the genus Tor, are large cyprinids endemic to continental Asia popularly known as ‘Game or sport fishes’ have continued to be decimated by fragmented natural habitats due to damming the water flow in riverine system and categorized as threatened fishes of India. The fresh water fish diversity as 24 fish species were recorded from Yamuna river. The present fish catch data has revealed that mahseer fishes (Tor tor and Tor putitora) were contributed about 32.5 %, 25.6 % and 18.2 % in upper, middle and lower riverine stretches of Yaumna river. The length range of mahseer (360-450mm) recorded as dominant size of catch composition. The CPUE (catch per unit effort) of mahseer fishes also indicated about a sharp decline of fish biomass, changing growth pattern, sex ratio and maturity stages of fishes. Only 12.5 – 14.8 % mahseer female brooders have showed only maturity phases in breeding months. The fecundity of mature mahseer female fish brooders ranged from 2500-4500 no. of ova during breeding months. The present status of mahseer fishery has attributed to the over exploitative nature in Yamuna river. The mahseer population is shrinking continuously in down streams of Yamuna river due to cumulative effects of various ecological stress. Mahseer conservation programme have implemented as 'in situ fish conservation' for enhancement of viable population size of mahseer species and restore the genetic loss of mahseer fish germplasm in Yamuna river of Garhwal Himalayan region.

Keywords: conservation practice, population dynamics, tor fish species, Yamuna River

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Authors: Ange Weinrabe, James Tran, Ian B. Hickie

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Risk-taking behaviour is common during youth. In the time between adolescence and early adulthood, young people (aged 15-25 years) are more vulnerable to mood disorders, such as anxiety and depression. What impact does an emerging mood disorder have on decision-making in youth at critical decision points in their lives? In this article, we explore the impact of risk and ambiguity on youth decision-making in a clinical setting using a well-known economic experiment. At two time points, separated by six to eight weeks, we measured risky and ambiguous choices concurrently with findings from three psychological questionnaires, the 10-item Kessler Psychological Distress Scale (K10), the 17-item Quick Inventory of Depressive Symptomatology Adolescent Version (QIDS-A17), and the 12-item Somatic and Psychological Health Report (SPHERE-12), for young help seekers aged 16-25 (n=30, mean age 19.22 years, 19 males). When first arriving for care, we found that 50% (n=15) of participants experienced severe anxiety (K10 ≥ 30) and were severely depressed (QIDS-A17 ≥ 16). In Session 2, taking attrition rates into account (n=5), we found that 44% (n=11) remained severe across the full battery of questionnaires. When applying multiple regression analyses of the pooled sample of observations (N=55), across both sessions, we found that participants who rated severely anxious avoided making risky decisions. We suggest there is some statistically significant (although weak) (p=0.09) relation between risk and severe anxiety scores as measured by K10. Our findings may support working with novel tools with which to evaluate youth experiencing an emerging mood disorder and their cognitive capacities influencing decision-making.

Keywords: anxiety, decision-making, risk, adolescence

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Authors: Alana E. Jansen

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With such a large proportion of organisations relying on the use of team-based structures, it is surprising that so few teams would be classified as high-performance teams. While the impact of team composition on performance has been researched frequently, there have been conflicting findings as to the effects, particularly when examined alongside other team factors. To broaden the theoretical perspectives on this topic and potentially explain some of the inconsistencies in research findings left open by other various models of team effectiveness and high-performing teams, the present study aims to use the Job-Demands-Resources model, typically applied to burnout and engagement, as a framework to examine how team composition factors (particularly diversity in team member characteristics) can facilitate or hamper team effectiveness. This study used a virtual interview design where participants were asked to both rate and describe their experiences, in one high-performing and one low-performing team, over several factors relating to demands, resources, team composition, and team effectiveness. A semi-structured interview protocol was developed, which combined the use of the Likert style and exploratory questions. A semi-targeted sampling approach was used to invite participants ranging in age, gender, and ethnic appearance (common surface-level diversity characteristics) and those from different specialties, roles, educational and industry backgrounds (deep-level diversity characteristics). While the final stages of data analyses are still underway, thematic analysis using a grounded theory approach was conducted concurrently with data collection to identify the point of thematic saturation, resulting in 35 interviews being completed. Analyses examine differences in perceptions of demands and resources as they relate to perceived team diversity. Preliminary results suggest that high-performing and low-performing teams differ in perceptions of the type and range of both demands and resources. The current research is likely to offer contributions to both theory and practice. The preliminary findings suggest there is a range of demands and resources which vary between high and low-performing teams, factors which may play an important role in team effectiveness research going forward. Findings may assist in explaining some of the more complex interactions between factors experienced in the team environment, making further progress towards understanding the intricacies of why only some teams achieve high-performance status.

Keywords: diversity, high-performing teams, job demands and resources, team effectiveness

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Authors: Virginia Andrea Garrido Meirelles

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Noah Webster was a lexicographer and a language reformer and is considered the ‘Father of American Scholarship and Education’ because of the exceptional contributions he made as a teacher and grammarian. The goal of this study is to show that the success of his plan can be explained by the fact that it matched the language-in-education policies of his time. To accomplish that goal the present study analyzes the Massachusetts School Laws of 1642, 1647 and 1648 and compares them to the preface of the first edition of The Grammatical Institute of the English Language. The referred laws were three legislative acts enacted in the Massachusetts Colony and replicated almost identically in the other New England colonies. The purpose of those laws was to eradicate pauperism and poverty, on the one side, and to disseminate the idea of right citizenship, on the other. However, until the Declaration of Independence in 1776, all the primers used in the colony were printed in Britain. In 1783, Noah Webster published the first part of his Grammatical Institute of the English Language. In this book, the author states that his goal is to promote the republican principles that guide the civil rights of that time. The material included many texts taken from the Bible to inspire aversion to inadequate behavior and preference for service and good manners. In addition, its goal was to present ‘a new plan of reducing the pronunciation of our language to an easy standard,’ and in that way, create a unified language to abolish ignorance and language corruption. The comparison between the laws and Webster’s Spelling Book shows that the book is the result of the historical and political situation when it was conceived and it satisfied the requirements of the language-in-education policies of the time.

Keywords: American English, language policy, the Massachusetts school laws, webster's spelling book

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Authors: Ian Omung'a

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Breast cancer remains to be one of the deadliest cancers for women worldwide, with the risk of developing tumors being as high as 50 percent in Sub-Saharan African countries like Kenya. With as many as 42 percent of these cases set to be diagnosed late when cancer has metastasized and or the prognosis has become terminal, Full Field Digital [FFD] Mammography remains an effective screening technique that leads to early detection where in most cases, successful interventions can be made to control or eliminate the tumors altogether. FFD Mammograms have been proven to multiply more effective when used together with Computer-Aided Detection and Diagnosis [CADe] systems, relying on algorithmic implementations of Deep Learning techniques in Computer Vision to carry out deep pattern recognition that is comparable to the level of a human radiologist and decipher whether specific areas of interest in the mammogram scan image portray abnormalities if any and whether these abnormalities are indicative of a benign or malignant tumor. Within this paper, we review emergent Deep Learning techniques that will prove relevant to the development of State-of-The-Art FFD Mammogram CADe systems. These techniques will span self-supervised learning for context-encoded occlusion, self-supervised learning for pre-processing and labeling automation, as well as the creation of a standardized large-scale mammography dataset as a benchmark for CADe systems' evaluation. Finally, comparisons are drawn between existing practices that pre-date these techniques and how the development of CADe systems that incorporate them will be different.

Keywords: breast cancer diagnosis, computer aided detection and diagnosis, deep learning, whole mammogram classfication, ultrasound classification, computer vision

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Authors: Ashit Syngle, Inderjit Verma, Pawan Krishan

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Objective: Endothelial progenitor cells (EPCs) have reparative potential in overcoming the endothelial dysfunction and reducing cardiovascular risk. EPC depletion has been demonstrated in the setting of established atherosclerotic diseases. With this background, we evaluated whether reduced EPCs population are associated with endothelial dysfunction, subclinical atherosclerosis and inflammatory markers in ankylosing spondylitis (AS) patients without any known traditional cardiovascular risk factor in AS patients. Methods: Levels of circulating EPCs (CD34+/CD133+), brachial artery flow-mediated dilatation, carotid intima-media thickness (CIMT) and inflammatory markers i.e erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), tissue necrosis factor (TNF)–α, interleukin (IL)-6, IL-1 were assessed in 30 AS patients (mean age33.41 ± 10.25; 11 female and 19 male) who fulfilled the modified New York diagnostic criteria with 25 healthy volunteers (mean age 29.36± 8.64; 9 female and 16 male) matched for age and sex. Results: EPCs (CD34+/CD133+) cells were significantly (0.020 ± 0.001% versus 0.040 ± 0.010%, p<0.001) reduced in patients with AS compared to healthy controls. Endothelial function (7.35 ± 2.54 versus 10.27 ±1.73, p=0.002), CIMT (0.63 ± 0.01 versus 0.35 ± 0.02, p < 0.001) and inflammatory markers were also significantly (p < 0.01) altered as compared to healthy controls. Specifically, CD34+CD133+cells were inversely multivariate correlated with CRP and TNF-α and endothelial dysfunction was positively correlated with reduced number of EPC. Conclusion: Depletion of EPCs population is an independent predictor of endothelial dysfunction and early atherosclerosis in AS patients and may provide additional information beyond conventional risk factors and inflammatory markers.

Keywords: endothelial progenitor cells, atherosclerosis, ankylosing spondylitis, cardiovascular

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Authors: Ranitha Guna Selvi D, Ramakrishnan R, Mohideen Abdul Kader

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Purpose: To study the role of Vision centers in managing preventable blindness through refractive error correction in Rural South India. Methods: A retrospective analysis of patients attending 15 Vision centers in Rural South India from a period of January 2021 to December 2021 was done. Medical records of 10,85,81 patients both new and reviewed, 79,562 newly registered patients and 29,019 review patient’s from15 Vision centers were included for data analysis. All the patients registered at the vision center underwent basic eye examination, including visual acuity, IOP measurement, Slit-lamp examination, retinoscopy, Fundus examination etc. Results: A total of 1,08,581 patients were included in the study. Of the total 1,08,581 patients, 79,562 were newly registered patients at Vision center and 29,019 were review patients. Males were 52,201(48.1%) and Females were 56,308(51.9) among them. The mean age of all examined patients was 41.03 ± 20.9 years (Standard deviation) and ranged from 01 – 113 years. Presenting mean visual acuity was 0.31 ± 0.5 in the right eye and 0.31 ± 0.4 in the left eye. Of the 1,08,581 patients 22,770 patients had refractive error in right eye and 22,721 patients had uncorrected refractive error in left eye. Glass prescription was given to 17,178 (15.8%) patients. 8,109 (7.5%) patients were referred to the base hospital for specialty clinic expert opinion or for cataract surgery. Conclusion: Vision center utilizing teleconsultation for comprehensive eye screening unit is a very effective tool in reducing the avoidable visual impairment caused due to uncorrected refractive error. Vision Centre model is believed to be efficient as it facilitates early detection and management of uncorrected refractive errors.

Keywords: refractive error, uncorrected refractive error, vision center, vision technician, teleconsultation

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Authors: Naveen Kumar Challa, Pavan Verıkıcherla, Madhubalan, Ashısh Sharma

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Aim: Aim of the study was to find whether any difference exists in pupil mydriasis measured with Orbscan in non-diabetic and type 2 diabetic patients at various intervals after installation of Tropicamide 0.8% and Phenylephrine 5%. Methods: the Observational study conducted at a tertiary care eye hospital during September 2014 to March 2015. 240 eyes from 120 patients (40 non-diabetic, 80 diabetic) were dilated with Tropicamide 0.8% and Phenylephrine 5%. One drop of a drug was installed twice. The second drop is installed at 20 minutes after installation of the first drop. In two groups’ pupil diameter was measured before installation of drops and also at 15, 30, 45 and 60 minutes after installation of the first drop using both Orbscan. Result: Mean age of the non-diabetic group is 48.67 ± 7.93 years; Diabetic group is 59.97 ± 8.77 years. Mean duration of Diabetes was 7.01 ± 5.05 years. Mean pupil diameter measured with Orbscan before installation of the drops and also at 15, 30, 45 and 60 minutes after installation of first drop in non-diabetic group was 4.18 ± 0.64mm, 6.15 ± 0.41mm, 7.76 ±0.34, 9.59 ± 0.30, and 9.97 ± 0.10 mm respectively and for the diabetic group it was 4.00 ± 0.56 mm, 5.53 ± 0.52 mm, 7.018 ± 0.58mm, 8.25±0.51mm and 9.18 ± 0.46mm respectively. The mean difference between the mean pupil diameters of the non-diabetic and diabetic group shows a significant difference (P< 0.01) at all intervals except before dilatation. There is a significant negative correlation (r = 0.78 – 0.92) between the duration of diabetes and pupil dilatation at all intervals after installation of the drops. There is also significant difference (P< 0.005) in the mean values of pupil diameter between non retinopathy diabetic subjects and diabetic retinopathy subjects at all intervals after installation of drops. Conclusion: People attending eye clinic, whose pupil mydriasis values falls below the normal may be referred for diabetic evaluation. If normative data is established for the pupil size in Indian population using Orbscan then the values fall under normative data could be a predictor for diabetes. This would in turn help ophthalmologist to detect the diabetes at an early stage and prevent the complications resulting from the diabetes.

Keywords: diabetes mellitus, pupil diameter, orbscan, tropicamide

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Authors: Kevin Kellner, Nga Lao, Orla Coleman, Paula Meleady, Niall Barron

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Chinese Hamster Ovary (CHO) cells are the prominent cell line used in biopharmaceutical production. To improve yields and find beneficial bioprocess phenotypes genetic engineering plays an essential role in recent research. The miR-23 cluster, specifically miR-24 and miR-27, was first identified as differentially expressed during hypothermic conditions suggesting a role in proliferation and productivity in CHO cells. In this study, we used sponge decoy technology to stably deplete the miRNA expression of the cluster. Furthermore, we implemented the CRISPR/Cas9 system to knockdown miRNA expression. Sponge constructs were designed for an imperfect binding of the miRNA target, protecting from RISC mediated cleavage. GuideRNAs for the CRISPR/Cas9 system were designed to target the seed region of the miRNA. The expression of mature miRNA and precursor were confirmed using RT-qPCR. For both approaches stable expressing mixed populations were generated and characterised in batch cultures. It was shown, that CRISPR/Cas9 can be implemented in CHO cells with achieving high knockdown efficacy of every single member of the cluster. Targeting of one miRNA member showed that its genomic paralog is successfully targeted as well. The stable depletion of miR-24 using CRISPR/Cas9 showed increased growth and specific productivity in a CHO-K1 mAb expressing cell line. This phenotype was further characterized using quantitative label-free LC-MS/MS showing 186 proteins differently expressed with 19 involved in proliferation and 26 involved in protein folding/translation. Targeting miR-27 in the same cell line showed increased viability in late stages of the culture compared to the control. To evaluate the phenotype in an industry relevant cell line; the miR-23 cluster, miR-24 and miR-27 were stably depleted in a Fc fusion CHO-S cell line which showed increased batch titers up to 1.5-fold. In this work, we highlighted that the stable depletion of the miR-23 cluster and its members can improve the bioprocess phenotype concerning growth and productivity in two different cell lines. Furthermore, we showed that using CRISPR/Cas9 is comparable to the traditional sponge decoy technology.

Keywords: Chinese Hamster ovary cells, CRISPR/Cas9, microRNAs, sponge decoy technology

Procedia PDF Downloads 180