Search results for: diagnostic feedback

Authors: Elaine Crable, Amit Sen

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Short-term study abroad programs have become a mainstay of MBA programs. The benefits of international business experiences, along with its exposure to global cultures, are well documented. However, developing a rewarding study, abroad program at the graduate level can be challenging for Faculty, especially when devising such a program for a group of part-time MBA students who come with a wide range of experiences and demographic characteristics. Each student has individual expectations for the study abroad experience. This study provides suggestions and considerations for Faculty that are planning to design a short-term study abroad program, especially for part-time MBA students. Insights are based on a recent experience leading a group of twenty-one students on a ten-day program to Morocco. The trip was designed and facilitated by two faculty members and a local Moroccan facilitator. This experience led to a number of insights and recommendations. First, the choice of location is critical. The choice of Morocco was very deliberate, owing to its multi-faceted cultural landscape and international business interest. It is an Islamic State with close ties to Europe both culturally and geographically and Morocco is a multi-lingual country with some combination of three languages spoken by most – English, Arabic, and French. Second, collaboration with a local ‘academic’ partner allowed the level of instruction to be both rigorous and significantly more engaging. Third, allowing students to participate in the planning of the trip enabled the trip participants to collaborate, negotiate, and share their own experiences and strengths. The pre-trip engagement was structured by creating four sub-groups, each responsible for an assigned city. Each student sub-group had to provide a historical background of the assigned city, plan the itinerary including sites to visit, cuisine to experience, industries to explore, markets to visit, plus provide a budget for that city’s expenses. The pre-planning segment of the course was critical for the success of the program as students were able to contribute to the design of the program through collaboration and negotiation with their peers. Fourth, each student sub-group was assigned industry to study within Morocco. The student sub-group prepared a presentation and a group paper with their analysis of the chosen industries. The pre-planning activities created strong bonds among the trip participants, which was evident when faced with on-ground challenges, especially when it was necessary to quickly evacuate due to a surprise USA COVID evacuation notice. The entire group supported each other when quickly making their way back to the United States. Unfortunately, the trip was cut short by two days due to this emergency exit, but the feedback regarding the program was very positive all around. While the program design put pressure on the Faculty leads regarding planning and coordination upfront, the outcome in terms of student engagement, student learning, collaboration and negotiation were all favorable and worth the effort. Finally, an added value, the cost of the program for the student was significantly lower compared to running a program with a professional provider.

Keywords: business education, experiential learning, international education, study abroad

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Authors: Pawel Martynowicz

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Vibrations are a crucial problem for slender structures such as towers, masts, chimneys, wind turbines, bridges, high buildings, etc., that is why most of them are equipped with vibration attenuation or fatigue reduction solutions. In this work, a slender structure (i.e., wind turbine tower-nacelle model) equipped with nonlinear, semiactive tuned vibration absorber(s) is analyzed. For this study purposes, magnetorheological (MR) dampers are used as semiactive actuators. Several optimal-based approaches to structural vibration attenuation are investigated against the standard ‘ground-hook’ law and passive tuned vibration absorber(s) implementations. The common approach to optimal control of nonlinear systems is offline computation of the optimal solution, however, so determined open loop control suffers from lack of robustness to uncertainties (e.g., unmodelled dynamics, perturbations of external forces or initial conditions), and thus perturbation control techniques are often used. However, proper linearization may be an issue for highly nonlinear systems with implicit relations between state, co-state, and control. The main contribution of the author is the development as well as numerical and experimental verification of the Pontriagin maximum-principle-based vibration control concepts that produce directly actuator control input (not the demanded force), thus force tracking algorithm that results in control inaccuracy is entirely omitted. These concepts, including one-step optimal control, quasi-optimal control, and optimal-based modified ‘ground-hook’ law, can be directly implemented in online and real-time feedback control for periodic (or semi-periodic) disturbances with invariant or time-varying parameters, as well as for non-periodic, transient or random disturbances, what is a limitation for some other known solutions. No offline calculation, excitations/disturbances assumption or vibration frequency determination is necessary, moreover, all of the nonlinear actuator (MR damper) force constraints, i.e., no active forces, lower and upper saturation limits, hysteresis-type dynamics, etc., are embedded in the control technique, thus the solution is optimal or suboptimal for the assumed actuator, respecting its limitations. Depending on the selected method variant, a moderate or decisive reduction in the computational load is possible compared to other methods of nonlinear optimal control, while assuring the quality and robustness of the vibration reduction system, as well as considering multi-pronged operational aspects, such as possible minimization of the amplitude of the deflection and acceleration of the vibrating structure, its potential and/or kinetic energy, required actuator force, control input (e.g. electric current in the MR damper coil) and/or stroke amplitude. The developed solutions are characterized by high vibration reduction efficiency – the obtained maximum values of the dynamic amplification factor are close to 2.0, while for the best of the passive systems, these values exceed 3.5.

Keywords: magnetorheological damper, nonlinear tuned vibration absorber, optimal control, real-time structural vibration attenuation, wind turbines

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Authors: Atefeh Omidkhah Kharashtomi, Rasoul Hedayat Nejad, Saeed Bakhtiyari

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Circulation paths and indoor connection network of the building play an important role both in the daily operation of the building and during evacuation in emergency situations. The degree of legibility of the paths for navigation inside the building has a deep connection with the perceptive and cognitive system of human, and the way the surrounding environment is being perceived. Human perception of the space is based on the sensory systems in a three-dimensional environment, and non-linearly, so it is necessary to avoid reducing its representations in architectural design as a two-dimensional and linear issue. Today, the advances in the field of virtual reality (VR) technology have led to various applications, and architecture and building science can benefit greatly from these capabilities. Especially in cases where the design solution requires a detailed and complete understanding of the human perception of the environment and the behavioral response, special attention to VR technologies could be a priority. Way-finding in the indoor circulation network is a proper example for such application. Success in way-finding could be achieved if human perception of the route and the behavioral reaction have been considered in advance and reflected in the architectural design. This paper discusses the VR technology applications for the way-finding improvements in indoor engineering of the building. In a systematic review, with a database consisting of numerous studies, firstly, four categories for VR applications for circulation way-finding have been identified: 1) data collection of key parameters, 2) comparison of the effect of each parameter in virtual environment versus real world (in order to improve the design), 3) comparing experiment results in the application of different VR devices/ methods with each other or with the results of building simulation, and 4) training and planning. Since the costs of technical equipment and knowledge required to use VR tools lead to the limitation of its use for all design projects, priority buildings for the use of VR during design are introduced based on case-studies analysis. The results indicate that VR technology provides opportunities for designers to solve complex buildings design challenges in an effective and efficient manner. Then environmental parameters and the architecture of the circulation routes (indicators such as route configuration, topology, signs, structural and non-structural components, etc.) and the characteristics of each (metrics such as dimensions, proportions, color, transparency, texture, etc.) are classified for the VR way-finding experiments. Then, according to human behavior and reaction in the movement-related issues, the necessity of scenario-based and experiment design for using VR technology to improve the design and receive feedback from the test participants has been described. The parameters related to the scenario design are presented in a flowchart in the form of test design, data determination and interpretation, recording results, analysis, errors, validation and reporting. Also, the experiment environment design is discussed for equipment selection according to the scenario, parameters under study as well as creating the sense of illusion in the terms of place illusion, plausibility and illusion of body ownership.

Keywords: virtual reality (VR), way-finding, indoor, circulation, design

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Authors: Denise De Fátima Fernandes Barbosa, Tyane Mayara Ferreira Oliveira, Diego Jorge Maia Lima, Paula Renata Amorim Lessa, Ana Karina Bezerra Pinheiro, Cintia Gondim Pereira Calou, Glauberto Da Silva Quirino, Hellen Lívia Oliveira Catunda, Tatiana Gomes Guedes, Nicolau Da Costa

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The aim of this study is to evaluate the use of Digital Cervicography (DC) in the diagnosis of precancerous lesions related to Human Papillomavirus (HPV). Cross-sectional study with a quantitative approach, of evaluative type, held in a health unit linked to the Pro Dean of Extension of the Federal University of Ceará, in the period of July to August 2015 with a sample of 33 women. Data collecting was conducted through interviews with enforcement tool. Franco (2005) standardized the technique used for DC. Polymerase Chain Reaction (PCR) was performed to identify high-risk HPV genotypes. DC were evaluated and classified by 3 judges. The results of DC and PCR were classified as positive, negative or inconclusive. The data of the collecting instruments were compiled and analyzed by the software Statistical Package for Social Sciences (SPSS) with descriptive statistics and cross-references. Sociodemographic, sexual and reproductive variables were analyzed through absolute frequencies (N) and their respective percentage (%). Kappa coefficient (κ) was applied to determine the existence of agreement between the DC of reports among evaluators with PCR and also among the judges about the DC results. The Pearson's chi-square test was used for analysis of sociodemographic, sexual and reproductive variables with the PCR reports. It was considered statistically significant (p<0.05). Ethical aspects of research involving human beings were respected, according to 466/2012 Resolution. Regarding the socio-demographic profile, the most prevalent ages and equally were those belonging to the groups 21-30 and 41-50 years old (24.2%). The brown color was reported in excess (84.8%) and 96.9% out of them had completed primary and secondary school or studying. 51.5% were married, 72.7% Catholic, 54.5% employed and 48.5% with income between one and two minimum wages. As for the sexual and reproductive characteristics, prevailed heterosexual (93.9%) who did not use condoms during sexual intercourse (72.7%). 51.5% had a previous history of Sexually Transmitted Infection (STI), and HPV the most prevalent STI (76.5%). 57.6% did not use contraception, 78.8% underwent examination Cancer Prevention Uterus (PCCU) with shorter time interval or equal to one year, 72.7% had no cases of Cervical Cancer in the family, 63.6% were multiparous and 97% were not vaccinated against HPV. DC identified good level of agreement between raters (κ=0.542), had a specificity of 77.8% and sensitivity of 25% when compared their results with PCR. Only the variable race showed a statistically significant association with CRP (p=0.042). DC had 100% acceptance amongst women in the sample, revealing the possibility of other experiments in using this method so that it proves as a viable technique. The DC positivity criteria were developed by nurses and these professionals also perform PCCU in Brazil, which means that DC can be an important complementary diagnostic method for the appreciation of these professional’s quality of examinations.

Keywords: gynecological examination, human papillomavirus, nursing, papillomavirus infections, uterine lasmsneop

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Authors: Marcus Goh Tian Xi, Alicia Chua Si Wen, Eunice Gan Ghee Wu, Helen Bound, Lee Liang Ying, Albert Lee

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The Map of Dialogical Inquiry provides a conceptual look at the underlying nature of future-oriented skills. According to the Map, learning is learner-oriented, with conversational time shifted from teachers to learners, who play a strong role in deciding what and how they learn. For example, in courses operating on the principles of Dialogical Inquiry, learners were able to leave the classroom with a deeper understanding of the topic, broader exposure to differing perspectives, and stronger critical thinking capabilities, compared to traditional approaches to teaching. Despite its contributions to learning, the Map is grounded in a qualitative approach both in its development and its application for providing feedback to learners and educators. Studies hinge on openended responses by Map users, which can be time consuming and resource intensive. The present research is motivated by this gap in practicality by aiming to develop and validate a quantitative measure of the Map. In addition, a quantifiable measure may also strengthen applicability by making learning experiences trackable and comparable. The Map outlines eight learning aspects that learners should holistically engage. This research focuses on the Analysing aspect of learning. According to the Map, Analysing has four key components: liking or engaging in logic, using interpretative lenses, seeking patterns, and critiquing and deconstructing. Existing scales of constructs (e.g., critical thinking, rationality) related to these components were identified so that the current scale could adapt items from. Specifically, items were phrased beginning with an “I”, followed by an action phrase, to fulfil the purpose of assessing learners' engagement with Analysing either in general or in classroom contexts. Paralleling standard scale development procedure, the 26-item Analysing scale was administered to 330 participants alongside existing scales with varying levels of association to Analysing, to establish construct validity. Subsequently, the scale was refined and its dimensionality, reliability, and validity were determined. Confirmatory factor analysis (CFA) revealed if scale items loaded onto the four factors corresponding to the components of Analysing. To refine the scale, items were systematically removed via an iterative procedure, according to their factor loadings and results of likelihood ratio tests at each step. Eight items were removed this way. The Analysing scale is better conceptualised as unidimensional, rather than comprising the four components identified by the Map, for three reasons: 1) the covariance matrix of the model specified for the CFA was not positive definite, 2) correlations among the four factors were high, and 3) exploratory factor analyses did not yield an easily interpretable factor structure of Analysing. Regarding validity, since the Analysing scale had higher correlations with conceptually similar scales than conceptually distinct scales, with minor exceptions, construct validity was largely established. Overall, satisfactory reliability and validity of the scale suggest that the current procedure can result in a valid and easy-touse measure for each aspect of the Map.

Keywords: analytical thinking, dialogical inquiry, education, lifelong learning, pedagogy, scale development

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Authors: Sibawu Witness Siyepu

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This study explores the effects of instructional design using blended learning in the learning of radian measures among Engineering students. Blended learning is an education programme that combines online digital media with traditional classroom methods. It requires the physical presence of both lecturer and student in a mathematics computer laboratory. Blended learning provides element of class control over time, place, path or pace. The focus was on the use of Khan Academy to supplement traditional classroom interactions. Khan Academy is a non-profit educational organisation created by educator Salman Khan with a goal of creating an accessible place for students to learn through watching videos in a computer assisted computer. The researcher who is an also lecturer in mathematics support programme collected data through instructing students to watch Khan Academy videos on radian measures, and by supplying students with traditional classroom activities. Classroom activities entails radian measure activities extracted from the Internet. Students were given an opportunity to engage in class discussions, social interactions and collaborations. These activities necessitated students to write formative assessments tests. The purpose of formative assessments tests was to find out about the students’ understanding of radian measures, including errors and misconceptions they displayed in their calculations. Identification of errors and misconceptions serve as pointers of students’ weaknesses and strengths in their learning of radian measures. At the end of data collection, semi-structure interviews were administered to a purposefully sampled group to explore their perceptions and feedback regarding the use of blended learning approach in teaching and learning of radian measures. The study employed Algebraic Insight Framework to analyse data collected. Algebraic Insight Framework is a subset of symbol sense which allows a student to correctly enter expressions into a computer assisted systems efficiently. This study offers students opportunities to enter topics and subtopics on radian measures into a computer through the lens of Khan Academy. Khan academy demonstrates procedures followed to reach solutions of mathematical problems. The researcher performed the task of explaining mathematical concepts and facilitated the process of reinvention of rules and formulae in the learning of radian measures. Lastly, activities that reinforce students’ understanding of radian were distributed. Results showed that this study enthused the students in their learning of radian measures. Learning through videos prompted the students to ask questions which brought about clarity and sense making to the classroom discussions. Data revealed that sense making through reinvention of rules and formulae assisted the students in enhancing their learning of radian measures. This study recommends the use of Khan Academy in blended learning to be introduced as a socialisation programme to all first year students. This will prepare students that are computer illiterate to become conversant with the use of Khan Academy as a powerful tool in the learning of mathematics. Khan Academy is a key technological tool that is pivotal for the development of students’ autonomy in the learning of mathematics and that promotes collaboration with lecturers and peers.

Keywords: algebraic insight framework, blended learning, Khan Academy, radian measures

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Authors: Marielle Leijten, Catherine Meulemans, Sven De Maeyer, Luuk Van Waes

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For the diagnosis of Alzheimer's disease (AD), a large variety of neuropsychological tests are available. In some of these tests, linguistic processing - both oral and written - is an important factor. Language disturbances might serve as a strong indicator for an underlying neurodegenerative disorder like AD. However, the current diagnostic instruments for language assessment mainly focus on product measures, such as text length or number of errors, ignoring the importance of the process that leads to written or spoken language production. In this study, it is our aim to describe and test differences between cognitive and impaired elderly on the basis of a selection of writing process variables (inter- and intrapersonal characteristics). These process variables are mainly related to pause times, because the number, length, and location of pauses have proven to be an important indicator of the cognitive complexity of a process. Method: Participants that were enrolled in our research were chosen on the basis of a number of basic criteria necessary to collect reliable writing process data. Furthermore, we opted to match the thirteen cognitively impaired patients (8 MCI and 5 AD) with thirteen cognitively healthy elderly. At the start of the experiment, participants were each given a number of tests, such as the Mini-Mental State Examination test (MMSE), the Geriatric Depression Scale (GDS), the forward and backward digit span and the Edinburgh Handedness Inventory (EHI). Also, a questionnaire was used to collect socio-demographic information (age, gender, eduction) of the subjects as well as more details on their level of computer literacy. The tests and questionnaire were followed by two typing tasks and two picture description tasks. For the typing tasks participants had to copy (type) characters, words and sentences from a screen, whereas the picture description tasks each consisted of an image they had to describe in a few sentences. Both the typing and the picture description tasks were logged with Inputlog, a keystroke logging tool that allows us to log and time stamp keystroke activity to reconstruct and describe text production processes. The main rationale behind keystroke logging is that writing fluency and flow reveal traces of the underlying cognitive processes. This explains the analytical focus on pause (length, number, distribution, location, etc.) and revision (number, type, operation, embeddedness, location, etc.) characteristics. As in speech, pause times are seen as indexical of cognitive effort. Results. Preliminary analysis already showed some promising results concerning pause times before, within and after words. For all variables, mixed effects models were used that included participants as a random effect and MMSE scores, GDS scores and word categories (such as determiners and nouns) as a fixed effect. For pause times before and after words cognitively impaired patients paused longer than healthy elderly. These variables did not show an interaction effect between the group participants (cognitively impaired or healthy elderly) belonged to and word categories. However, pause times within words did show an interaction effect, which indicates pause times within certain word categories differ significantly between patients and healthy elderly.

Keywords: Alzheimer's disease, keystroke logging, matching, writing process

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Authors: Kuo-Kai Lin, Po-Lun Chang

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Research Background and Purpose: Following the development of the Internet and multimedia, the Internet and information technology have become crucial avenues of modern communication and knowledge acquisition. The advantages of using mobile devices for learning include making learning borderless and accessible. Mobile learning has become a trend in disease management and health promotion in recent years. End-stage renal disease (ESRD) is an irreversible chronic disease, and patients who do not receive kidney transplants can only rely on hemodialysis or peritoneal dialysis to survive. Due to the complexities in caregiving for patients with ESRD that stem from their advanced age and other comorbidities, the patients’ incapacity of self-care leads to an increase in the need to rely on their families or primary caregivers, although whether the primary caregivers adequately understand and implement patient care is a topic of concern. Therefore, this study explored whether primary caregivers’ health care provisions can be improved through the intervention of an automatic speech intelligent system, thereby improving the objective health outcomes of patients undergoing long-term dialysis. Method: This study developed an automatic speech intelligent system with healthcare functions such as health information voice prompt, two-way feedback, real-time push notification, and health information delivery. Convenience sampling was adopted to recruit eligible patients from a hemodialysis center at a regional teaching hospital as research participants. A one-group pretest-posttest design was adopted. Descriptive and inferential statistics were calculated from the demographic information collected from questionnaires answered by patients and primary caregivers, and from a medical record review, a health care scale (recorded six months before and after the implementation of intervention measures), a subjective health assessment, and a report of objective physiological indicators. The changes in health care behaviors, subjective health status, and physiological indicators before and after the intervention of the proposed automatic speech intelligent system were then compared. Conclusion and Discussion: The preliminary automatic speech intelligent system developed in this study was tested with 20 pretest patients at the recruitment location, and their health care capacity scores improved from 59.1 to 72.8; comparisons through a nonparametric test indicated a significant difference (p < .01). The average score for their subjective health assessment rose from 2.8 to 3.3. A survey of their objective physiological indicators discovered that the compliance rate for the blood potassium level was the most significant indicator; its average compliance rate increased from 81% to 94%. The results demonstrated that this automatic speech intelligent system yielded a higher efficacy for chronic disease care than did conventional health education delivered by nurses. Therefore, future efforts will continue to increase the number of recruited patients and to refine the intelligent system. Future improvements to the intelligent system can be expected to enhance its effectiveness even further.

Keywords: automatic speech intelligent system for health care, primary caregiver, long-term hemodialysis, health care capabilities, health outcomes

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Authors: Alaa Abdellatif, Gabrièle Breda

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Background. Advanced therapy medicinal products (ATMPs) are innovative therapies that mainly target orphan diseases and high unmet medical needs. ATMP includes gene therapy medicinal products (GTMP), somatic cell therapy medicinal products (CTMP), and tissue-engineered therapies (TEP). Since legislation opened the way in 2007, 25 ATMPs have been approved in the EU, which is about the same amount as the U.S. Food and Drug Administration. However, not all of the ATMPs that have been approved have successfully reached the market and retained their approval. Objectives. We aim to understand all the factors limiting the market access to very promising therapies in a systemic approach, to be able to overcome these problems, in the future, with scientific, regulatory and commercial innovations. Further to recent reviews that focus either on specific countries, products, or dimensions, we will address all the challenges faced by ATMP development today. Methodology. We used mixed methods and a multi-level approach for data collection. First, we performed an updated academic literature review on ATMP development and their scientific and market access challenges (papers published between 2018 and April 2023). Second, we analyzed industry feedback from cell and gene therapy webinars and white papers published by providers and pharmaceutical industries. Finally, we established a comparative analysis of the regulatory guidelines published by EMA and the FDA for ATMP approval. Results: The main challenges in bringing these therapies to market are the high development costs. Developing ATMPs is expensive due to the need for specialized manufacturing processes. Furthermore, the regulatory pathways for ATMPs are often complex and can vary between countries, making it challenging to obtain approval and ensure compliance with different regulations. As a result of the high costs associated with ATMPs, challenges in obtaining reimbursement from healthcare payers lead to limited patient access to these treatments. ATMPs are often developed for orphan diseases, which means that the patient population is limited for clinical trials which can make it challenging to demonstrate their safety and efficacy. In addition, the complex manufacturing processes required for ATMPs can make it challenging to scale up production to meet demand, which can limit their availability and increase costs. Finally, ATMPs face safety and efficacy challenges: dangerous adverse events of these therapies like toxicity related to the use of viral vectors or cell therapy, starting material and donor-related aspects. Conclusion. As a result of our mixed method analysis, we found that ATMPs face a number of challenges in their development, regulatory approval, and commercialization and that addressing these challenges requires collaboration between industry, regulators, healthcare providers, and patient groups. This first analysis will help us to address, for each challenge, proper and innovative solution(s) in order to increase the number of ATMPs approved and reach the patients

Keywords: advanced therapy medicinal products (ATMPs), product development, market access, innovation

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Authors: Jenee Gooden, Kevin Vasquez-monterroso, Lady Paula Dejesus, Sandra Wainwright, Daniel Kim, Mackenzie Walker

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Introduction: Pyoderma gangrenosum (PG) is a rare, rapidly progressive, neutrophilic ulcerative colitis condition with an incidence of 3 to 10 cases per year ¹ ². Due to the similar appearance, PG is often misdiagnosed as a diabetic ulcer in diabetic patients. Though they may clinically appear similar in appearance, the treatment protocol and diagnostic criteria differ. Also, end-stage renal disease (ESRD) is often a condition seen in diabetic patients, which can have a significant impact on wound healing due to the wide range of uremic toxins³. This case study demonstrates a multidisciplinary team and multimodal treatment approach by podiatric surgery, general surgery, rheumatology, infectious disease, interventional cardiology, wound care and hyperbaric medicine for an uncontrolled diabetic with pyoderma gangrenosum of a significant circumferential wound, covering almost the entire right lower extremity. Methods:56 y.o male presents with multiple PG ulcerations, including the chest, right posterior lower extremity and sacrum. All ulcerations were previously managed by the same wound care specialist. His chief complaint was worsening PG ulcerations accompanied by a fever of 103 °F . This case study focuses on the wound to his RLE. Past medical history significant for diabetes mellitus type 2 with hemoglobin A1c of 10% and end stage renal disease (ESRD) on hemodialysis. A multidisciplinary team approach by podiatric surgery, general surgery, rheumatology, infectious disease, interventional cardiology, wound care and hyperbaric medicine was successfully used to perform right lower extremity limb salvage. The patient was managed by rheumatology for the continuation of prior medication, as well as the mutual agreement with wound care for the addition of dapsone. A coronary CT angiogram was performed by interventional cardiology, but no significant disease was noted, and no further vascular workup was necessary. Multiple surgical sharp wide excisional debridements with application of allografts and split thickness skin grafts for the circumferential ulceration that encompassed almost the entire right lower extremity were performed by both podiatric surgery and general surgery. Wound cultures and soft tissue biopsies were performed, and infectious disease managed antibiotic therapy. Hyperbaric oxygen therapy and wound vac therapy by wound care were also completed as adjunct management. Results: Prevention of leg amputation by limb salvage of the RLE was accomplished by a multidisciplinary team approach, with the wound size decreasing over a total of 29 weeks from 600 cm² to 12.0 x 3.5 x 0.2 cm. Our multidisciplinary team included podiatric surgery, general surgery, rheumatology, infectious disease, interventional cardiology, wound care and hyperbaric medicine. Discussion: Wound healing, in general, can have its challenges, and those challenges are only magnified when accompanied by multiple systemic illnesses. Though the negative impact of diabetes on wound healing is well known, the compound impact of being a diabetic with ESRD and having pyoderma gangrenosum is not. This case demonstrates the necessity for a multidisciplinary team approach with a wide array of treatment modalities to optimize wound healing and perform limb salvage with prevention of lower extremity amputation.

Keywords: diabetes, podiatry, pyoderma gangrenosum, end stage renal disease

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Authors: Lauren G. Coggins

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Catheter-associated urinary tract infections (CAUTI) and central line-associated bloodstream infections (CLABSI) are among the most common healthcare-associated infections (HAI), contributing to prolonged lengths of stay, greater costs of patient care, and increased patient mortality. Evidence-based preventative care bundles exist to establish consistent, safe patient-care practices throughout an entire organization, helping to ensure the collective application of care strategies that aim to improve patient outcomes and minimize complications. The cardiac intensive care unit at a nationally ranked teaching and research hospital in the United States exceeded its annual CAUTI and CLABSI targets in the fiscal year 2019, prompting examination into the unit’s infection prevention efforts that included preventative care bundles for both HAIs. Adherence to the CAUTI and CLABSI preventative care bundles was evaluated through frequent audits conducted over three months, using standards and resources from The Joint Commission, a globally recognized leader in quality improvement in healthcare and patient care safety. The bundle elements with the lowest scores were identified as the most commonly missed elements. Three elements from both bundles, six elements in total, served as key content areas for the educational interventions targeted to bedside nurses. The CAUTI elements included appropriate urinary catheter order, appropriate continuation criteria, and urinary catheter care. The CLABSI elements included primary tubing compliance, needleless connector compliance, and dressing change compliance. An integrated, multi-platform education campaign featured content on each CAUTI and CLABSI preventative care bundle in its entirety, with additional reinforcement focused on the lowest scoring elements. One-on-one educational materials included an informational pamphlet, badge buddy, a presentation to reinforce nursing care standards, and real-time application through case studies and electronic health record demonstrations. A digital hub was developed on the hospital’s Intranet for quick access to unit resources, and a bulletin board helped track the number of days since the last CAUTI and CLABSI incident. Audits continued to be conducted throughout the education campaign, and staff were given real-time feedback to address any gaps in adherence. Nearly every nurse in the cardiac intensive care unit received all educational materials, and adherence to all six key bundle elements increased after the implementation of educational interventions. Recommendations from this implementation include providing consistent, comprehensive education across multiple teaching tools and regular audits to track adherence. The multi-platform education campaign brought focus to the evidence-based CAUTI and CLABSI bundles, which in turn will help to reduce CAUTI and CLABSI rates in clinical practice.

Keywords: education, healthcare-associated infections, infection, nursing, prevention

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Authors: Adiba Matin, Daniel Cabrera, Javiera Bellolio, Jasmine Stewart, Dana Gerberi (librarian), Nathan Cummins, Fernanda Bellolio

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OBJECTIVES: Artificial intelligence (AI) tools are becoming more prevalent in healthcare settings, particularly for diagnostic and therapeutic recommendations, with an expected surge in the incoming years. The bedside use of this technology for clinicians opens the possibility of disagreements between the recommendations from AI algorithms and clinicians’ judgment. There is a paucity in the literature analyzing nature and possible outcomes of these potential conflicts, particularly related to ethical considerations. The goal of this scoping review is to identify, analyze and classify current themes and potential strategies addressing ethical conflicts originating from the conflict between AI and human recommendations. METHODS: A protocol was written prior to the initiation of the study. Relevant literature was searched by a medical librarian for the terms of artificial intelligence, healthcare and liability, ethics, or conflict. Search was run in 2021 in Ovid Cochrane Central Register of Controlled Trials, Embase, Medline, IEEE Xplore, Scopus, and Web of Science Core Collection. Articles describing the role of AI in healthcare that mentioned conflict between humans and AI were included in the primary search. Two investigators working independently and in duplicate screened titles and abstracts and reviewed full-text of potentially eligible studies. Data was abstracted into tables and reported by themes. We followed methodological guidelines for Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR). RESULTS: Of 6846 titles and abstracts, 225 full texts were selected, and 48 articles included in this review. 23 articles were included as original research and review papers. 25 were included as editorials and commentaries with similar themes. There was a lack of consensus in the included articles on who would be held liable for mistakes incurred by following AI recommendations. It appears that there is a dichotomy of the perceived ethical consequences depending on if the negative outcome is a result of a human versus AI conflict or secondary to a deviation from standard of care. Themes identified included transparency versus opacity of recommendations, data bias, liability of outcomes, regulatory framework, and the overall scope of artificial intelligence in healthcare. A relevant issue identified was the concern by clinicians of the “black box” nature of these recommendations and the ability to judge appropriateness of AI guidance. CONCLUSION AI clinical tools are being rapidly developed and adopted, and the use of this technology will create conflicts between AI algorithms and healthcare workers with various outcomes. In turn, these conflicts may have legal, and ethical considerations. There is limited consensus about the focus of ethical and liability for outcomes originated from disagreements. This scoping review identified the importance of framing the problem in terms of conflict between standard of care or not, and informed by the themes of transparency/opacity, data bias, legal liability, absent regulatory frameworks and understanding of the technology. Finally, limited recommendations to mitigate ethical conflicts between AI and humans have been identified. Further work is necessary in this field.

Keywords: ethics, artificial intelligence, emergency medicine, review

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Authors: Lakshmi Rao Kandukuri, Mamata Deenadayal, Suma Prasad, Bipin Sethi, Srinadh Buragadda, Lalji Singh

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Worldwide; at least 7.6 million children are born annually with severe genetic or congenital malformations and among them 90% of these are born in mid and low-income countries. Precise prevalence data are difficult to collect, especially in developing countries, owing to the great diversity of conditions and also because many cases remain undiagnosed. The genetic and congenital disorder is the second most common cause of infant and childhood mortality and occurs with a prevalence of 25-60 per 1000 births. The higher prevalence of genetic diseases in a particular community may, however, be due to some social or cultural factors. Such factors include the tradition of consanguineous marriage, which results in a higher rate of autosomal recessive conditions including congenital malformations, stillbirths, or mental retardation. Genetic diseases can vary in severity, from being fatal before birth to requiring continuous management; their onset covers all life stages from infancy to old age. Those presenting at birth are particularly burdensome and may cause early death or life-long chronic morbidity. Genetic testing for several genetic diseases identifies changes in chromosomes, genes, or proteins. The results of a genetic test can confirm or rule out a suspected genetic condition or help determine a person's chance of developing or passing on a genetic disorder. Several hundred genetic tests are currently in use and more are being developed. Chromosomal abnormalities are the major cause of human suffering, which are implicated in mental retardation, congenital malformations, dysmorphic features, primary and secondary amenorrhea, reproductive wastage, infertility neoplastic diseases. Cytogenetic evaluation of patients is helpful in the counselling and management of affected individuals and families. We present here especially chromosomal abnormalities which form a major part of genetic disease burden in India. Different programmes on chromosome research and human reproductive genetics primarily relate to infertility since this is a major public health problem in our country, affecting 10-15 percent of couples. Prenatal diagnosis of chromosomal abnormalities in high-risk pregnancies helps in detecting chromosomally abnormal foetuses. Such couples are counselled regarding the continuation of pregnancy. In addition to the basic research, the team is providing chromosome diagnostic services that include conventional and advanced techniques for identifying various genetic defects. Other than routine chromosome diagnosis for infertility, also include patients with short stature, hypogonadism, undescended testis, microcephaly, delayed developmental milestones, familial, and isolated mental retardation, and cerebral palsy. Thus, chromosome diagnostics has found its applicability not only in disease prevention and management but also in guiding the clinicians in certain aspects of treatment. It would be appropriate to affirm that chromosomes are the images of life and they unequivocally mirror the states of human health. The importance of genetic counseling is increasing with the advancement in the field of genetics. The genetic counseling can help families to cope with emotional, psychological, and medical consequences of genetic diseases.

Keywords: India, chromosome abnormalities, genetic disorders, cytogenetic study

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Authors: Claudia Mertens, Amanda Shufflebarger

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Step one: What we know about effective learning for pupils with SEN: results of 2 systematic reviews: Before establishing principles and practices for teaching and learning of pupils with SEN, we need a good overview of the results of empirical studies conducted in the respective field. Therefore, two systematic reviews on the use of digital tools in inclusive and non-inclusive school settings were conducted - taking into consideration studies published in German: One systematic review included studies having undergone a peer review process, and the second included studies without peer review). The results (collaboration of two German universities) will be presented during the conference. Step two: Students’ results of a research lab on “inclusive media education”: On this basis, German students worked on “inclusive media education” in small research projects (duration: 1 year). They were “education majors” enrolled in a course on inclusive media education. They conducted research projects on topics ranging from smartboards in inclusive settings, digital media in gifted math education, Tik Tok in German as a Foreign Language education and many more. As part of their course, the German students created an academic conference poster. In the conference, the results of these research projects/papers are put into the context of the results of the systematic reviews. Step three: Global Classroom: The German students’ posters were critically discussed in a global classroom in cooperation with Indiana University East (USA) and Hamburg University (Germany) in the winter/spring term of 2022/2023. 15 students in Germany collaborated with 15 students at Indiana University East. The IU East student participants were enrolled in “Writing in the Arts and Sciences,” which is specifically designed for pre-service teachers. The joint work began at the beginning of the Spring 2023 semester in January 2023 and continued until the end of the Uni Hamburg semester in February 2023. Before January, Uni Hamburg students had been working on a research project individually or in pairs. Didactic Approach: Both groups of students posted a brief video or audio introduction to a shared Canvas discussion page. In the joint long synchronous session, the students discussed key content terms such as inclusion, inclusive, diversity, etc., with the help of prompt cards, and they compared how they understood or applied these terms differently. Uni Hamburg students presented drafts of academic posters. IU East students gave them specific feedback. After that, IU East students wrote brief reflections summarizing what they learned from the poster. After the class, small groups were expected to create a voice recording reflecting on their experiences. In their recordings, they examined critical incidents, highlighting what they learned from these incidents. Major results of the student research and of the global classroom collaboration can be highlighted during the conference. Results: The aggregated results of the two systematic reviews AND of the research lab/global classroom can now be a sound basis for 1) improving accessibility for students with SEN and 2) for adjusting teaching materials and concepts to the needs of the students with SEN - in order to create successful learning.

Keywords: digitalization, inclusion, inclusive media education, global classroom, systematic review

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Authors: Teresita Lopez Joy, Jose A. Acevedo Suarez, Martha I. Gomez Acosta, Ana Julia Acevedo Urquiaga

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Current trends in business competitiveness indicate the need to manage businesses as supply chains and not in isolation. The use of strategies aimed at maximum satisfaction of customers in a network and based on inter-company cooperation; contribute to obtaining successful joint results. In the Cuban economic context, the development of productive linkages to achieve integrated management of supply chains is considering a key aspect. In order to achieve this jump, it is necessary to develop acting capabilities in the entities that make up the chains through a systematic procedure that allows arriving at a management model in consonance with the environment. The objective of the research focuses on: designing a model and procedure for the development of integrated management of supply chains in economic entities. The results obtained are: the Model and the Procedure for the Development of the Supply Chains Integrated Management (MP-SCIM). The Model is based on the development of logistics in the network actors, the joint work between companies, collaborative planning and the monitoring of a main indicator according to the end customers. The application Procedure starts from the well-founded need for development in a supply chain and focuses on training entrepreneurs as doers. The characterization and diagnosis is done to later define the design of the network and the relationships between the companies. It takes into account the feedback as a method of updating the conditions and way to focus the objectives according to the final customers. The MP-SCIM is the result of systematic work with a supply chain approach in companies that have consolidated as coordinators of their network. The cases of the edible oil chain and explosives for construction sector reflect results of more remarkable advances since they have applied this approach for more than 5 years and maintain it as a general strategy of successful development. The edible oil trading company experienced a jump in sales. In 2006, the company started the analysis in order to define the supply chain, apply diagnosis techniques, define problems and implement solutions. The involvement of the management and the progressive formation of performance capacities in the personnel allowed the application of tools according to the context. The company that coordinates the explosives chain for construction sector shows adequate training with independence and opportunity in the face of different situations and variations of their business environment. The appropriation of tools and techniques for the analysis and implementation of proposals is a characteristic feature of this case. The coordinating entity applies integrated supply chain management to its decisions based on the timely training of the necessary action capabilities for each situation. Other cases of study and application that validate these tools are also detailed in this paper, and they highlight the results of generalization in the quantitative and qualitative improvement according to the final clients. These cases are: teaching literature in universities, agricultural products of local scope and medicine supply chains.

Keywords: integrated management, logistic system, supply chain management, tactical-operative planning

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Authors: Rania Raouf Sedky

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Adaptive architecture relates to buildings specifically designed to adapt to their residents and their environments. To design a biologically adaptive system, we can observe how living creatures in nature constantly adapt to different external and internal stimuli to be a great inspiration. The issue is not just how to create a system that is capable of change but also how to find the quality of change and determine the incentive to adapt. The research examines the possibilities of transforming spaces using the human body as an active tool. The research also aims to design and build an effective dynamic structural system that can be applied on an architectural scale and integrate them all into the creation of a new adaptive system that allows us to conceive a new way to design, build and experience architecture in a dynamic manner. The main objective was to address the possibility of a reciprocal transformation between the user and the architectural element so that the architecture can adapt to the user, as the user adapts to architecture. The motivation is the desire to deal with the psychological benefits of an environment that can respond and thus empathize with human emotions through its ability to adapt to the user. Adaptive affiliations of kinematic structures have been discussed in architectural research for more than a decade, and these issues have proven their effectiveness in developing kinematic structures, responsive and adaptive, and their contribution to 'smart architecture'. A wide range of strategies have been used in building complex kinetic and robotic systems mechanisms to achieve convertibility and adaptability in engineering and architecture. One of the main contributions of this research is to explore how the physical environment can change its shape to accommodate different spatial displays based on the movement of the user’s body. The main focus is on the relationship between materials, shape, and interactive control systems. The intention is to develop a scenario where the user can move, and the structure interacts without any physical contact. The soft form of shifting language and interaction control technology will provide new possibilities for enriching human-environmental interactions. How can we imagine a space in which to construct and understand its users through physical gestures, visual expressions, and response accordingly? How can we imagine a space whose interaction depends not only on preprogrammed operations but on real-time feedback from its users? The research also raises some important questions for the future. What would be the appropriate structure to show physical interaction with the dynamic world? This study concludes with a strong belief in the future of responsive motor structures. We imagine that they are developing the current structure and that they will radically change the way spaces are tested. These structures have obvious advantages in terms of energy performance and the ability to adapt to the needs of users. The research highlights the interface between remote sensing and a responsive environment to explore the possibility of an interactive architecture that adapts to and responds to user movements. This study ends with a strong belief in the future of responsive motor structures. We envision that it will improve the current structure and that it will bring a fundamental change to the way in which spaces are tested.

Keywords: adaptive architecture, interactive architecture, responsive architecture, tensegrity

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Authors: B. Godefroy, D. Beladjine, K. Beddiar

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In France, construction BIM actors commonly evoke the BIM gains for exploitation by integrating of the life cycle of a building. The standardization of level 7 of development would achieve this stage of the digital model. The householders include local public authorities, social landlords, public institutions (health and education), enterprises, facilities management companies. They have a dual role: owner and manager of their housing complex. In a context of financial constraint, the BIM of exploitation aims to control costs, make long-term investment choices, renew the portfolio and enable environmental standards to be met. It assumes a knowledge of the existing buildings, marked by its size and complexity. The information sought must be synthetic and structured, it concerns, in general, a real estate complex. We conducted a study with professionals about their concerns and ways to use it to see how householders could benefit from this development. To obtain results, we had in mind the recurring interrogation of the project management, on the needs of the operators, we tested the following stages: 1) Inculcate a minimal culture of BIM with multidisciplinary teams of the operator then by business, 2) Learn by BIM tools, the adaptation of their trade in operations, 3) Understand the place and creation of a graphic and technical database management system, determine the components of its library so their needs, 4) Identify the cross-functional interventions of its managers by business (operations, technical, information system, purchasing and legal aspects), 5) Set an internal protocol and define the BIM impact in their digital strategy. In addition, continuity of management by the integration of construction models in the operation phase raises the question of interoperability in the control of the production of IFC files in the operator’s proprietary format and the export and import processes, a solution rivaled by the traditional method of vectorization of paper plans. Companies that digitize housing complex and those in FM produce a file IFC, directly, according to their needs without recourse to the model of construction, they produce models business for the exploitation. They standardize components, equipment that are useful for coding. We observed the consequences resulting from the use of the BIM in the property industry and, made the following observations: a) The value of data prevail over the graphics, 3D is little used b) The owner must, through his organization, promote the feedback of technical management information during the design phase c) The operator's reflection on outsourcing concerns the acquisition of its information system and these services, observing the risks and costs related to their internal or external developments. This study allows us to highlight: i) The need for an internal organization of operators prior to a response to the construction management ii) The evolution towards automated methods for creating models dedicated to the exploitation, a specialization would be required iii) A review of the communication of the project management, management continuity not articulating around his building model, it must take into account the environment of the operator and reflect on its scope of action.

Keywords: information system, interoperability, models for exploitation, property industry

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Authors: Nelomi Anandagoda, Leanne J. Eveson

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During the COVID-19 pandemic, the surge in in-patient admissions across the medical directorate of a district general hospital necessitated the implementation of an incident rota. Conscious of the impact on training and professional development, the idea of developing a virtual teaching programme was conceived. The programme initially aimed to provide junior doctors, specialist nurses, pharmacists, and allied healthcare professionals from medical specialties and those re-deployed from other specialties (e.g., ophthalmology, GP, surgery, psychiatry) the knowledge and skills to manage the deteriorating patient with COVID-19. The programme was later developed to incorporate the general internal medicine curriculum. To facilitate continuing medical education whilst maintaining social distancing during this period, a virtual platform was used to deliver teaching to junior doctors across two large district general hospitals and two community hospitals. Teaching sessions were recorded and uploaded to a common platform, providing a resource for participants to catch up on and re-watch teaching sessions, making strides towards reducing discrimination against the professional development of less than full-time trainees. Thus, creating a learning environment, which is inclusive and accessible to adult learners in a self-directed manner. The negative impact of the pandemic on the well-being of healthcare professionals is well documented. To support the multi-disciplinary team, the virtual teaching programme evolved to included sessions on well-being, resilience, and work-life balance. Providing teaching for learners across the multi-disciplinary team (MDT) has been an eye-opening experience. By challenging the concept that learners should only be taught within their own peer groups, the authors have fostered a greater appreciation of the strengths of the MDT and showcased the immense wealth of expertise available within the trust. The inclusive nature of the teaching and the ease of joining a virtual teaching session has facilitated the dissemination of knowledge across the MDT, thus improving patient care on the frontline. The weekly teaching programme has been running for over eight months, with ongoing engagement, interest, and participation. As described above, the teaching programme has evolved to accommodate the needs of its learners. It has received excellent feedback with an appreciation of its inclusive, multi-disciplinary, and holistic nature. The COVID-19 pandemic provided a catalyst to rapidly develop novel methods of working and training and widened access/exposure to the virtual technologies available to large organisations. By merging pedagogical expertise and technology, the authors have created an effective online learning environment. Although the authors do not propose to replace face-to-face teaching altogether, this model of virtual multidisciplinary team, cross-site teaching has proven to be a great leveler. It has made high-quality teaching accessible to learners of different confidence levels, grades, specialties, and working patterns.

Keywords: cross-site, cross-speciality, inter-disciplinary, multidisciplinary, virtual teaching

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Authors: Prajakta Patil, Yash Huzurbazar, Abhijeet Shinde

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Aim: To develop a treatment protocol for the management of tinnitus through multi-modality brain stimulation. Methodology: Present study included 33 adults with unilateral (31 subjects) and bilateral (2 subjects) chronic tinnitus with and/or without hearing loss independent of their etiology. The Treatment protocol included 5 consecutive sessions with follow-up of 6 months. Each session was divided into 3 parts: • Pre-treatment: a) Informed consent b) Pitch and loudness matching. • Treatment: Bimanual paper pen task with tinnitus masking for 30 minutes. • Post-treatment: a) Pitch and loudness matching b) Directive counseling and obtaining feedback. Paper-pen task is to be performed bimanually that included carrying out two different writing activities in different context. The level of difficulty of the activities was increased in successive sessions. Narrowband noise of a frequency same as that of tinnitus was presented at 10 dBSL of tinnitus for 30 minutes simultaneously in the ear with tinnitus. Result: The perception of tinnitus was no longer present in 4 subjects while in remaining subjects it reduced to an intensity that its perception no longer troubled them without causing residual facilitation. In all subjects, the intensity of tinnitus decreased by an extent of 45 dB at an average. However, in few subjects, the intensity of tinnitus also decreased by more than 45 dB. The approach resulted in statistically significant reductions in Tinnitus Functional Index and Tinnitus Handicap Inventory scores. The results correlate with pre and post treatment score of Tinnitus Handicap Inventory that dropped from 90% to 0%. Discussion: Brain mapping(qEEG) Studies report that there is multiple parallel overlapping of neural subnetworks in the non-auditory areas of the brain which exhibits abnormal, constant and spontaneous neural activity involved in the perception of tinnitus with each subnetwork and area reflecting a specific aspect of tinnitus percept. The paper pen task and directive counseling are designed and delivered respectively in a way that is assumed to induce normal, rhythmically constant and premeditated neural activity and mask the abnormal, constant and spontaneous neural activity in the above-mentioned subnetworks and the specific non-auditory area. Counseling was focused on breaking the vicious cycle causing and maintaining the presence of tinnitus. Diverting auditory attention alone is insufficient to reduce the perception of tinnitus. Conscious awareness of tinnitus can be suppressed when individuals engage in cognitively demanding tasks of non-auditory nature as the paper pen task used in the present study. To carry out this task selective, divided, sustained, simultaneous and split attention act cumulatively. Bimanual paper pen task represents a top-down activity which underlies brain’s ability to selectively attend to the bimanual written activity as a relevant stimulus and to ignore tinnitus that is the irrelevant stimuli in the present study. Conclusion: The study suggests that this novel treatment approach is cost effective, time saving and efficient to vanish the tinnitus or to reduce the intensity of tinnitus to a negligible level and thereby eliminating the negative reactions towards tinnitus.

Keywords: multi-modality brain stimulation, neural subnetworks, non-auditory areas, paper-pen task, top-down activity

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Authors: Judy M. Obliosca, Olivia Vest, Sandra Poulos, Kelsi Smith, Tammy Ferguson, Abigail Powers Lott, Alicia K. Smith, Yang Xu, Christopher K. Tison

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Post-Traumatic Stress Disorder (PTSD) is a mental health problem that people may develop after experiencing traumatic events such as combat, natural disasters, and major emotional challenges. Tragically, the number of military personnel with PTSD correlates directly with the number of veterans who attempt suicide, with the highest rate in the Army. Research has shown epigenetic risks in those who are prone to several psychiatric dysfunctions, particularly PTSD. Once initiated in response to trauma, epigenetic alterations in particular, the DNA methylation in the form of 5-methylcytosine (5mC) alters chromatin structure and represses gene expression. Current methods to detect DNA methylation, such as bisulfite-based genomic sequencing techniques, are laborious and have massive analysis workflow while still having high error rates. A faster and simpler detection method of high sensitivity and precision would be useful in a clinical setting to confirm potential PTSD etiologies, prevent other psychiatric disorders, and improve military health. A nano-enhanced Surface Plasmon Resonance imaging (SPRi)-based assay that simultaneously detects site-specific 5mC base (termed as PTSD base) in methylated genes related to PTSD is being developed. The arrays on a sensing chip were first constructed for parallel detection of PTSD bases using synthetic and genomic DNA (gDNA) samples. For the gDNA sample extracted from the whole blood of a PTSD patient, the sample was first digested using specific restriction enzymes, and fragments were denatured to obtain single-stranded methylated target genes (ssDNA). The resulting mixture of ssDNA was then injected into the assay platform, where targets were captured by specific DNA aptamer probes previously immobilized on the surface of a sensing chip. The PTSD bases in targets were detected by anti-5-methylcytosine antibody (anti-5mC), and the resulting signals were then enhanced by the universal nanoenhancer. Preliminary results showed successful detection of a PTSD base in a gDNA sample. Brighter spot images and higher delta values (control-subtracted reflectivity signal) relative to those of the control were observed. We also implemented the in-house surface activation system for detection and developed SPRi disposable chips. Multiplexed PTSD base detection of target methylated genes in blood DNA from PTSD patients of severity conditions (asymptomatic and severe) was conducted. This diagnostic capability being developed is a platform technology, and upon successful implementation for PTSD, it could be reconfigured for the study of a wide variety of neurological disorders such as traumatic brain injury, Alzheimer’s disease, schizophrenia, and Huntington's disease and can be extended to the analyses of other sample matrices such as urine and saliva.

Keywords: epigenetic assay, DNA methylation, PTSD, whole blood, multiplexing

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Authors: Alexander A. Stakheev, Larisa V. Samokhvalova, Sergey K. Zavriev

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Fusarium fungi are among the most devastating plant pathogens distributed all over the world. Significant reduction of grain yield and quality caused by Fusarium leads to multi-billion dollar annual losses to the world agricultural production. These organisms can also cause infections in immunocompromised persons and produce the wide range of mycotoxins, such as trichothecenes, fumonisins, and zearalenone, which are hazardous to human and animal health. Identification of Fusarium fungi based on the morphology of spores and spore-forming structures, colony color and appearance on specific culture media is often very complicated due to the high similarity of these features for closely related species. Modern Fusarium taxonomy increasingly uses data of crossing experiments (biological species concept) and genetic polymorphism analysis (phylogenetic species concept). A number of novel Fusarium sibling species has been established using DNA barcoding techniques. Species recognition is best made with the combined phylogeny of intron-rich protein coding genes and ribosomal DNA sequences. However, the internal transcribed spacer of (ITS), which is considered to be universal DNA barcode for Fungi, is not suitable for genus Fusarium, because of its insufficient variability between closely related species and the presence of non-orthologous copies in the genome. Nowadays, the translation elongation factor 1 alpha (TEF1α) gene is the “gold standard” of Fusarium taxonomy, but the search for novel informative markers is still needed. In this study, we used two novel DNA markers, frataxin (FXN) and heat shock protein 90 (HSP90) to discover phylogenetic relationships between Fusarium species. Multilocus phylogenetic analysis based on partial sequences of TEF1α, FXN, HSP90, as well as intergenic spacer of ribosomal DNA (IGS), beta-tubulin (β-TUB) and phosphate permease (PHO) genes has been conducted for 120 isolates of 19 Fusarium species from different climatic zones of Russia and neighboring countries using maximum likelihood (ML) and maximum parsimony (MP) algorithms. Our analyses revealed that FXN and HSP90 genes could be considered as informative phylogenetic markers, suitable for evolutionary and taxonomic studies of Fusarium genus. It has been shown that PHO gene possesses more variable (22 %) and parsimony informative (19 %) characters than other markers, including TEF1α (12 % and 9 %, correspondingly) when used for elucidating phylogenetic relationships between F. avenaceum and its closest relatives – F. tricinctum, F. acuminatum, F. torulosum. Application of novel DNA barcodes confirmed the fact that F. arthrosporioides do not represent a separate species but only a subspecies of F. avenaceum. Phylogeny based on partial PHO and FXN sequences revealed the presence of separate cluster of four F. avenaceum strains which were closer to F. torulosum than to major F. avenaceum clade. The strain F-846 from Moldova, morphologically identified as F. poae, formed a separate lineage in all the constructed dendrograms, and could potentially be considered as a separate species, but more information is needed to confirm this conclusion. Variable sites in PHO sequences were used for the first-time development of specific qPCR-based diagnostic assays for F. acuminatum and F. torulosum. This work was supported by Russian Foundation for Basic Research (grant № 15-29-02527).

Keywords: DNA barcode, fusarium, identification, phylogenetics, taxonomy

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Authors: Robert Pocklington

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Most current language courses expect students to be ‘vocational’, sacrificing their free time in order to learn. However, pupils with a full-time job, or bringing up children, hardly have a spare moment. Others just need the language as a tool or a qualification, as if it were book-keeping or a driving license. Then there are children in unstructured families whose stressful life makes private study almost impossible. And the countless parents whose evenings and weekends have become a nightmare, trying to get the children to do their homework. There are many arguments against homework being a necessity (rather than an optional extra for more ambitious or dedicated students), making a clear case for teaching methods which facilitate full learning of the key content within the classroom. A methodology which could be described as Programmed Mastery Learning has been used at Fluency Language Academy (Spain) since 1992, to teach English to over 4000 pupils yearly, with a staff of around 100 teachers, barely requiring homework. The course is structured according to the tenets of Programmed Learning: small manageable teaching steps, immediate feedback, and constant successful activity. For the Mastery component (not stopping until everyone has learned), the memorisation and practice are entrusted to flashcard-based drilling in the classroom, leading all students to progress together and develop a permanently growing knowledge base. Vocabulary and expressions are memorised using flashcards as stimuli, obliging the brain to constantly recover words from the long-term memory and converting them into reflex knowledge, before they are deployed in sentence building. The use of grammar rules is practised with ‘cue’ flashcards: the brain refers consciously to the grammar rule each time it produces a phrase until it comes easily. This automation of lexicon and correct grammar use greatly facilitates all other language and conversational activities. The full B2 course consists of 48 units each of which takes a class an average of 17,5 hours to complete, allowing the vast majority of students to reach B2 level in 840 class hours, which is corroborated by an 85% pass-rate in the Cambridge University B2 exam (First Certificate). In the past, studying for qualifications was just one of many different options open to young people. Nowadays, youngsters need to stay at school and obtain qualifications in order to get any kind of job. There are many students in our classes who have little intrinsic interest in what they are studying; they just need the certificate. In these circumstances and with increasing government pressure to minimise failure, teachers can no longer think ‘If they don’t study, and fail, its their problem’. It is now becoming the teacher’s problem. Teachers are ever more in need of methods which make their pupils successful learners; this means assuring learning in the classroom. Furthermore, homework is arguably the main divider between successful middle-class schoolchildren and failing working-class children who drop out: if everything important is learned at school, the latter will have a much better chance, favouring inclusiveness in the language classroom.

Keywords: flashcard drilling, fluency method, mastery learning, programmed learning, teaching English as a foreign language

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Authors: Elena Bellini, Daniele Mugnaini, Michele Boschetto

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People with an Autistic Spectrum Disorder and an Intellectual Disability who need to attend a Hospital Emergency Waiting Room frequently present high levels of discomfort and challenging behaviors due to stress-related hyperarousal, sensory sensitivity, novelty-anxiety, communication and self-regulation difficulties. Increased agitation and acting out also disturb the diagnostic and therapeutic processes, and the emergency room climate. Architectural design disciplines aimed at reducing distress in hospitals or creating autism-friendly environments are called for to find effective answers to this particular need. A growing number of researchers are considering the physical environment as an important point of intervention for people with autism. It has been shown that providing the right setting can help enhance confidence and self-esteem and can have a profound impact on their health and wellbeing. Environmental psychology has evaluated the perceived quality of care, looking at the design of hospital rooms, paths and circulation, waiting rooms, services and devices. Furthermore, many studies have investigated the influence of the hospital environment on patients, in terms of stress-reduction and therapeutic intervention’ speed, but also on health professionals and their work. Several services around the world are organizing autism-friendly hospital environments which involve the architecture and the specific staff training. In Italy, the association Spes contra spem has promoted and published, in 2013, the ‘Chart of disabled people in the hospital’. It stipulates that disabled people should have equal rights to accessible and high-quality care. There are a few Italian examples of therapeutic programmes for autistic people as the Dama project in Milan and the recent experience of Children and Autism Foundation in Pordenone. Careggi’s Emergency Waiting Room in Florence has been built to satisfy this challenge. This project of research comes from a collaboration between the technical staff of Careggi Hospital, the Center for autism PAMAPI and some architects expert in the sensory environment. The methodology of focus group involved architects, psychologists and professionals through a transdisciplinary research, centered on the links between the spatial characteristics and clinical state of people with ASD. The relationship between architectural space and quality of life is studied to pay maximum attention to users’ needs and to support the medical staff in their work by a specific program of training. The result of this research is a sum of criteria used to design the emergency waiting room, that will be illustrated. A protected room, with a clear space design, maximizes comprehension and predictability. The multisensory environment is thought to help sensory integration and relaxation. Visual communication through Ipad allows an anticipated understanding of medical procedures, and a specific technological system supports requests, choices and self-determination in order to fit sensory stimulation to personal preferences, especially for hypo and hypersensitive people. All these characteristics should ensure a better regulation of the arousal, less behavior problems, improving treatment accessibility, safety, and effectiveness. First results about patient-satisfaction levels will be presented.

Keywords: accessibility of care, autism-friendly architecture, personalized therapeutic process, sensory environment

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Authors: Wenyu Guo, Malachy O’Rourke, Mark Bowkett, Michael Gilchrist

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Many devices for real-time monitoring of surface water have been developed in the past few years to provide early warning of pollutions and so to decrease the risk of environmental pollution efficiently. One of the most common methodologies used in the detection system is a colorimetric process, in which a container with fixed volume is filled with target ions and reagents to combine a colorimetric dye. The colorimetric ions can sensitively absorb a specific-wavelength radiation beam, and its absorbance rate is proportional to the concentration of the fully developed product, indicating the concentration of target nutrients in the pre-mixed water samples. In order to achieve precise and rapid detection effect, channels with dimensions in the order of micrometers, i.e., microfluidic systems have been developed and introduced into these diagnostics studies. Microfluidics technology largely reduces the surface to volume ratios and decrease the samples/reagents consumption significantly. However, species transport in such miniaturized channels is limited by the low Reynolds numbers in the regimes. Thus, the flow is extremely laminar state, and diffusion is the dominant mass transport process all over the regimes of the microfluidic channels. The objective of this present work has been to analyse the mixing effect and chemistry kinetics in a stop-flow microfluidic device measuring Nitride concentrations in fresh water samples. In order to improve the temporal resolution of the Nitride microfluidic sensor, we have used computational fluid dynamics to investigate the influence that the effectiveness of the mixing process between the sample and reagent within a microfluidic device exerts on the time to completion of the resulting chemical reaction. This computational approach has been complemented by physical experiments. The kinetics of the Griess reaction involving the conversion of sulphanilic acid to a diazonium salt by reaction with nitrite in acidic solution is set in the Laminar Finite-rate chemical reaction in the model. Initially, a methodology was developed to assess the degree of mixing of the sample and reagent within the device. This enabled different designs of the mixing channel to be compared, such as straight, square wave and serpentine geometries. Thereafter, the time to completion of the Griess reaction within a straight mixing channel device was modeled and the reaction time validated with experimental data. Further simulations have been done to compare the reaction time to effective mixing within straight, square wave and serpentine geometries. Results show that square wave channels can significantly improve the mixing effect and provides a low standard deviations of the concentrations of nitride and reagent, while for straight channel microfluidic patterns the corresponding values are 2-3 orders of magnitude greater, and consequently are less efficiently mixed. This has allowed us to design novel channel patterns of micro-mixers with more effective mixing that can be used to detect and monitor levels of nutrients present in water samples, in particular, Nitride. Future generations of water quality monitoring and diagnostic devices will easily exploit this technology.

Keywords: nitride detection, computational fluid dynamics, chemical kinetics, mixing effect

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Authors: Molly Parker

Abstract:

Unintended pregnancy rates in Australia are amongst the highest in the developed world. Women with Substance Use Disorder often have riskier sexual behavior with nil contraceptive use and face disproportionately higher unintended pregnancies and Sexually Transmitted Infections, alongside Substance Use in Pregnancy (SUP) climbing at an alarming rate. In an inner-city Drug and Alcohol (D&A) service, significant barriers to sexual and reproductive health services have been identified, aligning with research. Rapid pathways were created for women seeking D&A treatment to be referred to Sexual and Reproductive Health services for the administration of Long-acting reversible contraception (LARC) and sexual health screening. For clients attending a D&A service, this is an opportunistic time to offer sexual and reproductive health services. Collaboration and multidisciplinary team input between D&A and sexual health and reproductive services are paramount, with rapid referral pathways being identified as the main strategy to improve access to sexual and reproductive health support for this population. With this evidence, a rapid referral pathway was created for women using the D&A service to access LARC, particularly in view of fertility often returning once stable on D&A treatment. A closed-ended survey was used for D&A staff to identify gaps in reproductive health knowledge and views of referral accessibility. Results demonstrated a lack of knowledge of contraception and appropriate referral processes. A closed-ended survey for clients was created to establish the need and access to services and to quantify data. A follow-up data collection will be reviewed to access uptake and satisfaction of the intervention from clients. Sexual health screening access was also identified as a deficit, particularly concerning due to the higher rates of STIs in this cohort. A rapid referral pathway will be undergoing implementation, reducing risks of untreated STIS both pre and post-conception. Similarly, pre and post-intervention structured surveys will be used to identify client satisfaction from the pathway. Although currently in progress, the research and pathway aim to be completed by December 2023. This research and implementation of sexual and reproductive health pathways from the D&A service have significant health and well-being benefits to clients and the wider community, including possible fetal/infancy outcomes. Women now have rapid access to sexual and reproductive health services, with the aim of reducing unplanned pregnancies, poor outcomes associated with SUP, client/staff trauma from termination of pregnancy, and client/staff trauma following the assumption of care of the child due to substance use, the financial cost for out of home care as required, the poor outcomes of untreated STIs to the fetus in pregnancy and the spread of STIs in the wider community. As evidence suggests, the implementation of a streamlined referral process is required between D&A and sexual and reproductive health services and has positive feedback from both clinicians and clients in improving care.

Keywords: substance use in pregnancy, drug and alcohol, substance use disorder, sexual health, reproductive health, contraception, long-acting reversible contraception, neonatal abstinence syndrome, FASD, sexually transmitted infections, sexually transmitted infections pregnancy

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Authors: Sharon Harel, Rachel Kizony, Yoram Feldman, Gabi Zeilig, Mordechai Shani

Abstract:

Multiple Sclerosis (MS) is a neurological disease that may cause restriction in participation in daily activities of young adults. Main symptoms include fatigue, weakness and cognitive decline. The appearance of symptoms, their severity and deterioration rate, change between patients. The challenge of health services is to provide long-term rehabilitation services to people with MS. The objective of this presentation is to describe a course of tele-rehabilitation service of a woman with MS. Methods; R is a 48 years-old woman, diagnosed with MS when she was 22. She started to suffer from weakness of her non-dominant left upper extremity about ten years after the diagnosis. She was referred to the tele-rehabilitation service by her rehabilitation team, 16 years after diagnosis. Her goals were to improve ability to use her affected upper extremity in daily activities. On admission her score in the Mini-Mental State Exam was 30/30. Her Fugl-Meyer Assessment (FMA) score of the left upper extremity was 48/60, indicating mild weakness and she had a limitation of her shoulder abduction (90 degrees). In addition, she reported little use of her arm in daily activities as shown in her responses to the Motor Activity Log (MAL) that were equal to 1.25/5 in amount and 1.37 in quality of use. R. received two 30 minutes on-line sessions per week in the tele-rehabilitation service, with the CogniMotion system. These were complemented by self-practice with the system. The CogniMotion system provides a hybrid (synchronous-asynchronous), the home-based tele-rehabilitation program to improve the motor, cognitive and functional status of people with neurological deficits. The system consists of a computer, large monitor, and the Microsoft’s Kinect 3D sensor. This equipment is located in the client’s home and connected to a clinician’s computer setup in a remote clinic via WiFi. The client sits in front of the monitor and uses his body movements to interact with games and tasks presented on the monitor. The system provides feedback in the form of ‘knowledge of results’ (e.g., the success of a game) and ‘knowledge of performance’ (e.g., alerts for compensatory movements) to enhance motor learning. The games and tasks were adapted for R. motor abilities and level of difficulty was gradually increased according to her abilities. The results of her second assessment (after 35 on-line sessions) showed improvement in her FMA score to 52 and shoulder abduction to 140 degrees. Moreover, her responses to the MAL indicated an increased amount (2.4) and quality (2.2) of use of her left upper extremity in daily activities. She reported high level of enjoyment from the treatments (5/5), specifically the combination of cognitive challenges while moving her body. In addition, she found the system easy to use as reflected by her responses to the System Usability Scale (85/100). To-date, R. continues to receive treatments in the tele-rehabilitation service. To conclude, this case report shows the potential of using tele-rehabilitation for people with MS to provide strategies to enhance the use of the upper extremity in daily activities as well as for maintaining motor function.

Keywords: motor function, multiple-sclerosis, tele-rehabilitation, daily activities

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Authors: Priyanka Jain, Ashish K. Sharma, Esha Shukla, K. J. Mukherjee

Abstract:

We propose a paradigm shift in our approach to design improved platforms for recombinant protein production, by addressing system level issues rather than the individual steps associated with recombinant protein synthesis like transcription, translation, etc. We demonstrate that by controlling and modulating the cellular stress response (CSR), which is responsible for feedback control of protein synthesis, we can generate hyper-producing strains. We did transcriptomic profiling of post-induction cultures, expressing different types of protein, to analyze the nature of this cellular stress response. We found significant down-regulation of substrate utilization, translation, and energy metabolism genes due to generation CSR inside the host cell. However, transcription profiling has also shown that many genes are up-regulated post induction and their role in modulating the CSR is unclear. We hypothesized that these up-regulated genes trigger signaling pathways, generating the CSR and concomitantly reduce the recombinant protein yield. To test this hypothesis, we knocked out the up-regulated genes, which did not have any downstream regulatees, and analyzed their impact on cellular health and recombinant protein expression. Two model proteins i.e., GFP and L-Asparaginase were chosen for this analysis. We observed a significant improvement in expression levels, with some knock-outs showing more than 7-fold higher expression compared to control. The 10 best single knock-outs were chosen to make 45 combinations of all possible double knock-outs. A further increase in expression was observed in some of these double knock- outs with GFP levels being highest in a double knock-out ΔyhbC + ΔelaA. However, for L-Asparaginase which is a secretory protein, the best results were obtained using a combination of ΔelaA+ΔcysW knock-outs. We then tested all the knock outs for their ability to enhance the expression of a 'difficult-to-express' protein. The Rubella virus E1 protein was chosen and tagged with sfGFP at the C-terminal using a linker peptide for easy online monitoring of expression of this fusion protein. Interestingly, the highest increase in Rubella-sGFP levels was obtained in the same double knock-out ΔelaA + ΔcysW (5.6 fold increase in expression yield compared to the control) which gave the highest expression for L-Asparaginase. However, for sfGFP alone, the ΔyhbC+ΔmarR knock-out gave the highest level of expression. These results indicate that there is a fair degree of commonality in the nature of the CSR generated by the induction of different proteins. Transcriptomic profiling of the double knock out showed that many genes associated with the translational machinery and energy biosynthesis did not get down-regulated post induction, unlike the control where these genes were significantly down-regulated. This confirmed our hypothesis of these genes playing an important role in the generation of the CSR and allowed us to design a strategy for making better expression hosts by simply knocking out key genes. This strategy is radically superior to the previous approach of individually up-regulating critical genes since it blocks the mounting of the CSR thus preventing the down-regulation of a very large number of genes responsible for sustaining the flux through the recombinant protein production pathway.

Keywords: cellular stress response, GFP, knock-outs, up-regulated genes

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Authors: Patrick Cormier, Cesar Meylan, Matt Jensen, Dana Agar-Newman, Chloe Werle, Ming-Chang Tsai, Marc Klimstra

Abstract:

Sprint-derived horizontal force-velocity-power (FVP) profiles can be developed with adequate validity and reliability with satellite (GPS/GNSS) systems. However, FVP metrics are sensitive to small nuances in data processing procedures such that minor differences in defining the onset and end of the sprint could result in different FVP metric outcomes. Furthermore, in team-sports, there is a requirement for rapid analysis and feedback of results from multiple athletes, therefore developing standardized and automated methods to improve the speed, efficiency and reliability of this process are warranted. Thus, the purpose of this study was to compare different methods of sprint end detection on the development of FVP profiles from 10Hz GPS/GNSS data through goodness-of-fit and intertrial reliability statistics. Seventeen national team female soccer players participated in the FVP protocol which consisted of 2x40m maximal sprints performed towards the end of a soccer specific warm-up in a training session (1020 hPa, wind = 0, temperature = 30°C) on an open grass field. Each player wore a 10Hz Catapult system unit (Vector S7, Catapult Innovations) inserted in a vest in a pouch between the scapulae. All data were analyzed following common procedures. Variables computed and assessed were the model parameters, estimated maximal sprint speed (MSS) and the acceleration constant τ, in addition to horizontal relative force (F₀), velocity at zero (V₀), and relative mechanical power (Pmax). The onset of the sprints was standardized with an acceleration threshold of 0.1 m/s². The sprint end detection methods were: 1. Time when peak velocity (MSS) was achieved (zero acceleration), 2. Time after peak velocity drops by -0.4 m/s, 3. Time after peak velocity drops by -0.6 m/s, and 4. When the integrated distance from the GPS/GNSS signal achieves 40-m. Goodness-of-fit of each sprint end detection method was determined using the residual sum of squares (RSS) to demonstrate the error of the FVP modeling with the sprint data from the GPS/GNSS system. Inter-trial reliability (from 2 trials) was assessed utilizing intraclass correlation coefficients (ICC). For goodness-of-fit results, the end detection technique that used the time when peak velocity was achieved (zero acceleration) had the lowest RSS values, followed by -0.4 and -0.6 velocity decay, and 40-m end had the highest RSS values. For intertrial reliability, the end of sprint detection techniques that were defined as the time at (method 1) or shortly after (method 2 and 3) when MSS was achieved had very large to near perfect ICC and the time at the 40 m integrated distance (method 4) had large to very large ICCs. Peak velocity was reached at 29.52 ± 4.02-m. Therefore, sport scientists should implement end of sprint detection either when peak velocity is determined or shortly after to improve goodness of fit to achieve reliable between trial FVP profile metrics. Although, more robust processing and modeling procedures should be developed in future research to improve sprint model fitting. This protocol was seamlessly integrated into the usual training which shows promise for sprint monitoring in the field with this technology.

Keywords: automated, biomechanics, team-sports, sprint

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Authors: Andy Alubaidy

Abstract:

In this work, a robotic additive manufacturing system (RAMS) that is capable of three-dimensional (3D) printing in six degrees of freedom (DOF) with very high accuracy and virtually on any surface has been designed and built. One of the major shortcomings in existing 3D printer technology is the limitation to three DOF, which results in prolonged fabrication time. Depending on the techniques used, it usually takes at least two hours to print small objects and several hours for larger objects. Another drawback is the size of the printed objects, which is constrained by the physical dimensions of most low-cost 3D printers, which are typically small. In such cases, large objects are produced by dividing them into smaller components that fit the printer’s workable area. They are then glued, bonded or otherwise attached to create the required object. Another shortcoming is material constraints and the need to fabricate a single part using different materials. With the flexibility of a six-DOF robot, the RAMS has been designed to overcome these problems. A feeding mechanism using an adaptive Proportional-Integral-Derivative (PID) controller is utilized along with a national instrument compactRIO (NI cRIO), an ABB robot, and off-the-shelf sensors. The RAMS have the ability to 3D print virtually anywhere in six degrees of freedom with very high accuracy. It is equipped with an ABB IRB 120 robot to achieve this level of accuracy. In order to convert computer-aided design (CAD) files to digital format that is acceptable to the robot, Hypertherm Robotic Software Inc.’s state-of-the-art slicing software called “ADDMAN” is used. ADDMAN is capable of converting any CAD file into RAPID code (the programing language for ABB robots). The robot uses the generated code to perform the 3D printing. To control the entire process, National Instrument (NI) compactRIO (cRio 9074), is connected and communicated with the robot and a feeding mechanism that is designed and fabricated. The feeding mechanism consists of two major parts, cold-end and hot-end. The cold-end consists of what is conventionally known as an extruder. Typically, a stepper-motor is used to control the push on the material, however, for optimum control, a DC motor is used instead. The hot-end consists of a melt-zone, nozzle, and heat-brake. The melt zone ensures a thorough melting effect and consistent output from the nozzle. Nozzles are made of brass for thermo-conductivity while the melt-zone is comprised of a heating block and a ceramic heating cartridge to transfer heat to the block. The heat-brake ensures that there is no heat creep-up effect as this would swell the material and prevent consistent extrusion. A control system embedded in the cRio is developed using NI Labview which utilizes adaptive PID to govern the heating cartridge in conjunction with a thermistor. The thermistor sends temperature feedback to the cRio, which will issue heat increase or decrease based on the system output. Since different materials have different melting points, our system will allow us to adjust the temperature and vary the material.

Keywords: robotic, additive manufacturing, PID controller, cRIO, 3D printing

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Authors: Arifur Rahman, Ardeshir Amirkhani, Honghua Hu, Mark Molloy, Karen Vickery

Abstract:

Introduction and Objectives: Staphylococcus aureus and coagulase-negative staphylococci comprises approximately 65% of infections associated with medical devices and are well known for their biofilm formatting ability. Biofilm-related infections are extremely difficult to eradicate owing to their high tolerance to antibiotics and host immune defences. Currently, there is no efficient method for early biofilm detection. A better understanding to enable detection of biofilm specific proteins in vitro and in vivo can be achieved by studying planktonic and different growth phases of biofilms using a proteome analysis approach. Our goal was to construct a reference map of planktonic and biofilm associated proteins of S. aureus. Methods: S. aureus reference strain (ATCC 25923) was used to grow 24 hours planktonic, 3-day wet biofilm (3DWB), and 12-day wet biofilm (12DWB). Bacteria were grown in tryptic soy broth (TSB) liquid medium. Planktonic growth was used late logarithmic bacteria, and the Centres for Disease Control (CDC) biofilm reactor was used to grow 3 days, and 12-day hydrated biofilms, respectively. Samples were subjected to reduction, alkylation and digestion steps prior to Multiplex labelling using Tandem Mass Tag (TMT) 10-plex reagent (Thermo Fisher Scientific). The labelled samples were pooled and fractionated by high pH RP-HPLC which followed by loading of the fractions on a nanoflow UPLC system (Eksigent UPLC system, AB SCIEX). Mass spectrometry (MS) data were collected on an Orbitrap Elite (Thermo Fisher Scientific) Mass Spectrometer. Protein identification and relative quantitation of protein levels were performed using Proteome Discoverer (version 1.3, Thermo Fisher Scientific). After the extraction of protein ratios with Proteome Discoverer, additional processing, and statistical analysis was done using the TMTPrePro R package. Results and Discussion: The present study showed that a considerable proteomic difference exists among planktonic and biofilms from S. aureus. We identified 1636 total extracellular secreted proteins, of which 350 and 137 proteins of 3DWB and 12DWB showed significant abundance variation from planktonic preparation, respectively. Of these, simultaneous up-regulation in between 3DWB and 12DWB proteins such as extracellular matrix-binding protein ebh, enolase, transketolase, triosephosphate isomerase, chaperonin, peptidase, pyruvate kinase, hydrolase, aminotransferase, ribosomal protein, acetyl-CoA acetyltransferase, DNA gyrase subunit A, glycine glycyltransferase and others we found in this biofilm producer. On the contrary, simultaneous down-regulation in between 3DWB and 12DWB proteins such as alpha and delta-hemolysin, lipoteichoic acid synthase, enterotoxin I, serine protease, lipase, clumping factor B, regulatory protein Spx, phosphoglucomutase, and others also we found in this biofilm producer. In addition, we also identified a big percentage of hypothetical proteins including unique proteins. Therefore, a comprehensive knowledge of planktonic and biofilm associated proteins identified by S. aureus will provide a basis for future studies on the development of vaccines and diagnostic biomarkers. Conclusions: In this study, we constructed an initial reference map of planktonic and various growth phase of biofilm associated proteins which might be helpful to diagnose biofilm associated infections.

Keywords: bacterial biofilms, CDC bioreactor, S. aureus, mass spectrometry, TMT

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