Search results for: multidimensional functions

Authors: Rena Upitis, Philip C. Abrami, Karen Boese

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The success of one-on-one music study relies heavily on the ability of the teacher to provide sufficient direction to students during weekly lessons so that they can successfully practice from one lesson to the next. Traditionally, these instructions are given in a paper notebook, where the teacher makes notes for the students after describing a task or demonstrating a technique. The ability of students to make sense of these notes varies according to their understanding of the teacher’s directions, their motivation to practice, their memory of the lesson, and their abilities to self-regulate. At best, the notes enable the student to progress successfully. At worst, the student is left rudderless until the next lesson takes place. Digital notebooks have the potential to provide a more interactive and effective bridge between music lessons than traditional pen-and-paper notebooks. One such digital notebook, Cadenza, was designed to streamline and improve teachers’ instruction, to enhance student practicing, and to provide the means for teachers and students to communicate between lessons. For example, Cadenza contains a video annotator, where teachers can offer real-time guidance on uploaded student performances. Using the checklist feature, teachers and students negotiate the frequency and type of practice during the lesson, which the student can then access during subsequent practice sessions. Following the tenets of self-regulated learning, goal setting and reflection are also featured. Accordingly, the present paper addressed the following research questions: (1) How does the use of the Cadenza digital music notebook engage students and their teachers?, (2) Which features of Cadenza are most successful?, (3) Which features could be improved?, and (4) Is student learning and motivation enhanced with the use of the Cadenza digital music notebook? The paper describes the results 10 months of field-testing of Cadenza, structured around the four research questions outlined. Six teachers and 65 students took part in the study. Data were collected through video-recorded lesson observations, digital screen captures, surveys, and interviews. Standard qualitative protocols for coding results and identifying themes were employed to analyze the results. The results consistently indicated that teachers and students embraced the digital platform offered by Cadenza. The practice log and timer, the real-time annotation tool, the checklists, the lesson summaries, and the commenting features were found to be the most valuable functions, by students and teachers alike. Teachers also reported that students progressed more quickly with Cadenza, and received higher results in examinations than those students who were not using Cadenza. Teachers identified modifications to Cadenza that would make it an even more powerful way to support student learning. These modifications, once implemented, will move the tool well past its traditional notebook uses to new ways of motivating students to practise between lessons and to communicate with teachers about their learning. Improvements to the tool called for by the teachers included the ability to duplicate archived lessons, allowing for split screen viewing, and adding goal setting to the teacher window. In the concluding section, proposed modifications and their implications for self-regulated learning are discussed.

Keywords: digital music technologies, electronic notebooks, self-regulated learning, studio music instruction

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Authors: Sumeet Rai, Anuj Tyagi, B. T. Naveen Kumar, Shubhkaramjeet Kaur, Niraj K. Singh

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Since their discovery, indiscriminate use of antibiotics in human, veterinary and aquaculture systems has resulted in global emergence/spread of multidrug-resistant bacterial pathogens. Thus, the need for alternative approaches to control bacterial infections has become utmost important. High selectivity/specificity of bacteriophages (phages) permits the targeting of specific bacteria without affecting the desirable flora. In this study, a lytic phage (Ahp1) specific to Aeromonas hydrophila subsp. hydrophila was isolated from finfish aquaculture pond. The host range of Ahp1 range was tested against 10 isolates of A. hydrophila, 7 isolates of A. veronii, 25 Vibrio cholerae isolates, 4 V. parahaemolyticus isolates and one isolate each of V. harveyi and Salmonella enterica collected previously. Except the host A. hydrophila subsp. hydrophila strain, no lytic activity against any other bacterial was detected. During the adsorption rate and one-step growth curve analysis, 69.7% of phage particles were able to get adsorbed on host cell followed by the release of 93 ± 6 phage progenies per host cell after a latent period of ~30 min. Phage nucleic acid was extracted by column purification methods. After determining the nature of phage nucleic acid as dsDNA, phage genome was subjected to next-generation sequencing by generating paired-end (PE, 2 x 300bp) reads on Illumina MiSeq system. De novo assembly of sequencing reads generated circular phage genome of 42,439 bp with G+C content of 58.95%. During open read frame (ORF) prediction and annotation, 22 ORFs (out of 49 total predicted ORFs) were functionally annotated and rest encoded for hypothetical proteins. Proteins involved in major functions such as phage structure formation and packaging, DNA replication and repair, DNA transcription and host cell lysis were encoded by the phage genome. The complete genome sequence of Ahp1 along with gene annotation was submitted to NCBI GenBank (accession number MF683623). Stability of Ahp1 preparations at storage temperatures of 4 °C, 30 °C, and 40 °C was studied over a period of 9 months. At 40 °C storage, phage counts declined by 4 log units within one month; with a total loss of viability after 2 months. At 30 °C temperature, phage preparation was stable for < 5 months. On the other hand, phage counts decreased by only 2 log units over a period of 9 during storage at 4 °C. As some of the phages have also been reported as glycerol sensitive, the stability of Ahp1 preparations in (0%, 15%, 30% and 45%) glycerol stocks were also studied during storage at -80 °C over a period of 9 months. The phage counts decreased only by 2 log units during storage, and no significant difference in phage counts was observed at different concentrations of glycerol. The Ahp1 phage discovered in our study had a very narrow host range and it may be useful for phage typing applications. Moreover, the endolysin and holin genes in Ahp1 genome could be ideal candidates for recombinant cloning and expression of antimicrobial proteins.

Keywords: Aeromonas hydrophila, endolysin, phage, narrow host range

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Authors: Noam Markovitz

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People with physical disabilities constitute a very large and simultaneously a diverse group of general population, as the term physical disabilities is extensive and covers a wide range of disabilities. Therefore, individuals with physical disabilities are often faced with a new, threatening and stressful reality leading possibly to a multi-crisis in their lives due to the great changes they experience in somatic, socio-economic, occupational and psychological level. The current study seeks to advance understanding of the complex adaptation to physical disabilities by expanding the dynamic-adaptational model of the pursuit of happiness in a hostile world with a new conception of physical self-integrity. Physical self-integrity incorporates an objective dimension, namely physical self-functioning (PSF), and a subjective dimension, namely physical self-concept (PSC). Both of these dimensions constitute an experience of wholeness in the individual’s identification with her or his physical body. The model guiding this work is dialectical in nature and depicts two systems in the individual’s sense of happiness: subjective well-being (SWB) and meaning in life (MIL). Both systems serve as self-adaptive agents that moderate the complementary system of the hostile-world scenario (HWS), which integrates one’s perceived threats to one’s integrity. Thus, in situations of increased HWS, the moderation may take a form of joint activity in which SWB and MIL are amplified or a form of compensation in which one system produces a stronger effect while the other system produces a weaker effect. The current study investigated PSC in relations to SWB and MIL through pleasantness and meanings that are physically or metaphorically grounded in one’s body. In parallel, PSC also relates to HWS by activating representations of inappropriateness, deformation and vulnerability. In view of possibly dialectical positions of opposing and complementary forces within the current model, the current field study that aims to explore PSC as appearing in an independent, cross-sectional, design addressing the model’s variables in a focal group of people with physical disabilities. This study delineated the participation of the PSC in the adaptational functions of SWB and MIL vis-à-vis HWS-related life adversities. The findings showed that PSC could fully complement the main variables of the pursuit of happiness in a hostile world model. The assumed dialectics in the form of a stronger relationship between SWB and MIL in the face of physical disabilities was not supported. However, it was found that when HWS increased, PSC and MIL were strongly linked, whereas PSC and SWB were weakly linked. This highlights the compensatory role of MIL. From a conceptual viewpoint, the current investigation may clarify the role of PSC as an adaptational agent of the individual’s positive health in complementary senses of bodily wholeness. Methodologically, the advantage of the current investigation is the application of an integrative, model-based approach within a specially focused design with a particular relevance to PSC. Moreover, from an applicative viewpoint, the current investigation may suggest how an innovative model may be translated to therapeutic interventions used by clinicians, counselors and practitioners in improving wellness and psychological well-being, particularly among people with physical disabilities.

Keywords: older adults, physical disabilities, physical self-concept, pursuit of happiness in a hostile-world

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Authors: Aleksandr S. Demin, Anastasiia V. Ivanova

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Purpose: The current economy tends to increase the influence of digital technologies and diminish the human role in management. However, it is impossible to deny that a person still leads a business with its own set of values and priorities. The article presented aims to incorporate the peculiarities of the national culture and the characteristics of the supply chain using the quantitative values of the national culture obtained by the scholars of comparative management (Hofstede, House, and others). Design/Methodology/Approach: The conducted research is based on the secondary data in the field of cross-country comparison achieved by Prof. Hofstede and received in the GLOBE project. The data mentioned are used to design different aspects of the supply chain both on the cross-functional and inter-organizational levels. The connection between a range of principles in general (roles assignment, customer service prioritization, coordination of supply chain partners) and in comparative management (acknowledgment of the national peculiarities of the country in which the company operates) is shown over economic and mathematical models, mainly linear programming models. Findings: The combination of the team management wheel concept, the business processes of the global supply chain, and the national culture characteristics let a transnational corporation to form a supply chain crew balanced in costs, functions, and personality. To elaborate on an effective customer service policy and logistics strategy in goods and services distribution in the country under review, two approaches are offered. The first approach relies exceptionally on the customer’s interest in the place of operation, while the second one takes into account the position of the transnational corporation and its previous experience in order to accord both organizational and national cultures. The effect of integration practice on the achievement of a specific supply chain goal in a specific location is advised to assess via types of correlation (positive, negative, non) and the value of national culture indices. Research Limitations: The models developed are intended to be used by transnational companies and business forms located in several nationally different areas. Some of the inputs to illustrate the application of the methods offered are simulated. That is why the numerical measurements should be used with caution. Practical Implications: The research can be of great interest for the supply chain managers who are responsible for the engineering of global supply chains in a transnational corporation and the further activities in doing business on the international area. As well, the methods, tools, and approaches suggested can be used by top managers searching for new ways of competitiveness and can be suitable for all staff members who are keen on the national culture traits topic. Originality/Value: The elaborated methods of decision-making with regard to the national environment suggest the mathematical and economic base to find a comprehensive solution.

Keywords: logistics integration, logistics services, multinational corporation, national culture, team management, service policy, supply chain management

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Authors: Shubhangi Roy

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Relational Contracts (informal understandings not enforceable by law) are a common feature of most economies. However, their dominance is higher in developing countries. Such informality of economic sectors is often co-related to lower economic growth. The aim of this paper is to investigate whether informal arrangements i.e. relational contracts are a cause or symptom of lower levels of economic and/or institutional development. The methodology followed involves an initial survey of 150 test subjects in Northern India. The subjects are all members of occupations where they frequently transact ensuring uniformity in transaction volume. However, the subjects are from varied socio-economic backgrounds to ensure sufficient variance in transaction values allowing us to understand the relationship between the amount of money involved to the method of transaction used, if any. Questions asked are quantitative and qualitative with an aim to observe both the behavior and motivation behind such behavior. An overarching similarity observed during the survey across all subjects’ responses is that in an economy like India with pervasive corruption and delayed litigation, economy participants have created alternative social sanctions to deal with non-performers. In a society that functions predominantly on caste, class and gender classifications, these sanctions could, in fact, be more cumbersome for a potential rule-breaker than the legal ramifications. It, therefore, is a symptom of weak formal regulatory enforcement and dispute settlement mechanism. Additionally, the study bifurcates such informal arrangements into two separate systems - a) when it exists in addition to and augments a legal framework creating an efficient socio-legal equilibrium or; b) in conflict with the legal system in place. This categorization is an important step in regulating informal arrangements. Instead of considering the entire gamut of such arrangements as counter-development, it helps decision-makers understand when to dismantle (latter) and when to pivot around existing informal systems (former). The paper hypothesizes that those social arrangements that support the formal legal frameworks allow for cheaper enforcement of regulations with lower enforcement costs burden on the state mechanism. On the other hand, norms which contradict legal rules will undermine the formal framework. Law infringement, in presence of these norms, will have no impact on the reputation of the business or individual outside of the punishment imposed under the law. It is especially exacerbated in the Indian legal system where enforcement of penalties for non-performance of contracts is low. In such a situation, the social norm will be adhered to more strictly by the individuals rather than the legal norms. This greatly undermines the role of regulations. The paper concludes with recommendations that allow policy-makers and legal systems to encourage the former category of informal arrangements while discouraging norms that undermine legitimate policy objectives. Through this investigation, we will be able to expand our understanding of tools of market development beyond regulations. This will allow academics and policymakers to harness social norms for less disruptive and more lasting growth.

Keywords: distribution of income, emerging economies, relational contracts, sample survey, social norms

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Authors: K. Premakumar, D. S. Rushani, H. N. Hettiarachchi

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The production and consumption of preserved foods have gained much importance due to globalization, and they provide a health benefit apart from the basic nutritional functions. Therefore, a study was conducted to develop a jam incorporating carrot, banana, and lime. Considering the findings of several preliminary studies, five formulations of the jam were prepared by blending different percentages of carrot and banana including control (where the only carrot was added). The freshly prepared formulations were subjected to physicochemical and sensory analysis.Physico-Chemical parameters such as pH, TSS, titrable acidity, ascorbic acid content, total sugar and non-reducing sugar and organoleptic qualities such as colour, aroma, taste, spread ability and overall acceptability and microbial analysis (total plate count) were analyzed after formulations. Physico-Chemical Analysis of the freshly prepared Carrot –Banana Blend jam showed increasing trend in titrable acidity (from 0.8 to 0.96, as % of citric acid), TSS (from 70.05 to 67.5 0Brix), ascorbic acid content (from 0.83 to 11.465 mg/100ml), reducing sugar (from 15.64 to 20.553%) with increase in carrot pulp from 50 to 100%. pH, total sugar, and non-reducing sugar were also reduced when carrot concentration is increased. Five points hedonic scale was used to evaluate the organoleptic characters. According to Duncan's Multiple Range Test, the mean scores for all the assessed sensory characters varied significantly (p<0.05) in the freshly made carrot-banana blend jam formulations. Based on the physicochemical and sensory analysis, the most preferred carrot: banana combinations of 50:50, 100:0 and 80:20 (T1, T2, and T5) were selected for storage studies.The formulations were stored at 300 °C room temperature and 70-75% of RH for 12 weeks. The physicochemical characteristics were measured at two weeks interval during storage. The decreasing trends in pH and ascorbic acid and an increasing trend in TSS, titrable acidity, total sugar, reducing sugar and non-reducing sugar were noted with advancement of storage periods of 12 weeks. The results of the chemical analysis showed that there were significance differences (p<0.05) between the tested formulations. Sensory evaluation was done for carrot –banana blends jam after a period of 12 weeks through a panel of 16 semi-trained panelists. The sensory analysis showed that there were significant differences (p<0.05) for organoleptic characters between carrot-banana blend jam formulations. The highest overall acceptability was observed in formulation with 80% carrot and 20% banana pulp. Microbiological Analysis was carried out on the day of preparation, 1 month, 2 months and 3 months after preparation. No bacterial growth was observed in the freshly made carrot -banana blend jam. There were no counts of yeast and moulds and coliforms in all treatments after the heat treatments and during the storage period. Only the bacterial counts (Total Plate Counts) were observed after three months of storage below the critical level, and all formulations were microbiologically safe for consumption. Based on the results of physio-chemical characteristics, sensory attributes, and microbial test, the carrot –banana blend jam with 80% carrot and 20% banana (T2) was selected as best formulation and could be stored up to 12 weeks without any significant changes in the quality characteristics.

Keywords: formulations, physicochemical parameters, microbiological analysis, sensory evaluation

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Authors: Ayon Mukherjee

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Covariate-adjusted response-adaptive (CARA) designs use the available responses to skew the treatment allocation in a clinical trial in towards treatment found at an interim stage to be best for a given patient's covariate profile. Extensive research has been done on various aspects of CARA designs with the patient responses assumed to follow a parametric model. However, ranges of application for such designs are limited in real-life clinical trials where the responses infrequently fit a certain parametric form. On the other hand, robust estimates for the covariate-adjusted treatment effects are obtained from the parametric assumption. To balance these two requirements, designs are developed which are free from distributional assumptions about the survival responses, relying only on the assumption of proportional hazards for the two treatment arms. The proposed designs are developed by deriving two types of optimum allocation designs, and also by using a distribution function to link the past allocation, covariate and response histories to the present allocation. The optimal designs are based on biased coin procedures, with a bias towards the better treatment arm. These are the doubly-adaptive biased coin design (DBCD) and the efficient randomized adaptive design (ERADE). The treatment allocation proportions for these designs converge to the expected target values, which are functions of the Cox regression coefficients that are estimated sequentially. These expected target values are derived based on constrained optimization problems and are updated as information accrues with sequential arrival of patients. The design based on the link function is derived using the distribution function of a probit model whose parameters are adjusted based on the covariate profile of the incoming patient. To apply such designs, the treatment allocation probabilities are sequentially modified based on the treatment allocation history, response history, previous patients’ covariates and also the covariates of the incoming patient. Given these information, an expression is obtained for the conditional probability of a patient allocation to a treatment arm. Based on simulation studies, it is found that the ERADE is preferable to the DBCD when the main aim is to minimize the variance of the observed allocation proportion and to maximize the power of the Wald test for a treatment difference. However, the former procedure being discrete tends to be slower in converging towards the expected target allocation proportion. The link function based design achieves the highest skewness of patient allocation to the best treatment arm and thus ethically is the best design. Other comparative merits of the proposed designs have been highlighted and their preferred areas of application are discussed. It is concluded that the proposed CARA designs can be considered as suitable alternatives to the traditional balanced randomization designs in survival trials in terms of the power of the Wald test, provided that response data are available during the recruitment phase of the trial to enable adaptations to the designs. Moreover, the proposed designs enable more patients to get treated with the better treatment during the trial thus making the designs more ethically attractive to the patients. An existing clinical trial has been redesigned using these methods.

Keywords: censored response, Cox regression, efficiency, ethics, optimal allocation, power, variability

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Authors: Andrew N. Saylor, James R. Peters

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Scoliosis is a complex 3D deformity of the thoracic and lumbar spines, clinically diagnosed by measurement of a Cobb angle of 10 degrees or more on a coronal X-ray. The Cobb angle is the angle made by the lines drawn along the proximal and distal endplates of the respective proximal and distal vertebrae comprising the curve. Traditionally, Cobb angles are measured manually using either a marker, straight edge, and protractor or image measurement software. The task of measuring the Cobb angle can also be represented by a function taking the spine geometry rendered using X-ray imaging as input and returning the approximate angle. Although the form of such a function may be unknown, it can be approximated using artificial neural networks (ANNs). The performance of ANNs is affected by many factors, including the choice of activation function and network architecture; however, the effects of these parameters on the accuracy of scoliotic deformity measurements are poorly understood. Therefore, the objective of this study was to systematically investigate the effect of ANN architecture and activation function on Cobb angle measurement from the coronal X-rays of scoliotic subjects. The data set for this study consisted of 609 coronal chest X-rays of scoliotic subjects divided into 481 training images and 128 test images. These data, which included labeled Cobb angle measurements, were obtained from the SpineWeb online database. In order to normalize the input data, each image was resized using bi-linear interpolation to a size of 500 × 187 pixels, and the pixel intensities were scaled to be between 0 and 1. A fully connected (dense) ANN with a fixed cost function (mean squared error), batch size (10), and learning rate (0.01) was developed using Python Version 3.7.3 and TensorFlow 1.13.1. The activation functions (sigmoid, hyperbolic tangent [tanh], or rectified linear units [ReLU]), number of hidden layers (1, 3, 5, or 10), and number of neurons per layer (10, 100, or 1000) were varied systematically to generate a total of 36 network conditions. Stochastic gradient descent with early stopping was used to train each network. Three trials were run per condition, and the final mean squared errors and mean absolute errors were averaged to quantify the network response for each condition. The network that performed the best used ReLU neurons had three hidden layers, and 100 neurons per layer. The average mean squared error of this network was 222.28 ± 30 degrees2, and the average mean absolute error was 11.96 ± 0.64 degrees. It is also notable that while most of the networks performed similarly, the networks using ReLU neurons, 10 hidden layers, and 1000 neurons per layer, and those using Tanh neurons, one hidden layer, and 10 neurons per layer performed markedly worse with average mean squared errors greater than 400 degrees2 and average mean absolute errors greater than 16 degrees. From the results of this study, it can be seen that the choice of ANN architecture and activation function has a clear impact on Cobb angle inference from coronal X-rays of scoliotic subjects.

Keywords: scoliosis, artificial neural networks, cobb angle, medical imaging

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Authors: Elena Perepelova

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As the world approaches the 4th industrial revolution, income inequality has become one of the major societal concerns. Displacement of workers by technology becomes a reality, and in return, new skills and competencies are required. More important than ever, education needs to help individuals understand the wider world around them and make global connections. The author argues for the necessity to incorporate business, economics and finance studies as a part of primary education and offer access to business education to the general population with the primary objective to understand how the world functions. The paper offers a fresh look at existing business theory through an innovative program called 'Usefulnomics'. Realizing that the subject of Economics, Finance and Business are perceived as overwhelming for a large part of the population, the author has taken a holistic approach and created a program that simplifies the definitions of the existing concepts and shifts from the traditional breakdown into subjects and specialties to a teaching method that is based exclusively on real-life example case studies and group debates, in order to better grasp the concepts and put them into context. The paper findings are the result of a two-year project and experimental work with students from UK, USA, Malaysia, Russia, and Spain. The author conducted extensive research through on-line and in-person classes and workshops as well as in-depth interviews of primary and secondary grade students to assess their understanding of what is a business, how businesses operate and the role businesses play in their communities. The findings clearly indicate that students of all ages often understood business concepts and processes only in an intuitive way, which resulted in misconceptions and gaps in knowledge. While knowledge gaps were easier to identify and correct in primary school students, as students’ age increased, the learning process became distorted by career choices, political views, and the students’ actual (or perceived) economic status. While secondary school students recognized more concepts, their real understanding was often on par with upper primary school age students. The research has also shown that lack of correct vocabulary created a strong barrier to communication and real-life application or further learning. Based on these findings, each key business concept was practiced and put into context with small groups of students in order to design the content and format which would be well accepted and understood by the target group. As a result, the final learning program package was based on case studies from daily modern life and used a wide range of examples: from popular brands and well-known companies to basic commodities. In the final stage, the content and format were put into practice in larger classrooms. The author would like to share the key findings from the research, the resulting learning program as well as present new ideas on how the program could be further enriched and adapted so schools and organizations can deliver it.

Keywords: business, finance, economics, lifelong learning, XXI century skills

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Authors: Sarah Lister, Pauline Palmer

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Research findings presented within this paper form part of a larger scale collaboration between academics at Manchester Metropolitan University and a technology company. The overarching aims of this project focus on developing a series of game-based resources to promote the teaching of aspects of mathematics through a second language (L2) in primary schools. This study explores the potential of game-based learning (GBL) as a dynamic way to engage and motivate learners, making learning fun and purposeful. The research examines the capacity of GBL resources to provide a meaningful and purposeful context for CLIL. GBL is a powerful learning environment and acts as an effective vehicle to promote the learning of mathematics through an L2. The fun element of GBL can minimise stress and anxiety associated with mathematics and L2 learning that can create barriers. GBL provides one of the few safe domains where it is acceptable for learners to fail. Games can provide a life-enhancing experience for learners, revolutionizing the routinized ways of learning through fusing learning and play. This study argues that playing games requires learners to think creatively to solve mathematical problems, using the L2 in order to progress, which can be associated with the development of higher-order thinking skills and independent learning. GBL requires learners to engage appropriate cognitive processes with increased speed of processing, sensitivity to environmental inputs, or flexibility in allocating cognitive and perceptual resources. At surface level, GBL resources provide opportunities for learners to learn to do things. Games that fuse subject content and appropriate learning objectives have the potential to make learning academic subjects more learner-centered, promote learner autonomy, easier, more enjoyable, more stimulating and engaging and therefore, more effective. Data includes observations of the children playing the games and follow up group interviews. Given that learning as a cognitive event cannot be directly observed or measured. A Cognitive Discourse Functions (CDF) construct was used to frame the research, to map the development of learners’ conceptual understanding in an L2 context and as a framework to observe the discursive interactions that occur learner to learner and between learner and teacher. Cognitively, the children were required to engage with mathematical content, concepts and language to make decisions quickly, to engage with the gameplay to reason, solve and overcome problems and learn through experimentation. The visual elements of the games supported the learning of new concepts. Children recognised the value of the games to consolidate their mathematical thinking and develop their understanding of new ideas. The games afforded them time to think and reflect. The teachers affirmed that the games provided meaningful opportunities for the learners to practise the language. The findings of this research support the view that using the game-based resources supported children’s grasp of mathematical ideas and their confidence and ability to use the L2. Engaging with the content and language through the games led to deeper learning.

Keywords: CLIL, gaming, language, mathematics

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Authors: Sebastian Hiergeist

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It is a common view that Unmanned Aerial Vehicles (UAV) tend to migrate into the civil airspace. This trend is challenging UAV manufacturer in plenty ways, as there come up a lot of new requirements and functional aspects. On the higher application levels, this might be collision detection and avoidance and similar features, whereas all these functions only act as input for the flight control components of the aircraft. The flight control computer (FCC) is the central component when it comes up to ensure a continuous safe flight and landing. As these systems are flight critical, they have to be built up redundantly to be able to provide a Fail-Operational behavior. Recent architectural approaches of FCCs used in UAV systems are often based on very simple microprocessors in combination with proprietary Application-Specific Integrated Circuit (ASIC) or Field Programmable Gate Array (FPGA) extensions implementing the whole redundancy functionality. In the future, such simple microprocessors may not be available anymore as they are more and more replaced by higher sophisticated System on Chip (SoC). As the avionic industry cannot provide enough market power to significantly influence the development of new semiconductor products, the use of solutions from foreign markets is almost inevitable. Products stemming from the industrial market developed according to IEC 61508, or automotive SoCs, according to ISO 26262, can be seen as candidates as they have been developed for similar environments. Current available SoC from the industrial or automotive sector provides quite a broad selection of interfaces like, i.e., Ethernet, SPI or FlexRay, that might come into account for the implementation of a redundancy network. In this context, possible network architectures shall be investigated which could be established by using the interfaces stated above. Of importance here is the avoidance of any single point of failures, as well as a proper segregation in distinct fault containment regions. The performed analysis is supported by the use of guidelines, published by the aviation authorities (FAA and EASA), on the reliability of data networks. The main focus clearly lies on the reachable level of safety, but also other aspects like performance and determinism play an important role and are considered in the research. Due to the further increase in design complexity of recent and future SoCs, also the risk of design errors, which might lead to common mode faults, increases. Thus in the context of this work also the aspect of dissimilarity will be considered to limit the effect of design errors. To achieve this, the work is limited to broadly available interfaces available in products from the most common silicon manufacturer. The resulting work shall support the design of future UAV FCCs by giving a guideline on building up a redundancy network between SoCs, solely using on board interfaces. Therefore the author will provide a detailed usability analysis on available interfaces provided by recent SoC solutions, suggestions on possible redundancy architectures based on these interfaces and an assessment of the most relevant characteristics of the suggested network architectures, like e.g. safety or performance.

Keywords: redundancy, System-on-Chip, UAV, flight control computer (FCC)

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Authors: Rebeca P Ramos-Bueno, Maria Jose Gonzalez-Fernandez, Rosa M. Moreno-Zamora, Antonia Barros Heras, Yolanda Serrano Alonso, Carolina Sanchez Barranco

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Insects are a new source of fatty acids (FA), so they are considered a sustainable and environmentally friendly alternative for both animal feed and the human diet, and furthermore, their harvesting/rearing require a low-tech and low capital investment. For that reason, lipids obtained by insect breeding open interesting possibilities with alimentary and industrial purposes, i.e., the production of biodiesel. Particularly, certain insect species, especially during the larval stage, contain high proportions of fat which is highly dependent on their feed and stage of development. Among them, Hermetia illucens larvae can be bred on food wastes to produce fat- and protein-rich raw materials for food by-product management. So, insects can act as excellent bioconverters of organic waste to nutrient-rich materials. In this regard, the aim of the study was to evaluate the effects of fruit byproducts on the FA compositions of Tenebrio molitor, Zophoba morio, and H. illucens larvae. Firstly, oil was extracted with the green solvent ethyl acetate, and FA methyl ester was obtained and analyzed by GC to show the FA profile. In addition, the triacylglycerol (TAG) profile was obtained by HPLC. Dehydrated watermelon, tomato, and papaya by-products, as well as wheat-based control feed, were assayed. High FA content was reached by Z. morio larvae fed with all fruits; however, no differences were shown in lipid profile with any change. It is worth highlighting that both Z. morio and H. illucens could be selected as the best candidates for biodiesel production due to their high content of saturated FA. On the other hand, T. molitor larvae showed a higher content of monounsaturated FA than control larvae, whereas the n-6 polyunsaturated FA content decreased in larvae fed with fruits. This result indicates that the improvement of the FA profile of Tenebrio can depend on both the type of feeding and the intended use. The lipid profile of H. illucens larvae fed with papaya and tomato showed a slight increase in the content of α-linoleic acid (ALA, 18:3n3). This FA is the precursor of docosahexaenoic acid (DHA, 22:6n3), which plays an important role as a component of structural lipids in cell membranes as well as in the synthesis of eicosanoids, protecting and resolving. Also, it was evaluated the TAG profile of Z. morio larvae due to their highest oil content. The results showed a high oleic acid (OA, 18:1n9) content, which displays modulatory effects in a wide range of physiological functions, having anti-inflammatory and anti-atherogenic properties. In conclusion, this study clearly shows that Z. morio and H. illucens larvae constitute an alternative source of OA- and ALA-rich oils, respectively, which can be devoted for food use, as well as for using in the food and pharmaceutical industries, with agronomic implications. Finally, although the profile of Z. morio was not improved with fruit feeding, this kind of feeding could be used due to its low environmental impact.

Keywords: fatty acids, fruit byproducts, Hermetia illucens, Zophoba morio, Tenebrio molitor, insect rearing

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Authors: R. P. Naik, A. K. Rakshit

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In recent years, with the rapid development in science and technology, people have increasing requirements on uses of clothing for new functions, which contributes to opportunities for further development and incorporation of new technologies along with novel materials. In this context, textiles are of fast decalescence or fast heat radiation media as per as comfort accountability of textile articles are concern. The microstructure and texture of textiles play a vital role in determining the heat-moisture comfort level of the human body because clothing serves as a barrier to the outside environment and a transporter of heat and moisture from the body to the surrounding environment to keep thermal balance between body heat produced and body heat loss. The main bottleneck which is associated with textile materials to be successful as thermal insulation materials can be enumerated as; firstly, high loft or bulkiness of material so as to provide predetermined amount of insulation by ensuring sufficient trapping of air. Secondly, the insulation depends on forced convection; such convective heat loss cannot be prevented by textile material. Third is that the textile alone cannot reach the level of thermal conductivity lower than 0.025 W/ m.k of air. Perhaps, nano-fibers can do so, but still, mass production and cost-effectiveness is a problem. Finally, such high loft materials for thermal insulation becomes heavier and uneasy to manage especially when required to carry over a body. The proposed works aim at developing lightweight effective thermal insulation textiles in combination with nanoporous silica-gel which provides the fundamental basis for the optimization of material properties to achieve good performance of the clothing system. This flexible nonwoven silica-gel composites fabric in intact monolith was successfully developed by reinforcing SiO2-gel in thermal bonded nonwoven fabric via sol-gel processing. Ambient Pressure Drying method is opted for silica gel preparation for cost-effective manufacturing. The formed structure of the nonwoven / SiO₂ -gel composites were analyzed, and the transfer properties were measured. The effects of structure and fibre on the thermal properties of the SiO₂-gel composites were evaluated. Samples are then tested against untreated samples of same GSM in order to study the effect of SiO₂-gel application on various properties of nonwoven fabric. The nonwoven fabric composites reinforced with aerogel showed intact monolith structure were also analyzed for their surface structure, functional group present, microscopic images. Developed product reveals a significant reduction in pores' size and air permeability than the conventional nonwoven fabric. Composite made from polyester fibre with lower GSM shows lowest thermal conductivity. Results obtained were statistically analyzed by using STATISTICA-6 software for their level of significance. Univariate tests of significance for various parameters are practiced which gives the P value for analyzing significance level along with that regression summary for dependent variable are also studied to obtain correlation coefficient.

Keywords: silica-gel, heat insulation, nonwoven fabric, thermal barrier clothing

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

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

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

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Authors: M. Saard, A. Kolk, K. Sepp, L. Pertens, L. Reinart, C. Kööp

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Introduction: Social competence is often impaired in children with acquired brain injury (ABI), but evidence-based rehabilitation for social skills has remained undeveloped. Modern technology-based methods create effective and safe learning environments for pediatric social skills remediation. The aim of the study was to implement our structured model of neuro rehab for socio-cognitive deficit using multitouch-multiuser tabletop (MMT) computer-based platforms and virtual reality (VR) technology. Methods: 40 children aged 8-13 years (yrs) have participated in the pilot study: 30 with ABI -epilepsy, traumatic brain injury and/or tic disorder- and 10 healthy age-matched controls. From the patients, 12 have completed the training (M = 11.10 yrs, SD = 1.543) and 20 are still in training or in the waiting-list group (M = 10.69 yrs, SD = 1.704). All children performed the first individual and paired assessments. For patients, second evaluations were performed after the intervention period. Two interactive applications were implemented into rehabilitation design: Snowflake software on MMT tabletop and NoProblem on DiamondTouch Table (DTT), which allowed paired training (2 children at once). Also, in individual training sessions, HTC Vive VR device was used with VR metaphors of difficult social situations to treat social anxiety and train social skills. Results: At baseline (B) evaluations, patients had higher deficits in executive functions on the BRIEF parents’ questionnaire (M = 117, SD = 23.594) compared to healthy controls (M = 22, SD = 18.385). The most impaired components of social competence were emotion recognition, Theory of Mind skills (ToM), cooperation, verbal/non-verbal communication, and pragmatics (Friendship Observation Scale scores only 25-50% out of 100% for patients). In Sentence Completion Task and Spence Anxiety Scale, the patients reported a lack of friends, behavioral problems, bullying in school, and social anxiety. Outcome evaluations: Snowflake on MMT improved executive and cooperation skills and DTT developed communication skills, metacognitive skills, and coping. VR, video modelling and role-plays improved social attention, emotional attitude, gestural behaviors, and decreased social anxiety. NEPSY-II showed improvement in Affect Recognition [B = 7, SD = 5.01 vs outcome (O) = 10, SD = 5.85], Verbal ToM (B = 8, SD = 3.06 vs O = 10, SD = 4.08), Contextual ToM (B = 8, SD = 3.15 vs O = 11, SD = 2.87). ToM Stories test showed an improved understanding of Intentional Lying (B = 7, SD = 2.20 vs O = 10, SD = 0.50), and Sarcasm (B=6, SD = 2.20 vs O = 7, SD = 2.50). Conclusion: Neurorehabilitation based on the Structured Model of Neurorehab for Socio-Cognitive Deficit in children with ABI were effective in social skills remediation. The model helps to understand theoretical connections between components of social competence and modern interactive computerized platforms. We encourage therapists to implement these next-generation devices into the rehabilitation process as MMT and VR interfaces are motivating for children, thus ensuring good compliance. Improving children’s social skills is important for their and their families’ quality of life and social capital.

Keywords: acquired brain injury, children, social skills deficit, technology-based neurorehabilitation

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Authors: Yi Wang, Greg Hearn, Terry Flew

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In order to understand how cultural innovation occurs, this paper explores the interaction in Shenzhen of China between universities, creative industries, and government in creative economic using the Triple Helix framework. During the past two decades, Triple Helix has been recognized as a new theory of innovation to inform and guide policy-making in national and regional development. Universities and governments around the world, especially in developing countries, have taken actions to strengthen connections with creative industries to develop regional economies. To date research based on the Triple Helix model has focused primarily on Science and Technology collaborations, largely ignoring other fields. Hence, there is an opportunity for work to be done in seeking to better understand how the Triple Helix framework might apply in the field of creative industries and what knowledge might be gleaned from such an undertaking. Since the late 1990s, the concept of ‘creative industries’ has been introduced as policy and academic discourse. The development of creative industries policy by city agencies has improved city wealth creation and economic capital. It claims to generate a ‘new economy’ of enterprise dynamics and activities for urban renewal through the arts and digital media, via knowledge transfer in knowledge-based economies. Creative industries also involve commercial inputs to the creative economy, to dynamically reshape the city into an innovative culture. In particular, this paper will concentrate on creative spaces (incubators, digital tech parks, maker spaces, art hubs) where academic, industry and government interact. China has sought to enhance the brand of their manufacturing industry in cultural policy. It aims to transfer the image of ‘Made in China’ to ‘Created in China’ as well as to give Chinese brands more international competitiveness in a global economy. Shenzhen is a notable example in China as an international knowledge-based city following this path. In 2009, the Shenzhen Municipal Government proposed the city slogan ‘Build a Leading Cultural City”’ to show the ambition of government’s strong will to develop Shenzhen’s cultural capacity and creativity. The vision of Shenzhen is to become a cultural innovation center, a regional cultural center and an international cultural city. However, there has been a lack of attention to the triple helix interactions in the creative industries in China. In particular, there is limited knowledge about how interactions in creative spaces co-location within triple helix networks significantly influence city based innovation. That is, the roles of participating institutions need to be better understood. Thus, this paper discusses the interplay between university, creative industries and government in Shenzhen. Secondary analysis and documentary analysis will be used as methods in an effort to practically ground and illustrate this theoretical framework. Furthermore, this paper explores how are creative spaces being used to implement Triple Helix in creative industries. In particular, the new combination of resources generated from the synthesized consolidation and interactions through the institutions. This study will thus provide an innovative lens to understand the components, relationships and functions that exist within creative spaces by applying Triple Helix framework to the creative industries.

Keywords: cultural policy, creative industries, creative city, triple Helix

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Authors: Lijuan Li

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Nitric oxide (NO) has become a fascinating entity in biological chemistry over the past few years. It is a gaseous lipophilic radical molecule that plays important roles in several physiological and pathophysiological processes in mammals, including activating the immune response, serving as a neurotransmitter, regulating the cardiovascular system, and acting as an endothelium-derived relaxing factor. NO functions in eukaryotes both as a signal molecule at nanomolar concentrations and as a cytotoxic agent at micromolar concentrations. The latter arises from the ability of NO to react readily with a variety of cellular targets leading to thiol S-nitrosation, amino acid N-nitrosation, and nitrosative DNA damage. Nitric oxide can readily bind to metals to give metal-nitrosyl (M-NO) complexes. Some of these species are known to play roles in biological NO storage and transport. These complexes have different biological, photochemical, or spectroscopic properties due to distinctive structural features. These recent discoveries have spawned a great interest in the development of transition metal complexes containing NO, particularly its iron complexes that are central to the role of nitric oxide in the body. Spectroscopic evidence would appear to implicate species of “Fe(NO)2+” type in a variety of processes ranging from polymerization, carcinogenesis, to nitric oxide stores. Our research focuses on isolation and structural studies of non-heme iron nitrosyls that mimic biologically active compounds and can potentially be used for anticancer drug therapy. We have shown that reactions between Fe(NO)2(CO)2 and a series of imidazoles generated new non-heme iron nitrosyls of the form Fe(NO)2(L)2 [L = imidazole, 1-methylimidazole, 4-methylimidazole, benzimidazole, 5,6-dimethylbenzimidazole, and L-histidine] and a tetrameric cluster of [Fe(NO)2(L)]4 (L=Im, 4-MeIm, BzIm, and Me2BzIm), resulted from the interactions of Fe(NO)2 with a series of substituted imidazoles was prepared. Recently, a series of sulfur bridged iron di nitrosyl complexes with the general formula of [Fe(µ-RS)(NO)2]2 (R = n-Pr, t-Bu, 6-methyl-2-pyridyl, and 4,6-dimethyl-2-pyrimidyl), were synthesized by the reaction of Fe(NO)2(CO)2 with thiols or thiolates. Their structures and properties were studied by IR, UV-vis, 1H-NMR, EPR, electrochemistry, X-ray diffraction analysis and DFT calculations. IR spectra of these complexes display one weak and two strong NO stretching frequencies (νNO) in solution, but only two strong νNO in solid. DFT calculations suggest that two spatial isomers of these complexes bear 3 Kcal energy difference in solution. The paramagnetic complexes [Fe2(µ-RS)2(NO)4]-, have also been investigated by EPR spectroscopy. Interestingly, the EPR spectra of complexes exhibit an isotropic signal of g = 1.998 - 2.004 without hyperfine splitting. The observations are consistent with the results of calculations, which reveal that the unpaired electron dominantly delocalize over the two sulfur and two iron atoms. The difference of the g values between the reduced form of iron-sulfur clusters and the typical monomeric di nitrosyl iron complexes is explained, for the first time, by of the difference in unpaired electron distributions between the two types of complexes, which provides the theoretical basis for the use of g value as a spectroscopic tool to differentiate these biologically active complexes.

Keywords: di nitrosyl iron complex, metal nitrosyl, non-heme iron, nitric oxide

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Authors: M. Testa, L. Peotta, S. Giusiano, B. Lazzolino, U. Manera, A. Canosa, M. Grassano, F. Palumbo, A. Bombaci, S. Cabras, F. Di Pede, L. Solero, E. Matteoni, C. Moglia, A. Calvo, A. Chio

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Amyotrophic Lateral Sclerosis (ALS) is a neurodegenerative disease of adulthood, which primarily affects the central nervous system and is characterized by progressive bilateral degeneration of motor neurons. The degeneration processes in ALS extend far beyond the neurons of the motor system, and affects cognition, behaviour and language. To outline the prevalence of language deficits in an ALS cohort and explore their profile along with demographic and neuropsychological data. A full neuropsychological battery and language assessment was administered to 56 ALS patients. Neuropsychological assessment included tests of executive functioning, verbal fluency, social cognition and memory. Language was assessed using tests for verbal comprehension, production and pragmatics. Patients were cognitively classified following the Revised Consensus Criteria and divided in three groups showing different levels of language deficits: group 1 - no language deficit; group 2 - one language deficit; group 3 - two or more language deficits. Chi-square for independence and non-parametric measures to compare groups were applied. Nearly half of ALS-CN patients (48%) reported one language test under the clinical cut-off, and only 13% of patents classified as ALS-CI showed no language deficits, while the rest 87% of ALS-CI reported two or more language deficits. ALS-BI and ALS-CBI cases all reported two or more language deficits. Deficits in production and in comprehension appeared more frequent in ALS-CI patients (p=0.011, p=0.003 respectively), with a higher percentage of comprehension deficits (83%). Nearly all ALS-CI reported at least one deficit in pragmatic abilities (96%) and all ALS-BI and ALS-CBI patients showed pragmatic deficits. Males showed higher percentage of pragmatic deficits (97%, p=0.007). No significant differences in language deficits have been found between bulbar and spinal onset. Months from onset and level of impairment at testing (ALS-FRS total score) were not significantly different between levels and type of language impairment. Age and education were significantly higher for cases showing no deficits in comprehension and pragmatics and in the group showing no language deficits. Comparing performances at neuropsychological tests among the three levels of language deficits, no significant differences in neuropsychological performances were found between group 1 and 2; compared to group 1, group 3 appeared to decay specifically on executive testing, verbal/visuospatial learning, and social cognition. Compared to group 2, group 3 showed worse performances specifically in tests of working memory and attention. Language deficits have found to be spread in our sample, encompassing verbal comprehension, production and pragmatics. Our study reveals that also cognitive intact patients (ALS-CN) showed at least one language deficit in 48% of cases. Pragmatic domain is the most compromised (84% of the total sample), present in nearly all ALS-CI (96%), likely due to the influence of executive impairment. Lower age and higher education seem to preserve comprehension, pragmatics and presence of language deficits. Finally, executive functions, verbal/visuospatial learning and social cognition differentiate the group with no language deficits from the group with a clinical language impairment (group 3), while attention and working memory differentiate the group with one language deficit from the clinical impaired group.

Keywords: amyotrophic lateral sclerosis, language assessment, neuropsychological assessment, language deficit

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Authors: James Cairns, Marco Vezza, Richard Green, Donald MacVicar

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Significant legislative changes are set to revolutionise the commercial shipping industry. Upcoming emissions restrictions will force operators to look at technologies that can improve the efficiency of their vessels -reducing fuel consumption and emissions. A device which may help in this challenge is the Ship Wind-Assisted Propulsion system (SWAP), an actively controlled aerofoil mounted vertically on the deck of a ship. The device functions in a similar manner to a sail on a yacht, whereby the aerodynamic forces generated by the sail reach an equilibrium with the hydrodynamic forces on the hull and a forward velocity results. Numerical and experimental testing of the SWAP device is presented in this study. Circulation control takes the form of a co-flow jet aerofoil, utilising both blowing from the leading edge and suction from the trailing edge. A jet at the leading edge uses the Coanda effect to energise the boundary layer in order to delay flow separation and create high lift with low drag. The SWAP concept has been originated by the research and development team at SMAR Azure Ltd. The device will be retrofitted to existing ships so that a component of the aerodynamic forces acts forward and partially reduces the reliance on existing propulsion systems. Wind tunnel tests have been carried out at the de Havilland wind tunnel at the University of Glasgow on a 1:20 scale model of this system. The tests aim to understand the airflow characteristics around the aerofoil and investigate the approximate lift and drag coefficients that an early iteration of the SWAP device may produce. The data exhibits clear trends of increasing lift as injection momentum increases, with critical flow attachment points being identified at specific combinations of jet momentum coefficient, Cµ, and angle of attack, AOA. Various combinations of flow conditions were tested, with the jet momentum coefficient ranging from 0 to 0.7 and the AOA ranging from 0° to 35°. The Reynolds number across the tested conditions ranged from 80,000 to 240,000. Comparisons between 2D computational fluid dynamics (CFD) simulations and the experimental data are presented for multiple Reynolds-Averaged Navier-Stokes (RANS) turbulence models in the form of normalised surface pressure comparisons. These show good agreement for most of the tested cases. However, certain simulation conditions exhibited a well-documented shortcoming of RANS-based turbulence models for circulation control flows and over-predicted surface pressures and lift coefficient for fully attached flow cases. Work must be continued in finding an all-encompassing modelling approach which predicts surface pressures well for all combinations of jet injection momentum and AOA.

Keywords: CFD, circulation control, Coanda, turbo wing sail, wind tunnel

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Authors: B. Atika, S. Lehmann, E. Wimmer

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The defense strategy is very common in the insect world. Defensive substances play a wide variety of functions for beetles, such as repellents, toxicants, insecticides, and antimicrobics. Beetles react to predators, invaders, and parasitic microbes with the release of toxic and repellent substances. Defensive substances are directed against a large array of potential target organisms or may function for boiling bombardment or as surfactants. Usually, Coleoptera biosynthesize and store their defensive compounds in a complex secretory organ, known as odoriferous defensive stink glands. The red flour beetle, Tribolium castaneum (Coleoptera: Tenebrionidae), uses these glands to produce antimicrobial p-benzoquinones and 1-alkenes. In the past, the morphology of stink gland has been studied in detail in tenebrionid beetles; however, very little is known about the genes that are involved in the production of gland secretion. In this study, we studied a subset of genes that are essential for the benzoquinone production in red flour beetle. In the first phase, we selected 74 potential candidate genes from a genome-wide RNA interference (RNAi) knockdown screen named 'iBeetle.' All these 74 candidate genes were functionally characterized by RNAi-mediated gene knockdown. Therefore, they were selected for a subsequent gas chromatography-mass spectrometry (GC-MS) analysis of secretion volatiles in respective RNAi knockdown glands. 33 of them were observed to alter the phenotype of stink gland. In the GC-MS analysis, 7 candidate genes were noted to display a strongly altered gland, in terms of secretion color and chemical composition, upon knockdown, showing their key role in the biosynthesis of gland secretion. Morphologically altered stink glands were found for odorant receptor and protein kinase superfamily. Subsequent GC-MS analysis of secretion volatiles revealed reduced benzoquinone levels in LIM domain, PDZ domain, PBP/GOBP family knockdowns and a complete lack of benzoquinones in the knockdown of sulfatase-modifying factor enzyme 1, sulfate transporter family. Based on stink gland transcriptome data, we analyzed the function of sulfatase-modifying factor enzyme 1 and sulfate transporter family via RNAi-mediated gene knockdowns, GC-MS, in situ hybridization, and enzymatic activity assays. Morphologically altered stink glands were noted in knockdown of both these genes. Furthermore, GC-MS analysis of secretion volatiles showed a complete lack of benzoquinones in the knockdown of these two genes. In situ hybridization showed that these two genes are expressed around the vesicle of certain subgroup of secretory stink gland cells. Enzymatic activity assays on stink gland tissue showed that these genes are involved in p-benzoquinone biosynthesis. These results suggest that sulfatase-modifying factor enzyme 1 and sulfate transporter family play a role specifically in benzoquinone biosynthesis in red flour beetles.

Keywords: Red Flour Beetle, defensive stink gland, benzoquinones, sulfate transporter, sulfatase-modifying factor enzyme 1

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Authors: Jose CanIzares

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In the last decades, resilience has received much attention in relation to understanding and managing new forms of risk, especially in the context of urban adaptation to climate change. There are abundant concerns, however, on how to best interpret resilience and related ideas, and on whether they can guide ethically appropriate risk-related or adaptation efforts. Narrative creation and framing are critical steps in shaping public discussion and policy in large-scale interventions, since they favor or inhibit early decision and interpretation habits, which can be morally sensitive and then become persistent on time. This article adds to such framing process by contesting a conventional narrative on resilience and offering an alternative one. Conventionally, present ideas on resilience are traced to the work of ecologist C. S. Holling, especially to his article Resilience and Stability in Ecosystems. This article is usually portrayed as a contribution of complex systems thinking to theoretical ecology, where Holling appeals to resilience in order to challenge received views on ecosystem stability and the diversity-stability hypothesis. In this regard, resilience is construed as a “purely scientific”, precise and descriptive concept, denoting a complex property that allows ecosystems to persist, or to maintain functions, after disturbance. Yet, these formal features of resilience supposedly changed with Holling’s later work in the 90s, where, it is argued, Holling begun to use resilience as a more pragmatic “boundary term”, aimed at unifying transdisciplinary research about risks, ecological or otherwise, and at articulating public debate and governance strategies on the issue. In the conventional story, increased vagueness and degrees of normativity are the price to pay for this conceptual shift, which has made the term more widely usable, but also incompatible with scientific purposes and morally problematic (if not completely objectionable). This paper builds on a detailed analysis of Holling’s early work to propose an alternative narrative. The study will show that the “complexity turn” has often entangled theoretical and pragmatic aims. Accordingly, Holling’s primary aim was to fight what he termed “pathologies of natural resource management” or “pathologies of command and control management”, and so, the terms of his reform of ecosystem science are partly subordinate to the details of his proposal for reforming the management sciences. As regards resilience, Holling used it as a polysemous, ambiguous and normative term: sometimes, as an instrumental value that is closely related to various stability concepts; other times, and more crucially, as an intrinsic value and a tool for attacking efficiency and instrumentalism in management. This narrative reveals the limitations of its conventional alternative and has several practical advantages. It captures well the structure and purposes of Holling’s project, and the various roles of resilience in it. It helps to link Holling’s early work with other philosophical and ideological shifts at work in the 70s. It highlights the currency of Holling’s early work for present research and action in fields such as risk and climate adaptation. And it draws attention to morally relevant aspects of resilience that the conventional narrative neglects.

Keywords: resilience, complexity turn, risk management, positivistic, framing

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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: Bernice Jeanne Lottering, Yi-Wen Lin

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Introduction: Chronic pain has a definitive lack of objective parameters in the measurement and treatment efficacy of diseases such as Fibromyalgia (FM). Persistent widespread pain and generalized tenderness are the characteristic symptoms affecting a large majority of the global population, particularly females. This disease has indicated a refractory tendency to conventional treatment ventures, largely resultant from a lack of etiological and pathogenic understanding of the disease development. Emerging evidence indicates that the central nervous system (CNS) plays a critical role in the amplification of pain signals and the neurotransmitters associated therewith. Various stimuli have been found to activate the channels existent on nociceptor terminals, thereby actuating nociceptive impulses along the pain pathways. The transient receptor potential vanalloid 1 (TRPV1) channel functions as a molecular integrator for numerous sensory inputs, such as nociception, and was explored in the current study. Current intervention approaches face a multitude challenges, ranging from effective therapeutic interventions to the limitation of pathognomonic criteria resultant from incomplete understanding and partial evidence on the mechanisms of action of FM. It remains unclear whether electroacupuncture (EA) plays an integral role in the functioning of the TRPV1 pathway, and whether or not it can reduce the chronic pain induced by FM. Aims: The aim of this study was to explore the mechanisms underlying the activation and modulation of the TRPV1 channel pathway in a cold stress model of FM applied to a murine model. Furthermore, the effect of EA in the treatment of mechanical and thermal pain, as expressed in FM was also to be investigated. Methods: 18 C57BL/6 wild type and 6 TRPV1 knockout (KO) mice, aged 8-12 weeks, were exposed to an intermittent cold stress-induced fibromyalgia-like pain model, with or without EA treatment at ZusanLi ST36 (2Hz/20min) on day 3 to 5. Von Frey and Hargreaves behaviour tests were implemented in order to analyze the mechanical and thermal pain thresholds on day 0, 3 and 5 in control group (C), FM group (FM), FM mice with EA treated group (FM + EA) and FM in KO group. Results: An increase in mechanical and thermal hyperalgesia was observed in the FM, EA and KO groups when compared to the control group. This initial increase was reduced in the EA group, which directs focus at the treatment efficacy of EA in nociceptive sensitization, and the analgesic effect EA has attenuating FM associated pain. Discussion: An increase in the nociceptive sensitization was observed through higher withdrawal thresholds in the von Frey mechanical test and the Hargreaves thermal test. TRPV1 function in mice has been scientifically associated with these nociceptive conduits, and the increased behaviour test results suggest that TRPV1 upregulation is central to the FM induced hyperalgesia. This data was supported by the decrease in sensitivity observed in results of the TRPV1 KO group. Moreover, the treatment of EA showed a decrease in this FM induced nociceptive sensitization, suggesting TRPV1 upregulation and overexpression can be attenuated by EA at bilateral ST36. This evidence compellingly implies that the analgesic effect of EA is associated with TRPV1 downregulation.

Keywords: fibromyalgia, electroacupuncture, TRPV1, nociception

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Authors: Anna Höfer, Johannes Herrmann, Patrick Meybohm, Christopher Lotz

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Mitochondria are pivotal for energy supply and regulation of cellular functions. Deficiencies of mitochondrial metabolism have been implicated in diverse stressful conditions including infections. Platelets are key mediators for thrombo-inflammation during development and resolution of acute respiratory distress syndrome (ARDS). Previous data point to an exhausted platelet phenotype in critically-ill patients with coronavirus 19 disease (COVID-19) impacting the course of disease. The objective of this work was to characterize platelet mitochondrial metabolism in patients suffering from COVID-19 ARDSA longitudinal analysis of platelet mitochondrial metabolism in 24 patients with COVID-19 induced ARDS compared to 35 healthy controls (ctrl) was performed. Blood samples were analyzed at two time points (t1=day 1; t2=day 5-7 after study inclusion). The activity of mitochondrial citrate synthase was photometrically measured. The impact of oxidative stress on mitochondrial permeability was assessed by a photometric calcium-induced swelling assay and the activity of superoxide dismutase (SOD) by a SOD assay kit. The amount of protein carbonylation and the activity of mitochondria complexes I-IV were photometrically determined. Levels of interleukins (IL)-1α, IL-1β and tumor necrosis factor (TNF-) α were measured by a Multiplex assay kit. Median age was 54 years, 63 % were male and BMI was 29.8 kg/m2. SOFA (12; IQR: 10-15) and APACHE II (27; IQR: 24-30) indicated critical illness. Median Murray Score was 3.4 (IQR: 2.8-3.4), 21/24 (88%) required mechanical ventilation and V-V ECMO support in 14/24 (58%). Platelet counts in ARDS did not change during ICU stay (t1: 212 vs. t2: 209 x109/L). However, mean platelet volume (MPV) significantly increased (t1: 10.6 vs. t2: 11.9 fL; p<0.0001). Citrate synthase activity showed no significant differences between ctrl and ARDS patients. Calcium induced swelling was more pronounced in patients at t1 compared to t2 and to ctrl (50µM; t1: 0.006 vs. ctrl: 0.016 ΔOD; p=0.001). The amount of protein carbonylation as marker for irreversible proteomic modification constantly increased during ICU stay and compared to ctrl., without reaching significance. In parallel, superoxid dismutase activity gradually declined during ICU treatment vs. ctrl (t2: - 29 vs. ctrl.: - 17 %; p=0.0464). Complex I analysis revealed significantly stronger activity in ARDS vs. ctrl. (t1: 0.633 vs. ctrl.: 0.415 ΔOD; p=0.0086). There were no significant differences in complex II, III or IV activity in platelets from ARDS patients compared to ctrl. IL-18 constantly increased during the observation period without reaching significance. IL-1α and TNF-α did not differ from ctrl. However, IL-1β levels were significantly elevated in ARDS (t1: 16.8; t2: 16.6 vs. ctrl.: 12.4 pg/mL; p1=0.0335, p2=0.0032). This study reveals new insights in platelet mitochondrial metabolism during COVID-19 caused ARDS. it data point towards enhanced platelet activity with a pronounced turnover rate. We found increased activity of mitochondria complex I and evidence for enhanced oxidative stress. In parallel, protective mechanisms against oxidative stress were narrowed with elevated levels of IL-1β likely causing a pro-apoptotic environment. These mechanisms may contribute to platelet exhaustion in ARDS.

Keywords: acute respiratory distress syndrome (ARDS), coronavirus 19 disease (COVID-19), oxidative stress, platelet mitochondrial metabolism

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Authors: Zenaida Chitu, Roxana Bojariu, Liliana Velea, Roxana Burcea

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A number of studies show that extreme air temperature affects mortality related to cardiovascular diseases, particularly among elderly people. In Romania, the summer thermal discomfort expressed by Universal Thermal Climate Index (UTCI) is highest in the Southern part of the country, where Bucharest, the largest Romanian urban agglomeration, is also located. The urban characteristics such as high building density and reduced green areas enhance the increase of the air temperature during summer. In Bucharest, as in many other large cities, the effect of heat urban island is present and determines an increase of air temperature compared to surrounding areas. This increase is particularly important during heat wave periods in summer. In this context, the researchers performed a temperature-mortality analysis based on daily deaths related to cardiovascular diseases, recorded between 2010 and 2019 in Bucharest. The temperature-mortality relationship was modeled by applying distributed lag non-linear model (DLNM) that includes a bi-dimensional cross-basis function and flexible natural cubic spline functions with three internal knots in the 10th, 75th and 90th percentiles of the temperature distribution, for modelling both exposure-response and lagged-response dimensions. Firstly, this study applied this analysis for the present climate. Extrapolation of the exposure-response associations beyond the observed data allowed us to estimate future effects on mortality due to temperature changes under climate change scenarios and specific assumptions. We used future projections of air temperature from five numerical experiments with regional climate models included in the EURO-CORDEX initiative under the relatively moderate (RCP 4.5) and pessimistic (RCP 8.5) concentration scenarios. The results of this analysis show for RCP 8.5 an ensemble-averaged increase with 6.1% of heat-attributable mortality fraction in future in comparison with present climate (2090-2100 vs. 2010-219), corresponding to an increase of 640 deaths/year, while mortality fraction due to the cold conditions will be reduced by 2.76%, corresponding to a decrease by 288 deaths/year. When mortality data is stratified according to the age, the ensemble-averaged increase of heat-attributable mortality fraction for elderly people (> 75 years) in the future is even higher (6.5 %). These findings reveal the necessity to carefully plan urban development in Bucharest to face the public health challenges raised by the climate change. Paper Details: This work is financed by the project URCLIM which is part of ERA4CS, an ERA-NET initiated by JPI Climate, and funded by Ministry of Environment, Romania with co-funding by the European Union (Grant 690462). A part of this work performed by one of the authors has received funding from the European Union’s Horizon 2020 research and innovation programme from the project EXHAUSTION under grant agreement No 820655.

Keywords: cardiovascular diseases, climate change, extreme air temperature, mortality

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Authors: Farideh Tabatabaee Yazdi, Fereshteh Falah, Alireza Vasiee

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Introduction: Gamma-aminobutyric acid (GABA) is a non-protein amino acid that is widely present in organisms. GABA is a kind of pharmacological and biological component and its application is wide and useful. Several important physiological functions of GABA have been characterized, such as neurotransmission and induction of hypotension. GABA is also a strong secretagogue of insulin from the pancreas and effectively inhibits small airway-derived lung adenocarcinoma and tranquilizer. Many microorganisms can produce GABA, and lactic acid bacteria have been a focus of research in recent years because lactic acid bacteria possess special physiological activities and are generally regarded as safe. Among them, the Lb. Brevis produced the highest amount of GABA. The major factors affecting GABA production have been characterized, including carbon sources and glutamate concentration. The use of food industry waste to produce valuable products such as amino acids seems to be a good way to reduce production costs and prevent the waste of food resources. In a dairy factory, a high volume of sludge is produced from a separator that contains useful compounds such as growth factors, carbon, nitrogen, and organic matter that can be used by different microorganisms such as Lb.brevis as carbon and nitrogen sources. Therefore, it is a good source of GABA production. GABA is primarily formed by the irreversible α-decarboxylation reaction of L-glutamic acid or its salts, catalysed by the GAD enzyme. In the present study, this aim was achieved for the fast-growing of Lb.brevis and producing GABA, using the dairy industry sludge as a suitable growth medium. Lactobacillus Brevis strains obtained from Microbial Type Culture Collection (MTCC) were used as model strains. In order to prepare dairy sludge as a medium, sterilization should be done at 121 ° C for 15 minutes. Lb. Brevis was inoculated to the sludge media at pH=6 and incubated for 120 hours at 30 ° C. After fermentation, the supernatant solution is centrifuged and then, the GABA produced was analyzed by the Thin Layer chromatography (TLC) method qualitatively and by the high-performance liquid chromatography (HPLC) method quantitatively. By increasing the percentage of dairy sludge in the culture medium, the amount of GABA increased. Also, evaluated the growth of bacteria in this medium showed the positive effect of dairy sludge on the growth of Lb.brevis, which resulted in the production of more GABA. GABA-producing LAB offers the opportunity of developing naturally fermented health-oriented products. Although some GABA-producing LAB has been isolated to find strains suitable for different fermentations, further screening of various GABA-producing strains from LAB, especially high-yielding strains, is necessary. The production of lactic acid, bacterial gamma-aminobutyric acid, is safe and eco-friendly. The use of dairy industry waste causes enhanced environmental safety. Also provides the possibility of producing valuable compounds such as GABA. In general, dairy sludge is a suitable medium for the growth of Lactic Acid Bacteria and produce this amino acid that can reduce the final cost of it by providing carbon and nitrogen source.

Keywords: GABA, Lactobacillus, HPLC, dairy sludge

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Authors: Kenneth A. Rostowsky, Alexander S. Maher, Nahian F. Chowdhury, Andrei Irimia

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The clinical significance of cerebral microbleeds (CMBs) due to mild traumatic brain injury (mTBI) remains unclear. Here we use magnetic resonance imaging (MRI), diffusion tensor imaging (DTI) and connectomic analysis to investigate the statistical association between mTBI-related CMBs, post-TBI changes to the human connectome and neurological/cognitive deficits. This study was undertaken in agreement with US federal law (45 CFR 46) and was approved by the Institutional Review Board (IRB) of the University of Southern California (USC). Two groups, one consisting of 26 (13 females) mTBI victims and another comprising 26 (13 females) healthy control (HC) volunteers were recruited through IRB-approved procedures. The acute Glasgow Coma Scale (GCS) score was available for each mTBI victim (mean µ = 13.2; standard deviation σ = 0.4). Each HC volunteer was assigned a GCS of 15 to indicate the absence of head trauma at the time of enrollment in our study. Volunteers in the HC and mTBI groups were matched according to their sex and age (HC: µ = 67.2 years, σ = 5.62 years; mTBI: µ = 66.8 years, σ = 5.93 years). MRI [including T1- and T2-weighted volumes, gradient recalled echo (GRE)/susceptibility weighted imaging (SWI)] and gradient echo (GE) DWI volumes were acquired using the same MRI scanner type (Trio TIM, Siemens Corp.). Skull-stripping and eddy current correction were implemented. DWI volumes were processed in TrackVis (http://trackvis.org) and 3D Slicer (http://www.slicer.org). Tensors were fit to DWI data to perform DTI, and tractography streamlines were then reconstructed using deterministic tractography. A voxel classifier was used to identify image features as CMB candidates using Microbleed Anatomic Rating Scale (MARS) guidelines. For each peri-lesional DTI streamline bundle, the null hypothesis was formulated as the statement that there was no neurological or cognitive deficit associated with between-scan differences in the mean FA of DTI streamlines within each bundle. The statistical significance of each hypothesis test was calculated at the α = 0.05 level, subject to the family-wise error rate (FWER) correction for multiple comparisons. Results: In HC volunteers, the along-track analysis failed to identify statistically significant differences in the mean FA of DTI streamline bundles. In the mTBI group, significant differences in the mean FA of peri-lesional streamline bundles were found in 21 out of 26 volunteers. In those volunteers where significant differences had been found, these differences were associated with an average of ~47% of all identified CMBs (σ = 21%). In 12 out of the 21 volunteers exhibiting significant FA changes, cognitive functions (memory acquisition and retrieval, top-down control of attention, planning, judgment, cognitive aspects of decision-making) were found to have deteriorated over the six months following injury (r = -0.32, p < 0.001). Our preliminary results suggest that acute post-TBI CMBs may be associated with cognitive decline in some mTBI patients. Future research should attempt to identify mTBI patients at high risk for cognitive sequelae.

Keywords: traumatic brain injury, magnetic resonance imaging, diffusion tensor imaging, connectomics

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Authors: Brenda Ravenscroft, Ulemu Luhanga, Bev King

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A comprehensive evaluation of online learning needs to include a blend of educational design, technology use, and online instructional practices that integrate technology appropriately for developing and delivering quality online courses. Research shows that classroom-based evaluation tools do not adequately capture the dynamic relationships between content, pedagogy, and technology in online courses. Furthermore, studies suggest that using classroom evaluations for online courses yields lower than normal scores for instructors, and may affect faculty negatively in terms of administrative decisions. In 2014, the Faculty of Arts and Science at Queen’s University responded to this evidence by seeking an alternative to the university-mandated evaluation tool, which is designed for classroom learning. The Faculty is deeply engaged in e-learning, offering large variety of online courses and programs in the sciences, social sciences, humanities and arts. This paper describes the process by which a new student survey instrument for online courses was developed and piloted, the methods used to analyze the data, and the ways in which the instrument was subsequently adapted based on the results. It concludes with a critical reflection on the challenges of evaluating e-learning. The Student Evaluation of Online Teaching Effectiveness (SEOTE), developed by Arthur W. Bangert in 2004 to assess constructivist-compatible online teaching practices, provided the starting point. Modifications were made in order to allow the instrument to serve the two functions required by the university: student survey results provide the instructor with feedback to enhance their teaching, and also provide the institution with evidence of teaching quality in personnel processes. Changes were therefore made to the SEOTE to distinguish more clearly between evaluation of the instructor’s teaching and evaluation of the course design, since, in the online environment, the instructor is not necessarily the course designer. After the first pilot phase, involving 35 courses, the results were analyzed using Stobart's validity framework as a guide. This process included statistical analyses of the data to test for reliability and validity, student and instructor focus groups to ascertain the tool’s usefulness in terms of the feedback it provided, and an assessment of the utility of the results by the Faculty’s e-learning unit responsible for supporting online course design. A set of recommendations led to further modifications to the survey instrument prior to a second pilot phase involving 19 courses. Following the second pilot, statistical analyses were repeated, and more focus groups were used, this time involving deans and other decision makers to determine the usefulness of the survey results in personnel processes. As a result of this inclusive process and robust analysis, the modified SEOTE instrument is currently being considered for adoption as the standard evaluation tool for all online courses at the university. Audience members at this presentation will be stimulated to consider factors that differentiate effective evaluation of online courses from classroom-based teaching. They will gain insight into strategies for introducing a new evaluation tool in a unionized institutional environment, and methodologies for evaluating the tool itself.

Keywords: evaluation, online courses, student survey, teaching effectiveness

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Authors: Su Mon Saw, Joe Rothnagel

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Short open reading frames (sORFs) located in 5’UTR of mRNAs are known as uORFs. Characterization of uORF-encoded peptides (uPEPs) i.e., a subset of short open reading frame encoded peptides (sPEPs) and their translation regulation lead to understanding of causes of genetic disease, proteome complexity and development of treatments. Existence of uORFs within cellular proteome could be detected by LC-MS/MS. The ability of uORF to be translated into uPEP and achievement of uPEP identification will allow uPEP’s characterization, structures, functions, subcellular localization, evolutionary maintenance (conservation in human and other species) and abundance in cells. It is hypothesized that a subset of sORFs are translatable and that their encoded sPEPs are functional and are endogenously expressed contributing to the eukaryotic cellular proteome complexity. This project aimed to investigate whether sORFs encode functional peptides. Liquid chromatography-mass spectrometry (LC-MS) and bioinformatics were thus employed. Due to probable low abundance of sPEPs and small in sizes, the need for efficient peptide enrichment strategies for enriching small proteins and depleting the sub-proteome of large and abundant proteins is crucial for identifying sPEPs. Low molecular weight proteins were extracted using SDS-PAGE from Human Embryonic Kidney (HEK293) cells and Strong Cation Exchange Chromatography (SCX) from secreted HEK293 cells. Extracted proteins were digested by trypsin to peptides, which were detected by LC-MS/MS. The MS/MS data obtained was searched against Swiss-Prot using MASCOT version 2.4 to filter out known proteins, and all unmatched spectra were re-searched against human RefSeq database. ProteinPilot v5.0.1 was used to identify sPEPs by searching against human RefSeq, Vanderperre and Human Alternative Open Reading Frame (HaltORF) databases. Potential sPEPs were analyzed by bioinformatics. Since SDS PAGE electrophoresis could not separate proteins <20kDa, this could not identify sPEPs. All MASCOT-identified peptide fragments were parts of main open reading frame (mORF) by ORF Finder search and blastp search. No sPEP was detected and existence of sPEPs could not be identified in this study. 13 translated sORFs in HEK293 cells by mass spectrometry in previous studies were characterized by bioinformatics. Identified sPEPs from previous studies were <100 amino acids and <15 kDa. Bioinformatics results showed that sORFs are translated to sPEPs and contribute to proteome complexity. uPEP translated from uORF of SLC35A4 was strongly conserved in human and mouse while uPEP translated from uORF of MKKS was strongly conserved in human and Rhesus monkey. Cross-species conserved uORFs in association with protein translation strongly suggest evolutionary maintenance of coding sequence and indicate probable functional expression of peptides encoded within these uORFs. Translation of sORFs was confirmed by mass spectrometry and sPEPs were characterized with bioinformatics.

Keywords: bioinformatics, HEK293 cells, liquid chromatography-mass spectrometry, ProteinPilot, Strong Cation Exchange Chromatography, SDS-PAGE, sPEPs

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Authors: Mark Hocking

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There is concern about the potential impact environmental flooding may have on groundwater levels and salinity processes in the Murray-Darling Basin. A study was undertaken to determine if environmental flooding of the Gunbower Forest has an impact on groundwater level and salinity which is in Victoria, Australia. To assess the impact, Impulse Response Functions (IRFs) are applied to time series groundwater monitoring well data in the area surrounding Gunbower Forest. It is found that rainfall is the primary driver of seasonal water table fluctuation, and the Murray River water level is a secondary contributor to the water table fluctuations. The dominant process that influenced the long-term water table level and salinity conditions is associated with pressure changes in the deep regional aquifer. The study demonstrates that groundwater level fluctuations in the vicinity of Gunbower Forest do not correlate with flooding (natural or managed). Groundwater recharge is calculated by applying the bore hydrograph method to the rainfall-attributed forcing function fluctuations. Data collected from thirty-three bores between 1990 to 2020 is processed to determine a 30-year average groundwater recharge rate. A 5% specific yield of the unconfined aquifer is assumed based on previously published data. It is found that the rainfall-attributed mean annual groundwater recharge varied between 2 mm/year and 189 mm/year with a median of 33.6 mm/year. Surface water recharge is also calculated by analysing the surface water attributed forcing function fluctuations and found to be as high as 37 mm/year, with most of the high values in the vicinity of rivers or agricultural land. There is a long-term regional aquifer declining trend where most water table bores have an average falling trend of 20 cm/year independent of rainfall over the past 30 years. It is found that the groundwater level beneath the Gunbower Forest is dominated by groundwater evapotranspiration. Evapotranspiration lowers the water table by as much as 0.5 m within the forest, thereby causing a relative groundwater level depression under the Gunbower Forest. Historical data shows that groundwater salinity in the area varies and has an electrical conductivity of up to 45 000 µS/cm (comparable to seawater). High groundwater salinity occurs both within and outside the Gunbower Forest as well as adjacent to the Murray River. Available groundwater salinity data suggests trends are generally stable; however, data quality and collection frequency could be improved. This study shows that at the majority of locations analyzed, the groundwater recharge occurred due to both rainfall and water loss from the Murray River. It is found that Deep groundwater pressures determined the base groundwater level, and the fluctuation of the deeper aquifer pressures determined the environmental interaction at the water surface. Local groundwater processes, such as high evapotranspiration rates in Gunbower Forest, have the capacity to lower the water table locally. The rise or fall of the regional aquifer water level has the greatest influence on the groundwater salinity in and around Gunbower Forest.

Keywords: groundwater data interpretation, groundwater monitoring, hydrogeology, impulse response function

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