Search results for: power system small-signal angular stability

Authors: Marta Gràcia, Ana Luisa Adam-Alcocer, Jesús M. Alvarado, Verónica Quezada, Tere Zarza, Priscila Garza

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In secondary education, integrating linguistic content and reflection on it is a crucial challenge that should be included in course plans to enhance students' oral communication competence. In secondary education classrooms, a continuum can be identified in relation to teaching methodologies: 1) the traditional teacher-dominated transmission approach, which is described as that in which teachers transmit content to students unidirectionally; 2) dialogical, bidirectional teaching approach that encourages students to adopt a critical vision of the information provided by the teacher or that is generated through students’ discussion. In this context, the EVALOE-DSS (Assessment Scale of Oral Language Teaching in the School Context-Decision Support System) digital instrument has emerged to help teachers in transforming their classes into spaces for communication, dialogue, reflection, evaluation of the learning process, teaching linguistic contents, and to develop curricular competencies. The tool includes various resources, such as a tutorial with the objectives and an initial screen for teachers to describe the class to be evaluated. One of the main resources of the digital instrument consists of 30 items-actions with three qualitative response options (green, orange, and red face emoji) grouped in five dimensions. In the context of the participation of secondary education teachers in a professional development program using EVALOE-DSS, a digital tool resource aimed to generate more participatory, interactive, dialogic classes, the objectives of the study were: 1) understanding the changes in classrooms’ dynamics and in the teachers’ strategies during their participation in the professional developmental program; 2) analyzing the impact of these changes in students’ oral language development according to their teachers; 3) Deeping on the impact of these changes in the students’ assessment of the classes and the self-assessment of oral competence; 4) knowing teachers’ assessment and reflections about their participation in the professional developmental program. Participants were ten teachers of different subjects and 250 students of secondary education (16-18 years) schools in Spain. The principal instrument used was the digital tool EVALOE-DSS. For 6 months, teachers used the digital tool to reflect on their classes, assess them (their actions and their students’ actions), make decisions, and introduce changes in their classes to be more participatory, interactive, and reflective about linguistic contents. Other collecting data instruments and techniques used during the study were: 1) a questionnaire to assess students’ oral language competence before and at the end of the study, 2) a questionnaire for students’ assessment of the characteristics of classes, 3) teachers’ meetings during the professional developmental program to reflect collaboratively on their experience, 4) questionnaire to assess teacher’s experience during their participation in the professional developmental program, 5) focus group meetings between the teachers and two researchers at the end of the study. The results showed relevant changes in teaching strategies, in the dynamics of the classes, which were more interactive, participative, dialogic and self-managed by the students. Both teachers and students agree about the progressive classes’ transformation into spaces for communication, discussion, and reflection on the language, its development, and its use as an essential instrument to develop curricular competencies.

Keywords: digital tool, individual and collaborative reflection, oral language competence, professional development program, secondary education

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Authors: Ankita Jadon, Sidi Souvi, Nathalie Girault, Denis Petitprez

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Safety requirements for Generation IV nuclear reactor designs, especially the new generation sodium-cooled fast reactors (SFR) require a risk-informed approach to model severe accidents (SA) and their consequences in case of outside release. In SFRs, aerosols are produced during a core disruptive accident when primary system sodium is ejected into the containment and burn in contact with the air; producing sodium aerosols. One of the key aspects of safety evaluation is the in-containment sodium aerosol behavior and their interaction with fission products. The study of the effects of sodium fires is essential for safety evaluation as the fire can both thermally damage the containment vessel and cause an overpressurization risk. Besides, during the fire, airborne fission product first dissolved in the primary sodium can be aerosolized or, as it can be the case for fission products, released under the gaseous form. The objective of this work is to study the interactions between sodium aerosols and fission products (Iodine, toxic and volatile, being the primary concern). Sodium fires resulting from an SA would produce aerosols consisting of sodium peroxides, hydroxides, carbonates, and bicarbonates. In addition to being toxic (in oxide form), this aerosol will then become radioactive. If such aerosols are leaked into the environment, they can pose a danger to the ecosystem. Depending on the chemical affinity of these chemical forms with fission products, the radiological consequences of an SA leading to containment leak tightness loss will also be affected. This work is split into two phases. Firstly, a method to theoretically understand the kinetics and thermodynamics of the heterogeneous reaction between sodium aerosols and fission products: I2 and HI are proposed. Ab-initio, density functional theory (DFT) calculations using Vienna ab-initio simulation package are carried out to develop an understanding of the surfaces of sodium carbonate (Na2CO3) aerosols and hence provide insight on its affinity towards iodine species. A comprehensive study of I2 and HI adsorption, as well as bicarbonate formation on the calculated lowest energy surface of Na2CO3, was performed which provided adsorption energies and description of the optimized configuration of adsorbate on the stable surface. Secondly, the heterogeneous reaction between (I2)g and Na2CO3 aerosols were investigated experimentally. To study this, (I2)g was generated by heating a permeation tube containing solid I2, and, passing it through a reaction chamber containing Na2CO3 aerosol deposit. The concentration of iodine was then measured at the exit of the reaction chamber. Preliminary observations indicate that there is an effective uptake of (I2)g on Na2CO3 surface, as suggested by our theoretical chemistry calculations. This work is the first step in addressing the gaps in knowledge of in-containment and atmospheric source term which are essential aspects of safety evaluation of SFR SA. In particular, this study is aimed to determine and characterize the radiological and chemical source term. These results will then provide useful insights for the developments of new models to be implemented in integrated computer simulation tool to analyze and evaluate SFR safety designs.

Keywords: iodine adsorption, sodium aerosols, sodium cooled reactor, DFT calculations, sodium carbonate

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Authors: Shah Rucksana Akhter Urme

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Agriculture is the main source of food for human consumption and feeding the world huge population, the pressure of food supply is increasing day by day. Undoubtedly, quality strain, improved plantation, farming technology, synthetic fertilizer, readily available irrigation, insecticides and harvesting technology are the main factors those to meet up the huge demand of food consumption all over the world. However, depended on this limited resources and excess amount of consuming lands, water, fertilizers leads to the end of the resources and severe climate effects has been left for our future generation. Agriculture is the most responsible to global warming, emitting more greenhouse gases than all other vehicles largely from nitrous oxide released by from fertilized fields, and carbon dioxide from the cutting of rain forests to grow crops . Farming is the thirstiest user of our precious water supplies and a major polluter, as runoff from fertilizers disrupts fragile lakes, rivers, and coastal ecosystems across the globe which accelerates the loss of biodiversity, crucial habitat and a major driver of wildlife extinction. It is needless to say that we have to more concern on how we can save the nutrients of the soil, storage of the water and avoid excessive depends on synthetic fertilizer and insecticides. In this case, eco- friendly cultivation could be a potential alternative solution to minimize effects of agriculture in our environment. The objective of this review paper is about organic cultivation following in particular biotechnological process focused on bio-fertilizer and bio-pesticides. Intense practice of chemical pesticides, insecticides has severe effect on both in human life and biodiversity. This cultivation process introduces farmer an alternative way which is nonhazardous, cost effective and ecofriendly. Organic fertilizer such as tea residue, ashes might be the best alternative to synthetic fertilizer those play important role in increasing soil nutrient and fertility. Ashes contain different essential and non-essential mineral contents that are required for plant growth. Organic pesticide such as neem spray is beneficial for crop as it is toxic for pest and insects. Recycled and composted crop wastes and animal manures, crop rotation, green manures and legumes etc. are suitable for soil fertility which is free from hazardous chemicals practice. Finally water hyacinth and algae are potential source of nutrients even alternative to soil for cultivation along with storage of water for continuous supply. Inorganic practice of agriculture, consuming fruits and vegetables becomes a threat for both human life and eco-system and synthetic fertilizer and pesticides are responsible for it. Farmers that practice eco-friendly farming have to implement steps to protect the environment, particularly by severely limiting the use of pesticides and avoiding the use of synthetic chemical fertilizers, which are necessary for organic systems to experience reduced environmental harm and health risk.

Keywords: organic farming, biopesticides, organic nutrients, water storage, global warming

Procedia PDF Downloads 60

Authors: Andrés Lara Guillén, José M. Soriano Disla, Gemma Castejón Martínez, David Fernández-Gutiérrez

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The worldwide population is exponentially increasing, which causes a rising demand for food, energy and non-renewable resources. These demands must be attended to from a circular economy point of view. Under this approach, the obtention of strategic products from biowaste is crucial for the society to keep the current lifestyle reducing the environmental and social issues linked to the lineal economy. This is the main objective of the VALUEWASTE project. VALUEWASTE is about valorizing urban biowaste into proteins for food and feed and biofertilizers, closing the loop of this waste stream. In order to achieve this objective, the project validates three value chains, which begin with the anaerobic digestion of the biowaste. From the anaerobic digestion, three by-products are obtained: i) methane that is used by microorganisms, which will be transformed into microbial proteins; ii) digestate that is used by black soldier fly, producing insect proteins; and iii) a nutrient-rich effluent, which will be transformed into biofertilizers. VALUEWASTE is an innovative solution, which combines different technologies to valorize entirely the biowaste. However, it is also required to demonstrate that the solution is greener than other traditional technologies (baseline systems). On one hand, the proteins from microorganisms and insects will be compared with other reference protein production systems (gluten, whey and soybean). On the other hand, the biofertilizers will be compared to the production of mineral fertilizers (ammonium sulphate and synthetic struvite). Therefore, the aim of this study is to provide that biowaste valorization can reduce the environmental impacts linked to both traditional proteins manufacturing processes and mineral fertilizers, not only at a pilot-scale but also at an industrial one. In the present study, both baseline system and VALUEWASTE solution are evaluated through the Environmental Life Cycle Assessment (E-LCA). The E-LCA is based on the standards ISO 14040 and 14044. The Environmental Footprint methodology was the one used in this study to evaluate the environmental impacts. The results for the baseline cases show that the food proteins coming from whey have the highest environmental impact on ecosystems compared to the other proteins sources: 7.5 and 15.9 folds higher than soybean and gluten, respectively. Comparing feed soybean and gluten, soybean has an environmental impact on human health 195.1 folds higher. In the case of biofertilizers, synthetic struvite has higher impacts than ammonium sulfate: 15.3 (ecosystems) and 11.8 (human health) fold, respectively. The results shown in the present study will be used as a reference to demonstrate the better environmental performance of the bio-based products obtained through the VALUEWASTE solution. Other originalities that the E-LCA performed in the VALUEWASTE project provides are the diverse direct implications on investment and policies. On one hand, better environmental performance will serve to remove the barriers linked to these kinds of technologies, boosting the investment that is backed by the E-LCA. On the other hand, it will be a germ to design new policies fostering these types of solutions to achieve two of the key targets of the European Community: being self-sustainable and carbon neutral.

Keywords: anaerobic digestion, biofertilizers, circular economy, nutrients recovery

Procedia PDF Downloads 88

Authors: Lucas M. Furlan, Cesar A. Moreira, Jepherson F. Sales, Guilherme T. Bueno, Manuel E. Ferreira, Carla V. S. Coelho, Vania Rosolen

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Located in the western part of the State of Minas Gerais, Brazil, the study area consists of a savanna environment, represented by sedimentary plateau and a soil cover composed by lateritic and hydromorphic soils - in the latter, occurring the deferruginization and concentration of high-alumina clays, exploited as refractory material. In the hydromorphic topographic depressions (wetlands) the hydropedogical relationships are little known, but it is observed that in times of rainfall, the depressed region behaves like a natural seasonal reservoir - which suggests that the wetlands on the surface of the plateau are places of recharge of the aquifer. The aquifer recharge areas are extremely important for the sustainable social, economic and environmental development of societies. The understanding of hydrodynamics in relation to the functioning of the ferruginous and hydromorphic lateritic soils system in the savanna environment is a subject rarely explored in the literature, especially its understanding through the joint application of geoprocessing by UAV (Unmanned Aerial Vehicle) and electrical tomography. The objective of this work is to understand the hydrogeological dynamics in a wetland (with an area of 426.064 m²), in the Brazilian savanna,as well as the understanding of the subsurface architecture of hydromorphic depressions in relation to the recharge of aquifers. The wetland was compartmentalized in three different regions, according to the geoprocessing. Hydraulic conductivity studies were performed in each of these three portions. Electrical tomography was performed on 9 lines of 80 meters in length and spaced 10 meters apart (direction N45), and a line with 80 meters perpendicular to all others. With the data, it was possible to generate a 3D cube. The integrated analysis showed that the area behaves like a natural seasonal reservoir in the months of greater precipitation (December – 289mm; January – 277,9mm; February – 213,2mm), because the hydraulic conductivity is very low in all areas. In the aerial images, geotag correction of the images was performed, that is, the correction of the coordinates of the images by means of the corrected coordinates of the Positioning by Precision Point of the Brazilian Institute of Geography and Statistics (IBGE-PPP). Later, the orthomosaic and the digital surface model (DSM) were generated, which with specific geoprocessing generated the volume of water that the wetland can contain - 780,922m³ in total, 265,205m³ in the region with intermediate flooding and 49,140m³ in the central region, where a greater accumulation of water was observed. Through the electrical tomography it was possible to identify that up to the depth of 6 meters the water infiltrates vertically in the central region. From the 8 meters depth, the water encounters a more resistive layer and the infiltration begins to occur horizontally - tending to concentrate the recharge of the aquifer to the northeast and southwest of the wetland. The hydrodynamics of the area is complex and has many challenges in its understanding. The next step is to relate hydrodynamics to the evolution of the landscape, with the enrichment of high-alumina clays, and to propose a management model for the seasonal reservoir.

Keywords: electrical tomography, hydropedology, unmanned aerial vehicle, water resources management

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Authors: Ayman Hijazi, Witold Kwapinski, J. J. Leahy

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Finding a sustainable alternative energy to fossil fuel is an urgent need as various environmental challenges in the world arise. Therefore, formic acid (FA) decomposition has been an attractive field that lies at the center of the biomass platform, comprising a potential pool of hydrogen energy that stands as a distinct energy vector. Liquid FA features considerable volumetric energy density of 6.4 MJ/L and a specific energy density of 5.3 MJ/Kg that qualifies it in the prime seat as an energy source for transportation infrastructure. Additionally, the increasing research interest in FA decomposition is driven by the need for in-situ H₂ production, which plays a key role in the hydrogenation reactions of biomass into higher-value components. It is reported elsewhere in the literature that catalytic decomposition of FA is usually performed in poorly designed setups using simple glassware under magnetic stirring, thus demanding further energy investment to retain the used catalyst. Our work suggests an approach that integrates designing a distinct catalyst featuring magnetic properties with a robust setup that minimizes experimental & measurement discrepancies. One of the most prominent active species for the dehydrogenation/hydrogenation of biomass compounds is palladium. Accordingly, we investigate the potential of engrafting palladium metal onto functionalized magnetic nanoparticles as a heterogeneous catalyst to favor the production of CO-free H₂ gas from FA. Using an ordinary magnet to collect the spent catalyst renders core-shell magnetic nanoparticles as the backbone of the process. Catalytic experiments were performed in a jacketed batch reactor equipped with an overhead stirrer under an inert medium. Through a distinct approach, FA is charged into the reactor via a high-pressure positive displacement pump at steady-state conditions. The produced gas (H₂+CO₂) was measured by connecting the gas outlet to a measuring system based on the amount of the displaced water. The uniqueness of this work lies in designing a very responsive catalyst, pumping a consistent amount of FA into a sealed reactor running at steady-state mild temperatures, and continuous gas measurement, along with collecting the used catalyst without the need for centrifugation. Catalyst characterization using TEM, XRD, SEM, and CHN elemental analyzer provided us with details of catalyst preparation and facilitated new venues to alter the nanostructure of the catalyst framework. Consequently, the introduction of amine groups has led to appreciable improvements in terms of dispersion of the doped metals and eventually attaining nearly complete conversion (100%) of FA after 7 hours. The relative importance of the process parameters such as temperature (35-85°C), stirring speed (150-450rpm), catalyst loading (50-200mgr.), and Pd doping ratio (0.75-1.80wt.%) on gas yield was assessed by a Taguchi design-of-experiment based model. Experimental results showed that operating at a lower temperature range (35-50°C) yielded more gas, while the catalyst loading and Pd doping wt.% were found to be the most significant factors with P-values 0.026 & 0.031, respectively.

Keywords: formic acid decomposition, green catalysis, hydrogen, mesoporous silica, process optimization, nanoparticles

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Authors: Vasundhara Arora, Anupam Jhobta, Suresh Thakur, Sanjiv Sharma

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Aims and Objectives: Intracranial tuberculosis (TB) is one of the most devastating manifestations of TB and a challenging public health issue of considerable importance and magnitude world over. This study elaborates on the imaging spectrum of neurotuberculosis on magnetic resonance imaging (MRI) in 29 clinically suspected cases from a tertiary care hospital. Materials and Methods: The prospective hospital based evaluation of MR imaging features of neuro-tuberculosis in 29 clinically suspected cases was carried out in Department of Radio-diagnosis, Indira Gandhi Medical Hospital from July 2017 to August 2018. MR Images were obtained on a 1.5 T Magnetom Avanto machine and were analyzed to identify any abnormal meningeal enhancement or parenchymal lesions. Microbiological and Biochemical CSF analysis was performed in radio-logically suspected cases and the results were compared with the imaging data. Clinical follow up of the patients started on anti-tuberculous treatment was done to evaluate the response to treatment and clinical outcome. Results: Age range of patients in the study was between 1 year to 73 years. The mean age of presentation was 11.5 years. No significant difference in the distribution of cerebral tuberculosis was noted among the two genders. Imaging findings of neuro-tuberculosis obtained were varied and non specific ranging from lepto-meningeal enhancement, cerebritis to space occupying lesions such as tuberculomas and tubercular abscesses. Complications presenting as hydrocephalus (n= 7) and infarcts (n=9) was noted in few of these patients. 29 patients showed radiological suspicion of CNS tuberculosis with meningitis alone observed in 11 cases, tuberculomas alone were observed in 4 cases, meningitis with parenchymal tuberculomas in 11 cases. Tubercular abscess and cerebritis were observed in one case each. Tuberculous arachnoiditis was noted in one patient. Gene expert positivity was obtained in 11 out of 29 radiologically suspected patients; none of the patients showed culture positivity. Meningeal form of the disease alone showed higher positivity rate of gene Xpert (n=5) followed by combination of meningeal and parenchymal forms of disease (n=4). The parenchymal manifestation of disease alone showed least positivity rates (n= 3) with gene xpert testing. All 29 patients were started on anti tubercular treatment based on radiological suspicion of the disease with clinical improvement observed in 27 treated patients. Conclusions: In our study, higher incidence of neuro- tuberculosis was noted in paediatric population with predominance of the meningeal form of the disease. Gene Xpert positivity obtained was low due to paucibacillary nature of cerebrospinal fluid (CSF) with even lower positivity of CSF samples in parenchymal form of the manifestation. MRI showed high accuracy in detecting CNS lesions in neuro-tuberculosis. Hence, it can be concluded that MRI plays a crucial role in the diagnosis because of its inherent sensitivity and specificity and is an indispensible imaging modality. It caters to the need of early diagnosis owing to poor sensitivity of microbiological tests more so in the parenchymal manifestation of the disease.

Keywords: neurotuberculosis, tubercular abscess, tuberculoma, tuberculous meningitis

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Authors: A. Augustyn, M. Florek, M. Herezniak

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Since the public authorities are constantly pressured by the public opinion to showcase the tangible and measurable results of their efforts, the evaluation of place brand-related activities becomes a necessity. Given the political and social character of place branding process, the legitimization of the branding efforts requires the compliance of the objectives set out in the city brand strategy with the actual needs, expectations, and aspirations of various internal stakeholders. To deliver on the diverse promises, city authorities and brand managers need to translate them into the measurable indicators against which the brand strategy effectiveness will be evaluated. In concert with these observations are the findings from branding and marketing literature with a widespread consensus that places should adopt a more systematic and holistic approach in order to ensure the performance of their brands. However, the measurement of the effectiveness of place branding remains insufficiently explored in theory, even though it is considered a significant step in the process of place brand management. Therefore, the aim of the research presented in the current paper was to collect insights on the nature of effectiveness measurement of city brand strategies and to juxtapose these findings with the theoretical assumptions formed on the basis of the state-of-the-art literature review. To this end, 15 international academic experts (out of 18 initially selected) with affiliation from ten countries (five continents), were individually interviewed. The standardized set of 19 open-ended questions was used for all the interviewees, who had been selected based on their expertise and reputation in the fields of place branding/marketing. Findings were categorized into four modules: (i) conceptualizations of city brand effectiveness, (ii) methodological issues of city brand effectiveness measurement, (iii) the nature of measurement process, (iv) articulation of key performance indicators (KPIs). Within each module, the interviewees offered diverse insights into the subject based on their academic expertise and professional activity as consultants. They proposed that there should be a twofold understanding of effectiveness. The narrow one when it is conceived as the aptitude to achieve specific goals, and the broad one in which city brand effectiveness is seen as an increase in social and economic reality of a place, which in turn poses diverse challenges for the measurement concepts and processes. Moreover, the respondents offered a variety of insights into the methodological issues, particularly about the need for customization and flexibility of the measurement systems, for the employment of interdisciplinary approach to measurement and implications resulting therefrom. Considerable emphasis was put on the inward approach to measurement, namely the necessity to monitor the resident’s evaluation of brand related activities instead of benchmarking cities against the competitive set. Other findings encompass the issues of developing appropriate KPIs for the city brand, managing the measurement process and the inclusion of diverse stakeholders to produce a sound measurement system. Furthermore, the interviewees enumerated the most frequently made mistakes in measurement mainly resulting from the misunderstanding of the nature of city brands. This research was financed by the National Science Centre, Poland, research project no. 2015/19/B/HS4/00380 Towards the categorization of place brand strategy effectiveness indicators – findings from strategic documents of Polish district cities – theoretical and empirical approach.

Keywords: city branding, effectiveness, experts’ insights, measurement

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Authors: Jean Felix Tuyisingize

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Speeding or inappropriate speed is a major problem worldwide, contributing to 10-15% of road crashes and 30% of fatal injury crashes. The consequences of speeding put the driver's life at risk and the lives of other road users like motorists, cyclists, and pedestrians. To control vehicle speeds, governments, and traffic authorities enforced speed regulations through speed cameras and speed section control, which monitor all vehicle speeds and detect plate numbers to levy penalties. However, speed limit violations are prevalent, even on motorways with speed cameras. The problem with speed cameras is that they alter driver behaviors, and their effect declines with increasing distance from the speed camera location. Drivers decelerate short distances before the camera and vigorously accelerate above the speed limit just after passing by the camera. The sudden decelerating near cameras causes the drivers to try to make up for lost time after passing it, and they do this by speeding up, resulting in a phenomenon known as the "Kangaroo jump" or "V-profile" around camera/ASSC areas. This study investigated the impact of speed enforcement devices, specifically Average Speed Section Control (ASSCs) and fixed cameras, on acceleration and deceleration events within their vicinity. The research employed advanced statistical and Geographic Information System (GIS) analysis on naturalistic driving data, to uncover speeding patterns near the speed enforcement systems. The study revealed a notable concentration of events within a 600-meter radius of enforcement devices, suggesting their influence on driver behaviors within a specific range. However, most of these events are of low severity, suggesting that drivers may not significantly alter their speed upon encountering these devices. This behavior could be attributed to several reasons, such as consistently maintaining safe speeds or using real-time in-vehicle intervention systems. The complexity of driver behavior is also highlighted, indicating the potential influence of factors like traffic density, road conditions, weather, time of day, and driver characteristics. Further, the study highlighted that high-severity events often occurred outside speed enforcement zones, particularly around intersections, indicating these as potential hotspots for drastic speed changes. These findings call for a broader perspective on traffic safety interventions beyond reliance on speed enforcement devices. However, the study acknowledges certain limitations, such as its reliance on a specific geographical focus, which may impact the broad applicability of the findings. Additionally, the severity of speed modification events was categorized into low, medium, and high, which could oversimplify the continuum of speed changes and potentially mask trends within each category. This research contributes valuable insights to traffic safety and driver behavior literature, illuminating the complexity of driver behavior and the potential influence of factors beyond the presence of speed enforcement devices. Future research directions may employ various categories of event severity. They may also explore the role of in-vehicle technologies, driver characteristics, and a broader set of environmental variables in driving behavior and traffic safety.

Keywords: acceleration, deceleration, speeding, inappropriate speed, speed enforcement cameras

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

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

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

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Authors: Sonika Sharma, Mohan G. Vairale, Sibnarayan Datta, Soumya Chatterjee, Dharmendra Dubey, Rajesh Prasad, Raghvendra Budhauliya, Bidisha Das, Vijay Veer

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Bacteriophages are viruses that parasitize specific bacteria and multiply in metabolising host bacteria. Bacteriophages hunt for a single or a subset of bacterial species, making them potential antibacterial agents. Utilizing the ability of phages to control bacterial populations has several applications from medical to the fields of agriculture, aquaculture and the food industry. However, harnessing phage based techniques in wastewater treatments to improve quality of effluent and sludge release into the environment is a potential area for R&D application. Phage mediated bactericidal effect in any wastewater treatment process has many controlling factors that lead to treatment performance. In laboratory conditions, titer of bacteriophages (coliphages) isolated from effluent water of a specially designed anaerobic digester of human night soil (DRDO Biotoilet) was successfully increased with a modified protocol of the classical double layer agar technique. Enrichment of the same was carried out and efficacy of the phage enriched medium was evaluated at different conditions (specific media, temperature, storage conditions). Growth optimization study was carried out on different media like soybean casein digest medium (Tryptone soya medium), Luria-Bertani medium, phage deca broth medium and MNA medium (Modified nutrient medium). Further, temperature-phage yield relationship was also observed at three different temperatures 27˚C, 37˚C and 44˚C at laboratory condition. Results showed the higher activity of coliphage 27˚C and at 37˚C. Further, addition of divalent ions (10mM MgCl2, 5mM CaCl2) and 5% glycerol resulted in a significant increase in phage titer. Besides this, effect of antibiotics addition like ampicillin and kanamycin at different concentration on plaque formation was analysed and reported that ampicillin at a concentration of 1mg/ml ampicillin stimulates phage infection and results in more number of plaques. Experiments to test viability of phage showed that it can remain active for 6 months at 4˚C in fresh tryptone soya broth supplemented with fresh culture of coliforms (early log phase). The application of bacteriophages (especially coliphages) for treatment of effluent of human faecal matter contaminated effluent water is unique. This environment-friendly treatment system not only reduces the pathogenic coliforms, but also decreases the competition between nuisance bacteria and functionally important microbial populations. Therefore, the phage based cocktail to treat fecal pathogenic bacteria present in black water has many implication in wastewater treatment processes including ‘DRDO Biotoilet’, which is an ecofriendly appropriate and affordable human faecal matter treatment technology for different climates and situations.

Keywords: wastewater, microbes, virus, biotoilet, phage viability

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Authors: Antonio Boggia, Luisa Paolotti, Gianluca Massei, Lucia Rocchi, Elaine Pace, Maria Attard

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The aim of this paper is to present a methodology and a computer model for sustainability assessment based on the integration of Multi-criteria Decision Analysis (MCDA) with a Geographic Information System (GIS). It presents the result of a study for the implementation of a model for measuring sustainability to address the policy actions for the improvement of sustainability at territory level. The aim is to rank areas in order to understand the specific technical and/or financial support that is required to develop sustainable growth. Assessing sustainable development is a multidimensional problem: economic, social and environmental aspects have to be taken into account at the same time. The tool for a multidimensional representation is a proper set of indicators. The set of indicators must be integrated into a model, that is an assessment methodology, to be used for measuring sustainability. The model, developed by the Environmental Laboratory of the University of Perugia, is called GeoUmbriaSUIT. It is a calculation procedure developed as a plugin working in the open-source GIS software QuantumGIS. The multi-criteria method used within GeoUmbriaSUIT is the algorithm TOPSIS (Technique for Order Preference by Similarity to Ideal Design), which defines a ranking based on the distance from the worst point and the closeness to an ideal point, for each of the criteria used. For the sustainability assessment procedure, GeoUmbriaSUIT uses a geographic vector file where the graphic data represent the study area and the single evaluation units within it (the alternatives, e.g. the regions of a country, or the municipalities of a region), while the alphanumeric data (attribute table), describe the environmental, economic and social aspects related to the evaluation units by means of a set of indicators (criteria). The use of the algorithm available in the plugin allows to treat individually the indicators representing the three dimensions of sustainability, and to compute three different indices: environmental index, economic index and social index. The graphic output of the model allows for an integrated assessment of the three dimensions, avoiding aggregation. The presence of separate indices and graphic output make GeoUmbriaSUIT a readable and transparent tool, since it doesn’t produce an aggregate index of sustainability as final result of the calculations, which is often cryptic and difficult to interpret. In addition, it is possible to develop a “back analysis”, able to explain the positions obtained by the alternatives in the ranking, based on the criteria used. The case study presented is an assessment of the level of sustainability in the six regions of Malta, an island state in the middle of the Mediterranean Sea and the southernmost member of the European Union. The results show that the integration of MCDA-GIS is an adequate approach for sustainability assessment. In particular, the implemented model is able to provide easy to understand results. This is a very important condition for a sound decision support tool, since most of the time decision makers are not experts and need understandable output. In addition, the evaluation path is traceable and transparent.

Keywords: GIS, multi-criteria analysis, sustainability assessment, sustainable development

Procedia PDF Downloads 289

Authors: Dan Kanmegne

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Africa will become the most populated continent by the end of the century, with around 4 billion inhabitants. Food security and climate changes will become continental issues since agricultural practices depend on climate but also contribute to global emissions and land degradation. Agroforestry has been identified as a cost-efficient and reliable strategy to address these two issues. It is defined as the integrated management of trees and crops/animals in the same land unit. Agroforestry provides benefits in terms of goods (fruits, medicine, wood, etc.) and services (windbreaks, fertility, etc.), and is acknowledged to have a great potential for carbon sequestration; therefore it can be integrated into reduction mechanisms of carbon emissions. Particularly in sub-Saharan Africa, the constraint stands in the lack of information about both areas under agroforestry and the characterization (composition, structure, and management) of each agroforestry system at the country level. This study describes and quantifies “what is where?”, earliest to the quantification of carbon stock in different systems. Remote sensing (RS) is the most efficient approach to map such a dynamic technology as agroforestry since it gives relatively adequate and consistent information over a large area at nearly no cost. RS data fulfill the good practice guidelines of the Intergovernmental Panel On Climate Change (IPCC) that is to be used in carbon estimation. Satellite data are getting more and more accessible, and the archives are growing exponentially. To retrieve useful information to support decision-making out of this large amount of data, satellite data needs to be organized so to ensure fast processing, quick accessibility, and ease of use. A new solution is a data cube, which can be understood as a multi-dimensional stack (space, time, data type) of spatially aligned pixels and used for efficient access and analysis. A data cube for Burkina Faso has been set up from the cooperation project between the international service provider WASCAL and Germany, which provides an accessible exploitation architecture of multi-temporal satellite data. The aim of this study is to map and characterize agroforestry systems using the Burkina Faso earth observation data cube. The approach in its initial stage is based on an unsupervised image classification of a normalized difference vegetation index (NDVI) time series from 2010 to 2018, to stratify the country based on the vegetation. Fifteen strata were identified, and four samples per location were randomly assigned to define the sampling units. For safety reasons, the northern part will not be part of the fieldwork. A total of 52 locations will be visited by the end of the dry season in February-March 2020. The field campaigns will consist of identifying and describing different agroforestry systems and qualitative interviews. A multi-temporal supervised image classification will be done with a random forest algorithm, and the field data will be used for both training the algorithm and accuracy assessment. The expected outputs are (i) map(s) of agroforestry dynamics, (ii) characteristics of different systems (main species, management, area, etc.); (iii) assessment report of Burkina Faso data cube.

Keywords: agroforestry systems, Burkina Faso, earth observation data cube, multi-temporal image classification

Procedia PDF Downloads 145

Authors: Reggie Chandra

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Trafficaccidents and traffic signal optimization are correlated. However, 70-90% of the traffic signals across the USA are not synchronized. The reason behind that is insufficient resources to create and implement timing plans. In this work, we will discuss the use of a breakthrough Artificial Intelligence (AI) technology to optimize traffic flow and collect 24/7/365 accurate traffic data using a vehicle detection system. We will discuss what are recent advances in Artificial Intelligence technology, how does AI work in vehicles, pedestrians, and bike data collection, creating timing plans, and what is the best workflow for that. Apart from that, this paper will showcase how Artificial Intelligence makes signal timing affordable. We will introduce a technology that uses Convolutional Neural Networks (CNN) and deep learning algorithms to detect, collect data, develop timing plans and deploy them in the field. Convolutional Neural Networks are a class of deep learning networks inspired by the biological processes in the visual cortex. A neural net is modeled after the human brain. It consists of millions of densely connected processing nodes. It is a form of machine learning where the neural net learns to recognize vehicles through training - which is called Deep Learning. The well-trained algorithm overcomes most of the issues faced by other detection methods and provides nearly 100% traffic data accuracy. Through this continuous learning-based method, we can constantly update traffic patterns, generate an unlimited number of timing plans and thus improve vehicle flow. Convolutional Neural Networks not only outperform other detection algorithms but also, in cases such as classifying objects into fine-grained categories, outperform humans. Safety is of primary importance to traffic professionals, but they don't have the studies or data to support their decisions. Currently, one-third of transportation agencies do not collect pedestrian and bike data. We will discuss how the use of Artificial Intelligence for data collection can help reduce pedestrian fatalities and enhance the safety of all vulnerable road users. Moreover, it provides traffic engineers with tools that allow them to unleash their potential, instead of dealing with constant complaints, a snapshot of limited handpicked data, dealing with multiple systems requiring additional work for adaptation. The methodologies used and proposed in the research contain a camera model identification method based on deep Convolutional Neural Networks. The proposed application was evaluated on our data sets acquired through a variety of daily real-world road conditions and compared with the performance of the commonly used methods requiring data collection by counting, evaluating, and adapting it, and running it through well-established algorithms, and then deploying it to the field. This work explores themes such as how technologies powered by Artificial Intelligence can benefit your community and how to translate the complex and often overwhelming benefits into a language accessible to elected officials, community leaders, and the public. Exploring such topics empowers citizens with insider knowledge about the potential of better traffic technology to save lives and improve communities. The synergies that Artificial Intelligence brings to traffic signal control and data collection are unsurpassed.

Keywords: artificial intelligence, convolutional neural networks, data collection, signal control, traffic signal

Procedia PDF Downloads 169

Authors: Lazzat M. Zhamaliyeva, Nurgul A. Abenova, Gauhar S. Dilmagambetova, Ziyash Zh. Tanbetova, Moldir B. Ahmetzhanova, Tatyana P. Ostretcova, Aliya A. Yegemberdiyeva

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Introduction: There are many reasons for the weak training of family doctors in Kazakhstan: the unified national educational program is not focused on competencies, the role of a general practitioner (GP) is not clear, poor funding for the health care and education system, outdated teaching and assessment methods, inefficient management. We highlight two issues in particular. Firstly, academic teachers of family medicine (FM) in Kazakhstan do not practice as family doctors; most of them are narrow specialists (pediatricians, therapists, surgeons, etc.); they usually hold one-time consultations; clinical mentors from practical healthcare (non-academic teachers) do not have the teaching competences, and the vast majority of them are also narrow specialists. Secondly, clinical sites (polyclinics) are unprepared for general practice and do not follow the principles of family medicine; residents do not like to be in primary health care (PHC) settings due to the chaos that is happening there, as well as due to the lack of the necessary equipment for mastering and consolidating practical skills. Aim: We present the concept of the family physicians’ training office (FPTO), which is being created as a friendly learning environment for young general practitioners and for the involvement of academic teachers of family medicine in the practical work and innovative development of PHC. Methodology: In developing the conceptual framework and identifying practical activities, we drew on literature and expert input, and interviews. Results: The goal of the FPTO is to create a favorable educational and clinical environment for the development of the FM residents’ competencies, in which the residents with academic teachers and clinical mentors could understand and accept the principles of family medicine, improve clinical knowledge and skills, and gain experience in improving the quality of their practice in scientific basis. Three main areas of office activity are providing primary care to the patients, improving educational services for FM residents and other medical workers, and promoting research in PHC and innovations. The office arranges for residents to see outpatients at least 50% of the time, and teachers of FM departments at least 1/4 of their working time conduct general medical appointments next to residents. Taking into account the educational and scientific workload, the number of attached population for one GP does not exceed 500 persons. The equipment of the office allows FPTO workers to perform invasive and other manipulations without being sent to other clinics. In the office, training for residents is focused on their needs and aimed at achieving the required level of competence. International methodologies and assessment tools are adapted to local conditions and evaluated for their effectiveness and acceptability. Residents and their faculty actively conduct research in the field of family medicine. Conclusions: We propose to change the learning environment in order to create teams of like-minded people, to unite residents and teachers even more for the development of family medicine. The offices will also invest resources in developing and maintaining young doctors' interest in family medicine.

Keywords: educational environment, family medicine residents, family physicians’ training office, primary care research

Procedia PDF Downloads 134

Authors: Pierobon Enrico, Missiaggia Marta, Castelluzzo Michele, Tommasino Francesco, Ricci Leonardo, Scifoni Emanuele, Vincezo Monaco, Boscardin Maurizio, La Tessa Chiara

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Clinical outcomes collected over the past three decades have suggested that ion therapy has the potential to be a treatment modality superior to conventional radiation for several types of cancer, including recurrences, as well as for other diseases. Although the results have been encouraging, numerous treatment uncertainties remain a major obstacle to the full exploitation of particle radiotherapy. To overcome therapy uncertainties optimizing treatment outcome, the best possible radiation quality description is of paramount importance linking radiation physical dose to biological effects. Microdosimetry was developed as a tool to improve the description of radiation quality. By recording the energy deposition at the micrometric scale (the typical size of a cell nucleus), this approach takes into account the non-deterministic nature of atomic and nuclear processes and creates a direct link between the dose deposited by radiation and the biological effect induced. Microdosimeters measure the spectrum of lineal energy y, defined as the energy deposition in the detector divided by most probable track length travelled by radiation. The latter is provided by the so-called “Mean Chord Length” (MCL) approximation, and it is related to the detector geometry. To improve the characterization of the radiation field quality, we define a new quantity replacing the MCL with the actual particle track length inside the microdosimeter. In order to measure this new quantity, we propose a two-stage detector consisting of a commercial Tissue Equivalent Proportional Counter (TEPC) and 4 layers of Low Gain Avalanche Detectors (LGADs) strips. The TEPC detector records the energy deposition in a region equivalent to 2 um of tissue, while the LGADs are very suitable for particle tracking because of the thickness thinnable down to tens of micrometers and fast response to ionizing radiation. The concept of HDM has been investigated and validated with Monte Carlo simulations. Currently, a dedicated readout is under development. This two stages detector will require two different systems to join complementary information for each event: energy deposition in the TEPC and respective track length recorded by LGADs tracker. This challenge is being addressed by implementing SoC (System on Chip) technology, relying on Field Programmable Gated Arrays (FPGAs) based on the Zynq architecture. TEPC readout consists of three different signal amplification legs and is carried out thanks to 3 ADCs mounted on a FPGA board. LGADs activated strip signal is processed thanks to dedicated chips, and finally, the activated strip is stored relying again on FPGA-based solutions. In this work, we will provide a detailed description of HDM geometry and the SoC solutions that we are implementing for the readout.

Keywords: particle tracking, ion therapy, low gain avalanche diode, tissue equivalent proportional counter, microdosimetry

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Authors: Mark McGuire, Eric Shannon, John Parmigiani

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Timber harvesting increasingly involves mechanized equipment. This has increased the efficiency of harvesting, but has also introduced worker-safety concerns. One such concern arises from the use of harvesters. During operation, harvesters subject saw chain to large dynamic mechanical stresses. These stresses can, under certain conditions, cause the saw chain to fracture. The high speed of harvester saw chain can cause the resulting open chain loop to fracture a second time due to the dynamic loads placed upon it as it travels through space. If a second fracture occurs, it can result in a projectile consisting of one-to-several chain links. This projectile is referred to as a chain shot. It has speeds similar to a bullet but typically has greater mass and is a significant safety concern. Numerous examples exist of chain shots penetrating bullet-proof barriers and causing severe injury and death. Improved harvester-cab barriers can help prevent injury however a comprehensive scientific understanding of chain shot is required to consistently reduce or prevent it. Obtaining this understanding requires a test machine with the capability to cause chain shot to occur under carefully controlled conditions and accurately measure the response. Worldwide few such test machine exist. Those that do focus on validating the ability of barriers to withstand a chain shot impact rather than obtaining a scientific understanding of the chain shot event itself. The purpose of this paper is to describe the design, fabrication, and use of a test machine capable of a comprehensive scientific investigation of chain shot. The capabilities of this machine are to test all commercially-available saw chains and bars at chain tensions and speeds meeting and exceeding those typically encountered in harvester use and accurately measure the corresponding key technical parameters. The test machine was constructed inside of a standard shipping container. This provides space for both an operator station and a test chamber. In order to contain the chain shot under any possible test conditions, the test chamber was lined with a base layer of AR500 steel followed by an overlay of HDPE. To accommodate varying bar orientations and fracture-initiation sites, the entire saw chain drive unit and bar mounting system is modular and capable of being located anywhere in the test chamber. The drive unit consists of a high-speed electric motor with a flywheel. Standard Ponsse harvester head components are used to bar mounting and chain tensioning. Chain lubrication is provided by a separate peristaltic pump. Chain fracture is initiated through ISO standard 11837. Measure parameters include shaft speed, motor vibration, bearing temperatures, motor temperature, motor current draw, hydraulic fluid pressure, chain force at fracture, and high-speed camera images. Results show that the machine is capable of consistently causing chain shot. Measurement output shows fracture location and the force associated with fracture as a function of saw chain speed and tension. Use of this machine will result in a scientific understanding of chain shot and consequently improved products and greater harvester operator safety.

Keywords: chain shot, safety, testing, timber harvesters

Procedia PDF Downloads 152

Authors: Irene T. Ho

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Students with Autism Spectrum Disorder (ASD) studying in mainstream schools often face difficulties adjusting to school life and teachers often find it challenging to meet the needs of these students. The Hong Kong Jockey Club Autism Support Network (JC A-Connect) is an initiative launched in 2015 to enhance support for students with ASD as well as their families and schools. The School Support Programme of the Project aims at building the capacity of schools to provide quality education for these students. The present report provides a summary of the main features of the support model and the related evaluation results. The school support model was conceptualized in response to four observed needs: (1) inadequate teacher expertise in dealing with the related challenges, (2) the need to promote evidence-based practices in schools, (3) less than satisfactory home-school collaboration and whole-school participation, and (4) lack of concerted effort by different parties involved in providing support to schools. The resulting model had partnership and capacity building as two guiding tenets for the School Support Programme. There were two levels of partnership promoted in the project. At the programme support level, a platform that enables effective collaboration among major stakeholders was established, including the funding body that provides the necessary resources, the Education Bureau that helps to engage schools, university experts who provide professional leadership and research support, as well as non-governmental organization (NGO) professionals who provide services to the schools. At the programme implementation level, tripartite collaboration among teachers, parents and professionals was emphasized. This notion of partnership permeated efforts at capacity building targeting students with ASD, school personnel, parents and peers. During 2015 to 2018, school-based programmes were implemented in over 400 primary and secondary schools with the following features: (1) spiral Tier 2 (group) training for students with ASD to enhance their adaptive skills, led by professionals but with strong teacher involvement to promote transfer of knowledge and skills; (2) supplementary programmes for teachers, parents and peers to enhance their capability to support students with ASD; and (3) efforts at promoting continuing or transfer of learning, on the part of both students and teachers, to Tier 1 (classroom practice) and Tier 3 (individual training) contexts. Over 5,000 students participated in the Programme, representing about 50% of students diagnosed with ASD in mainstream public sector schools in Hong Kong. Results showed that the Programme was effective in helping students improve to various extents at three levels: achievement of specific training goals, improvement in adaptive skills in school, and change in ASD symptoms. The sense of competence of teachers and parents in dealing with ASD-related issues, measured by self-report rating scales, was also significantly enhanced. Moreover, effects on enhancing the school system to provide support for students with ASD, assessed according to indicators of inclusive education, were seen. The process and results of this Programme illustrate how obstacles to inclusive education for students with ASD could be overcome by strengthening the necessary partnerships and building the required capabilities of all parties concerned.

Keywords: autism, school support, skills training, teacher development, three-tier model

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Authors: Xu Qian

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This abstract examines the concept of resilience in adolescents from both adolescent medicine and child psychology perspectives. It discusses the role of healthcare providers in fostering resilience among adolescents, encompassing physical, psychological, and social aspects. The paper highlights evidence-based interventions and practical strategies for promoting resilience in this population. Introduction: Resilience plays a crucial role in the healthy development of adolescents, enabling them to navigate through the challenges of this transitional period. This abstract explores the concept of resilience from the perspectives of adolescent medicine and child psychology, shedding light on the collective efforts of healthcare providers in fostering resilience. By integrating the principles and practices of these two disciplines, this abstract emphasizes the multidimensional nature of resilience and its significance in the overall well-being of adolescents. Methods: A comprehensive literature review was conducted, encompassing research articles, empirical studies, and expert opinions from both adolescent medicine and child psychology fields. The search included databases such as PubMed, PsycINFO, and Google Scholar, focusing on publications from the past decade. The review aimed to identify evidence-based interventions and practical strategies employed by healthcare providers to promote resilience among adolescents. Results: The review revealed several key findings regarding the promotion of resilience in adolescents. Firstly, resilience is a dynamic process influenced by individual characteristics, environmental factors, and the interaction between the two. Secondly, healthcare providers play a critical role in fostering resilience by addressing the physical, psychological, and social needs of adolescents. This entails comprehensive healthcare services that integrate medical care, mental health support, and social interventions. Thirdly, evidence-based interventions such as cognitive-behavioral therapy, social skills training, and positive youth development programs have shown promising outcomes in enhancing resilience. Discussion: The integration of adolescent medicine and child psychology perspectives provides a comprehensive framework for promoting resilience in adolescents. By acknowledging the interplay between physical health, psychological well-being, and social functioning, healthcare providers can tailor interventions to address the specific needs and challenges faced by adolescents. Collaborative efforts between medical professionals, psychologists, educators, and families are vital in creating a supportive environment that fosters resilience. Additionally, the findings highlight the importance of early identification and intervention, emphasizing the need for routine screening and assessment to identify adolescents at risk and provide timely support. Conclusion: Promoting resilience in adolescents requires a holistic approach that integrates adolescent medicine and child psychology perspectives. By recognizing the multifaceted nature of resilience, healthcare providers can implement evidence-based interventions and practical strategies to enhance the well-being of adolescents. The collaboration between healthcare professionals from different disciplines, alongside the involvement of families and communities, is crucial for creating a resilient support system. By investing in the promotion of resilience during adolescence, we can empower young individuals to overcome adversity and thrive in their journey toward adulthood.

Keywords: psychology, clinical psychology, child psychology, adolescent psychology, adolescent

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Authors: Francesco Pandolfi, Georgios Baltzopoulos, Iunio Iervolino

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The impact of extreme winds on industrial assets and the built environment is gaining increasing attention from stakeholders, including the corporate insurance industry. This has led to a progressively more in-depth study of building vulnerability and fragility to wind. Wind vulnerability models are used in probabilistic risk assessment to relate a loss metric to an intensity measure of the natural event, usually a gust or a mean wind speed. In fact, vulnerability models can be integrated with the wind hazard, which consists of associating a probability to each intensity level in a time interval (e.g., by means of return periods) to provide an assessment of future losses due to extreme wind. This has also given impulse to the world- and regional-scale wind hazard studies.Another approach often adopted for the probabilistic description of building vulnerability to the wind is the use of fragility functions, which provide the conditional probability that selected building components will exceed certain damage states, given wind intensity. In fact, in wind engineering literature, it is more common to find structural system- or component-level fragility functions rather than wind vulnerability models for an entire building. Loss assessment based on component fragilities requires some logical combination rules that define the building’s damage state given the damage state of each component and the availability of a consequence model that provides the losses associated with each damage state. When risk calculations are based on numerical simulation of a structure’s behavior during extreme wind scenarios, the interaction of component fragilities is intertwined with the computational procedure. However, simulation-based approaches are usually computationally demanding and case-specific. In this context, the present work introduces the ExtReMe wind risk assESsment prototype Software, ERMESS, which is being developed at the University of Naples Federico II. ERMESS is a wind risk assessment tool for insurance applications to industrial facilities, collecting a wide assortment of available wind vulnerability models and fragility functions to facilitate their incorporation into risk calculations based on in-built or user-defined wind hazard data. This software implements an alternative method for building-specific risk assessment based on existing component-level fragility functions and on a number of simplifying assumptions for their interactions. The applicability of this alternative procedure is explored by means of an illustrative proof-of-concept example, which considers four main building components, namely: the roof covering, roof structure, envelope wall and envelope openings. The application shows that, despite the simplifying assumptions, the procedure can yield risk evaluations that are comparable to those obtained via more rigorous building-level simulation-based methods, at least in the considered example. The advantage of this approach is shown to lie in the fact that a database of building component fragility curves can be put to use for the development of new wind vulnerability models to cover building typologies not yet adequately covered by existing works and whose rigorous development is usually beyond the budget of portfolio-related industrial applications.

Keywords: component wind fragility, probabilistic risk assessment, vulnerability model, wind-induced losses

Procedia PDF Downloads 181

Authors: Jenson George, Thoa Nguyen, Garth Sanewski, Craig Hardner, Heather Eunice Smyth

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Pineapple (Ananas comosus), with its unique sweet flavour, is one of the most popular tropical, non-climacteric fruits consumed worldwide. It is also the third most important tropical fruit in world production. In Australia, 99% of the pineapple production is from the Queensland state due to the favourable subtropical climatic conditions. The flavourful fruit is known to contain around 500 volatile organic compounds (VOC) at varying concentrations and greatly contribute to the flavour quality of pineapple fruit by providing distinct aroma sensory properties that are sweet, fruity, tropical, pineapple-like, caramel-like, coconut-like, etc. The aroma of pineapple is one of the important factors attracting consumers and strengthening the marketplace. To better understand the aroma of Australian-grown pineapples, the matrix-matched Gas chromatography–mass spectrometry (GC-MS), Head Space - Solid-phase microextraction (HS-SPME), Stable-isotope dilution analysis (SIDA) method was developed and validated. The developed method represents a significant improvement over current methods with the incorporation of multiple external reference standards, multiple isotopes labeled internal standards, and a matching model system of pineapple fruit matrix. This method was employed to quantify 28 key aroma compounds in more than 200 genetically diverse pineapple varieties from a breeding program. The Australian pineapple cultivars varied in content and composition of free volatile compounds, which were predominantly comprised of esters, followed by terpenes, alcohols, aldehydes, and ketones. Using selected commercial cultivars grown in Australia, and by employing the sensorial analysis, the appearance (colour), aroma (intensity, sweet, vinegar/tang, tropical fruits, floral, coconut, green, metallic, vegetal, fresh, peppery, fermented, eggy/sulphurous) and texture (crunchiness, fibrousness, and juiciness) were obtained. Relationships between sensory descriptors and volatiles were explored by applying multivariate analysis (PCA) to the sensorial and chemical data. The key aroma compounds of pineapple exhibited a positive correlation with corresponding sensory properties. The sensory and volatile data were also used to explore genetic diversity in the breeding population. GWAS was employed to unravel the genetic control of the pineapple volatilome and its interplay with fruit sensory characteristics. This study enhances our understanding of pineapple aroma (flavour) compounds, their biosynthetic pathways and expands breeding option for pineapple cultivars. This research provides foundational knowledge to support breeding programs, post-harvest and target market studies, and efforts to optimise the flavour of commercial pineapple varieties and their parent lines to produce better tasting fruits for consumers.

Keywords: Ananas comosus, pineapple, flavour, volatile organic compounds, aroma, Gas chromatography–mass spectrometry (GC-MS), Head Space - Solid-phase microextraction (HS-SPME), Stable-isotope dilution analysis (SIDA).

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Authors: Mirain Rhys, Kevin Smith

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In 2017, the Welsh Government introduced an ambitious, new strategy to increase the number of Welsh speakers in Wales to 1 million by 2050. The Welsh education system is a vitally important feature of this strategy. All children attending state schools in Wales learn Welsh as a second language until the age of 16 and are assessed at General Certificate of Secondary Education (GCSE) level. In 2013, a review of Welsh second language instruction in Key Stages 3 and 4 was completed. The report identified considerable gaps in teachers’ preparation and training for teaching Welsh; poor Welsh language ethos at many schools; and a general lack of resources to support the instruction of Welsh. Recommendations were made across a number of dimensions including curriculum content, pedagogical practice, and teacher assessment, training, and resources. With a new national curriculum currently in development, this study builds on this review and provides unprecedented detail into pupils’ and teachers’ perceptions of Welsh language instruction. The current research built on data taken from an existing capacity building research project on Welsh education, the Wales multi-cohort study (WMS). Quantitative data taken from WMS surveys with over 1200 pupils in schools in Wales indicated that Welsh language lessons were the least enjoyable subject among pupils. The current research aimed to unpick pupil experiences in order to add to the policy development context. To achieve this, forty-four pupils and four teachers in three schools from the larger WMS sample participated in focus groups. Participants from years 9, 11 and 13 who had indicated positive, negative and neutral attitudes towards the Welsh language in a previous WMS survey were selected. Questions were based on previous research exploring issues including, but not limited to pedagogy, policy, assessment, engagement and (teacher) training. A thematic analysis of the focus group recordings revealed that the majority of participants held positive views around keeping the language alive but did not want to take on responsibility for its maintenance. These views were almost entirely based on their experiences of learning Welsh at school, especially in relation to their perceived lack of choice and opinions around particular lesson strategies and assessment. Analysis of teacher interviews highlighted a distinct lack of resources (materials and staff alike) compared to modern foreign languages, which had a negative impact on student motivation and attitudes. Both staff and students indicated a need for more practical, oral language instruction which could lead to Welsh being used outside the classroom. The data corroborate many of the review’s previous findings, but what makes this research distinctive is the way in which pupils poignantly address generally misguided aims for Welsh language instruction, poor pedagogical practice and a general disconnect between Welsh instruction and its daily use in their lives. These findings emphasize the complexity of incorporating the educational sector in strategies for Welsh language maintenance and the complications arising from pedagogical training, support, and resources, as well as teacher and pupil perceptions of, and attitudes towards, teaching and learning Welsh.

Keywords: bilingual education, language maintenance, language revitalisation, minority languages, Wales

Procedia PDF Downloads 112

Authors: V. K. Tewari, M. Hussain, H. K. D. Gupta

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Background: Nitric Oxide Donars (NODs) (intrathecal sodium nitroprusside (ITSNP) and oral tadalafil 20mg post ITSNP) has been studied in this context in cerebral palsy patients for fast recovery. This work proposes two mechanisms for acute cases and one mechanism for chronic cases, which are interrelated, for physiological recovery. a) Retrograde Neurotransmission (acute cases): 1) Normal excitatory impulse: at the synaptic level, glutamate activates NMDA receptors, with nitric oxide synthetase (NOS) on the postsynaptic membrane, for further propagation by the calcium-calmodulin complex. Nitric oxide (NO, produced by NOS) travels backward across the chemical synapse and binds the axon-terminal NO receptor/sGC of a presynaptic neuron, regulating anterograde neurotransmission (ANT) via retrograde neurotransmission (RNT). Heme is the ligand-binding site of the NO receptor/sGC. Heme exhibits > 10,000-fold higher affinity for NO than for oxygen (the 10,000-fold effect) and is completed in 20 msec. 2) Pathological conditions: normal synaptic activity, including both ANT and RNT, is absent. A NO donor (SNP) releases NO from NOS in the postsynaptic region. NO travels backward across a chemical synapse to bind to the heme of a NO receptor in the axon terminal of a presynaptic neuron, generating an impulse, as under normal conditions. b) Vasopasm: (acute cases) Perforators show vasospastic activity. NO vasodilates the perforators via the NO-cAMP pathway. c) Long-Term Potentiation (LTP): (chronic cases) The NO–cGMP-pathway plays a role in LTP at many synapses throughout the CNS and at the neuromuscular junction. LTP has been reviewed both generally and with respect to brain regions specific for memory/learning. Aims/Study Design: The principles of “generation of impulses from the presynaptic region to the postsynaptic region by very potent RNT (10,000-fold effect)” and “vasodilation of arteriolar perforators” are the basis of the authors’ hypothesis to treat cerebral palsy cases. Case-control prospective study. Materials and Methods: The experimental population included 82 cerebral palsy patients (10 patients were given control treatments without NOD or with 5% dextrose superfusion, and 72 patients comprised the NOD group). The mean time for superfusion was 5 months post-cerebral palsy. Pre- and post-NOD status was monitored by Gross Motor Function Classification System for Cerebral Palsy (GMFCS), MRI, and TCD studies. Results: After 7 days in the NOD group, the mean change in the GMFCS score was an increase of 1.2 points mean; after 3 months, there was an increase of 3.4 points mean, compared to the control-group increase of 0.1 points at 3 months. MRI and TCD documented the improvements. Conclusions: NOD (ITSNP boosts up the recovery and oral tadalafil maintains the recovery to a well-desired level) acts swiftly in the treatment of CP, acting within 7 days on 5 months post-cerebral palsy either of the three mechanisms.

Keywords: cerebral palsy, intrathecal sodium nitroprusside, oral tadalafil, perforators, vasodilations, retrograde transmission, the 10, 000-fold effect, long-term potantiation

Procedia PDF Downloads 362

Authors: Charlotte Jeavons, Charitini Stavropoulous, Nicolas Drey

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Background: Over one-third of five-year-olds and almost half of all eight-year-olds in the UK have obvious caries experience that can be detected by visual screening techniques. School-based caries preventions programs to apply fluoride varnish to young children’s teeth operate in many areas in the UK. Their aim is to reduce dental caries in children. The Department of Health guidance (2009) on consent states information must be provided to parents to enable informed autonomous decision-making prior to any treatment involving their young children. Fluoride varnish schemes delivered in primary schools use letters for this purpose. Parents are expected to return these indicating their consent or refusal. A large proportion of parents do not respond. In the absence of positive consent, these children are excluded from the program. Non-response is more common in deprived areas creating inequality. The reason for this is unknown. The consent process used is underpinned by the ethical theory of deontology that is prevalent in clinical dentistry and widely accepted in bio-ethics. Objective: To investigate parents’ views, understanding and experience of the fluoride varnish program taking place in their child’s school, including their views about the practical consent arrangements. Method: Schools participating in the fluoride varnish scheme operating in Enfield, North London, were asked to take part. Parents with children in nursery, reception, or year one were invited to participate via semi-structured interviews and focus groups. Thematic analysis was conducted. Findings: 40 parents were recruited from eight schools. The global theme of ‘trust’ was identified as the strongest influence on parental responses. Six themes were identified; protecting children from harm is viewed by parents as their role, parents have the capability to decide but lack confidence, sharing responsibility for their child’s oral health with the State is welcomed by a parent, existing relationships within parents’ social networks strongly influences consent decisions, official dental information is not communicated effectively, sending a letter to parents’ and excluding them from meeting dental practitioners is ineffective. The information delivered via a letter was not strongly identified by parents as influencing their response. Conclusions: Personal contact with the person(s) providing information and requesting consent has a greater impact on parental consent responses than written information provided alone. This demonstrates that traditional bio-ethical ideas about rational decision-making where emotions are transcended and interference is not justified unless preventing harm to an unaware person are outdated. Parental decision-making is relational and the consent process should be adapted to reflect this. The current system that has a deontology view of decision making at its core impoverishes parental autonomy and may, ultimately, increase dental inequalities as a result.

Keywords: consent, decision, ethics, fluoride, parents

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Authors: Yaser S. Kishawi, Sadi R. Ali

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Part of Palestine, Gaza Strip (365 km2 and 1.8 million inhabitants) is considered a semi-arid zone relies solely on the Coastal Aquifer. The coastal aquifer is only source of water with only 5-10% suitable for human use. This barely cover the domestic and agricultural needs of Gaza Strip. Palestinian Water Authority Strategy is finding non-conventional water resource from treated wastewater to cover agricultural requirements and serve the population. A new WWTP project is to replace the old-overloaded Biet Lahia WWTP. The project consists of three parts; phase A (pressure line & infiltration basins - IBs), phase B (a new WWTP) and phase C (Recovery and Reuse Scheme – RRS – to capture the spreading plume). Currently, only phase A is functioning. Nearly 23 Mm3 of partially treated wastewater were infiltrated into the aquifer. Phase B and phase C witnessed many delays and this forced a reassessment of the RRS original design. An Environmental Management Plan was conducted from Jul 2013 to Jun 2014 on 13 existing monitoring wells surrounding the project location. This is to measure the efficiency of the SAT system and the spread of the contamination plume with relation to the efficiency of the proposed RRS. Along with the proposed location of the 27 recovery wells as part of the proposed RRS. The results of monitored wells were assessed compared with PWA baseline data. This was put into a groundwater model to simulate the plume to propose the best suitable solution to the delays. The redesign mainly manipulated the pumping rate of wells, proposed locations and functioning schedules (including wells groupings). The proposed simulations were examined using visual MODFLOW V4.2 to simulate the results. The results of monitored wells were assessed based on the location of the monitoring wells related to the proposed recovery wells locations (200m, 500m and 750m away from the IBs). Near the 500m line (the first row of proposed recovery wells), an increase of nitrate (from 30 to 70mg/L) compare to a decrease in Chloride (1500 to below 900mg/L) was found during the monitoring period which indicated an expansion of plume to this distance. On this rate with the required time to construct the recovery scheme, keeping the original design the RRS will fail to capture the plume. Based on that many simulations were conducted leading into three main scenarios. The scenarios manipulated the starting dates, the pumping rate and the locations of recovery wells. A simulation of plume expansion and path-lines were extracted from the model monitoring how to prevent the expansion towards the nearby municipal wells. It was concluded that the location is the most important factor in determining the RRS efficiency. Scenario III was adopted and showed an effective results even with a reduced pumping rates. This scenario proposed adding two additional recovery wells in a location beyond the 750m line to compensate the delays and effectively capture the plume. A continuous monitoring program for current and future monitoring wells should be in place to support the proposed scenario and ensure maximum protection.

Keywords: soil aquifer treatment, recovery and reuse scheme, infiltration basins, north gaza

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Authors: G. Kontochristopoulos, T. Spiliopoulos, V. Markantoni, E. Platsidaki, A. Kouris, E. Balamoti, C. Bokotas, G. Haidemenos

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Introduction: There is a high patient demand for periorbital rejuvenation since the facial area is often the first to show visible signs of aging. With advancing age, there are sometimes marked changes that occur in the skin, fat, muscle and bone of the periorbital region, resulting to wrinkles and skin laxity. These changes are among the easiest areas to correct using several minimally invasive techniques, which have become increasingly popular over the last decade. Lasers, radiofrequency, botulinum toxin, fat grafting and fillers are available treatments sometimes in combination to traditional blepharoplasty. This study attempts to show the benefits of a minimally invasive approach to periorbital wrinkles and skin laxity that combine microneedling and 10% trichloroacetic acid (TCA) peels. Method: Eleven female patients aged 34-72 enrolled in the study. They all gave informed consent after receiving detailed information regarding the treatment procedure. Exclusion criteria in the study were previous treatment for the same condition in the past six months, pregnancy, allergy or hypersensitivity to the components, infection, inflammation and photosensitivity on the affected region. All patients had diffuse periorbital wrinkles and mild to moderate upper or lower eyelid skin laxity. They were treated with Automatic Microneedle Therapy System-Handhold and topical application of 10% trichloroacetic acid solution to each periorbital area for five minutes. Needling at a 0,25 mm depth was performed in both latelar (x-y) directions. Subsequently, the peeling agent was applied to each periorbital area for five minutes. Patients were subjected to the above combination every two weeks for a series of four treatments. Subsequently they were followed up regularly every month for two months. The effect was photo-documented. A Physician's and a Patient's Global Assessment Scale was used to evaluate the efficacy of the treatment (0-25% indicated poor response, 25%-50% fair, 50%-75% good and 75%-100% excellent response). Safety was assessed by monitoring early and delayed adverse events. Results: At the end of the study, almost all patients demonstrated significant aesthetic improvement. Physicians assessed a fair and a good improvement in 9(81.8% of patients) and 2(18.1% of patients) participants respectively. Patients Global Assessment rated a fair and a good response in 6 (54.5%) and 5 (45.4%) participants respectively. The procedure was well tolerated and all patients were satisfied. Mild discomfort and transient erythema were quite common during or immediately after the procedure, however only temporary. During the monthly follow up, no complications or scars were observed. Conclusions: Microneedling is known as a simple, office–based collagen induction therapy. Low concentration TCA solution applied to the epidermis that has been more permeable by microneedling, can reach the dermis more effectively. In the present study, chemical peels with 10% TCA acted as an adjuvant to microneedling, as it causes controlled skin damage, promoting regeneration and rejuvenation of tissues. This combined therapy improved periorbital fine lines, wrinkles, and overall appearance of the skin. Thus it constitutes an alternative treatment of periorbital skin aging, with encouraging results and minor side-effects.

Keywords: chemical peels, microneedling, periorbital wrinkles, skin laxity

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Authors: Rana Mohamed, Ahmed E. Gomaa, Gehan Safwat, Ayman Diab

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Background: Bio-fabrication is a multidisciplinary research field that combines several principles, fabrication techniques, and protocols from different fields. The open-source-software movement is a movement that supports the use of open-source licenses for some or all software as part of the broader notion of open collaboration. Additive manufacturing is the concept of 3D printing, where it is a manufacturing method through adding layer-by-layer using computer-aided designs (CAD). There are several types of AM system used, and they can be categorized by the type of process used. One of these AM technologies is Digital light processing (DLP) which is a 3D printing technology used to rapidly cure a photopolymer resin to create hard scaffolds. DLP uses a projected light source to cure (Harden or crosslinking) the entire layer at once. Current applications of DLP are focused on dental and medical applications. Other developments have been made in this field, leading to the revolutionary field 3D bioprinting. The open-source movement was started to spread the concept of open-source software to provide software or hardware that is cheaper, reliable, and has better quality. Objective: Modification of desktop 3D printer into 3D bio-printer and the integration of DLP technology and bio-fabrication to produce an antibacterial dental model. Method: Modification of a desktop 3D printer into a 3D bioprinter. Gelatin hydrogel and sodium alginate hydrogel were prepared with different concentrations. Rhizome of Zingiber officinale, Flower buds of Syzygium aromaticum, and Bulbs of Allium sativum were extracted, and extractions were selected on different levels (Powder, aqueous extracts, total oils, and Essential oils) prepared for antibacterial bioactivity. Agar well diffusion method along with the E. coli have been used to perform the sensitivity test for the antibacterial activity of the extracts acquired by Zingiber officinale, Syzygium aromaticum, and Allium sativum. Lastly, DLP printing was performed to produce several dental models with the natural extracted combined with hydrogel to represent and simulate the Hard and Soft tissues. Result: The desktop 3D printer was modified into 3D bioprinter using open-source software Marline and modified custom-made 3D printed parts. Sodium alginate hydrogel and gelatin hydrogel were prepared at 5% (w/v), 10% (w/v), and 15%(w/v). Resin integration with the natural extracts of Rhizome of Zingiber officinale, Flower buds of Syzygium aromaticum, and Bulbs of Allium sativum was done following the percentage 1- 3% for each extract. Finally, the Antimicrobial dental model was printed; exhibits the antimicrobial activity, followed by merging with sodium alginate hydrogel. Conclusion: The open-source movement was successful in modifying and producing a low-cost Desktop 3D Bioprinter showing the potential of further enhancement in such scope. Additionally, the potential of integrating the DLP technology with bioprinting is a promising step toward the usage of the antimicrobial activity using natural products.

Keywords: 3D printing, 3D bio-printing, DLP, hydrogel, antibacterial activity, zingiber officinale, syzygium aromaticum, allium sativum, panax ginseng, dental applications

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Authors: Lauren B. Birney

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This project focuses upon the expansion of the existing "Curriculum and Community Enterprise for the Restoration of New York Harbor in New York City Public Schools" NSF EHR DRL 1440869, NSF EHR DRL 1839656 and NSF EHR DRL 1759006. This project is recognized locally as “Curriculum and Community Enterprise for Restoration Science,” or CCERS. CCERS is a comprehensive model of ecological restoration-based STEM education for urban public-school students. Following an accelerated rollout, CCERS is now being implemented in 120+ Title 1 funded NYC Department of Education middle schools, led by two cohorts of 250 teachers, serving more than 11,000 students in total. Initial results and baseline data suggest that the CCERS model, with the Billion Oyster Project (BOP) as its local restoration ecology-based STEM curriculum, is having profound impacts on students, teachers, school leaders, and the broader community of CCERS participants and stakeholders. Students and teachers report being receptive to the CCERS model and deeply engaged in the initial phase of curriculum development, citizen science data collection, and student-centered, problem-based STEM learning. The BOP CCERS Digital Platform will serve as the central technology hub for all research, data, data analysis, resources, materials and student data to promote global interactions between communities, Research conducted included qualitative and quantitative data analysis. We continue to work internally on making edits and changes to accommodate a dynamic society. The STEM Collaboratory NYC® at Pace University New York City continues to act as the prime institution for the BOP CCERS project since the project’s inception in 2014. The project continues to strive to provide opportunities in STEM for underrepresented and underserved populations in New York City. The replicable model serves as an opportunity for other entities to create this type of collaboration within their own communities and ignite a community to come together and address the notable issue. Providing opportunities for young students to engage in community initiatives allows for a more cohesive set of stakeholders, ability for young people to network and provide additional resources for those students in need of additional support, resources and structure. The project has planted more than 47 million oysters across 12 acres and 15 reef sites, with the help of more than 8,000 students and 10,000 volunteers. Additional enhancements and features on the BOP CCERS Digital Platform will continue over the next three years through funding provided by the National Science Foundation, NSF DRL EHR 1759006/1839656 Principal Investigator Dr. Lauren Birney, Professor Pace University. Early results from the data indicate that the new version of the Platform is creating traction both nationally and internationally among community stakeholders and constituents. This project continues to focus on new collaborative partners that will support underrepresented students in STEM Education. The advanced Digital Platform will allow for us connect with other countries and networks on a larger Global scale.

Keywords: STEM education, environmental restoration science, technology, citizen science

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Authors: Meiling Yu, Nadia Rouatbi, Khuloud T. Al-Jamal

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Background: Treatment of neurological disorders with modern medical and surgical approaches remains difficult. Gene therapy, allowing the delivery of genetic materials that encodes potential therapeutic molecules, represents an attractive option. The treatment of brain diseases with gene therapy requires the gene-editing tool to be delivered efficiently to the central nervous system. In this study, we explored the efficiency of different delivery routes, namely intravenous (i.v.), intra-cranial (i.c.), and intra-nasal (i.n.), to deliver stable nucleic acid-lipid particles (SNALPs) containing gene-editing tools namely Cas9 mRNA and sgRNA encoding for GFP as a reporter protein. We hypothesise that SNALPs can reach the brain and perform gene-editing to different extents depending on the administration route. Intranasal administration (i.n.) offers an attractive and non-invasive way to access the brain circumventing the blood–brain barrier. Successful delivery of gene-editing tools to the brain offers a great opportunity for therapeutic target validation and nucleic acids therapeutics delivery to improve treatment options for a range of neurodegenerative diseases. In this study, we utilised Rosa26-Cas9 knock-in mice, expressing GFP, to study brain distribution and gene-editing efficiency of SNALPs after i.v.; i.c. and i.n. routes of administration. Methods: Single guide RNA (sgRNA) against GFP has been designed and validated by in vitro nuclease assay. SNALPs were formulated and characterised using dynamic light scattering. The encapsulation efficiency of nucleic acids (NA) was measured by RiboGreen™ assay. SNALPs were incubated in serum to assess their ability to protect NA from degradation. Rosa26-Cas9 knock-in mice were i.v., i.n., or i.c. administered with SNALPs to test in vivo gene-editing (GFP knockout) efficiency. SNALPs were given as three doses of 0.64 mg/kg sgGFP following i.v. and i.n. or a single dose of 0.25 mg/kg sgGFP following i.c.. knockout efficiency was assessed after seven days using Sanger Sequencing and Inference of CRISPR Edits (ICE) analysis. In vivo, the biodistribution of DiR labelled SNALPs (SNALPs-DiR) was assessed at 24h post-administration using IVIS Lumina Series III. Results: Serum-stable SNALPs produced were 130-140 nm in diameter with ~90% nucleic acid loading efficiency. SNALPs could reach and stay in the brain for up to 24h following i.v.; i.n. and i.c. administration. Decreasing GFP expression (around 50% after i.v. and i.c. and 20% following i.n.) was confirmed by optical imaging. Despite the small number of mice used, ICE analysis confirmed GFP knockout in mice brains. Additional studies are currently taking place to increase mice numbers. Conclusion: Results confirmed efficient gene knockout achieved by SNALPs in Rosa26-Cas9 knock-in mice expressing GFP following different routes of administrations in the following order i.v.= i.c.> i.n. Each of the administration routes has its pros and cons. The next stages of the project involve assessing gene-editing efficiency in wild-type mice and replacing GFP as a model target with therapeutic target genes implicated in Motor Neuron Disease pathology.

Keywords: CRISPR, nanoparticles, brain diseases, administration routes

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Authors: Rafia S. Al-lamki, Jun Wang, Simon Pacey, Jordan Pober, John R. Bradley

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Tumor necrosis factor alpha (TNF), signaling through TNFR2, may act an autocrine growth factor for renal tubular epithelial cells. Clear cell renal carcinomas (ccRCC) contain cancer stem cells (CSCs) that give rise to progeny which form the bulk of the tumor. CSCs are rarely in cell cycle and, as non-proliferating cells, resist most chemotherapeutic agents. Thus, recurrence after chemotherapy may result from the survival of CSCs. Therapeutic targeting of both CSCs and the more differentiated bulk tumor populations may provide a more effective strategy for treatment of RCC. In this study, we hypothesized that TNFR2 signaling will induce CSCs in ccRCC to enter cell cycle so that treatment with ligands that engage TNFR2 will render CSCs susceptible to chemotherapy. To test this hypothesis, we have utilized wild-type TNF (wtTNF) or specific muteins selective for TNFR1 (R1TNF) or TNFR2 (R2TNF) to treat either short-term organ cultures of ccRCC and adjacent normal kidney (NK) tissue or cultures of CD133+ cells isolated from ccRCC and adjacent NK, hereafter referred to as stem cell-like cells (SCLCs). The effect of cyclophosphamide (CP), currently an effective anticancer agent, was tested on CD133+SCLCs from ccRCC and NK before and after R2TNF treatment. Responses to TNF were assessed by flow cytometry (FACS), immunofluorescence, and quantitative real-time PCR, TUNEL, and cell viability assays. Cytotoxic effect of CP was analyzed by Annexin V and propidium iodide staining with FACS. In addition, we assessed the effect of TNF on isolated SCLCs differentiation using a three-dimensional (3D) culture system. Clinical samples of ccRCC contain a greater number SCLCs compared to NK and the number of SCSC increases with higher tumor grade. Isolated SCLCs show expression of stemness markers (oct4, Nanog, Sox2, Lin28) but not differentiation markers (cytokeratin, CD31, CD45, and EpCAM). In ccRCC organ cultures, wtTNF and R2TNF increase CD133 and TNFR2 expression and promote cell cycle entry whereas wtTNF and R1TNF increase TNFR1 expression and promote cell death of SCLCs. Similar findings are observed in SCLCs isolated from NK but the effect was greater in SCLCs isolated from ccRCC. Application of CP distinctly triggered apoptotic and necrotic cell death in SLCSs pre-treatment with R2TNF as compared to CP treatment alone, with SCLCs from ccRCC more sensitive to CP compared to SLCS from NK. Furthermore, TNF promotes differentiation of SCLCs to an epithelial phenotype in 3D cultures, confirmed by cytokeratin expression and loss of stemness markers Nanog and Sox2. The differentiated cells show positive expression of TNF and TNFR2. These findings provide evidence that selective engagement of TNFR2 drive CSCs to cell proliferation/differentiation, and targeting of cycling cells with TNFR2 agonist in combination with anti-cancer agents may be a potential therapy for RCC.

Keywords: cancer stem cells, ccRCC, cell cycle, cell death, TNF, TNFR1, TNFR2, CD133

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