Search results for: temporal resolution

Authors: Shane Theunissen, Rob Grandy

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Historically and we would fairly conventionally within our formal schooling systems, we have convergent testing where all the students are expected to converge on the same answer, and that answer has been determined by an external authority that is reproducing knowledge of the hegemon. Many youths may not embody the cultural capital that's rewarded in formal schooling contexts as they aren't able to converge on the required answer that's being determined by the classroom teacher or the administrators. In this paper, we explore divergent processes that promote creative problem-solving. We embody this divergent process in our measurement of fundamental growth needs. To this end, we utilize the Mosaic Approach as a method for implementing the Outcomes That Matter framework. Outcomes That Matter is the name of the measurement tool built around the Circle of Courage framework, which is a way of identifying fundamental growth needs for young people. The Circle of Courage was developed by Martin-Broken-Leg and colleagues as a way to connect indigenous child-rearing philosophies with contemporary resilience and positive psychology research. The Outcomes that Matter framework puts forward four categories of growth needs for young people. These are: Belonging, which on a macro scale is acceptance into the greater community of practice, Mastery which includes a constellation of concepts including confidence, motivation, self-actualization, and self-determination, Independence refers to a sense of personal power into autonomy within a context where creativity and problem solving, and a personal voice can begin to emerge, and finally Generosity which includes interpersonal things like conflict resolution and teamwork. Outcomes of Matter puts these four domains into a measurement tool that facilitates collaborative assessment between the youth, teachers, and recreation therapists that allows for youth-led narratives pertaining to their fundamental growth outcomes. This application of the Outcomes That Matter framework is unique as it may be the first application of this framework in an educational boatbuilding context.

Keywords: collaboration, empowerment, outcomes that matter, mosaic approach, boat building

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Authors: Gul Jan

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It is the first quantitative ethno-botanical analysis and conservation issues of medicinal flora of Alpine and Sub-alpine, Hindikush region of Pakistan. The objective of the study aims to report, compare the uses and highlight the ethno-Botanical significance of medicinal plants for treatment of various diseases. A total of 250 (242 males and 8 females) local informants including 10 Local Traditional Healers were interviewed. Information was collected through semi-structured interviews, analyzed and compared by quantitative ethno-botanical indices such as Jaccard index (JI), Informant Consensus Factor (ICF), use value (UV) and Relative frequency of citation (RFC).Thorough survey indicated that 57 medicinal plants belongs to 43 families were investigated to treat various illnesses. The highest ICF is recorded for digestive system (0.69%), Circolatory system (0.61%), urinary tract system, (0.53%) and respiratory system (0.52%). Used value indicated that, Achillea mellefolium (UV = 0.68), Aconitum violaceum (UV = 0.69), Valeriana jatamansi (UV = 0.63), Berberis lyceum (UV = 0.65) and are exceedingly medicinal plant species used in the region. In comparison, highest similarity index is recorded in these studies with JI 17.72 followed by 16.41. According to DMR output, Pinus williciana ranked first due to multipurpose uses among all species and was found most threatened with higher market value. Unwise used of natural assets pooled with unsuitable harvesting practices have exaggerated pressure on plant species of the research region. The main issues causative to natural variety loss found were over grazing of animals, forest violation, wild animal hunting, fodder, plant collection as medicine, fuel wood, forest fire, and invasive species negatively affect the natural resources. For viable utilization, in situ and ex situ conservation, skillful collecting, and reforestation project may be the resolution. Further wide field management research is required.

Keywords: quantitative analysis, conservations issues, medicinal flora, alpine and sub-alpine, Hindukush region

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Authors: Chiao-Yi Chen, Dung-Ying Lin

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With the growing awareness of environmental protection and the implementation of government carbon reduction policies, the number of electric vehicles (EVs) has rapidly increased, leading to a surge in charging demand and imposing significant challenges on the existing power grid’s capacity. Traditional urban power grid planning has not adequately accounted for the additional load generated by EV charging, which often strains the infrastructure. This study aims to optimize grid operation and load management by dynamically adjusting EV charging prices based on real-time electricity supply and demand, leveraging consumer demand elasticity to enhance system efficiency. This study uniquely addresses the intricate interplay between urban traffic patterns and power grid dynamics in the context of electric vehicle (EV) adoption. By integrating Hsinchu City's road network with the IEEE 33-bus system, the research creates a comprehensive model that captures both the spatial and temporal aspects of EV charging demand. This approach allows for a nuanced analysis of how traffic flow directly influences the load distribution across the power grid. The strategic placement of charging stations at key nodes within the IEEE 33-bus system, informed by actual road traffic data, enables a realistic simulation of the dynamic relationship between vehicle movement and energy consumption. This integration of transportation and energy systems provides a holistic view of the challenges and opportunities in urban EV infrastructure planning, highlighting the critical need for solutions that can adapt to the ever-changing interplay between traffic patterns and grid capacity. The proposed dynamic pricing strategy effectively reduces peak charging loads, enhances the operational efficiency of charging stations, and maximizes operator profits, all while ensuring grid stability. These findings provide practical insights and a valuable framework for optimizing EV charging infrastructure and policies in future smart cities, contributing to more resilient and sustainable urban energy systems.

Keywords: dynamic pricing, demand elasticity, EV charging, grid load balancing, optimization

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Authors: Adi Gielgun-Katz, Alina S. Rusu

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In the last two decades, education systems worldwide have integrated new pedagogical methods and strategies in lesson plans, such as innovative technologies, social-emotional learning (SEL), gamification, mixed learning, multiple literacies, and many others. Visual language, such as photographs, is known to transcend cultures and languages, and it is commonly used by youth to express positions and affective states in social networks. Therefore, visual language needs more educational attention as a linguistic and communicative component that can create connectedness among the students and their teachers. Nowadays, when SEL is gaining more and more space and meaning in the area of academic improvement in relation to social well-being, and taking and sharing pictures is part of the everyday life of the majority of people, it becomes natural to add the visual language to SEL approach as a reinforcement strategy for connecting education to the contemporary culture and language of the youth. This article presents a program conducted in a high school class in Israel, which combines the five SEL with photography techniques, i.e., Social-Emotional Learning Visual Empowerments (SELVEs) program (experimental group). Another class of students from the same institution represents the control group, which is participating in the SEL program without the photography component. The SEL component of the programs addresses skills such as: troubleshooting, uncertainty, personal strengths and collaboration, accepting others, control of impulses, communication, self-perception, and conflict resolution. The aim of the study is to examine the effects of programs on the level of the five SEL aspects in the two groups of high school students: Self-Awareness, Social Awareness, Self-Management, Responsible Decision Making, and Relationship Skills. The study presents a quantitative assessment of the SEL programs’ impact on the students. The main hypothesis is that the students’ questionnaires' analysis will reveal a better understanding and improvement of the five aspects of the SEL in the group of students involved in the photography-enhanced SEL program.

Keywords: social-emotional learning, photography, education program, adolescents

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Authors: Alexander N. Savostyanov, Tatyana A. Dolgorukova, Elena A. Esipenko, Mikhail S. Zaleshin, Margherita Malanchini, Anna V. Budakova, Alexander E. Saprygin, Yulia V. Kovas

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The present study compared spectral-power indexes and cortical topography of brain activity in a sample characterized by different levels of trait and mathematical anxiety. 52 healthy Russian-speakers (age 17-32; 30 males) participated in the study. Participants solved an error recognition task under 3 conditions: A lexical condition (simple sentences in Russian), and two numerical conditions (simple arithmetic and complicated algebraic problems). Trait and mathematical anxiety were measured using self-repot questionnaires. EEG activity was recorded simultaneously during task execution. Event-related spectral perturbations (ERSP) were used to analyze spectral-power changes in brain activity. Additionally, sLORETA was applied in order to localize the sources of brain activity. When exploring EEG activity recorded after tasks onset during lexical conditions, sLORETA revealed increased activation in frontal and left temporal cortical areas, mainly in the alpha/beta frequency ranges. When examining the EEG activity recorded after task onset during arithmetic and algebraic conditions, additional activation in delta/theta band in the right parietal cortex was observed. The ERSP plots reveled alpha/beta desynchronizations within a 500-3000 ms interval after task onset and slow-wave synchronization within an interval of 150-350 ms. Amplitudes of these intervals reflected the accuracy of error recognition, and were differently associated with the three (lexical, arithmetic and algebraic) conditions. The level of trait anxiety was positively correlated with the amplitude of alpha/beta desynchronization. The level of mathematical anxiety was negatively correlated with the amplitude of theta synchronization and of alpha/beta desynchronization. Overall, trait anxiety was related with an increase in brain activation during task execution, whereas mathematical anxiety was associated with increased inhibitory-related activity. We gratefully acknowledge the support from the №11.G34.31.0043 grant from the Government of the Russian Federation.

Keywords: anxiety, EEG, lexical and numerical error-recognition tasks, alpha/beta desynchronization

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Authors: Ralene Sim, Stefan Mueller, N. Gopalakrishna Iyer, Ngian Chye Tan, Khee Chee Soo, R. Shetty Mahalakshmi, Hiang Khoon Tan

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Background: Regional failure in nasopharyngeal carcinoma (NPC) is managed by salvage treatment in the form of neck dissection. Radical neck dissection (RND) is preferred over modified radical neck dissection (MRND) since it is traditionally believed to offer better long-term disease control. However, with the advent of more advanced imaging modalities like high-resolution Magnetic Resonance Imaging, Computed Tomography, and Positron Emission Tomography-CT scans, earlier detection is achieved. Additionally, concurrent chemotherapy also contributes to reduced tumour burden. Hence, there may be a lesser need for an RND and a greater role for MRND. With this retrospective study, the primary aim is to ascertain whether MRND, as opposed to RND, has similar outcomes and hence, whether there would be more grounds to offer a less aggressive procedure to achieve lower patient morbidity. Methods: This is a retrospective study of 66 NPC patients treated at Singapore General Hospital between 1994 to 2016 for histologically proven regional recurrence, of which 41 patients underwent RND and 25 who underwent MRND, based on surgeon preference. The type of ND performed, primary treatment mode, adjuvant treatment, and pattern of recurrence were reviewed. Overall survival (OS) was calculated using Kaplan-Meier estimate and compared. Results: Overall, the disease parameters such as nodal involvement and extranodal extension were comparable between the two groups. Comparing MRND and RND, the median (IQR) OS is 1.76 (0.58 to 3.49) and 2.41 (0.78 to 4.11) respectively. However, the p-value found is 0.5301 and hence not statistically significant. Conclusion: RND is more aggressive and has been associated with greater morbidity. Hence, with similar outcomes, MRND could be an alternative salvage procedure for regional failure in selected NPC patients, allowing similar salvage rates with lesser mortality and morbidity.

Keywords: nasopharyngeal carcinoma, neck dissection, modified neck dissection, radical neck dissection

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Authors: Abduladim Maitieg, Mark Johnson

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Developing a coastal oil spill management plan for the Misratah coast is the motivating factor for building a database for coastal and marine systems and energy resources. Wind direction and speed, currents, bathymetry, coastal topography and offshore dynamics influence oil spill deposition in coastal water. Therefore, oceanographic and climatological data can be used to understand oil slick movement and potential oil deposits on shoreline area and the behaviour of oil spill trajectories on the sea surface. The purpose of this study is to investigate the effects of the coastal and marine physical factors under strong wave conditions and various bathymetric and coastal topography gradients in the western coastal area of Libya on the movement of oil slicks. The movement of oil slicks was computed using a GNOME simulation model based on current and wind speed/direction. The results in this paper show that (1) Oil slick might reach the Misratah shoreline area in two days in the summer and winter. Seasons. (2 ) The North coast of Misratah is the potential oil deposit area on the Misratah coast. (3) Tarball pollution was observed along the North coast of Misratah. (4) Two scenarios for the summer and the winter season were run, along the western coast of Libya . (5) The eastern coast is at a lower potential risk due to the influence of wind and current energy in the Gulf of Sidra. (6) The Misratah coastline is more vulnerable to oil spill movement in the summer than in winter seasons. (7) Oil slick takes from 2 to 5 days to reach the saltmarsh in the eastern Misratah coast. (8) Oil slick moves 300 km in 30 days from the spill resource location near the Libyan western border to the Misratah coast.(9) Bathymetric features have a profound effect on oil spill movement. (9)Oil dispersion simulations using GNOME are carried out taking into account high-resolution wind and current data.

Keywords: oil spill movement, coastal and marine physical factors, coast area, Libyan

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Authors: Vistorina Amputu, Katja Tielboerger, Nichola Knox

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Primary productivity in dry savannahs is constraint by moisture availability and under increasing anthropogenic pressure. Thus, considering climate change and the unprecedented pace and scale of rangeland deterioration, methods for assessing the status of such rangelands should be easy to apply, yield reliable and repeatable results that can be applied over large spatial scales. Global and local scale monitoring of rangelands through satellite data and labor-intensive field measurements respectively, are limited in accurately assessing the spatiotemporal heterogeneity of vegetation dynamics to provide crucial information that detects degradation in its early stages. Fortunately, newly emerging techniques such as unmanned aerial vehicles (UAVs), associated miniaturized sensors and improving digital photogrammetric software provide an opportunity to transcend these limitations. Yet, they have not been extensively calibrated in natural systems to encompass their complexities if they are to be integrated for long-term monitoring. Limited research using drone technology has been conducted in arid savannas, for example to assess the health status of this dynamic two-layer vegetation ecosystem. In our study, we fill this gap by testing the relationship between UAV-estimated cover of rangeland functional attributes and field data collected in discrete sample plots in a Namibian dryland savannah along a degradation gradient. The first results are based on a supervised classification performed on the ultra-high resolution multispectral imagery to distinguish between rangeland functional attributes (bare, non-woody, and woody), with a relatively good match to the field observations. Integrating UAV-based observations to improve rangeland monitoring could greatly assist in climate-adapted rangeland management.

Keywords: arid savannah, degradation gradient, field observations, narrow-band sensor, supervised classification

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Authors: Sidrah Ahmed

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The river and lakes flows are modeled mathematically by shallow water equations that are depth-averaged Reynolds Averaged Navier-Stokes equations under Boussinesq approximation. The temperature stratification dynamics influence the water quality and mixing characteristics. It is mainly due to the atmospheric conditions including air temperature, wind velocity, and radiative forcing. The experimental observations are commonly taken along vertical scales and are not sufficient to estimate small turbulence effects of temperature variations induced characteristics of shallow flows. Wind shear stress over the water surface influence flow patterns, heat fluxes and thermodynamics of water bodies as well. Hence it is crucial to couple temperature gradients with shallow water model to estimate the atmospheric effects on flow patterns. The Ripa system has been introduced to study ocean currents as a variant of shallow water equations with addition of temperature variations within the flow. Ripa model is a hyperbolic system of partial differential equations because all the eigenvalues of the system’s Jacobian matrix are real and distinct. The time steps of a numerical scheme are estimated with the eigenvalues of the system. The solution to Riemann problem of the Ripa model is composed of shocks, contact and rarefaction waves. Solving Ripa model with Riemann initial data with the central schemes is difficult due to the eigen structure of the system.This works presents the comparison of four different finite difference schemes for the numerical solution of Riemann problem for Ripa model. These schemes include Lax-Friedrichs, Lax-Wendroff, MacCormack scheme and a higher order finite difference scheme with WENO method. The numerical flux functions in both dimensions are approximated according to these methods. The temporal accuracy is achieved by employing TVD Runge Kutta method. The numerical tests are presented to examine the accuracy and robustness of the applied methods. It is revealed that Lax-Freidrichs scheme produces results with oscillations while Lax-Wendroff and higher order difference scheme produce quite better results.

Keywords: finite difference schemes, Riemann problem, shallow water equations, temperature gradients

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Authors: Joel F. Campbell, Bing Lin, Michael Obland, Susan Kooi, Tai-Fang Fan, Byron Meadows, Edward Browell, Wayne Erxleben, Doug McGregor, Jeremy Dobler, Sandip Pal, Christopher O'Dell, Ken Davis

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The Active Sensing of CO₂ Emissions over Nights, Days, and Seasons (ASCENDS) CarbonHawk Experiment Simulator (ACES) is a NASA Langley Research Center instrument funded by NASA’s Science Mission Directorate that seeks to advance technologies critical to measuring atmospheric column carbon dioxide (CO₂ ) mixing ratios in support of the NASA ASCENDS mission. The ACES instrument, an Intensity-Modulated Continuous-Wave (IM-CW) lidar, was designed for high-altitude aircraft operations and can be directly applied to space instrumentation to meet the ASCENDS mission requirements. The ACES design demonstrates advanced technologies critical for developing an airborne simulator and spaceborne instrument with lower platform consumption of size, mass, and power, and with improved performance. The Atmospheric Carbon and Transport – America (ACT-America) is an Earth Venture Suborbital -2 (EVS-2) mission sponsored by the Earth Science Division of NASA’s Science Mission Directorate. A major objective is to enhance knowledge of the sources/sinks and transport of atmospheric CO₂ through the application of remote and in situ airborne measurements of CO₂ and other atmospheric properties on spatial and temporal scales. ACT-America consists of five campaigns to measure regional carbon and evaluate transport under various meteorological conditions in three regional areas of the Continental United States. Regional CO₂ distributions of the lower atmosphere were observed from the C-130 aircraft by the Harris Corp. Multi-Frequency Fiber Laser Lidar (MFLL) and the ACES lidar. The airborne lidars provide unique data that complement the more traditional in situ sensors. This presentation shows the applications of CO₂ lidars in support of these science needs.

Keywords: CO₂ measurement, IMCW, CW lidar, laser spectroscopy

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Authors: Mohammed M. Mahdy Tawfeek

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Objective: The aim of this work is to compare the outcome of photoactivated chromophore for keratitis-cross linking (PACK-CXL) window absorption (WA) alone with combined PACK-CXL WA and standard anti-microbial therapy (SAT) for the treatment of infectious keratitis. Patients and Methods: This is a randomized prospective comparative clinical trial. Thirty eyes with clinically suspected infectious keratitis were randomly assigned into two equal groups of 15 eyes each: Group (A) was treated by PACK-CXL WA alone and group (B) was treated by PACK-CXL WA combined with SAT. Identification of organisms was made by lab study before treatment. Corneal healing was evaluated by corneal examination and anterior segment OCT (AS-OCT). Written informed consent was obtained from all participants and the study was approved by the research ethics committee of the Faculty of Medicine, Zagazig University. The work has been carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki) for studies involving humans. Results: Complete healing and resolution (Successful treatment) were observed in 10 eyes (66.7%) of a group (A) and 14 eyes (93.3%) of group (B) and failure was observed in 5 eyes (33.3%) of a group (A) and one eye (6.67%) of group (B). They were statistically significant (P =0.042 and 0.003) in a comparison between both groups regarding success and failure of treatment, respectively. Complete corneal healing was reported in the third month postoperatively in 10 eyes (66.7%) of group (A) and 14 eyes (93.3%) of group (B). Complications were absent in 12 patients (80%) in group (A) and 14 patients (93.3%) of group (B); however, perforation and impending perforation were found in 3 patients of group (A) and only one patient of group (B). Conclusion: PACK-CXL is a promising, non-invasive treatment option for infectious keratitis, especially when performed with the window absorption (WA) technique, either alone or combined with SAT. It has a synergistic effect with a standard antimicrobial treatment that gives good outcome results in the treatment of infectious keratitis. Also, it avoids the antibiotics resistance that has become rapidly spreading worldwide.

Keywords: corneal cross linking, infectious keratitis, PACK-CXL, window absorption

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Authors: Ekaterina Sánchez Romero, Emily Gabriela Aguirre Herrera, Sandra Luz Espinoza Esquerra, Jorge García Campos

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Female, 7 months old, daughter of a mother with anemia during pregnancy, with no history of atopy in the family, since birth she presents with recurrent dermatological and gastrointestinal infections, chronically treated for recurrent diaper dermatitis. At 6 months of age, she begins with generalized pallor, hyperpigmentation in hands and feet, smooth tongue, psychomotor retardation with lack of head support, sedation, and hypoactivity. She was referred to our hospital for a fever of 38°C, severe diaper rash, and pancytopenia with HB 9.3, platelets 38000, neutrophils 0.39 MCV: 86.80 high for her age. The approach was initiated to rule out myeloproliferative syndrome, with negative immunohistochemical results of bone marrow aspirate; during her stay, she presented neurological regression, lack of sucking, and focal seizures. CT scan showed cortical atrophy. The patient was diagnosed with primary immunodeficiency due to history; gamma globulin was administered without improvement with normal results of immunoglobulins and metabolic screening. When dermatological and neurological diagnoses were ruled out as the primary cause, a nutritional factor was evaluated, and a therapeutic trial was started with the administration of vitamin B12 and zinc, presenting clinical neurological improvement and resolution of pancytopenia in 2 months. It was decided to continue outpatient management. Discussion: We present a patient with neurological, dermatological involvement, and pancytopenia, so the most common differential diagnoses in this population were ruled out. Vitamin B12 deficiency is an uncommon entity. Due to maternal and clinical history, a therapeutic trial was started resulting in an improvement. Conclusion: VitaminB12 deficiency should be considered one of the differential diagnoses in the approach to pancytopenia with megaloblastic anemia associated with dermatologic and neurologic manifestations. Early treatment can reduce irreversible damage in these patients.

Keywords: vitamin B12 deficiency, pediatrics, pancytopenia, diaper dermatitis

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Authors: Anand Malik

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The Himalayas with high tectonic activities poses a great threat to human life and property. Climate change is another reason which triggering extreme events multiple fold effect on high mountain glacial environment, rock falls, landslides, debris flows, flash flood and snow avalanches. One such extreme event of cloud burst along with breach of moraine dammed Chorabri Lake occurred from June 14 to June 17, 2013, triggered flooding of Saraswati and Mandakini rivers in the Kedarnath Valley of Rudraprayag district of Uttrakhand state of India. As a result, huge volume of water with its high velocity created a catastrophe of the century, which resulted into loss of large number of human/animals, pilgrimage, tourism, agriculture and property. Thus a comprehensive assessment of debris flow hazards requires GIS-based modeling using numerical methods. The aim of present study is to focus on analysis and mapping of debris flow movements using geospatial data with flow-r (developed by team at IGAR, University of Lausanne). The model is based on combined probabilistic and energetic algorithms for the assessment of spreading of flow with maximum run out distances. Aster Digital Elevation Model (DEM) with 30m x 30m cell size (resolution) is used as main geospatial data for preparing the run out assessment, while Landsat data is used to analyze land use land cover change in the study area. The results of the study area show that model can be applied with great accuracy as the model is very useful in determining debris flow areas. The results are compared with existing available landslides/debris flow maps. ArcGIS software is used in preparing run out susceptibility maps which can be used in debris flow mitigation and future land use planning.

Keywords: debris flow, geospatial data, GIS based modeling, flow-R

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Authors: Haiming Wen, Isabella J. Van Rooyen

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Tristructural isotropic (TRISO)-coated fuel particles are designed as nuclear fuel for high-temperature gas reactors. TRISO coating consists of layers of carbon buffer, inner pyrolytic carbon (IPyC), SiC, and outer pyrolytic carbon. The TRISO coating, especially the SiC layer, acts as a containment system for fission products produced in the kernel. However, release of certain metallic fission products across intact TRISO coatings has been observed for decades. Despite numerous studies, mechanisms by which fission products migrate across the coating layers remain poorly understood. In this study, scanning transmission electron microscopy (STEM), energy dispersive X-ray spectroscopy (EDS), high-resolution transmission electron microscopy (HRTEM) and electron energy loss spectroscopy (EELS) were used to examine the distribution, composition and structure of fission products in a TRISO coated particle neutron irradiated to 3.6 x 1021 n/cm² fast fluence at 1040 ⁰C. Precession electron diffraction was used to investigate characters of grain boundaries where specific fission product precipitates are located. The retention fraction of 110mAg in the investigated TRISO particle was estimated to be 0.19. A high density of nanoscale fission product precipitates was observed in the SiC layer close to the SiC-IPyC interface, most of which are rich in Pd, while Ag was not identified. Some Pd-rich precipitates contain U. Precipitates tend to have complex structure and composition. Although a precipitate appears to have uniform contrast in STEM, EDS indicated that there may be composition variations throughout the precipitate, and HRTEM suggested that the precipitate may have several parts different in crystal structure or orientation. Attempts were made to measure charge states of precipitates using EELS and study their possible effect on precipitate transport.

Keywords: TRISO particle, fission product, nuclear fuel, electron microscopy, neutron irradiation

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Authors: Maryam S. Ghoraishi, John E. Hawk, Thomas Thundat

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Understanding liquid properties in small scale has become important in recent decades as immerging new microelectromechanical systems (MEMS) devices have been widely used for micro pumps, drug delivery, and many other laboratory-on-microchips analysis. Often in microfluidic devices, fluids are transported electrokinetically. Therefore, extensive knowledge of fluid flow, heat transport, electrokinetics and electrochemistry are key to successful lab on a chip design. Among different microfluidic devices, recently developed hollow channel cantilever offers an ideal platform to study different fluid properties simultaneously without drastic decrease in quality factor which normally occurs when traditional cantilevers operate in the liquid phase. Using hollow channel cantilever, we monitor changes in density and viscosity of liquid while simultaneously investigating dielectric properties of alcohol water binary mixtures. Considerable research has been conducted on alcohol-water mixtures since such a mixture is a typical prototype for biomolecules, Micelle formation, and structural stability of proteins (to name a few). Here we show that hollow channel cantilever can be employed to investigate dielectric properties of ethanol/water mixtures in different concentrations. We study dynamic amplitude shifts of hollow channel cantilever oscillation at different concentrations of ethanol/water for different voltages. Our results show how interactions between solute and solvent, and possibly cluster formation, could change dielectric properties and dipole reorientation of the mixture, as well as the resulting force on the hollow cantilever. For comparison, we also examine higher conductivity ionic mixtures of sodium sulfate solution under the same conditions as low conductivity ethanol/water mixtures. We will show the results from systematic investigation of solvent effects on dielectric properties of the binary mixture. We will also address the question of resolution limits in dielectric study of analyte molecules imposed by solvent concentrations.

Keywords: dielectric constant, cantilever sensors, ethanol water mixtures, low frequency

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Authors: Benjamin Newman

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The picture of an adverse contest between two parties has often been used as an archetypal description of the Anglo-American adversarial criminal trial. However, in actuality, guilty pleas and plea-bargains have been dominating the procedure for over the last half-a-century. Characterised by two adverse parties, the court adjudicative system in the Anglo-American world adhere to the adversarial procedure, and while further features have been attributed and the values that are embedded within the procedure vary, it is a system that we have no adequate theory. Damaska had argued that the adversarial conflict-resolution mode of administration of justice stems from a liberal laissez-faire concept of a value neutral liberal state. Having said that, the court’s neutrality has been additionally rationalised in light of its liberal end as a safeguard from the state’s coercive force. Both conceptions of the court’s neutrality conflict in cases where the by-standing role disposes of its liberal duty in safeguarding the individual. Such is noticeable in plea bargains, where the defendant has the liberty to plead guilty, despite concerns over wrongful convictions and deprivation of liberty. It is an inner liberal tension within the notion of criminal adversarialism, between the laissez-faire mode which grants autonomy to the parties and the safeguarding liberal end of the trial. Langbein had asserted that the adversarial system is a criminal procedure for which we have no adequate theory, and it is by reference to political and moral theories that the research aims to articulate a normative account. The paper contemplates on the above liberal-tension, and by reference to Duff’s ‘calling-to-account’ theory, argues that autonomy is of inherent value to the criminal process, being considered a constitutive element in the process of being called to account. While the aspiration is that the defendant’s guilty plea should be genuine, the guilty-plea decision must be voluntary if it is to be considered a performative act of accountability. Thus, by valuing procedural autonomy as a necessary element within the criminal adjudicative process, it assimilates a liberal procedure, whilst maintaining the liberal end by holding the defendant to account.

Keywords: liberal theory, adversarial criminal procedure, criminal law theory, liberal perfectionism, political liberalism

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Authors: Emilio A. Berny-Brandt, Sonia E. Ruiz

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Steel tubular towers serving as support structures for large wind turbines are subject to several hundred million stress cycles arising from the turbulent nature of the wind. This causes high-cycle fatigue which can govern tower design. The practice of maintaining the support structure after wind turbines reach its typical 20-year design life have become common, but without quantifying the changes in the reliability on the tower. There are several studies on this topic, but most of them are based on the S-N curve approach using the Miner’s rule damage summation method, the de-facto standard in the wind industry. However, the qualitative nature of Miner’s method makes desirable the use of fracture mechanics to measure the effects of fatigue in the capacity curve of the structure, which is important in order to evaluate the integrity and reliability of these towers. Temporal and spatially varying wind speed time histories are simulated based on power spectral density and coherence functions. Simulations are then applied to a SAP2000 finite element model and step-by-step analysis is used to obtain the stress time histories for a range of representative wind speeds expected during service conditions of the wind turbine. Rainflow method is then used to obtain cycle and stress range information of each of these time histories and a statistical analysis is performed to obtain the distribution parameters of each variable. Monte Carlo simulation is used here to evaluate crack growth over time in the tower base using the Paris-Erdogan equation. A nonlinear static pushover analysis to assess the capacity curve of the structure after a number of years is performed. The capacity curves are then used to evaluate the changes in reliability of a steel tower located in Oaxaca, Mexico, where wind energy facilities are expected to grow in the near future. Results show that fatigue on the tower base can have significant effects on the structural capacity of the wind turbine, especially after the 20-year design life when the crack growth curve starts behaving exponentially.

Keywords: crack growth, fatigue, Monte Carlo simulation, structural reliability, wind turbines

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Authors: Tuba Zahra, Aakash Parekh

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Hyperspectral remote sensing refers to imaging of objects or materials in narrow conspicuous spectral bands. Hyperspectral images (HSI) enable the extraction of spectral signatures for objects or materials observed. These images contain information about the reflectance of each pixel across the electromagnetic spectrum. It enables the acquisition of data simultaneously in hundreds of spectral bands with narrow bandwidths and can provide detailed contiguous spectral curves that traditional multispectral sensors cannot offer. The contiguous, narrow bandwidth of hyperspectral data facilitates the detailed surveying of Earth's surface features. This would otherwise not be possible with the relatively coarse bandwidths acquired by other types of imaging sensors. Hyperspectral imaging provides significantly higher spectral and spatial resolution. There are several use cases that represent the commercial applications of hyperspectral remote sensing. Each use case represents just one of the ways that hyperspectral satellite imagery can support operational efficiency in the respective vertical. There are some use cases that are specific to VNIR bands, while others are specific to SWIR bands. This paper discusses the different commercially viable use cases that are significant for HSI application areas, such as agriculture, mining, oil and gas, defense, environment, and climate, to name a few. Theoretically, there is n number of use cases for each of the application areas, but an attempt has been made to streamline the use cases depending upon economic feasibility and commercial viability and present a review of literature from this perspective. Some of the specific use cases with respect to agriculture are crop species (sub variety) detection, soil health mapping, pre-symptomatic crop disease detection, invasive species detection, crop condition optimization, yield estimation, and supply chain monitoring at scale. Similarly, each of the industry verticals has a specific commercially viable use case that is discussed in the paper in detail.

Keywords: agriculture, mining, oil and gas, defense, environment and climate, hyperspectral, VNIR, SWIR

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Authors: Li-Wei Wu, Chung-Yu Wang

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Recently, interest has been generated in the search for a new reference framework for value creation that is centered on the co-creation process. Co-creation implies cooperative value creation between service firms and customers and requires the building of experiences as well as the resolution of problems through the combined effort of the parties in the relationship. For customers, values are always co-created through their participation in services. Customers can ultimately determine the value of the service in use. This new approach emphasizes that a customer’s participation in the service process is considered indispensable to value co-creation. An important feature of service in the context of exchange is co-production, which implies that a certain amount of participation is needed from customers to co-produce a service and hence co-create value. Co-production no doubt helps customers better understand and take charge of their own roles in the service process. Thus, this proposal is to encourage co-production, thus facilitating value co-creation of that is reflected in both customers and service firms. Four determinants of co-production are identified in this study, namely, commitment, trust, asset specificity, and decision-making uncertainty. Commitment is an essential dimension that directly results in successful cooperative behaviors. Trust helps establish a relational environment that is fundamental to cross-border cooperation. Asset specificity motivates co-production because this determinant may enhance return on asset investment. Decision-making uncertainty prompts customers to collaborate with service firms in making decisions. In other words, customers adjust their roles and are increasingly engaged in co-production when commitment, trust, asset specificity, and decision-making uncertainty are enhanced. Although studies have examined the preceding effects, to our best knowledge, none has empirically examined the simultaneous effects of all the curvilinear relationships in a single study. When these determinants are excessive, however, customers will not engage in co-production process. In brief, we suggest that the relationships of commitment, trust, asset specificity, and decision-making uncertainty with co-production are curvilinear or are inverse U-shaped. These new forms of curvilinear relationships have not been identified in existing literature on co-production; therefore, they complement extant linear approaches. Most importantly, we aim to consider both the bright and the dark sides of the determinants of co-production.

Keywords: co-production, commitment, trust, asset specificity, decision-making uncertainty

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Authors: Zeina Bou Diab, Arij Daou

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The neural mechanisms of sequential behaviors are intensively studied, with songbirds a focus for learned vocal production. We are studying the premotor nucleus HVC at a nexus of multiple pathways contributing to song learning and production. The HVC consists of multiple classes of neuronal populations, each has its own cellular, electrophysiological and functional properties. During singing, a large subset of motor cortex analog-projecting HVCRA neurons emit a single 6-10 ms burst of spikes at the same time during each rendition of song, a large subset of basal ganglia-projecting HVCX neurons fire 1 to 4 bursts that are similarly time locked to vocalizations, while HVCINT neurons fire tonically at average high frequency throughout song with prominent modulations whose timing in relation to song remains unresolved. This opens the opportunity to define models relating explicit HVC circuitry to how these neurons work cooperatively to control learning and singing. We developed conductance-based Hodgkin-Huxley models for the three classes of HVC neurons (based on the ion channels previously identified from in vitro recordings) and connected them in several physiologically realistic networks (based on the known synaptic connectivity and specific glutaminergic and gabaergic pharmacology) via different architecture patterning scenarios with the aim to replicate the in vivo firing patterning behaviors. We are able, through these networks, to reproduce the in vivo behavior of each class of HVC neurons, as shown by the experimental recordings. The different network architectures developed highlight different mechanisms that might be contributing to the propagation of sequential neural activity (continuous or punctate) in the HVC and to the distinctive firing patterns that each class exhibits during singing. Examples of such possible mechanisms include: 1) post-inhibitory rebound in HVCX and their population patterns during singing, 2) different subclasses of HVCINT interacting via inhibitory-inhibitory loops, 3) mono-synaptic HVCX to HVCRA excitatory connectivity, and 4) structured many-to-one inhibitory synapses from interneurons to projection neurons, and others. Replication is only a preliminary step that must be followed by model prediction and testing.

Keywords: computational modeling, neural networks, temporal neural sequences, ionic currents, songbird

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Authors: Vidya Badrinarayanan, Appa Iyer Sivakumar

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The coronavirus pandemic has sent an important message that future project teams need to be trained to work under virtual conditions, which has already become the new norm in organizations across the world. As organizations increasingly rely on virtual teams to achieve project objectives, it is essential to comprehend how leadership functions in virtual project teams. The purpose of this research is to analyze the leadership behaviors exhibited by project managers for building trust and managing conflicts effectively in virtual project teams. This convergent parallel mixed method research was conducted by surveying 185 virtual leaders and conducting a semi-structured interview with 13 senior virtual leaders involved in managing projects across the industry sectors. The research findings indicate that establishing trust and managing conflicts were ranked as significant challenges in leading virtual project teams in the post-pandemic era. In contrast to earlier findings, our research findings suggest that productivity was not ranked as a significant challenge in leading virtual project teams. This indeed is a positive finding for organizations to consider adopting virtual project teams in the long run. Additionally, the research findings recommend that virtual leaders need to strive to build a high-trust environment and develop effective conflict resolution skills to improve the effectiveness of virtual project teams. As the project management profession struggles with low project success rates, mixed-method research aims to contribute to the knowledge in the growing research area of virtual project leadership. This research contributes to the knowledge by offering first-person accounts from senior virtual leaders on the innovative strategies they had implemented for building trust and resolving conflicts effectively in the virtual project when there were limited opportunities for face-to-face interaction on account of the pandemic. In addition, the leadership framework created as a part of this research for trust development and conflict management in virtual project teams will guide project managers to improve virtual project team effectiveness.

Keywords: conflict management, trust building, virtual leadership, virtual teams

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Authors: Prabesh Karki, Shyam Thapa Chettri, Bajarang Prasad Sah, Manoj Bhattarai, Sudeep Mishra

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Introduction: Frontal sinus is frequently described as the most difficult sinus to access surgically due to its proximity to the cribriform plate, orbit, and anterior ethmoid artery. Frontal sinus surgery requires a detailed understanding of the cellular structure and FSDP unique to each patient, making high-resolution CT scans an indispensable tool to assess the difficulty of planned sinus surgery. International Frontal Sinus Anatomy Classification (IFAC) was developed to provide a more precise nomenclature for cells in the frontal recess, classifying cells based on their anatomic origin. Objectives: To assess the proportion of frontal cell variants defined by IFAC, variation with respect to age and gender. Methods: 54 cases were enrolled after a detailed clinical history, thorough general and physical examinations, and CT a report ordered in a film. Assessment and tabulation of the presence of frontal cells according to the IFAC analyzed. The prevalence of each cell type was calculated, and data were entered in MS Excel and analyzed using Statistical Package for the Social Sciences (SPSS). Descriptive statistics and frequencies were defined for categorical and numerical variables. Frequency, percentage, the mean and standard deviation were calculated. Result: Among 54 patients, 30 (55.6%) were male and 24 (44.4%) were female. The patient enrolled ranged from 18 to 78 years. Majority33.3% (n=18) were in age group of >50 years.According to IFAC, Agger nasi cells (92.6%) were most common, whereas supraorbital ethmoidal cells were least common 16 (29.6%). Prevalence of other frontoethmoidal cells was SAC- 57.4%, SAFC- 38.9%, SBC- 74.1%, SBFC- 33.3%, FSC- 38.9% of 54 cases. Conclusion: IFAC is an international consensus document that describes an anatomically precise nomenclature for classifying frontoethmoidal cells' anatomy. This study has defined the prevalence, symmetry and reliability of frontoethmoidal cells as established by the IFAC system as in other parts of the world.

Keywords: frontal sinus, frontoethmoidal cells, international frontal sinus anatomy classification

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Authors: Venuka Kuruwita Arachchige Don, Mohamed Shaheen, Chris Goodier

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Aerated autoclaved concrete (AAC) is known for its lightweight, easy handling, high thermal insulation, and extremely porous structure. Investigation of pore behavior in AAC is crucial for characterizing the material, standardizing design and production techniques, enhancing the mechanical, durability, and thermal performance, studying the effectiveness of protective measures, and analyzing the effects of weather conditions. The significant details of pores are complicated to observe with acknowledged accuracy. The High-resolution Field Emission Scanning Electron Microscope (FESEM) image analysis is a promising technique for investigating the pore behavior and density of AAC, which is adopted in this study. Mercury intrusion porosimeter and gas pycnometer were employed to characterize porosity distribution and density parameters. The analysis considered three different densities of AAC blocks and three layers in the altitude direction within each block. A set of understandings was presented to extract and analyze the details of pore shape, pore size, pore connectivity, and pore percentages from FESEM images of AAC. Average pore behavior outcomes per unit area were presented. Comparison of porosity distribution and density parameters revealed significant variations. FESEM imaging offered unparalleled insights into porosity behavior, surpassing the capabilities of other techniques. The analysis conducted from a multi-staged approach provides porosity percentage occupied by various pore categories, total porosity, variation of pore distribution compared to AAC densities and layers, number of two-dimensional and three-dimensional pores, variation of apparent and matrix densities concerning pore behaviors, variation of pore behavior with respect to aluminum content, and relationship among shape, diameter, connectivity, and percentage in each pore classification.

Keywords: autoclaved aerated concrete, density, imaging technique, microstructure, porosity behavior

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Authors: Haoxiang Desmond Lim, Jie Wu, Tze How Daniel New, Shengxian Shi

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This paper outlines the development of a novel experimental technique in quantifying supersonic jet flows, in an attempt to avoid seeding particle problems frequently associated with particle-image velocimetry (PIV) techniques at high Mach numbers. Based on optical flow algorithms, the idea behind the technique involves using high speed cameras to capture Schlieren images of the supersonic jet shear layers, before they are subjected to an adapted optical flow algorithm based on the Horn-Schnuck method to determine the associated flow fields. The proposed method is capable of offering full-field unsteady flow information with potentially higher accuracy and resolution than existing point-measurements or PIV techniques. Preliminary study via numerical simulations of a circular de Laval jet nozzle successfully reveals flow and shock structures typically associated with supersonic jet flows, which serve as useful data for subsequent validation of the optical flow based experimental results. For experimental technique, a Z-type Schlieren setup is proposed with supersonic jet operated in cold mode, stagnation pressure of 8.2 bar and exit velocity of Mach 1.5. High-speed single-frame or double-frame cameras are used to capture successive Schlieren images. As implementation of optical flow technique to supersonic flows remains rare, the current focus revolves around methodology validation through synthetic images. The results of validation test offers valuable insight into how the optical flow algorithm can be further improved to improve robustness and accuracy. Details of the methodology employed and challenges faced will be further elaborated in the final conference paper should the abstract be accepted. Despite these challenges however, this novel supersonic flow measurement technique may potentially offer a simpler way to identify and quantify the fine spatial structures within the shock shear layer.

Keywords: Schlieren, optical flow, supersonic jets, shock shear layer

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Authors: Tamar Trop, Boris Portnov

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Public Space Lighting (PSL) of outdoor urban areas promotes comfort, defines spaces and neighborhood identities, enhances perceived safety and security, and contributes to residential satisfaction and wellbeing. However, if excessive or misdirected, PSL leads to unnecessary energy waste and increased greenhouse gas emissions, poses a non-negligible threat to the nocturnal environment, and may become a potential health hazard. At present, PSL is designed according to international, regional, and national standards, which consolidate best practice. Yet, knowledge regarding the optimal light characteristics needed for creating a perception of personal comfort and safety in densely populated residential areas, and the factors associated with this perception, is still scarce. The presented study suggests a paradigm shift in designing PSL towards a user-centered approach, which incorporates pedestrians' perspectives into the process. The study is an ongoing joint research project between China and Israel Ministries of Science and Technology. Its main objectives are to reveal inhabitants' perceptions of and preferences for PSL in different densely populated neighborhoods in China and Israel, and to develop a model that links instrumentally measured parameters of PSL (e.g., intensity, spectra and glare) with its perceived comfort and quality, while controlling for three groups of attributes: locational, temporal, and individual. To investigate measured and perceived PSL, the study employed various research methods and data collection tools, developed a location-based mobile application, and used multiple data sources, such as satellite multi-spectral night-time light imagery, census statistics, and detailed planning schemes. One of the study’s preliminary findings is that higher sense of safety in the investigated neighborhoods is not associated with higher levels of light intensity. This implies potential for energy saving in brightly illuminated residential areas. Study findings might contribute to the design of a smart and adaptive PSL strategy that enhances pedestrians’ perceived safety and comfort while reducing light pollution and energy consumption.

Keywords: energy efficiency, light pollution, public space lighting, PSL, safety perceptions

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Authors: Teng Li, Kamran Mohseni

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This paper presents an inviscid regularization technique that is capable of regularizing the shocks and sharp interfaces simultaneously in the shock-interface interaction simulations. The direct numerical simulation of flows involving shocks has been investigated for many years and a lot of numerical methods were developed to capture the shocks. However, most of these methods rely on the numerical dissipation to regularize the shocks. Moreover, in high Reynolds number flows, the nonlinear terms in hyperbolic Partial Differential Equations (PDE) dominates, constantly generating small scale features. This makes direct numerical simulation of shocks even harder. The same difficulty happens in two-phase flow with sharp interfaces where the nonlinear terms in the governing equations keep sharpening the interfaces to discontinuities. The main idea of the proposed technique is to average out the small scales that is below the resolution (observable scale) of the computational grid by filtering the convective velocity in the nonlinear terms in the governing PDE. This technique is named “observable method” and it results in a set of hyperbolic equations called observable equations, namely, observable Navier-Stokes or Euler equations. The observable method has been applied to the flow simulations involving shocks, turbulence, and two-phase flows, and the results are promising. In the current paper, the observable method is examined on the performance of regularizing shocks and interfaces at the same time in shock-interface interaction problems. Bubble-shock interactions and Richtmyer-Meshkov instability are particularly chosen to be studied. Observable Euler equations will be numerically solved with pseudo-spectral discretization in space and third order Total Variation Diminishing (TVD) Runge Kutta method in time. Results are presented and compared with existing publications. The interface acceleration and deformation and shock reflection are particularly examined.

Keywords: compressible flow simulation, inviscid regularization, Richtmyer-Meshkov instability, shock-bubble interactions.

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Authors: Jeremy Johnson, Craig Nitschke, Luke Kelly

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Fire regimes, climate and topographic gradients interact to influence ecosystem structure and function across fire-prone, montane landscapes worldwide. Biota have developed a range of adaptations to historic fire regime thresholds, which allow them to persist in these environments. In south-eastern Australia, a signal of fire regime changes is emerging across these landscapes, and anthropogenic climate change is likely to be one of the main drivers of an increase in burnt area and more frequent wildfire over the last 25 years. This shift has the potential to modify vegetation structure and composition at broad scales, which may lead to landscape patterns to which biota are not adapted, increasing the likelihood of local extirpation of some mammal species. This study aimed to address concerns related to the influence of abrupt changes in fire regimes on mammal populations in montane landscapes. It first examined the impact of climate, topography, and vegetation on fire patterns and then explored the consequences of these changes on mammal populations and their habitats. Field studies were undertaken across diverse vegetation, fire severity and fire frequency gradients, utilising camera trapping and passive acoustic monitoring methodologies and the collection of fine-scale vegetation data. Results show that drought is a primary contributor to fire regime shifts at the landscape scale, while topographic factors have a variable influence on wildfire occurrence at finer scales. Frequent, high severity wildfire influenced forest structure and composition at broad spatial scales, and at fine scales, it reduced occurrence of hollow-bearing trees and promoted coarse woody debris. Mammals responded differently to shifts in forest structure and composition depending on their habitat requirements. This study highlights the complex interplay between fire regimes, environmental gradients, and biotic adaptations across temporal and spatial scales. It emphasizes the importance of understanding complex interactions to effectively manage fire-prone ecosystems in the face of climate change.

Keywords: fire, ecology, biodiversity, landscape ecology

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Authors: Juliana Bom-Tempo

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The proposed composition occurs between images, performances, clinics and philosophies. For this enterprise we depart for what is not known beforehand, so with a question as a compass: "would it be in the creation, production and implementation of images in a performance a 'when' for the event of a poetic clinic?” In light of this, there are, in order to think a 'when' of the event of a poetic clinic, images in performances created, produced and executed in partnerships with the author of this text. Faced with this composition, we built four indicators to find spatiotemporal coordinates that would spot that "when", namely: risk zones; the mobilizations of the signs; the figuring of the flesh and an education of the affections. We dealt with the images in performances; Crútero; Flesh; Karyogamy and the risk of abortion; Egg white; Egg-mouth; Islands, threads, words ... germs; Egg-Mouth-Debris, taken as case studies, by engendering risks areas to promote individuations, which never actualize thoroughly, thus always something of pre-individual and also individuating a environment; by mobilizing the signs territorialized by the ordinary, causing them to vary the language and the words of order dictated by the everyday in other compositions of sense, other machinations; by generating a figure of flesh, disarranging the bodies, isolating them in the production of a ground force that causes the body to leak out and undo the functionalities of the organs; and, finally, by producing an education of affections, by placing the perceptions in becoming and disconnecting the visible in the production of small deserts that call for the creation of a people yet to come. The performance is processed as a problematizing of the images fixed by the ordinary, producing gestures that precipitate the individuation of images in performance, strange to the configurations that gather bodies and spaces in what we call common. Lawrence proposes to think of "people" who continually use umbrellas to protect themselves from chaos. These have the function of wrapping up the chaos in visions that create houses, forms and stabilities; they paint a sky at the bottom of the umbrella, where people march and die. A chaos, where people live and wither. Pierce the umbrella for a desire of chaos; a poet puts himself as an enemy of the convention, to be able to have an image of chaos and a little sun that burns his skin. The images in performances presented, thereby, were moving in search for the power of producing a spatio-temporal "when" putting the territories in risk areas, mobilizing the signs that format the day-to-day, opening the bodies to a disorganization and the production of an education of affections for the event of a poetic clinic.

Keywords: Experimentations , Images in Performances, Poetic Clinic, Risk

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Authors: Mona Alharbi

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Melioidosis has emerged as a lethal disease. Unfortunately, the molecular mechanisms of virulence and pathogenicity of Burkholderia pseudomallei remain unknown. However, proteomics research has selected putative targets in B. pseudomallei that might play roles in the B. pseudomallei virulence. BPSL 2418 putative protein has been predicted as a free methionine sulfoxide reductase and interestingly there is a link between the level of the methionine sulfoxide in pathogen tissues and its virulence. Therefore in this work, we describe the cloning expression, purification, and crystallization of BPSL 2418 and the solution of its 3D structure using X-ray crystallography. Also, we aimed to identify the substrate binding and reduced forms of the enzyme to understand the role of BPSL 2418. The gene encoding BPSL2418 from B. pseudomallei was amplified by PCR and reclone in pETBlue-1 vector and transformed into E. coli Tuner DE3 pLacI. BPSL2418 was overexpressed using E. coli Tuner DE3 pLacI and induced by 300μM IPTG for 4h at 37°C. Then BPS2418 purified to better than 95% purity. The pure BPSL2418 was crystallized with PEG 4000 and PEG 6000 as precipitants in several conditions. Diffraction data were collected to 1.2Å resolution. The crystals belonged to space group P2 21 21 with unit-cell parameters a = 42.24Å, b = 53.48Å, c = 60.54Å, α=γ=β= 90Å. The BPSL2418 binding MES was solved by molecular replacement with the known structure 3ksf using PHASER program. The structure is composed of six antiparallel β-strands and four α-helices and two loops. BPSL2418 shows high homology with the GAF domain fRMsrs enzymes which suggest that BPSL2418 might act as methionine sulfoxide reductase. The amino acids alignment between the fRmsrs including BPSL 2418 shows that the three cysteines that thought to catalyze the reduction are fully conserved. BPSL 2418 contains the three conserved cysteines (Cys⁷⁵, Cys⁸⁵ and Cys¹⁰⁹). The active site contains the six antiparallel β-strands and two loops where the disulfide bond formed between Cys⁷⁵ and Cys¹⁰⁹. X-ray structure of free methionine sulfoxide binding and native forms of BPSL2418 were solved to increase the understanding of the BPSL2418 catalytic mechanism.

Keywords: X-Ray Crystallography, BPSL2418, Burkholderia pseudomallei, Melioidosis

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Authors: Qiaoyue Kuang, Yang Li, Mizuki Endo, Takeaki Ozawa

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G protein-coupled receptors (GPCR) are the largest family of receptor proteins that detect molecules outside the cell and activate cellular responses. Of the GPCRs, dopamine receptors, which recognize extracellular dopamine, are essential to mammals due to their roles in numerous physiological events, including autonomic movement, hormonal regulation, emotions, and the reward system in the brain. To precisely understand the physiological roles of dopamine receptors, it is important to spatiotemporally control the signaling mediated by dopamine receptors, which is strongly dependent on their surface expression. Conventionally, chemical-induced interactions were applied to trigger the endocytosis of cell surface receptors. However, these methods were subjected to diffusion and therefore lacked temporal and special precision. To further understand the receptor-mediated signaling and to control the plasma membrane expression of receptors, an optogenetic tool called E-fragment was developed. The C-terminus of a light-sensitive photosensory protein cyptochrome2 (CRY2) was attached to β-Arrestin, and the E-fragment was generated by fusing the C-terminal peptide of vasopressin receptor (V2R) to CRY2’s binding partner protein CIB. The CRY2-CIB heterodimerization triggered by blue light stimulation brings β-Arrestin to the vicinity of membrane receptors and results in receptor endocytosis. In this study, the E-fragment system was applied to dopamine receptors 1 and 2 (DRD1 and DRD2) to control dopamine signaling. First, confocal fluorescence microscope observation qualitatively confirmed the light-induced endocytosis of E-fragment fused receptors. Second, NanoBiT bioluminescence assay verified quantitatively that the surface amount of E-fragment labeled receptors decreased after light treatment. Finally, GloSensor bioluminescence assay results suggested that the E-fragment-dependent receptor light-induced endocytosis decreased cAMP production in DRD1 signaling and attenuated the inhibition effect of DRD2 on cAMP production. The developed optogenetic tool was able to induce receptor endocytosis by external light, providing opportunities to further understand numerous physiological activities by controlling receptor-mediated signaling spatiotemporally.

Keywords: dopamine receptors, endocytosis, G protein-coupled receptors, optogenetics

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