Search results for: multi-scale fusion

Authors: Carmela Perta

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Among the studies flourished in the field of historical sociolinguistics, mainly in the strand devoted to English history, during its Medieval and early modern phases, multilingual texts had been analysed using theories and models coming from contact linguistics, thus applying synchronic models and approaches to the past. This is true also in the case of contact phenomena which would transcend the writing level involving the language systems implicated in contact processes to the point of perceiving a new variety. This is the case for medieval administrative-commercial texts in which, according to some Scholars, the degree of fusion of Anglo-Norman, Latin and middle English is so high a mixed code emerges, and there are recurrent patterns of mixed forms. Interesting is a collection of multilingual business writings by John Balmayn, an Englishman overseeing a large shipment in Tuscany, namely the Cantelowe accounts. These documents display various analogies with multilingual texts written in England in the same period; in fact, the writer seems to make use of the above-mentioned patterns, with Middle English, Latin, Anglo-Norman, and the newly added Italian. Applying an atomistic yet dynamic approach to the study of contact phenomena, we will investigate these documents, trying to explore the nature of the switching forms they contain from an intra-writer variation perspective. After analysing the accounts and the type of multilingualism in them, we will take stock of the assumed mixed code nature, comparing the characteristics found in this genre with modern assumptions. The aim is to evaluate the possibility to consider the switching forms as core elements of a mixed code, used as professional variety among merchant communities, or whether such texts should be analysed from a switching perspective.

Keywords: historical sociolinguistics, historical code switching, letters, medieval england

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Authors: Matthew Ferguson, Tatyana Konkova, Ioannis Violatos

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Microstructure analysis of additively manufactured (AM) materials is an important step in understanding the interrelationship between mechanical properties and materials performance. Literature on the effect of laser-based AM process parameters on the microstructure in the substrate-deposit interface is limited. The interface region, the adjoining area of substrate and deposit, is characterized by the presence of the fusion zone (FZ) and heat-affected zone (HAZ), experiencing rapid thermal gyrations resulting in thermal-induced transformations. Inconel 718 was utilized as work material for both the substrate and deposit. Three blocks of Inconel 718 material were deposited by Direct Energy Deposition (DED) using three different laser powers, 550W, 750W and 950W, respectively. A coupled thermo-mechanical transient approach was utilized to correlate temperature history to the evolution of microstructure. The thermal history of the deposition process was monitored with the thermocouples installed inside the substrate material. The interface region of the blocks was analyzed with Optical Microscopy (OM) and Scanning Electron Microscopy (SEM), including the electron back-scattered diffraction (EBSD) technique. Laser power was found to influence the dissolution of intermetallic precipitated phases in the substrate and grain growth in the interface region. Microstructure and thermal history data were utilized to draw conclusive comparisons between the investigated process parameters.

Keywords: additive manufacturing, direct energy deposition, electron back-scattered diffraction, finite element analysis, inconel 718, microstructure, optical microscopy, scanning electron microscopy, substrate-deposit interface region

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Authors: Yaya Gao, Jifeng Liu, Yafeng Liu

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Objectives: This study aimed to improve clinicians' understanding and diagnosis of the Mammary analogue secretory carcinoma of the salivary gland（MASC）. Methods: The clinical features of a MASC patient who was admitted to WestChina Hospital of Sichuan University in July 2020 were reviewed and analyzed. A 49-year-old woman with left upper neck pain for three months was admitted to the hospital. She underwent adenoma resection of the left submandibular gland 14 years ago and mucoepidermoid carcinoma resection surgery five years ago. Three months before admission, the patient developed pain in the left mandibular angle after "fatigue" and gradually developed radiation pain in the left ear, which could be relieved after rest. A mass of 1cm could be touched at the mandibular, with tenderness, poor mobility, and hard texture. No swelling, heat, pain, rupture, or pus was found on the surrounding skin. Color doppler ultrasonography of the salivary gland indicated a weak echo mass of 23*14*17mm in the left parotid gland. Results: Surgical excision was completed. Immunohistochemistry of the tumor samples after operation showed that P63（a few，+）, CK7（+）, S100（+）, DOG1（-）, Ki67（MIB-1）（+，5%），pan-TRK（+）, PAS(+) . ETV-6 gene translocation was detected in FISH in postoperative pathology, which indicated MASC. After this diagnosis, the patient sent the postoperative specimen of the second submandibular tumor to our hospital for consultation. The morphology of the two was similar. FISH detected ETV-6 gene translocation, so the second pathological diagnosis was revised to MASC. Conclusion: MASC of the salivary gland is a rare salivary gland tumor whose diagnosis depends on the result of the ETV6-NTRK3 fusion gene.

Keywords: mammary analogue secretory carcinoma, ETV6-NTRK3, salivary gland, misdiagnosed

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Authors: Kuan-Hao Tsui, Li-Te Lin, Chia-Jung Li

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The combination of Y-27632 and Vitamin C significantly enhances the quality of aging germ cells by reducing reactive oxygen species (ROS) production, restoring mitochondrial membrane potential balance, and promoting mitochondrial fusion. The age-related decline in oocyte quality contributes to reduced fertility, increased aneuploidy, and diminished embryo quality, with mitochondrial dysfunction in both oocytes and granulosa cells being a key factor in this decline. Experiments on aging germ cells investigated the effects of the Y-27632 and Vitamin C combination. In vivo studies involved aged mice to assess oocyte maturation and ROS accumulation during culture. The assessment included mitochondrial activity, ROS levels, mitochondrial membrane potential, and mitochondrial dynamics. Cellular energy metabolism and ATP production were also measured. The combination treatment effectively addressed mitochondrial dysfunction and regulated cellular energy metabolism, promoting oxygen respiration and increasing ATP production. In aged mice, this supplement treatment enhanced in vitro oocyte maturation and prevented ROS accumulation in aging oocytes during culture. While these findings are promising, further research is needed to explore the long-term effects and potential side effects of the Y-27632 and Vitamin C combination. Additionally, translating these findings to human subjects requires careful consideration. Overall, the study suggests that the Y-27632 and Vitamin C combination could be a promising intervention to mitigate aging-related dysfunction in germ cells, potentially enhancing oocyte quality, particularly in the context of in vitro fertilization.

Keywords: ovarian aging, supplements, ROS, mitochondria

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Authors: Lanchi Xie, Zhihui Li, Zhigang Li, Guiqiang Wang, Lei Xu, Yuwen Yan

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Image-based facial features can be classified into category recognition features and individual recognition features. Current automated face recognition systems extract a specific feature vector of different dimensions from a facial image according to their pre-trained neural network. However, to improve the efficiency of parameter calculation, an algorithm generally reduces the image details by pooling. The operation will overlook the details concerned much by forensic experts. In our experiment, we adopted a variety of face recognition algorithms based on deep learning, compared a large number of naturally collected face images with the known data of the same person's frontal ID photos. Downscaling and manual handling were performed on the testing images. The results supported that the facial recognition algorithms based on deep learning detected structural and morphological information and rarely focused on specific markers such as stains and moles. Overall performance, distribution of genuine scores and impostor scores, and likelihood ratios were tested to evaluate the accuracy of biometric systems and forensic experts. Experiments showed that the biometric systems were skilled in distinguishing category features, and forensic experts were better at discovering the individual features of human faces. In the proposed approach, a fusion was performed at the score level. At the specified false accept rate, the framework achieved a lower false reject rate. This paper contributes to improving the interpretability of the objective method of facial comparison and provides a novel method for human-machine collaboration in this field.

Keywords: likelihood ratio, automated facial recognition, facial comparison, biometrics

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Authors: Gheorghe Ionita, Sebastian Brad, Ioan Stefanescu

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In any fusion device, tritium plays a key role both as a fuel component and, due to its radioactivity and easy incorporation, as tritiated water (HTO). As for the ITER project, to reduce the constant potential of tritium emission, there will be implemented a Water Detritiation System (WDS) and an Isotopic Separation System (ISS). In the same time, during operation of fission CANDU reactors, the tritium content increases in the heavy water used as moderator and cooling agent (due to neutron activation) and it has to be reduced, too. In Romania, at the National Institute for Cryogenics and Isotopic Technologies (ICIT Rm-Valcea), there is an Experimental Pilot Plant for Tritium Removal (Exp. TRF), with the aim of providing technical data on the design and operation of an industrial plant for heavy water depreciation of CANDU reactors from Cernavoda NPP. The selected technology is based on the catalyzed isotopic exchange process between deuterium and liquid water (LPCE) combined with the cryogenic distillation process (CD). This paper presents an updated review of activities in the field carried out in Romania after the year 2000 and in particular those related to the development and operation of Tritium Removal Experimental Pilot Plant. It is also presented a comparison between the experimental pilot plant and industrial plant to be implemented at Cernavoda NPP. The similarities between the experimental pilot plant from ICIT Rm-Valcea and water depreciation and isotopic separation systems from ITER are also presented and discussed. Many aspects or 'opened issues' relating to WDS and ISS could be checked and clarified by a special research program, developed within ExpTRF. By these achievements and results, ICIT Rm - Valcea has proved its expertise and capability concerning tritium management therefore its competence may be used within ITER project.

Keywords: ITER project, heavy water detritiation, tritium removal, isotopic exchange

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Authors: Benedict O. Muyale, Emmanuel S. Murunga

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Cities have always been the loci for nationals as well as growth of cultural fusion and innovation. Over 50%of global population dwells in cities and urban centers. This means that cities are prolific users of natural resources and generators of waste; hence they produce most of the greenhouse gases which are causing global climate change. The root cause of increase in the transport sector carbon curve is mainly the greater numbers of individually owned cars. Development in these cities is geared towards economic progress while environmental sustainability is ignored. Infrastructure projects focus on road expansion, electrification, and more parking spaces. These lead to more carbon emissions, traffic congestion, and air pollution. Recent development plans for Nairobi city are now on road expansion with little priority for electric train solutions. The Vision 2030, Kenya’s development guide, has shed some light on the city with numerous road expansion projects. This chapter seeks to realize the following objectives; (1) to assess the current transport situation of Nairobi; (2) to review green transport solutions being undertaken in the city; (3) to give an overview of alternative green transportation solutions, and (4) to provide a green transportation framework matrix. This preliminary study will utilize primary and secondary data through mainly desktop research and analysis, literature, books, magazines and on-line information. This forms the basis for formulation of approaches for incorporation into the green transportation framework matrix of the main study report.The main goal is the achievement of a practical green transportation system for implementation by the City County of Nairobi to reduce carbon emissions and congestion and promote environmental sustainability.

Keywords: cities, transport, Nairobi, green technologies

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Authors: Ximena Calle, Plinio Cantero-López, Felipe Muñoz-Córdova, Mayarling-Francisca Troncoso, Sergio Lavandero, Valentina Parra

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MUL1, a mitochondrial E3 ubiquitin ligase anchored to the outer mitochondrial membrane, is highly expressed in the heart. MUL1 is involved in multiple biological pathways associated with mitochondrial dynamics. Increased MUL1 affects the balance between fission and fusion, affecting mitochondrial function, which plays a crucial role in myocardial function. Therefore, it is interesting to evaluate the effect of cardiac-specific overexpression of MUL1 on myocardial function. Aim: To determine heart functionality in a mouse model with cardio-specific overexpression MUL1 protein. Methods and Results: Male C57BL/Tg transgenic mice with cardiomyocyte-specific overexpression of MUL1 (n=10) and control (n=4) were evaluated at 12, 27, and 35 weeks of age. Glucose tolerance curve determination was performed after a 6-hours fast to assess metabolic capacity, treadmill test, and systolic, and diastolic pressure was evaluated by the mouse tail-cuff blood pressure system equipment. The result showed no glucose tolerance curve, and the treadmill test demonstrated no significant changes between groups. However, substantial changes in diastolic function were observed by ultrasound and determination of cardiac hypertrophy proteins by western blot. Conclusions: Cardio-specific overexpression of MUL1 in mice without any treatment affects diastolic cardiac function, thus showing the important role contributed by MUL1 in the heart. Future research should evaluate the effect of cardiomyocyte-specific overexpression of MUL1 in pathological conditions such as a high-fat diet is one of the main risk factors for cardiovascular disease.

Keywords: diastolic dysfunction, hypertrophy cardiac, mitochondrial E3 ubiquitin ligase 1, MUL1

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Authors: Ilja Käch, Abdul R. Jandali, Nadja Zechmann-Müller

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Introduction: We discuss the treatment of two young male patients suffering from scaphoid non-union after a traumatic scaphoid fracture. The currently propagated techniques for treating a scaphoid non-union in children are either the operative reconstruction of the scaphoid or the conservative treatment with splinting in a scaphoid cast. Cases: In the first case, we operated on a 13 years old male patient with a posttraumatic scaphoid non-union in the middle third with a humpback deformity. We resected the middle third of the scaphoid and grafted the defect with an iliac crest bone, and the DISI-Deformity was reduced. Fixation was performed with K-Wires and immobilisation in a scaphoid cast. In the second case a 13 years old male patient also with a posttraumatic scaphoid non-union in the middle third and humpback deformity, DISI-deformity, was treated conservatively. Immobilisation in a scaphoid cast for four months was performed. Results: Operative: One year postoperatively the patient achieved a painless free arc of motion. Flexion/Extension 70/0/60°, Radial-/Ulnarduction 30/0/30° and Pro-/Supination 90/0/90°. The computer tomogram showed complete consolidation and bony fusion of the iliac crest bone. Conservative: Six to eight months after conservative treatment the patient demonstrated painless motion and AROM Flexion/Extension 80/0/80°, Radial-/Ulnarduction and Pro-/Supination in maximum range. Complete consolidation in the computer tomogram with persistent humpback- and DISI deformity. Conclusion: In the literature, both techniques are described, either the operative scaphoid reconstruction or the conservative treatment with splinting. In our cases, both the operative and conservative treatments showed comparable good results. However, the humpback- and DISI deformity can only be addressed with a surgical approach.

Keywords: scaphoid, non-union, trauma, operative vs. non operative

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Authors: Svanik Garg

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Static Test Pads are stationary mechanisms that hold a solid rocket motor, measuring the different parameters of its operation including thrust and temperature to better calibrate it for launch. This paper outlines a specific STP designed to test high powered rocket motors with a thrust upwards of 4000N and limited to 6500N. The design includes a specific portable mechanism with cost an integral part of the design process to make it accessible to small scale rocket developers with limited resources. Using curved surfaces and an ergonomic design, the STP has a delicately engineered façade/case with a focus on stability and axial calibration of thrust. This paper describes the design, operation and working of the STP and its widescale uses given the growing market of aviation enthusiasts. Simulations on the CAD model in Fusion 360 provided promising results with a safety factor of 2 established and stress limited along with the load coefficient A PCB was also designed as part of the test pad design process to help obtain results, with visual output and various virtual terminals to collect data of different parameters. The circuitry was simulated using ‘proteus’ and a special virtual interface with auditory commands was also created for accessibility and wide-scale implementation. Along with this description of the design, the paper also emphasizes the design principle behind the STP including a description of its vertical orientation to maximize thrust accuracy along with a stable base to prevent micromovements. Given the rise of students and professionals alike building high powered rockets, the STP described in this paper is an appropriate option, with limited cost, portability, accuracy, and versatility. There are two types of STP’s vertical or horizontal, the one discussed in this paper is vertical to utilize the axial component of thrust.

Keywords: static test pad, rocket motor, thrust, load, circuit, avionics, drag

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Authors: Yu-Chieh Chen, Li-Yun Wang, Chi-Chih Chen, Huy Hùng Đào, Ya-Mei Chen, Ming-Chu Cheng, Wen-Bin Chung, Hso-Chi Chaung, Guan-Ming Ke

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Many recent researches have demonstrated that CD154, a protein primarily expressed on activated T cell molecules, has potentially acted as a molecular adjuvant to improve the immunogenicity of subunit vaccines against viral infections. Classical swine fever (CSF) affects the swine industry worldwide that is one of the most devastating and highly contagious pig diseases. It is listed by the World Organization for Animal Health (OIE) as an infectious animal disease that must be reported. Although pigs vaccinated with subunit vaccines can be differentially diagnosed from those infected animals, subunit vaccines usually need adjuvants to enhance and elicit immune responses. In this study, CD154 was linked with CSFV E2 sequences and then expressed in CHO cells to produce the fusion protein as E2-CD154. The porcine specific CpG adjuvant was also used in one of the formulations. The specific pathogen-free pigs (SPF) at the age of 4-week-old were randomly separated into four groups, vaccinated with E2-CpG, E2-CD154, E2-CD154-CpG or the commercial Bayovac® CSF-E2 vaccine and boosted two weeks after primary vaccination. The results showed that the percentages of CD4+ and CD4+IL2+ in peripheral blood mononuclear cells (PBMC) in E2-CD154 vaccinated piglets seven days after primary vaccination were gained by 1-5% relative to the control group. In addition, the percentages of CD4+IFNγ+ T cells had slightly edged up 0.1-0.3% compared with the control group. Also, increased E2-specific IFNγ levels had edged up CD4+CD8+ T cells found in E2-CD154 and E2-CD154-CpG groups, particularly in the E2-CD154-CpG group. These results implicate that CD154 may enhance cellular immunity and synergistically act with species-specific CpG adjuvant as a dual-phase adjuvant. Therefore, the CD154 may be beneficial as a promising adjuvant in subunit vaccines.

Keywords: CD154, CpG adjuvant, cellular immunity, subunit vaccine, pig

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Authors: Sadman Durlov, Aditya Ganesh-Ram, Hamidreza Hekmatjou, Md Najmus Salehin, Nora Shayesteh Ameri

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In the field of additive manufacturing, tungsten stands out for its exceptional resistance to high temperatures, making it an ideal candidate for use in extreme conditions. However, its inherent brittleness and vulnerability to thermal cracking pose significant challenges to its manufacturability. This study explores the microstructural evolution of tungsten processed through layer-wise rolling in laser powder bed fusion additive manufacturing, utilizing a comprehensive approach that combines advanced simulation techniques with empirical research. We aim to uncover the complex processes of plastic deformation and microstructural transformations, with a particular focus on the dynamics of grain size, boundary evolution, and phase distribution. Our methodology employs a combination of simulation and experimental data, allowing for a detailed comparison that elucidates the key mechanisms influencing microstructural alterations during the rolling process. This approach facilitates a deeper understanding of the material's behavior under additive manufacturing conditions, specifically in terms of deformation and recrystallization. The insights derived from this research not only deepen our theoretical knowledge but also provide actionable strategies for refining manufacturing parameters to improve the tungsten components' mechanical properties and functional performance. By integrating simulation with practical experimentation, this study significantly enhances the field of materials science, offering a robust framework for the development of durable materials suited for challenging operational environments. Our findings pave the way for optimizing additive manufacturing techniques and expanding the use of tungsten across various demanding sectors.

Keywords: additive manufacturing, layer wise rolling, refractory materials, in-situ microstructure modifications

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Authors: Gennaro Marino

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This paper concerns a case of a 10.75 inch diameter buried gas transmission line which was exposed to mine subsidence ground movements. The pipeline was buried about 4ft. below the surface with maximum operating pressure of 1440 psi. The mine subsidence movement was the result of long walling ore at a depth of approximately 1600 ft. As ore extraction progressed, the stress in the monitored pipeline worsened and was approaching unacceptable levels. The excessive pipe compression resulted when it was exposed to the compression zone of subsidence basin created by mining. The pipe stress reached a significant compressive level due to the extensive length of the pipe exposed to frictional ground-pipe slip resistance. The backfill ground movement slip resistance depends on normal stress around the pipe, the rate of slip, and the backfill characteristics. Normal stress depends on the burial depth of the backfill density and the lateral subsidence induced stress. The backfill in this site has a soil dry density of approximately 90 PCF. A suite of direct shear tests was conducted a residual friction angle of 36 was determined for the ambient backfill. These tests showed that the residual shearing resistance was reached within a fraction of an inch. The pipe was coated with fusion-bonded epoxy, so friction reduce factory of 0.6 can be considered. To relieve ground movement induced compressive stress, the line was uncovered. As more of the pipeline was exposed, the pipe abruptly bowed in the excavation. An analysis of this pipe formation which was performed is provided in this paper. Also discussed in this paper are ways to mitigate this pipe deformation or upheaval buckling from occurring. Keywords: Pipe Upheaval, Pipe Buckling, Ground subsidence, Buried Pipeline, Pipe Stress Mitigation.

Keywords: pipe upheaval, pipe buckling, ground subsidence, buried pipeline, pipe stress mitigation

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Authors: Dario Milani, Guido Morgenthal

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Fluid dynamic computation of wind caused forces on bluff bodies e.g light flexible civil structures or high incidence of ground approaching airplane wings, is one of the major criteria governing their design. For such structures a significant dynamic response may result, requiring the usage of small scale devices as guide-vanes in bridge design to control these effects. The focus of this paper is on the numerical simulation of the bluff body problem involving multiscale phenomena induced by small scale devices. One of the solution methods for the CFD simulation that is relatively successful in this class of applications is the Vortex Particle Method (VPM). The method is based on a grid free Lagrangian formulation of the Navier-Stokes equations, where the velocity field is modeled by particles representing local vorticity. These vortices are being convected due to the free stream velocity as well as diffused. This representation yields the main advantages of low numerical diffusion, compact discretization as the vorticity is strongly localized, implicitly accounting for the free-space boundary conditions typical for this class of FSI problems, and a natural representation of the vortex creation process inherent in bluff body flows. When the particle resolution reaches the Kolmogorov dissipation length, the method becomes a Direct Numerical Simulation (DNS). However, it is crucial to note that any solution method aims at balancing the computational cost against the accuracy achievable. In the classical VPM method, if the fluid domain is discretized by Np particles, the computational cost is O(Np2). For the coupled FSI problem of interest, for example large structures such as long-span bridges, the aerodynamic behavior may be influenced or even dominated by small structural details such as barriers, handrails or fairings. For such geometrically complex and dimensionally large structures, resolving the complete domain with the conventional VPM particle discretization might become prohibitively expensive to compute even for moderate numbers of particles. It is possible to reduce this cost either by reducing the number of particles or by controlling its local distribution. It is also possible to increase the accuracy of the solution without increasing substantially the global computational cost by computing a correction of the particle-particle interaction in some regions of interest. In this paper different strategies are presented in order to extend the conventional VPM method to reduce the computational cost whilst resolving the required details of the flow. The methods include temporal sub stepping to increase the accuracy of the particles convection in certain regions as well as dynamically re-discretizing the particle map to locally control the global and the local amount of particles. Finally, these methods will be applied on a test case and the improvements in the efficiency as well as the accuracy of the proposed extension to the method are presented. The important benefits in terms of accuracy and computational cost of the combination of these methods will be thus presented as long as their relevant applications.

Keywords: adaptation, fluid dynamic, remeshing, substepping, vortex particle method

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Authors: Sara Franco Ortega, Alina Goldberg Cavalleri, Nawaporn Onkokesung, Richard Dale, Melissa Brazier-Hicks, Robert Edwards

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Safeners are agrochemicals that enhance the selective chemical control of wild grasses by increasing the ability of the crop to metabolise the herbicide. Although these compounds are widely used, their mode of action is not well understood. It is known that safeners enhance the metabolism of herbicides, by up-regulating the associated detoxification system we have termed the xenome. The xenome proteins involved in herbicide metabolism have been previously divided into four different phases, with cytochrome P450s (CYPs) playing a key role in phase I metabolism by catalysing hydroxylation and dealkylation reactions. Subsequently, glutathione S-transferases (GSTs) and UDP-glucosyltransferases lead to the formation of Phase II conjugates prior to their transport into the vacuole by ABCs transporters (Phase III). Maize (Zea mays), was been treated with different safeners to explore the selective induction of xenome proteins, with a special interest in the regulation of the CYP superfamily. Transcriptome analysis enabled the identification of key safener-inducible CYPs that were then functionally assessed to determine their role in herbicide detoxification. In order to do that, CYP’s were codon optimised, synthesised and inserted into the yeast expression vector pYES3 using in-fusion cloning. CYP’s expressed as recombinant proteins in a strain of yeast engineered to contain the P450 co-enzyme (cytochrome P450 reductase) from Arabidopsis. Microsomes were extracted and treated with herbicides of different chemical classes in the presence of the cofactor NADPH. The reaction products were then analysed by LCMS to identify any herbicide metabolites. The results of these studies will be presented with the key CYPs identified in maize used as the starting point to find orthologs in other crops and weeds to better understand their roles in herbicide selectivity and safening.

Keywords: CYPs, herbicide detoxification, LCMS, RNA-Seq, safeners

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Authors: Dileep K. Verma, Sunil K. Lal

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Influenza still remains one of the most challenging diseases posing significant threat to public health causing seasonal epidemics and pandemics. Previous studies suggest that influenza hemagglutinin is essential for viral attachment to host sialic acid receptors and concentrate in lipid rafts for efficient viral fusion. Studies also reported selective nature of Influenza virus to utilize rafts micro-domain for efficient virus assembly and budding. However, the detailed mechanism of Influenza A Virus (IAV) binding to host cell membrane and entry inside the host remains elusive. In the present study, we investigated if host membrane lipid rafts play any significant role in early life cycle events of influenza A virus. Role of host lipid rafts was studied using raft disruption method by extraction of cholesterol and Methyl-β-Cyclodextrin was used to remove membrane cholesterol. We observed co-localization of Influenza A Virus to lipid rafts by visualization of known lipid raft marker GM1 on host cell membrane. Co-localization suggest direct involvement of these micro-domain in initiation of IAV life cycle. We found significant reduction in influenza A virus adsorption in raft disrupted target host cells indicating poor binding and attachment in absence of coherent membrane rafts. Taken together, the results of present study provide evidence for critical involvement of host lipid rafts and its constituents in adsorption process of Influenza A Virus and suggests crucial involvement in other early events of IAV life cycle. The present study opens a new domain to study influenza virus-host interaction and to combat flu at the very early steps of viral life cycle.

Keywords: lipid raft, adsorption, cholesterol, methyl-β-cyclodextrin, GM1

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Authors: Suneeta Gireesh Panicker

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Aim: Aminopeptidase P (APP) is an enzyme that has specificity for proline, can specifically cleave Xaa-Proline peptides and is a metallo-aminopeptidase. The bonds nearby to the imino acid proline are tough to cleave by many peptidases, but APP can specifically break peptide bonds engaged with proline. Membrane-bound form and a cytosolic form are the two forms in which this enzyme exists. The exact physiological function of APP remains unclear and hence the present work attempts to determine it. Methods: In the present study, the expression pattern of cytosolic Aminopeptidase P (DAP) was determined in all the embryonic stages and larval stages of wild-type Drosophila by using polyclonal monospecific antibodies. To show the presence of DAP RNA in embryonic and larval stages, RNA in situ hybridization was performed. DAP promoter-LacZ fusion reporter gene vector was used to construct transgenic embryos to study the regulation pattern of DAP. To study the DAP expression profile, a transgenic fly consisting of a DAP promoter with β-gal and GFP reporter genes in front of it was constructed. Results: DAP protein expression was observed in neuroectodermal cells, posterior midgut primordium, proctodeum, ventral neuroblast and primordial stomatogastric nervous system. It was observed in the ventral cord and midgut in stage 12. The completely developed embryos showed the intense occurrence of it in the ventral cord and gut region. The eye-antennal disc, wing disc and leg disc also showed the presence of DAP protein. LacZ expression in transgenic embryos also showed the same pattern. Conclusion: Similar to various known multiple-functional proteins, DAP could be one with different functions at different stages and in different cells. Data presented here designates DAP functions in the early embryonic and imaginal dics differentiation and development, suggesting that it may be required for the metabolism of proteins like neuropeptides and tachykinins.

Keywords: aminopeptidase P, in situ hybridization, transgenic fly, embryonic stages

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Authors: Indu ray

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In today’s digital world the internet is already flooded with information on how to master grammar in a foreign language. It is well known that one cannot master a language without grammar. Grammar is the backbone of any language. Without grammar there would be no structure to help you speak/write or listen/read. Successful communication is only possible if the form and function of linguistic utterances are firmly related to one another. Grammar has its own rules of use to formulate an easier-to-understand language. Like a tool, grammar formulates our thoughts and knowledge in a meaningful way. Every language has its own grammar. With grammar, we can quickly analyze whether there is any action in this text: (Present, past, future). Knowledge of grammar is an important prerequisite for mastering a foreign language. What’s most important is how teachers can make grammar lessons more interesting for students and thus promote grammar skills more successfully. Through this paper, we discuss a few important methods like (Interactive Grammar Exercises between students, Interactive Grammar Exercise between student to teacher, Grammar translation method, Audio -Visual Method, Deductive Method, Inductive Method). This paper is divided into two sections. In the first part, brief definitions and principles of these approaches will be provided. Then the possibility and the case of combination of this approach will be analyzed. In the last section of the paper, I would like to present a survey result conducted at my university on a few methods to quickly learn grammar in Foreign Language. We divided the Grammatical Skills in six Parts. 1.Grammatical Competence 2. Speaking Skills 3. Phonology 4. The syntax and the Semantics 5. Rule 6. Cognitive Function and conducted a survey among students. From our survey results, we can observe that phonology, speaking ability, syntax and semantics can be improved by inductive method, Audio-visual Method, and grammatical translation method, for grammar rules and cognitive functions we should choose IGE (teacher-student) method. and the IGE method (pupil-pupil). The study’s findings revealed, that the teacher delivery Methods should be blend or fusion based on the content of the Grammar.

Keywords: innovative method, grammatical skills, audio-visual, translation

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Authors: Benjamin Obeghare Izu

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In Africa, indigenous musical practices are the focal point in which most cultural practices revolve, and they are the conduit mainly used in transmitting Indigenous knowledge and values. They serve as a means of documenting, preserving, transmitting indigenous knowledge, and re-enacting their historical, social, and cultural affinity. Indigenous musical practices also serve as a repository for indigenous knowledge and artistic traditions. However, these indigenous musical practices and the resulting cultural ideals are confronted with substantial challenges in the twenty-first century from contemporary cultural influence. Additionally, indigenous musical practices' educational and cultural purposes have been impacted by the broad monetisation of the arts in contemporary society. They are seen as objects of entertainment. Some young people are today unaware of their cultural roots and are losing their cultural identity due to these influences and challenges. In order to help policymakers raise awareness of and encourage the use of indigenous knowledge and musical practices among African youth and scholars, this study is in response to the need to explore the components and functions of the indigenous knowledge system, values, and musical tradition in Africa. The study employed qualitative research methods, utilising interviews, participant observation, and conducting related literature as data collection methods. It examines the indigenous musical practices in the Oba of Benin Royal Igue festival among the Benin people in Edo state, Nigeria, and the Ovwuwve festival observed by the Abraka people in Delta state, Nigeria. The extent to which the indigenous musical practices convey and protect indigenous knowledge and cultural values are reflected in the musical practices of the cultural festivals. The study looks at how indigenous musical arts are related to one another and how that affects how indigenous knowledge is transmitted and preserved. It makes recommendations for how to increase the use of indigenous knowledge and values and their fusion with contemporary culture. The study contributes significantly to ethnomusicology by showing how African traditional music traditions support other facets of culture and how indigenous knowledge might be helpful in contemporary society.

Keywords: African musical practices, African music and dance, African society, indigenous musical practices

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Authors: Seyyed Mohammad Amin Mousavi Sagharchi, Elham Javanroudi

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Introduction: Tuberculosis (TB) is a known disease with hidden features caused by Mycobacterium tuberculosis (MTB). This disease, which is one of the 10 deadliest in the world, has caused millions of deaths in recent decades. Furthermore, TB is responsible for infecting about 30% population of world. Like any infection, TB can activate the immune system by locating and colonization in the human body, especially in the alveoli. TB is granulomatosis, so MTB can absorb the host’s immune cells and other cells to form granuloma. Method: Different databases (e.g., PubMed) were recruited to prepare this paper and fulfill our goals to search and find effective papers and investigations. Results: Immune response to MTB is related to T cell killers and contains CD1, CD4, and CD8 T lymphocytes. CD1 lymphocytes can recognize glycolipids, which highly exist in the Mycobacterial fatty cell wall. CD4 lymphocytes and macrophages form granuloma, and it is the main line of immune response to Mycobacteria. On the other hand, CD8 cells have cytolytic function for directly killing MTB by secretion of granulysin. Other functions and secretion to the deal are interleukin-12 (IL-12) by induction of expression interferon-γ (INF-γ) for macrophages activation and creating a granuloma, and tumor necrosis factor (TNF) by promoting macrophage phagolysosomal fusion. Conclusion: Immune cells in battle with MTB are macrophages, dendritic cells (DCs), neutrophils, and natural killer (NK) cells. These immune cells can recognize the Mycobacterium by various receptors, including Toll-like receptors (TLRs), Nod-like receptors (NLRs), and C-type lectin receptors (CLRs) located in the cell surface. In human alveoli exist about 50 dendritic macrophages, which have close communication with other immune cells in the circulating system and epithelial cells to deal with Mycobacteria. Against immune cells, MTB handles some factors (e.g., cordfactor, O-Ag, lipoarabinomannan, sulfatides, and adenylate cyclase) and practical functions (e.g., inhibition of macrophages).

Keywords: mycobacterium tuberculosis, immune responses, immunological mechanisms, respiratory tuberculosis

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Authors: Monish Sengupta, Benjamin Heydecker, Daniel Woodland

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New train control systems rely mainly on Automatic Train Protection (ATP) and Automatic Train Operation (ATO) dynamically to control the speed and hence performance. The ATP and the ATO form the vital element within the CBTC (Communication Based Train Control) and within the ERTMS (European Rail Traffic Management System) system architectures. Reliable and accurate measurement of train location, speed and acceleration are vital to the operation of train control systems. In the past, all CBTC and ERTMS system have deployed a balise or equivalent to correct the uncertainty element of the train location. Typically a CBTC train is allowed to miss only one balise on the track, after which the Automatic Train Protection (ATP) system applies emergency brake to halt the service. This is because the location uncertainty, which grows within the train control system, cannot tolerate missing more than one balise. Balises contribute a significant amount towards wayside maintenance and studies have shown that balises on the track also forms a constraint for future track layout change and change in speed profile.This paper investigates the causes of the location uncertainty that is currently experienced and considers whether it is possible to identify an effective filter to ascertain, in conjunction with appropriate sensors, more accurate speed, distance and location for a CBTC driven train without the need of any external balises. An appropriate sensor fusion algorithm and intelligent sensor selection methodology will be deployed to ascertain the railway location and speed measurement at its highest precision. Similar techniques are already in use in aviation, satellite, submarine and other navigation systems. Developing a model for the speed control and the use of Kalman filter is a key element in this research. This paper will summarize the research undertaken and its significant findings, highlighting the potential for introducing alternative approaches to train positioning that would enable removal of all trackside location correction balises, leading to huge reduction in maintenances and more flexibility in future track design.

Keywords: ERTMS, CBTC, ATP, ATO

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Authors: Chuyuan Wang, Nayan Khare, Lily Villa, Patricia Solis, Elizabeth A. Wentz

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Maricopa County, Arizona, has a semi-arid hot desert climate that is one of the hottest regions in the United States. The exacerbated urban heat island (UHI) effect caused by rapid urbanization has made the urban area even hotter than the rural surroundings. The Phoenix metropolitan area experiences extremely high temperatures in the summer from June to September that can reach the daily highest of 120 °F (48.9 °C). Morbidity and mortality due to the environmental heat is, therefore, a significant public health issue in Maricopa County, especially because it is largely preventable. Public records from the Maricopa County Department of Public Health (MCDPH) revealed that between 2012 and 2016, there were 10,825 incidents of heat-related morbidity incidents, 267 outdoor environmental heat deaths, and 173 indoor heat-related deaths. A lot of research has examined heat-related death and its contributing factors around the world, but little has been done regarding heat-related morbidity issues, especially for regions that are naturally hot in the summer. The objective of this study is to examine the demographic, socio-economic, housing, and environmental factors that contribute to heat-related morbidity in Maricopa County. We obtained heat-related morbidity data between 2012 and 2016 at census tract level from MCDPH. Demographic, socio-economic, and housing variables were derived using 2012-2016 American Community Survey 5-year estimate from the U.S. Census. Remotely sensed Landsat 7 ETM+ and Landsat 8 OLI satellite images and Level-1 products were acquired for all the summer months (June to September) from 2012 and 2016. The National Land Cover Database (NLCD) 2016 percent tree canopy and percent developed imperviousness data were obtained from the U.S. Geological Survey (USGS). We used ordinary least squares (OLS) regression analysis to examine the empirical relationship between all the independent variables and heat-related morbidity rate. Results showed that higher morbidity rates are found in census tracts with higher values in population aged 65 and older, population under poverty, disability, no vehicle ownership, white non-Hispanic, population with less than high school degree, land surface temperature, and surface reflectance, but lower values in normalized difference vegetation index (NDVI) and housing occupancy. The regression model can be used to explain up to 59.4% of total variation of heat-related morbidity in Maricopa County. The multiscale geographically weighted regression (MGWR) technique was then used to examine the spatially varying relationships between heat-related morbidity rate and all the significant independent variables. The R-squared value of the MGWR model increased to 0.691, that shows a significant improvement in goodness-of-fit than the global OLS model, which means that spatial heterogeneity of some independent variables is another important factor that influences the relationship with heat-related morbidity in Maricopa County. Among these variables, population aged 65 and older, the Hispanic population, disability, vehicle ownership, and housing occupancy have much stronger local effects than other variables.

Keywords: census, empirical modeling, heat-related morbidity, spatial analysis

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Authors: Mahmoud A. Amin, Essam Taman, Ahmed Omran, Mahmoud Shawky, Ahmed Mekawy, Abdallah M. Kotkat, Saber Younes, Nehad N. Ghonemy, Amin Saad, Ezz-Aleslam, Abdullah M. Elosh

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Introduction: TMJ is a one-of-a-kind, complicated synovial joint that helps with masticatory function by allowing the mandible to open and close the mouth. True ankylosis is a situation in which condylar movement is limited by a mechanical defect in the joint, whereas false ankylosis is a condition in which there is a restriction in mandibular movement due to muscular spasm myositis ossificans, and coronoid process hyperplasia. Ankylosis is characterized by the inability to open the mouth due to fusion of the TMJ condyle to the base of the skull as a result of trauma, infection, or systemic diseases such as rheumatoid arthritis (the most common) and psoraisis. Ankylosis causes facial asymmetry and affects the patient psychologically as well as speech, difficult mastication, poor oral hygiene, malocclusion, and other factors. TMJ is a technically challenging joint; hence TMJ ankylosis management is complicated. Case presentation: this case is a male patient 25 years old reported to our maxillofacial clinic in Damietta faculty of medicine, Al-Azhar University with the inability to open the mouth at all, with a history of difficulty of mouth breathing and eating foods, there was a history of falling from height at 2006, and the patient underwent corrective surgery before with no improvement because the ankylosis was relapsed short period after the previous operations with that done out of our hospital inter-incisor distant ZERO so, this condition need mandatory management. Clinical examination and radiological investigations were done after complete approval from the patient and his brother; tracheostomy was done for our patient before the operation. The patient entered the operation in our hospital and drastic improvement in mouth opening was noticed, helping to restore the physical psychological health of the patient.

Keywords: temporomandibular joint, TMJ, Ankylosis, mouth opening, physiotherapy, condylar plate

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Authors: Michael A. Sprayberry, Vincent C. Paquit

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Process parameter optimization in metal powder bed electron beam melting (MPBEBM) is crucial to ensure the technology's repeatability, control, and industry-continued adoption. Despite continued efforts to address the challenges via the traditional design of experiments and process mapping techniques, there needs to be more successful in an on-the-fly optimization framework that can be adapted to MPBEBM systems. Additionally, data-intensive physics-based modeling and simulation methods are difficult to support by a metal AM alloy or system due to cost restrictions. To mitigate the challenge of resource-intensive experiments and models, this paper introduces a Multi-Objective Reinforcement Learning (MORL) methodology defined as an optimization problem for MPBEBM. An off-policy MORL framework based on policy gradient is proposed to discover optimal sets of beam power (P) – beam velocity (v) combinations to maintain a steady-state melt pool depth and phase transformation. For this, an experimentally validated Eagar-Tsai melt pool model is used to simulate the MPBEBM environment, where the beam acts as the agent across the P – v space to maximize returns for the uncertain powder bed environment producing a melt pool and phase transformation closer to the optimum. The culmination of the training process yields a set of process parameters {power, speed, hatch spacing, layer depth, and preheat} where the state (P,v) with the highest returns corresponds to a refined process parameter mapping. The resultant objects and mapping of returns to the P-v space show convergence with experimental observations. The framework, therefore, provides a model-free multi-objective approach to discovery without the need for trial-and-error experiments.

Keywords: additive manufacturing, metal powder bed fusion, reinforcement learning, process parameter optimization

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Authors: Michelangelo Sofo, Giuseppe Labianca

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In recent years, the scientific community's interest in the exploratory analysis of biomedical data has increased exponentially. Considering the field of research of nutritional biologists, the curative process, based on the analysis of clinical data, is a very delicate operation due to the fact that there are multiple solutions for the management of pathologies in the food sector (for example can recall intolerances and allergies, management of cholesterol metabolism, diabetic pathologies, arterial hypertension, up to obesity and breathing and sleep problems). In this regard, in this research work a system was created capable of evaluating various dietary regimes for specific patient pathologies. The system is founded on a mathematical-numerical model and has been created tailored for the real working needs of an expert in human nutrition using the human-centered design (ISO 9241-210), therefore it is in step with continuous scientific progress in the field and evolves through the experience of managed clinical cases (machine learning process). DietAdhoc® is a decision support system nutrition specialists for patients of both sexes (from 18 years of age) developed with an agile methodology. Its task consists in drawing up the biomedical and clinical profile of the specific patient by applying two algorithmic optimization approaches on nutritional data and a symbolic solution, obtained by transforming the relational database underlying the system into a deductive database. For all three solution approaches, particular emphasis has been given to the explainability of the suggested clinical decisions through flexible and customizable user interfaces. Furthermore, the system has multiple software modules based on time series and visual analytics techniques that allow to evaluate the complete picture of the situation and the evolution of the diet assigned for specific pathologies.

Keywords: medical decision support, physiological data extraction, data driven diagnosis, human centered AI, symbiotic AI paradigm

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Authors: Yunbin Deng

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Most existing applications of automatic speech recognition relies on cooperative subjects at a short distance to a microphone. Standoff speech recognition using microphone arrays can extend the subject to sensor distance somewhat, but it is still limited to only a few feet. As such, most deployed applications of standoff speech recognitions are limited to indoor use at short range. Moreover, these applications require air passway between the subject and the sensor to achieve reasonable signal to noise ratio. This study reports long range (50 feet) automatic speech recognition experiments using a Laser Doppler Vibrometer (LDV) sensor. This study shows that the LDV sensor modality can extend the speech acquisition standoff distance far beyond microphone arrays to hundreds of feet. In addition, LDV enables 'listening' through the windows for uncooperative subjects. This enables new capabilities in automatic audio and speech intelligence, surveillance, and reconnaissance (ISR) for law enforcement, homeland security and counter terrorism applications. The Polytec LDV model OFV-505 is used in this study. To investigate the impact of different vibrating materials, five parallel LDV speech corpora, each consisting of 630 speakers, are collected from the vibrations of a glass window, a metal plate, a plastic box, a wood slate, and a concrete wall. These are the common materials the application could encounter in a daily life. These data were compared with the microphone counterpart to manifest the impact of various materials on the spectrum of the LDV speech signal. State of the art deep neural network modeling approaches is used to conduct continuous speaker independent speech recognition on these LDV speech datasets. Preliminary phoneme recognition results using time-delay neural network, bi-directional long short term memory, and model fusion shows great promise of using LDV for long range speech recognition. To author’s best knowledge, this is the first time an LDV is reported for long distance speech recognition application.

Keywords: covert speech acquisition, distant speech recognition, DSR, laser Doppler vibrometer, LDV, speech intelligence surveillance and reconnaissance, ISR

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Authors: Dan Frederick Orcherton

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This research explores the fusion of "Critical Pedagogy of Place" and "Indigenous Education Philosophy" to create a holistic pedagogical framework within Instructional Theory, focusing on its application in Distance Education, specifically within two Pacific Island cultures. The study's objectives included investigating culturally relevant instructional techniques, strategies, and technologies for the Itaukei (Indigenous Fijian) and ni-Vanuatu cultures, enhancing appreciation for culturally sensitive pedagogical methods. Methodologically, a qualitative inquiry phenomenological approach was employed within a constructivist paradigm, utilizing a comprehensive qualitative scoping review and online literature search. Key findings include the prioritization of cultural inclusivity and indigenous knowledge integration in both indigenous education philosophies and various instructional approaches. Learner-centered methods like constructivist andragogy and the learning cycle are applicable and effective in distance education within these cultures, aligning with indigenous learners' values and preferences. Place-based education and critical pedagogy of place are particularly pertinent, fostering a deeper connection between education, local environments, and social justice. Integrating digital technologies in culturally responsive education bridges geographical gaps and preserves cultural knowledge. Lastly, blending Western and Indigenous Science, influenced by a Two-Eyed Seeing approach, informs pedagogy by combining Western and Indigenous Science. This research underscores the importance of acknowledging cultural diversity and respecting indigenous knowledge in distance education. It highlights the value of learner-centered approaches, place-based education, and technology integration. The study enriches the educational experience within the Itaukei and niVanuatu cultures and provides insights for educators and policymakers aiming to bridge cultural gaps in distance education.

Keywords: critical pedegory of place, Itaukei (indigenous Fijian) and ni-Vanuatu cultures, placed-based education, indigenous knowledge, distance education

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Authors: Farha Mizana Shamsudin, Shahidan Radiman, Yusof Abdullah, Nasri Abdul Hamid

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Oxide dispersion strengthened (ODS) ferritic steels are amongst the most promising candidates for large scale structural materials to be applied in next generation fission and fusion nuclear power reactors. This kind of material is relatively stable at high temperature, possess remarkable mechanical properties and comparatively good resistance from neutron radiation damage. The superior performance of ODS ferritic steels over their conventional properties is attributed to the high number density of nano-sized dispersoids that act as nucleation sites and stable sinks for many small helium bubbles resulting from irradiation, and also as pinning points to dislocation movement and grain growth. ODS ferritic steels are usually produced by powder metallurgical routes involving mechanical alloying (MA) process of Y2O3 and pre-alloyed or elemental metallic powders, and then consolidated by hot isostatic pressing (HIP) or hot extrusion (HE) techniques. In this study, Fe-14Cr-3W-0.5Ti-0.3Y₂O₃ (designated as 14YWT) was produced by mechanical alloying process and followed by hot isostatic pressing (HIP) technique. Crystal structure and morphology of this sample were identified and characterized by using X-ray Diffraction (XRD) and field emission scanning electron microscope (FESEM) respectively. The magnetic measurement of this sample at room temperature was carried out by using a vibrating sample magnetometer (VSM). FESEM micrograph revealed a homogeneous microstructure constituted by fine grains of less than 650 nm in size. The ultra-fine dispersoids of size between 5 nm to 19 nm were observed homogeneously distributed within the BCC matrix. The EDS mapping reveals that the dispersoids contain Y-Ti-O nanoclusters and from the magnetization curve plotted by VSM, this sample approaches the behavior of soft ferromagnetic materials. In conclusion, ODS Fe-14Cr-3W-0.5Ti-0.3Y₂O₃ (14YWT) ferritic steel was successfully produced by HIP technique in this present study.

Keywords: hot isostatic pressing, magnetization, microstructure, ODS ferritic steel

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Authors: Billy Moore, Izaiah Smith, Dominic Mckinney, Andrew Cisco, Mehdi Kabir

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Phase-change materials (PCMs) are considered one of the most promising substances to be engaged passively in thermal management and storage systems for spacecraft, where it is critical to diminish the overall mass of the onboard thermal storage system while minimizing temperature fluctuations upon drastic changes in the environmental temperature within the orbit stage. This makes the development of effective thermal management systems more challenging since there is no atmosphere in outer space to take advantage of natural and forced convective heat transfer. PCM can store or release a tremendous amount of thermal energy within a small volume in the form of latent heat of fusion in the phase-change processes of melting and solidification from solid to liquid or, conversely, during which temperature remains almost constant. However, the existing PCMs pose very low thermal conductivity, leading to an undesirable increase in total thermal resistance and, consequently, a slow thermal response time. This often turns into a system bottleneck from the thermal performance perspective. To address the above-mentioned drawback, the present study aims to design and develop various heat sinks featured by nano-structured graphitic foams (i.e., carbon foam), expanded graphite (EG), and open-cell copper foam (OCCF) infiltrated with a conventional paraffin wax PCM with a melting temperature of around 35 °C. This study focuses on the use of passive thermal management techniques to develop efficient heat sinks to maintain the electronics circuits’ and battery module’s temperature within the thermal safety limit for small spacecraft and satellites such as the Pumpkin and OPTIMUS battery modules designed for CubeSats with a cross-sectional area of approximately 4˝×4˝. Thermal response times for various heat sinks are assessed in a vacuum chamber to simulate space conditions.

Keywords: heat sink, porous foams, phase-change material (PCM), spacecraft thermal management

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Authors: Muhammad Awais Afzal, Lorena Tuchscherr De Hauschopp, Christian Hübner

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Introduction: Autophagy is an evolutionarily conserved lysosome-dependent degradation pathway, which can be induced by extrinsic and intrinsic stressors in living systems to adapt to fluctuating environmental conditions. In the context of inflammatory stress, autophagy contributes to the elimination of invading pathogens, the regulation of innate and adaptive immune mechanisms, and regulation of inflammasome activity as well as tissue damage repair. Lysosomes can be recycled from autolysosomes by the process of autophagic lysosome reformation (ALR), which depends on the presence of several proteins including Spatacsin. Thus ALR contributes to the replenishment of lysosomes that are available for fusion with autophagosomes in situations of increased autophagic turnover, e.g., during bacterial infections, inflammatory stress or sepsis. Objectives: We aimed to assess whether ALR plays a role for cell survival in an in-vitro bacterial infection model. Methods: Mouse embryonic fibroblasts (MEFs) were isolated from wild-type mice and Spatacsin (Spg11-/-) knockout mice. Wild-type MEFs and Spg11-/- MEFs were infected with Staphylococcus aureus (multiplication of infection (MOI) used was 10). After 8 and 16 hours of infection, cell viability was assessed on BD flow cytometer through propidium iodide intake. Bacterial intake by cells was also calculated by plating cell lysates on blood agar plates. Results: in-vitro infection of MEFs with Staphylococcus aureus showed a marked decrease of cell viability in ALR deficient Spatacsin knockout (Spg11-/-) MEFs after 16 hours of infection as compared to wild-type MEFs (n=3 independent experiments; p < 0.0001) although no difference was observed for bacterial intake by both genotypes. Conclusion: Suggesting that ALR is important for the defense of invading pathogens e.g. S. aureus, we observed a marked increase of cell death in an in-vitro infection model in cells with compromised ALR.

Keywords: autophagy, autophagic lysosome reformation, bacterial infections, Staphylococcus aureus

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