Search results for: isolated word recognition
Commenced in January 2007
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Edition: International
Paper Count: 3865

Search results for: isolated word recognition

265 Globalisation and Diplomacy: How Can Small States Improve the Practice of Diplomacy to Secure Their Foreign Policy Objectives?

Authors: H. M. Ross-McAlpine

Abstract:

Much of what is written on diplomacy, globalization and the global economy addresses the changing nature of relationships between major powers. While the most dramatic and influential changes have resulted from these developing relationships the world is not, on deeper inspection, governed neatly by major powers. Due to advances in technology, the shifting balance of power and a changing geopolitical order, small states have the ability to exercise a greater influence than ever before. Increasingly interdependent and ever complex, our world is too delicate to be handled by a mighty few. The pressure of global change requires small states to adapt their diplomatic practices and diversify their strategic alliances and relationships. The nature and practice of diplomacy must be re-evaluated in light of the pressures resulting from globalization. This research examines: how small states can best secure their foreign policy objectives? Small state theory is used as a foundation for exploring the case study of New Zealand. The research draws on secondary sources to evaluate the existing theory in relation to modern practices of diplomacy. As New Zealand lacks the required economic and military power to play an active, influential role in international affairs what strategies are used to exert influence? Furthermore, New Zealand lies in a remote corner of the Pacific and is geographically isolated from its nearest neighbors how does this affect security and trade priorities? The findings note a significant shift since the 1970’s in New Zealand’s diplomatic relations. This shift is arguably a direct result of globalization, regionalism and a growing independence from the traditional bi-lateral relationships. The need to source predictable trade, investment and technology are an essential driving force for New Zealand’s diplomatic relations. A lack of hard power aligns New Zealand’s prosperity with a secure, rules-based international system that increases the likelihood of a stable and secure global order. New Zealand’s diplomacy and prosperity has been intrinsically reliant on its reputation. A vital component of New Zealand’s diplomacy is preserving a reputation for integrity and global responsibility. It is the use of this soft power that facilitates the influence that New Zealand enjoys on the world stage. To weave a comprehensive network of successful diplomatic relationships, New Zealand must maintain a reputation of international credibility. Globalization has substantially influenced the practice of diplomacy for New Zealand. The current world order places economic and military might in the hands of a few, subsequently requiring smaller states to use other means for securing their interests. There are clear strategies evident in New Zealand’s diplomacy practice that draw attention to how other smaller states might best secure their foreign policy objectives. While these findings are limited, as with all case study research, there is value in applying the findings to other small states struggling to secure their interests in the wake of rapid globalization.

Keywords: diplomacy, foreign policy, globalisation, small state

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264 Development of a Miniature Laboratory Lactic Goat Cheese Model to Study the Expression of Spoilage by Pseudomonas Spp. In Cheeses

Authors: Abirami Baleswaran, Christel Couderc, Loubnah Belahcen, Jean Dayde, Hélène Tormo, Gwénaëlle Jard

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Cheeses are often reported to be spoiled by Pseudomonas spp., responsible for defects in appearance, texture, taste, and smell, leading to their non-marketing and even their destruction. Despite preventive actions, problems linked to Pseudomonas spp. are difficult to control by the lack of knowledge and control of these contaminants during the cheese manufacturing. Lactic goat cheese producers are not spared by this problem and are looking for solutions to decrease the number of spoiled cheeses. To explore different hypotheses, experiments are needed. However, cheese-making experiments at the pilot scale are expensive and time consuming. Thus, there is a real need to develop a miniature cheeses model system under controlled conditions. In a previous study, several miniature cheese models corresponding to different type of commercial cheeses have been developed for different purposes. The models were, for example, used to study the influence of milk, starters cultures, pathogen inhibiting additives, enzymatic reactions, microflora, freezing process on cheese. Nevertheless, no miniature model was described on the lactic goat cheese. The aim of this work was to develop a miniature cheese model system under controlled laboratory conditions which resembles commercial lactic goat cheese to study Pseudomonas spp. spoilage during the manufacturing and ripening process. First, a protocol for the preparation of miniature cheeses (3.5 times smaller than a commercial one) was designed based on the cheese factorymanufacturing process. The process was adapted from “Rocamadour” technology and involves maturation of pasteurized milk, coagulation, removal of whey by centrifugation, moulding, and ripening in a little scale cellar. Microbiological (total bacterial count, yeast, molds) and physicochemical (pH, saltinmoisture, moisture in fat-free)analyses were performed on four key stages of the process (before salting, after salting, 1st day of ripening, and end of ripening). Factory and miniature cheeses volatilomewere also obtained after full scan Sift-MS cheese analysis. Then, Pseudomonas spp. strains isolated from contaminated cheeses were selected on their origin, their ability to produce pigments, and their enzymatic activities (proteolytic, lecithinasic, and lipolytic). Factory and miniature curds were inoculated by spotting selected strains on the cheese surface. The expression of cheese spoilage was evaluated by counting the level of Pseudomonas spp. during the ripening and by visual observation and under UVlamp. The physicochemical and microbiological compositions of miniature cheeses permitted to assess that miniature process resembles factory process. As expected, differences involatilomes were observed, probably due to the fact that miniature cheeses are made usingpasteurized milk to better control the microbiological conditions and also because the little format of cheese induced probably a difference during the ripening even if the humidity and temperature in the cellar were quite similar. The spoilage expression of Pseudomonas spp. was observed in miniature and factory cheeses. It confirms that the proposed model is suitable for the preparation of miniature cheese specimens in the spoilage study of Pseudomonas spp. in lactic cheeses. This kind of model could be deployed for other applications and other type of cheese.

Keywords: cheese, miniature, model, pseudomonas spp, spoilage

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263 Characterization and Evaluation of the Dissolution Increase of Molecular Solid Dispersions of Efavirenz

Authors: Leslie Raphael de M. Ferraz, Salvana Priscylla M. Costa, Tarcyla de A. Gomes, Giovanna Christinne R. M. Schver, Cristóvão R. da Silva, Magaly Andreza M. de Lyra, Danilo Augusto F. Fontes, Larissa A. Rolim, Amanda Carla Q. M. Vieira, Miracy M. de Albuquerque, Pedro J. Rolim-Neto

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Efavirenz (EFV) is a drug used as first-line treatment of AIDS. However, it has poor aqueous solubility and wettability, presenting problems in the gastrointestinal tract absorption and bioavailability. One of the most promising strategies to improve the solubility is the use of solid dispersions (SD). Therefore, this study aimed to characterize SD EFZ with the polymers: PVP-K30, PVPVA 64 and SOLUPLUS in order to find an optimal formulation to compose a future pharmaceutical product for AIDS therapy. Initially, Physical Mixtures (PM) and SD with the polymers were obtained containing 10, 20, 50 and 80% of drug (w/w) by the solvent method. The best formulation obtained between the SD was selected by in vitro dissolution test. Finally, the drug-carrier system chosen, in all ratios obtained, were analyzed by the following techniques: Differential Scanning Calorimetry (DSC), polarization microscopy, Scanning Electron Microscopy (SEM) and spectrophotometry of absorption in the region of infrared (IR). From the dissolution profiles of EFV, PM and SD, the values of area Under The Curve (AUC) were calculated. The data showed that the AUC of all PM is greater than the isolated EFV, this result is derived from the hydrophilic properties of the polymers thus favoring a decrease in surface tension between the drug and the dissolution medium. In adittion, this ensures an increasing of wettability of the drug. In parallel, it was found that SD whom had higher AUC values, were those who have the greatest amount of polymer (with only 10% drug). As the amount of drug increases, it was noticed that these results either decrease or are statistically similar. The AUC values of the SD using the three different polymers, followed this decreasing order: SD PVPVA 64-EFV 10% > SD PVP-K30-EFV 10% > SD Soluplus®-EFV 10%. The DSC curves of SD’s did not show the characteristic endothermic event of drug melt process, suggesting that the EFV was converted to its amorphous state. The analysis of polarized light microscopy showed significant birefringence of the PM’s, but this was not observed in films of SD’s, thus suggesting the conversion of the drug from the crystalline to the amorphous state. In electron micrographs of all PM, independently of the percentage of the drug, the crystal structure of EFV was clearly detectable. Moreover, electron micrographs of the SD with the two polymers in different ratios investigated, we observed the presence of particles with irregular size and morphology, also occurring an extensive change in the appearance of the polymer, not being possible to differentiate the two components. IR spectra of PM corresponds to the overlapping of polymer and EFV bands indicating thereby that there is no interaction between them, unlike the spectra of all SD that showed complete disappearance of the band related to the axial deformation of the NH group of EFV. Therefore, this study was able to obtain a suitable formulation to overcome the solubility limitations of the EFV, since SD PVPVA 64-EFZ 10% was chosen as the best system in delay crystallization of the prototype, reaching higher levels of super saturation.

Keywords: characterization, dissolution, Efavirenz, solid dispersions

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262 Microbiological and Physicochemical Evaluation of Traditional Greek Kopanisti Cheese Produced by Different Starter Cultures

Authors: M. Kazou, A. Gavriil, O. Kalagkatsi, T. Paschos, E. Tsakalidou

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Kopanisti cheese is a Greek soft Protected Designation of Origin (PDO) cheese made of raw cow, sheep or goat milk, or mixtures of them, with similar organoleptic characteristics to that of Roquefort cheese. Traditional manufacturing of Kopanisti cheese is limited in small-scale dairies, without the addition of starter cultures. Instead, an amount of over-mature Kopanisti cheese, called Mana Kopanisti, is used to initiate ripening. Therefore, the selection of proper starter cultures and the understanding of the contribution of various microbial groups to its overall quality is crucial for the production of a high-quality final product with standardized organoleptic and physicochemical characteristics. Taking the above into account, the aim of the present study was the investigation of Kopanisti cheese microbiota and its role in cheese quality. For this purpose, four different types of Kopanisti were produced in triplicates, all with pasteurized cow milk, with the addition of (A) the typical mesophilic species Lactococcus lactis and Lactobacillus paracasei used as starters in the production of soft spread cheeses, (B) strains of Lactobacillus acidipiscis and Lactobacillus rennini previously isolated from Kopanisti and Mana Kopanisti, (C) all the species from (A) and (B) as inoculum, and finally (D) the species from (A) and Mana Kopanisti. Physicochemical and microbiological analysis was performed for milk and cheese samples during ripening. Enumeration was performed for major groups of lactic acid bacteria (LAB), total mesophilic bacteria, yeasts as well as hygiene indicator microorganisms. Bacterial isolates from all the different LAB groups, apart from enterococci, alongside yeasts isolates, were initially grouped using repetitive sequence-based polymerase chain reaction (rep-PCR) and then identified at the species level using 16S rRNA gene and internal transcribed spacer (ITS) DNA region sequencing, respectively. Sensory evaluation was also performed for final cheese samples at the end of the ripening period (35 days). Based on the results of the classical microbiological analysis, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, ranged between 7 and 10 log colony forming unit (CFU) g⁻¹, phychrotrophic bacteria, and yeast extract glucose chloramphenicol (YGC) isolates between 4 and 8 log CFU g⁻¹, while coliforms and enterococci up to 2 log CFU g⁻¹ throughout ripening in cheese samples A, C and D. In contrast, in cheese sample B, the average counts of the total mesophilic bacteria and LAB, apart from enterococci, phychrotrophic bacteria, and YGC isolates ranged between 0 and 10 log CFU g⁻¹ and coliforms and enterococci up to 2 log CFU g⁻¹. Although the microbial counts were not that different among samples, identification of the bacterial and yeasts isolates revealed the complex microbial community structure present in each cheese sample. Differences in the physicochemical characteristics among the cheese samples were also observed, with pH ranging from 4.3 to 5.3 and moisture from 49.6 to 58.0 % in the final cheese products. Interestingly, the sensory evaluation also revealed differences among samples, with cheese sample B ranking first based on the total score. Overall, the combination of these analyses highlighted the impact of different starter cultures on the Kopanisti microbiota as well as on the physicochemical and sensory characteristics of the final product.

Keywords: Kopanisti cheese, microbiota, classical microbiological analysis, physicochemical analysis

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261 Gold Nanoprobes Assay for the Identification of Foodborn Pathogens Such as Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis

Authors: D. P. Houhoula, J. Papaparaskevas, S. Konteles, A. Dargenta, A. Farka, C. Spyrou, M. Ziaka, S. Koussisis, E. Charvalos

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Objectives: Nanotechnology is providing revolutionary opportunities for the rapid and simple diagnosis of many infectious diseases. Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritis are important human pathogens. Diagnostic assays for bacterial culture and identification are time consuming and laborious. There is an urgent need to develop rapid, sensitive, and inexpensive diagnostic tests. In this study, a gold nanoprobe strategy developed and relies on the colorimetric differentiation of specific DNA sequences based approach on differential aggregation profiles in the presence or absence of specific target hybridization. Method: Gold nanoparticles (AuNPs) were purchased from Nanopartz. They were conjugated with thiolated oligonucleotides specific for the femA gene for the identification of members of Staphylococcus aureus, the mecA gene for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus, hly gene encoding the pore-forming cytolysin listeriolysin for the identification of Listeria monocytogenes and the invA sequence for the identification of Salmonella enteritis. DNA isolation from Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis cultures was performed using the commercial kit Nucleospin Tissue (Macherey Nagel). Specifically 20μl of DNA was diluted in 10mMPBS (pH5). After the denaturation of 10min, 20μl of AuNPs was added followed by the annealing step at 58oC. The presence of a complementary target prevents aggregation with the addition of acid and the solution remains pink, whereas in the opposite event it turns to purple. The color could be detected visually and it was confirmed with an absorption spectrum. Results: Specifically, 0.123 μg/μl DNA of St. aureus, L.monocytogenes and Salmonella enteritis was serially diluted from 1:10 to 1:100. Blanks containing PBS buffer instead of DNA were used. The application of the proposed method on isolated bacteria produced positive results with all the species of St. aureus and L. monocytogenes and Salmonella enteritis using the femA, mecA, hly and invA genes respectively. The minimum detection limit of the assay was defined at 0.2 ng/μL of DNA. Below of 0.2 ng/μL of bacterial DNA the solution turned purple after addition of HCl, defining the minimum detection limit of the assay. None of the blank samples was positive. The specificity was 100%. The application of the proposed method produced exactly the same results every time (n = 4) the evaluation was repeated (100% repeatability) using the femA, hly and invA genes. Using the gene mecA for the differentiation of Staphylococcus aureus and MRSA Staphylococcus aureus the method had a repeatability 50%. Conclusion: The proposed method could be used as a highly specific and sensitive screening tool for the detection and differentiation of Staphylococcus aureus Listeria monocytogenes and Salmonella enteritis. The use AuNPs for the colorimetric detection of DNA targets represents an inexpensive and easy-to-perform alternative to common molecular assays. The technology described here, may develop into a platform that could accommodate detection of many bacterial species.

Keywords: gold nanoparticles, pathogens, nanotechnology, bacteria

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260 Multi-Labeled Aromatic Medicinal Plant Image Classification Using Deep Learning

Authors: Tsega Asresa, Getahun Tigistu, Melaku Bayih

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Computer vision is a subfield of artificial intelligence that allows computers and systems to extract meaning from digital images and video. It is used in a wide range of fields of study, including self-driving cars, video surveillance, medical diagnosis, manufacturing, law, agriculture, quality control, health care, facial recognition, and military applications. Aromatic medicinal plants are botanical raw materials used in cosmetics, medicines, health foods, essential oils, decoration, cleaning, and other natural health products for therapeutic and Aromatic culinary purposes. These plants and their products not only serve as a valuable source of income for farmers and entrepreneurs but also going to export for valuable foreign currency exchange. In Ethiopia, there is a lack of technologies for the classification and identification of Aromatic medicinal plant parts and disease type cured by aromatic medicinal plants. Farmers, industry personnel, academicians, and pharmacists find it difficult to identify plant parts and disease types cured by plants before ingredient extraction in the laboratory. Manual plant identification is a time-consuming, labor-intensive, and lengthy process. To alleviate these challenges, few studies have been conducted in the area to address these issues. One way to overcome these problems is to develop a deep learning model for efficient identification of Aromatic medicinal plant parts with their corresponding disease type. The objective of the proposed study is to identify the aromatic medicinal plant parts and their disease type classification using computer vision technology. Therefore, this research initiated a model for the classification of aromatic medicinal plant parts and their disease type by exploring computer vision technology. Morphological characteristics are still the most important tools for the identification of plants. Leaves are the most widely used parts of plants besides roots, flowers, fruits, and latex. For this study, the researcher used RGB leaf images with a size of 128x128 x3. In this study, the researchers trained five cutting-edge models: convolutional neural network, Inception V3, Residual Neural Network, Mobile Network, and Visual Geometry Group. Those models were chosen after a comprehensive review of the best-performing models. The 80/20 percentage split is used to evaluate the model, and classification metrics are used to compare models. The pre-trained Inception V3 model outperforms well, with training and validation accuracy of 99.8% and 98.7%, respectively.

Keywords: aromatic medicinal plant, computer vision, convolutional neural network, deep learning, plant classification, residual neural network

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259 Neuroprotection against N-Methyl-D-Aspartate-Induced Optic Nerve and Retinal Degeneration Changes by Philanthotoxin-343 to Alleviate Visual Impairments Involve Reduced Nitrosative Stress

Authors: Izuddin Fahmy Abu, Mohamad Haiqal Nizar Mohamad, Muhammad Fattah Fazel, Renu Agarwal, Igor Iezhitsa, Nor Salmah Bakar, Henrik Franzyk, Ian Mellor

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Glaucoma is the global leading cause of irreversible blindness. Currently, the available treatment strategy only involves lowering intraocular pressure (IOP); however, the condition often progresses despite lowered or normal IOP in some patients. N-methyl-D-aspartate receptor (NMDAR) excitotoxicity often occurs in neurodegeneration-related glaucoma; thus it is a relevant target to develop a therapy based on neuroprotection approach. This study investigated the effects of Philanthotoxin-343 (PhTX-343), an NMDAR antagonist, on the neuroprotection of NMDA-induced glaucoma to alleviate visual impairments. Male Sprague-Dawley rats were equally divided: Groups 1 (control) and 2 (glaucoma) were intravitreally injected with phosphate buffer saline (PBS) and NMDA (160nM), respectively, while group 3 was pre-treated with PhTX-343 (160nM) 24 hours prior to NMDA injection. Seven days post-treatments, rats were subjected to visual behavior assessments and subsequently euthanized to harvest their retina and optic nerve tissues for histological analysis and determination of nitrosative stress level using 3-nitrotyrosine ELISA. Visual behavior assessments via open field, object, and color recognition tests demonstrated poor visual performance in glaucoma rats indicated by high exploratory behavior. PhTX-343 pre-treatment appeared to preserve visual abilities as all test results were significantly improved (p < 0.05). H&E staining of the retina showed a marked reduction of ganglion cell layer thickness in the glaucoma group; in contrast, PhTX-343 significantly increased the number by 1.28-folds (p < 0.05). PhTX-343 also increased the number of cell nuclei/100μm2 within inner retina by 1.82-folds compared to the glaucoma group (p < 0.05). Toluidine blue staining of optic nerve tissues showed that PhTX-343 reduced the degeneration changes compared to the glaucoma group which exhibited vacuolation overall sections. PhTX-343 also decreased retinal 3- nitrotyrosine concentration by 1.74-folds compared to the glaucoma group (p < 0.05). All results in PhTX-343 group were comparable to control (p > 0.05). We conclude that PhTX-343 protects against NMDA-induced changes and visual impairments in the rat model by reducing nitrosative stress levels.

Keywords: excitotoxicity, glaucoma, nitrosative stress , NMDA receptor , N-methyl-D-aspartate , philanthotoxin, visual behaviour

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258 A Player's Perspective of University Elite Netball Programmes in South Africa

Authors: Wim Hollander, Petrus Louis Nolte

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University sport in South Africa is not isolated from the complexity of globalization and professionalization of sport, as it forms an integral part of the sports development environment in South Africa. In order to align their sports programs with global and professional requirements, several universities opted to develop elite sports programs; recruit specialized personnel such as coaches, administrators, and athletes; provide expert coaching; scientific and medical services; sports testing; fitness, technical and tactical expertise; sport psychological and rehabilitation support; academic guidance and career assistance; and student-athlete accommodation. In addition, universities provide administrative support and high-quality physical resources (training facilities) for the benefit of the overall South African sport system. Although it is not compulsory for universities to develop elite sports programs to prepare their teams for competitions, elite competitions such as the annual Varsity Sport, University Sport South Africa (USSA) and local club competitions and leagues within international university competitions where universities not only compete but also deliver players for representative national netball teams. The aim of this study is, therefore, to describe the perceptions of players of the university elite netball programs they were participating in. This study adopted a descriptive design with a quantitative approach, utilizing a self-structured questionnaire as a research technique. As this research formed part of a national research project for NSA with a population of 172 national and provincial netball players, a sample of 92 university netball players from the population was selected. Content validity of the self-structured questionnaire was secured through a test-retest process, with construct validity through a member of the Statistical Consultation Services (STATCON) of the University of Johannesburg that provided feedback on the structural format of the questionnaire. Reliability was measured utilizing Cronbach Alpha on p < 0.005 level of significance. A reliability score of 0.87 was measured. The research was approved by the Board of Netball South Africa and ethical conduct implemented according to the processes and procedures approved by the Ethics Committees of the Faculty of Health Sciences, the University of Johannesburg with clearance number REC-01-30-2019. From the results, it is evident that university elite netball programs are professional, especially with regards to the employment of knowledgeable and competent coaches and technical officials such as team managers and sport sciences staff. These professionals have access to elite training facilities, support staff, and relatively large groups of elite players, all elements of an elite program that could enhance the national federation’s (Netball South Africa) system. Universities could serve the dual purpose of serving as university netball clubs, as well as providing elite training services and facilities as performance hubs for national players.

Keywords: elite sport programmes, university netball, player experiences, varsity sport netball

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257 Video Analytics on Pedagogy Using Big Data

Authors: Jamuna Loganath

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Education is the key to the development of any individual’s personality. Today’s students will be tomorrow’s citizens of the global society. The education of the student is the edifice on which his/her future will be built. Schools therefore should provide an all-round development of students so as to foster a healthy society. The behaviors and the attitude of the students in school play an essential role for the success of the education process. Frequent reports of misbehaviors such as clowning, harassing classmates, verbal insults are becoming common in schools today. If this issue is left unattended, it may develop a negative attitude and increase the delinquent behavior. So, the need of the hour is to find a solution to this problem. To solve this issue, it is important to monitor the students’ behaviors in school and give necessary feedback and mentor them to develop a positive attitude and help them to become a successful grownup. Nevertheless, measuring students’ behavior and attitude is extremely challenging. None of the present technology has proven to be effective in this measurement process because actions, reactions, interactions, response of the students are rarely used in the course of the data due to complexity. The purpose of this proposal is to recommend an effective supervising system after carrying out a feasibility study by measuring the behavior of the Students. This can be achieved by equipping schools with CCTV cameras. These CCTV cameras installed in various schools of the world capture the facial expressions and interactions of the students inside and outside their classroom. The real time raw videos captured from the CCTV can be uploaded to the cloud with the help of a network. The video feeds get scooped into various nodes in the same rack or on the different racks in the same cluster in Hadoop HDFS. The video feeds are converted into small frames and analyzed using various Pattern recognition algorithms and MapReduce algorithm. Then, the video frames are compared with the bench marking database (good behavior). When misbehavior is detected, an alert message can be sent to the counseling department which helps them in mentoring the students. This will help in improving the effectiveness of the education process. As Video feeds come from multiple geographical areas (schools from different parts of the world), BIG DATA helps in real time analysis as it analyzes computationally to reveal patterns, trends, and associations, especially relating to human behavior and interactions. It also analyzes data that can’t be analyzed by traditional software applications such as RDBMS, OODBMS. It has also proven successful in handling human reactions with ease. Therefore, BIG DATA could certainly play a vital role in handling this issue. Thus, effectiveness of the education process can be enhanced with the help of video analytics using the latest BIG DATA technology.

Keywords: big data, cloud, CCTV, education process

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256 Analyzing the Use of Augmented and Virtual Reality to Teach Social Skills to Students with Autism

Authors: Maggie Mosher, Adam Carreon, Sean Smith

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A systematic literature review was conducted to explore the evidence base on the use of augmented reality (AR), virtual reality (VR), mixed reality (MR), and extended reality (XR) to present social skill instruction to school-age students with autism spectrum disorder (ASD). Specifically, the systematic review focus was on a. the participants and intervention agents using AR, VR, MR, and XR for social skill acquisition b. the social skills taught through these mediums and c. the social validity measures (i.e., goals, procedures, and outcomes) reported in these studies. Forty-one articles met the inclusion criteria. Researchers in six studies taught social skills to students through AR, in 27 studies through non-immersive VR, and in 10 studies through immersive VR. No studies used MR or XR. The primary targeted social skills were relationship skills, emotion recognition, social awareness, cooperation, and executive functioning. An intervention to improve many social skills was implemented by 73% of researchers, 17% taught a single skill, and 10% did not clearly state the targeted skill. The intervention was considered effective in 26 of the 41 studies (63%), not effective in four studies (10%), and 11 studies (27%) reported mixed results. No researchers reported information for all 17 social validity indicators. The social validity indicators reported by researchers ranged from two to 14. Social validity measures on the feelings toward and use of the technology were provided in 22 studies (54%). Findings indicated both AR and VR are promising platforms for providing social skill instruction to students with ASD. Studies utilizing this technology show a number of social validity indicators. However, the limited information provided on the various interventions, participant characteristics, and validity measures, offers insufficient evidence of the impact of these technologies in teaching social skills to students with ASD. Future research should develop a protocol for training treatment agents to assess the role of different variables (i.e., whether agents are customizing content, monitoring student learning, using intervention specific vocabulary in their day to day instruction). Sustainability may be increased by providing training in the technology to both treatment agents and participants. Providing scripts of instruction occurring within the intervention would provide the needed information to determine the primary method of teaching within the intervention. These variables play a role in maintenance and generalization of the social skills. Understanding the type of feedback provided would help researchers determine if students were able to feel rewarded for progressing through the scenarios or if students require rewarding aspects within the intervention (i.e., badges, trophies). AR has the potential to generalize instruction and VR has the potential for providing a practice environment for performance deficits. Combining these two technologies into a mixed reality intervention may provide a more cohesive and effective intervention.

Keywords: autism, augmented reality, social and emotional learning, social skills, virtual reality

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255 Growth and Bone Health in Children following Liver Transplantation

Authors: Faris Alkhalil, Rana Bitar, Amer Azaz, Hisham Natour, Noora Almeraikhi, Mohamad Miqdady

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Background: Children with liver transplantation are achieving very good survival and so there is now a need to concentrate on achieving good health in these patients and preventing disease. Immunosuppressive medications have side effects that need to be monitored and if possible avoided. Glucocorticoids and calcineurin inhibitors are detrimental to bone and mineral homeostasis in addition steroids can also affect linear growth. Steroid sparing regimes in renal transplant children has shown to improve children’s height. Aim: We aim to review the growth and bone health of children post liver transplant by measuring bone mineral density (BMD) using dual energy X-ray absorptiometry (DEXA) scan and assessing if there is a clear link between poor growth and impaired bone health and use of long term steroids. Subjects and Methods: This is a single centre retrospective Cohort study, we reviewed the medical notes of children (0-16 years) who underwent a liver transplantation between November 2000 to November 2016 and currently being followed at our centre. Results: 39 patients were identified (25 males and 14 females), the median transplant age was 2 years (range 9 months - 16 years), and the median follow up was 6 years. Four patients received a combined transplant, 2 kidney and liver transplant and 2 received a liver and small bowel transplant. The indications for transplant included, Biliary Atresia (31%), Acute Liver failure (18%), Progressive Familial Intrahepatic Cholestasis (15%), transplantable metabolic disease (10%), TPN related liver disease (8%), Primary Hyperoxaluria (5%), Hepatocellular carcinoma (3%) and other causes (10%). 36 patients (95%) were on a calcineurin inhibitor (34 patients were on Tacrolimus and 2 on Cyclosporin). The other three patients were on Sirolimus. Low dose long-term steroids was used in 21% of the patients. A considerable proportion of the patients had poor growth. 15% were below the 3rd centile for weight for age and 21% were below the 3rd centile for height for age. Most of our patients with poor growth were not on long term steroids. 49% of patients had a DEXA scan post transplantation. 21% of these children had low bone mineral density, one patient had met osteoporosis criteria with a vertebral fracture. Most of our patients with impaired bone health were not on long term steroids. 20% of the patients who did not undergo a DEXA scan developed long bone fractures and 50% of them were on long term steroid use which may suggest impaired bone health in these patients. Summary and Conclusion: The incidence of impaired bone health, although studied in limited number of patients; was high. Early recognition and treatment should be instituted to avoid fractures and improve bone health. Many of the patients were below the 3rd centile for weight and height however there was no clear relationship between steroid use and impaired bone health, reduced weight and reduced linear height.

Keywords: bone, growth, pediatric, liver, transplantation

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254 Cognitive Radio in Aeronautic: Comparison of Some Spectrum Sensing Technics

Authors: Abdelkhalek Bouchikhi, Elyes Benmokhtar, Sebastien Saletzki

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The aeronautical field is experiencing issues with RF spectrum congestion due to the constant increase in the number of flights, aircrafts and telecom systems on board. In addition, these systems are bulky in size, weight and energy consumption. The cognitive radio helps particularly solving the spectrum congestion issue by its capacity to detect idle frequency channels then, allowing an opportunistic exploitation of the RF spectrum. The present work aims to propose a new use case for aeronautical spectrum sharing and to study the performances of three different detection techniques: energy detector, matched filter and cyclostationary detector within the aeronautical use case. The spectrum in the proposed cognitive radio is allocated dynamically where each cognitive radio follows a cognitive cycle. The spectrum sensing is a crucial step. The goal of the sensing is gathering data about the surrounding environment. Cognitive radio can use different sensors: antennas, cameras, accelerometer, thermometer, etc. In IEEE 802.22 standard, for example, a primary user (PU) has always the priority to communicate. When a frequency channel witch used by the primary user is idle, the secondary user (SU) is allowed to transmit in this channel. The Distance Measuring Equipment (DME) is composed of a UHF transmitter/receiver (interrogator) in the aircraft and a UHF receiver/transmitter on the ground. While the future cognitive radio will be used jointly to alleviate the spectrum congestion issue in the aeronautical field. LDACS, for example, is a good candidate; it provides two isolated data-links: ground-to-air and air-to-ground data-links. The first contribution of the present work is a strategy allowing sharing the L-band. The adopted spectrum sharing strategy is as follow: the DME will play the role of PU which is the licensed user and the LDACS1 systems will be the SUs. The SUs could use the L-band channels opportunely as long as they do not causing harmful interference signals which affect the QoS of the DME system. Although the spectrum sensing is a key step, it helps detecting holes by determining whether the primary signal is present or not in a given frequency channel. A missing detection on primary user presence creates interference between PU and SU and will affect seriously the QoS of the legacy radio. In this study, first brief definitions, concepts and the state of the art of cognitive radio will be presented. Then, a study of three communication channel detection algorithms in a cognitive radio context is carried out. The study is made from the point of view of functions, material requirements and signal detection capability in the aeronautical field. Then, we presented a modeling of the detection problem by three different methods (energy, adapted filter, and cyclostationary) as well as an algorithmic description of these detectors is done. Then, we study and compare the performance of the algorithms. Simulations were carried out using MATLAB software. We analyzed the results based on ROCs curves for SNR between -10dB and 20dB. The three detectors have been tested with a synthetics and real world signals.

Keywords: aeronautic, communication, navigation, surveillance systems, cognitive radio, spectrum sensing, software defined radio

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253 Emotion-Convolutional Neural Network for Perceiving Stress from Audio Signals: A Brain Chemistry Approach

Authors: Anup Anand Deshmukh, Catherine Soladie, Renaud Seguier

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Emotion plays a key role in many applications like healthcare, to gather patients’ emotional behavior. Unlike typical ASR (Automated Speech Recognition) problems which focus on 'what was said', it is equally important to understand 'how it was said.' There are certain emotions which are given more importance due to their effectiveness in understanding human feelings. In this paper, we propose an approach that models human stress from audio signals. The research challenge in speech emotion detection is finding the appropriate set of acoustic features corresponding to an emotion. Another difficulty lies in defining the very meaning of emotion and being able to categorize it in a precise manner. Supervised Machine Learning models, including state of the art Deep Learning classification methods, rely on the availability of clean and labelled data. One of the problems in affective computation is the limited amount of annotated data. The existing labelled emotions datasets are highly subjective to the perception of the annotator. We address the first issue of feature selection by exploiting the use of traditional MFCC (Mel-Frequency Cepstral Coefficients) features in Convolutional Neural Network. Our proposed Emo-CNN (Emotion-CNN) architecture treats speech representations in a manner similar to how CNN’s treat images in a vision problem. Our experiments show that Emo-CNN consistently and significantly outperforms the popular existing methods over multiple datasets. It achieves 90.2% categorical accuracy on the Emo-DB dataset. We claim that Emo-CNN is robust to speaker variations and environmental distortions. The proposed approach achieves 85.5% speaker-dependant categorical accuracy for SAVEE (Surrey Audio-Visual Expressed Emotion) dataset, beating the existing CNN based approach by 10.2%. To tackle the second problem of subjectivity in stress labels, we use Lovheim’s cube, which is a 3-dimensional projection of emotions. Monoamine neurotransmitters are a type of chemical messengers in the brain that transmits signals on perceiving emotions. The cube aims at explaining the relationship between these neurotransmitters and the positions of emotions in 3D space. The learnt emotion representations from the Emo-CNN are mapped to the cube using three component PCA (Principal Component Analysis) which is then used to model human stress. This proposed approach not only circumvents the need for labelled stress data but also complies with the psychological theory of emotions given by Lovheim’s cube. We believe that this work is the first step towards creating a connection between Artificial Intelligence and the chemistry of human emotions.

Keywords: deep learning, brain chemistry, emotion perception, Lovheim's cube

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252 Possibilities and Limits for the Development of Care in Primary Health Care in Brazil

Authors: Ivonete Teresinha Schulter Buss Heidemann, Michelle Kuntz Durand, Aline Megumi Arakawa-Belaunde, Sandra Mara Corrêa, Leandro Martins Costa Do Araujo, Kamila Soares Maciel

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Primary Health Care is defined as the level of a system of services that enables the achievement of answers to health needs. This level of care produces services and actions of attention to the person in the life cycle and in their health conditions or diseases. Primary Health Care refers to a conception of care model and organization of the health system that in Brazil seeks to reorganize the principles of the Unified Health System. This system is based on the principle of health as a citizen's right and duty of the State. Primary health care has family health as a priority strategy for its organization according to the precepts of the Unified Health System, structured in the logic of new sectoral practices, associating clinical work and health promotion. Thus, this study seeks to know the possibilities and limits of the care developed by professionals working in Primary Health Care. It was conducted by a qualitative approach of the participant action type, based on Paulo Freire's Research Itinerary, which corresponds to three moments: Thematic Investigation; Encoding and Decoding; and, Critical Unveiling. The themes were investigated in a health unit with the development of a culture circle with 20 professionals, from a municipality in southern Brazil, in the first half of 2021. The participants revealed as possibilities the involvement, bonding and strengthening of the interpersonal relationships of the professionals who work in the context of primary care. Promoting welcoming in primary care has favoured care and teamwork, as well as improved access. They also highlighted that care planning, the use of technologies in the process of communication and the orientation of the population enhances the levels of problem-solving capacity and the organization of services. As limits, the lack of professional recognition and the scarce material and human resources were revealed, conditions that generate tensions for health care. The reduction in the number of professionals and the low salary are pointed out as elements that boost the motivation of the health team for the development of the work. The participants revealed that due to COVID-19, the flow of care had as a priority the pandemic situation, which affected health care in primary care, and prevention and health promotion actions were canceled. The study demonstrated that empowerment and professional involvement are fundamental to promoting comprehensive and problem-solving care. However, limits of the teams are observed when exercising their activities, these are related to the lack of human and material resources, and the expansion of public health policies is urgent.

Keywords: health promotion, primary health care, health professionals, welcoming.

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251 The Process of Irony Comprehension in Young Children: Evidence from Monolingual and Bilingual Preschoolers

Authors: Natalia Banasik

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Comprehension of verbal irony is an example of pragmatic competence in understanding figurative language. The knowledge of how it develops may shed new light on the understanding of social and communicative competence that is crucial for one's effective functioning in the society. Researchers agree it is a competence that develops late in a child’s development. One of the abilities that seems crucial for irony comprehension is theory of mind (ToM), that is the ability to understand that others may have beliefs, desires and intentions different from one’s own. Although both theory of mind and irony comprehension require the ability to understand the figurative use of the false description of the reality, the exact relationship between them is still unknown. Also, even though irony comprehension in children has been studied for over thirty years, the results of the studies are inconsistent as to the age when this competence are acquired. The presented study aimed to answer questions about the developmental trajectories of irony comprehension and ascribing function to ironic utterances by preschool children. Specifically, we were interested in how it is related to the development of ToM and how comprehension of the function of irony changes with age. Data was collected from over 150 monolingual, Polish-speaking children and (so far) thirty bilingual children speaking Polish and English who live in the US. Four-, five- and six-year-olds were presented with a story comprehension task in the form of audio and visual stimuli programmed in the E-prime software (pre-recorded narrated stories, some of which included ironic utterances, and pictures accompanying the stories displayed on a touch screen). Following the presentation, the children were then asked to answer a series of questions. The questions checked the children’s understanding of the intended utterance meaning, evaluation of the degree to which it was funny and evaluation of how nice the speaker was. The children responded by touching the screen, which made it possible to measure reaction times. Additionally, the children were asked to explain why the speaker had uttered the ironic statement. Both quantitive and qualitative analyses were applied. The results of our study indicate that for irony recognition there is a significant difference among the three age groups, but what is new is that children as young as four do understand the real meaning behind the ironic statement as long as the utterance is not grammtically or lexically complex also, there is a clear correlation of ToM and irony comprehension. Although four-year olds and six-year olds understand the real meaning of the ironic utterance, it is not earlier than at the age of six when children start to explain the reason of using this marked form of expression. They talk about the speaker's intention to tell a joke, be funny, or to protect the listener's emotions. There are also some metalinguistic references, such as "mommy sometimes says things that don't make sense and this is called a metaphor".

Keywords: child's pragmatics, figurative speech, irony comprehension in children, theory of mind and irony

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250 Evolution of Antimicrobial Resistance in Shigella since the Turn of 21st Century, India

Authors: Neelam Taneja, Abhishek Mewara, Ajay Kumar

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Multidrug resistant shigellae have emerged as a therapeutic challenge in India. At our 2000 bed tertiary care referral centre in Chandigarh, North India, which caters to a large population of 7 neighboring states, antibiotic resistance in Shigella is being constantly monitored. Shigellae are isolated from 3 to 5% of all stool samples. In 1990 nalidixic acid was the drug of choice as 82%, and 63% of shigellae were resistant to ampicillin and cotrimoxazole respectively. Nalidixic acid resistance emerged in 1992 and rapidly increased from 6% during 1994-98 to 86% by the turn of 21st century. In the 1990s, the WHO recommended ciprofloxacin as the drug of choice for empiric treatment of shigellosis in view of the existing high level resistance to agents like chloramphenicol, ampicillin, cotrimoxazole and nalidixic acid. First resistance to ciprofloxacin in S. flexneri at our centre appeared in 2000 and rapidly rose to 46% in 2007 (MIC>4mg/L). In between we had an outbreak of ciprofloxacin resistant S.dysenteriae serotype 1 in 2003. Therapeutic failures with ciprofloxacin occurred with both ciprofloxacin-resistant S. dysenteriae and ciprofloxacin-resistant S. flexneri. The severity of illness was more with ciprofloxacin-resistant strains. Till 2000, elsewhere in the world ciprofloxacin resistance in S. flexneri was sporadic and uncommon, though resistance to co-trimoxazole and ampicillin was common and in some areas resistance to nalidixic acid had also emerged. Fluoroquinolones due to extensive use and misuse for many other illnesses in our region are thus no longer the preferred group of drugs for managing shigellosis in India. WHO presently recommends ceftriaxone and azithromycin as alternative drugs to fluoroquinolone-resistant shigellae, however, overreliance on this group of drugs also seems to soon become questionable considering the emerging cephalosporin-resistant shigellae. We found 15.1% of S. flexneri isolates collected over a period of 9 years (2000-2009) resistant to at least one of the third-generation cephalosporins (ceftriaxone/cefotaxime). The first isolate showing ceftriaxone resistance was obtained in 2001, and we have observed an increase in number of isolates resistant to third generation cephalosporins in S. flexneri 2005 onwards. This situation has now become a therapeutic challenge in our region. The MIC values for Shigella isolates revealed a worrisome rise for ceftriaxone (MIC90:12 mg/L) and cefepime (MIC90:8 mg/L). MIC values for S. dysenteriae remained below 1 mg/L for ceftriaxone, however for cefepime, the MIC90 has raised to 4 mg/L. These infections caused by ceftriaxone-resistant S. flexneri isolates were successfully treated by azithromycin at our center. Most worrisome development in the present has been the emergence of DSA(Decreased susceptibility to azithromycin) which surfaced in 2001 and has increased from 4.3% till 2011 to 34% thereafter. We suspect plasmid-mediated resistance as we detected qnrS1-positive Shigella for the first time from the Indian subcontinent in 2 strains from 2010, indicating a relatively new appearance of this PMQR determinant among Shigella in India. This calls for a continuous and strong surveillance of antibiotic resistance across the country. The prevention of shigellosis by developing cost-effective vaccines is desirable as it will substantially reduce the morbidity associated with diarrhoea in the country

Keywords: Shigella, antimicrobial, resistance, India

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249 Reconceptualising the Voice of Children in Child Protection

Authors: Sharon Jackson, Lynn Kelly

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This paper proposes a conceptual review of the interdisciplinary literature which has theorised the concept of ‘children’s voices’. The primary aim is to identify and consider the theoretical relevance of conceptual thought on ‘children’s voices’ for research and practice in child protection contexts. Attending to the ‘voice of the child’ has become a core principle of social work practice in contemporary child protection contexts. Discourses of voice permeate the legislative, policy and practice frameworks of child protection practices within the UK and internationally. Voice is positioned within a ‘child-centred’ moral imperative to ‘hear the voices’ of children and take their preferences and perspectives into account. This practice is now considered to be central to working in a child-centered way. The genesis of this call to voice is revealed through sociological analysis of twentieth-century child welfare reform as rooted inter alia in intersecting political, social and cultural discourses which have situated children and childhood as cites of state intervention as enshrined in the 1989 United Nations Convention on the Rights of the Child ratified by the UK government in 1991 and more specifically Article 12 of the convention. From a policy and practice perspective, the professional ‘capturing’ of children’s voices has come to saturate child protection practice. This has incited a stream of directives, resources, advisory publications and ‘how-to’ guides which attempt to articulate practice methods to ‘listen’, ‘hear’ and above all – ‘capture’ the ‘voice of the child’. The idiom ‘capturing the voice of the child’ is frequently invoked within the literature to express the requirements of the child-centered practice task to be accomplished. Despite the centrality of voice, and an obsession with ‘capturing’ voices, evidence from research, inspection processes, serious case reviews, child abuse and death inquires has consistently highlighted professional neglect of ‘the voice of the child’. Notable research studies have highlighted the relative absence of the child’s voice in social work assessment practices, a troubling lack of meaningful engagement with children and the need to more thoroughly examine communicative practices in child protection contexts. As a consequence, the project of capturing ‘the voice of the child’ has intensified, and there has been an increasing focus on developing methods and professional skills to attend to voice. This has been guided by a recognition that professionals often lack the skills and training to engage with children in age-appropriate ways. We argue however that the problem with ‘capturing’ and [re]representing ‘voice’ in child protection contexts is, more fundamentally, a failure to adequately theorise the concept of ‘voice’ in the ‘voice of the child’. For the most part, ‘The voice of the child’ incorporates psychological conceptions of child development. While these concepts are useful in the context of direct work with children, they fail to consider other strands of sociological thought, which position ‘the voice of the child’ within an agentic paradigm to emphasise the active agency of the child.

Keywords: child-centered, child protection, views of the child, voice of the child

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248 In-situ Acoustic Emission Analysis of a Polymer Electrolyte Membrane Water Electrolyser

Authors: M. Maier, I. Dedigama, J. Majasan, Y. Wu, Q. Meyer, L. Castanheira, G. Hinds, P. R. Shearing, D. J. L. Brett

Abstract:

Increasing the efficiency of electrolyser technology is commonly seen as one of the main challenges on the way to the Hydrogen Economy. There is a significant lack of understanding of the different states of operation of polymer electrolyte membrane water electrolysers (PEMWE) and how these influence the overall efficiency. This in particular means the two-phase flow through the membrane, gas diffusion layers (GDL) and flow channels. In order to increase the efficiency of PEMWE and facilitate their spread as commercial hydrogen production technology, new analytic approaches have to be found. Acoustic emission (AE) offers the possibility to analyse the processes within a PEMWE in a non-destructive, fast and cheap in-situ way. This work describes the generation and analysis of AE data coming from a PEM water electrolyser, for, to the best of our knowledge, the first time in literature. Different experiments are carried out. Each experiment is designed so that only specific physical processes occur and AE solely related to one process can be measured. Therefore, a range of experimental conditions is used to induce different flow regimes within flow channels and GDL. The resulting AE data is first separated into different events, which are defined by exceeding the noise threshold. Each acoustic event consists of a number of consequent peaks and ends when the wave diminishes under the noise threshold. For all these acoustic events the following key attributes are extracted: maximum peak amplitude, duration, number of peaks, peaks before the maximum, average intensity of a peak and time till the maximum is reached. Each event is then expressed as a vector containing the normalized values for all criteria. Principal Component Analysis is performed on the resulting data, which orders the criteria by the eigenvalues of their covariance matrix. This can be used as an easy way of determining which criteria convey the most information on the acoustic data. In the following, the data is ordered in the two- or three-dimensional space formed by the most relevant criteria axes. By finding spaces in the two- or three-dimensional space only occupied by acoustic events originating from one of the three experiments it is possible to relate physical processes to certain acoustic patterns. Due to the complex nature of the AE data modern machine learning techniques are needed to recognize these patterns in-situ. Using the AE data produced before allows to train a self-learning algorithm and develop an analytical tool to diagnose different operational states in a PEMWE. Combining this technique with the measurement of polarization curves and electrochemical impedance spectroscopy allows for in-situ optimization and recognition of suboptimal states of operation.

Keywords: acoustic emission, gas diffusion layers, in-situ diagnosis, PEM water electrolyser

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247 Carbon Nanotubes (CNTs) as Multiplex Surface Enhanced Raman Scattering Sensing Platforms

Authors: Pola Goldberg Oppenheimer, Stephan Hofmann, Sumeet Mahajan

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Owing to its fingerprint molecular specificity and high sensitivity, surface-enhanced Raman scattering (SERS) is an established analytical tool for chemical and biological sensing capable of single-molecule detection. A strong Raman signal can be generated from SERS-active platforms given the analyte is within the enhanced plasmon field generated near a noble-metal nanostructured substrate. The key requirement for generating strong plasmon resonances to provide this electromagnetic enhancement is an appropriate metal surface roughness. Controlling nanoscale features for generating these regions of high electromagnetic enhancement, the so-called SERS ‘hot-spots’, is still a challenge. Significant advances have been made in SERS research, with wide-ranging techniques to generate substrates with tunable size and shape of the nanoscale roughness features. Nevertheless, the development and application of SERS has been inhibited by the irreproducibility and complexity of fabrication routes. The ability to generate straightforward, cost-effective, multiplex-able and addressable SERS substrates with high enhancements is of profound interest for miniaturised sensing devices. Carbon nanotubes (CNTs) have been concurrently, a topic of extensive research however, their applications for plasmonics has been only recently beginning to gain interest. CNTs can provide low-cost, large-active-area patternable substrates which, coupled with appropriate functionalization capable to provide advanced SERS-platforms. Herein, advanced methods to generate CNT-based SERS active detection platforms will be discussed. First, a novel electrohydrodynamic (EHD) lithographic technique will be introduced for patterning CNT-polymer composites, providing a straightforward, single-step approach for generating high-fidelity sub-micron-sized nanocomposite structures within which anisotropic CNTs are vertically aligned. The created structures are readily fine-tuned, which is an important requirement for optimizing SERS to obtain the highest enhancements with each of the EHD-CNTs individual structural units functioning as an isolated sensor. Further, gold-functionalized VACNTFs are fabricated as SERS micro-platforms. The dependence on the VACNTs’ diameters and density play an important role in the Raman signal strength, thus highlighting the importance of structural parameters, previously overlooked in designing and fabricating optimized CNTs-based SERS nanoprobes. VACNTs forests patterned into predesigned pillar structures are further utilized for multiplex detection of bio-analytes. Since CNTs exhibit electrical conductivity and unique adsorption properties, these are further harnessed in the development of novel chemical and bio-sensing platforms.

Keywords: carbon nanotubes (CNTs), EHD patterning, SERS, vertically aligned carbon nanotube forests (VACNTF)

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246 A Comparative Semantic Network Study between Chinese and Western Festivals

Authors: Jianwei Qian, Rob Law

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With the expansion of globalization and the increment of market competition, the festival, especially the traditional one, has demonstrated its vitality under the new context. As a new tourist attraction, festivals play a critically important role in promoting the tourism economy, because the organization of a festival can engage more tourists, generate more revenues and win a wider media concern. However, in the current stage of China, traditional festivals as a way to disseminate national culture are undergoing the challenge of foreign festivals and the related culture. Different from those special events created solely for developing economy, traditional festivals have their own culture and connotation. Therefore, it is necessary to conduct a study on not only protecting the tradition, but promoting its development as well. This study conducts a comparative study of the development of China’s Valentine’s Day and Western Valentine’s Day under the Chinese context and centers on newspaper reports in China from 2000 to 2016. Based on the literature, two main research focuses can be established: one is concerned about the festival’s impact and the other is about tourists’ motivation to engage in a festival. Newspaper reports serve as the research discourse and can help cover the two focal points. With the assistance of content mining techniques, semantic networks for both Days are constructed separately to help depict the status quo of these two festivals in China. Based on the networks, two models are established to show the key component system of traditional festivals in the hope of perfecting the positive role festival tourism plays in the promotion of economy and culture. According to the semantic networks, newspaper reports on both festivals have similarities and differences. The difference is mainly reflected in its cultural connotation, because westerners and Chinese may show their love in different ways. Nevertheless, they share more common points in terms of economy, tourism, and society. They also have a similar living environment and stakeholders. Thus, they can be promoted together to revitalize some traditions in China. Three strategies are proposed to realize the aforementioned aim. Firstly, localize international festivals to suit the Chinese context to make it function better. Secondly, facilitate the internationalization process of traditional Chinese festivals to receive more recognition worldwide. Finally, allow traditional festivals to compete with foreign ones to help them learn from each other and elucidate the development of other festivals. It is believed that if all these can be realized, not only the traditional Chinese festivals can obtain a more promising future, but foreign ones are the same as well. Accordingly, the paper can contribute to the theoretical construction of festival images by the presentation of the semantic network. Meanwhile, the identified features and issues of festivals from two different cultures can enlighten the organization and marketing of festivals as a vital tourism activity. In the long run, the study can enhance the festival as a key attraction to keep the sustainable development of both the economy and the society.

Keywords: Chinese context, comparative study, festival tourism, semantic network analysis, valentine’s day

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245 Design and Application of a Model Eliciting Activity with Civil Engineering Students on Binomial Distribution to Solve a Decision Problem Based on Samples Data Involving Aspects of Randomness and Proportionality

Authors: Martha E. Aguiar-Barrera, Humberto Gutierrez-Pulido, Veronica Vargas-Alejo

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Identifying and modeling random phenomena is a fundamental cognitive process to understand and transform reality. Recognizing situations governed by chance and giving them a scientific interpretation, without being carried away by beliefs or intuitions, is a basic training for citizens. Hence the importance of generating teaching-learning processes, supported using technology, paying attention to model creation rather than only executing mathematical calculations. In order to develop the student's knowledge about basic probability distributions and decision making; in this work a model eliciting activity (MEA) is reported. The intention was applying the Model and Modeling Perspective to design an activity related to civil engineering that would be understandable for students, while involving them in its solution. Furthermore, the activity should imply a decision-making challenge based on sample data, and the use of the computer should be considered. The activity was designed considering the six design principles for MEA proposed by Lesh and collaborators. These are model construction, reality, self-evaluation, model documentation, shareable and reusable, and prototype. The application and refinement of the activity was carried out during three school cycles in the Probability and Statistics class for Civil Engineering students at the University of Guadalajara. The analysis of the way in which the students sought to solve the activity was made using audio and video recordings, as well as with the individual and team reports of the students. The information obtained was categorized according to the activity phase (individual or team) and the category of analysis (sample, linearity, probability, distributions, mechanization, and decision-making). With the results obtained through the MEA, four obstacles have been identified to understand and apply the binomial distribution: the first one was the resistance of the student to move from the linear to the probabilistic model; the second one, the difficulty of visualizing (infering) the behavior of the population through the sample data; the third one, viewing the sample as an isolated event and not as part of a random process that must be viewed in the context of a probability distribution; and the fourth one, the difficulty of decision-making with the support of probabilistic calculations. These obstacles have also been identified in literature on the teaching of probability and statistics. Recognizing these concepts as obstacles to understanding probability distributions, and that these do not change after an intervention, allows for the modification of these interventions and the MEA. In such a way, the students may identify themselves the erroneous solutions when they carrying out the MEA. The MEA also showed to be democratic since several students who had little participation and low grades in the first units, improved their participation. Regarding the use of the computer, the RStudio software was useful in several tasks, for example in such as plotting the probability distributions and to exploring different sample sizes. In conclusion, with the models created to solve the MEA, the Civil Engineering students improved their probabilistic knowledge and understanding of fundamental concepts such as sample, population, and probability distribution.

Keywords: linear model, models and modeling, probability, randomness, sample

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244 Antimicrobial Activities of Lactic Acid Bacteria from Fermented Foods and Probiotic Products

Authors: Alec Chabwinja, Cannan Tawonezvi, Jerneja Vidmar, Constance Chingwaru, Walter Chingwaru

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Objective: To evaluate the potential of commercial fermented / probiotic products available in Zimbabwe or internationally, and strains of Lactobacillus plantarum (L. plantarum) as prophylaxis and therapy against diarrhoeal and sexually transmitted infections. Methods: The antimicrobial potential of cultures of lactobacilli enriched from 4 Zimbabwean commercial food/beverage products, namely Dairibord Lacto sour milk (DLSM), Probrand sour milk (PSM), Kefalos Vuka cheese (KVC) and Chibuku opaque beer (COB); three probiotic products obtainable in Europe and internationally; and four strains of L. plantarum obtained from Balkan traditional cheeses and Zimbabwean foods against clinical strains of Escherichia coli (E. coli) and non-clinical strains of Candida albicans and Rhodotorula spp. was assayed using the well diffusion method. Three commercial Agar diffusion assay and a competitive exclusion assay were carried out on Mueller-Hinton agar. Results: Crude cultures of putative lactobacillus strains obtained from Zimbabwean dairy products (Probrand sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer) exhibited significantly greater antimicrobial activities against clinical strains of E. coli than strains of L. plantarum isolated from Balkan cheeses (CLP1, CLP2 or CLP3) or crude microbial cultures from commercial paediatric probiotic products (BG, PJ and PL) of a culture of Lactobacillus rhamnosus LGG (p < 0.05). Furthermore, the following has high antifungal activities against the two yeasts: supernatant-free microbial pellet (SFMP) from an extract of M. azedarach leaves (27mm ± 2.5) > cell-free culture supernatants (CFCS) from Maaz Dairy sour milk and Mnandi sour milk (approximately 26mm ± 1.8) > CFCS and SFMP from Amansi hodzeko (25mm ± 1.5) > CFCS from Parinari curatellifolia fruit (24mm ± 1.5), SFMP from P. curatellifolia fruit (24mm ± 1.4) and SFMP from mahewu (20mm ± 1.5). These cultures also showed high tolerance to acidic conditions (~pH4). Conclusions: The putative lactobacilli from four commercial Zimbabwean dairy products (Probrand sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer), and three strains of L. plantarum from Balkan cheeses (CLP1, CLP2 or CLP3) exhibited high antibacterial activities, while Maaz Dairy sour-, Mnandi sour- and Amansi hodzeko milk products had high antifungal activities. Our selection of Zimbabwean probiotic products has potential for further development into probiotic products for use in the control diarrhea caused by pathogenic strains of E. coli or yeast infections. Studies to characterise the probiotic potential of the live cultures in the products are underway.

Keywords: lactic acid bacteria, Staphylococcus aureus, Streptococcus spp, yeast, inhibition, acid tolerance

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243 Understanding and Measuring Stigma, Barriers and Attitudes Associated with Seeking Psychological Help Among Young Adults in Czech Republic

Authors: Tereza Hruskova

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200 million people globally experience serious mental health problems, and only one third seek professional help, and help-seeking is described as a last resort. Adolescents and young adults have a high prevalence of mental illness. Mental stigma is a key element in the decision to seek help and is divided into (i) self-stigma (self-stigmatization), including internal beliefs, low self-esteem, and lower quality of life, and (ii) public stigma (social stigma) containing stereotypes, beliefs and society's disapproval of help-seeking having a negative effect on help-seeking and our attitudes. Previous research has mainly focused on examining the construct of help seeking, avoidance, and delaying separately and trying to find out why people do not seek help in time and what obstacles stand in the way. Barriers are not static and may change over time and the stage of help-seeking. Attitudes are closely related to self-stigma and social stigma and predict whether a person will seek help. Barriers (stigmatization, a sense of humiliation, insufficient recognition of the problem, preferences, solving it alone, and distrust of a professional) and facilitators (previous experience with mental problems, social support, and help from others) are factors influencing help-seeking. The current research on the Czech population of young adults responds to the gap between a person with mental health problems and actually seeking professional help. The aim of the study is to describe in detail the individual constructs and factors, to understand the person seeking help, and to define possible obstacles on this path of seeking help. A sample of approximately 250 participants (age 18-35) would take part in the online questionnaire, conducted in May-June 2023, and would be administered a demographic questionnaire and four scales measuring attitudes (Attitudes Toward Seeking Professional Psychological Help – Short form), barriers (Barrier to Help Seeking Scale), self-stigma (Self Stigma of Seeking Help) and stigmatization (Perceptions of Stigmatization by Others for seeking help). Firstly, all four scales would be translated into the Czech language. The aim is (I) to determine the validity and reliability of the Czech translation of the scales, (II) to examine the factors of the scales on the Czech population and compare them retrospectively with the results of reliability and validity from the original language of the scales and (III) to examine the connections between attitudes towards seeking, avoidance or delaying the search for professional psychological help due to the demographic and individual differences of the participants, barriers, self-stigmatization and social stigmatization. We expect to carry out the first study on the given topic in the Czech Republic, to identify and better understand the factors leading to the avoidance of seeking professional help and to reveal the relationships between stigmatization, attitudes and barriers leading to the avoidance or postponement of seeking professional help. The belief is to find out whether the Czech population of young adults differs from the data found on the foreign population in individual constructs, as cultural differences in individual countries were found.

Keywords: mental health, stigma, problems, seeking psychological help

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242 The Production of Biofertilizer from Naturally Occurring Microorganisms by Using Nuclear Technologies

Authors: K. S. Al-Mugren, A. Yahya, S. Alodah, R. Alharbi, S. H. Almsaid , A. Alqahtani, H. Jaber, A. Basaqer, N. Alajra, N. Almoghati, A. Alsalman, Khalid Alharbi

Abstract:

Context: The production of biofertilizers from naturally occurring microorganisms is an area of research that aims to enhance agricultural practices by utilizing local resources. This research project focuses on isolating and screening indigenous microorganisms with PK-fixing and phosphate solubilizing characteristics from local sources. Research Aim: The aim of this project is to develop a biofertilizer product using indigenous microorganisms and composted agro waste as a carrier. The objective is to enhance crop productivity and soil fertility through the application of biofertilizers. Methodology: The research methodology includes several key steps. Firstly, indigenous microorganisms will be isolated from local resources using the ten-fold serial dilutions technique. Screening assays will be conducted to identify microorganisms with phosphate solubilizing and PK-fixing activities. Agro-waste materials will be collected from local agricultural sources, and composting experiments will be conducted to convert them into organic matter-rich compost. Physicochemical analysis will be performed to assess the composition of the composted agro-waste. Gamma and X-ray irradiation will be used to sterilize the carrier material. The sterilized carrier will be tested for sterility using the ten-fold serial dilutions technique. Finally, selected indigenous microorganisms will be developed into biofertilizer products. Findings: The research aims to find suitable indigenous microorganisms with phosphate solubilizing and PK-fixing characteristics for biofertilizer production. Additionally, the research aims to assess the suitability of composted agro waste as a carrier for biofertilizers. The impact of gamma irradiation sterilization on pathogen elimination will also be investigated. Theoretical Importance: This research contributes to the understanding of utilizing indigenous microorganisms and composted agro waste for biofertilizer production. It expands knowledge on the potential benefits of biofertilizers in enhancing crop productivity and soil fertility. Data Collection and Analysis Procedures: The data collection process involves isolating indigenous microorganisms, conducting screening assays, collecting and composting agro waste, analyzing the physicochemical composition of composted agro waste, and testing carrier sterilization. The analysis procedures include assessing the abilities of indigenous microorganisms, evaluating the composition of composted agro waste, and determining the sterility of the carrier material. Conclusion: The research project aims to develop biofertilizer products using indigenous microorganisms and composted agro waste as a carrier. Through the isolation and screening of indigenous microorganisms, the project aims to enhance crop productivity and soil fertility by utilizing local resources. The research findings will contribute to the understanding of the suitability of composted agro waste as a carrier and the efficacy of gamma irradiation sterilization. The research outcomes will have theoretical importance in the field of biofertilizer production and agricultural practices.

Keywords: biofertilizer, microorganisms, agro waste, nuclear technologies

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241 Self-Education, Recognition and Well-Being Insights into Qualitative-Reconstructive Educational Research on the Value of Non-formal Education in the Adolescence

Authors: Sandra Biewers Grimm

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International studies such as Pisa have shown an increasing social inequality in the education system, which is determined in particular by social origin and migration status. This is especially the case in the Luxembourg school system, which creates challenges for many young people due to the multilingualism in the country. While the international and also the national debate on education in the immediate aftermath of the publications of the Pisa results mainly focused on the further development of school-based learning venues and formal educational processes, it initially remained largely unclear what role exactly out-of-school learning venues and non-formal and informal learning processes could play in this further development. This has changed in the meantime. Both in the political discourses and in the scientific disciplines, those voices have become louder that draw attention to the important educational function and the enormous educational potential of out-of-school learning places as a response to the crisis of the formal education system and more than this. Youth work as an actor and approach of non-formal education is particularly in demand here. Due to its principles of self-education, participation and openness, it is considered to have a special potential in supporting the acquisition of important key competencies. In this context, the study "Educational experiences in non-formal settings" at CCY takes a differentiated look behind the scenes of education-oriented youth work and describes on the basis of empirical data what and how young people learn in youth centers and which significance they attach to these educational experiences for their subjective life situation. In this sense, the aim of the study is to reconstruct the subjective educational experiences of young people in Open Youth Work as well as to explore the value that these experiences have for young people. In doing so, it enables scientifically founded conclusions about the educational potential of youth work from the user's perspective. Initially, the study focuses on defining the concept of education in the context of non-formal education and thus sets a theoretical framework for the empirical analysis. This socio-educational term of education differs from the relevant conception of education in curricular, formal education as the acquisition of knowledge. It also differs from the operationalization of education as competence, or the differentiation into cultural, social and personal or into factual, social or methodological competence, which is often used in the European context and which has long been interpreted as a "social science reading of the question of education" (XX). Now the aim is to define a "broader" concept of education that goes beyond the normative and educational policy dimensions of a "non-formal education" and includes the classical socio-educational dimensions. Furthermore, the study works with different methods of empirical social research: In addition to ethnographic observation and an online survey, group discussions were conducted with the young people. The presentation gives an insight into the context, the methodology and the results of this study.

Keywords: non-formal education, youth research, qualitative research, educational theory

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240 Phylogenetic Inferences based on Morphoanatomical Characters in Plectranthus esculentus N. E. Br. (Lamiaceae) from Nigeria

Authors: Otuwose E. Agyeno, Adeniyi A. Jayeola, Bashir A. Ajala

Abstract:

P. esculentus is indigenous to Nigeria yet no wild relation has been encountered or reported. This has made it difficult to establish proper lineages between the varieties and landraces under cultivation. The present work is the first to determine the apormophy of 135 morphoanatomical characters in organs of 46 accessions drawn from 23 populations of this species based on dicta. The character states were coded in accession x character-state matrices and only 83 were informative and utilised for neighbour joining clustering based on euclidean values, and heuristic search in parsimony analysis using PAST ver. 3.15 software. Compatibility and evolutionary trends between accessions were then explored from values and diagrams produced. The low consistency indices (CI) recorded support monophyly and low homoplasy in this taxon. Agglomerative schedules based on character type and source data sets divided the accessions into mainly 3 clades, each of complexes of accessions. Solenostemon rotundifolius (Poir) J.K Morton was the outgroup (OG) used, and it occurred within the largest clades except when the characters were combined in a data set. The OG showed better compatibility with accessions of populations of landrace Isci, and varieties Riyum and Long’at. Otherwise, its aerial parts are more consistent with those of accessions of variety Bebot. The highly polytomous clades produced due to anatomical data set may be an indication of how stable such characters are in this species. Strict consensus trees with more than 60 nodes outputted showed that the basal nodes were strongly supported by 3 to 17 characters across the data sets, suggesting that populations of this species are more alike. The OG was clearly the first diverging lineage and closely related to accessions of landrace Gwe and variety Bebot morphologically, but different from them anatomically. It was also distantly related to landrace Fina and variety Long’at in terms of root, stem and leaf structural attributes. There were at least 5 other clades with each comprising of complexes of accessions from different localities and terrains within the study area. Spherical stem in cross section, size of vascular bundles at the stem corners as well as the alternate and whorl phyllotaxy are attributes which may have facilitated each other’s evolution in all accessions of the landrace Gwe, and they may be innovative since such states are not characteristic of the larger Lamiaceae, and Plectranthus L’Her in particular. In conclusion, this study has provided valuable information about infraspecific diversity in this taxon. It supports recognition of the varietal statuses accorded to populations of P. esculentus, as well as the hypothesis that the wild gene might have been distributed on the Jos Plateau. However, molecular characterisation of accessions of populations of this species would resolve this problem better.

Keywords: clustering, lineage, morphoanatomical characters, Nigeria, phylogenetics, Plectranthus esculentus, population

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239 Generating a Multiplex Sensing Platform for the Accurate Diagnosis of Sepsis

Authors: N. Demertzis, J. L. Bowen

Abstract:

Sepsis is a complex and rapidly evolving condition, resulting from uncontrolled prolonged activation of host immune system due to pathogenic insult. The aim of this study is the development of a multiplex electrochemical sensing platform, capable of detecting both pathogen associated and host immune markers to enable the rapid and definitive diagnosis of sepsis. A combination of aptamers and molecular imprinting approaches have been employed to generate sensing systems for lipopolysaccharide (LPS), c-reactive protein (CRP) and procalcitonin (PCT). Gold working electrodes were mechanically polished and electrochemically cleaned with 0.1 M sulphuric acid using cyclic voltammetry (CV). Following activation, a self-assembled monolayer (SAM) was generated, by incubating the electrodes with a thiolated anti-LPS aptamer / dithiodibutiric acid (DTBA) mixture (1:20). 3-aminophenylboronic acid (3-APBA) in combination with the anti-LPS aptamer was used for the development of the hybrid molecularly imprinted sensor (apta-MIP). Aptasensors, targeting PCT and CRP were also fabricated, following the same approach as in the case of LPS, with mercaptohexanol (MCH) replacing DTBA. In the case of the CRP aptasensor, the SAM was formed following incubation of a 1:1 aptamer: MCH mixture. However, in the case of PCT, the SAM was formed with the aptamer itself, with subsequent backfilling with 1 μM MCH. The binding performance of all systems has been evaluated using electrochemical impedance spectroscopy. The apta-MIP’s polymer thickness is controlled by varying the number of electropolymerisation cycles. In the ideal number of polymerisation cycles, the polymer must cover the electrode surface and create a binding pocket around LPS and its aptamer binding site. Less polymerisation cycles will create a hybrid system which resembles an aptasensor, while more cycles will be able to cover the complex and demonstrate a bulk polymer-like behaviour. Both aptasensor and apta-MIP were challenged with LPS and compared to conventional imprinted (absence of aptamer from the binding site, polymer formed in presence of LPS) and non-imprinted polymers (NIPS, absence of LPS whilst hybrid polymer is formed). A stable LPS aptasensor, capable of detecting down to 5 pg/ml of LPS was generated. The apparent Kd of the system was estimated at 17 pM, with a Bmax of approximately 50 pM. The aptasensor demonstrated high specificity to LPS. The apta-MIP demonstrated superior recognition properties with a limit of detection of 1 fg/ml and a Bmax of 100 pg/ml. The CRP and PCT aptasensors were both able to detect down to 5 pg/ml. Whilst full binding performance is currently being evaluated, there is none of the sensors demonstrate cross-reactivity towards LPS, CRP or PCT. In conclusion, stable aptasensors capable of detecting LPS, PCT and CRP at low concentrations have been generated. The realisation of a multiplex panel such as described herein, will effectively contribute to the rapid, personalised diagnosis of sepsis.

Keywords: aptamer, electrochemical impedance spectroscopy, molecularly imprinted polymers, sepsis

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238 Teaching Young Children Social and Emotional Learning through Shared Book Reading: Project GROW

Authors: Stephanie Al Otaiba, Kyle Roberts

Abstract:

Background and Significance Globally far too many students read below grade level; thus improving literacy outcomes is vital. Research suggests that non-cognitive factors, including Social and Emotional Learning (SEL) are linked to success in literacy outcomes. Converging evidence exists that early interventions are more effective than later remediation; therefore teachers need strategies to support early literacy while developing students’ SEL and their vocabulary, or language, for learning. This presentation describe findings from a US federally-funded project that trained teachers to provide an evidence-based read-aloud program for young children, using commercially available books with multicultural characters and themes to help their students “GROW”. The five GROW SEL themes include: “I can name my feelings”, “I can learn from my mistakes”, “I can persist”, “I can be kind to myself and others”, and “I can work toward and achieve goals”. Examples of GROW vocabulary (from over 100 words taught across the 5 units) include: emotions, improve, resilient, cooperate, accomplish, responsible, compassion, adapt, achieve, analyze. Methodology This study used a mixed methods research design, with qualitative methods to describe data from teacher feedback surveys (regarding satisfaction, feasibility), observations of fidelity of implementation, and with quantitative methods to assess the effect sizes for student vocabulary growth. GROW Intervention and Teacher Training Procedures Researchers trained classroom teachers to implement GROW. Each thematic unit included four books, vocabulary cards with images of the vocabulary words, and scripted lessons. Teacher training included online and in-person training; researchers incorporated virtual reality videos of instructors with child avatars to model lessons. Classroom teachers provided 2-3 20 min lessons per week ranging from short-term (8 weeks) to longer-term trials for up to 16 weeks. Setting and Participants The setting for the study included two large urban charter schools in the South. Data was collected across two years; during the first year, participants included 7 kindergarten teachers and 108 and the second year involved an additional set of 5 kindergarten and first grade teachers and 65 students. Initial Findings The initial qualitative findings indicate teachers reported the lessons to be feasible to implement and they reported that students enjoyed the books. Teachers found the vocabulary words to be challenging and important. They were able to implement lessons with fidelity. Quantitative analyses of growth for each taught word suggest that students’ growth on taught words ranged from large (ES = .75) to small (<.20). Researchers will contrast the effects for more and less successful books within the GROW units. Discussion and Conclusion It is feasible for teachers of young students to effectively teach SEL vocabulary and themes during shared book reading. Teachers and students enjoyed the books and students demonstrated growth on taught vocabulary. Researchers will discuss implications of the study and about the GROW program for researchers in learning sciences, will describe some limitations about research designs that are inherent in school-based research partnerships, and will provide some suggested directions for future research and practice.

Keywords: early literacy, learning science, language and vocabulary, social and emotional learning, multi-cultural

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237 Antifungal Activity of Processed Sulfur Solution as Potential Eco-Friendly Disinfectant against Saprolegnia parasitica and Its Safety in Freshwater-Farmed Fish

Authors: Hye-Hyun Lee, Hyo-Kon Chun, Kyung-Hee Kim Kim, Mi-Hee Kim, Saet-Byul Chu, Sang-Jong Lee, Seung-Hyeop Lee, Seung-Won Yi

Abstract:

Some chemicals such as malachite green, methylene blue, and copper sulfate had been used frequently as disinfectants controlling fungal infection in aquaculture. However, their carcinogenicity, mutagenicity and teratogenicity were reported in mammals. After their accumulation in food fish and its consumers was confirmed, concerns about public health has resulted in enhanced monitoring and increased demand for eco-friendly treatments. Therefore, this study aimed to evaluate safety to fish and efficacy of sulfur solution processed by effective microorganisms (EM-PSS) against Saprolegnia parasitica, for use of a potential aquatic fungicidal disinfectant. The natural sulfur purchased from Kawah Ijen volcano, East Java, Indonesia was processed by the liquid mixture consisting of following twelve effective microorganisms (Rapha-el®; Lbiotech, Jeonnam, Korea), Lactobacillus parafarraginis, L. paracasei, L. harbinensis, L. buchneri, L. perolens, L. rhamnosus, L. vaccinostercus, Acetobacter lovaniensis, A. peroxydans, Pichia fermentans, Candida ethanolica, Saccharomycopsis schoenii isolated from fermentation process of oriental medicinal herbs including green tea, privet, and puer tea. The material was applied to in vitro antifungal activity test for Saprolegnia parasitica using agar dilution method. In addition, an acute toxicity test was performed on carp (Cyprinus carpio), eel (Anguilla japonica), and mud fish (Misgurnus mizolepis) for 96 hours. After three species of fish (n=15) were accustomed to experimental water environment for three days, the EM-PSS was added to each tank as final concentrations to be 0 to 500 ppm. The fish were taken into necropsy, and the histological sections of the gill, liver, and spleen were counter-stained with hematoxylin and eosin (H-E). And hence, no observed effect concentration (NOEC) of the solution was used for taking a medicinal bath for mudfish infected by Saprolegnia parasitica in practice. The result of in vitro antifungal activity test showed the growth inhibition of the fungus at 100 ppm, which and the lower concentrations occurred no fatal case in any fish species tested until the end of the examination. The 125 ppm of the solution, however, resulted in 13.3 %, 13.3 %, and 6.3 % of mortality in carp, eel, and mudfish, respectively. But both 250 and 500 ppm of the solution leaded lethality to all population of each fish species within 24 hours. Besides, H-E staining also showed no specific evidence for toxicity in fish at lesser than 100 ppm of EM-PSS. On the other hand, as a result of field application of the solution, no growth of fungal mycelium was found in fish bodies from gross observation 5 days post treatment. In conclusion, 100ppm of EM-PSS resulted in inhibition and treatment of Saprolegnia parasitica infection. In addition, the use of EM-PSS lower than 100 ppm is safe for fish. Therefore, EM-PSS could be used as aquatic fungicide, and also may be possible to be a potential eco-friendly disinfectant in aquaculture.

Keywords: antifungal activity, effective microorganism, toxicity, saprolegnia, processed sulfur solution

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236 Isolation and Characterization of a Narrow-Host Range Aeromonas hydrophila Lytic Bacteriophage

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

Abstract:

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

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

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