Search results for: dfi (dna fragmentation assay) (scd-sperm chromatin dispersion).art (artificial reproductive technology)

Authors: Jiraporn Weenuttranon

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The objectives of the consumer behavior of bakery products in Bangkok are to study consumer behavior of the bakery product, to study the essential factors that could possibly affect the consumer behavior and to study recommendations for the development of the bakery products. This research is a survey research. Populations are buyer’s bakery products in Bangkok. The probability sample size is 400. The research uses a questionnaire for self-learning by using information technology. The researcher created a reliability value at 0.71 levels of significance. The data analysis will be done by using the percentage, mean, and standard deviation and testing the hypotheses by using chi-square.

Keywords: consumer, behavior, bakery, standard deviation

Procedia PDF Downloads 458

Authors: Manahil Alfuraydan, Jodie Croxall, Lisa Hurt, Mike Kerr, Sinead Brophy

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Autism Spectrum Disorder (ASD) is a developmental condition characterised by impairment in terms of social communication, social interaction, and a repetitive or restricted pattern of interest, behaviour, and activity. There is a significant delay between seeking help and a confirmed diagnosis of ASD. This may result in delay in receiving early intervention services, which are critical for positive outcomes. The long wait times also cause stress for the individuals and their families. Telehealth potentially offers a way of improving the diagnostic pathway for ASD. This review of the literature aims to examine which telehealth approaches have been used in the diagnosis and assessment of autism in children and adults, whether they are feasible and acceptable, and how they compare with face-to-face diagnosis and assessment methods. A comprehensive search of following databases- MEDLINE, CINAHL Plus with Full text, Business Sources Complete, Web of Science, Scopus, PsycINFO and trail and systematic review databases including Cochrane Library, Health Technology Assessment, Database of Abstracts and Reviews of Effectiveness and NHS Economic Evaluation was conducted, combining the terms of autism and telehealth from 2000 to 2018. A total of 10 studies were identified for inclusion in the review. This review of the literature found there to be two methods of using telehealth: (a) video conferencing to enable teams in different areas to consult with the families and to assess the child/adult in real time and (b) a video upload to a web portal that enables the clinical assessment of behaviours in the family home. The findings were positive, finding there to be high agreement in terms of the diagnosis between remote methods and face to face methods and with high levels of satisfaction among the families and clinicians. This field is in the very early stages, and so only studies with small sample size were identified, but the findings suggest that there is potential for telehealth methods to improve assessment and diagnosis of autism used in conjunction with existing methods, especially for those with clear autism traits and adults with autism. Larger randomised controlled trials of this technology are warranted.

Keywords: assessment, autism spectrum disorder, diagnosis, telehealth

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Authors: Sukruthai Sapniwat, Falan Srisuriyachai

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Polymer Flooding is one of the most well-known methods in Enhanced Oil Recovery (EOR) technology which can be implemented after either primary or secondary recovery, resulting in favorable conditions for the displacement mechanism in order to lower the residual oil in the reservoir. Polymer substances can lower the mobility ratio of the whole process by increasing the viscosity of injected water. Therefore, polymer flooding can increase volumetric sweep efficiency, which leads to a better recovery factor. Moreover, polymer adsorption onto rock surface can help decrease reservoir permeability contrast with high heterogeneity. Due to the reduction of the absolute permeability, effective permeability to water, representing flow ability of the injected fluid, is also reduced. Once polymer is adsorbed onto rock surface, polymer molecule can be desorbed when different fluids are injected. This study is performed to evaluate the effects of the adsorption and desorption process of polymer solutions to yield benefits on the oil recovery mechanism. A reservoir model is constructed by reservoir simulation program called STAR® commercialized by the Computer Modeling Group (CMG). Various polymer concentrations, starting times of polymer flooding process and polymer injection rates were evaluated with selected values of polymer desorption degrees including 0, 25, 50, 75 and 100%. The higher the value, the more adsorbed polymer molecules to return back to flowing fluid. According to the results, polymer desorption lowers polymer consumption, especially at low concentrations. Furthermore, starting time of polymer flooding and injection rate affect the oil production. The results show that waterflooding followed by earlier polymer flooding can increase the oil recovery factor while the higher injection rate also enhances the recovery. Polymer concentration is related to polymer consumption due to the two main benefits of polymer flooding control described above. Therefore, polymer slug size should be optimized based on polymer concentration. Polymer desorption causes polymer re-employment that is previously adsorbed onto rock surface, resulting in an increase of sweep efficiency in the further period of polymer flooding process. Even though waterflooding supports polymer injectivity, water cut at the producer can prematurely terminate the oil production. The injection rate decreases polymer adsorption due to decreased retention time of polymer flooding process.

Keywords: enhanced oil recovery technology, polymer adsorption and desorption, polymer flooding, reservoir simulation

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Authors: Jocelyn Doucet, Jean-Philippe Laviolette, Ali Eslami

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The end-of-life management and recycling of polymer wastes remains a key environment issue in on-going efforts to increase resource efficiency and attaining GHG emission reduction targets. Half of all the plastics ever produced were made in the last 13 years, and only about 16% of that plastic waste is collected for recycling, while 25% is incinerated, 40% is landfilled, and 19% is unmanaged and leaks in the environment and waterways. In addition to the plastic collection issue, the UN recently published a report on chemicals in plastics, which adds another layer of challenge when integrating recycled content containing toxic products into new products. To tackle these important issues, innovative solutions are required. Chemical recycling of plastics provides new complementary alternatives to the current recycled plastic market by converting waste material into a high value chemical commodity that can be reintegrated in a variety of applications, making the total market size of the output – virgin-like, high value products - larger than the market size of the input – plastic waste. Access to high-quality feedstock also remains a major obstacle, primarily due to material contamination issues. Pyrowave approaches this challenge with its innovative nano-recycling technology, which purifies polymers at the molecular level, removing undesirable contaminants and restoring the resin to its virgin state without having to depolymerise it. This breakthrough approach expands the range of plastics that can be effectively recycled, including mixed plastics with various contaminants such as lead, inorganic pigments, and flame retardants. The technology allows yields below 100ppm, and purity can be adjusted to an infinitesimal level depending on the customer's specifications. The separation of the polymer and contaminants in Pyrowave's nano-recycling process offers the unique ability to customize the solution on targeted additives and contaminants to be removed based on the difference in molecular size. This precise control enables the attainment of a final polymer purity equivalent to virgin resin. The patented process involves dissolving the contaminated material using a specially formulated solvent, purifying the mixture at the molecular level, and subsequently extracting the solvent to yield a purified polymer resin that can directly be reintegrated in new products without further treatment. Notably, this technology offers simplicity, effectiveness, and flexibility while minimizing environmental impact and preserving valuable resources in the manufacturing circuit. Pyrowave has successfully applied this nano-recycling technology to decontaminate polymers and supply purified, high-quality recycled plastics to critical industries, including food-contact compliance. The technology is low-carbon, electrified, and provides 100% traceable resins with properties identical to those of virgin resins. Additionally, the issue of low recycling rates and the limited market for traditionally hard-to-recycle plastic waste has fueled the need for new complementary alternatives. Chemical recycling, such as Pyrowave's microwave depolymerization, presents a sustainable and efficient solution by converting plastic waste into high-value commodities. By employing microwave catalytic depolymerization, Pyrowave enables a truly circular economy of plastics, particularly in treating polystyrene waste to produce virgin-like styrene monomers. This revolutionary approach boasts low energy consumption, high yields, and a reduced carbon footprint. Pyrowave offers a portfolio of sustainable, low-carbon, electric solutions to give plastic waste a second life and paves the way to the new circular economy of plastics. Here, particularly for polystyrene, we show that styrene monomer yields from Pyrowave’s polystyrene microwave depolymerization reactor is 2,2 to 1,5 times higher than that of the thermal conventional pyrolysis. In addition, we provide a detailed understanding of the microwave assisted depolymerization via analyzing the effects of microwave power, pyrolysis time, microwave receptor and temperature on the styrene product yields. Furthermore, we investigate life cycle environmental impact assessment of microwave assisted pyrolysis of polystyrene in commercial-scale production. Finally, it is worth pointing out that Pyrowave is able to treat several tons of polystyrene to produce virgin styrene monomers and manage waste/contaminated polymeric materials as well in a truly circular economy.

Keywords: nanorecycling, nanomaterials, plastic recycling, depolymerization

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Authors: Chi-Chang Lin, Jian-Rong Wu, Jiun-Yan Chiu

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Myopia, ubiquitous symptom that is necessary to correct the eyesight by optical lens struggles many people for their daily life. Recent years, younger people raise interesting on using contact lens because of its convenience and aesthetics. In clinical, the risk of eye infections increases owing to the behavior of incorrectly using contact lens unsupervised cleaning which raising the infection risk of cornea, named ocular keratitis. In order to overcome the identification needs, new detection or analysis method with rapid and more accurate identification for clinical microorganism is importantly needed. In our study, we take advantage of Raman spectroscopy having unique fingerprint for different functional groups as the distinct and fast examination tool on microorganism. As we know, Raman scatting signals are normally too weak for the detection, especially in biological field. Here, we applied special SERS enhancement substrates to generate higher Raman signals. SERS filter we designed in this article that prepared by deposition of silver nanoparticles directly onto cellulose filter surface and suspension nanoparticles - gold nanostars (AuNSs) also be introduced together to achieve better enhancement for lower concentration analyte (i.e., various bacteria). Research targets also focusing on studying the shape effect of synthetic AuNSs, needle-like surface morphology may possible creates more hot-spot for getting higher SERS enhance ability. We utilized new designed SERS technology to distinguish the bacteria from ocular keratitis under strain level, and specific Raman and SERS fingerprint were grouped under pattern recognition process. We reported a new method combined different SERS substrates can be applied for clinical microorganism detection under strain level with simple, rapid preparation and low cost. Our presenting SERS technology not only shows the great potential for clinical bacteria detection but also can be used for environmental pollution and food safety analysis.

Keywords: bacteria, gold nanostars, Raman spectroscopy surface-enhanced Raman scattering filter

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Authors: Denise Levy, Anna Lucia C. H. Villavicencio

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In a rapid-changing world, science teachers face considerable challenges. In addition to the basic curriculum, there must be included several transversal themes, which demand creative and innovative strategies to be arranged and integrated to traditional disciplines. In Brazil, nuclear science is still a controversial theme, and teachers themselves seem to be unaware of the issue, most often perpetuating prejudice, errors and misconceptions. This article presents the authors’ experience in the development of an interdisciplinary pedagogical proposal to include nuclear science in the basic curriculum, in a transversal and integrating way. The methodology applied was based on the analysis of several normative documents that define the requirements of essential learning, competences and skills of basic education for all schools in Brazil. The didactic materials and resources were developed according to the best practices to improve learning processes privileging constructivist educational techniques, with emphasis on active learning process, collaborative learning and learning through research. The material consists of an illustrated book for students, a book for teachers and a manual with activities that can articulate nuclear science to different disciplines: Portuguese, mathematics, science, art, English, history and geography. The content counts on high scientific rigor and articulate nuclear technology with topics of interest to society in the most diverse spheres, such as food supply, public health, food safety and foreign trade. Moreover, this pedagogical proposal takes advantage of the potential value of digital technologies, implementing QR codes that excite and challenge students of all ages, improving interaction and engagement. The expected results include the education of the educators for nuclear science communication in a transversal and integrating way, demystifying nuclear technology in a contextualized and significant approach. It is expected that the interdisciplinary pedagogical proposal contributes to improving attitudes towards knowledge construction, privileging reconstructive questioning, fostering a culture of systematic curiosity and encouraging critical thinking skills.

Keywords: science education, interdisciplinary learning, nuclear science, scientific literacy

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Authors: K. Raghu, Sree Harsha

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The interrelationship between the technology and architecture of tall buildings is investigated from the emergence of tall buildings in late 19th century to the present. In the late 19th century early designs of tall buildings recognized the effectiveness of diagonal bracing members in resisting lateral forces. Most of the structural systems deployed for early tall buildings were steel frames with diagonal bracings of various configurations such as X, K, and eccentric. Though the historical research a filtering concept is developed original and remedial technology- through which one can clearly understand inter-relationship between the technical evolution and architectural esthetic and further stylistic transition buildings. Diagonalized grid structures – “diagrids” - have emerged as one of the most innovative and adaptable approaches to structuring buildings in this millennium. Variations of the diagrid system have evolved to the point of making its use non-exclusive to the tall building. Diagrid construction is also to be found in a range of innovative mid-rise steel projects. Contemporary design practice of tall buildings is reviewed and design guidelines are provided for new design trends. Investigated in depths are the behavioral characteristics and design methodology for diagrids structures, which emerge as a new direction in the design of tall buildings with their powerful structural rationale and symbolic architectural expression. Moreover, new technologies for tall building structures and facades are developed for performance enhancement through design integration, and their architectural potentials are explored. By considering the above data the analysis and design of 40-100 storey diagrids steel buildings is carried out using E-TABS software with diagrids of various angle to be found for entire building which will be helpful to reduce the steel requirement for the structure. The present project will have to undertake wind analysis, seismic analysis for lateral loads acting on the structure due to wind loads, earthquake loads, gravity loads. All structural members are designed as per IS 800-2007 considering all load combination. Comparison of results in terms of time period, top storey displacement and inter-storey drift to be carried out. The secondary effect like temperature variations are not considered in the design assuming small variation.

Keywords: diagrid, bracings, structural, building

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Authors: Soheir Mohamed Hegazy

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Culture sustainability, which reflects a deep respect for people and history, is a cause of concern in contemporary architecture. Adopting ultramodern architecture styles was initiated in the 20th century by a plurality of states worldwide. Only a few countries, including Oman, realized that fashionable architectural designs ignore cultural values, identity, the context of its environment, economic perspective, and social performance. Stirring the Sultanate of Oman from being a listless and closed community to a modern country started in the year 1970. Despite unprecedented development in all aspects of Omani people's life, the leadership and the public had the capability to adjust to the changing global challenges without compromising social values and identity. This research provides a close analysis of one of the recent examples of contemporary vernacular architecture in the Sultanate of Oman, as a case study, Oman International Airport. The said airport gained an international appreciation for its Omani-themed architecture, distinguished traveler experience, and advanced technology. Accordingly, it was selected by the World Travel Awards as the Best Tourism Development Project in the Middle East only four weeks afterward after starting its operation. This paper aims to transfer this successful design approach of integrating the latest trends in technology, systems, eco-friendly aspects, and materials with the traditional Omani architectural features, which reflects symbiotic harmony of the community, individuals, and environment to other countries, designers, researchers, and students. In addition, the paper aims to encourage architects and teachers to take responsibility for valorizing built heritage as a source of inspiration for modern architecture, which could be considered as an added value. The work depends on reviewing the relevant literature, a case study, interviews with two architects who were involved in the project’s site work, and one current high-ranking employee in the airport besides data analysis and conclusion.

Keywords: contemporary vernacular architecture, culture sustainability, Oman international airport, current Omani architecture type

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Authors: Nizar Chaira

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Dates, main products of the oases, due to their therapeutic interests, are considered highly nutritious fruit. Several studies on the valuation biotechnology and technology of dates are made, and several products are already prepared. Isolation of the yeast Saccharomyces cerevisiae, naturally presents in a scrap of date, optimization of growth in the medium based on date syrup and production biomass can potentially expand the range of secondary products of dates. To this end, this paper tries to study the suitability for processing dates technology and biotechnology to use the date pulp as a carbon source for biological transformation. Two strains of Saccharomyces cerevisiae isolated from date syrup (S1, S2) and a commercial strain have used for this study. After optimization of culture conditions, production in a fermenter on two different media (date syrup and beet molasses) was performed. This is followed by studying the kinetics of growth, protein production and consumption of sugars in crops strain 1, 2 and the commercial strain and on both media. The results obtained showed that a concentration of 2% sugar, 2.5 g/l yeast extract, pH 4.5 and a temperature between 25 and 35°C are the optimal conditions for cultivation in a bioreactor. The exponential phase of the specific growth rate of a strain on both media showed that it is about 0.3625 h-1 for the production of a medium based on date syrup and 0.3521 h-1 on beet molasses with a generation time equal to 1.912 h and on the medium based on date syrup, yeast consumes preferentially the reducing sugars. For the production of protein, we showed that this latter presents an exponential phase when the medium starts to run out of reducing sugars. For strain 2, the specific growth rate is about 0.261h-1 for the production on a medium based on date syrup and 0207 h-1 on beet molasses and the base medium syrup date of the yeast consumes preferentially reducing sugars. For the invertase and other metabolits, these increases rapidly after exhaustion of reducing sugars. The comparison of productivity between the three strains on the medium based on date syrup showed that the maximum value is obtained with the second strain: p = 1072 g/l/h as it is about of 0923 g/l/h for strain 1 and 0644 g/l/h for the commercial strain. Thus, isolates of date syrup are more competitive than the commercial strain and can give the same performance in a shorter time with energy gain.

Keywords: date palm, fermentation, molasses, Saccharomyces, syrup

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Authors: Delphine Morales, Florian Lombart, Agathe Truchot, Pauline Maire, Pascale Vigneron, Antoine Galmiche, Catherine Lok, Muriel Vayssade

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Targeted therapy molecules are used as a first-line treatment for metastatic melanoma with B-Raf mutation. Nevertheless, these molecules can cause side effects to patients and are efficient on 50 to 60 % of them. Indeed, melanoma cell sensitivity to targeted therapy molecules is dependent on tumor microenvironment (cell-cell and cell-extracellular matrix interactions). To better unravel factors modulating cell sensitivity to B-Raf inhibitor, we have developed and compared several melanoma models: from metastatic melanoma cells cultured as monolayer (2D) to a co-culture in a 3D dermal equivalent. Cell response was studied in different melanoma cell lines such as SK-MEL-28 (mutant B-Raf (V600E), sensitive to Vemurafenib), SK-MEL-3 (mutant B-Raf (V600E), resistant to Vemurafenib) and a primary culture of dermal human fibroblasts (HDFn). Assays have initially been performed in a monolayer cell culture (2D), then a second time on a 3D dermal equivalent (dermal human fibroblasts embedded in a collagen gel). All cell lines were treated with Vemurafenib (a B-Raf inhibitor) for 48 hours at various concentrations. Cell sensitivity to treatment was assessed under various aspects: Cell proliferation (cell counting, EdU incorporation, MTS assay), MAPK signaling pathway analysis (Western-Blotting), Apoptosis (TUNEL), Cytokine release (IL-6, IL-1α, HGF, TGF-β, TNF-α) upon Vemurafenib treatment (ELISA) and histology for 3D models. In 2D configuration, the inhibitory effect of Vemurafenib on cell proliferation was confirmed on SK-MEL-28 cells (IC50=0.5 µM), and not on the SK-MEL-3 cell line. No apoptotic signal was detected in SK-MEL-28-treated cells, suggesting a cytostatic effect of the Vemurafenib rather than a cytotoxic one. The inhibition of SK-MEL-28 cell proliferation upon treatment was correlated with a strong expression decrease of phosphorylated proteins involved in the MAPK pathway (ERK, MEK, and AKT/PKB). Vemurafenib (from 5 µM to 10 µM) also slowed down HDFn proliferation, whatever cell culture configuration (monolayer or 3D dermal equivalent). SK-MEL-28 cells cultured in the dermal equivalent were still sensitive to high Vemurafenib concentrations. To better characterize all cell population impacts (melanoma cells, dermal fibroblasts) on Vemurafenib efficacy, cytokine release is being studied in 2D and 3D models. We have successfully developed and validated a relevant 3D model, mimicking cutaneous metastatic melanoma and tumor microenvironment. This 3D melanoma model will become more complex by adding a third cell population, keratinocytes, allowing us to characterize the epidermis influence on the melanoma cell sensitivity to Vemurafenib. In the long run, the establishment of more relevant 3D melanoma models with patients’ cells might be useful for personalized therapy development. The authors would like to thank the Picardie region and the European Regional Development Fund (ERDF) 2014/2020 for the funding of this work and Oise committee of "La ligue contre le cancer".

Keywords: 3D human skin model, melanoma, tissue engineering, vemurafenib efficiency

Procedia PDF Downloads 290

Authors: Ipsita Biswas, Arnab Sarkar, Ashikur Rahaman, Gopeswar Mukherjee, Subhrangsu Chatterjee, Shamee Bhattacharjee, Deba Prasad Mandal

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One of the major reasons for the high mortality rate of lung cancer is the substantial delays in disease detection at late metastatic stages. It is of utmost importance to understand the detailed molecular signaling and detect the molecular markers that can be used for the early diagnosis of cancer. Several studies explored the emerging roles of long noncoding RNAs (lncRNAs) in various cancers as well as lung cancer. A long non-coding RNA LINC00273 was recently discovered to promote cancer cell migration and invasion, and its positive correlation with the pathological stages of metastasis may prove it to be a potential target for inhibiting cancer cell metastasis. Comparing real-time expression of LINC00273 in various human clinical cancer tissue samples with normal tissue samples revealed significantly higher expression in cancer tissues. This long intergenic noncoding RNA was found to be highly expressed in human liver tumor-initiating cells, human gastric adenocarcinoma AGS cell line, as well as human non-small cell lung cancer A549 cell line. SiRNA and shRNA-induced knockdown of LINC00273 in both in vitro and in vivo nude mice significantly subsided AGS and A549 cancer cell migration and invasion. LINC00273 knockdown also reduced TGF-β induced SNAIL, SLUG, VIMENTIN, ZEB1 expression, and metastasis in A549 cells. Plenty of reports have suggested the role of microRNAs of the miR200 family in reversing epithelial to mesenchymal transition (EMT) by inhibiting ZEB transcription factors. In this study, hsa-miR-200a-3p was predicted via IntaRNA-Freiburg RNA tools to be a potential target of LINC00273 with a negative free binding energy of −8.793 kcal/mol, and this interaction was verified as a confirmed target of LINC00273 by RNA pulldown, real-time PCR and luciferase assay. Mechanistically, LINC00273 accelerated TGF-β induced EMT by sponging hsa-miR-200a-3p which in turn liberated ZEB1 and promoted prometastatic functions in A549 cells in vitro as verified by real-time PCR and western blotting. The similar expression patterns of these EMT regulatory pathway molecules, viz. LINC00273, hsa-miR-200a-3p, ZEB1 and TGF-β, were also detected in various clinical samples like breast cancer tissues, oral cancer tissues, lung cancer tissues, etc. Overall, this LINC00273 mediated EMT regulatory signaling can serve as a potential therapeutic target for the prevention of lung cancer metastasis.

Keywords: epithelial to mesenchymal transition, long noncoding RNA, microRNA, non-small-cell lung carcinoma

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Authors: Jiri Barta, Jiří F. Urbanek

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This paper deals with a protection of the national and European infrastructure. It is issue nowadays. The paper deals with the perspectives and possibilities of "smart solutions" to critical infrastructure protection. The research project deals with computers aided technologies are used from the perspective of new, better protection of selected infrastructure objects. Protection is focused on communication and information channels. These communication and information channels are very important for the functioning of the system of protection of critical infrastructure elements.

Keywords: interoperability, communication systems, controlling proces, critical infrastructure, crisis workplaces, continuity

Procedia PDF Downloads 283

Authors: Z. Karahaliloğlu, E. Yalçın, M. Demirbilek, E.B. Denkbaş

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Critical-sized bone defects caused by trauma, bone diseases, prosthetic implant revision or tumor excision cannot be repaired by physiological regenerative processes. Current orthopedic applications for critical-sized bone defects are to use autologous bone grafts, bone allografts, or synthetic graft materials. However, these strategies are unable to solve completely the problem, and motivate the development of novel effective biological scaffolds for tissue engineering applications and regenerative medicine applications. In particular, scaffolds combined with a variety of bio-agents as fundamental tools emerge to provide the regeneration of damaged bone tissues due to their ability to promote cell growth and function. In this study, a magnetic silk fibroin (SF) hydrogel scaffold was prepared by electrogelation process of the concentrated Bombxy mori silk fibroin (8 %wt) aqueous solution. For enhancement of osteoblast-like cells (SaOS-2) growth and adhesion, basal fibroblast growth factor (bFGF) were conjugated physically to the HSA-coated magnetic nanoparticles (Fe3O4) and magnetic SF e-gel scaffolds were prepared by incorporation of Fe3O4, HSA (human serum albumin)=Fe3O4 and HSA=Fe3O4-bFGF nanoparticles. HSA=Fe3O4, HSA=Fe3O4-bFGF loaded and bare SF e-gels scaffolds were characterized using scanning electron microscopy (SEM.) For cell studies, human osteoblast-like cell line (SaOS-2) was used and an MTT assay was used to assess the cytotoxicity of magnetic silk fibroin e-gel scaffolds and cell density on these surfaces. For the evaluation osteogenic activation, ALP (alkaline phosphatase), the amount of mineralized calcium, total protein and collagen were studied. Fe3O4 nanoparticles were successfully synthesized and bFGF was conjugated to HSA=Fe3O4 nanoparticles with %97.5 of binding yield which has a particle size of 71.52±2.3 nm. Electron microscopy images of the prepared HSA and bFGF incorporated SF e-gel scaffolds showed a 3D porous morphology. In terms of water uptake results, bFGF conjugated HSA=Fe3O4 nanoparticles has the best water absorbability behavior among all groups. In the in-vitro cell culture studies realized using SaOS-2 cell line, the coating of Fe3O4 nanoparticles surface with a protein enhance the cell viability and HSA coating and bFGF conjugation, the both have an inductive effect in the cell proliferation. One of the markers of bone formation and osteoblast differentiation, according to the ALP activity and total protein results, HSA=Fe3O4-bFGF loaded SF e-gels had significantly enhanced ALP activity. Osteoblast cultured HSA=Fe3O4-bFGF loaded SF e-gels deposited more calcium compared with SF e-gel. The proposed magnetic scaffolds seem to be promising for bone tissue regeneration and used in future work for various applications.

Keywords: basic fibroblast growth factor (bFGF), e-gel, iron oxide nanoparticles, silk fibroin

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Authors: Malik Muhammad Arslan, Muneeb Ullah, Dai Shihan, Daniyal Haider, Xiaodong Yang

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Chest X-rays are instrumental in the detection and monitoring of a wide array of diseases, including viral infections such as COVID-19, tuberculosis, pneumonia, lung cancer, and various cardiac and pulmonary conditions. To enhance the accuracy of diagnosis, artificial intelligence (AI) algorithms, particularly deep learning models like Convolutional Neural Networks (CNNs), are employed. However, these deep learning models demand a substantial and varied dataset to attain optimal precision. Generative Adversarial Networks (GANs) can be employed to create new data, thereby supplementing the existing dataset and enhancing the accuracy of deep learning models. Nevertheless, GANs have their limitations, such as issues related to stability, convergence, and the ability to distinguish between authentic and fabricated data. In order to overcome these challenges and advance the detection and classification of CXR normal and abnormal images, this study introduces a distinctive technique known as DCWGAN (Diverse Conditional Wasserstein GAN) for generating synthetic chest X-ray (CXR) images. The study evaluates the effectiveness of this Idiosyncratic DCWGAN technique using the ResNet50 model and compares its results with those obtained using the traditional GAN approach. The findings reveal that the ResNet50 model trained on the DCWGAN-generated dataset outperformed the model trained on the classic GAN-generated dataset. Specifically, the ResNet50 model utilizing DCWGAN synthetic images achieved impressive performance metrics with an accuracy of 0.961, precision of 0.955, recall of 0.970, and F1-Measure of 0.963. These results indicate the promising potential for the early detection of diseases in CXR images using this Inimitable approach.

Keywords: CNN, classification, deep learning, GAN, Resnet50

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Authors: Larasati Zuhro, Ida F Priyanto

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Background: In March 2020, Indonesia started to be affected by Covid-19, and the number of victims increased slowly but surely until finally, the highest number of victims reached the highest—about 50,000 persons—for the daily cases in the middle of 2021. Like other institutions, schools and universities were suddenly closed in March 2020, and students had to change their ways of studying from face-to-face to online. This sudden changed affected students and faculty, including LIS students and faculty because they never experienced online classes in Indonesia due to the previous regulation that academic and school activities were all conducted onsite. For almost two years, school and academic activities were held online. This indeed has affected the way students learned and faculty delivered their courses. This raises the question of whether students are now ready for their new learning activities due to the covid-19 disruption. Objectives: this study was conducted to find out the impact of covid-19 pandemic on the LIS learning process and the effectiveness of online classes for students of LIS in Indonesia. Methodology: This was qualitative research conducted among LIS students at UIN Sunan Kalijaga, Yogyakarta, Indonesia. The population are students who were studying for masters’program during covid-19 pandemic. Results: The study showed that students were ready with the online classes because they are familiar with the technology. However, the Internet and technology infrastructure do not always support the process of learning. Students mention slow WIFI is one factor that causes them not being able to study optimally. They usually compensate themselves by visiting a public library, a café, or any other places to get WIFI network. Noises come from the people surrounding them while they are studying online.Some students could not concentrate well when attending the online classes as they studied at home, and their families sometimes talk to other family members, or they asked the students while they are attending the online classes. The noise also came when they studied in a café. Another issue is that the classes were held in shorter time than that in the face-to-face. Students said they still enjoyed the onsite classes instead of online, although they do not mind to have hybrid model of learning. Conclusion: Pandemic of Covid-19 has changed the way students of LIS in Indonesia learn. They have experienced a process of migrating the way they learn from onsite to online. They also adapted their learning with the condition of internet access speed, infrastructure, and the environment. They expect to have hybrid classes in the future.

Keywords: learning, LIS students, pandemic, covid-19

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Authors: Samia Moulebhar, Chahrazed Bendenia, Souhila Bendenia, Hanaa Merad-dib, Sarra Merabet, Sid Ahmed Khantar, Baghdad Hadri

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The photovoltaic solar field is today experiencing exponential advancement with the exploitation of new technological sectors of nanoparticles, namely the field of solar cells based on organic polymer materials. These cells are flexible, easy to process and low cost. This work includes a presentation of the conjugated polymer materials used in the design of photovoltaic technology devices while determining their properties and then the models used for the modeling of thin film photovoltaic cells heterojunction.

Keywords: photovoltaic, cells, nanoparticles, organic

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Authors: M. Tsebia, H. Bentarzi

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In the inter-connected power systems, a phenomenon called inter-area oscillation may be caused by several defects. In this paper, a study of the Maghreb countries inter-area power networks oscillation has been investigated. The inter-area oscillation monitoring can be enhanced by integrating Phasor Measurement Unit (PMU) technology installed in different places. The data provided by PMU and recorded by PDC will be used for the monitoring, analysis, and control purposes. The proposed approach has been validated by simulation using MATLAB/Simulink.

Keywords: PMU, inter-area oscillation, Maghrebian power system, Simulink

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Authors: Danutė Jablonskytė-Raščė, Vidas Damanauskas

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The wheat (Triticum) genus is divided into many species, of which only two are widely distributed in the world - common wheat (Triticum aestivum L.) and durum wheat (Triticum durum Desf.). Common (soft) wheat is the most common type of wheat in the world and the most suitable for the harsh climate of Lithuania, but the grains have lower protein content and poorer nutritional properties. Durum wheat is characterized by a high protein content of the grain, but it is a crop of warmer climates grown in southern countries, Italy, Spain, the United States, Egypt, etc. Today's important issue is food, its resources and quality. The research focuses on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the warming climate conditions. Climatic conditions change the distribution of fungi and their hosts. Plants that have grown in our climate for many years have adapted to the use of fungicides, so the aim is to study cereal varieties grown in warmer climates and compare them with our country's varieties, studying their nutritional value and the spread of fungal diseases. The field experiments of different varieties of wheat were conducted at Joniškėlis Experimental Station of the Lithuanian Research Centre for Agriculture and Forestry in 2023. The soil of the experimental site was Endocalcari-Endohypogleyic Cambisol (CMg-n-w-can). The research was designed to identify the resistance to leaf diseases and the nutritional value of various wheat varieties. This research aims to focus on healthier food grown in our conditions, the quality of which recently depends a lot not only on the cultivation technology but also on the conditions of the warming climate. The study found that hot and humid summer weather led to the spread of foliar diseases in wheat. Tan spot (Pyrenophora tritici-repentis) is mostly spread in wheat crops. This disease had an average prevalence of 86.90%. The wheat crop was sparse, so this year was unfavorable for the spread of powdery mildew (Blumeria graminis). Dry weather prevailed during the period of flowering of cereals, which prevented the spread of ear diseases. Examining the qualitative indicators of grain, it was found that durum wheat had the best parameters.

Keywords: varieties, wheat, leaf disease, grain quality

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Authors: Khaldon Lweesy, Feryal Alskafi, Rabaa Hammad, Shaker Khanfar, Yara Alsukhni

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Electronic noses are devices designed to emulate the humane sense of smell by characterizing and differentiating odor profiles. In this study, we build a low-cost e-nose using an array module containing four different types of metal oxide semiconductor gas sensors. We used this system to create a profile for a meat specimen over three days. Then using a pattern recognition software, we correlated the odor of the specimen to its age. It is a simple, fast detection method that is both non-expensive and non-destructive. The results support the usage of this technology in food control management.

Keywords: e-nose, low cost, odor detection, food safety

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Authors: Anas Hallak, Latifa Seblini, Juergen Wilde

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In mechatronics or sensor technology, plastic housings are used to protect sensitive components from harmful environmental influences, such as moisture, media, or reactive substances. Connections, preferably in the form of metallic lead-frame structures, through the housing wall are required for their electrical supply or control. In this system, an insufficient connection between the plastic component, e.g., Polyamide66, and the metal surface, e.g., copper, due to the incompatibility is dominating. As a result, leakage paths can occur along with the plastic-metal interface. Since adhesive bonding has been established as one of the most important joining processes and its use has expanded significantly, driven by the development of improved high-performance adhesives and bonding techniques, this technology has been involved in metal-plastic hybrid structures. In this study, an epoxy bonding agent from DELO (DUALBOND LT2266) has been used to improve the mechanical and chemical binding between the metal and the polymer. It is an adhesion promoter with two reaction stages. In these, the first stage provides fixation to the lead frame directly after the coating step, which can be done by UV-Exposure for a few seconds. In the second stage, the material will be thermally hardened during injection molding. To analyze the two reaction stages of the primer, dynamic DSC experiments were carried out and correlated with Fourier-transform infrared spectroscopy measurements. Furthermore, the number of crosslinking bonds formed in the system in each reaction stage has also been estimated by a rheological characterization. Those investigations have been performed with different times of UV exposure: 12, 96 s and in an industrial preferred temperature range from -20 to 175°C. The shear viscosity values of primer have been measured as a function of temperature and exposure times. For further interpretation, the storage modulus values have been calculated, and the so-called Booij–Palmen plot has been sketched. The next approach in this study is the self-healing mechanisms in the hydride system in which the primer should flow into micro-damage such as interface, cracks, inhibit them from growing, and close them. The ability of the primer to flow in and penetrate defined capillaries made in Ultramid was investigated. Holes with a diameter of 0.3 mm were produced in injection-molded A3EG7 plates with 4 mm thickness. A copper substrate coated with the DUALBOND was placed on the A3EG7 plate and pressed with a certain force. Metallographic analyses were carried out to verify the filling grade, which showed an almost 95% filling ratio of the capillaries. Finally, to estimate the self-healing mechanism in metal-plastic hybrid systems, characterizations have been done on a simple geometry with a metal inlay developed by the Institute of Polymer Technology in Friedrich-Alexander-University. The specimens have been modified with tungsten wire which was to be pulled out after the injection molding to create a micro-hole in the specimen at the interface between the primer and the polymer. The capability of the primer to heal those micro-cracks upon heating, pressing, and thermal aging has been characterized through metallographic analyses.

Keywords: hybrid structures, self-healing, thermoplastic housing, adhesive

Procedia PDF Downloads 177

Authors: Jun Seung Woo, Shin Dong Ho

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The Image Captioning models with Attention technology have developed significantly compared to previous models, but it is still unsatisfactory in recognizing images. We perform an extensive search over seven interesting Convolutional Neural Networks(CNN) architectures to analyze the behavior of different models for image captioning. We compared seven different CNN Architectures, according to batch size, using on public benchmarks: MS-COCO datasets. In our experimental results, DenseNet and InceptionV3 got about 14% loss and about 160sec training time per epoch. It was the most satisfactory result among the seven CNN architectures after training 50 epochs on GPU.

Keywords: deep learning, image captioning, CNN architectures, densenet, inceptionV3

Procedia PDF Downloads 114

Authors: I. Upadhyaya, K. Arsi, A. M. Donoghue, C. N. Coon, M. Schlumbohm, M. N. Riaz, M. B. Farnell, A. Upadhyay, A. J. Davis, D. J. Donoghue

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Methionine, a sulfur containing amino acid, is essential for healthy poultry production. Synthetic methionine is commonly used as a supplement in conventional poultry. However, for organic poultry, a natural, cost effective source of methionine that can replace synthetic methionine is unavailable. Invasive Asian carp (AC) are a potential natural methionine source; however, there is no proven technology to utilize this fish methionine. Commercially available rendering is environmentally challenging due to the offensive smell produced during production. We explored extrusion technology as a potential cost effective alternative to fish rendering. We also determined the amino acid composition, digestible amino acids and total metabolizable energy (TMEn) for the extruded AC fish meal. Dry extrusion of AC was carried out by mixing the fish with soybean meal (SBM) in a 1:1 proportion to reduce high moisture in the fishmeal using an Insta Pro Jr. dry extruder followed by drying and grinding of the product. To determine the digestible amino acids and TMEn of the extruded product, a colony of cecectomized Bovans White Roosters was used. Adult roosters (48 weeks of age) were fasted for 30 h and tube fed 35 grams of 3 treatments: (1) extruded AC fish meal, (2) SBM and (3) corn. Excreta from each individual bird was collected for the next 48 h. An additional 10 unfed roosters served as endogenous controls. The gross energy and protein content of the feces from the treatments were determined to calculate the TMEn. Fecal samples and treatment feeds were analyzed for amino acid content and percent digestible amino acid. Results from the analysis suggested that addition of Asian carp increased the methionine content of SBM from 0.63 to 0.83%. Also, the digestibility of amino acid and the TMEn values were greater for the AC meal with SBM than SBM alone. The dry extruded AC meal analysis is indicative that the product can replace SBM alone and enhance natural methionine in a standard poultry ration. The results from feed formulation using different concentrations of the AC fish meal depict a potential diet which can supplement the required methionine content in organic poultry production.

Keywords: Asian carp, extrusion, natural methionine, organic poultry

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Authors: Kai-Shiang Chang, Shiao-Shing Chen, Saikat Sinha Ray, Hung-Te Hsu

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In recent years, membrane bioreactors (MBR) have been widely utilized as it can effectively replace conventional activated sludge process (CAS). Membrane bioreactor (MBR) is found to be more effective technology compared to other conventional activated sludge process and advanced membrane separation technique. Additionally, as far as the MBR is concerned, it is having excellent control of sludge retention time (SRT) and hydraulic retention time (HRT) and conducive to the retention of high concentration of sludge biomass. The membrane bioreactor (MBR) can effectively reduce footprint in terms of area and omit the secondary processing procedures in the conventional activated sludge process (CAS). Currently, as per the membrane technology, the ceramic membrane is found to have highly strong anti-acid-base properties, and it is more suitable than polymeric membrane while using for backwash and chemical cleaning. This study is based upon the treatment of Cutting Fluid wastewater, as the Cutting Fluid is widely used in the cutting equipment. However, the Cutting Fluid wastewater is very difficult to treat. In this study, the ceramic membrane was used and combine with of MBR system to treat the Cutting Fluid wastewater. In this present study, different kind of chemical coagulants have been utilized for pretreatment purpose in order to get the supernatant and simultaneously this wastewater (supernatant) was treated by MBR process. Nevertheless, ceramic membrane has three advantages such as high mechanical strength, drug resistance and reuse. During the experiment, the backwash technique was used for every interval of 10 minutes in order to avoid fouling of the membrane. In this study, during pretreatment the Chemical Oxygen Demand (COD) removal efficiency was found to be 71-86% and oil removal efficiency was analyzed to be 83-92%. This pretreatment study suggests that it is quiet effective methodology to reduce COD and oil concentration. Finally, In the MBR system when the HRT is more than 7.5 hour, the COD removal efficiency was found to be 87-93% and could achieve 100% oil removal efficiency. Coagulation test series were seen in Refs coagulants for the treatment of wastewater containing cutting oil with better oil and COD removal efficiency. The results also showed that the oil removal efficiency in the MBR system could reduce the oil content to less than 1 mg / L when the oil quality was 126 mg / L. Therefore, in this paper, the performance of membrane bioreactor by utilizing ceramic membrane has been demonstrated for treatment of Cutting Fluid wastewater.

Keywords: membrane bioreactor, cutting fluid, oil, chemical oxygen demand

Procedia PDF Downloads 299

Authors: Asim Qayyum, David Smith

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The use of information tools is a common activity for students of any educational stage when they encounter online learning activities. Finding the relevant information for particular learning tasks is the topic of this paper as it investigates the use of information tools for a group of student participants. The paper describes and discusses the results with particular implications for use in higher education, and the findings suggest that improvement in assessment design and subsequent student learning may be achieved by structuring the purposefulness of information tools usage and online reading behaviors of university students.

Keywords: information tools, assessment, online learning, student assessment experience

Procedia PDF Downloads 540

Authors: Lamzira Ebralidze, Aleksandre Tsertsvadze, Dali Berashvili, Aliosha Bakuridze

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Nowadays, there is an increasing demand for biologically active substances from vegetable, animal, and mineral resources. In terms of the use of natural compounds, pharmaceutical, cosmetic, and nutrition industry has big interest. The biggest drawback of conventional extraction methods is the need to use a large volume of organic extragents. The removal of the organic solvent is a multi-stage process. And their absolute removal cannot be achieved, and they still appear in the final product as impurities. A large amount of waste containing organic solvent damages not only human health but also has the harmful effects of the environment. Accordingly, researchers are focused on improving the extraction methods, which aims to minimize the use of organic solvents and energy sources, using alternate solvents and renewable raw materials. In this context, green extraction principles were formed. Green Extraction is a need of today’s environment. Green Extraction is the concept, and it totally corresponds to the challenges of the 21st century. The extraction of biologically active compounds based on green extraction principles is vital from the view of preservation and maintaining biodiversity. Novel technologies of green extraction are known, such as "cold methods" because during the extraction process, the temperature is relatively lower, and it doesn’t have a negative impact on the stability of plant compounds. Novel technologies provide great opportunities to reduce or replace the use of organic toxic solvents, the efficiency of the process, enhance excretion yield, and improve the quality of the final product. The objective of the research is the development of green technologies of flavonoids containing preparations. Methodology: At the first stage of the research, flavonoids containing preparations (Tincture Herba Leonuri, flamine, rutine) were prepared based on conventional extraction methods: maceration, bismaceration, percolation, repercolation. At the same time, the same preparations were prepared based on green technologies, microwave-assisted, UV extraction methods. Product quality characteristics were evaluated by pharmacopeia methods. At the next stage of the research technological - economic characteristics and cost efficiency of products prepared based on conventional and novel technologies were determined. For the extraction of flavonoids, water is used as extragent. Surface-active substances are used as co-solvent in order to reduce surface tension, which significantly increases the solubility of polyphenols in water. Different concentrations of water-glycerol mixture, cyclodextrin, ionic solvent were used for the extraction process. In vitro antioxidant activity will be studied by the spectrophotometric method, using DPPH (2,2-diphenyl-1- picrylhydrazyl) as an antioxidant assay. The advantage of green extraction methods is also the possibility of obtaining higher yield in case of low temperature, limitation extraction process of undesirable compounds. That is especially important for the extraction of thermosensitive compounds and maintaining their stability.

Keywords: extraction, green technologies, natural resources, flavonoids

Procedia PDF Downloads 116

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

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

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

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Authors: Jong-Bae Lee, Seongsoo Lee

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Local interconnect network (LIN) is a communication protocol that combines sensors, actuators, and processors to a functional module in automotive applications. In this paper, a LIN ver. 2.2A controller was designed in Verilog hardware description language (Verilog HDL) and implemented in field-programmable gate array (FPGA). Its operation was verified by making full-scale LIN network with the presented FPGA-implemented LIN controller, commercial LIN transceivers, and commercial processors. When described in Verilog HDL and synthesized in 0.18 μm technology, its gate size was about 2,300 gates.

Keywords: local interconnect network, controller, transceiver, processor

Procedia PDF Downloads 270

Authors: M. R. Babaei, S. M. Hosseini, S. Rahmani, L. Moradi

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Today, the quality of education and training systems and the effectiveness of the education systems of most concern to stakeholders and decision-makers of our country's development in each country. In Iran this is a double issue of concern to numerous reasons; So that governments, over the past decade have hardly even paid the running costs of education. ICT is claiming it has the power to change the structure of a program for training, reduce costs and increase quality, and do education systems and products consistent with the needs of the community and take steps to practice education. Own of the areas that the introduction of information technology has fundamentally changed is the field of education. The aim of this research is process reengineering management in schools simultaneously has been using field studies to collect data in the form of interviews and a questionnaire survey. The statistical community of this research has been the country of Iran and smart schools under the education. Sampling was targeted. The data collection tool was a questionnaire composed of two parts. The questionnaire consists of 36 questions that each question designates one of effective factors on the management of smart schools. Also each question consists of two parts. The first part designates the operating position in the management process, which represents the domain's belonging to the management agent (planning, organizing, leading, controlling). According to the classification of Dabryn and in second part the factors affect the process of managing the smart schools were examined, that Likert scale is used to classify. Questions the validity of the group of experts and prominent university professors in the fields of information technology, management and reengineering of approved and Cronbach's alpha reliability and also with the use of the formula is evaluated and approved. To analyse the data, descriptive and inferential statistics were used to analyse the factors contributing to the rating of (Linkert scale) descriptive statistics (frequency table data, mean, median, mode) was used. To analyse the data using analysis of variance and nonparametric tests and Friedman test, the assumption was evaluated. The research conclusions show that the factors influencing the management process re-engineering smart schools in school performance is affected.

Keywords: re-engineering, management process, smart school, Iran's school

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Authors: Mihretu Erjabo

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Halaba special district is characterized by drought, soil erosion, high population pressure, poor livestock production, lack of feed for livestock, very deep water table, very low productivity of crops and food insufficiency. In order to address these problems, the woreda agricultural development office along with other management practices such as soil physical conservation measures agroforestry was introduced decades ago as a means to alleviate the problem. However, the level of agroforestry adoption remains low. Objective of this study was to identify the factors that influence adoption of agroforestry technologies by farmers in the district. Random sampling was employed to select two kebele administrations and respondents. Data collection was conducted by rural household questionnaire survey, participatory rural appraisal, questionnaires for local and woreda extension staff, secondary data resources and field observation. A sample of 12 key informants, 6 extension staffs, and 182 households, were used in the data collection. Chi square test used to determine significant relationships between adoption of agroforestry and 15 selected variables. Out of which eleven were found to be significant to affect farmers’ adoptiveness. These were frequency of visits of farmers (13.39%), participation in training (11.49%), farmers’ attitude towards agroforestry practices (10.61%), frequency of visits of extensionists (10.38%), participation in extension meeting (10.34%), participation in field day (10.28%), land holding size (9.29%), level of literacy (8.78%), awareness about the importance of agroforestry technology packages (7.06%), time taken from their residence to nearest extension (5.04%) and gender of respondents (3.34%). This study also identified various factors that result in low adoption rates of agroforestry including fear of competition, seedling, rainfall and labour shortage, free grazing, financial problem, expecting trees as soil degrader and long span of trees and lack of need ranking. To improve farmers’ adoption, the factors identified should be well addressed by launching a series and recurrent outreach extension program appropriate and suitable to farmers need.

Keywords: farmers attitude, farmers participation, soil degradation, technology packages

Procedia PDF Downloads 142

Authors: Chang-Hui Shen, Paulina Konarzewska

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Cell homeostasis is regulated by vacuolar activity and it has been shown that lipid composition of the vacuole plays an important role in vacuolar function. The major phosphoinositide species present in the vacuolar membrane include phosphatidylinositol 3,5-biphosphate (PI(3,5)P₂) which is generated from PI(3)P controlled by Fab1p. Deletion of FAB1 gene reduce the synthesis of PI(3,5)P₂ and thus result in enlarged or fragmented vacuoles, with neutral vacuolar pH due to reduced vacuolar H⁺-ATPase activity. These mutants also exhibited poor growth at high extracellular pH and in the presence of CaCl₂. Conversely, VPS34 regulates the synthesis of PI(3)P from phosphatidylinositol (PI), and the lack of Vps34p results in the reduction of vacuolar activity. Although the cellular observations are clear, it is still unknown about the molecular mechanism between the phospholipid biosynthesis pathway and vacuolar activity. Since both VPS34 and FAB1 are important in vacuolar activity, we hypothesize that the molecular mechanism of vacuolar function might be regulated by the transcriptional regulators of phospholipid biosynthesis. In this study, we study the role of the major phospholipid biosynthesis transcription factor, INO2, in the regulation of vacuolar activity. We first performed qRT-PCR to examine the effect of Ino2p on the expression of VPS34 and FAB1. Our results showed that VPS34 was upregulated in the presence of inositol for both WT and ino2Δ cells. However, FAB1 was only upregulated significantly in ino2Δ cells. This indicated that Ino2p might be the negative regulator for FAB1 expression. Next, growth sensitivity experiment showed that WT, vma3Δ, and ino2Δ grew well in growth medium buffered to pH 5.5 containing 10 mM CaCl₂. As cells were switched to growth medium buffered to pH 7 containing CaCl₂ WT, ino2Δ and opi1Δ showed growth reduction, whereas vma3Δ was completely nonviable. As the concentration of CaCl₂ was increased to 60 mM, ino2Δ cells showed moderate growth reduction compared to WT. This result suggests that ino2Δ cells have better vacuolar activity. Microscopic analysis and vacuolar acidification were employed to further elucidate the importance of INO2 in vacuolar homeostasis. Analysis of vacuolar morphology indicated that WT and vma3Δ cells displayed vacuoles that occupied a small area of the cell when grown in media buffered to pH 5.5. Whereas, ino2Δ displayed fragmented vacuoles. On the other hand, all strains grown in media buffered to pH 7, exhibited enlarged vacuoles that occupied most of the cell’s surface. This indicated that the presence of INO2 may play negative effect in vacuolar morphology when cells are grown in media buffered to pH 5.5. Furthermore, vacuolar acidification assay showed that only vma3Δ cells displayed notably less acidic vacuoles as cells were grown in media buffered to pH 5.5 and pH 7. Whereas, ino2Δ cells displayed more acidic pH compared to WT at pH7. Taken together, our results demonstrated the molecular mechanism of the vacuolar activity regulated by the phospholipid biosynthesis transcription factors Ino2p. Ino2p negatively regulates vacuolar activity through the expression of FAB1.

Keywords: vacuole, phospholipid, homeostasis, Ino2p, FAB1

Procedia PDF Downloads 118