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Authors: Ghazi R. Reda Mahmoud Reda

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Among the metal that forms hydride´s, Mg and Ti are known as the most lightweight materials; however, they are covered with a passive layer of oxides and hydroxides and require activation treatment under high temperature ( > 300 C ) and hydrogen pressure ( > 3 MPa) before being used for storage and transport applications. It is well known that small graphite addition to Ti or Mg, lead to a dramatic change in the kinetics of mechanically induced hydrogen sorption ( uptake) and significantly stimulate the Ti-Hydrogen interaction. Many explanations were given by different authors to explain the effect of graphite addition on the performance of Ti as material for hydrogen storage. Not only graphite but also the addition of a polycyclic aromatic compound will also improve the hydrogen absorption kinetics. It will be shown that the function of carbon addition is two-fold. First carbon acts as a vacuum cleaner, which scavenges out all the interstitial oxygen that can poison or slow down hydrogen absorption. It is also important to note that oxygen favors the chemisorption of hydrogen, which is not desirable for hydrogen storage. Second, during scavenging of the interstitial oxygen, the carbon reacts with oxygen in the nano and microchannel through a highly exothermic reaction to produce carbon dioxide and monoxide which provide the necessary heat for activation and thus in the presence of carbon lower heat of activation for hydrogen absorption which is observed experimentally. Furthermore, the product of the reaction of hydrogen with the carbon oxide will produce water which due to ball milling hydrolyze to produce the linear H5O2 + this will reconstruct the primary structure of the nanocarbon to form secondary structure, where the primary structure (a sheet of carbon) are connected through hydrogen bonding. It is the space between these sheets where physisorption or defect mediated sorption occurs.

Keywords: metal forming hydrides, polar molecule impurities, titanium, phase diagram, hydrogen absorption

Procedia PDF Downloads 363

Authors: Ramatu I. Sha’aba, Mathias A. Chia, Abdullahi B. Alhassan, Yisa A. Gana, Ibrahim M. Gadzama

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Globally, Diclofenac (DLC) and Ibuprofen (IBU) are the most prescribed drugs due to their antipyretic and analgesic properties. They are, however, highly toxic at elevated doses, with the involvement of an already described oxidative stress pathway. As a result, there is rising concern about the ecological fate of analgesics on non-target organisms such as Daphnia magna and Phytoplankton species. Phytoplankton is a crucial component of the aquatic ecosystem that serves as the primary producer at the base of the food chain. However, the increasing presence and levels of micropollutants such as these analgesics can disrupt their community structure, dynamics, and ecosystem functions. This study presents a comprehensive series of the physiology, antioxidant response, immobilization, and risk assessment of Diclofenac and Ibuprofen’s effects on Daphnia magna and the Phytoplankton community using a laboratory approach. The effect of DLC and IBU at 27.16 µg/L and 20.89 µg/L, respectively, for a single exposure and 22.39 µg/L for combined exposure of DLC and IBU for the experimental setup. The antioxidant response increased with increasing levels of stress. The highest stressor to the organism was 1000 µg/L of DLC and 10,000 µg/L of IBU. Peroxidase and glutathione -S-transferase activity was higher for Diclofenac + Ibuprofen. The study showed 60% and 70% immobilization of the organism at 1000 g L-1 of DLC and IBU. The two drugs and their combinations adversely impacted Phytoplankton biomass with increased exposure time. However, combining the drugs resulted in more significant adverse effects on physiological and pigment content parameters. The risk assessment calculation for the risk quotient and toxic unit of the analgesic reveals from this study was RQ Diclofenac = 8.41, TU Diclofenac = 3.68, and RQ Ibuprofen = 718.05 and TU Ibuprofen = 487.70. Hence, these findings demonstrate that the current exposure concentrations of Diclofenac and Ibuprofen can immobilize D. magna. This study shows the dangers of multiple drugs in the aquatic environment because their combinations could have additive effects on the structure and functions of Phytoplankton and are capable of immobilizing D. magna.

Keywords: algae, analgesic drug, daphnia magna, toxicity

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Authors: Javier Carroquino, Nieves García-Casarejos, Pilar Gargallo

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Climate change, depletion of non-renewable resources in the current energies, pollution from them, the greater ecological awareness of the population, are factors that suggest the change of energy sources in business. The agri-food industry is a growth sector, concerned about product innovation, process and with a clear awareness of what climate change may mean for it. This sector is supposed to have a high receptivity to the implementation of clean energy, as this favors not only the environment but also the essence of its business. This work, through surveys, aims to know the willingness of Spanish wineries to implement renewable energies in their vineyards, as well as the limitations they perceive for their implementation. This questionnaire allows the characterization of the sector in terms of its geographical typologies, their activity levels, their perception of environmental issues, the degree of implementation of measures to mitigate climate change and improve energy efficiency, and its uses and energy consumption. The analysis of data proves that the penetration of renewable energies is still at low levels, being the most used energies, solar thermal, photovoltaic and biomass. The initial investment seems to be at the origin of the lack of implantation of this type of energy in the wineries, and not so much the costs of operations and maintenance. The environmental management of the wineries is still at an embryonic stage within the company's organization chart, because these services are either outsourced or, if technicians are available, they are not exclusively dedicated to these tasks. However, there is a strong environmental awareness, as evidenced by the number of climate change mitigation and energy efficiency measures already adopted. The gap between high awareness and low achievement is probably due to the lack of knowledge about how to do it or the perception of a high cost.

Keywords: survey, renewable energy, winery, Spanish case

Procedia PDF Downloads 252

Authors: Ceyda Okudu, Secil Eroglu, Khandakar A. S. M. Saadat, Sibel O. Balci

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Ring Finger Protein 2 (RNF2) is a member of polycomb repressive complex 1 (PRC1), which is one of the epigenetic regulators in the genome. When RNF2 combines with other PRC1 members, it mediates the mono-ubiquitination of Histon2A (H2A). In breast cancer, RNF2 is commonly overexpressed, and also it promotes metastasis and invasion in other aggressive tumors like melanoma, prostate, and hepatocarcinoma. The role of RNF2 in the metastasis and invasion of breast cancer has not yet been elucidated. Our aim is to observe the role of RNF2 in metastasis and invasion in this study by miRNA mediated RNF2 gene silencing in breast cancer cell lines. We selected miRNAs, targeting to RNF2 by searching online databases. miR-17-5p, miR20a-5p, and miR-106b-5p were transfected to breast cancer cell lines (MCF-7, MDA-MB-231, SK-BR-3, and ZR-75-1), and also we used normal breast epithelial cell line (hTERT-HME1) to compare RNF2 gene expression level. After 48-72 hours post-transfection, mRNAs were isolated from the cells, and gene expressions were measured by RT-qPCR after from cDNA syntheses. We observed that RNF2 was highly expressed in SK-BR-3 and MDA-MB-231 cell lines opposite to MCF-7 and ZR-75-1 cell lines. RNF2 was downregulated 5, 5 and 7 fold by miR17-5p, miR20a-5p and miR106b-5p respectively in MCF-7. However, in SK-BR-3 and ZR-75-1 cell lines, miRNAs did not affect significantly RNF2 gene expression level. miR20a-5p decreased RNF2 3 fold and miR17-5p and miR106b-5p did not affect MDA-MB-231. After gene expression analysis, we performed metastasis and invasion assay in MCF-7 cells. For metastasis, we used both wound healing assay and Transwell Cell Migration Assay, and we used Transwell Cell Invasion Assay for invasion. The data of this assay showed that miR17-5p and miR20a-5p decreased both invasion and metastasis level, but miR106b-5p has no effect. We would like to conclude that RNF2 can be targeted by miR17-5p, miR20a-5p and miR106b-5p in MCF-7 cells and also RNF2, which is one of the upregulated genes in aggressive tumor, can be decreased by using these miRNAs. In future, we would like to confirm these results at the protein level and also whether these miRNAs are direct target of RNF2 or not.

Keywords: breast cancer, epigenetic, microRNAs, RNF2

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Authors: Fares A. Mayia, Mahmoud A. Yamany, Mushabbab A. Asiri

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In recent years, non-invasive Focused Ultrasound (FU) has been utilized for generating bubbles (cavities) to ablate target tissue by mechanical fractionation. Intensities >10 kW/cm² are required to generate the inertial cavities. The generation, rapid growth, and collapse of these inertial cavities cause tissue fractionation and the process is called Histotripsy. The ability to fractionate tissue from outside the body has many clinical applications including the destruction of the tumor mass. The process of tissue fractionation leaves a void at the treated site, where all the affected tissue is liquefied to particles at sub-micron size. The liquefied tissue will eventually be absorbed by the body. Histotripsy is a promising non-invasive treatment modality. This paper presents a technique for generating inertial cavities at lower intensities (< 1 kW/cm²). The technique (patent pending) is based on amplitude modulation (AM), whereby a low frequency signal modulates the amplitude of a higher frequency FU wave. Cavitation threshold is lower at low frequencies; the intensity required to generate cavitation in water at 10 kHz is two orders of magnitude lower than the intensity at 1 MHz. The Amplitude Modulation technique can operate in both continuous wave (CW) and pulse wave (PW) modes, and the percentage modulation (modulation index) can be varied from 0 % (thermal effect) to 100 % (cavitation effect), thus allowing a range of ablating effects from Hyperthermia to Histotripsy. Furthermore, changing the frequency of the modulating signal allows controlling the size of the generated cavities. Results from in vitro work demonstrate the efficacy of the new technique in fractionating soft tissue and solid calcium carbonate (Chalk) material. The technique, when combined with MR or Ultrasound imaging, will present a precise treatment modality for ablating diseased tissue without affecting the surrounding healthy tissue.

Keywords: focused ultrasound therapy, histotripsy, inertial cavitation, mechanical tissue ablation

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Authors: M. Rizwan, S. Ahmad, U. Farooq, U. Mahmood, S. U. Rehman, P. Akhtar

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This experiment was conducted to investigate the effect of supplementation of herbal and mineral mixture on growth performance of boilers. One hundred and eighty birds were randomly distributed into 6 experimental units of 3 replicates (10 birds/replicate) as: negative control (basal diet), positive control (Lincomycin at the rate of 5g/bag), commercially available herbal-mineral product FitFat™ at the rate of 150g/bag and 200g/bag, and herbal-mineral mixture at the rate of 150g/bag and herbal-mineral mixture at the rate of 300g/bag. The data regarding weekly feed intake, body weight gain and feed conversion ratio were recorded, and fecal samples were collected at the end of starter and finisher phase for nutrient digestibility trial. The results of body weight gain showed significant (P < 0.05) differences in 3rd week of age (506.90g), also, feed intake showed significant (P < 0.05) results in 1st (297.22g), 3rd (936.7g) and 4th (967.8g) week and feed conversion ratio indicated significant (P < 0.05) variations in 1st (1.14) and 3rd (1.74) week of age. The starter phase indicated significant (P < 0.05) differences among all treatments groups in body weight gain (902.2g), feed intake (1843.9g) and feed conversion ratio (1.78). In case of nutrient digestibility trial, results showed significant (P < 0.05) values of dry matter, crude protein, and crude fat in starter phase as 77.74%, 69.37%, and 61.18% respectively and 77.65%, 68.79% and 61.03% respectively, in finisher phase. Based on overall results, it was concluded that the dietary inclusion of combination of herbs and mineral can increase the production performance of broilers.

Keywords: herbal blend, minerals, crop filling, nutrient digestibility, broiler

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Authors: E. Hovad, J. H. Walther, P. Larsen, J. Thorborg, J. H. Hattel

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In sand casting of metal parts for the automotive industry such as brake disks and engine blocks, the molten metal is poured into a sand mold to get its final shape. The DISAMATIC molding process is a way to construct these sand molds for casting of steel parts and in the present work numerical simulations of this process are presented. During the process green sand is blown into a chamber and subsequently squeezed to finally obtain the sand mould. The sand flow is modelled with the Discrete Element method (DEM) and obtaining the correct material parameters for the simulation is the main goal. Different tests will be used to find or calibrate the DEM parameters needed; Poisson ratio, Young modulus, rolling friction coefficient, sliding friction coefficient and coefficient of restitution (COR). The Young modulus and Poisson ratio are found from compression tests of the bulk material and subsequently used in the DEM model according to the Hertz-Mindlin model. The main focus will be on calibrating the rolling resistance and sliding friction in the DEM model with respect to the behavior of “real” sand piles. More specifically, the surface profile of the “real” sand pile will be compared to the sand pile predicted with the DEM for different values of the rolling and sliding friction coefficients. When the DEM parameters are found for the particle-particle (sand-sand) interaction, the particle-wall interaction parameter values are also found. Here the sliding coefficient will be found from experiments and the rolling resistance is investigated by comparing with observations of how the green sand interacts with the chamber wall during experiments and the DEM simulations will be calibrated accordingly. The coefficient of restitution will be tested with different values in the DEM simulations and compared to video footages of the DISAMATIC process. Energy dissipation will be investigated in these simulations for different particle sizes and coefficient of restitution, where scaling laws will be considered to relate the energy dissipation for these parameters. Finally, the found parameter values are used in the overall discrete element model and compared to the video footage of the DISAMATIC process.

Keywords: discrete element method, physical properties of materials, calibration, granular flow

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Authors: Kevin T. Inks

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India’s Brahmaputra River Valley has, since independence, experienced consistent low-intensity guerrilla warfare between ethnic and religious groups. These groups are often organized around perceived ethnic territoriality, and target civilians, communities, and especially migrants belonging to other ethnic and religious groups. Intense flooding and erosion have led to widespread displacement, and disaster relief funds are largely tied to legal land tenure. Displaced residents of informal settlements receive little or no resettlement aid, and their subsequent migration strategies and risk from guerrilla violence are poorly understood. Semi-structured interviews and comprehensive surveys focused on perceptions of risk, efficacy of disaster relief, and migration and adaptation strategies were conducted with households identified as being ‘at-risk’ of catastrophic flooding and erosion in Majuli District, Assam. Interviews with policymakers and government workers were conducted to assess disaster relief efforts in informal settlements, and remote sensing methods were used to identify informal settlement and hydrogeomorphic change. The results show that various ethnic and religious groups have differential strategies and preferences for resettlement. However, these varying strategies are likely to lead to differential levels of risk from guerrilla violence. Members of certain ethnic groups residing in informal settlements, in the absence of resettlement assistance, are more likely to seek out unofficial settlement on land far from the protection of the state and experience greater risk of becoming victims of political violence. As climate change and deforestation are likely to increase the severity of the displacement crisis in the Brahmaputra River Valley, more comprehensive disaster relief and surveying efforts are vital for limiting migration and informal settlement in potential sites of guerrilla warfare.

Keywords: climate, displacement, flooding, India, violence

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Authors: Eman M. Morgan, Hayam M. Lotfy, Yasmin M. Fayez, Mohamed Abdelkawy, Engy Shokry

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Two simple, selective, economic, safe, accurate, precise and environmentally friendly methods were developed and validated for the quantitative determination of valaciclovir (VAL) in the presence of its related substances R1 (acyclovir), R2 (guanine) in bulk powder and in the commercial pharmaceutical product containing the drug. Method A is a colorimetric method where VAL selectively reacts with ferric hydroxamate and the developed color was measured at 490 nm over a concentration range of 0.4-2 mg/mL with percentage recovery 100.05 ± 0.58 and correlation coefficient 0.9999. Method B is a reversed phase ultra performance liquid chromatographic technique (UPLC) which is considered superior in technology to the high-performance liquid chromatography with respect to speed, resolution, solvent consumption, time, and cost of analysis. Efficient separation was achieved on Agilent Zorbax CN column using ammonium acetate (0.1%) and acetonitrile as a mobile phase in a linear gradient program. Elution time for the separation was less than 5 min and ultraviolet detection was carried out at 256 nm over a concentration range of 2-50 μg/mL with mean percentage recovery 100.11±0.55 and correlation coefficient 0.9999. The proposed methods were fully validated as per International Conference on Harmonization specifications and effectively applied for the analysis of valaciclovir in pure form and tablets dosage form. Statistical comparison of the results obtained by the proposed and official or reported methods revealed no significant difference in the performance of these methods regarding the accuracy and precision respectively.

Keywords: hydroxamic acid, related substances, UPLC, valaciclovir

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Authors: Emanuela Chiarella, Clelia Nisticò, Nicola Lombardo, Giovanna Lucia Piazzetta, Nadia Lobello, Maria Mesuraca

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Killian’s Antrochoanal Polyp is a benign lesion of the maxillary sinus characterized by unilateral nasal obstruction, pus discharge, and headache. It affects, more commonly children and young adults. Although its etiology still remains unclear, chronic inflammation, autoreactivity, allergies, and viral infections are strongly associated with its formation and development, resulting in nasal tissue remodeling. We aimed to investigate the stem cells components which reside in this pathological tissue. In particular, we adopted a protocol for the isolation and culturing of mesenchymal stem cells from surgical biopsies of three Killian nasal polyp patients (KNP-MSCs) as well as from their healthy nasal tissue (HNT-MSCs) that were used as controls. The immunophenotype profile of HNT-MSCs and KNP-MSCs was more similar, with a marked positivity for CD73, CD90, and CD105 expression, while being negative for CD34 and CD14 haematopoietic genes. Cell proliferation assay showed that KNP-MSCs had a replicative disadvantage compared to HNT-MSCs, as evidenced by the significantly lower number of cells in the S-phase of the cell cycle. KNP-MSCs also took longer to close a wound than HNT-MSCs, indicating a partial epithelial phenotype in which low levels of ICAM-1 mRNA and a significant increase in E-CAD transcript were detectable. Subsequently, the differentiation potential of both MSCs populations was analyzed by inducing osteoblastic or adipocyte differentiation for up to 20 days. KNP-MSCs showed the ability to differentiate into osteoblasts, although ALP activity as well as the number and size of calcium deposits were lower than osteogenic induced-HNT-MSCs. Also, mRNA levels of osteoblastic marker genes (OCN, OPN, OSX, RUNX2) resulted lower compared to control cell population. Instead, the analysis of the adipogenic differentiation potential showed a similar behavior between KNP-MSCs and HNT-MSCs considering that the amount of lipid droplets, the expression of adipocyte-specific genes (FABP4, AdipoQ, PPARγ2, LPL) and the content of triacylglycerols were almost overlapping. Taken together, these results first demonstrated that Killian's nasal polyp is a source of mesenchymal stem cells with self-renewal and multi-differentiative capabilities.

Keywords: Mesenchymal stem cells, adipogenic differentiation, osteogenic differentiation, EMT

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Authors: Subhasree Majumdar, Sovan Dey, Sayari Mukherjee, Sourav Dutta, Dalia Dasgupta Mandal

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Bacterial isolates from Planococcus genus are known for the production of yellowish orange pigment that belongs to the carotenoid family. These pigments are of immense pharmacological importance as antioxidant, anticancer, eye and liver protective agent, etc. The production of this pigment in a cost effective manner is a challenging task. The present study explored paper mill sludge (PMS), a solid lignocellulosic waste generated in large quantities from pulp and paper mill industry as a substrate for carotenoid pigment production by Planococcus sp. TRC1. PMS was compared in terms of efficacy with sugarcane bagasse, which is a highly explored substrate for valuable product generation via solid state fermentation. The results showed that both the biomasses yielded the highest carotenoid during 48 hours of incubation, 31.6 mg/gm and 42.1 mg/gm for PMS and bagasse respectively. Compositional alterations of both the biomasses showed reduction in lignin, hemicellulose and cellulose content by 41%, 15%, 1% for PMS and 38%, 25% and 6% for sugarcane bagasse after 72 hours of incubation. Structural changes in the biomasses were examined by FT-IR, FESEM, and XRD which further confirmed modification of solid biomasses by bacterial isolate. This study revealed the potential of PMS to act as cheap substrate for carotenoid pigment production by Planococcus sp. TRC1, as it showed a significant production in comparison to sugarcane bagasse which gave only 1.3 fold higher production than PMS. Delignification of PMS by TRC1 during pigment production is another important finding for the reuse of this waste from the paper industry.

Keywords: carotenoid, lignocellulosic, paper mill sludge, Planococcus sp. TRC1, solid state fermentation, sugarcane bagasse

Procedia PDF Downloads 235

Authors: Marthadale Acibo Semacio

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Speaking skill is a crucial part of English language teaching and learning. This actually shows the great importance of this skill in English language classes. Through speaking, ideas and thoughts are shared with other people, and a smooth interaction between people takes place. The study examined the levels of speaking proficiency of the control and experimental groups on pronunciation, grammatical accuracy, and fluency. As a quasi-experimental study, it also determined the presence or absence of significant changes in their speaking proficiency levels in terms of pronouncing the words correctly, the accuracy of grammar and fluency of a language given the two methods to the groups of students in the English language, using the traditional and audio-lingual methods. Descriptive and inferential statistics were employed according to the stated specific problems. The study employed a video presentation with prior information about it. In the video, the teacher acts as model one, giving instructions on what is going to be done, and then the students will perform the activity. The students were paired purposively based on their learning capabilities. Observing proper ethics, their performance was audio recorded to help the researcher assess the learner using the modified speaking rubric. The study revealed that those under the traditional method were more fluent than those in the audio-lingual method. With respect to the way in which each method deals with the feelings of the student, the audio-lingual one fails to provide a principle that would relate to this area and follows the assumption that the intrinsic motivation of the students to learn the target language will spring from their interest in the structure of the language. However, the speaking proficiency levels of the students were remarkably reinforced in reading different words through the aid of aural media with their teachers. The study concluded that using an audio-lingual method of teaching is not a stand-alone method but only an aid of the teacher in helping the students improve their speaking proficiency in the English Language. Hence, audio-lingual approach is encouraged to be used in teaching English language, on top of the chalk-talk or traditional method, to improve the speaking proficiency of students.

Keywords: audio-lingual, speaking, grammar, pronunciation, accuracy, fluency, proficiency

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Authors: Mohamed Abdalla Khalil Mahmoud, Omnia Haidar Suliman

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Information and Communication technologies have revolutionized the way services are developed and offered to customers and have achieved evident success in a variety of vital sectors and widely contributed to the growth and resilience of the economy worldwide. Consequently, governments, especially of developing countries, have turned their attention to examine possible ways to utilize contemporary technology advances to offer essential governmental services to citizens, especially in areas where government agencies are not present. This paper investigates the challenges that impede governments of developing countries to provide basic services to its constituents nationwide. Sudan, as a case study, has taken major steps to provide essential governmental services via electronic channels. However, these services are still not widely used by the citizens, resulting in waste of financial and human resources and efforts that could have been invested more appropriately. This paper examines the challenges that hinder the Sudan’s government in their pursuit of availing its services via electronic channels. Different categories of e-government challenges, such as organizational, technological, social and, demographic, and financial and economic, have been explored in order to pinpoint the major challenges. A structured questionnaire is used to survey the target population of e-government professionals and executives who have direct involvement in the implementation of this nationwide endeavor in Sudan. The survey has successfully identified the main challenges that have high impact on the government’s effort to offer its services via electronic channels, such as Lack of coordination between public and private sectors and Lack of the benefits recognition of the e-government program. The findings of this paper can be used as a solid foundation for improving the way governmental services are offered to citizens in Sudan, resulting in a successful investment of financial and human resources and benefiting the targeted customers of all types.

Keywords: citizen, digital, e-channels, public sector, Sudan, technology

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Authors: Brahim Sow

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Background: Throughout West Africa, Balanites aegyptiaca (BA), or Zygophyllaceae, is widely used in traditional medicine to treat diabetes, hypertension, inflammation, malaria and liver disorders. In our recent research, we found that BA has nephroprotective potential against diabetes mellitus, hypertension and kidney disorders. However, to our knowledge, no systematic studies have been carried out on its derivative (toxicity) profile. Aim of the study: The study was conducted to assess the potential potency of the hydroalcoholic extract of BA bark in rats by the acute and sub-acute oral route. Materials and methods: Male and female rats in the acute depression study received BA extract orally at single doses of 500 mg/kg, 2000 mg/kg, 3000 mg/kg and 5000 mg/kg (n = 6 per group/sex). To assess acute depression, abnormal behaviour, toxic symptoms, weight and death were observed for 14 consecutive days. For the subacute impairment study, Wistar rats received the extract orally at doses of 125, 250 and 500 mg/kg (n=6 per group/sex) per day for 28 days. Behaviour and body weight were monitored daily. At the end of the treatment period, biochemical, haematological and histopathological examinations were performed, and gross and histopathological examinations of several organs were carried out. To determine the presence or absence of phytochemicals, the BA extract was subjected to gage phage chromatographic examination. Results: The absence of absorption chromatography of BA indicates the absence of cyanide groups. This suggests that the BA extract does not contain toxic substances. No mortality or adverse effects were observed at 5000 mg/kg in the acute depression test. With regard to body weight, general behaviour, relative organ weights, haematological and biochemical parameters, BA extract did not induce any mortality or potentially treatment-related effects in the sub-acute study. The normal architecture of the vital organs was revealed by histopathological examination, indicating the absence of morphological alterations. Conclusion: BA extract administered orally for 28 days at doses up to 500 mg/kg did not cause toxicological damage in rats in the present study. The median lethal dose (LD50) of the extract was estimated to be over 5000 mg/kg in an acute hyperglycaemia study.

Keywords: Balanites aegyptiaca L Delile, haematology, biochemistry, rat

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Authors: Hung-Sheng Lin, Cheng-Hsuan Li

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Over the past few years, kernel-based algorithms have been widely used to extend some linear feature extraction methods such as principal component analysis (PCA), linear discriminate analysis (LDA), and nonparametric weighted feature extraction (NWFE) to their nonlinear versions, kernel principal component analysis (KPCA), generalized discriminate analysis (GDA), and kernel nonparametric weighted feature extraction (KNWFE), respectively. These nonlinear feature extraction methods can detect nonlinear directions with the largest nonlinear variance or the largest class separability based on the given kernel function. Moreover, they have been applied to improve the target detection or the image classification of hyperspectral images. The double nearest proportion feature extraction (DNP) can effectively reduce the overlap effect and have good performance in hyperspectral image classification. The DNP structure is an extension of the k-nearest neighbor technique. For each sample, there are two corresponding nearest proportions of samples, the self-class nearest proportion and the other-class nearest proportion. The term “nearest proportion” used here consider both the local information and other more global information. With these settings, the effect of the overlap between the sample distributions can be reduced. Usually, the maximum likelihood estimator and the related unbiased estimator are not ideal estimators in high dimensional inference problems, particularly in small data-size situation. Hence, an improved estimator by shrinkage estimation (regularization) is proposed. Based on the DNP structure, LDA is included as a special case. In this paper, the kernel method is applied to extend DNP to kernel-based DNP (KDNP). In addition to the advantages of DNP, KDNP surpasses DNP in the experimental results. According to the experiments on the real hyperspectral image data sets, the classification performance of KDNP is better than that of PCA, LDA, NWFE, and their kernel versions, KPCA, GDA, and KNWFE.

Keywords: feature extraction, kernel method, double nearest proportion feature extraction, kernel double nearest feature extraction

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Authors: A. Pornpitakdumrong

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The aim of this research study was to develop recipe of Karanda ice cream as healthy promoting ice cream by high protein, low fat and naturally raw material, which found in local area. The results were found that appropriate condition for Karanda ice cream including incubation period, temperature and frozen time, which were 8-12 hours, -20 to -25 °C and 2-4 hours, respectively. Small fruit variety Karanda should selected only ripe fruits for Karanda ice cream made. Because of unripe fruits were contained resin and need to be air dried for reducing level of resin. Therefore, large fruit variety Karanda can be use both ripe and unripe fruits for Karanda ice cream made by without any astringent and bitter taste. However, small fruit variety Karanda was proper to made ice cream for trade, because occurring of industry to select the ripe fruits and commercially frozen, which be providing for the whole year compared with large variety fruits were rarely, low harvesting amount and short shelf life. Karanda ice cream produced from flesh part was attractive but was not accepted by consumers. It may due to resin contained with Karanda pulp, which led to be rough texture of ice cream. We were choose only Karanda juice, which was more appropriated and used Karanda juice with water by 1:1 ratio, because undiluted juice was sour taste. Most acceptance recipe of karanda ice cream product was sixth recipe by 91% of consumers, which was contained soy protein to made ice cream was delicate and swell, milk powder (little amount) to made ice cream was greasy, corn powder as stabilizer and undiluted coconut milk (little amount) to improve ice cream odor and similar to apricot odor.

Keywords: karonda fruits, Carissa carandas Linn, ice cream, healthy ice cream

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Authors: Jeongil Kim

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The present study examined the effects of arts-mediated mother-child dyads mindfulness-based intervention for Korean children with attention deficit hyperactivity disorder (ADHD) and their mothers, on behaviors in children and subjective psychological states in mothers. Four elementary school boys with ADHD and their mothers participated in the study. Using a multiple baseline design across four mother-child dyads, data were collected on the target behaviors (disruptive behavior, on-task behavior, and compliance in class) in children using a 10-second partial interval recording system and on the subjective psychological states in mothers using four questionnaires (on perceived stress, burnout, mindfulness, and satisfaction with life). The intervention consisted of a) mindfulness training, b) mindfulness practice, and c) mindful management of body and feeling. The arts activities, making a coiled clay pot and Korean traditional music performance, were utilized to facilitate the environment to help each participant to understand the content and progress of the intervention program. The results showed that all four dyads showed improvement in adaptive behaviors in the children (increase in on-task behavior; decrease in disruptive behavior) and positive change in subjective psychological states in the mothers (increase in scores of mindfulness and satisfaction with life; decrease in scores of perceived stress and burnout). The changes in the children’s behaviors and in the mothers’ subjective psychological states were maintained when the intervention was drawn and generalized in novel settings. The results suggest that arts-mediated mother-child dyads mindfulness-based intervention would be a mutual benefiting strategy to support both children with ADHD and their mothers who experience diverse challenges in behavioral and psychological aspects.

Keywords: mindfulness, attention deficit hyperactivity disorder (ADHD), arts-mediated, behavior, psychological well-being, child-mother

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Authors: Nounamo Nguedie Yann Harold

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The factors behind the debt have been the subject of several studies in the literature. Pioneering studies based on the 'double deficit' approach linked indebtedness to the imbalance between savings and investment, the budget deficit and the current account deficit. Most studies on identifying factors that may stimulate or reduce the level of external public debt agree that the following variables are important explanatory variables in leveraging debt: the budget deficit, trade opening, current account and exchange rate, import, export, interest rate, term variation exchange rate, economic growth rate and debt service, capital flight, and over-indebtedness. Few studies addressed the impact of political factors on the level of external debt. In general, however, the IMF's stabilization programs in developing countries following the debt crisis have resulted in economic recession and the advent of political crises that have resulted in changes in governments. In this sense, political institutions are recognised as factors of accumulation of external debt in most developing countries. This paper assesses the role of political factors on the external debt level of African countries in the Franc Zone over the period 1985-2016. Data used come from World Bank and ICRG. Using a logit in panel, the results show that the more a country is politically stable, the lower the external debt compared to the gross domestic product. Political stability multiplies 1.18% the chances of being in the sustainable debt zone. For example, countries with good political institutions experience less severe external debt burdens than countries with bad political institutions.

Keywords: African countries, external debt, Franc Zone, political factors

Procedia PDF Downloads 219

Authors: Nakul V. Naphade, Sandra G. L. Persiani, Yew Wah Wong, Pramod S. Kamath, Avinash H. Anantharam, Hui Ling Aw, Yann Grynberg

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Achieving zero-energy targets in existing buildings is known to be a difficult task requiring important cuts in the building energy consumption, which in many cases clash with the functional necessities of the building wherever the on-site energy generation is unable to match the overall energy consumption. Between the building’s consumption optimization limit and the energy, target stretches a case-specific optimization grey zone, which requires tailored intervention and enhanced user’s commitment. In the view of the future adoption of more stringent energy-efficiency targets in the context of hot-humid tropical climates, this study aims to define the energy optimization grey zone by assessing the energy-efficiency limit in the state-of-the-art typical mid- and high-rise full AC office buildings, through the integration of currently available technologies. Energy models of two code-compliant generic office-building typologies were developed as a baseline, a 20-storey ‘high-rise’ and a 7-storey ‘mid-rise’. Design iterations carried out on the energy models with advanced market ready technologies in lighting, envelope, plug load management and ACMV systems and controls, lead to a representative energy model of the current maximum technical potential. The simulations showed that ZEB targets could be achieved in fully AC buildings under an average of seven floors only by compromising on energy-intense facilities (as full AC, unlimited power-supply, standard user behaviour, etc.). This paper argues that drastic changes must be made in tropical buildings to span the energy optimization grey zone and achieve zero energy. Fully air-conditioned areas must be rethought, while smart technologies must be integrated with an aggressive involvement and motivation of the users to synchronize with the new system’s energy savings goal.

Keywords: energy simulation, office building, tropical climate, zero energy buildings

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Authors: Shreya Srivastava, Sagnik Dey

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Aerosol direct radiative forcing (ADRF) refers to the alteration of the Earth's energy balance from the scattering and absorption of solar radiation by aerosol particles. India experiences substantial ADRF due to high aerosol loading from various sources. These aerosols' radiative impact depends on their physical characteristics (such as size, shape, and composition) and atmospheric distribution. Quantifying ADRF is crucial for understanding aerosols’ impact on the regional climate and the Earth's radiative budget. In this study, we have taken radiation data from Clouds and the Earth’s Radiant Energy System (CERES, spatial resolution=1ox1o) for 22 years (2000-2021) over the Indian subcontinent. Except for a few locations, the short-wave DARF exhibits aerosol cooling at the TOA (values ranging from +2.5 W/m2 to -22.5W/m2). Cooling due to aerosols is more pronounced in the absence of clouds. Being an aerosol hotspot, higher negative ADRF is observed over the Indo-Gangetic Plain (IGP). Aerosol Forcing Efficiency (AFE) shows a decreasing seasonal trend in winter (DJF) over the entire study region while an increasing trend over IGP and western south India during the post-monsoon season (SON) in clear-sky conditions. Analysing atmospheric heating and AOD trends, we found that only the aerosol loading is not governing the change in atmospheric heating but also the aerosol composition and/or their vertical profile. We used a Multi-angle Imaging Spectro-Radiometer (MISR) Level-2 Version 23 aerosol products to look into aerosol composition. MISR incorporates 74 aerosol mixtures in its retrieval algorithm based on size, shape, and absorbing properties. This aerosol mixture information was used for analysing long-term changes in aerosol composition and dominating aerosol species corresponding to the aerosol forcing value. Further, ADRF derived from this method is compared with around 35 studies across India, where a plane parallel Radiative transfer model was used, and the model inputs were taken from the OPAC (Optical Properties of Aerosols and Clouds) utilizing only limited aerosol parameter measurements. The result shows a large overestimation of TOA warming by the latter (i.e., Model-based method).

Keywords: aerosol radiative forcing (ARF), aerosol composition, MISR, CERES, SBDART

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Authors: Carlos Roberto De Rolt, Julio da Silva Dias, Rafael Tezza, Luca Foschini, Matteo Mura

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The seasonal population fluctuation presents a challenge to touristic cities since the number of inhabitants can double according to the season. The aim of this work is to develop a model that correlates the waste collected with the population of the city and also allow cooperation between the inhabitants and the local government. The model allows public managers to evaluate the impact of the seasonal population fluctuation on waste generation and also to improve planning resource utilization throughout the year. The study uses data from the company that collects the garbage in Florianópolis, a Brazilian city that presents the profile of a city that attracts tourists due to numerous beaches and warm weather. The fluctuations are caused by the number of people that come to the city throughout the year for holidays, summer time vacations or business events. Crowdsensing will be accomplished through smartphones with access to an app for data collection, with voluntary participation of the population. Crowdsensing participants can access information collected in waves for this portal. Crowdsensing represents an innovative and participatory approach which involves the population in gathering information to improve the quality of life. The management of crowdsensing solutions plays an essential role given the complexity to foster collaboration, establish available sensors and collect and process the collected data. Practical implications of this tool described in this paper refer, for example, to the management of seasonal tourism in a large municipality, whose public services are impacted by the floating of the population. Crowdsensing and big data support managers in predicting the arrival, permanence, and movement of people in a given urban area. Also, by linking crowdsourced data to databases from other public service providers - e.g., water, garbage collection, electricity, public transport, telecommunications - it is possible to estimate the floating of the population of an urban area affected by seasonal tourism. This approach supports the municipality in increasing the effectiveness of resource allocation while, at the same time, increasing the quality of the service as perceived by citizens and tourists.

Keywords: big data, dashboards, floating population, smart city, urban management solutions

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Authors: Ergün Şakalar, Şeyma Özçirak Ergün, Emrah Yalazi̇, Emine Altinkaya, Cengiz Ataşoğlu

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In recent years, food allergies which cause serious health problems affect to public health around the world. Foodstuffs which contain allergens are either intentionally used as ingredients or are encased as contaminant in food products. The prevalence of clinical allergy to peanuts and nuts is estimated at about 0.4%-1.1% of the adult population, representing the allergy to pistachio the 7% of the cases of tree nut causing allergic reactions. In order to protect public health and enforce the legislation, methods for sensitive analysis of pistachio and peanut contents in food are required. Pea, pistachio and peanut are used together, to reduce the cost in food production such as baklava, snack foods.DNA technology-based methods in food analysis are well-established and well-roundedtools for species differentiation, allergen detection. Especially, the probe-based TaqMan real-time PCR assay can amplify target DNA with efficiency, specificity, and sensitivity.In this study, pistachio, peanut and pea were finely ground and three separate series of triplet mixtures containing 0.1, 1, 10, 100, 1000, 10,000 and 100,000 mg kg-1 of each sample were prepared for each series, to a final weight of 100 g. DNA from reference samples and industrial products was successfully extracted with the GIDAGEN® Multi-Fast DNA Isolation Kit. TaqMan probes were designed for triplex determination of ITS, Ara h 3 and pea lectin genes which are specific regions for identification pistachio, peanut and pea, respectively.The real-time PCR as quantitative detected pistachio, peanut and pea in these mixtures down to the lowest investigated level of 0.1, 0.1 and 1 mg kg-1, respectively. Also, the methods reported here are capable of detecting of as little as 0.001% level of peanut DNA, 0,000001% level of pistachio DNA and 0.000001% level of pea DNA. We accomplish that the quantitative triplex real-time PCR method developed in this study canbe applied to detect pistachio, peanut and peatraces for three allergens at once in commercial food products.

Keywords: allergens, DNA, real-time PCR, TaqMan probe

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Authors: Suvarna Kumar Gogula, Sandhya Devi Gogula, P. Chanakya

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Augmented reality is defined as the collection of the digital (or) computer generated information like images, audio, video, 3d models, etc. and overlay them over the real time environment. Augmented reality can be thought as a blend between completely synthetic and completely real. Augmented reality provides scope in a wide range of industries like manufacturing, retail, gaming, advertisement, tourism, etc. and brings out new dimensions in the modern digital world. As it overlays the content, it makes the users enhance the knowledge by providing the content blended with real world. In this application, we integrated augmented reality with data analytics and integrated with cloud so the virtual content will be generated on the basis of the data present in the database and we used marker based augmented reality where every marker will be stored in the database with corresponding unique ID. This application can be used in wide range of industries for different business processes, but in this paper, we mainly focus on the marketing industry which helps the customer in gaining the knowledge about the products in the market which mainly focus on their prices, customer feedback, quality, and other benefits. This application also focuses on providing better market strategy information for marketing managers who obtain the data about the stocks, sales, customer response about the product, etc. In this paper, we also included the reports from the feedback got from different people after the demonstration, and finally, we presented the future scope of Augmented Reality in different business processes by integrating with new technologies like cloud, big data, artificial intelligence, etc.

Keywords: augmented reality, data analytics, catch room, marketing and sales

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Authors: Rahat Zaman

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The present study was conducted to investigate psychological well-being and positive and negative affect among adults. The sample comprised 221 adults; the sample was collected from all over Pakistan. Psychological well-being was measured with the help of the psychological well-being scale developed by Ryff and Keyes (1995). Lifestyle was measured with the help of the Health Promoting Lifestyle Profile Scale developed by Walker et al. (1995). Positive and negative effects were measured by PANAS, developed by Watson, Clark, and Tellegen (1998). To check the properties of scale, the alpha reliability coefficient was calculated. To test the hypotheses of the research, correlation, independent sample t-rest, and ANOVA were computed. It was hypothesized that there would be a positive relationship between psychological well-being and lifestyles and positive affect. The results show that psychological well-being, lifestyle, and positive affect are positively related. This also supports our hypothesis. The research also searched for relationships in the study variables according to the demographics of the sample. The respondents varied according to their dominant affect levels with respect to their psychological well-being and lifestyles. The research found significant differences for the genders in life appreciation, nutrition, and negative affect. Single and married individuals differed significantly on autonomy, environmental mastery, life appreciation, nutrition, and stress management. Individuals showed significant differences with respect to their living situation, joint and nuclear family members showed significant differences in personal growth, autonomy, health responsibilities, social support, physical activities, and stress management. The sample showed significant differences in environmental mastery, personal growth, purpose in life, life appreciation, health responsibilities, physical activities, stress management, and negative affect when divided in socioeconomic status. Age-wise analysis showed significant differences in autonomy, personal growth, purpose in life, life appreciation, nutrition, and stress management. Provincially significant differences were found in life appreciation, nutrition, social support, physical activities, and stress management, and both positive and negative effects were experienced. Implications of the results are discussed.

Keywords: wellbeing, healthy lifestyle, self acceptance, positive

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Authors: Soultana Konstantinidou, Tiziana Schmidt, Elena Landi, Alessandro De Carli, Giovanni Maltinti, Darius Witt, Alicja Dziadosz, Agnieszka Lindstaedt, Michele Lai, Mauro Pistello, Valentina Cappello, Luciana Dente, Chiara Gabellini, Piotr Barski, Vittoria Raffa

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CRISPR/Cas9 technology has gained the interest of researchers in the field of biotechnology for genome editing. Since its discovery as a microbial adaptive immune defense, this system has been widely adopted and is acknowledged for having a variety of applications. However, critical barriers related to safety and delivery are persisting. Here, we propose a new concept of genome engineering, which is based on a nano-formulation of Cas9. The Cas9 enzyme was conjugated to a gold nanoparticle (AuNP-Cas9). The AuNP-Cas9 maintained its cleavage efficiency in vitro, to the same extent as the ribonucleoprotein, including non-conjugated Cas9 enzyme, and showed high gene editing efficiency in vivo in zebrafish embryos. Since CRISPR/Cas9 technology is extensively used in cancer research, melanoma was selected as a validation target. Cell studies were performed in A375 human melanoma cells. Particles per se had no impact on cell metabolism and proliferation. Intriguingly, the AuNP-Cas9 internalized spontaneously in cells and localized as a single particle in the cytoplasm and organelles. More importantly, the AuNP-Cas9 showed a high nuclear localization signal. The AuNP-Cas9, overcoming the delivery difficulties of Cas9, could be used in cellular biology and localization studies. Taking advantage of the plasmonic properties of gold nanoparticles, this technology could potentially be a bio-tool for combining gene editing and photothermal therapy in cancer cells. Further work will be focused on intracellular interactions of the nano-formulation and characterization of the optical properties.

Keywords: CRISPR/Cas9, gene editing, gold nanoparticles, nanotechnology

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Authors: Brody R. Clark, Chaminda Gallage, John Yeaman

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Multigrade bitumen asphalt is a quality asphalt product that is not utilised in many places globally. Multigrade bitumen is believed to be less sensitive to temperature, which gives it an advantage over conventional binders. Previous testing has shown that asphalt temperature changes greatly with depth, but currently the industry standard is to nominate a single temperature for design. For detailed design of asphalt roads, perhaps asphalt layers should be divided into nominal layer depths and different modulus and fatigue equations/values should be used to reflect the temperatures of each respective layer. A collaboration of previous laboratory testing conducted on multigrade bitumen asphalt beams under a range of temperatures and loading conditions was analysed. The samples tested included 0% or 15% recycled asphalt pavement (RAP) to determine what impact the recycled material has on the fatigue life and stiffness of the pavement. This paper investigated the temperature susceptibility of multigrade bitumen asphalt pavements compared to conventional binders by combining previous testing that included conducting a sweep of fatigue tests, developing complex modulus master curves for each mix and a study on how pavement temperature changes through pavement depth. This investigation found that the final design of the pavement is greatly affected by the nominated pavement temperature and respective material properties. This paper has outlined a potential revision to the current design approach for asphalt pavements and proposes that further investigation is needed into pavement temperature and its incorporation into design.

Keywords: asphalt, complex modulus, fatigue life, flexural stiffness, four point bending, multigrade bitumen, recycled asphalt pavement

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Authors: G. Ramya

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Value engineering is a systematic approach, widely used to optimize the design or process or product in the designing stage. It used to achieve the client's obligation by increasing the functionality and attain the targeted cost in the cost planning. Value engineering effectiveness and benefits decrease along with the progress of the project since the change in the scope of the work and design will account for more cost all along the lifecycle of the project. Integrating the value engineering with other project management activities will promote cost minimization, client satisfaction, and ensure early completion of the project in time. Previous research studies suggested that value engineering can integrate with other project delivery activities, but research studies unable to frame a model that collaborates the project management activities with the job plan of value engineering approach. I analyzed various project management activities and their synergy between each other. The project management activities and processes like a)risk analysis b)lifecycle cost analysis c)lean construction d)facility management e)Building information modelling f)Contract administration, collaborated, and project delivery model planned along with the RIBA plan of work. The key outcome of the research is a value-driven project delivery model, which will succeed in dealing with the economic impact, constraints and conflicts arise due to the COVID-19 outbreak in the Indian construction sector. Benefits associated with the structured framework is construction project delivery that ensures early contractor involvement, mutual risk sharing, and reviving the project with a cost overrun and delay back on track ,are discussed. Keywords: Value-driven project delivery model, Integration, RIBA plan of work Themes: Design Economics

Keywords: value-driven project delivery model, Integration, RIBA

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Authors: Upasana G. Singh, Meera Gungea

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Learning is no more confined to the traditional classroom, teacher, and student interaction. Many universities offer courses through the Open and Distance Learning (ODL) mode, attracting a diversity of learners in terms of age, gender, and profession to name a few. The ODL mode has surfaced as one of the famous sought-after modes of learning, allowing learners to invest in their educational growth without hampering their personal and professional commitments. This mode of learning, however, requires that those who ultimately choose to adopt it must be prepared to undertake studies through such medium. The purpose of this research is to assess whether students who join universities offering courses through the ODL mode are ready to embark and study within such a framework. This study will be helpful to unveil the challenges students face in such an environment and thus contribute to developing a framework to ease adoption and integration into the ODL environment. Prior to the implementation of e-learning, a readiness assessment is essential for any institution that wants to adopt any form of e-learning. Various e-learning readiness assessment models have been developed over the years. However, this study is based on a conceptual model for e-Learning Readiness Assessment which is a ‘hybrid model’. This hybrid model consists of 4 main parameters: 1) Technological readiness, 2) Culture readiness, 3) Content readiness, and 4) Demographics factors, with 4 sub-areas, namely, technology, innovation, people and self-development. The model also includes the attitudes of users towards the adoption of e-learning as an important aspect of assessing e-learning readiness. For this study, some factors and sub-factors of the hybrid model have been considered and adapted, together with the ‘Attitude’ component. A questionnaire was designed based on the models and students where the target population were students enrolled at the Open University of Mauritius, in undergraduate and postgraduate courses. Preliminary findings indicate that most (68%) learners have an average knowledge about ODL form of learning, despite not many (72%) having previous experience with ODL. Despite learning through ODL 74% of learners preferred hard copy learning material and 48% found difficulty in reading learning material on electronic devices.

Keywords: open learning, distance learning, student readiness, a hybrid model

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Authors: Byron Joseph A. Hallar, Annjeannette Alain D. Galang, Maria Visitacion N. Gumabay

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The widespread adoption and increasing proliferation of smartphones that started during the first decade of the twenty-first century have enabled their users to communicate and access information in ways that were merely thought of as possibilities in the few years before the smartphone revolution. A product of the convergence of the cellular phone and portable computer, the smartphone provides an additional important function that used to be the exclusive domain of desktop-bound computers and portable computers: Web Browsing. For increasing numbers of users, the smartphone and allied devices such as tablet computers have become their first and often their only means of accessing the World Wide Web. This has led to the development of websites that cater to the needs of the new breed of smartphone-carrying web users. The smaller size of smartphones as compared with conventional computers has provided unique challenges to web interface designers. The smaller screen size and touchscreen interface have made it much more difficult to read and navigate through web pages that were in most part designed for traditional desktop and portable computers. Although increasing numbers of websites now provide an alternate website formatted for smartphones, problems with ease of use, reliability and usability still remain. This study focuses on the identification of the problems associated with smartphone web interfaces, the compliance with accepted standards of user-oriented web interface design, the strategies that could be utilized to ensure the design of user-centric web interfaces for smartphones, and the identification of the current trends and developments related to user-centric web interface design intended for the consumption of smartphone users.

Keywords: human-computer interaction, user-centered design, web interface, mobile, smartphone

Procedia PDF Downloads 356

Authors: Asavasereerat K., Teacharsripaitoon T., Tungyingyong P., Charupongrat S., Noppiboon S. Hochareon L., Kitsuban P.

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Third generation of vaccines is based on gene therapy where DNA is introduced into patients. The antigenic or therapeutic proteins encoded from transgenes DNA triggers an immune-response to counteract various diseases. Moreover, DNA vaccine offers the customization of its ability on protection and treatment with high stability. The production of DNA vaccines become of interest. According to USFDA guidance for industry, the recommended limits for impurities from host cell are lower than 1%, and the active conformation homogeneity supercoiled DNA, is more than 80%. Thus, the purification strategy using two-steps chromatography has been established and verified for its robustness. Herein, pVax1/lacZ, a pre-approved USFDA DNA vaccine backbone, was used and transformed into E. coli strain DH5α. Three purification process parameters including sample-loading flow rate, the salt concentration in washing and eluting buffer, were studied and the experiment was designed using response surface method with central composite face-centered (CCF) as a model. The designed range of selected parameters was 10% variation from the optimized set point as a safety factor. The purity in the percentage of supercoiled conformation obtained from each chromatography step, AIEX and HIC, were analyzed by HPLC. The response data were used to establish regression model and statistically analyzed followed by Monte Carlo simulation using SAS JMP. The results on the purity of the product obtained from AIEX and HIC are between 89.4 to 92.5% and 88.3 to 100.0%, respectively. Monte Carlo simulation showed that the pVAX1/lacZ purification process is robust with confidence intervals of 0.90 in range of 90.18-91.00% and 95.88-100.00%, for AIEX and HIC respectively.

Keywords: AIEX, DNA vaccine, HIC, puification, response surface method, robustness

Procedia PDF Downloads 208