Search results for: voice activity detection
5057 Structure-Reactivity Relationship of Some Rhᴵᴵᴵ and Osᴵᴵᴵ Complexes with N-Inert Ligands in Ionic Liquids
Authors: Jovana Bogojeski, Dusan Cocic, Nenad Jankovic, Angelina Petrovic
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Kinetically-inert transition metal complexes, such as Rh(III) and Os(III) complexes, attract increasing attention as leading scaffolds for the development of potential pharmacological agents due to their inertness and stability. Therefore, we have designed and fully characterized a few novel rhodium(III) and osmium(III) complexes with a tridentate nitrogen−donor chelate system. For some complexes, the crystal X-ray structure analysis was performed. Reactivity of the newly synthesized complexes towards small biomolecules, such as L-methionine (L-Met), guanosine-5’-monophosphate (5’-GMP), and glutathione (GSH) has been examined. Also, the reactivity of these complexes towards the DNA/RNA (Ribonucleic acid) duplexes was investigated. Obtained results show that the newly synthesized complexes exhibit good affinity towards the studied ligands. Results also show that the complexes react faster with the RNA duplex than with the DNA and that in the DNA duplex reaction is faster with 15mer GG than with the 22mer GG. The UV-Vis (Ultraviolet-visible spectroscopy) is absorption spectroscopy, and the EB (Ethidium bromide) displacement studies were used to examine the interaction of these complexes with CT-DNA and BSA (Bovine serum albumin). All studied complex showed good interaction ability with both the DNA and BSA. Furthermore, the DFT (Density-functional theory) calculation and docking studies were performed. The impact of the metal complex on the cytotoxicity was tested by MTT assay (a colorimetric assay for assessing cell metabolic activity) on HCT-116 lines (human colon cancer cell line). In addition, all these tests were repeated in the presence of several water-soluble biologically active ionic liquids. Attained results indicate that the ionic liquids increase the activity of the investigated complexes. All obtained results in this study imply that the introduction of different spectator ligand can be used to improve the reactivity of rhodium(III) and osmium(III) complexes. Finally, these results indicate that the examined complexes show reactivity characteristics needed for potential anti-tumor agents, with possible targets being both the DNA and proteins. Every new contribution in this field is highly warranted due to the current lack of clinically used Metallo-based alternatives to cisplatin.Keywords: biomolecules, ionic liquids, osmium(III), rhodium(III)
Procedia PDF Downloads 1515056 Full Characterization of Heterogeneous Antibody Samples under Denaturing and Native Conditions on a Hybrid Quadrupole-Orbitrap Mass Spectrometer
Authors: Rowan Moore, Kai Scheffler, Eugen Damoc, Jennifer Sutton, Aaron Bailey, Stephane Houel, Simon Cubbon, Jonathan Josephs
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Purpose: MS analysis of monoclonal antibodies (mAbs) at the protein and peptide levels is critical during development and production of biopharmaceuticals. The compositions of current generation therapeutic proteins are often complex due to various modifications which may affect efficacy. Intact proteins analyzed by MS are detected in higher charge states that also provide more complexity in mass spectra. Protein analysis in native or native-like conditions with zero or minimal organic solvent and neutral or weakly acidic pH decreases charge state value resulting in mAb detection at higher m/z ranges with more spatial resolution. Methods: Three commercially available mAbs were used for all experiments. Intact proteins were desalted online using size exclusion chromatography (SEC) or reversed phase chromatography coupled on-line with a mass spectrometer. For streamlined use of the LC- MS platform we used a single SEC column and alternately selected specific mobile phases to perform separations in either denaturing or native-like conditions: buffer A (20 % ACN, 0.1 % FA) with Buffer B (100 mM ammonium acetate). For peptide analysis mAbs were proteolytically digested with and without prior reduction and alkylation. The mass spectrometer used for all experiments was a commercially available Thermo Scientific™ hybrid Quadrupole-Orbitrap™ mass spectrometer, equipped with the new BioPharma option which includes a new High Mass Range (HMR) mode that allows for improved high mass transmission and mass detection up to 8000 m/z. Results: We have analyzed the profiles of three mAbs under reducing and native conditions by direct infusion with offline desalting and with on-line desalting via size exclusion and reversed phase type columns. The presence of high salt under denaturing conditions was found to influence the observed charge state envelope and impact mass accuracy after spectral deconvolution. The significantly lower charge states observed under native conditions improves the spatial resolution of protein signals and has significant benefits for the analysis of antibody mixtures, e.g. lysine variants, degradants or sequence variants. This type of analysis requires the detection of masses beyond the standard mass range ranging up to 6000 m/z requiring the extended capabilities available in the new HMR mode. We have compared each antibody sample that was analyzed individually with mixtures in various relative concentrations. For this type of analysis, we observed that apparent native structures persist and ESI is benefited by the addition of low amounts of acetonitrile and formic acid in combination with the ammonium acetate-buffered mobile phase. For analyses on the peptide level we analyzed reduced/alkylated, and non-reduced proteolytic digests of the individual antibodies separated via reversed phase chromatography aiming to retrieve as much information as possible regarding sequence coverage, disulfide bridges, post-translational modifications such as various glycans, sequence variants, and their relative quantification. All data acquired were submitted to a single software package for analysis aiming to obtain a complete picture of the molecules analyzed. Here we demonstrate the capabilities of the mass spectrometer to fully characterize homogeneous and heterogeneous therapeutic proteins on one single platform. Conclusion: Full characterization of heterogeneous intact protein mixtures by improved mass separation on a quadrupole-Orbitrap™ mass spectrometer with extended capabilities has been demonstrated.Keywords: disulfide bond analysis, intact analysis, native analysis, mass spectrometry, monoclonal antibodies, peptide mapping, post-translational modifications, sequence variants, size exclusion chromatography, therapeutic protein analysis, UHPLC
Procedia PDF Downloads 3625055 Investigation of Mutagenicity and DNA Binding Properties of Metal-Free and Metallophthalocyanines Containing α-Napththolbenzein Groups on the Peripheral Positions
Authors: Meltem Betül Sağlam, Halil İbrahim Güler, Aykut Sağlam
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In this work, phthalocyanine compounds containing α-naphtholbenzeinunits have been synthesized. Mutagenicity and DNA binding properties of the compounds were investigated by Salmonella/Microsome Assay and spectrophotometer. According to the results of the preliminary range finding tests, the compounds gave no toxic effect to all tester strain S. typhimurium TA98 and TA100 at doses of 500, 1100, 350, 500 and 750 µg/plate in the presence and absence of S9, respectively. This study showed that all compounds exhibited efficient DNA-binding activity. In conclusion, these non-toxic compounds may be used as effective DNA dyes for molecular biology studies.Keywords: dye, mutagenicity, phthalocyanine, toxicity
Procedia PDF Downloads 2325054 An Optimal Steganalysis Based Approach for Embedding Information in Image Cover Media with Security
Authors: Ahlem Fatnassi, Hamza Gharsellaoui, Sadok Bouamama
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This paper deals with the study of interest in the fields of Steganography and Steganalysis. Steganography involves hiding information in a cover media to obtain the stego media in such a way that the cover media is perceived not to have any embedded message for its unintended recipients. Steganalysis is the mechanism of detecting the presence of hidden information in the stego media and it can lead to the prevention of disastrous security incidents. In this paper, we provide a critical review of the steganalysis algorithms available to analyze the characteristics of an image stego media against the corresponding cover media and understand the process of embedding the information and its detection. We anticipate that this paper can also give a clear picture of the current trends in steganography so that we can develop and improvise appropriate steganalysis algorithms.Keywords: optimization, heuristics and metaheuristics algorithms, embedded systems, low-power consumption, steganalysis heuristic approach
Procedia PDF Downloads 2925053 A Green Method for Selective Spectrophotometric Determination of Hafnium(IV) with Aqueous Extract of Ficus carica Tree Leaves
Authors: A. Boveiri Monji, H. Yousefnia, M. Haji Hosseini, S. Zolghadri
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A clean spectrophotometric method for the determination of hafnium by using a green reagent, acidic extract of Ficus carica tree leaves is developed. In 6-M hydrochloric acid, hafnium reacts with this reagent to form a yellow product. The formed product shows maximum absorbance at 421 nm with a molar absorptivity value of 0.28 × 104 l mol⁻¹ cm⁻¹, and the method was linear in the 2-11 µg ml⁻¹ concentration range. The detection limit value was found to be 0.312 µg ml⁻¹. Except zirconium and iron, the selectivity was good, and most of the ions did not show any significant spectral interference at concentrations up to several hundred times. The proposed method was green, simple, low cost, and selective.Keywords: spectrophotometric determination, Ficus caricatree leaves, synthetic reagents, hafnium
Procedia PDF Downloads 2095052 Khaya Cellulose Supported Copper Nanoparticles for Chemo Selective Aza-Michael Reactions
Authors: M. Shaheen Sarkar, M. Lutfor Rahman, Mashitah Mohd Yusoff
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We prepared a highly active Khaya cellulose supported poly(hydroxamic acid) copper nanoparticles by the surface modification of Khaya cellulose through graft co-polymerization and subsequently amidoximation. The Cu-nanoparticle (0.05 mol% to 50 mol ppm) was selectively promoted Aza-Michael reaction of aliphatic amines to give the corresponding alkylated products at room temperature in methanol. The supported nanoparticle was easy to recover and reused seven times without significance loss of its activity.Keywords: Aza-Michael, copper, cellulose, nanoparticles, poly(hydroxamic acid)
Procedia PDF Downloads 3435051 Photoemission Momentum Microscopy of Graphene on Ir (111)
Authors: Anna V. Zaporozhchenko, Dmytro Kutnyakhov, Katherina Medjanik, Christian Tusche, Hans-Joachim Elmers, Olena Fedchenko, Sergey Chernov, Martin Ellguth, Sergej A. Nepijko, Gerd Schoenhense
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Graphene reveals a unique electronic structure that predetermines many intriguing properties such as massless charge carriers, optical transparency and high velocity of fermions at the Fermi level, opening a wide horizon of future applications. Hence, a detailed investigation of the electronic structure of graphene is crucial. The method of choice is angular resolved photoelectron spectroscopy ARPES. Here we present experiments using time-of-flight (ToF) momentum microscopy, being an alternative way of ARPES using full-field imaging of the whole Brillouin zone (BZ) and simultaneous acquisition of up to several 100 energy slices. Unlike conventional ARPES, k-microscopy is not limited in simultaneous k-space access. We have recorded the whole first BZ of graphene on Ir(111) including all six Dirac cones. As excitation source we used synchrotron radiation from BESSY II (Berlin) at the U125-2 NIM, providing linearly polarized (both polarizations p- and s-) VUV radiation. The instrument uses a delay-line detector for single-particle detection up the 5 Mcps range and parallel energy detection via ToF recording. In this way, we gather a 3D data stack I(E,kx,ky) of the full valence electronic structure in approx. 20 mins. Band dispersion stacks were measured in the energy range of 14 eV up to 23 eV with steps of 1 eV. The linearly-dispersing graphene bands for all six K and K’ points were simultaneously recorded. We find clear features of hybridization with the substrate, in particular in the linear dichroism in the angular distribution (LDAD). Recording of the whole Brillouin zone of graphene/Ir(111) revealed new features. First, the intensity differences (i.e. the LDAD) are very sensitive to the interaction of graphene bands with substrate bands. Second, the dark corridors are investigated in detail for both, p- and s- polarized radiation. They appear as local distortions of photoelectron current distribution and are induced by quantum mechanical interference of graphene sublattices. The dark corridors are located in different areas of the 6 Dirac cones and show chirality behaviour with a mirror plane along vertical axis. Moreover, two out of six show an oval shape while the rest are more circular. It clearly indicates orientation dependence with respect to E vector of incident light. Third, a pattern of faint but very sharp lines is visible at energies around 22eV that strongly remind on Kikuchi lines in diffraction. In conclusion, the simultaneous study of all six Dirac cones is crucial for a complete understanding of dichroism phenomena and the dark corridor.Keywords: band structure, graphene, momentum microscopy, LDAD
Procedia PDF Downloads 3405050 Development of a New Polymeric Material with Controlled Surface Micro-Morphology Aimed for Biosensors Applications
Authors: Elham Farahmand, Fatimah Ibrahim, Samira Hosseini, Ivan Djordjevic, Leo. H. Koole
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Compositions of different molar ratios of polymethylmethacrylate-co-methacrylic acid (PMMA-co-MAA) were synthesized via free- radical polymerization. Polymer coated surfaces have been produced on silicon wafers. Coated samples were analyzed by atomic force microscopy (AFM). The results have shown that the roughness of the surfaces have increased by increasing the molar ratio of monomer methacrylic acid (MAA). This study reveals that the gradual increase in surface roughness is due to the fact that carboxylic functional groups have been generated by MAA segments. Such surfaces can be desirable platforms for fabrication of the biosensors for detection of the viruses and diseases.Keywords: polymethylmethacrylate-co-methacrylic acid (PMMA-co-MAA), polymeric material, atomic force microscopy, roughness, carboxylic functional groups
Procedia PDF Downloads 5955049 Robust Diagnosability of PEMFC Based on Bond Graph LFT
Authors: Ould Bouamama, M. Bressel, D. Hissel, M. Hilairet
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Fuel cell (FC) is one of the best alternatives of fossil energy. Recently, the research community of fuel cell has shown a considerable interest for diagnosis in view to ensure safety, security, and availability when faults occur in the process. The problematic for model based FC diagnosis consists in that the model is complex because of coupling of several kind of energies and the numerical values of parameters are not always known or are uncertain. The present paper deals with use of one tool: the Linear Fractional Transformation bond graph tool not only for uncertain modelling but also for monitorability (ability to detect and isolate faults) analysis and formal generation of robust fault indicators with respect to parameter uncertainties.The developed theory applied to a nonlinear FC system has proved its efficiency.Keywords: bond graph, fuel cell, fault detection and isolation (FDI), robust diagnosis, structural analysis
Procedia PDF Downloads 3665048 Impact of pH Control on Peptide Profile and Antigenicity of Whey Hydrolysates
Authors: Natalia Caldeira De Carvalho, Tassia Batista Pessato, Luis Gustavo R. Fernandes, Ricardo L. Zollner, Flavia Maria Netto
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Protein hydrolysates are ingredients of enteral diets and hypoallergenic formulas. Enzymatic hydrolysis is the most commonly used method for reducing the antigenicity of milk protein. The antigenicity and physicochemical characteristics of the protein hydrolysates depend on the reaction parameters. Among them, pH has been pointed out as of the major importance. Hydrolysis reaction in laboratory scale is commonly carried out under controlled pH (pH-stat). However, from the industrial point of view, controlling pH during hydrolysis reaction may be infeasible. This study evaluated the impact of pH control on the physicochemical properties and antigenicity of the hydrolysates of whey proteins with Alcalase. Whey protein isolate (WPI) solutions containing 3 and 7 % protein (w/v) were hydrolyzed with Alcalase 50 and 100 U g-1 protein at 60°C for 180 min. The reactions were carried out under controlled and uncontrolled pH conditions. Hydrolyses performed under controlled pH (pH-stat) were initially adjusted and maintained at pH 8.5. Hydrolyses carried out without pH control were initially adjusted to pH 8.5. Degree of hydrolysis (DH) was determined by OPA method, peptides profile was evaluated by HPLC-RP, and molecular mass distribution by SDS-PAGE/Tricine. The residual α-lactalbumin (α-La) and β-lactoglobulin (β-Lg) concentrations were determined using commercial ELISA kits. The specific IgE and IgG binding capacity of hydrolysates was evaluated by ELISA technique, using polyclonal antibodies obtained by immunization of female BALB/c mice with α-La, β-Lg and BSA. In hydrolysis under uncontrolled pH, the pH dropped from 8.5 to 7.0 during the first 15 min, remaining constant throughout the process. No significant difference was observed between the DH of the hydrolysates obtained under controlled and uncontrolled pH conditions. Although all hydrolysates showed hydrophilic character and low molecular mass peptides, hydrolysates obtained with and without pH control exhibited different chromatographic profiles. Hydrolysis under uncontrolled pH released, predominantly, peptides between 3.5 and 6.5 kDa, while hydrolysis under controlled pH released peptides smaller than 3.5 kDa. Hydrolysis with Alcalase under all conditions studied decreased by 99.9% the α-La and β-Lg concentrations in the hydrolysates detected by commercial kits. In general, β-Lg concentrations detected in the hydrolysates obtained under uncontrolled pH were significantly higher (p<0.05) than those detected in hydrolysates produced with pH control. The anti-α-La and anti-β-Lg IgE and IgG responses to all hydrolysates decreased significantly compared to WPI. Levels of specific IgE and IgG to the hydrolysates were below 25 and 12 ng ml-1, respectively. Despite the differences in peptide composition and α-La and β-Lg concentrations, no significant difference was found between IgE and IgG binding capacity of hydrolysates obtained with or without pH control. These results highlight the impact of pH on the hydrolysates characteristics and their concentrations of antigenic protein. Divergence between the antigen detection by commercial ELISA kits and specific IgE and IgG binding response was found in this study. This result shows that lower protein detection does not imply in lower protein antigenicity. Thus, the use of commercial kits for allergen contamination analysis should be cautious.Keywords: allergy, enzymatic hydrolysis, milk protein, pH conditions, physicochemical characteristics
Procedia PDF Downloads 3035047 Toxic Metal and Radiological Risk Assessment of Soil, Water and Vegetables around a Gold Mine Turned Residential Area in Mokuro Area of Ile-Ife, Osun State Nigeria: An Implications for Human Health
Authors: Grace O. Akinlade, Danjuma D. Maza, Oluwakemi O. Olawolu, Delight O. Babalola, John A. O. Oyekunle, Joshua O. Ojo
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The Mokuro area of Ile-Ife, South West Nigeria, was well known for gold mining in the past (about twenty years ago). However, the place has since been reclaimed and converted to residential area without any environmental risk assessment of the impact of the mining tailings on the environment. Soil, water, and plant samples were collected from 4 different locations around the mine-turned-residential area. Soil samples were pulverized and sieved into finer particles, while the plant samples were dried and pulverized. All the samples were digested and analyzed for As, Pb, Cd, and Zn using atomic absorption spectroscopy (AAS). From the analysis results, the hazard index (HI) was then calculated for the metals. The soil and plant samples were air dried and pulverized, then weighed, after which the samples were packed into special and properly sealed containers to prevent radon gas leakage. After the sealing, the samples were kept for 28 days to attain secular equilibrium. The concentrations of 40K, 238U, and 232Th in the samples were measured using a cesium iodide (CsI) spectrometer and URSA software. The AAS analysis showed that As, Pb, Cd (Toxic metals), and Zn (essential trace metals) are in concentrations lower than permissible limits in plants and soil samples, while the water samples had concentrations higher than permissible limits. The calculated health indices (HI) show that HI for water is >1 and that of plants and soil is <1. Gamma spectrometry result shows high levels of activity concentrations above the recommended limits for all the soil and plant samples collected from the area. Only the water samples have activity concentrations below the recommended limit. Consequently, the absorbed dose, annual effective dose, and excess lifetime cancer risk are all above the recommended safe limit for all the samples except for water samples. In conclusion, all the samples collected from the area are either contaminated with toxic metals or they pose radiological hazards to the consumers. Further detailed study is therefore recommended in order to be able to advise the residents appropriately.Keywords: toxic metals, gamma spectrometry, Ile-Ife, radiological hazards, gold mining
Procedia PDF Downloads 575046 Introducing the Concept of Sustainable Learning: Redesigning the Social Studies and Citizenship Education Curriculum in the Context of Saudi Arabia
Authors: Aiydh Aljeddani, Fran Martin
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Sustainable human development is an essential component of a sustainable economic, social and environmental development. Addressing sustainable learning only through the addition of new teaching methods, or embedding certain approaches, is not sufficient on its own to support the goals of sustainable human development. This research project seeks to explore how the process of redesigning the current principles of curriculum based on the concept of sustainable learning could contribute to preparing a citizen who could later contribute towards sustainable human development. Multiple qualitative methodologies were employed in order to achieve the aim of this study. The main research methods were teachers’ field notes, artefacts, informal interviews (unstructured interview), a passive participant observation, a mini nominal group technique (NGT), a weekly diary, and weekly meeting. The study revealed that the integration of a curriculum for sustainable development, in addition to the use of innovative teaching approaches, highly valued by students and teachers in social studies’ sessions. This was due to the fact that it created a positive atmosphere for interaction and aroused both teachers and students’ interest. The content of the new curriculum also contributed to increasing students’ sense of shared responsibility through involving them in thinking about solutions for some global issues. This was carried out through addressing these issues through the concept of sustainable development and the theory of Thinking Activity in a Social Context (TASC). Students had interacted with sustainable development sessions intellectually and they also practically applied it through designing projects and cut-outs. Ongoing meetings and workshops to develop work between both the researcher and the teachers, and by the teachers themselves, played a vital role in implementing the new curriculum. The participation of teachers in the development of the project through working papers, exchanging experiences and introducing amendments to the students' environment was also critical in the process of implementing the new curriculum. Finally, the concept of sustainable learning can contribute to the learning outcomes much better than the current curriculum and it can better develop the learning objectives in educational institutions.Keywords: redesigning, social studies and citizenship education curriculum, sustainable learning, thinking activity in a social context
Procedia PDF Downloads 2325045 Use of Didactic Bibliographic Resources to Improve the Teaching and Learning Processes of Animal Reproduction in Veterinary Science
Authors: Yasser Y. Lenis, Amy Jo Montgomery, Diego F. Carrillo-Gonzalez
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Introduction: The use of didactic instruments in different learning environments plays a pivotal role in enhancing the level of knowledge in veterinary science students. The direct instruction of basic animal reproduction concepts in students enrolled in veterinary medicine programs allows them to elucidate the biological and molecular mechanisms that perpetuate the animal species in an ecosystem. Therefore, universities must implement didactic strategies that facilitate the teaching and learning processes for students and, in turn, enrich learning environments. Objective: to evaluate the effect of the use of a didactic textbook on the level of theoretical knowledge in embryo-maternal recognition for veterinary medicine students. Methods: the participants (n=24) were divided into two experimental groups: control (Ctrl) and treatment (Treat). Both groups received 4 hours of theoretical training regarding the basic concepts in bovine embryo-maternal recognition. However, the Treat group was also exposed to a guided lecture and the activity play-to-learn from a cow reproduction didactic textbook. A pre-test and a post-test were applied to assess the prior and subsequent knowledge in the participants. Descriptive statistics were applied to identify the success rates for each of the tests. Afterwards, a repeated measures model was applied where the effect of the intervention was considered. Results: no significant difference (p>0,05) was observed in the number of right answers for groups Ctrl (54,2%±12,7) and Treat (40,8%±16,8) in the pre-test. There was no difference (p>0,05) compering the number of right answers in Ctrl pre-test (54,2%±12,7) and post-test (60,8±18,8). However, the Treat group showed a significant (p>0,05) difference in the number of right answers when comparing pre-test (40,8%±16,8) and post-test (71,7%±14,7). Finally, after the theoretical training and the didactic activity in the Treat group, an increase of 10.9% (p<0,05) in the number of right answers was found when compared with the Ctrl group. Conclusion: the use of didactic tools that include guided lectures and activities like play-to-learn from a didactic textbook enhances the level of knowledge in an animal reproduction course for veterinary medicine students.Keywords: animal reproduction, pedagogic, level of knowledge, learning environment
Procedia PDF Downloads 655044 An Approach Based on Statistics and Multi-Resolution Representation to Classify Mammograms
Authors: Nebi Gedik
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One of the significant and continual public health problems in the world is breast cancer. Early detection is very important to fight the disease, and mammography has been one of the most common and reliable methods to detect the disease in the early stages. However, it is a difficult task, and computer-aided diagnosis (CAD) systems are needed to assist radiologists in providing both accurate and uniform evaluation for mass in mammograms. In this study, a multiresolution statistical method to classify mammograms as normal and abnormal in digitized mammograms is used to construct a CAD system. The mammogram images are represented by wave atom transform, and this representation is made by certain groups of coefficients, independently. The CAD system is designed by calculating some statistical features using each group of coefficients. The classification is performed by using support vector machine (SVM).Keywords: wave atom transform, statistical features, multi-resolution representation, mammogram
Procedia PDF Downloads 2225043 Detecting Impact of Allowance Trading Behaviors on Distribution of NOx Emission Reductions under the Clean Air Interstate Rule
Authors: Yuanxiaoyue Yang
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Emissions trading, or ‘cap-and-trade', has been long promoted by economists as a more cost-effective pollution control approach than traditional performance standard approaches. While there is a large body of empirical evidence for the overall effectiveness of emissions trading, relatively little attention has been paid to other unintended consequences brought by emissions trading. One important consequence is that cap-and-trade could introduce the risk of creating high-level emission concentrations in areas where emitting facilities purchase a large number of emission allowances, which may cause an unequal distribution of environmental benefits. This study will contribute to the current environmental policy literature by linking trading activity with environmental injustice concerns and empirically analyzing the causal relationship between trading activity and emissions reduction under a cap-and-trade program for the first time. To investigate the potential environmental injustice concern in cap-and-trade, this paper uses a differences-in-differences (DID) with instrumental variable method to identify the causal effect of allowance trading behaviors on emission reduction levels under the clean air interstate rule (CAIR), a cap-and-trade program targeting on the power sector in the eastern US. The major data source is the facility-year level emissions and allowance transaction data collected from US EPA air market databases. While polluting facilities from CAIR are the treatment group under our DID identification, we use non-CAIR facilities from the Acid Rain Program - another NOx control program without a trading scheme – as the control group. To isolate the causal effects of trading behaviors on emissions reduction, we also use eligibility for CAIR participation as the instrumental variable. The DID results indicate that the CAIR program was able to reduce NOx emissions from affected facilities by about 10% more than facilities who did not participate in the CAIR program. Therefore, CAIR achieves excellent overall performance in emissions reduction. The IV regression results also indicate that compared with non-CAIR facilities, purchasing emission permits still decreases a CAIR participating facility’s emissions level significantly. This result implies that even buyers under the cap-and-trade program have achieved a great amount of emissions reduction. Therefore, we conclude little evidence of environmental injustice from the CAIR program.Keywords: air pollution, cap-and-trade, emissions trading, environmental justice
Procedia PDF Downloads 1515042 Hand Gestures Based Emotion Identification Using Flex Sensors
Authors: S. Ali, R. Yunus, A. Arif, Y. Ayaz, M. Baber Sial, R. Asif, N. Naseer, M. Jawad Khan
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In this study, we have proposed a gesture to emotion recognition method using flex sensors mounted on metacarpophalangeal joints. The flex sensors are fixed in a wearable glove. The data from the glove are sent to PC using Wi-Fi. Four gestures: finger pointing, thumbs up, fist open and fist close are performed by five subjects. Each gesture is categorized into sad, happy, and excited class based on the velocity and acceleration of the hand gesture. Seventeen inspectors observed the emotions and hand gestures of the five subjects. The emotional state based on the investigators assessment and acquired movement speed data is compared. Overall, we achieved 77% accurate results. Therefore, the proposed design can be used for emotional state detection applications.Keywords: emotion identification, emotion models, gesture recognition, user perception
Procedia PDF Downloads 2855041 Increased Risk of Adverse Birth Outcomes of Newborns in Arsenic Exposed- Women with Gestational Diabetes
Authors: Tania Mannan, Rahelee Zinnat, Fatema Jebunnesa, Israt Ara Hossain
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Background: Exposure to arsenic has known toxic effects but the effect on pregnancy outcomes is not as widely documented especially in women with diabetes. Growing evidence has suggested a potential role of arsenic exposure in the development of gestational diabetes mellitus (GDM). Therefore, we aimed to investigate the association of urinary arsenic (UAs) with birth outcomes in GDM subjects. Methods: Under an observational cross-sectional design a total of 263 GDM subjects (age in years, M±SD, 21±3.7) residing in an arsenic affected area of Bangladesh, were subjected to a 2 sample OGTT at the third trimester of gestation. Among them, 73 GDM and 190 non-GDM subjects enrolled in this study. Clinical and anthropometric measurements were done by standard techniques. Degree of chronic arsenic exposure was assessed by the level of UAs level. According to World Health Organization (WHO) criteria, GDM was diagnosed and neonatal outcomes using APGAR (Activity Pulse Grimace Appearance Respirations) Score, birth weight and size were assessed by a specialist obstetrician. Serum glucose was measured by the Glucose Oxidase method and UAs level was determined by ultraviolet/visible spectrophotometry. Result: Out of the 263 pregnant women, 28% developed GDM. Urinary Arsenic was significantly higher in the GDM as compared to the non-GDM group [UAs, µg/l, M±SD (range), 204.2±67.0 (67.0-377.0) vs 77.3±38.1 (22.0-99.0), p < 0.001]. Activity Pulse Grimace Appearance Respirations Score of the neonates from GDM mothers was significantly lower compared to the neonates from non-GDM mothers [APGAR Score, M±SD, 4.7±0.8 vs. 6.4±0.7, p<0.001]. Pearson’s correlation analysis in GDM subjects revealed that UA levels were found to have a significant positive correlation with both fasting and postprandial serum glucose levels (p < 0.001) and (p < 0.001) respectively. Again, a significant inverse correlation of UAs with birth weight and size was observed (p < 0.001). The APGAR Score of the neonates were found to have a significant negative correlation (p < 0.001) with UAs level. Conclusion: The effect of chronic arsenic exposure is associated with glucose intolerance during pregnancy and it also adversely affects birth outcomes. The study suggests further research on the impact of total arsenic exposure on pregnancy outcomes.Keywords: APGAR score, arsenic exposure, birth outcome, gestational diabetes mellitus,
Procedia PDF Downloads 1295040 Development of Antioxidant Rich Bakery Products by Applying Lysine and Maillard Reaction Products
Authors: Attila Kiss, Erzsébet Némedi, Zoltán Naár
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Due to the rapidly growing number of conscious customers in the recent years, more and more people look for products with positive physiological effects which may contribute to the preservation of their health. In response to these demands Food Science Research Institute of Budapest develops and introduces into the market new functional foods of guaranteed positive effect that contain bioactive agents. New, efficient technologies are also elaborated in order to preserve the maximum biological effect of the produced foods. The main objective of our work was the development of new functional biscuits fortified with physiologically beneficial ingredients. Bakery products constitute the base of the food nutrients’ pyramid, thus they might be regarded as foodstuffs of the largest consumed quantity. In addition to the well-known and certified physiological benefits of lysine, as an essential amino acid, a series of antioxidant type compounds is formed as a consequence of the occurring Maillard-reaction. Progress of the evoked Maillard-reaction was studied by applying diverse sugars (glucose, fructose, saccharose, isosugar) and lysine at several temperatures (120-170°C). Interval of thermal treatment was also varied (10-30 min). The composition and production technologies were tailored in order to reach the maximum of the possible biological benefits, so as to the highest antioxidant capacity in the biscuits. Out of the examined sugar components, theextent of the Maillard-reaction-driven transformation of glucose was the most pronounced at both applied temperatures. For the precise assessment of the antioxidant activity of the products FRAP and DPPH methods were adapted and optimised. To acquire an authentic and extensive mechanism of the occurring transformations, Maillard-reaction products were identified, and relevant reaction pathways were revealed. GC-MS and HPLC-MS techniques were applied for the analysis of the 60 generated MRPs and characterisation of actual transformation processes. 3 plausible major transformation routes might have been suggested based on the analytical result and the deductive sequence of possible occurring conversions between lysine and the sugars.Keywords: Maillard-reaction, lysine, antioxidant activity, GC-MS and HPLC-MS techniques
Procedia PDF Downloads 4825039 Artificial Neural Networks with Decision Trees for Diagnosis Issues
Authors: Y. Kourd, D. Lefebvre, N. Guersi
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This paper presents a new idea for fault detection and isolation (FDI) technique which is applied to industrial system. This technique is based on Neural Networks fault-free and Faulty behaviors Models (NNFM's). NNFM's are used for residual generation, while decision tree architecture is used for residual evaluation. The decision tree is realized with data collected from the NNFM’s outputs and is used to isolate detectable faults depending on computed threshold. Each part of the tree corresponds to specific residual. With the decision tree, it becomes possible to take the appropriate decision regarding the actual process behavior by evaluating few numbers of residuals. In comparison to usual systematic evaluation of all residuals, the proposed technique requires less computational effort and can be used for on line diagnosis. An application example is presented to illustrate and confirm the effectiveness and the accuracy of the proposed approach.Keywords: neural networks, decision trees, diagnosis, behaviors
Procedia PDF Downloads 5055038 Spatial Audio Player Using Musical Genre Classification
Authors: Jun-Yong Lee, Hyoung-Gook Kim
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In this paper, we propose a smart music player that combines the musical genre classification and the spatial audio processing. The musical genre is classified based on content analysis of the musical segment detected from the audio stream. In parallel with the classification, the spatial audio quality is achieved by adding an artificial reverberation in a virtual acoustic space to the input mono sound. Thereafter, the spatial sound is boosted with the given frequency gains based on the musical genre when played back. Experiments measured the accuracy of detecting the musical segment from the audio stream and its musical genre classification. A listening test was performed based on the virtual acoustic space based spatial audio processing.Keywords: automatic equalization, genre classification, music segment detection, spatial audio processing
Procedia PDF Downloads 4295037 Hard and Soft Skills in Marketing Education: Using Serious Games to Engage Higher Order Processing
Authors: Ann Devitt, Mairead Brady, Markus Lamest, Stephen Gomez
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This study set out to explore the use of an online collaborative serious game for student learning in a postgraduate introductory marketing module. The simulation game aimed to bridge the theory-practice divide in marketing by allowing students to apply theory in a safe, simulated marketplace. This study addresses the following research questions: Does an online marketing simulation game engage students higher order cognitive skills? Does collaborative activity required develop students’ “soft” skills, such as communication and negotiation? What specific affordances of the online simulation promote learning? This qualitative case study took place in 2014 with 40 postgraduate students on a Business Masters Programme. The two-week intensive module combined lectures with collaborative activity on a marketing simulation game, MMX from Pearsons. The game requires student teams to compete against other teams in a marketplace and design a marketing plan to maximize key performance indicators. The data for this study comprise essays written by students after the module reflecting on their learning on the module. A thematic analysis was conducted of the essays using the following a priori theme sets: 6 levels of the cognitive domain of Blooms taxonomy; 5 principles of Cooperative Learning; affordances of simulation environments including experiential learning; motivation and engagement; goal orientation. Preliminary findings would strongly suggest that the game facilitated students identifying the value of theory in practice, in particular for future employment; enhanced their understanding of group dynamics and their role within that; and impacted very strongly, both positively and negatively on motivation. In particular the game mechanics of MMX, which hinges on the correct identification of a target consumer group, was identified as a key determinant of extrinsic and intrinsic motivation for learners. The findings also suggest that the situation of the simulation game within a broader module which required post-game reflection was valuable in identifying key learning of marketing concepts in both the positive and the negative experiences of the game.Keywords: simulation, marketing, serious game, cooperative learning, bloom's taxonomy
Procedia PDF Downloads 5515036 Impacts of Artificial Intelligence on the Doctor-Patient Relationship: Ethical Principles, Informed Consent and Medical Obligation
Authors: Rafaella Nogaroli
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It is presented hypothetical cases in the context of AI algorithms to support clinical decisions, in order to discuss the importance of doctors to respect AI ethical principles. Regarding the principle of transparency and explanation, there is an impact on the new model of patient consent and on the understanding of qualified information. Besides, the human control of technology (AI as a tool) should guide the physician's activity; otherwise, he breaks the patient's legitimate expectation in a specific result, with the consequent transformation of the medical obligation nature.Keywords: medical law, artificial intelligence, ethical principles, patient´s informed consent, medical obligations
Procedia PDF Downloads 1025035 Mediterranean Diet, Duration of Admission and Mortality in Elderly, Hospitalized Patients: A Cross-Sectional Study
Authors: Christos Lampropoulos, Maria Konsta, Ifigenia Apostolou, Vicky Dradaki, Tamta Sirbilatze, Irini Dri, Christina Kordali, Vaggelis Lambas, Kostas Argyros, Georgios Mavras
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Objectives: Mediterranean diet has been associated with lower incidence of cardiovascular disease and cancer. The purpose of our study was to examine the hypothesis that Mediterranean diet may protect against mortality and reduce admission duration in elderly, hospitalized patients. Methods: Sample population included 150 patients (78 men, 72 women, mean age 80±8.2). The following data were taken into account in analysis: anthropometric and laboratory data, dietary habits (MedDiet score), patients’ nutritional status [Mini Nutritional Assessment (MNA) score], physical activity (International Physical Activity Questionnaires, IPAQ), smoking status, cause and duration of current admission, medical history (co-morbidities, previous admissions). Primary endpoints were mortality (from admission until 6 months afterwards) and duration of admission, compared to national guidelines for closed consolidated medical expenses. Logistic regression and linear regression analysis were performed in order to identify independent predictors for mortality and admission duration difference respectively. Results: According to MNA, nutrition was normal in 54/150 (36%) of patients, 46/150 (30.7%) of them were at risk of malnutrition and the rest 50/150 (33.3%) were malnourished. After performing multivariate logistic regression analysis we found that the odds of death decreased 30% per each unit increase of MedDiet score (OR=0.7, 95% CI:0.6-0.8, p < 0.0001). Patients with cancer-related admission were 37.7 times more likely to die, compared to those with infection (OR=37.7, 95% CI:4.4-325, p=0.001). According to multivariate linear regression analysis, admission duration was inversely related to Mediterranean diet, since it is decreased 0.18 days on average for each unit increase of MedDiet score (b:-0.18, 95% CI:-0.33 - -0.035, p=0.02). Additionally, the duration of current admission increased on average 0.83 days for each previous hospital admission (b:0.83, 95% CI:0.5-1.16, p<0.0001). The admission duration of patients with cancer was on average 4.5 days higher than the patients who admitted due to infection (b:4.5, 95% CI:0.9-8, p=0.015). Conclusion: Mediterranean diet adequately protects elderly, hospitalized patients against mortality and reduces the duration of hospitalization.Keywords: Mediterranean diet, malnutrition, nutritional status, prognostic factors for mortality
Procedia PDF Downloads 3135034 Evaluation of Arsenic Removal in Soils Contaminated by the Phytoremediation Technique
Authors: V. Ibujes, A. Guevara, P. Barreto
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Concentration of arsenic represents a serious threat to human health. It is a bioaccumulable toxic element and is transferred through the food chain. In Ecuador, values of 0.0423 mg/kg As are registered in potatoes of the skirts of the Tungurahua volcano. The increase of arsenic contamination in Ecuador is mainly due to mining activity, since the process of gold extraction generates toxic tailings with mercury. In the Province of Azuay, due to the mining activity, the soil reaches concentrations of 2,500 to 6,420 mg/kg As whereas in the province of Tungurahua it can be found arsenic concentrations of 6.9 to 198.7 mg/kg due to volcanic eruptions. Since the contamination by arsenic, the present investigation is directed to the remediation of the soils in the provinces of Azuay and Tungurahua by phytoremediation technique and the definition of a methodology of extraction by means of analysis of arsenic in the system soil-plant. The methodology consists in selection of two types of plants that have the best arsenic removal capacity in synthetic solutions 60 μM As, a lower percentage of mortality and hydroponics resistance. The arsenic concentrations in each plant were obtained from taking 10 ml aliquots and the subsequent analysis of the ICP-OES (inductively coupled plasma-optical emission spectrometry) equipment. Soils were contaminated with synthetic solutions of arsenic with the capillarity method to achieve arsenic concentration of 13 and 15 mg/kg. Subsequently, two types of plants were evaluated to reduce the concentration of arsenic in soils for 7 weeks. The global variance for soil types was obtained with the InfoStat program. To measure the changes in arsenic concentration in the soil-plant system, the Rhizo and Wenzel arsenic extraction methodology was used and subsequently analyzed with the ICP-OES (optima 8000 Pekin Elmer). As a result, the selected plants were bluegrass and llanten, due to the high percentages of arsenic removal of 55% and 67% and low mortality rates of 9% and 8% respectively. In conclusion, Azuay soil with an initial concentration of 13 mg/kg As reached the concentrations of 11.49 and 11.04 mg/kg As for bluegrass and llanten respectively, and for the initial concentration of 15 mg/kg As reached 11.79 and 11.10 mg/kg As for blue grass and llanten after 7 weeks. For the Tungurahua soil with an initial concentration of 13 mg/kg As it reached the concentrations of 11.56 and 12.16 mg/kg As for the bluegrass and llanten respectively, and for the initial concentration of 15 mg/kg As reached 11.97 and 12.27 mg/kg Ace for bluegrass and llanten after 7 weeks. The best arsenic extraction methodology of soil-plant system is Wenzel.Keywords: blue grass, llanten, phytoremediation, soil of Azuay, soil of Tungurahua, synthetic arsenic solution
Procedia PDF Downloads 1045033 Neuro-Preservation Potential of Resveratrol Against High Fat High Fructose-Induced Metabolic Syndrome
Authors: Rania F. Ahmed, Sally A. El Awdan, Gehad A. Abdel Jaleel, Dalia O. Saleh, Omar A. H. Ahmed-Farid
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The metabolic syndrome is an important public health concern often related to obesity, improper diet, and sedentary lifestyles and can predispose individuals to the development of many dangerous health conditions, disability and early death. This research aimed to investigate the efficacy of resveratrol (RSV) to reverse the neuro-complications associated with metabolic syndrome experimentally-induced in rats using an eight weeks high fat, high fructose diet (HFHF) model. The corresponding drug treatments were administered orally during the last 10 days of the diet. Behavioural tests namely the open field test (OFT) and the forced swimming test (FST) were conducted. Brain levels of monoamines viz. serotonin, norepinephrine and dopamine as well as their metabolites were assessed. 8-hydroxyguanosine (8-OHDG) as an indicative of DNA-fragmentation, nitric oxide (NOx) and tumor necrosis factor-α (TNF- α) were estimated. Finally, brain antioxidant parameters namely malondialdehyde (MDA), reduced and oxidized glutathione (GSH, GSSG) were evaluated. HFHF-induced metabolic syndrome resulted in decreased activity in the OFT and increased immobility duration in the FST. Furthermore, HFHF-induced metabolic syndrome lead to a significant increase in brain monoamines turn over as well as elevation in 8-OHDG, NOx, TNF- α, MDA and GSSG; and reduction in GSH. Ten days daily treatment with RSV (20 and 40 mg/kg p.o) dose dependently increased activity in the OFT and decreased immobility duration in the FST. Moreover, RSV normalized brain monoamines contents, reduced 8-OHDG, NOx, TNF- α, MDA and GSSG; and elevated GSH. In conclusion, we can say that RSV showed neuro-protective properties against HFHF-induced metabolic syndrome represented by monoamines preservation, prevention of neurodegeneration, anti-inflammatory and antioxidant potentials and could be recommended as a beneficial daily dietary supplement to treat the neuronal side effects associated with HFHF-induced metabolic syndrome.Keywords: antioxidants, DNA-fragmentation, forced swimming test, HFHF-induced metabolic syndrome, monoamines, nitric oxide (NOx), open field, resveratrol, tumor necrosis factor-α (TNF- α), 8-hydroxyguanosine (8-OHDG)
Procedia PDF Downloads 2765032 Use of Pragmatic Cues for Word Learning in Bilingual and Monolingual Children
Authors: Isabelle Lorge, Napoleon Katsos
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BACKGROUND: Children growing up in a multilingual environment face challenges related to the need to monitor the speaker’s linguistic abilities, more frequent communication failures, and having to acquire a large number of words in a limited amount of time compared to monolinguals. As a result, bilingual learners may develop different word learning strategies, rely more on some strategies than others, and engage cognitive resources such as theory of mind and attention skills in different ways. HYPOTHESIS: The goal of our study is to investigate whether multilingual exposure leads to improvements in the ability to use pragmatic inference for word learning, i.e., to use speaker cues to derive their referring intentions, often by overcoming lower level salience effects. The speaker cues we identified as relevant are (a) use of a modifier with or without stress (‘the WET dax’ prompting the choice of the referent which has a dry counterpart), (b) referent extension (‘this is a kitten with a fep’ prompting the choice of the unique rather than shared object), (c) referent novelty (choosing novel action rather than novel object which has been manipulated already), (d) teacher versus random sampling (assuming the choice of specific examples for a novel word to be relevant to the extension of that new category), and finally (e) emotional affect (‘look at the figoo’ uttered in a sad or happy voice) . METHOD: To this end, we implemented on a touchscreen computer a task corresponding to each of the cues above, where the child had to pick the referent of a novel word. These word learning tasks (a), (b), (c), (d) and (e) were adapted from previous word learning studies. 113 children have been tested (54 reception and 59 year 1, ranging from 4 to 6 years old) in a London primary school. Bilingual or monolingual status and other relevant information (age of onset, proficiency, literacy for bilinguals) is ascertained through language questionnaires from parents (34 out of 113 received to date). While we do not yet have the data that will allow us to test for effect of bilingualism, we can already see that performances are far from approaching ceiling in any of the tasks. In some cases the children’s performances radically differ from adults’ in a qualitative way, which means that there is scope for quantitative and qualitative effects to arise between language groups. The findings should contribute to explain the puzzling speed and efficiency that bilinguals demonstrate in acquiring competence in two languages.Keywords: bilingualism, pragmatics, word learning, attention
Procedia PDF Downloads 1395031 A Constructionist View of Projects, Social Media and Tacit Knowledge in a College Classroom: An Exploratory Study
Authors: John Zanetich
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Designing an educational activity that encourages inquiry and collaboration is key to engaging students in meaningful learning. Educational Information and Communications Technology (EICT) plays an important role in facilitating cooperative and collaborative learning in the classroom. The EICT also facilitates students’ learning and development of the critical thinking skills needed to solve real world problems. Projects and activities based on constructivism encourage students to embrace complexity as well as find relevance and joy in their learning. It also enhances the students’ capacity for creative and responsible real-world problem solving. Classroom activities based on constructivism offer students an opportunity to develop the higher–order-thinking skills of defining problems and identifying solutions. Participating in a classroom project is an activity for both acquiring experiential knowledge and applying new knowledge to practical situations. It also provides an opportunity for students to integrate new knowledge into a skill set using reflection. Classroom projects can be developed around a variety of learning objects including social media, knowledge management and learning communities. The construction of meaning through project-based learning is an approach that encourages interaction and problem-solving activities. Projects require active participation, collaboration and interaction to reach the agreed upon outcomes. Projects also serve to externalize the invisible cognitive and social processes taking place in the activity itself and in the student experience. This paper describes a classroom project designed to elicit interactions by helping students to unfreeze existing knowledge, to create new learning experiences, and then refreeze the new knowledge. Since constructivists believe that students construct their own meaning through active engagement and participation as well as interactions with others. knowledge management can be used to guide the exchange of both tacit and explicit knowledge in interpersonal interactions between students and guide the construction of meaning. This paper uses an action research approach to the development of a classroom project and describes the use of technology, social media and the active use of tacit knowledge in the college classroom. In this project, a closed group Facebook page becomes the virtual classroom where interaction is captured and measured using engagement analytics. In the virtual learning community, the principles of knowledge management are used to identify the process and components of the infrastructure of the learning process. The project identifies class member interests and measures student engagement in a learning community by analyzing regular posting on the Facebook page. These posts are used to foster and encourage interactions, reflect a student’s interest and serve as reaction points from which viewers of the post convert the explicit information in the post to implicit knowledge. The data was collected over an academic year and was provided, in part, by the Google analytic reports on Facebook and self-reports of posts by members. The results support the use of active tacit knowledge activities, knowledge management and social media to enhance the student learning experience and help create the knowledge that will be used by students to construct meaning.Keywords: constructivism, knowledge management, tacit knowledge, social media
Procedia PDF Downloads 2155030 Strategic Maintenance Management of Built Facilities in an Organisation
Authors: Anita D. Adamu, Winston M. W. Shakantu
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Maintenance management is no longer a stand-alone activity. It has now assumed a strategic position in many organisations that have recognised its importance in achieving primary goals and a key aspect of effective management of facilities. This paper aims at providing an understanding of the role and function of strategic management in creating and sustaining an effective maintenance management system in an organisation. The background provides an articulated concept and principles of strategic management. The theoretical concepts paved way for a conceptual framework for which strategic management can be integrated into the maintenance management system of an organisation to improve effectiveness in the maintenance of facilities.Keywords: facilities, maintenance management, organisations, strategic management
Procedia PDF Downloads 4095029 Presenting a Knowledge Mapping Model According to a Comparative Study on Applied Models and Approaches to Map Organizational Knowledge
Authors: Ahmad Aslizadeh, Farid Ghaderi
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Mapping organizational knowledge is an innovative concept and useful instrument of representation, capturing and visualization of implicit and explicit knowledge. There are a diversity of methods, instruments and techniques presented by different researchers following mapping organizational knowledge to reach determined goals. Implicating of these methods, it is necessary to know their exigencies and conditions in which those can be used. Integrating identified methods of knowledge mapping and comparing them would help knowledge managers to select the appropriate methods. This research conducted to presenting a model and framework to map organizational knowledge. At first, knowledge maps, their applications and necessity are introduced because of extracting comparative framework and detection of their structure. At the next step techniques of researchers such as Eppler, Kim, Egbu, Tandukar and Ebner as knowledge mapping models are presented and surveyed. Finally, they compare and a superior model would be introduced.Keywords: knowledge mapping, knowledge management, comparative study, business and management
Procedia PDF Downloads 4035028 High-Throughput Artificial Guide RNA Sequence Design for Type I, II and III CRISPR/Cas-Mediated Genome Editing
Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii
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A huge revolution has emerged in genome engineering by the discovery of CRISPR (clustered regularly interspaced palindromic repeats) and CRISPR-associated system genes (Cas) in bacteria. The function of type II Streptococcus pyogenes (Sp) CRISPR/Cas9 system has been confirmed in various species. Other S. thermophilus (St) CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been also reported for preventing phage infection. The CRISPR1-Cas system interferes by cleaving foreign dsDNA entering the cell in a length-specific and orientation-dependant manner. The S. thermophilus CRISPR3-Cas system also acts by cleaving phage dsDNA genomes at the same specific position inside the targeted protospacer as observed in the CRISPR1-Cas system. It is worth mentioning, for the effective DNA cleavage activity, RNA-guided Cas9 orthologs require their own specific PAM (protospacer adjacent motif) sequences. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of target site for the three orthogonals of Cas9 protein, a well-organized procedure will be required for high-throughput and accurate mining of possible target sites in a large genomic dataset. Consequently, we created a reliable procedure to explore potential gRNA sequences for type I (Streptococcus thermophiles), II (Streptococcus pyogenes), and III (Streptococcus thermophiles) CRISPR/Cas systems. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows: i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. The output of such procedure highlights the power of comparative genome mining for different CRISPR/Cas systems. This could yield a repertoire of Cas9 variants with expanded capabilities of gRNA design, and will pave the way for further advance genome and epigenome engineering.Keywords: CRISPR/Cas systems, gRNA mining, Streptococcus pyogenes, Streptococcus thermophiles
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