Search results for: Libor Matejka
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 19

Search results for: Libor Matejka

19 A Regional Analysis on Co-movement of Sovereign Credit Risk and Interbank Risks

Authors: Mehdi Janbaz

Abstract:

The global financial crisis and the credit crunch that followed magnified the importance of credit risk management and its crucial role in the stability of all financial sectors and the whole of the system. Many believe that risks faced by the sovereign sector are highly interconnected with banking risks and most likely to trigger and reinforce each other. This study aims to examine (1) the impact of banking and interbank risk factors on the sovereign credit risk of Eurozone, and (2) how the EU Credit Default Swaps spreads dynamics are affected by the Crude Oil price fluctuations. The hypothesizes are tested by employing fitting risk measures and through a four-staged linear modeling approach. The sovereign senior 5-year Credit Default Swap spreads are used as a core measure of the credit risk. The monthly time-series data of the variables used in the study are gathered from the DataStream database for a period of 2008-2019. First, a linear model test the impact of regional macroeconomic and market-based factors (STOXX, VSTOXX, Oil, Sovereign Debt, and Slope) on the CDS spreads dynamics. Second, the bank-specific factors, including LIBOR-OIS spread (the difference between the Euro 3-month LIBOR rate and Euro 3-month overnight index swap rates) and Euribor, are added to the most significant factors of the previous model. Third, the global financial factors including EURO to USD Foreign Exchange Volatility, TED spread (the difference between 3-month T-bill and the 3-month LIBOR rate based in US dollars), and Chicago Board Options Exchange (CBOE) Crude Oil Volatility Index are added to the major significant factors of the first two models. Finally, a model is generated by a combination of the major factor of each variable set in addition to the crisis dummy. The findings show that (1) the explanatory power of LIBOR-OIS on the sovereign CDS spread of Eurozone is very significant, and (2) there is a meaningful adverse co-movement between the Crude Oil price and CDS price of Eurozone. Surprisingly, adding TED spread (the difference between the three-month Treasury bill and the three-month LIBOR based in US dollars.) to the analysis and beside the LIBOR-OIS spread (the difference between the Euro 3M LIBOR and Euro 3M OIS) in third and fourth models has been increased the predicting power of LIBOR-OIS. Based on the results, LIBOR-OIS, Stoxx, TED spread, Slope, Oil price, OVX, FX volatility, and Euribor are the determinants of CDS spreads dynamics in Eurozone. Moreover, the positive impact of the crisis period on the creditworthiness of the Eurozone is meaningful.

Keywords: CDS, crude oil, interbank risk, LIBOR-OIS, OVX, sovereign credit risk, TED

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18 Utilization of Multi-Criteria Evaluation in Forensic Engineering and the Expertise outside Wall Subsystem

Authors: Tomas Barnak, Libor Matejka

Abstract:

The aim of this study is to create a standard application using multi-criteria evaluation in the field of forensic engineering. This situation can occur in the professional assessment in several cases such as when it is necessary to consider more criteria variant of the structural subsystems, more variants according to several criteria based on a court claim, which requires expert advice. A problematic situation arises when it is necessary to clearly determine the ranking of the options according to established criteria, and reduce subjective evaluation. For the procurement in the field of construction which is based on the prepared text of the law not only economic criteria but also technical, technological and environmental criteria will be determined. This fact substantially changes the style of evaluation of individual bids. For the above-mentioned needs of procurement, the unification of expert’s decisions and the use of multi-criteria assessment seem to be a reasonable option. In the case of experimental verification when using multi-criteria evaluation of alternatives construction subsystem the economic, technical, technological and environmental criteria will be compared. The core of the solution is to compare a selected number of set criteria, application methods and evaluation weighting based on the weighted values assigned to each of the criteria to use multi-criteria evaluation methods. The sequence of individual variations is determined by the evaluation of the importance of the values of corresponding criteria concerning expertise in the problematic of outside wall constructional subsystems.

Keywords: criteria, expertise, multi-criteria evaluation, outside wall subsystems

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17 Valuation of Caps and Floors in a LIBOR Market Model with Markov Jump Risks

Authors: Shih-Kuei Lin

Abstract:

The characterization of the arbitrage-free dynamics of interest rates is developed in this study under the presence of Markov jump risks, when the term structure of the interest rates is modeled through simple forward rates. We consider Markov jump risks by allowing randomness in jump sizes, independence between jump sizes and jump times. The Markov jump diffusion model is used to capture empirical phenomena and to accurately describe interest jump risks in a financial market. We derive the arbitrage-free model of simple forward rates under the spot measure. Moreover, the analytical pricing formulas for a cap and a floor are derived under the forward measure when the jump size follows a lognormal distribution. In our empirical analysis, we find that the LIBOR market model with Markov jump risk better accounts for changes from/to different states and different rates.

Keywords: arbitrage-free, cap and floor, Markov jump diffusion model, simple forward rate model, volatility smile, EM algorithm

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16 iPAD as a Communication Tool for Disabled Seniors: A Case Study

Authors: Vojtěch Gybas, Libor Klubal, Kateřina Kostolányová

Abstract:

This case study responds to the current trends in ICT. Mobile Touch iPads can provide very good assistance to disabled seniors. The intuitive tablet environment, the possibility of the formation environment and its portability, has a very positive effect on the use of particular communication. For comparison, using a conventional PC/notebook, word processor, keyboard and computer mouse compared to the iPad and selected applications. The results of this case study show that the use of mobile touch devices iPad for seniors with mental retardation is a great benefit. These devices do not require high demands on graphomotorics like a standard PC devices.

Keywords: ICT, iPad, handicapped seniors, communication, computer/notebook, applications, text editor

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15 Polyacrylate Modified Copper Nanoparticles with Controlled Size

Authors: Robert Prucek, Aleš Panáček, Jan Filip, Libor Kvítek, Radek Zbořil

Abstract:

The preparation of Cu nanoparticles (NPs) through the reduction of copper ions by sodium borohydride in the presence of sodium polyacrylate with a molecular weight of 1200 is reported. Cu NPs were synthesized at a concentration of copper salt equal to 2.5, 5, and 10 mM, and at a molar ratio of copper ions and monomeric unit of polyacrylate equal to 1:2. The as-prepared Cu NPs have diameters of about 2.5–3 nm for copper concentrations of 2.5 and 5 mM, and 6 nm for copper concentration of 10 mM. Depending on the copper salt concentration and concentration of additionally added polyacrylate to Cu particle dispersion, primarily formed NPs grow through the process of aggregation and/or coalescence into clusters and/or particles with a diameter between 20–100 nm. The amount of additionally added sodium polyacrylate influences the stability of Cu particles against air oxidation. The catalytic efficiency of the prepared Cu particles for the reduction of 4-nitrophenol is discussed.

Keywords: copper, nanoparticles, sodium polyacrylate, catalyst, 4-nitrophenol

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14 Modeling of Enthalpy and Heat Capacity of Phase-Change Materials

Authors: Igor Medved, Anton Trnik, Libor Vozar

Abstract:

Phase-change materials (PCMs) are of great interest in the applications where a temperature level needs to be maintained and/or where there is demand for thermal energy storage. Examples are storage of solar energy, cold, and space heating/cooling of buildings. During a phase change, the enthalpy vs. temperature plot of PCMs shows a jump and there is a distinct peak in the heat capacity plot. We present a theoretical description from which these jumps and peaks can be obtained. We apply our theoretical results to fit experimental data with very good accuracy for selected materials and changes between two phases. The development is based on the observation that PCMs are polycrystalline; i.e., composed of many single-crystalline grains. The enthalpy and heat capacity are thus interpreted as averages of the contributions from the individual grains. We also show how to determine the baseline and excess part of the heat capacity and thus the latent heat corresponding to the phase change.

Keywords: averaging, enthalpy jump, heat capacity peak, phase change

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13 Carbon Nitride Growth on ZnO Architectures for Enhanced Photoelectrochemical Water Splitting Application

Authors: Špela Hajduk, Sean P. Berglund, Matejka Podlogar, Goran Dražić, Fatwa F. Abdi, Zorica C. Orel, Menny Shalom

Abstract:

Graphitic carbon nitride materials (g-CN) have emerged as an attractive photocatalyst and electrocatalyst for photo and electrochemical water splitting reaction, due to their environmental benignity nature and suitable band gap. Many approaches were introduced to enhance the photoactivity and electronic properties of g-CN and resulted in significant changes in the electronic and catalytic properties. Here we demonstrate the synthesis of thin and homogenous g-CN layer on highly ordered ZnO nanowire (NW) substrate by growing a seeding layer of small supramolecular assemblies on the nanowires. The new synthetic approach leads to the formation of thin g-CN layer (~3 nm) without blocking all structure. Two different deposition methods of carbon nitride were investigated and will be presented. The amount of loaded carbon nitride significantly influences the PEC activity of hybrid material and all the ZnO/g-CNx electrodes show great improvement in photoactivity. The chemical structure, morphology and optical properties of the deposited g-CN were fully characterized by various techniques as X-ray powder spectroscopy (XRD), scanning electron microscopy (SEM), focused ion beam scanning electron microscopy (FIB-SEM), high-resolution scanning microscopy (HR-TEM) and X-ray photoelectron spectroscopy (XPS).

Keywords: carbon nitride, photoanode, solar water splitting, zinc oxide

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12 Non-Standard Monetary Policy Measures and Their Consequences

Authors: Aleksandra Nocoń (Szunke)

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The study is a review of the literature concerning the consequences of non-standard monetary policy, which are used by central banks during unconventional periods, threatening instability of the banking sector. In particular, the attention was paid to the effects of non-standard monetary policy tools for financial markets. However, the empirical evidence about their effects and real consequences for the financial markets are still not final. The main aim of the study is to survey the consequences of standard and non-standard monetary policy instruments, implemented during the global financial crisis in the United States, United Kingdom and Euroland, with particular attention to the results for the stabilization of global financial markets. The study analyses the consequences for short and long-term market interest rates, interbank interest rates and LIBOR-OIS spread. The study consists mainly of the empirical review, indicating the impact of the implementation of these tools for the financial markets. The following research methods were used in the study: literature studies, including domestic and foreign literature, cause and effect analysis and statistical analysis.

Keywords: asset purchase facility, consequences of monetary policy instruments, non-standard monetary policy, quantitative easing

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11 Synthesis of Flower-Like Silver Nanoarchitectures in Special Shapes and Their Applications in Surface-Enhanced Raman Scattering

Authors: Radka Králová, Libor Kvítek, Václav Ranc, Aleš Panáček, Radek Zbořil

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Surface–Enhanced Raman Scattering (SERS) is an optical spectroscopic technique with very good potential for sensitive detection of substances. In this research, active substrates with high enhancement were provided. Novel silver particles (nanostructures) with high roughened, flower–like morphology were prepared by reduction of cation complex [Ag(NH3)2]+ in presence of sodium borohydride as reducing agent and stabilized polyacrylic acid. The products were characterized by UV/VIS absorption spectrophotometry. Special shapes of silver particles were determined by scanning electron microscopy (SEM) and transmission electron spectroscopy (TEM). Dispersions of this particle were put on fixed substrate to producing suitable layer for SERS. Adenine was applied as basic substance whose effect of enhancement on the layer of silver nanostructures was studied. By comparison with our work, the important influence of stabilizers, polyacrylic acid with various molecular weight and concentration, on the transfer of particles and formation of new structure was confirmed.

Keywords: metals, nanostructures, chemical reduction, Raman spectroscopy, optical properties

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10 Body Composition Analyser Parameters and Their Comparison with Manual Measurements

Authors: I. Karagjozova, B. Dejanova, J. Pluncevic, S. Petrovska, V. Antevska, L. Todorovska

Abstract:

Introduction: Medical checking assessment is important in sports medicine. To follow the health condition in subjects who perform sports, body composition parameters, such as intracellular water, extracellular water, protein and mineral content, muscle and fat mass might be useful. The aim of the study was to show available parameters and to compare them to manual assessment. Material and methods: A number of 20 subjects (14 male and 6 female) at age of 20±2 years were determined in the study, 5 performed recreational sports, while others were professional ones. The mean height was 175±7 cm, the mean weight was 72±9 cm, and the body mass index (BMI) was 23±2 kg/m2. The measured compartments were as following: intracellular water (IW), extracellular water (EW), protein component (PC), mineral component (MC), skeletal muscle mass (SMM) and body fat mass (BFM). Lean balance were examined for right and left arm (LA), trunk (T), right leg (RL) and left leg (LL). The comparison was made between the calculation derived by manual made measurements, using Matejka formula and parameters obtained by body composition analyzer (BCA) - Inbody 720 BCA Biospace. Used parameters for the comparison were muscle mass (SMM), body fat mass (BFM). Results: BCA obtained values were for: IW - 22.6±5L, EW - 13.5±2 L, PC - 9.8±0.9 kg, MC - 3.5±0.3, SMM - 27±3 kg, BFM - 13.8±4 kg. Lean balance showed following values for: RA - 2.45±0.2 kg, LA - 2.37±0.4, T - 20.9±5 kg, RL - 7.43±1 kg, and LL - 7.49 ±1.5 kg. SMM showed statistical difference between manual obtained value, 51±01% to BCA parameter 45.5±3% (p<0.001). Manual obtained values for BFM was lower (17±2%) than BCA obtained one, 19.5±5.9% (p<0.02). Discussion: The obtained results showed appropriate values for the examined age, regarding to all examined parameters which contribute to overview the body compartments, important for sport performing. Due to comparison between the manual and BCA assessment, we may conclude that manual measurements may differ from the certain ones, which is confirmed by statistical significance.

Keywords: athletes, body composition, bio electrical impedance, sports medicine

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9 System for Mechanical Stimulation of the Mesenchymal Stem Cells Supporting Differentiation into Osteogenic Cells

Authors: Jana Stepanovska, Roman Matejka, Jozef Rosina, Marta Vandrovcova, Lucie Bacakova

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The aim of this study was to develop a system for mechanical and also electrical stimulation controlling in vitro osteogenesis under conditions more similar to the in vivo bone microenvironment than traditional static cultivation, which would achieve good adhesion, growth and other specific behaviors of osteogenic cells in cultures. An engineered culture system for mechanical stimulation of the mesenchymal stem cells on the charged surface was designed. The bioreactor allows efficient mechanical loading inducing an electrical response and perfusion of the culture chamber with seeded cells. The mesenchymal stem cells were seeded to specific charged materials, like polarized hydroxyapatite (Hap) or other materials with piezoelectric and ferroelectric features, to create electrical potentials for stimulating of the cells. The material of the matrix was TiNb alloy designed for these purposes, and it was covered by BaTiO3 film, like a kind of piezoelectric material. The process of mechanical stimulation inducing electrical response is controlled by measuring electrical potential in the chamber. It was performed a series of experiments, where the cells were seeded, perfused and stimulated up to 48 hours under different conditions, especially pressure and perfusion. The analysis of the proteins expression was done, which demonstrated the effective mechanical and electrical stimulation. The experiments demonstrated effective stimulation of the cells in comparison with the static culture. This work was supported by the Ministry of Health, grant No. 15-29153A and the Grant Agency of the Czech Republic grant No. GA15-01558S.

Keywords: charged surface, dynamic cultivation, electrical stimulation, ferroelectric layers, mechanical stimulation, piezoelectric layers

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8 Assisted Approach as a Tool for Increasing Attention When Using the iPad in a Special Elementary School: Action Research

Authors: Vojtěch Gybas, Libor Klubal, Kateřina Kostolányová

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Nowadays, mobile touch technologies, such as tablets, are an integral part of teaching and learning in many special elementary schools. Many special education teachers tend to choose an iPad tablet with iOS. The reason is simple; the iPad has a function for pupils with special educational needs. If we decide to use tablets in teaching, in general, first we should try to stimulate the cognitive abilities of the pupil at the highest level, while holding the pupil’s attention on the task, when working with the device. This paper will describe how student attention can be increased by eliminating the working environment of selected applications, while using iPads with pupils in a special elementary school. Assisted function approach is highly effective at eliminating unwanted touching by a pupil when working on the desktop iPad, thus actively increasing the pupil´s attention while working on specific educational applications. During the various stages of the action, the research was conducted via data collection and interpretation. After a phase of gaining results and ideas for practice and actions, we carried out the check measurement, this time using the tool-assisted approach. In both cases, the pupils worked in the Math Board application and the resulting differences were evident.

Keywords: special elementary school, a mobile touch device, iPad, attention, Math Board

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7 Individualized Teaching Process for Pupils with Moderate Mental Disability

Authors: VojtěCh Gybas, Libor Klubal, KateřIna KostoláNyová

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Individualized teaching process for pupils with moderate mental disabilities with the help of using mobile touch devices may be one of the forms of teaching to achieve better development of these students during the teaching process. Didactics of information and communication technology (ICT) for special primary schools, where within the Czech Republic pupils with moderate mental retardation are educated, is not precisely and clearly defined. Still, general educational program for elementary school contains a special educational area of information and communication technology, in which the work and content area are focused on work with the classic desktop, and it is not always acceptable in the case of students with moderate mental disabilities. Individualization of their schooling requires a fully elaborate content of teaching material corresponding with intellectual abilities and individuality of each pupil. After three years of daily use of mobile touch devices iPad and participant observation of 7 pupils in a class from special elementary school, we can say that these technologies can be a very useful tool, and in many ways, they even exceed, compensate and replace freely available printed educational material that is rather outdated. By working with mobile touch technology, a pupil gains responsibility, trains his will, learns to rely on himself. The first results obtained from the case studies suggest that this form of teaching may also be beneficial for pupils with moderate mental disabilities.

Keywords: individualized teaching, mobile touch technology, iPad, moderate mental disability, special education needs

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6 Rate of Profit as a Pricing Benchmark in Islamic Banking to Create Financial Stability

Authors: Trisiladi Supriyanto

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Although much research has been done on the pricing benchmark both in terms of fiqh or Islamic economic perspective, but no substitution for the concept of interest (rate of interest) up to now in the application of Islamic Banking because some of the jurists from the middle east even allow the use of a benchmark rate such as LIBOR (London Interbank Offered Rate) as a measure of Islamic financial asset prices, so in other words, they equate the concept of rate of interest with the concept of rate of profit, which is the core reason (raison detre) for the replacement of usury as instructed in the Quran. This study aims to find the concept of rate of profit on Islamic banking that can create economic justice and stability in Islamic Banking and Capital market. Rate of profit that creates economic justice and stability can be achieved through its role in maintaining the stability of the financial system in which there is an equitable distribution of income and wealth. To determine the role of the rate of profit as the basis of the sharing system implemented in the Islamic financial system, we can see the connection of rate of profit in creating financial stability, especially in the asset-liability management of financial institutions that generate a stable net margin or the rate of profit that is not affected by the ups and downs of the market risk factors including indirect effect on interest rates. Furthermore, Islamic financial stability can be seen from the role of the rate of profit on the stability of the Islamic financial assets that are measured from the Islamic financial asset price volatility in Islamic Bond Market in Capital Market.

Keywords: Rate of profit, economic justice, stability, equitable distribution of income, equitable distribution of wealth

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5 Effects of Drought on Microbial Activity in Rhizosphere, Soil Hydrophobicity and Leaching of Mineral Nitrogen from Arable Soil Depending on Method of Fertilization

Authors: Jakub Elbl, Lukáš Plošek, Antonín Kintl, Jaroslav Hynšt, Soňa Javoreková, Jaroslav Záhora, Libor Kalhotka, Olga Urbánková, Ivana Charousová

Abstract:

This work presents the first results from the long-term laboratory experiment dealing with impact of drought on soil properties. Three groups of the treatment (A, B and C) with different regime of irrigation were prepared. The soil water content was maintained at 70 % of soil water holding capacity in group A, at 40 % in group B. In group C, soil water regime was maintained in the range of wilting point. Each group of the experiment was divided into three variants (A1 = B1, C1; A2 = B2, C2 etc.) with three repetitions: Variants A1 (B1, C1) were controls without addition of another fertilizer. Variants A2 (B2, C2) were fertilized with mineral nitrogen fertilizer DAM 390 (0.140 Mg of N per ha) and variants A3 (B3, C3) contained 45 g of Cp per a pot. The significant differences (ANOVA, P<0.05) in the leaching of mineral nitrogen and values of saturated hydraulic conductivity (Ksat) were found. The highest values of Ksat were found in variants (within each group) with addition of compost (A3, B3, C3). Conversely, the lowest values of Ksat were found in variants with addition of mineral nitrogen. Low values of Ksat indicate an increased level of hydrophobicity in individual groups of the experiment. Moreover, all variants with compost addition showed lower amount of mineral nitrogen leaching and high level of microbial activity than variants without. This decrease of mineral nitrogen leaching was about 200 % in comparison with the control variant and about 300 % with variant, where mineral nitrogen was added. Based on these results, we can conclude that changes of soil water content directly have impact on microbial activity, soil hydrophobicity and loss of mineral nitrogen from the soil.

Keywords: drought, microbial activity, mineral nitrogen, soil hydrophobicity

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4 Bioreactor for Cell-Based Impedance Measuring with Diamond Coated Gold Interdigitated Electrodes

Authors: Roman Matejka, Vaclav Prochazka, Tibor Izak, Jana Stepanovska, Martina Travnickova, Alexander Kromka

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Cell-based impedance spectroscopy is suitable method for electrical monitoring of cell activity especially on substrates that cannot be easily inspected by optical microscope (without fluorescent markers) like decellularized tissues, nano-fibrous scaffold etc. Special sensor for this measurement was developed. This sensor consists of corning glass substrate with gold interdigitated electrodes covered with diamond layer. This diamond layer provides biocompatible non-conductive surface for cells. Also, a special PPFC flow cultivation chamber was developed. This chamber is able to fix sensor in place. The spring contacts are connecting sensor pads with external measuring device. Construction allows real-time live cell imaging. Combining with perfusion system allows medium circulation and generating shear stress stimulation. Experimental evaluation consist of several setups, including pure sensor without any coating and also collagen and fibrin coating was done. The Adipose derived stem cells (ASC) and Human umbilical vein endothelial cells (HUVEC) were seeded onto sensor in cultivation chamber. Then the chamber was installed into microscope system for live-cell imaging. The impedance measurement was utilized by vector impedance analyzer. The measured range was from 10 Hz to 40 kHz. These impedance measurements were correlated with live-cell microscopic imaging and immunofluorescent staining. Data analysis of measured signals showed response to cell adhesion of substrates, their proliferation and also change after shear stress stimulation which are important parameters during cultivation. Further experiments plan to use decellularized tissue as scaffold fixed on sensor. This kind of impedance sensor can provide feedback about cell culture conditions on opaque surfaces and scaffolds that can be used in tissue engineering in development artificial prostheses. This work was supported by the Ministry of Health, grants No. 15-29153A and 15-33018A.

Keywords: bio-impedance measuring, bioreactor, cell cultivation, diamond layer, gold interdigitated electrodes, tissue engineering

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3 Leveraging xAPI in a Corporate e-Learning Environment to Facilitate the Tracking, Modelling, and Predictive Analysis of Learner Behaviour

Authors: Libor Zachoval, Daire O Broin, Oisin Cawley

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E-learning platforms, such as Blackboard have two major shortcomings: limited data capture as a result of the limitations of SCORM (Shareable Content Object Reference Model), and lack of incorporation of Artificial Intelligence (AI) and machine learning algorithms which could lead to better course adaptations. With the recent development of Experience Application Programming Interface (xAPI), a large amount of additional types of data can be captured and that opens a window of possibilities from which online education can benefit. In a corporate setting, where companies invest billions on the learning and development of their employees, some learner behaviours can be troublesome for they can hinder the knowledge development of a learner. Behaviours that hinder the knowledge development also raise ambiguity about learner’s knowledge mastery, specifically those related to gaming the system. Furthermore, a company receives little benefit from their investment if employees are passing courses without possessing the required knowledge and potential compliance risks may arise. Using xAPI and rules derived from a state-of-the-art review, we identified three learner behaviours, primarily related to guessing, in a corporate compliance course. The identified behaviours are: trying each option for a question, specifically for multiple-choice questions; selecting a single option for all the questions on the test; and continuously repeating tests upon failing as opposed to going over the learning material. These behaviours were detected on learners who repeated the test at least 4 times before passing the course. These findings suggest that gauging the mastery of a learner from multiple-choice questions test scores alone is a naive approach. Thus, next steps will consider the incorporation of additional data points, knowledge estimation models to model knowledge mastery of a learner more accurately, and analysis of the data for correlations between knowledge development and identified learner behaviours. Additional work could explore how learner behaviours could be utilised to make changes to a course. For example, course content may require modifications (certain sections of learning material may be shown to not be helpful to many learners to master the learning outcomes aimed at) or course design (such as the type and duration of feedback).

Keywords: artificial intelligence, corporate e-learning environment, knowledge maintenance, xAPI

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2 Mesenchymal Stem Cells on Fibrin Assemblies with Growth Factors

Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

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Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

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1 Surface Adjustments for Endothelialization of Decellularized Porcine Pericardium

Authors: M. Markova, E. Filova, O. Kaplan, R. Matejka, L. Bacakova

Abstract:

The porcine pericardium is used as a material for cardiac and aortic valves substitutes. Current biological aortic heart valve prosthesis have a limited lifetime period because they undergo degeneration. In order to make them more biocompatible and prolong their lifetime it is necessary to reseed the decellularized prostheses with endothelial cells and with valve interstitial cells. The endothelialization of the prosthesis-surface may be supported by suitable chemical surface modification of the prosthesis. The aim of this study is to prepare bioactive fibrin layers which would both support endothelialization of porcine pericardium and enhance differentiation and maturation of the endothelial cells seeded. As a material for surface adjustments we used layers of fibrin with/without heparin and some of them with adsorbed or chemically bound FGF2, VEGF or their combination. Fibrin assemblies were prepared in 24-well cell culture plate and were seeded with HSVEC (Human Saphenous Vein Endothelial Cells) at a density of 20,000 cells per well in EGM-2 medium with 0.5% FS and without heparin, without FGF2 and without VEGF; medium was supplemented with aprotinin (200 U/mL). As a control, surface polystyrene (PS) was used. Fibrin was also used as homogeneous impregnation of the decellularized porcine pericardium throughout the scaffolds. Morphology, density, and viability of the seeded endothelial cells were observed from micrographs after staining the samples by LIVE/DEAD cytotoxicity/viability assay kit on the days 1, 3, and 7. Endothelial cells were immunocytochemically stained for proteins involved in cell adhesion, i.e. alphaV integrin, vinculin, and VE-cadherin, markers of endothelial cells differentiation and maturation, i.e. von Willebrand factor and CD31, and for extracellular matrix proteins typically produced by endothelial cells, i.e. type IV collagen and laminin. The staining intensities were subsequently quantified using a software. HSVEC cells grew on each of the prepared surfaces better than on control surface. They reached confluency. The highest cell densities were obtained on the surface of fibrin with heparin and both grow factors used together. Intensity of alphaV integrins staining was highest on samples with remained fibrin layer, i.e. on layers with lower cell densities, i.e. on fibrin without heparin. Vinculin staining was apparent, but was rather diffuse, on fibrin with both FGF2 and VEGF and on control PS. Endothelial cells on all samples were positively stained for von Willebrand factor and CD31. VE-cadherin receptors clusters were best developed on fibrin with heparin and growth factors. Significantly stronger staining of type IV collagen was observed on fibrin with heparin and both growth factors. Endothelial cells on all samples produced laminin-1. Decellularized pericardium was homogeneously filled with fibrin structures. These fibrin-modified pericardium samples will be further seeded with cells and cultured in a bioreactor. Fibrin layers with/without heparin and with adsorbed or chemically bound FGF2, VEGF or their combination are good surfaces for endothelialization of cardiovascular prostheses or porcine pericardium based heart valves. Supported by the Ministry of Health, grants No15-29153A and 15-32497A, and the Grant Agency of the Czech Republic, project No. P108/12/G108.

Keywords: aortic valves prosthesis, FGF2, heparin, HSVEC cells, VEGF

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