Search results for: Katarina Holla
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 30

Search results for: Katarina Holla

30 Evaluation of European Surveys in the Area of Health and Safety at Work and Identification of New Risks in the Labor Environment

Authors: Alena Dadova, Katarina Holla, Anna Cidlinova, Linda Makovicka Osvaldova, Jiri Vala, Samuel Kockar

Abstract:

Occupational health and safety (ASH) is an area in which procedures and applications are constantly evolving and changing through legislation and new directives and guidelines. In this way, the relevant organizations strive to ensure continuous progress and the advantage of up-to-date information to ensure safety and prevent occupational accidents. Three ESENER surveys have been carried out in the European Union, led by the Agency for Safety and Health at Work (EU-OSHA). On the basis of surveys, it was determined how European workplaces manage risks and how they manage the field of safety and health protection at work. Thousands of companies and organizations in the European Union were involved in the surveys. Organizations and businesses were presented with a questionnaire that focused on the following topics: the impact of general risks on the field of OSH and the possibility of their management, psychosocial risks and other factors such as stress, harassment and bullying, and employee participation in OSH procedures. The article is dedicated to the fundamental conclusions from these surveys and their subsequent connection with the strategic intent of the Strategic Framework of European Union for the years 2021 - 2027. In the conclusion, emerging risks are identified and EU will soon have to deal with them.

Keywords: ESENER, emerging risks, strategic framework in OSH, EU

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29 Importance of Risk Assessment in Managers´ Decision-Making Process

Authors: Mária Hudáková, Vladimír Míka, Katarína Hollá

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Making decisions is the core of management and a result of conscious activities which is under way in a particular environment and concrete conditions. The managers decide about the goals, procedures and about the methods how to respond to the changes and to the problems which developed. Their decisions affect the effectiveness, quality, economy and the overall successfulness in every organisation. In spite of this fact, they do not pay sufficient attention to the individual steps of the decision-making process. They emphasise more how to cope with the individual methods and techniques of making decisions and forget about the way how to cope with analysing the problem or assessing the individual solution variants. In many cases, the underestimating of the analytical phase can lead to an incorrect assessment of the problem and this can then negatively influence its further solution. Based on our analysis of the theoretical solutions by individual authors who are dealing with this area and the realised research in Slovakia and also abroad we can recognise an insufficient interest of the managers to assess the risks in the decision-making process. The goal of this paper is to assess the risks in the managers´ decision-making process relating to the conditions of the environment, to the subject’s activity (the manager’s personality), to the insufficient assessment of individual variants for solving the problems but also to situations when the arisen problem is not solved. The benefit of this paper is the effort to increase the need of the managers to deal with the risks during the decision-making process. It is important for every manager to assess the risks in his/her decision-making process and to make efforts to take such decisions which reflect the basic conditions, states and development of the environment in the best way and especially for the managers´ decisions to contribute to achieving the determined goals of the organisation as effectively as possible.

Keywords: risk, decision-making, manager, process, analysis, source of risk

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28 Calculate Consumer Surplus and Producer Surplus Using Integration

Authors: Bojan Radisic, Katarina Stavlic

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The paper describes two economics terms consumer surplus and producer surplus using the definite integrals (the Riemann integral). The consumer surplus is the difference between what consumers are willing to pay and actual price. The producer surplus is the difference between what producers selling at the current price, rather than at the price they would have been are willing to accept. Using the definite integrals describe terms and mathematical formulas of the consumer surplus and the producer surplus and will be applied to the numerical examples.

Keywords: consumer surplus, producer surplus, definite integral, integration

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27 Perceptions of Doctors and Nurses About Euthanasia in Indian Scenario

Authors: B. Unnikrishnan, Tanuj Kanchan, Ramesh Holla, Nithin Kumar

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Euthanasia has been debated for the ethical, legal, social, and religious implications associated with it. The present research was conducted to study the perceptions of doctors and nurses about ethical and legal aspects of Euthanasia in Indian scenario. The study was carried out at three tertiary care hospitals of Kasturba Medical College (KMC), Mangalore, India. Practicing doctors and nurses working in the hospitals associated with KMC were included in the study after taking written informed consent from the participants. The data was analyzed using SPSS version 11.5. Mann-Whitney U test was used to compare the responses of doctors and nurses. P-value of <0.05 was taken as statistically significant. A total of 144 doctors and nurses participated in the study. Both doctors and nurses agreed that if a terminally ill patient wishes to die, the wish cannot be honored ethically and legally. A significantly larger number of nurses agreed that patient’s wish for euthanasia cannot be honored ethically and legally when compared to the doctors. Though the doctors and nurses were broadly in agreement with the existing legal and ethical views on the issue, their knowledge on the issue with regard to the legal status of euthanasia in India and ethical aspects relating to it needs to be strengthened.

Keywords: euthanasia, ethical aspects, legal aspects, India

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26 Trace Metals in Natural Bottled Water on Montenegrin Market and Comaparison with Tap Water in Podgorica

Authors: Katarina Živković, Ivana Joksimović

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Many different chemicals may occur in drinking water and cause significant human health risks after prolonged periods of exposure. In particular concern are contaminants that have cumulative toxic properties, such as heavy metals. This investigation was done to clarify concerns about chemical quality and safety of drinking tap water in Podgorica. For comparison, all available natural bottled water on Montenegrin market were bought. All samples (bottled water and tap water from Podgorica) were analyzed using ICP –OES on contents of Al, Cd, Pb, Cu, Zn,Cr, Fe, As and Mn. All results compared with the maximum concentration levels allowed by international standards and World Health Organization (WHO) guidelines. The results of analysis showed that all trace of heavy metals were very low and in same time below MCL according to WHO and International standard.

Keywords: inductively coupled plasma - optical emission spectrometry (ICP-OES), Montenegro (Podgorica), natural bottled water, tap water , trace of heavy metal

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25 Traffic Signal Control Using Citizens’ Knowledge through the Wisdom of the Crowd

Authors: Aleksandar Jovanovic, Katarina Kukic, Ana Uzelac, Dusan Teodorovic

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Wisdom of the Crowd (WoC) is a decentralized method that uses the collective intelligence of humans. Individual guesses may be far from the target, but when considered as a group, they converge on optimal solutions for a given problem. We will utilize WoC to address the challenge of controlling traffic lights within intersections from the streets of Kragujevac, Serbia. The problem at hand falls within the category of NP-hard problems. We will employ an algorithm that leverages the swarm intelligence of bees: Bee Colony Optimization (BCO). Data regarding traffic signal timing at a single intersection will be gathered from citizens through a survey. Results obtained in that manner will be compared to the BCO results for different traffic scenarios. We will use Vissim traffic simulation software as a tool to compare the performance of bees’ and humans’ collective intelligence.

Keywords: wisdom of the crowd, traffic signal control, combinatorial optimization, bee colony optimization

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24 Objective and Subjective Preconditions for Entrepreneurship: From the Point of View of Enterprise Risk Management

Authors: Maria Luskova, Maria Hudakova, Katarina Buganova

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Established objective and subjective preconditions for entrepreneurship, forming the business organically related whole, are the necessary condition of successful entrepreneurial activities. Objective preconditions for entrepreneurship are developed by the market economy that should stimulate entrepreneurship by allowing the use of economic opportunities for all those who want to do business in respective field while providing guarantees to all owners and creating a stable business environment for entrepreneurs. Subjective preconditions of entrepreneurship are formed primarily by personal characteristics of the entrepreneur. These are his properties, abilities, skills, physiological, and psychological preconditions which may be inherited, inborn or sequentially developed and obtained during his life on the basis of education and influences of surrounding environment. The paper is dealing with issues of objective and subjective preconditions for entrepreneurship and provides their analysis in view of the current situation in Slovakia. It presents risks of the business environment in Slovakia that the Slovak managers considered the most significant in 2014 and defines the dominant attributes of the entrepreneur in the current business environment in Slovakia.

Keywords: entrepreneurship, innovations, opportunity, risk, uncertainty

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23 New Possibilities for Testing UX and UI Design on Mobile Devices

Authors: Jakub Berčík, Anna Mravcová, Jana Gálová, Katarína Neomániová

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In an era when everything is increasingly digital, consumers are always looking for new options in solutions to their everyday needs. In this context, mobile apps are developing at an exponential pace. One of the fastest growing segments of mobile technologies is, obviously, e-commerce. It can be predicted that mobile commerce will record nearly three times the global growth of e-commerce across all platforms, which indicates its importance in the given segment. The current coronavirus pandemic is also changing many of the existing paradigms both socially, economically, and technologically, which has a major impact on changing consumer behaviour and the emphasis on simplification and clarity of mobile solutions. This is the area that user experience (UX) and user interface (UI) designers deal with. Their task is to design a sufficiently attractive and interesting solution that will be available on all mobile devices and at the same time will be easy enough for the customer/visitor to get to the destination or to get the necessary information in a few clicks. The basis for changes in UX design can now be obtained not only through online analytical tools but also through neuromarketing, especially in the case of mobile devices. The paper highlights new possibilities for testing UX design applications on mobile devices using a special platform that combines a stationary eye camera (eye tracking) and facial analysis (facial coding).

Keywords: emotions, mobile design, user experience, visual attention

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22 Machine Learning Methods for Flood Hazard Mapping

Authors: Stefano Zappacosta, Cristiano Bove, Maria Carmela Marinelli, Paola di Lauro, Katarina Spasenovic, Lorenzo Ostano, Giuseppe Aiello, Marco Pietrosanto

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This paper proposes a novel neural network approach for assessing flood hazard mapping. The core of the model is a machine learning component fed by frequency ratios, namely statistical correlations between flood event occurrences and a selected number of topographic properties. The proposed hybrid model can be used to classify four different increasing levels of hazard. The classification capability was compared with the flood hazard mapping River Basin Plans (PAI) designed by the Italian Institute for Environmental Research and Defence, ISPRA (Istituto Superiore per la Protezione e la Ricerca Ambientale). The study area of Piemonte, an Italian region, has been considered without loss of generality. The frequency ratios may be used as a standalone block to model the flood hazard mapping. Nevertheless, the mixture with a neural network improves the classification power of several percentage points, and may be proposed as a basic tool to model the flood hazard map in a wider scope.

Keywords: flood modeling, hazard map, neural networks, hydrogeological risk, flood risk assessment

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21 Chromatographic Lipophilicity Determination of Newly Synthesized Steroid Derivatives for Further Biological Analysis

Authors: Milica Z. Karadzic, Lidija R. Jevric, Sanja Podunavac-Kuzmanovic, Strahinja Z. Kovacevic, Anamarija I. Mandic, Katarina Penov-Gasi, Andrea R. Nikolic, Aleksandar M. Okljesa

Abstract:

In this study, a set of 29 newly synthesized steroid derivatives were investigated using reversed-phase high-performance liquid chromatography (RP-HPLC) as a first step in preselection of drug candidates. This analysis presents an experimental determination of chromatographic lipophilicity, and it was conducted to obtain physicochemical characterization of these molecules. As the most widely used bonded phases in RP-HPLC, octadecyl (C18) and octyl (C8) were used. Binary mixtures of water and acetonitrile or methanol were used as mobile phases. Obtained results were expressed as retention factor values logk and they were correlated with logP values. The results showed that both columns provide good estimations of the chromatographic lipophilicity of the molecules included in this study. This analysis was conducted in order to characterize newly synthesized steroid derivatives for further investigation regarding their antiproliferative and antimicrobial activity. This article is based upon work from COST Action (CM1306), supported by COST (European Cooperation in Science and Technology).

Keywords: antiproliferative activity, chromatographic lipophilicity, liquid chromatography, steroids

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20 Association of Mir-196a Expression in Esophageal Tissue with Barrett´s Esophagus and Esophageal Adenocarcinoma

Authors: Petra Borilova Linhartova, Michaela Ruckova, Sabina Sevcikova, Natalie Mlcuchova, Jan Bohm, Katerina Zukalova, Monika Vlachova, Jiri Dolina, Lumir Kunovsky, Radek Kroupa, Zdenek Pavlovsky, Zdenek Danek, Tereza Deissova, Lydie Izakovicova Holla, Ondrej Slaby, Zdenek Kala

Abstract:

Esophageal adenocarcinoma (EAC) is a highly aggressive malignancy that frequently develops from Barrett's esophagus (BE), a premalignant pathologic change occurring in the lower end of the esophagus. Specific microRNAs (miRNAs), small non-coding RNAs that function as posttranscriptional regulators of gene expression, were repeatedly proved to play key roles in the pathogenesis of these diseases. This pilot study aimed to analyze four selected miRNAs in esophageal tissues from healthy controls (HC) and patients with reflux esophagitis (RE)/BE/EAC, as well as to compare expression at the site of Barrett's mucosa/adenocarcinoma and healthy esophageal tissue outside the area of the main pathology in patients with BE/EAC. In this pilot study, 22 individuals (3 HC, 8 RE, 5 BE, 6 EAC) were included and endoscopically examined. RNA was isolated from the fresh-frozen esophageal tissue (stored in the RNAlater™ Stabilization Solution −70°C) using the AllPrep DNA/RNA/miRNA Universal Kit. Subsequent RT-qPCR analysis was performed using selected TaqMan MicroRNA Assays for miR-21, miR-34a, miR-196a, miR-196b, and endogenous control (RNU44). While the expression of miR-21 in the esophageal tissue with the main pathology was decreased in BE and EAC patients in comparison to the group of HC and RE patients (p=0.01), the expression of miR-196a was increased in the BE and EAC patients (p<0.01). Correlations between those miRNAs expression in tissue and severity of diagnosis were observed (p<0.05). In addition, miR-196a was significantly more expressed at the site with the main pathology than in paired adjacent esophageal tissue in BE and EAC patients (p<0.01). In conclusion, our pilot results showed that miR-196a, which regulates the proliferation, invasion, and migration (and was previously associated with esophageal squamous cell carcinoma and marked as a potential therapeutic target), could be a diagnostic tissue biomarker for BE and EAC as well.

Keywords: microRNA, barrett´s esophagus, esophageal adenocarcinoma, biomarker

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19 An Ecosystem Approach to Natural Resource Management: Case Study of the Topčiderska River, Serbia

Authors: Katarina Lazarević, Mirjana Todosijević, Tijana Vulević, Natalija Momirović, Ranka Erić

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Due to increasing demand, climate change, and world population growth, natural resources are getting exploit fast. One of the most important natural resources is soil, which is susceptible to degradation. Erosion as one of the forms of land degradation is also one of the most global environmental problems. Ecosystem services are often defined as benefits that nature provides to humankind. Soil, as the foundation of basic ecosystem functions, provides benefits to people, erosion control, water infiltration, food, fuel, fibers… This research is using the ecosystem approach as a strategy for natural resources management for promoting sustainability and conservation. The research was done on the Topčiderska River basin (Belgrade, Serbia). The InVEST Sediment Delivery Ratio model was used, to quantify erosion intensity with a spatial distribution output map of overland sediment generation and delivery to the stream. InVEST SDR, a spatially explicit model, is using a method based on the concept of hydrological connectivity and (R) USLE model. This, combined with socio-economic and law and policy analysis, gives a full set of information to decision-makers helping them to successfully manage and deliver sustainable ecosystems.

Keywords: ecosystem services, InVEST model, soil erosion, sustainability

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18 Exposure to Violence and Its Association with Mental Health in Swedish Prospective Exchange Students and Campus Students

Authors: Emil Danehorn, Ulla Peterson, Marie Oscarsson, Goldina Smirthwaite, Katarina Swahnberg

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Swedish students who choose to study abroad are common, but there has been limited research on exchange students. There are indications that some Swedish exchange students have been exposed to violence, but it is not known if they, already before their semester abroad, are more exposed to violence than students who remain on campus. Our aim was to investigate potential differences in exposure to physical, sexual, and emotional violence between prospective exchange students and campus students and between male and female students in general, as well as to determine any associations between mental health and exposure to violence among students in the total sample. Method: Comparative cross-sectional design using an online survey containing the NorVold Abuse Questionnaire and the General Health Questionnaire 12. A total of 565 participants took part in the study. Results: Prospective exchange students were less exposed to emotional violence compared to campus students. About one-half of all the participating female students had been exposed to sexual violence and about one-third of all the male students had been exposed to physical violence. The results also indicated that exposure to more than one form of violence was associated with poor mental health for students as a group. Conclusion: Prospective exchange students as a group do not report more exposure to physical and sexual violence than campus students but do report less exposure to emotional violence. However, the overall frequency of exposure to violence among the students was high. This highlights the need for proactive efforts for everyone, not least for exchange students who will be staying in a new environment unknown to them.

Keywords: sexual violence, physical violence, emotional violence, exchange students, mental health

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17 Microbiological Activity and Molecular Docking Study of Selected Steroid Derivatives of Biomedical Importance

Authors: Milica Karadzic, Lidija Jevric, Sanja Podunavac-Kuzmanovic, Strahinja Kovacevic, Sinisa Markov, Aleksandar Okljesa, Andrea Nikolic, Marija Sakac, Katarina Penov Gasi

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This study considered the microbiological activity determination and molecular docking study for selected steroid derivatives of biomedical importance. Minimal inhibitory concentration (MIC) was determined for steroid derivatives against Staphylococcus aureus using macrodilution method. Some of the investigated steroid derivatives express bacteriostatic effect against Staphylococcus aureus. Molecular docking approaches are the most widely used techniques for predicting the binding mode of a ligand. Molecular docking study was done for steroid derivatives for androgen receptor negative prostate cancer cell line (PC-3) toward Human Cytochrome P450 CYP17A1. The molecules that had the smallest experimental IC50 values confirmed their ability to dock into active place using suitable molecular docking procedure. The binding disposition of those molecules was thoroughly investigated. Microbiological analysis and molecular docking study were conducted with aim to additionally characterize selected steroid derivatives for future investigation regarding their biological activity and to estimate the binding-affinities of investigated derivatives. This article is based upon work from COST Action (TD1305), supported by COST (European Cooperation and Science and Technology).

Keywords: binding affinity, minimal inhibitory concentration, molecular docking, pc-3 cell line, staphylococcus aureus, steroids

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16 Non-Linear Assessment of Chromatographic Lipophilicity and Model Ranking of Newly Synthesized Steroid Derivatives

Authors: Milica Karadzic, Lidija Jevric, Sanja Podunavac-Kuzmanovic, Strahinja Kovacevic, Anamarija Mandic, Katarina Penov Gasi, Marija Sakac, Aleksandar Okljesa, Andrea Nikolic

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The present paper deals with chromatographic lipophilicity prediction of newly synthesized steroid derivatives. The prediction was achieved using in silico generated molecular descriptors and quantitative structure-retention relationship (QSRR) methodology with the artificial neural networks (ANN) approach. Chromatographic lipophilicity of the investigated compounds was expressed as retention factor value logk. For QSRR modeling, a feedforward back-propagation ANN with gradient descent learning algorithm was applied. Using the novel sum of ranking differences (SRD) method generated ANN models were ranked. The aim was to distinguish the most consistent QSRR model that can be found, and similarity or dissimilarity between the models that could be noticed. In this study, SRD was performed with average values of retention factor value logk as reference values. An excellent correlation between experimentally observed retention factor value logk and values predicted by the ANN was obtained with a correlation coefficient higher than 0.9890. Statistical results show that the established ANN models can be applied for required purpose. This article is based upon work from COST Action (TD1305), supported by COST (European Cooperation in Science and Technology).

Keywords: artificial neural networks, liquid chromatography, molecular descriptors, steroids, sum of ranking differences

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15 Non-Linear Assessment of Chromatographic Lipophilicity of Selected Steroid Derivatives

Authors: Milica Karadžić, Lidija Jevrić, Sanja Podunavac-Kuzmanović, Strahinja Kovačević, Anamarija Mandić, Aleksandar Oklješa, Andrea Nikolić, Marija Sakač, Katarina Penov Gaši

Abstract:

Using chemometric approach, the relationships between the chromatographic lipophilicity and in silico molecular descriptors for twenty-nine selected steroid derivatives were studied. The chromatographic lipophilicity was predicted using artificial neural networks (ANNs) method. The most important in silico molecular descriptors were selected applying stepwise selection (SS) paired with partial least squares (PLS) method. Molecular descriptors with satisfactory variable importance in projection (VIP) values were selected for ANN modeling. The usefulness of generated models was confirmed by detailed statistical validation. High agreement between experimental and predicted values indicated that obtained models have good quality and high predictive ability. Global sensitivity analysis (GSA) confirmed the importance of each molecular descriptor used as an input variable. High-quality networks indicate a strong non-linear relationship between chromatographic lipophilicity and used in silico molecular descriptors. Applying selected molecular descriptors and generated ANNs the good prediction of chromatographic lipophilicity of the studied steroid derivatives can be obtained. This article is based upon work from COST Actions (CM1306 and CA15222), supported by COST (European Cooperation and Science and Technology).

Keywords: artificial neural networks, chemometrics, global sensitivity analysis, liquid chromatography, steroids

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14 Evaluation of Newly Synthesized Steroid Derivatives Using In silico Molecular Descriptors and Chemometric Techniques

Authors: Milica Ž. Karadžić, Lidija R. Jevrić, Sanja Podunavac-Kuzmanović, Strahinja Z. Kovačević, Anamarija I. Mandić, Katarina Penov-Gaši, Andrea R. Nikolić, Aleksandar M. Oklješa

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This study considered selection of the in silico molecular descriptors and the models for newly synthesized steroid derivatives description and their characterization using chemometric techniques. Multiple linear regression (MLR) models were established and gave the best molecular descriptors for quantitative structure-retention relationship (QSRR) modeling of the retention of the investigated molecules. MLR models were without multicollinearity among the selected molecular descriptors according to the variance inflation factor (VIF) values. Used molecular descriptors were ranked using generalized pair correlation method (GPCM). In this method, the significant difference between independent variables can be noticed regardless almost equal correlation between dependent variable. Generated MLR models were statistically and cross-validated and the best models were kept. Models were ranked using sum of ranking differences (SRD) method. According to this method, the most consistent QSRR model can be found and similarity or dissimilarity between the models could be noticed. In this study, SRD was performed using average values of experimentally observed data as a golden standard. Chemometric analysis was conducted in order to characterize newly synthesized steroid derivatives for further investigation regarding their potential biological activity and further synthesis. This article is based upon work from COST Action (CM1105), supported by COST (European Cooperation in Science and Technology).

Keywords: generalized pair correlation method, molecular descriptors, regression analysis, steroids, sum of ranking differences

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13 The Effectiveness of Kinesiotaping Methods in Rehabilitation Therapy

Authors: Ana-Katarina Nikich

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Background: The kinesiotaping method is often used in physiotherapy and rehabilitation. The purpose of this study was to evaluate the effectiveness of taping in the rehabilitation process of patients. Materials and methods: The study involved 90 male and female patients (the average age was 40-50 years) with various conditions requiring rehabilitation, such as injuries of the musculoskeletal system, sports injuries and other ailments. All patients were divided into two groups: experimental (n=40) and control (n=50). Both groups received 20 days of standard rehabilitation. In the experimental group, kinesiotaping methods were used, taking into account the individual characteristics of each patient. The control group performed regular exercises and physical therapy, but without using kinesiotape. During the study, physical parameters were monitored, interviews were conducted and the conditions of patients from both groups were compared. Results and discussion: The use of the kinesiotaping method in the rehabilitation process led to a significant improvement in physical parameters and pain reduction in patients. Significant improvement (p <0.005) was observed in all evaluated parameters among the patients of the experimental group. The control group also showed sufficient improvement (p <0.005), but the percentage of the experimental group was higher. As a result of the observation, the patients of the experimental group showed faster and more complete rehabilitation compared to the control group. The use of the kinesiotaping method allows to reduce the load on the damaged areas, improve blood circulation and lymphatic drainage, as well as increase stability and coordination of movements. Conclusions: Kinesiotaping as one of the modern therapeutic methods has shown its effectiveness in the rehabilitation process, contributing to the optimal recovery of patients with various conditions requiring rehabilitation. The use of tapes should be included in a comprehensive rehabilitation program to achieve the best results and reduce recovery time.

Keywords: kinesiotaping, rehabilitation, therapy, pain

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12 Gray’s Anatomy for Students: First South Asia Edition Highlights

Authors: Raveendranath Veeramani, Sunil Jonathan Holla, Parkash Chand, Sunil Chumber

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Gray’s Anatomy for Students has been a well-appreciated book among undergraduate students of anatomy in Asia. However, the current curricular requirements of anatomy require a more focused and organized approach. The editors of the first South Asia edition of Gray’s Anatomy for Students hereby highlight the modifications and importance of this edition. There is an emphasis on active learning by making the clinical relevance of anatomy explicit. Learning anatomy in context has been fostered by the association between anatomists and clinicians in keeping with the emerging integrated curriculum of the 21st century. The language has been simplified to aid students who have studied in the vernacular. The original illustrations have been retained, and few illustrations have been added. There are more figure numbers mentioned in the text to encourage students to refer to the illustrations while learning. The text has been made more student-friendly by adding generalizations, classifications and summaries. There are useful review materials at the beginning of the chapters which include digital resources for self-study. There are updates on imaging techniques to encourage students to appreciate the importance of essential knowledge of the relevant anatomy to interpret images, due emphasis has been laid on dissection. Additional importance has been given to the cranial nerves, by describing their relevant details with several additional illustrations and flowcharts. This new edition includes innovative features such as set inductions, outlines for subchapters and flowcharts to facilitate learning. Set inductions are mostly clinical scenarios to create interest in the need to study anatomy for healthcare professions. The outlines are a modern multimodal facilitating approach towards various topics to empower students to explore content and direct their learning and include learning objectives and material for review. The components of the outline encourage the student to be aware of the need to create solutions to clinical problems. The outlines help students direct their learning to recall facts, demonstrate and analyze relationships, use reason to explain concepts, appreciate the significance of structures and their relationships and apply anatomical knowledge. The 'structures to be identified in a dissection' are given as Level I, II and III which represent the 'must know, desirable to know and nice to know' content respectively. The flowcharts have been added to get an overview of the course of a structure, recapitulate important details about structures, and as an aid to recall. There has been a great effort to balance the need to have content that would enable students to understand concepts as well as get the basic material for the current condensed curriculum.

Keywords: Grays anatomy, South Asia, human anatomy, students anatomy

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11 Cytolethal Distending Toxins in Intestinal and Extraintestinal E. coli

Authors: Katarína Čurová, Leonard Siegfried, Radka Vargová, Marta Kmeťová, Vladimír Hrabovský

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Introduction: Cytolethal distending toxins (CDTs) represent intracellular acting proteins which interfere with cell cycle of eukaryotic cells. They are produced by Gram-negative bacteria with afinity to mucocutaneous surfaces and could play a role in the pathogenesis of various diseases. CDTs induce DNA damage probably through DNAse activity, which causes cell cycle arrest and leads to further changes (cell distension and death, apoptosis) depending on the cell type. Five subtypes of CDT (I to V) were reported in E. coli. Methods: We examined 252 E. coli strains belonging to four different groups. Of these strains, 57 were isolated from patients with diarrhea, 65 from patients with urinary tract infections (UTI), 65 from patients with sepsis and 65 from patients with other extraintestinal infections (mostly surgical wounds, decubitus ulcers and respiratory tract infections). Identification of these strains was performed by MALDI-TOF analysis and detection of genes encoding CDTs and determination of the phylogenetic group was performed by PCR. Results: In this study, we detected presence of cdt genes in 11 of 252 E. coli strains tested (4,4 %). Four cdt positive E. coli strains were confirmed in group of UTI (6,15 %), three cdt positive E. coli strains in groups of diarrhea (5,3 %) and other extraintestinal infections (4,6 %). The lowest incidence, one cdt positive E. coli strain, was observed in group of sepsis (1,5 %). All cdt positive E. coli strains belonged to phylogenetic group B2. Conclusion: CDT-producing E. coli are isolated in a low percentage from patients with intestinal and extraintestinal infections, including sepsis and our results correspond with these studies. A weak prevalence of cdt genes suggests that CDTs are not major virulence factors but in combination with other virulence factors may increase virulence potential of E. coli. We suppose that all 11 cdt positive E. coli strains represent real pathogens because they belong to the phylogenetic group B2 which is pathogenic lineage for bacteria E. coli.

Keywords: cytolethal distending toxin, E. coli, phylogenetic group, extraintestinal infection, diarrhea

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10 A Photoredox (C)sp³-(C)sp² Coupling Method Comparison Study

Authors: Shasline Gedeon, Tiffany W. Ardley, Ying Wang, Nathan J. Gesmundo, Katarina A. Sarris, Ana L. Aguirre

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Drug discovery and delivery involve drug targeting, an approach that helps find a drug against a chosen target through high throughput screening and other methods by way of identifying the physical properties of the potential lead compound. Physical properties of potential drug candidates have been an imperative focus since the unveiling of Lipinski's Rule of 5 for oral drugs. Throughout a compound's journey from discovery, clinical phase trials, then becoming a classified drug on the market, the desirable properties are optimized while minimizing/eliminating toxicity and undesirable properties. In the pharmaceutical industry, the ability to generate molecules in parallel with maximum efficiency is a substantial factor achieved through sp²-sp² carbon coupling reactions, e.g., Suzuki Coupling reactions. These reaction types allow for the increase of aromatic fragments onto a compound. More recent literature has found benefits to decreasing aromaticity, calling for more sp³-sp² carbon coupling reactions instead. The objective of this project is to provide a comparison between various sp³-sp² carbon coupling methods and reaction conditions, collecting data on production of the desired product. There were four different coupling methods being tested amongst three cores and 4-5 installation groups per method; each method ran under three distinct reaction conditions. The tested methods include the Photoredox Decarboxylative Coupling, the Photoredox Potassium Alkyl Trifluoroborate (BF3K) Coupling, the Photoredox Cross-Electrophile (PCE) Coupling, and the Weix Cross-Electrophile (WCE) Coupling. The results concluded that the Decarboxylative method was very difficult in yielding product despite the several literature conditions chosen. The BF3K and PCE methods produced competitive results. Amongst the two Cross-Electrophile coupling methods, the Photoredox method surpassed the Weix method on numerous accounts. The results will be used to build future libraries.

Keywords: drug discovery, high throughput chemistry, photoredox chemistry, sp³-sp² carbon coupling methods

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9 Fuzzy Decision Making to the Construction Project Management: Glass Facade Selection

Authors: Katarina Rogulj, Ivana Racetin, Jelena Kilic

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In this study, the fuzzy logic approach (FLA) was developed for construction project management (CPM) under uncertainty and duality. The focus was on decision making in selecting the type of the glass facade for a residential-commercial building in the main design. The adoption of fuzzy sets was capable of reflecting construction managers’ reliability level over subjective judgments, and thus the robustness of the system can be achieved. An α-cuts method was utilized for discretizing the fuzzy sets in FLA. This method can communicate all uncertain information in the optimization process, taking into account the values of this information. Furthermore, FLA provides in-depth analyses of diverse policy scenarios that are related to various levels of economic aspects when it comes to the construction projects' valid decision making. The developed approach is applied to CPM to demonstrate its applicability. Analyzing the materials of glass facades, variants were defined. The development of the FLA for the CPM included relevant construction projec'ts stakeholders that were involved in the criteria definition to evaluate each variant. Using fuzzy Decision-Making Trial and Evaluation Laboratory Method (DEMATEL) comparison of the glass facade was conducted. This way, a rank, according to the priorities for inclusion into the main design, of variants is obtained. The concept was tested on a residential-commercial building in the city of Rijeka, Croatia. The newly developed methodology was then compared with the existing one. The aim of the research was to define an approach that will improve current judgments and decisions when it comes to the material selection of buildings facade as one of the most important architectural and engineering tasks in the main design. The advantage of the new methodology compared to the old one is that it includes the subjective side of the managers’ decisions, as an inevitable factor in each decision making. The proposed approach can help construction projects managers to identify the desired type of glass facade according to their preference and practical conditions, as well as facilitate in-depth analyses of tradeoffs between economic efficiency and architectural design.

Keywords: construction projects management, DEMATEL, fuzzy logic approach, glass façade selection

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8 Automatic Identification of Aquatic Insects Based on Deep Learning and Computer Vision

Authors: Predrag Simović, Katarina Stojanović, Milena Radenković, Dimitrija Savić Zdravković, Aleksandar Milosavljević, Bratislav Predić, Milenka Božanić, Ana Petrović, Djuradj Milošević

Abstract:

Mayflies (Ephemeroptera), stoneflies (Plecoptera), and caddisflies (Trichoptera) (collectively referred to as EPT) are key participants in most freshwater habitats and often exhibit high diversity. Moreover, their presence and relative abundance are used in freshwater ecological and biomonitoring studies. Current methods for freshwater ecosystem biomonitoring follow a traditional approach of taxa monitoring based on morphological characters, which is time-consuming, and often generates data sets with low taxonomic resolution and unverifiable identification precision. To assist in solving identification problems and contribute to the knowledge of the distribution of many species, there was a need to develop alternative approaches in macroinvertebrate sample identification. Here, we establish an automatic machine-based identification approach for EPT taxa (Insect) using deep Convolutional Neural Networks (CNNs) and computer vision to increase the efficiency and taxonomic resolution in biomonitoring. The 5 550 specimens were collected from freshwater ecosystems of Serbia, and the deep model was built upon 90 EPT taxa. The protocol for obtaining images included the following stages: taxonomic identification by human experts and DNA barcoding validation, mounting the larvae, and photographing the dorsal side using a stereomicroscope and camera (16 650 individuals). The most informative image regions (the dorsal segments of individuals) for the decision-making process in the deep learning model were visualized using Gradient Weighted Class Activation Mapping (Grad-CAM). After training the artificial neural network, a CNN model was then built that was able to classify the 90 EPT taxa into their respective taxonomic categories automatically with 98.7%. Our approach offers a straightforward and efficient solution for routine monitoring programs, focusing on key biotic descriptors, such as EPT taxa. In addition, this application provides a streamlined solution that not only saves time, reduces equipment and expert requirements but also significantly enhances reliability and information content. The identification of the EPT larvae is difficult because of the variation of morphological features even within a single genus or the close resemblance of several species, and therefore, future research should focus on increasing the number of entities (species) in the model.

Keywords: convolutional neural networks, DNA barcoding, EPT taxa, biomonitoring

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7 Energy Atlas: Geographic Information Systems-Based Energy Analysis and Planning Tool

Authors: Katarina Pogacnik, Ursa Zakrajsek, Nejc Sirk, Ziga Lampret

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Due to an increase in living standards along with global population growth and a trend of urbanization, municipalities and regions are faced with an ever rising energy demand. A challenge has arisen for cities around the world to modify the energy supply chain in order to reduce its consumption and CO₂ emissions. The aim of our work is the development of a computational-analytical platform for dynamic support in decision-making and the determination of economic and technical indicators of energy efficiency in a smart city, named Energy Atlas. Similar products in this field focuse on a narrower approach, whereas in order to achieve its aim, this platform encompasses a wider spectrum of beneficial and important information for energy planning on a local or regional scale. GIS based interactive maps provide an extensive database on the potential, use and supply of energy and renewable energy sources along with climate, transport and spatial data of the selected municipality. Beneficiaries of Energy atlas are local communities, companies, investors, contractors as well as residents. The Energy Atlas platform consists of three modules named E-Planning, E-Indicators and E-Cooperation. The E-Planning module is a comprehensive data service, which represents a support towards optimal decision-making and offers a sum of solutions and feasibility of measures and their effects in the area of efficient use of energy and renewable energy sources. The E-Indicators module identifies, collects and develops optimal data and key performance indicators and develops an analytical application service for dynamic support in managing a smart city in regards to energy use and sustainable environment. In order to support cooperation and direct involvement of citizens of the smart city, the E-cooperation is developed with the purpose of integrating the interdisciplinary and sociological aspects of energy end-users. Interaction of all the above-described modules contributes to regional development because it enables for a precise assessment of the current situation, strategic planning, detection of potential future difficulties and also the possibility of public involvement in decision-making. From the implementation of the technology in Slovenian municipalities of Ljubljana, Piran, and Novo mesto, there is evidence to suggest that the set goals are to be achieved to a great extent. Such thorough urban energy planning tool is viewed as an important piece of the puzzle towards achieving a low-carbon society, circular economy and therefore, sustainable society.

Keywords: circular economy, energy atlas, energy management, energy planning, low-carbon society

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6 Effect of Copper Complexes on Human Colon Carcinoma Cell Line and Human Breast Carcinoma Cell Line

Authors: Katarína Koňariková, Georgios A. Perdikaris, Lucia Andrezálová, Zdeňka Ďuračková, Lucia Laubertová, Helena Gbelcová, Ingrid Žitňanová

Abstract:

Introduction: The continuous demand for new anti-cancer drugs has stimulated chemotherapeutic research based on the use of essential metalloelements with the aim to develop potential drugs with lower toxicity and higher antiproliferative activity against tumors. Copper(II) and its complexes play an important role as suitable species for antiproliferative tests. Objectives: The central objective of the current study was to investigate the potential in vitro anti-proliferative effects of N-salicylidene-L-glutamato copper (II) complexes and molecular mechanism of apoptosis induced by tested complexes. In our project we tested N-salicylidene-L-glutamato copper (II) complexes ZK1 - [Cu(N-salicylidene-L-glutamato)(H2O)2].H2O; MK0 - ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O); MK1 - [Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O; MK3 - transbis(ethanol)tetrakis(imidazol)Cu(II)(2+)bis(N-salicylidene-D,L-glutamato-N,O)-KO:KO´-(imidazol); MK5 - [Cu(N-salicylidene-D,L- glutamato)(2-methylimidazol] at concentration range 0.001-100 µmol/L against human colon carcinoma cell line HT-29 and human breast carcinoma cell line MCF-7. Methods: Viability was assessed by direct counting of 0.4% trypan blue dye-excluding cells after 24, 48 and 72 hour cultivations with or without copper complex and by MTT assay. To analyze the type of cell death and its mechanism induced by our copper complex we used different methods. To distinguish apoptosis from necrosis we used electrophoretic analysis, to study the activity of caspases 8 and 9 – luminometric analysis and caspase activity 3 colorimetric assay. Results: The observed anti-proliferative effect of the copper complexes appeared to be dose-, time- and cell line- dependent. Human colon carcinoma cells HT-29 appeared to be more sensitive to the complex MK0 ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O) than to ZK1 ([Cu(N-salicylidene-L-glutamato)(H2O)2].H2O) and MK1 ([Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O)). Human colon carcinoma cells HT-29 appeared to be more sensitive to the complex than human breast carcinoma cells MCF-7. IC50 decreased with time of incubation (24, 48 and 72h) for HT-29, but increased for MCF-7. By electrophoresis we found apoptotic cell death induced by our copper complexes in HT-29 at concentrations 1, 10, 50 and 100 µmol/L after 48h (ZK1) and 72h (MK0, MK1) and in MCF-7 we did not find apoptosis. We also studied molecular mechanism of apoptosis in HT-29 induced by copper complexes. We found active caspase 9 in HT-29 after ZK1 ([Cu(N-salicylidene-L-glutamato)(H2O)2].H2O) and MK1 ([Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O)) influence and active caspase 8 after MK0 ([Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O) influence. Conclusion: Our copper complexes showed cytotoxic activities against human colon carcinoma cells HT-29 and breast cancer cell line MCF-7 in vitro. Apoptosis was activated by mitochondrial pathway (intrinsic pathway) in case of ZK1 [Cu(N-salicylidene-L-glutamato)(H2O)2].H2O; MK1 [Cu(N-salicylidene-5-methyl-L-glutamato)(H2O)].H2O; MK3 - transbis(ethanol)tetrakis(imidazol)Cu(II)(2+)bis(N-salicylidene-D,L-glutamato-N,O)-KO:KO´-(imidazol) and MK5 - [Cu(N-salicylidene-D,L- glutamato)(2-methylimidazol] copper complexes and by death receptors (extrinsic pathway) in case of MK0 [Cu2(N-sal-D,L-glu)2(isoquinoline)2].2H2O copper complex in HT-29.

Keywords: apoptosis, copper complex, cancer, carcinoma cell line

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5 Technology in Commercial Law Enforcement: Tanzania, Canada, and Singapore Comparatively

Authors: Katarina Revocati Mteule

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The background of this research arises from global demands for fair business opportunities. As one of responses to these demands, nations embarked on reforms in commercial laws. In 1990s Tanzania resorted to economic transformation through liberalization to attract more investments included reform in commercial laws enforcement. This research scrutinizes the effectiveness of reforms in Tanzania in comparison with Canada and Singapore and the role of technology. The methodology to be used is doctrinal legal research mixed with international comparative legal research. It involves comparative analysis of library, online, and internet resources as well as Case Laws and Statutory Laws. Tanzania, Canada and Singapore are sampled comparators basing on their distinct level of economic development. The criteria of analysis includes the nature of reforms, type of technology, technological infrastructure and human resource technical competence in each country. As the world progresses towards reforms in commercial laws, improvements in law, policy, and regulatory frameworks are paramount. Specifically, commercial laws are essential in contract enforcement and dispute resolution and how it copes with modern technologies is a concern. Harnessing the best technology is necessary to cope with the modernity in world businesses. In line with this, Tanzania is improving its business environment, including law enforcement mechanisms that are supportive to investments. Reforms such as specialized commercial law enforcement coupled with alternative dispute resolutions such as arbitration, mediation, and reconciliation are emphasized. Court technology as one of the reform tools given high priority. This research evaluates the progress and the effectiveness of the reforms in Commercial Laws towards friendly business environment in Tanzania in comparison with Canada and Singapore. The experience of Tanzania is compared with Canada and Singapore to see what to improve for each country to enhance quick and fair enforcement of commercial law. The research proposes necessary global standards of procedures and in national laws to offer a business-friendly environment and the use of appropriate technology. Solutions are proposed in tackling the challenges of delays in enforcing Commercial Laws such as case management, funding, legal and procedural hindrances, laxity among staff, and abuse of Court process among litigants, all in line with modern technology. It is the finding of the research that proper use of technology has managed to reduce case backlogs and time taken to resolve a commercial dispute, to increase court integrity by minimizing human contacts in commercial law enforcement which may lead to solicitation of favors and saving of parties’ time due to online service. Among the three countries, each one is facing a distinct challenge due to the level of poverty and remoteness from online service. How solutions are found in one country is a lesson to another. To conclude, this paper is suggesting solutions for improving the commercial law enforcement mechanisms in line with modern technology. The call for technological transformation is essential for the enforcement of commercial laws.

Keywords: commercial law, enforcement, technology

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4 Residents' Incomes in Local Government Unit as the Major Determinant of Local Budget Transparency in Croatia: Panel Data Analysis

Authors: Katarina Ott, Velibor Mačkić, Mihaela Bronić, Branko Stanić

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The determinants of national budget transparency have been widely discussed in the literature, while research on determinants of local budget transparency are scarce and empirically inconclusive, particularly in the new, fiscally centralised, EU member states. To fill the gap, we combine two strands of the literature: that concerned with public administration and public finance, shedding light on the economic and financial determinants of local budget transparency, and that on the political economy of transparency (principal agent theory), covering the relationships among politicians and between politicians and voters. Our main hypothesis states that variables describing residents’ capacity have a greater impact on local budget transparency than variables indicating the institutional capacity of local government units (LGUs). Additional subhypotheses test the impact of each variable analysed on local budget transparency. We address the determinants of local budget transparency in Croatia, measured by the number of key local budget documents published on the LGUs’ websites. By using a data set of 128 cities and 428 municipalities over the 2015-2017 period and by applying panel data analysis based on Poisson and negative binomial distribution, we test our main hypothesis and sub-hypotheses empirically. We measure different characteristics of institutional and residents’ capacity for each LGU. Age, education and ideology of the mayor/municipality head, political competition indicators, number of employees, current budget revenues and direct debt per capita have been used as a measure of the institutional capacity of LGU. Residents’ capacity in each LGU has been measured through the numbers of citizens and their average age as well as by average income per capita. The most important determinant of local budget transparency is average residents' income per capita at both city and municipality level. The results are in line with most previous research results in fiscally decentralised countries. In the context of a fiscally centralised country with numerous small LGUs, most of whom have low administrative and fiscal capacity, this has a theoretical rationale in the legitimacy and principal-agent theory (opportunistic motives of the incumbent). The result is robust and significant, but because of the various other results that change between city and municipality levels (e.g. ideology and political competition), there is a need for further research (both on identifying other determinates and/or methods of analysis). Since in Croatia the fiscal capacity of a LGU depends heavily on the income of its residents, units with higher per capita incomes in many cases have also higher budget revenues allowing them to engage more employees and resources. In addition, residents’ incomes might be also positively associated with local budget transparency because of higher citizen demand for such transparency. Residents with higher incomes expect more public services and have more access to and experience in using the Internet, and will thus typically demand more budget information on the LGUs’ websites.

Keywords: budget transparency, count data, Croatia, local government, political economy

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3 Evaluation of Herbal Extracts for Their Potential Application as Skin Prebiotics

Authors: Anja I. Petrov, Milica B. Veljković, Marija M. Ćorović, Ana D. Milivojević, Milica B. Simović, Katarina M. Banjanac, Dejan I. Bezbradica

Abstract:

One of the fundamental requirements for overall human well-being is a stable and balanced microbiome. Aside from the microorganisms that reside within the body, a large number of microorganisms, especially bacteria, swarming the human skin is in homeostasis with the host and represents a skin microbiota. Even though the immune system of the skin is capable of distinguishing between commensal and potentially harmful transient bacteria, the cutaneous microbial balance can be disrupted under certain circumstances. In that case, a reduction in the skin microbiota diversity, as well as changes in metabolic activity, results in dermal infections and inflammation. Probiotics and prebiotics have the potential to play a significant role in the treatment of these skin disorders. The most common resident bacteria found on the skin, Staphylococcus epidermidis, can act as a potential skin probiotic, contributing to the protection of healthy skin from pathogen colonization, such as Staphylococcus aureus, which is related to atopic dermatitis exacerbation. However, as it is difficult to meet regulations in cosmetic products, another therapy approach could be topical prebiotic supplementation of the skin microbiota. In recent research, polyphenols are attracting scientists' interest as biomolecules with possible prebiotic effects on the skin microbiota. This research aimed to determine how herbal extracts rich in different polyphenolic compounds (lemon balm, St. John's wort, coltsfoot, pine needle, and yarrow) affected the growth of S. epidermidis and S. aureus. The first part of the study involved screening plants to determine if they could be regarded as probable candidates to be skin prebiotics. The effect of each plant on bacterial growth was examined by supplementing the nutrient medium with their extracts and comparing it with control samples (without extract). The results obtained after 24 h of incubation showed that all tested extracts influenced the growth of the examined bacteria to some extent. Since lemon balm and St. John's wort extracts displayed bactericidal activity against S. epidermidis, whereas coltsfoot inhibited both bacteria equally, they were not explored further. On the other hand, pine needles and yarrow extract led to an increase in S. epidermidis/S. aureus ratio, making them prospective candidates to be used as skin prebiotics. By examining the prebiotic effect of two extracts at different concentrations, it was revealed that, in the case of yarrow, 0.1% of extract dry matter in the fermentation medium was optimal, while for the pine needle extract, a concentration of 0.05% was preferred, since it selectively stimulated S. epidermidis growth and inhibited S. aureus proliferation. Additionally, the total polyphenols and flavonoid content of the two extracts were determined, revealing different concentrations and polyphenol profiles. Since yarrow and pine extracts affected the growth of skin bacteria in a dose-dependent manner, by carefully selecting the quantities of these extracts, and thus polyphenols content, it is possible to achieve desirable alterations of skin microbiota composition, which may be suitable for the treatment of atopic dermatitis.

Keywords: herbal extracts, polyphenols, skin microbiota, skin prebiotics

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2 Sorbitol Galactoside Synthesis Using β-Galactosidase Immobilized on Functionalized Silica Nanoparticles

Authors: Milica Carević, Katarina Banjanac, Marija ĆOrović, Ana Milivojević, Nevena Prlainović, Aleksandar Marinković, Dejan Bezbradica

Abstract:

Nowadays, considering the growing awareness of functional food beneficial effects on human health, due attention is dedicated to the research in the field of obtaining new prominent products exhibiting improved physiological and physicochemical characteristics. Therefore, different approaches to valuable bioactive compounds synthesis have been proposed. β-Galactosidase, for example, although mainly utilized as hydrolytic enzyme, proved to be a promising tool for these purposes. Namely, under the particular conditions, such as high lactose concentration, elevated temperatures and low water activities, reaction of galactose moiety transfer to free hydroxyl group of the alternative acceptor (e.g. different sugars, alcohols or aromatic compounds) can generate a wide range of potentially interesting products. Up to now, galacto-oligosaccharides and lactulose have attracted the most attention due to their inherent prebiotic properties. The goal of this study was to obtain a novel product sorbitol galactoside, using the similar reaction mechanism, namely transgalactosylation reaction catalyzed by β-galactosidase from Aspergillus oryzae. By using sugar alcohol (sorbitol) as alternative acceptor, a diverse mixture of potential prebiotics is produced, enabling its more favorable functional features. Nevertheless, an introduction of alternative acceptor into the reaction mixture contributed to the complexity of reaction scheme, since several potential reaction pathways were introduced. Therefore, the thorough optimization using response surface method (RSM), in order to get an insight into different parameter (lactose concentration, sorbitol to lactose molar ratio, enzyme concentration, NaCl concentration and reaction time) influences, as well as their mutual interactions on product yield and productivity, was performed. In view of product yield maximization, the obtained model predicted optimal lactose concentration 500 mM, the molar ratio of sobitol to lactose 9, enzyme concentration 0.76 mg/ml, concentration of NaCl 0.8M, and the reaction time 7h. From the aspect of productivity, the optimum substrate molar ratio was found to be 1, while the values for other factors coincide. In order to additionally, improve enzyme efficiency and enable its reuse and potential continual application, immobilization of β-galactosidase onto tailored silica nanoparticles was performed. These non-porous fumed silica nanoparticles (FNS)were chosen on the basis of their biocompatibility and non-toxicity, as well as their advantageous mechanical and hydrodinamical properties. However, in order to achieve better compatibility between enzymes and the carrier, modifications of the silica surface using amino functional organosilane (3-aminopropyltrimethoxysilane, APTMS) were made. Obtained support with amino functional groups (AFNS) enabled high enzyme loadings and, more importantly, extremely high expressed activities, approximately 230 mg proteins/g and 2100 IU/g, respectively. Moreover, this immobilized preparation showed high affinity towards sorbitol galactoside synthesis. Therefore, the findings of this study could provided a valuable contribution to the efficient production of physiologically active galactosides in immobilized enzyme reactors.

Keywords: β-galactosidase, immobilization, silica nanoparticles, transgalactosylation

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1 Selective Immobilization of Fructosyltransferase onto Glutaraldehyde Modified Support and Its Application in the Production of Fructo-Oligosaccharides

Authors: Milica B. Veljković, Milica B. Simović, Marija M. Ćorović, Ana D. Milivojević, Anja I. Petrov, Katarina M. Banjanac, Dejan I. Bezbradica

Abstract:

In recent decades, the scientific community has recognized the growing importance of prebiotics, and therefore, numerous studies are focused on their economic production due to their low presence in natural resources. It has been confirmed that prebiotics is a source of energy for probiotics in the gastrointestinal tract (GIT) and enable their proliferation, consequently leading to the normal functioning of the intestinal microbiota. Also, products of their fermentation are short-chain fatty acids (SCFA), which play a key role in maintaining and improving the health not only of the GIT but also of the whole organism. Among several confirmed prebiotics, fructooligosaccharides (FOS) are considered interesting candidates for use in a wide range of products in the food industry. They are characterized as low-calorie and non-cariogenic substances that represent an adequate sugar substitute and can be considered suitable for use in products intended for diabetics. The subject of this research will be the production of FOS by transforming sucrose using a fructosyltransferase (FTase) present in commercial preparation Pectinex® Ultra SP-L, with special emphasis on the development of adequate FTase immobilization method that would enable selective isolation of the enzyme responsible for the synthesis of FOS from the complex enzymatic mixture. This would lead to considerable enzyme purification and allow its direct incorporation into different sucrose-based products without the fear that the action of the other hydrolytic enzymes may adversely affect the products' functional characteristics. Accordingly, the possibility of selective immobilization of the enzyme using support with primary amino groups, Purolite® A109, which was previously activated and modified using glutaraldehyde (GA), was investigated. In the initial phase of the research, the effects of individual immobilization parameters such as pH, enzyme concentration, and immobilization time were investigated to optimize the process using support chemically activated with 15% and 0.5% GA to form dimers and monomers, respectively. It was determined that highly active immobilized preparations (371.8 IU/g of support - dimer and 213.8 IU/g of support – monomer) were achieved under acidic conditions (pH 4) provided that an enzyme concentration was 50 mg/g of support after 7 h and 3 h, respectively. Bearing in mind the obtained results of the expressed activity, it is noticeable that the formation of dimers showed higher reactivity compared to the form of monomers. Also, in the case of support modification using 15% GA, the value of the ratio of FTase and pectinase (as dominant enzyme mixture component) activity immobilization yields was 16.45, indicating the high feasibility of selective immobilization of FTase on modified polystyrene resin. After obtaining immobilized preparations of satisfactory features, they were tested in a reaction of FOS synthesis under determined optimal conditions. The maximum FOS yields of approximately 50% of total carbohydrates in the reaction mixture were recorded after 21 h. Finally, it can be concluded that the examined immobilization method yielded highly active, stable and, more importantly, refined enzyme preparation that can be further utilized on a larger scale for the development of continual processes for FOS synthesis, as well as for modification of different sucrose-based mediums.

Keywords: chemical modification, fructooligosaccharides, glutaraldehyde, immobilization of fructosyltransferase

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