Search results for: Alena Kozlová

Authors: Alena Kulikova, Leonid Parmaksiz, Ekaterina Orel

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Social and emotional skills are considered by modern researchers as predictors of a person's success both in specific areas of activity and in the life of a person as a whole. The popularity of this scientific direction ensures the emergence of a large number of practices aimed at developing and evaluating socio-emotional skills. Assessment of social and emotional development is carried out at the national level, as well as at the level of individual regions and institutions. Despite the fact that many of the already existing social and emotional skills assessment tools are quite convenient and reliable, there are now more and more new technologies and task formats which improve the basic characteristics of the tools. Thus, the goal of the current study is to develop a tool for assessing social and emotional skills such as emotion recognition, emotion regulation, empathy and a culture of self-care. To develop a tool assessing social and emotional skills, Rasch-Gutman scenario-based approach was used. This approach has shown its reliability and merit for measuring various complex constructs: parental involvement; teacher practices that support cultural diversity and equity; willingness to participate in the life of the community after psychiatric rehabilitation; educational motivation and others. To assess emotion recognition, we used a situational judgment task based on OCC (Ortony, Clore, and Collins) emotions theory. The main advantage of these two approaches compare to classical Likert scales is that it reduces social desirability in answers. A field test to check the psychometric properties of the developed instrument was conducted. The instrument was developed for the presidential autonomous non-profit organization “Russia - Land of Opportunity” for nationwide soft skills assessment among higher education students. The sample for the field test consisted of 500 people, students aged from 18 to 25 (mean = 20; standard deviation 1.8), 71% female. 67% of students are only studying and are not currently working and 500 employed adults aged from 26 to 65 (mean = 42.5; SD 9), 57% female. Analysis of the psychometric characteristics of the scales was carried out using the methods of IRT (Item Response Theory). A one-parameter rating scale model RSM (Rating scale model) and Graded Response model (GRM) of the modern testing theory were applied. GRM is a polyatomic extension of the dichotomous two-parameter model of modern testing theory (2PL) based on the cumulative logit function for modeling the probability of a correct answer. The validity of the developed scales was assessed using correlation analysis and MTMM (multitrait-multimethod matrix). The developed instrument showed good psychometric quality and can be used by HR specialists or educational management. The detailed results of a psychometric study of the quality of the instrument, including the functioning of the tasks of each scale, will be presented. Also, the results of the validity study by MTMM analysis will be discussed.

Keywords: social and emotional skills, psychometrics, MTMM, IRT

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Authors: Alena Semeradtova, Marcel Stofik, Lucie Mareckova, Petr Maly, Ondrej Stanek, Jan Maly

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Recently, novel strategies based on so-called molecular evolution were shown to be effective for the production of various peptide ligand libraries with high affinities to molecular targets of interest comparable or even better than monoclonal antibodies. The major advantage of these peptide scaffolds is mainly their prevailing low molecular weight and simple structure. This study describes a new high-affinity binding molecules based immunesensor using a simple optical system for human serum albumin (HSA) detection as a model molecule. We present a comparison of two variants of recombinant binders based on albumin binding domain of the protein G (ABD) performed on micropatterned glass chip. Binding domains may be tailored to any specific target of interest by molecular evolution. Micropatterened glass chips were prepared using UV-photolithography on chromium sputtered glasses. Glass surface was modified by (3-aminopropyl)trietoxysilane and biotin-PEG-acid using EDC/NHS chemistry. Two variants of high-affinity binding molecules were used to detect target molecule. Firstly, a variant is based on ABD domain fused with TolA chain. This molecule is in vivo biotinylated and each molecule contains one molecule of biotin and one ABD domain. Secondly, the variant is ABD domain based on streptavidin molecule and contains four gaps for biotin and four ABD domains. These high-affinity molecules were immobilized to the chip surface via biotin-streptavidin chemistry. To eliminate nonspecific binding 1% bovine serum albumin (BSA) or 6% fetal bovine serum (FBS) were used in every step. For both variants range of measured concentrations of fluorescently labelled HSA was 0 – 30 µg/ml. As a control, we performed a simultaneous assay without high-affinity binding molecules. Fluorescent signal was measured using inverse fluorescent microscope Olympus IX 70 with COOL LED pE 4000 as a light source, related filters, and camera Retiga 2000R as a detector. The fluorescent signal from non-modified areas was substracted from the signal of the fluorescent areas. Results were presented in graphs showing the dependence of measured grayscale value on the log-scale of HSA concentration. For the TolA variant the limit of detection (LOD) of the optical immunosensor proposed in this study is calculated to be 0,20 µg/ml for HSA detection in 1% BSA and 0,24 µg/ml in 6% FBS. In the case of streptavidin-based molecule, it was 0,04 µg/ml and 0,07 µg/ml respectively. The dynamical range of the immunosensor was possible to estimate just in the case of TolA variant and it was calculated to be 0,49 – 3,75 µg/ml and 0,73-1,88 µg/ml respectively. In the case of the streptavidin-based the variant we didn´t reach the surface saturation even with the 480 ug/ml concentration and the upper value of dynamical range was not estimated. Lower value was calculated to be 0,14 µg/ml and 0,17 µg/ml respectively. Based on the obtained results, it´s clear that both variants are useful for creating the bio-recognizing layer on immunosensors. For this particular system, it is obvious that the variant based on streptavidin molecule is more useful for biosensing on glass planar surfaces. Immunosensors based on this variant would exhibit better limit of detection and wide dynamical range.

Keywords: high affinity binding molecules, human serum albumin, optical immunosensor, protein G, UV-photolitography

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Authors: Alena Cejkova, Martina Paldrychova, Jana Michailidu, Olga Matatkova, Jan Masak

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Increasing the resistance of pathogenic microorganisms to many antibiotics is a serious threat to the treatment of infectious diseases and cleaning medical instruments. It should be added that the resistance of microbial populations growing in biofilms is often up to 1000 times higher compared to planktonic cells. Biofilm formation in a number of microorganisms is largely influenced by the quorum sensing regulatory mechanism. Finding external factors such as natural substances or physical processes that can interfere effectively with quorum sensing signal molecules should reduce the ability of the cell population to form biofilm and increase the effectiveness of antibiotics. The present work is devoted to the effect of chitosan as a representative of natural substances with anti-biofilm activity and non- thermal plasma (NTP) alone or in combination with polymyxin B on biofilm formation of Pseudomonas aeruginosa. Particular attention was paid to the influence of these agents on the level of quorum sensing signal molecules (acyl-homoserine lactones) during planktonic and biofilm cultivations. Opportunistic pathogenic strains of Pseudomonas aeruginosa (DBM 3081, DBM 3777, ATCC 10145, ATCC 15442) were used as model microorganisms. Cultivations of planktonic and biofilm populations in 96-well microtiter plates on horizontal shaker were used for determination of antibiotic and anti-biofilm activity of chitosan and polymyxin B. Biofilm-growing cells on titanium alloy, which is used for preparation of joint replacement, were exposed to non-thermal plasma generated by cometary corona with a metallic grid for 15 and 30 minutes. Cultivation followed in fresh LB medium with or without chitosan or polymyxin B for next 24 h. Biofilms were quantified by crystal violet assay. Metabolic activity of the cells in biofilm was measured using MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) colorimetric test based on the reduction of MTT into formazan by the dehydrogenase system of living cells. Activity of N-acyl homoserine lactones (AHLs) compounds involved in the regulation of biofilm formation was determined using Agrobacterium tumefaciens strain harboring a traG::lacZ/traR reporter gene responsive to AHLs. The experiments showed that both chitosan and non-thermal plasma reduce the AHLs level and thus the biofilm formation and stability. The effectiveness of both agents was somewhat strain dependent. During the eradication of P. aeruginosa DBM 3081 biofilm on titanium alloy induced by chitosan (45 mg / l) there was an 80% decrease in AHLs. Applying chitosan or NTP on the P. aeruginosa DBM 3777 biofilm did not cause a significant decrease in AHLs, however, in combination with both (chitosan 55 mg / l and NTP 30 min), resulted in a 70% decrease in AHLs. Combined application of NTP and polymyxin B allowed reduce antibiotic concentration to achieve the same level of AHLs inhibition in P. aeruginosa ATCC 15442. The results shown that non-thermal plasma and chitosan have considerable potential for the eradication of highly resistant P. aeruginosa biofilms, for example on medical instruments or joint implants.

Keywords: anti-biofilm activity, chitosan, non-thermal plasma, opportunistic pathogens

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Authors: Alena Nesterenko, Svetlana Petrikova

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Research evaluation is one of the most important elements of self-regulation and development of researchers as it is impartial and independent process of assessment. The method of expert evaluations as a scientific instrument solving complicated non-formalized problems is firstly a scientifically sound way to conduct the assessment which maximum effectiveness of work at every step and secondly the usage of quantitative methods for evaluation, assessment of expert opinion and collective processing of the results. These two features distinguish the method of expert evaluations from long-known expertise widespread in many areas of knowledge. Different typical problems require different types of expert evaluations methods. Several issues which arise with these methods are experts’ selection, management of assessment procedure, proceeding of the results and remuneration for the experts. To address these issues an on-line system was created with the primary purpose of development of a versatile application for many workgroups with matching approaches to scientific work management. Online documentation assessment and statistics system allows: - To realize within one platform independent activities of different workgroups (e.g. expert officers, managers). - To establish different workspaces for corresponding workgroups where custom users database can be created according to particular needs. - To form for each workgroup required output documents. - To configure information gathering for each workgroup (forms of assessment, tests, inventories). - To create and operate personal databases of remote users. - To set up automatic notification through e-mail. The next stage is development of quantitative and qualitative criteria to form a database of experts. The inventory was made so that the experts may not only submit their personal data, place of work and scientific degree but also keywords according to their expertise, academic interests, ORCID, Researcher ID, SPIN-code RSCI, Scopus AuthorID, knowledge of languages, primary scientific publications. For each project, competition assessments are processed in accordance to ordering party demands in forms of apprised inventories, commentaries (50-250 characters) and overall review (1500 characters) in which expert states the absence of conflict of interest. Evaluation is conducted as follows: as applications are added to database expert officer selects experts, generally, two persons per application. Experts are selected according to the keywords; this method proved to be good unlike the OECD classifier. The last stage: the choice of the experts is approved by the supervisor, the e-mails are sent to the experts with invitation to assess the project. An expert supervisor is controlling experts writing reports for all formalities to be in place (time-frame, propriety, correspondence). If the difference in assessment exceeds four points, the third evaluation is appointed. As the expert finishes work on his expert opinion, system shows contract marked ‘new’, managers commence with the contract and the expert gets e-mail that the contract is formed and ready to be signed. All formalities are concluded and the expert gets remuneration for his work. The specificity of interaction of the examination officer with other experts will be presented in the report.

Keywords: expertise, management of research evaluation, method of expert evaluations, research evaluation

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