Search results for: Radka Obořilová
8 Advanced Biosensor Characterization of Phage-Mediated Lysis in Real-Time and under Native Conditions
Authors: Radka Obořilová, Hana Šimečková, Matěj Pastucha, Jan Přibyl, Petr Skládal, Ivana Mašlaňová, Zdeněk Farka
Abstract:
Due to the spreading of antimicrobial resistance, alternative approaches to combat superinfections are being sought, both in the field of lysing agents and methods for studying bacterial lysis. A suitable alternative to antibiotics is phage therapy and enzybiotics, for which it is also necessary to study the mechanism of their action. Biosensor-based techniques allow rapid detection of pathogens in real time, verification of sensitivity to commonly used antimicrobial agents, and selection of suitable lysis agents. The detection of lysis takes place on the surface of the biosensor with immobilized bacteria, which has the potential to be used to study biofilms. An example of such a biosensor is surface plasmon resonance (SPR), which records the kinetics of bacterial lysis based on a change in the resonance angle. The bacteria are immobilized on the surface of the SPR chip, and the action of phage as the mass loss is monitored after a typical lytic cycle delay. Atomic force microscopy (AFM) is a technique for imaging of samples on the surface. In contrast to electron microscopy, it has the advantage of real-time imaging in the native conditions of the nutrient medium. In our case, Staphylococcus aureus was lysed using the enzyme lysostaphin and phage P68 from the familyPodoviridae at 37 ° C. In addition to visualization, AFM was used to study changes in mechanical properties during lysis, which resulted in a reduction of Young’s modulus (E) after disruption of the bacterial wall. Changes in E reflect the stiffness of the bacterium. These advanced methods provide deeper insight into bacterial lysis and can help to fight against bacterial diseases.Keywords: biosensors, atomic force microscopy, surface plasmon resonance, bacterial lysis, staphylococcus aureus, phage P68
Procedia PDF Downloads 1347 Chloride Transport in Ultra High Performance Concrete
Authors: Radka Pernicova
Abstract:
Chloride resistance in Ultra High Performance Concrete (UHPC) is determined in this paper. This work deals with the one dimension chloride transport, which can be potentially dangerous particularly for the durability of concrete structures. Risk of reinforcement corrosion due to exposure to the concrete surface to direct the action of chloride ions (mainly in the form de-icing salts or groundwater) is dangerously increases. The measured data are investigated depending on the depth of penetration of chloride ions into the concrete structure. Comparative measurements with normal strength concrete are done as well. The experimental results showed that UHCP have improved resistance of chlorides penetration than NSC and also chloride diffusion depth is significantly lower in UHCP.Keywords: chloride, one dimensional diffusion, transport, salinity, UHPC
Procedia PDF Downloads 4346 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
Abstract:
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
Procedia PDF Downloads 3705 Potential of Salvia sclarea L. for Phytoremediation of Soils Contaminated with Heavy Metals
Authors: Violina R. Angelova, Radka V. Ivanova, Givko M. Todorov, Krasimir I. Ivanov
Abstract:
A field study was conducted to evaluate the efficacy of Salvia sclarea L. for phytoremediation of contaminated soils. The experiment was performed on an agricultural fields contaminated by the Non-Ferrous-Metal Works near Plovdiv, Bulgaria. The content of heavy metals in different parts of Salvia sclarea L. (roots, stems, leaves and inflorescences) was determined by ICP. The essential oil of the Salvia sclarea L. was obtained by steam distillation in laboratory conditions and was analyzed for heavy metals and its chemical composition was determined. Salvia sclarea L. is a plant which is tolerant to heavy metals and can be grown on contaminated soils. Based on the obtained results and using the most common criteria, Salvia sclarea L. can be classified as Pb hyperaccumulator and Cd and Zn accumulators, therefore, this plant has suitable potential for the phytoremediation of heavy metal contaminated soils. Favorable is also the fact that heavy metals do not influence the development of the Salvia sclarea L., as well as on the quality and quantity of the essential oil. For clary sage oil obtained from the processing of clary sage grown on highly contaminated soils, its key odour-determining ingredients meet the quality requirements of the European Pharmacopoeia and BS ISO 7609 regarding Bulgarian clary sage oil and/or have values that are close to the limits of these standards. The possibility of further industrial processing will make Salvia sclarea L. an economically interesting crop for farmers of phytoextraction technology.Keywords: clary sage, heavy metals, phytoremediation, polluted soils
Procedia PDF Downloads 2164 Effect of Compost Application on Uptake and Allocation of Heavy Metals and Plant Nutrients and Quality of Oriental Tobacco Krumovgrad 90
Authors: Violina R. Angelova, Venelina T. Popova, Radka V. Ivanova, Givko T. Ivanov, Krasimir I. Ivanov
Abstract:
A comparative research on the impact of compost on uptake and allocation of nutrients and heavy metals and quality of Oriental tobacco Krumovgrad 90 has been carried out. The experiment was performed on an agricultural field contaminated by the lead zinc smelter near the town of Kardzali, Bulgaria, after closing the lead production. The compost treatments had significant effects on the uptake and allocation of plant nutrients and heavy metals. The incorporation of compost leads to decrease in the amount of heavy metals present in the tobacco leaves, with Cd, Pb and Zn having values of 36%, 12% and 6%, respectively. Application of the compost leads to increased content of potassium, calcium and magnesium in the leaves of tobacco, and therefore, may favorably affect the burning properties of tobacco. The incorporation of compost in the soil has a negative impact on the quality and typicality of the oriental tobacco variety of Krumovgrad 90. The incorporation of compost leads to an increase in the size of the tobacco plant leaves, the leaves become darker in colour, less fleshy and undergo a change in form, becoming (much) broader in the second, third and fourth stalk position. This is accompanied by a decrease in the quality of the tobacco. The incorporation of compost also results in an increase in the mineral substances (pure ash), total nicotine and nitrogen, and a reduction in the amount of reducing sugars, which causes the quality of the tobacco leaves to deteriorate (particularly in the third and fourth harvests).Keywords: chemical composition, compost, heavy metals, oriental tobacco, quality
Procedia PDF Downloads 2733 Non-Invasive Techniques of Analysis of Painting in Forensic Fields
Authors: Radka Sefcu, Vaclava Antuskova, Ivana Turkova
Abstract:
A growing market with modern artworks of a high price leads to the creation and selling of artwork counterfeits. Material analysis is an important part of the process of assessment of authenticity. Knowledge of materials and techniques used by original authors is also necessary. The contribution presents possibilities of non-invasive methods of structural analysis in research on paintings. It was proved that unambiguous identification of many art materials is feasible without sampling. The combination of Raman spectroscopy with FTIR-external reflection enabled the identification of pigments and binders on selected artworks of prominent Czech painters from the first half of the 20th century – Josef Čapek, Emil Filla, Václav Špála and Jan Zrzavý. Raman spectroscopy confirmed the presence of a wide range of white pigments - lead white, zinc white, titanium white, barium white and also Freeman's white as a special white pigment of painting. Good results were obtained for red, blue and most of the yellow areas. Identification of green pigments was often impossible due to strong fluorescence. Oil was confirmed as a binding medium on most of the analyzed artworks via FTIR - external reflection. Collected data present the valuable background for the determination of art materials characteristic for each painter (his palette) and its development over time. Obtained results will further serve as comparative material for the authentication of artworks. This work has been financially supported by the project of the Ministry of the Interior of the Czech Republic: The Development of a Strategic Cluster for Effective Instrumental Technological Methods of Forensic Authentication of Modern Artworks (VJ01010004).Keywords: non-invasive analysis, Raman spectroscopy, FTIR-external reflection, forgeries
Procedia PDF Downloads 1722 Cytolethal Distending Toxins in Intestinal and Extraintestinal E. coli
Authors: Katarína Čurová, Leonard Siegfried, Radka Vargová, Marta Kmeťová, Vladimír Hrabovský
Abstract:
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
Procedia PDF Downloads 3501 Zinc Nanoparticles Modified Electrode as an Insulin Sensor
Authors: Radka Gorejova, Ivana Sisolakova, Jana Shepa, Frederika Chovancova, Renata Orinakova
Abstract:
Diabetes mellitus (DM) is a serious metabolic disease characterized by chronic hyperglycemia. Often, the symptoms are not sufficiently observable at early stages, and so hyperglycemia causes pathological and functional changes before the diagnosis of the DM. Therefore, the development of an electrochemical sensor that will be fast, accurate, and instrumentally undemanding is currently needful. Screen-printed carbon electrodes (SPCEs) can be considered as the most suitable matrix material for insulin sensors because of the small size of the working electrode. It leads to the analyst's volume reduction to only 50 µl for each measurement. The surface of bare SPCE was modified by a combination of chitosan, multi-walled carbon nanotubes (MWCNTs), and zinc nanoparticles (ZnNPs) to obtain better electrocatalytic activity towards insulin oxidation. ZnNPs were electrochemically deposited on the chitosan-MWCNTs/SPCE surface using the pulse deposition method. Thereafter, insulin was determined on the prepared electrode using chronoamperometry and electrochemical impedance spectroscopy (EIS). The chronoamperometric measurement was performed by adding a constant amount of insulin in 0.1 M NaOH and PBS (2 μl) with the concentration of 2 μM, and the current response of the system was monitored after a gradual increase in concentration. Subsequently, the limit of detection (LOD) of the prepared electrode was determined via the Randles-Ševčík equation. The LOD was 0.47 µM. Prepared electrodes were studied also as the impedimetric sensors for insulin determination. Therefore, various insulin concentrations were determined via EIS. Based on the performed measurements, the ZnNPs/chitosan-MWCNTs/SPCE can be considered as a potential candidate for novel electrochemical sensor for insulin determination. Acknowledgments: This work has been supported by the projects Visegradfund project number 22020140, VEGA 1/0095/21 of the Slovak Scientific Grant Agency, and APVV-PP-COVID-20-0036 of the Slovak Research and Development Agency.Keywords: zinc nanoparticles, insulin, chronoamperometry, electrochemical impedance spectroscopy
Procedia PDF Downloads 122