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Gold nano systems for the detection of molecules using surface-enhanced Raman scattering (SERS)
Benešová, Markéta ; Bernatová,, Silvie (referee) ; Skoumalová, Petra (advisor)
This diploma thesis was focused on the production and use of gold nanosystems to enhance Raman scattering. Metal nanoparticles, when interacting with electromagnetic radiation, form so-called localized plasmons, which can enhance Raman scattering. SERS (surface-enhanced Raman scattering) is a non-destructive analytical technique used in this work to measure the concentration and chemical changes in the rhodamine B molecule. Rhodamine B was subjected to photocatalytic degradation using two types of photocatalysts before measurement: TiO2-(H) and TiO2-(H)-Ag. The quantitative dependence of rhodamine B photodegradation on the presence of photocatalysts in a given period time was sought using photodegradation processes. The results of measurements using the SERS method were compared with the results obtained by UV-VIS spectroscopy. From the measured data, it was found that the photocatalysts significantly accelerate the photodegradation processes, because the Raman signal of rhodamine B decreased, while the signal decrease was most pronounced for the catalyst with added TiO2-(H)-Ag, less prominent but still statistically significant signal decrease was observed for the TiO2-(H) catalyst. In the control sample without the addition of photocatalyst, no decrease in signal was observed. In the next phase of the thesis, a sandwich immunoassay was designed that uses SERS to detect E. coli bacteria or other specific microorganisms in the sample. The first component of the sandwich immunoassay is gold nanoparticles, which carry a so-called Raman reporter, which has a clear Raman response in the spectrum, and gold nanoparticles amplify this signal, and antibodies, thanks to which the particles specifically bind to the microorganism. Another component is either gold layered slides or magnetic nanoparticles, which are modified with antibodies and serve to immobilize microorganisms. This system can be a fast and very accurate way to identify a given microorganism in a sample.
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Determination of drugs by HPLC with different detectors
Benešová, Markéta ; Opatřilová, Radka (referee) ; Vávrová, Milada (advisor)
This diploma thesis deals with the determination of macrolide antibiotics in wastewater, especially with erythromycin, clarithromycin and roxithromycin. In this time are these pharmaceuticals prescribed quite frequently. Solid phase extraction (SPE) was used for the isolation and the purification of selected analytes from an aqueous matrix; as the suitable procedure was found the using Oasis HLB cartridges. High performance liquid chromatography with mass spectrometry detection (HPLC-MS) was optimized for its analysis of selected pharmaceuticals. The optimized method was used for the determination of pharmaceuticals in real water samples, which was taken at the inflow and the outflow of the urban wastewater treatment plant in Brno-Modřice.
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Question of determination of analgetics in waters
Benešová, Markéta ; Lisá, Hana (referee) ; Vávrová, Milada (advisor)
The bachelor thesis is focused on the optimization of the method for the determination of analgetics in waters. The aim of this work is to work up the methodical procedure for determination of chosen pharmaceuticals i.e. salicylic acid and naproxen in model water samples. Analytes were isolated by solid phase extraction (SPE); they were identified by high performance liquid chromatography with UV-VIS detection of diode – array type (HPLC – DAD). The optimal method was selected from several verified procedures on the basis of practical experience.
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Preparation of antimicrobial gels with plant extract and oils
Benešová, Markéta ; Hoová, Julie (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis was focused on studying antimicrobial effects of natural plant extracts and their subsequent application. The research was mainly focused on the antioxidant and antimicrobial activity of plant extracts and liposomal particles with their contents. Theoretical part of thesis was focused on the topic of natural antimicrobial and antioxidant substances, problems with bacterial resistance as well as describing features and attributes of used herbs, and characterization of gels and liposomes. Within the experimental part of thesis, three types of extracts (aquatic, ethanol and oil), prepared from 10 different types of plants, were described. The content of phenolic, flavonoid substances, also antioxidant and antimicrobial activity were then determined. After that, the contents of all extracts were encapsulated into liposomes, and size, stability, encapsulation activity, also antioxidant and antimicrobial activity of those liposomes were determined. The size distribution was wide, but in terms of the measured zeta potential, most of the particles were stable. All prepared extracts and liposomes containing them were tested for their antimicrobial activity. The tests were performed on a gram-positive genus of Micrococcus luteus and a gram-negative type of Serratia marcescens. The most effective oil extracts against both species were eucalyptus and fir, as well as oil extract of cloves, along with aqueous extracts of mustard seeds, cloves and juniper, or ethanol extract of anise which also has high antimicrobial effects. Finally, these extracts and liposomes were applied into the gels, which could be found useful in the pharmaceutical industry, as an alternative to externally used antibiotics, or in the cosmetic industry.
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SERS-TAGS: Selective immobilization and detection of bacteria using specific antibodies and surface-enhanced Raman scattering
Benešová, Markéta ; Bernatová, Silvie ; Samek, Ota ; Pokorná, Zuzana ; Mika, Filip ; Kizovský, Martin ; Pilát, Zdeněk
Raman spectroscopy is a non-destructive instrumental analytical technique based on Raman scattering, which is the inelastic scattering of photons that occurs when interacting with electrons in chemical bonds. With the help of Raman spectroscopy, chemical compounds, their mixtures and biological samples, including living organisms, can be analyzed very quickly, non-contactly and non-destructively. The performed measurements can be compared with spectra databases. A problem with Raman spectroscopy is that samples sometimes give a weak signal that is often overlaid by intense fluorescence. SERS (surface-enhanced Raman spectroscopy) is used to amplify Raman scattering.
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Everydayness in teaching regional geography
Benešová, Markéta ; Matějček, Tomáš (advisor) ; Cimová, Tereza (referee)
The aim of the diploma thesis is to find out the potential of using the phenomenon of the everydayness in the teaching of regional geography. In the theoretical part, the concept of everydayness is presented from different perspectives, and we further focus on its connection with geography. In the practical part of the diploma thesis, several activities based on the stories of the daily life of the inhabitants of the selected region are put forward and tested in the lesson. The investigation was carried out using qualitative research methods. The source of the data were the notes from the participants' observation conducted during the lesson and the submitted outputs of the students (in the form of worksheets or a short questionnaire). These data were then analyzed, and the results were subjected to a discussion in which they were confronted with the fulfillment of the respective objectives. The conclusion of the thesis contains, among other things, suggestions for further possible geographical research in the field of everydayness. key words: everydayness, regional geography, geography of everyday life, geographic education, qualitative research
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Gold nanosystems for the detection of molecules using surface-enhanced Raman scatterings (SERS)
Benešová, Markéta
Raman spectroscopy is a non-destructive analytical technique to analyze the chemical structure of molecules by a phenomenon known as Raman scattering, which occurs by an inelastic interaction of photons with the valence electrons in molecular bonds. However, Raman scattering can be hard to observe due to other, more frequent phenomena, such as Rayleigh scattering or fluorescence. SERS (surface-enhanced Raman spectroscopy) uses localized surface plasmon resonance (LSPR) of metal nanostructures to amplify Raman scattering. LSPR is a coherent oscillation of conduction electrons that arises from the interaction of electromagnetic radiation with metal nanostructures. The amplification of Raman scattering occurs when the analyte is adsorbed on the surface of such nanostructure and the strong localized electric field interacts with the electrons in its molecular bonds. Signal amplification of several orders of magnitude can be achieved, commonly 103 or more. In our work, we determined the presence of a selected bacterial species by multi-functionalized golden nanoparticles called SERS-tags, which have their surface modified with an antibody and a Raman reporter. The antibody allows the nanoparticles to bind to the surface of a concrete bacterial species based on the antigen-antibody affinity. When the targeted bacterium is covered with the nanoparticles, the Raman reporter signal is amplified by SERS, providing specific and strong Raman response. Therefore, when the Raman reporter signal is detected in a sample, it confirms the presence of the specific bacterium on a single-cell level.
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Gold nano systems for the detection of molecules using surface-enhanced Raman scattering (SERS)
Benešová, Markéta ; Bernatová,, Silvie (referee) ; Skoumalová, Petra (advisor)
This diploma thesis was focused on the production and use of gold nanosystems to enhance Raman scattering. Metal nanoparticles, when interacting with electromagnetic radiation, form so-called localized plasmons, which can enhance Raman scattering. SERS (surface-enhanced Raman scattering) is a non-destructive analytical technique used in this work to measure the concentration and chemical changes in the rhodamine B molecule. Rhodamine B was subjected to photocatalytic degradation using two types of photocatalysts before measurement: TiO2-(H) and TiO2-(H)-Ag. The quantitative dependence of rhodamine B photodegradation on the presence of photocatalysts in a given period time was sought using photodegradation processes. The results of measurements using the SERS method were compared with the results obtained by UV-VIS spectroscopy. From the measured data, it was found that the photocatalysts significantly accelerate the photodegradation processes, because the Raman signal of rhodamine B decreased, while the signal decrease was most pronounced for the catalyst with added TiO2-(H)-Ag, less prominent but still statistically significant signal decrease was observed for the TiO2-(H) catalyst. In the control sample without the addition of photocatalyst, no decrease in signal was observed. In the next phase of the thesis, a sandwich immunoassay was designed that uses SERS to detect E. coli bacteria or other specific microorganisms in the sample. The first component of the sandwich immunoassay is gold nanoparticles, which carry a so-called Raman reporter, which has a clear Raman response in the spectrum, and gold nanoparticles amplify this signal, and antibodies, thanks to which the particles specifically bind to the microorganism. Another component is either gold layered slides or magnetic nanoparticles, which are modified with antibodies and serve to immobilize microorganisms. This system can be a fast and very accurate way to identify a given microorganism in a sample.
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Conservation genetics of grey wolf and snow leopard: effect of landscape attributes to the population structure
Benešová, Markéta ; Hulva, Pavel (advisor) ; Kreisinger, Jakub (referee)
Landscape genetic approaches allow to study effects of landscape to population microevolution. Landscape can influence gene flow even in large carnivores with good dispersal ability. Understanding the influence of landscape to the gene flow between populations is crucial for species conservation, especially in the species with low population densities. Aim of the study was to describe genetic structure of the grey wolf (Canis lupus) and snow leopard (Panthera uncia) in selected areas and to determine the influence of the landscape features on observed structure. Non-invasive genetic samples of snow leopard from Nepal were analysed, as well as invasive and non-invasive samples of grey wolf from Central Europe. Population structure was determined a posteriori using Bayesian clustering approaches that integrate genetic and geographical data, and compared to landscape connectivity models. Population structure of snow leopards is mostly influenced by human presence and presence of frequented roads, which represent a substantial dispersal barrier. Habitat suitable for this species is greatly restricted by altitude, however, during dispersal they are able to overcome areas with higher elevation than what is optimal for them. Pronounced genetic difference was found between central European and Carpathian...
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Preparation of antimicrobial gels with plant extract and oils
Benešová, Markéta ; Hoová, Julie (referee) ; Skoumalová, Petra (advisor)
This bachelor thesis was focused on studying antimicrobial effects of natural plant extracts and their subsequent application. The research was mainly focused on the antioxidant and antimicrobial activity of plant extracts and liposomal particles with their contents. Theoretical part of thesis was focused on the topic of natural antimicrobial and antioxidant substances, problems with bacterial resistance as well as describing features and attributes of used herbs, and characterization of gels and liposomes. Within the experimental part of thesis, three types of extracts (aquatic, ethanol and oil), prepared from 10 different types of plants, were described. The content of phenolic, flavonoid substances, also antioxidant and antimicrobial activity were then determined. After that, the contents of all extracts were encapsulated into liposomes, and size, stability, encapsulation activity, also antioxidant and antimicrobial activity of those liposomes were determined. The size distribution was wide, but in terms of the measured zeta potential, most of the particles were stable. All prepared extracts and liposomes containing them were tested for their antimicrobial activity. The tests were performed on a gram-positive genus of Micrococcus luteus and a gram-negative type of Serratia marcescens. The most effective oil extracts against both species were eucalyptus and fir, as well as oil extract of cloves, along with aqueous extracts of mustard seeds, cloves and juniper, or ethanol extract of anise which also has high antimicrobial effects. Finally, these extracts and liposomes were applied into the gels, which could be found useful in the pharmaceutical industry, as an alternative to externally used antibiotics, or in the cosmetic industry.
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