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Úloha Adenylát kinázy 1 v aktivaci a metabolismu imunitních buněk larev \kur{Drosophila melanogaster}
KAISLEROVÁ, Nikola
The aim of this thesis was to study the role of Adenylate kinase 1 (Ak1) in the immune system of Drosophila melanogaster larvae upon the infection by parasitoid wasp Leptopilina boulardi. Using the immune specific Ak1 RNA interference, it was analyzed the effect of Ak1 reduction on the immune response and viability of Drosophila. The importance of Ak1 was also evaluated within the metabolism of immune cells. It has been shown that Ak1 is crucial in energy metabolism of immune cells and important for the proper functioning of immune system.
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Effects of natural compounds on viability of tumor cell lines
Boleslavská, Barbora ; Poljaková, Jitka (advisor) ; Bořek Dohalská, Lucie (referee)
Cancer is considered to be one of the most serious issues of medicine nowadays. Moreover, its incidence is still rising. Despite the huge progress in modern treatment methods, cancer therapy is still limited by many difficulties. This work focuses on the natural substances such as epigallocatechin gallate, caffeine, Cannabis sativa ethanol extract, Origanum acutidens water extract, Mentha piperita water extract and its effects on the human neuroblastoma cell line UKF-NB-4. The first part of the bachelor thesis deals with determining the viability of human neuroblastoma cell line UKF-NB-4 exposed to tested substances as well as it studies the effects of those substances on the cell cycle and caspase activity. Finally, it was tested, whether those substances are able to induce apoptosis in neuroblastoma cell lines. Tests were undertaken on the MUSETM cell analyzer and on the flow cytometer. The second part of the bachelor thesis focuses on the expression of protein p53 and retinoblastoma protein in neuroblastoma cell lines exposed to tested substances. Detection was carried out by Western blot analysis. Epigallocatechin gallate exhibited the most significant effect on human neuroblastoma cell line. It lowers the expression of retinoblastoma protein as well as it arrests cells between G0/G1 and S...
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Ecophysiology of microscopic fungus Pseudogymnoascus destructans
Homutová, Karolína ; Kubátová, Alena (advisor) ; Baldrian, Petr (referee)
A microscopic fungus Pseudogymnoascus destructans (Ascomycota: Pseudeurotiaceae) causes illness known as white-nose syndrome (WNS) causing death of bats during hibernation. The illness occurs in the North America and in Europe. The fungus is characteristic by asymmetrically curved conidia, by slow growth and growth at low temperatures (below 20 řC). The aim of this study is to clarify properties responsible for unique ecelogy of Pseudogymnoascus destructans by comparison with ecological related or unrelated pathogenic or nonpathogenic fungi. This part includes study of tolerance to physiological stresses and recognition of spectrum of utilizating nutrients (compounds of carbon, nitrogen, phosphorus, sulphur and nutrient supplements). Testing to physiological stresses should help to estimate a potentiality of fungus to spread out of caves. The last aim is to develop a selective isolation medium for P. destructans. Influence of several types of physiological stress (e.g. UVA, UVA with UVB, 25 řC, 30 řC, 37 řC and dryness) was investigated with fluorescent stain propidium iodide (PI) by flow cytometry. The spores of Pseudogymnoascus destructans and three fungi from underground spaces were not viable after 3 weeks at 37 řC. Other stresses did not cause a decreasing of viability or some stresses caused...
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Comparison of antimicrobial effect of emulsions for cleaning and dissinfection
Chrápavá, Edita ; Dzurická, Lucia (referee) ; Márová, Ivana (advisor)
Currently, hand disinfectants and other disinfectants are in high demand. Due to the pandemic and the spread of Covid-19, they are used by society daily. The reason for their use is not only to prevent the spread of bacteria and other microorganisms from the hands to another part of the body or objects, but also to prevent the spread of the disease. The subject of the bachelor thesis is the control of antimicrobial effects of several commonly available disinfectants using broth and agar dilution methods. From the measured data, the absorbance was processed and subsequently viability of the culture.
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Changes of microbial growth during use of face masks from different materials
Korvasová, Lucie ; Hoová, Julie (referee) ; Márová, Ivana (advisor)
Due to the current spread of Covid-19 disease, there are many types of respiratory protective equipment on the Czech market. However, due to the conditions that breath creates on the inside of the masks, they can be an ideal place for multiplication of microorganisms such as yeasts, moulds and especially bacteria. Since various protection masks differ in their production materials, bacteria colonization won’t always be the same. The subject of the thesis is a comparison of individual types of masks and respirators with regard to multiplication of microorganisms in their structure, which were tested using the contact stamp-form method on agar plates and in a liquid medium. Sterile environment was required during the work to avoid external contamination. Methods of imprinting on agar plates and the fact that turbidity and thus measurable absorbance increases with rising number of microorganisms were used to obtain the data. Result of the work was that face masks already contained some level of contamination from their production. As the best kind of face masks in terms of multiplication of microorganisms came out cotton, which prevented multiplying in the inoculated sphere. At the same time each growing culture was tested for viability, where it was determined how many viable cells each sample contained in comparison to a fully grown culture.
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Addition of probiotics and algae extracts to baby foods
Liová, Alexandra ; Szotkowski, Martin (referee) ; Němcová, Andrea (advisor)
This bachelor thesis deals with the selection of a suitable combination of probiotic culture with algae extracts as an addition to the baby formula and the application of the optimal composition of baby formula. Arame, wakame, hijiki and kombu seaweeds were selected for these purposes. The theoretical part is focused on the proper nutrition of children, their deficiencies and possible diseases. Subsequently, it deals with the characterization of probiotic cultures and their importance in the child's nutrition. Last but not least, the theoretical part characterizes the individual seaweeds, their nutritional composition and also their importance in the child's nutrition. In the experimental part, samples of selected algae were first analyzed in terms of saccharides, insoluble fiber content, fructooligosaccharides and -glucans or pigment content. A sample of inulin, a commonly used prebiotic in probiotic products, was used as a reference component. An important analysis of this experimental part was to determine the effect of algal extracts on the viability and growth of probiotic bacteria. Subsequently, a model digestion of a mixture of probiotics and algal extracts was performed, and the condition of the probiotic cells was evaluated during digestion and after digestion by measuring turbidity and flow cytometry. Finally, not only the optimal combination of algae extract and probiotic culture was proposed, but also the possibility of improving the long-term viability of probiotic cells in combination with algal extracts.
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Study on PHa biosynthesis by cyanobacteria
Drinka, Jakub ; Kouřilová, Xenie (referee) ; Obruča, Stanislav (advisor)
This bachelor thesis concerns with the effect of selected environmental stresses towards the viability of cyanobacterial species Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. Both strains were studied via flow cytometry using the appropriate fluorochromes while also observing the applicability of these stains and precision of this method in the analysis of cyanobacterial cultures. In spite of the optimization being carried out, fluorescein diacetate was not suitable for the analysis of these cells because of their autofluorescence interfere with the fluorochrome emissions. Therefore, SYTOXTM Blue Dead Cell stain was used for the further experiments. A cryoprotective effect of polyhydroxybutyrate was observed during the freezing-thawing cycles. On the other hand, the culture rich in PHB seemed to be less resistant towards the oxidative stress induced by the hydrogen peroxide, probably justifiable by the state of chlorosis. Furthermore, this thesis deals with the increase/decrease in biomass concentration and intracellular PHB pool as a response to the external carbon source addition, analysed by the gas chromatography paired with FID detection. The most intesive cell growth in Synechocystis sp. PCC 6803 was detected in the presence of sodium acetate combined with gamma-butyrolactone while the highest PHB content in the medium supplied with acetate and 1,4-butanediol. Nevertheless, Synechocystis salina CCALA 192 showed markedly higher polyhydroxybutyrate accumulation under both photoautotrophic and mixotrophic conditions, the only non-suitable substrate was gamma-butyrolactone.
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Utilization of physicochemical and spectroscopic techniques in study of stress-response of cyanobacteria
Skoryk, Maksym ; Šedrlová, Zuzana (referee) ; Sedláček, Petr (advisor)
Tato bakalářská práce je soustředěná na zkoumaní cyanobakterií, vystavených hyper a hyposmotickým stresům. Na základě literární rešerše byly navžené vhodné analytické metody určené ke studiu dvou modelových organismu - Synechocystis sp. PCC 6803 a Synechocystis sp. salina Wislouch CCALA 192. Celkem čtyři metody byly použité k charakterizaci osmoticky zatížených bakterií. Průtoková cytometrie byla použita k vyhodnocení viability. Fluorescenční sonda SYTOX Blue poskytla důvěryhodnou informaci ohledně viability bakterií. Propidium jodid naopak poskytl nezřetelnou informaci. Optické vlastnosti cyanobakterií byly prozkoumané pomoci UV-VIS absorpčních a turbidimetrických měření. Termogravimetrická analýza byla použita pro mapování změn obsahu vody v osmoticky stresovaných buňkách. Tato metoda ukazala. že PHB-positivní Synechocystis sp. PCC 6803 jsou pravděpodobně vice odolné vůči hyperosmotickým stresům než PHB-negativní. Plynová chromatografie byla použita ke kvantifikaci vnitrobuněčného PHB, který činil přibližně 1-2 % suché hmoty PHB-positivních buněk.
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Influence of PHA accumulation on resistance of bacteria against selected antibacterial drugs
Hrabalová, Vendula ; Kučera, Dan (referee) ; Obruča, Stanislav (advisor)
The aim of this bachelor thesis was to study the effect of bactericidal drugs on bacteria from the genus C. necator H16 and its mutant genus PHB-4. The genus H16 shows ability to accumulate polyhydroxyalkanoates (PHA) in the form of granules while the genus PHB-4 lacks to show this ability. The theoretical part of the bachelor thesis is focused on the effect of antibiotics on bacteria in general and the determination of susceptibility of bacteria to antimicrobial substances. The effect of three specific antibiotics (nisine, streptomycin and penicillin) on both bacterial strains was tested in the experimental part. The viability of bacteria was determined by the spread plate method and flow cytometry. Agar diffusion test and broth microdilution test were used to test the susceptibility of bacteria. It was concluded that the accumulation of PHA decreases the tolerance of bacteria to antimicrobial substances because the genus C. necator H16 is more susceptible to streptomycin and penicillin then the strain C. necator PHB-4.
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Biological effects of substances isolated from Isoptera species
Dušková, Simona ; Hrstka, Miroslav (referee) ; Brázda, Václav (advisor)
This thesis was focused on monitoring the viability of eukaryotic and prokaryotic cells after exposure of termites-isolated chemicals. Recently, evidence of antibacterial and antifungal properties of these defense substances has grown, and they can find a wide range of uses not only in the pharmaceutical industry. In this work, three defensive substances from termites were studied: nerolidol, nitropentadecene and methylanthranilate. Their antibacterial effects, minimal inhibitory concentrations and minimal bactericidal concentrations against Escherichia coli STBL3 strain were monitored. Further, their cytotoxic effects on eukaryotic non-tumor (HEK293FT) and tumor cells (MCF7) as well as their effect on plasmid DNA were studied. Antibiotic ampicillin and cytostatic cisplatin were used as control substances for antibacterial and cytotoxic effects, respectively. In the case of the action of nerolidol, nitropentadecene and methylanthranilate on the STBL3 strain, antibacterial activity was not demonstrated. Cytotoxic effects were observed nerolidol and nitropentadecene. None of the examined substances modified the plasmid DNA.
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