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Study of plasma jet effects on yeast
Trebulová, Kristína ; Skoumalová, Petra (referee) ; Krčma, František (advisor)
The purpose of this work is to study the effects of a 2.45 GHz microwave plasma jet on the yeast Candida glabrata. The microwave plasma was generated by a surfatron resonator and as a working gas pure argon at constant flow rate 5 Slm was used. Thanks to a high number of active particles generated in a low temperature plasma, this type of plasma has become highly popular, especially thanks to its bactericidal effects. However, its antifungal effects and mechanisms of fungal inactivation have still not been fully understood. Therefore, the study focuses on the antimycotic effects of the low temperature plasma in a form of a microwave discharge on Candida glabrata as a model yeast example. The principal focus is on the measurement and evaluation of changes in inactivation effects caused by varying the initial concentration of Candida glabrata cells, applied power, and the exposure time. The theoretical part presents the basic information concerning Candida glabrata, its pathogenicity, virulence and infectivity. Furthermore, plasma in general, various types of discharges and their applications in the food and biomedical industries are discussed. The experimental part focalizes on the determination of the inactivating effects of the microwave plasma jet on the yeast Candida glabrata. The experiment consists of applying the discharge on a freshly inoculated colony of Candida glabrata on agar plates and observing inhibitory effects in the form of inhibitory zones formed after subsequent cultivation. The inhibition of yeasts by the microwave discharge is interesting scientifically due to its promising results, which have also been proven in this work, but also economically, especially due to the use of argon. Argon as a carrier gas is much cheaper than helium and colder than common atmospheric pressure discharges in air such as DBDs, corona or glow discharges.
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Utilization of thermal analysis in the study on effects of microbial inhibitors
Bošeľová, Miriam ; Obruča, Stanislav (referee) ; Sedláček, Petr (advisor)
This diploma thesis deals with the use of thermal analysis in the study on effects of microbial inhibitors. The main aim of this work was to determine the utilization of the method, which is mainly used in different fields of science and research. Three bacterial strains: Cupriavidus necator H16, its mutant strain Cupriavidus necator PHB-4 and Halomonas halophila, were used as model microorganisms. The inhibitory effect of levulinic acid on growth and metabolic activity was monitored by microcalorimetry. It was found that bacteria were able to adapt to levulinic acid to a certain concentration - Cupriavidus necator to 5 g/l and Halomonas halophila to 2 g/l. The thermal analysis results were compared to a conventional method, which is commonly used to study the growth of microorganisms.
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Expression and binding properties of human galectin-4
Friede, Tereza ; Bojarová, Pavla (advisor) ; Kavan, Daniel (referee)
Galectin-4, a member of the galectin family known for its affinity to β-galactosides, belongs to tandem-repeat galectins and consists of two distinct subunits connected with a peptide linker. Galectins play a key role in various biological processes and are involved in the pathogenesis of many diseases. Colorectal cancer (CRC) tissue shows a lower expression of galectin-4 compared to healthy colon tissues, and the lower expression of galectin-4 promotes tumor progression and transport. Galectin-4 was found to inhibit tumorigenesis of CRC cells via Wnt/β-catenin and IL-6/NF-κB/STAT3 signaling pathways, which play a key role in CRC development. In inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, galectin-4 interacts with CD4+ T lymphocytes, influencing their behavior in the immunological synapsis. Furthermore, galectin-4, primarily expressed in the gastrointestinal tract, participates in the apical transport in epithelial cells, promotes the healing of intestinal epithelial wounds, participates in the adaptive immune response and has multiple roles in the central nervous system. Studying inhibitors of galectin-4 helps to understand its specific roles in these processes, and thus to identify new potential therapeutic targets. In this work human galectin-4 was heterologously...
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Studium změn antimikrobiální aktivity houby Ganoderma adspersum pomocí kultivační strategie OSMAC
Komárková, Lucie
Ganoderma adspersum is a white rot fungus that degrades lignocellulose in nature. However, it has also been proven to have many biological activities, including antioxidant, antitumor and anti-inflammatory. This study is focused on monitoring the antimicrobial activity of G. adspersum against the bacteria Escherichia coli, Staphylococcus aureus, MRSA and Klebsiella pneumoniae. First, the fungus was cultivated in three media with different additives. Using the minimum inhibitory concentration test, the effect of individual cultivation on viability was monitored. The lowest values were measured in YMB medium with tryptophan, which reduced the viability of E. coli up to 23 %. Based on this result, E. coli was chosen for growth curve, where media with additives were compared with pure media. It was found that pure media showed greater inhibitory activity than media with additives. The highest activity was observed in pure PDB medium, in which nearly 40% inhibition occurred at a concentration of 0,25 mg/ml. The results showed that Ganoderma adspersum could be a potential antimicrobial agent.
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Synthesis of peptides as potential ligands of aspartate protease HIV and MAY1 and confirmation of their inhibitory activity
Klikarová, Ivana ; Šácha, Pavel (advisor) ; Žáková, Lenka (referee)
The lentivirus known as the human immunodeficiency virus (HIV) is transmitted through blood and body fluids, causing the destruction of CD4 lymphocytes and leading to opportunistic infections that define acquired immunodeficiency syndrome (AIDS). The urgent need for new antiretroviral drugs stems from concerns about the long-term toxicity of existing drugs, HIV-1 variants resistant to treatment, and frequent changes in patient treatment. Drug development is focused on inhibitors of two viral enzymes, reverse transcriptase and protease. Antiretroviral therapy uses protease inhibitors in combination with nucleoside analogs to effectively suppress viral replication, prolonging the lives of HIV-infected patients and reducing morbidity. Cryptococcus neoformans and cryptococcus gattii infections primarily affect the immunocompromised population and have high morbidity and mortality rates. Resistance to commonly used antifungals has been emerging, making it more difficult to treat these infections. Protease inhibitor components used in antiretroviral therapies have shown some clinical efficacy in these opportunistic infections, particularly in Major Aspartyl peptidase 1, an aspartate protease belonging to the same family of proteases as the HIV protease. To search for low molecular weight peptide ligands,...
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Impact of selected emulsifiers and different emulsification methods on the growth of Staphylococcus aureus and Escherichia coli.
KOLDOVÁ, Kamila
The main topic of this bachelor's thesis is to determine the minimum inhibitory and bactericidal concentration of selected types of emulsifiers and plant extracts on the growth of Escherichia coli and Staphylococcus aureus bacteria. The thesis also examines the effect of selected emulsifiers and their methods of emulsification on emulsion stability. The theoretical part provides basic information about the bacteria used, their morphological characteristics, pathogenesis, and the diseases they cause. It also describes emulsifiers and their general characteristics, including emulsion stability. The final theoretical part of the thesis focuses on plant extracts, their properties, and their applications. The methodology section of this thesis illustrates the procedures for preparing emulsifiers and plant extracts, as well as determining the minimum inhibitory and bactericidal concentrations. These determinations were performed in a 96-well microtiter plate and measured spectrophotometrically. The method for preparing emulsion stability was conducted in glass test tubes, Eppendorf tubes, and centrifuge tubes. Based on these methodts, the stability of emulsion of Mueller-Hinton broth and sugar solution was determined. The results of minimum inhibitory and bactericidal concentration were processed into tables and evaluated. Emulsion stability was processed and evaluated using obtained photographs.
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Activated sludge inhibited by pesticides
Pokluda, Michal ; Pavlíková, Marcela (referee) ; Hrich, Karel (advisor)
The bachelor's thesis focuses on researching the inhibitory effects of selected pesticide substances on activated sludge present in wastewater treatment plants. The thesis is divided into a theoretical part and a practical part. The first part provides a theoretical explanation of pesticides, their presence in water, their basic classification, and the legislation addressing the issue of pesticide substances in water. The thesis also lists and describes methods for removing pesticides from water. Furthermore, the content of the theoretical part touches upon the topic of the Modřice wastewater treatment plant, primarily describing its treatment process. The practical part of the thesis involves laboratory research on the oxygen consumption rate of microorganisms in activated sludge in the presence of pesticides toxic to these organisms. The testing methodology is based on the standard ČSN EN ISO 8192.
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Interaction of the anticancer drug lenvatinib with cytochrome P450 subfamily 2C
Srogoň, Jiří ; Dračínská, Helena (advisor) ; Hýsková, Veronika (referee)
Lenvatinib is a multi-targeted kinase inhibitor capable of inhibiting these kinases at nanomolar concentrations. For this reason, it is used as a drug in the treatment of various types of cancer. Like many other xenobiotics, lenvatinib is metabolized by cytochromes P450, which can cause occurrence of drug interactions with other substances. Identification of the most important and clinically relevant drug interactions is essential to ensure the safety of patients already suffering from reduced quality of life due to cancer. The main aim of this bachelor thesis was to investigate the effect of lenvatinib on cytochromes P450, specifically on isoforms of subfamily 2C in humans and rats. The inhibitory effect was measured in vitro using marker rections on rat liver microsome samples for the CYP2C6 isoform and on human recombinant CYP2C9 isoform samples expressed in SuperosomesTM or Bactosomes® . Furthermore, the effect of lenvatinib on CYP2C11 and 2C6 expression in the liver of rats exposed to lenvatinib was investigated. Lenvatinib caused a decrease in the activity of the human recombinant CYP2C9 isoform, whereas no effect was observed on the activity of the CYP2C6 isoform in rat liver microsomes. One of the other objectives was to determine the IC50 for CYP2C9, but the objective was not met. When...
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Amaryllidaceae alkaloids as inspiration for preparation of selective butyrylcholinesterase inhibitors II
Králová, Kristýna ; Cahlíková, Lucie (advisor) ; Kučera, Tomáš (referee)
CHARLES UNIVERSITY FACULTY OF PHARMACY IN HRADEC KRÁLOVÉ DEPARTMENT OF PHARMACOGNOSY AND PHARMACEUTICAL BOTANY CANDIDATE: Kristýna Králová SUPERVISOR: prof. Ing. Lucie Cahlíková, Ph.D. CONSULTANT: Mgr. Filip Pidaný TITLE OF DIPLOMA THESIS: Amaryllidaceae alkaloids as inspiration of preparation of selective butyrylcholinesterase II Plants of the family Amaryllidaceae are an important source of biologically active substances, which are labeled as Amaryllidaceae alkaloids (AAs). These natural products show a wide range of biological characteristics, for which they are also intensively studied. Their significant biological effects include antiviral, antibacterial, antifungal, anti-inflammatory, antiparasitic, and cytotoxic activity. Their other important characteristic is the ability to inhibit both cholinesterases: acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). Inhibition of AChE is used in the therapy of Alzheimer's disease (AD). Currently, in the treatment of the symptoms of this disease, one AA, galanthamine, is used. Recently a new structural type of AAs, carltonins, have been isolated from the plant Narcissus pseudonarcissus cv. Carlton. These substances showed a significant inhibitory effect against BuChE and became the inspiration for the synthesis of selective BuChE inhibitors....
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Effect of the anticancer drug cabozantinib on cytochrome P450 activity
Slobodníková, Eva ; Dračínská, Helena (advisor) ; Václavíková, Radka (referee)
Cabozantinib is an anti-cancer drug used mainly for the treatment of thyroid and renal cell carcinoma. It is classified as a low molecular weight selective tyrosine kinase inhibitor. Tyrosine kinases play a key role in signal transduction and regulation of many cellular processes such as growth, differentiation, and proliferation. The changes in the tyrosine kinase pathways are associated with the formation and progression of tumors where their growth is uncontrolled. Tyrosine kinase inhibitors act on tyrosine kinase receptors, thereby preventing the spread of cancer cells and slowing down the progression of cancer. Because cabozantinib, like other clinically used drugs, is metabolized by cytochromes P450, adverse drug interactions may occur that result in altered pharmacokinetics of the administered drugs and a consequent decrease in the efficacy of these drugs. In this diploma thesis, the effect of cabozantinib on the activity of the main enzymes of phase I biotransformation of xenobiotics, cytochromes P450, was investigated in vitro. The effect on the activity of both rat and human cytochrome P450 isoforms involved in xenobiotic metabolism was studied. CYP isoforms were predominantly incubated with cabozantinib at two concentrations; 10 µM and a concentration corresponding to the substrate...
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