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Monitoring the antioxidant activity of newly synthesized potential acetylcholinesterase modulators
Keratová, Žaneta ; Macháček, Miloslav (advisor) ; Kobrlová, Tereza (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Žaneta Keratová Supervisor: RNDr. Miloslav Macháček, Ph.D. Consultant: RNDr. Ľubica Múčková, Ph.D. Title of diploma thesis: Monitoring the antioxidant activity of newly synthesized potential acetylcholinesterase modulators. The theoretical part of the diploma thesis focuses on the characteristics of oxidative stress (OS) and antioxidant protection of the organism. In the next part, neurodegenerative diseases and the importance of OS in their development are discussed in detail. In the experimental part, the cytotoxicity of new potential acetylcholinesterase modulators was evaluated using the neuroblastoma cell line SH-SY5Y. The cytotoxicity of the compounds, expressed as IC50, was measured by a colorimetric method using the tetrazolium salt (MTT). This method was also used to determine the maximum tolerated concentration (MTC). These values were subsequently validated by measuring the apoptotic profile of cells in a physiological environment (without OS) and hydrogen peroxide (H2O2)-induced OS using microcapillary flow cytometry. The results showed that the compounds did not induce any statistically significant change in the distribution of cells in each population. Subsequently, the antioxidant...
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Monitoring the antioxidant activity of newly synthesized potential acetylcholinesterase modulators
Keratová, Žaneta ; Macháček, Miloslav (advisor) ; Kobrlová, Tereza (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Žaneta Keratová Supervisor: RNDr. Miloslav Macháček, Ph.D. Consultant: RNDr. Ľubica Múčková, Ph.D. Title of diploma thesis: Monitoring the antioxidant activity of newly synthesized potential acetylcholinesterase modulators. The theoretical part of the diploma thesis focuses on the characteristics of oxidative stress (OS) and antioxidant protection of the organism. In the next part, neurodegenerative diseases and the importance of OS in their development are discussed in detail. In the experimental part, the cytotoxicity of new potential acetylcholinesterase modulators was evaluated using the neuroblastoma cell line SH-SY5Y. The cytotoxicity of the compounds, expressed as IC50, was measured by a colorimetric method using the tetrazolium salt (MTT). This method was also used to determine the maximum tolerated concentration (MTC). These values were subsequently validated by measuring the apoptotic profile of cells in a physiological environment (without OS) and hydrogen peroxide (H2O2)-induced OS using microcapillary flow cytometry. The results showed that the compounds did not induce any statistically significant change in the distribution of cells in each population. Subsequently, the antioxidant...
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Modulation of anti-inflammatory response in microglial cell line SIM-A9
Hapalová, Jitka ; Macháček, Miloslav (advisor) ; Kobrlová, Tereza (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Jitka Tumpachová Supervisor: RNDr. Miloslav Macháček, Ph.D. Consultant: pplk. doc. MUDr. Jaroslav Pejchal, Ph.D. et Ph.D. Title of diploma thesis: Modulation of anti-inflammatory response in microglial cell line SIM-A9 In the theoretical section, the thesis is focused on the characteristics, significance, and contribution of neuroinflammation in the origin and development of neurodegenerative diseases. The possibilities of their current treatment are discussed. The experimental part is devoted to evaluating the cytotoxicity and anti-inflammatory response of newly synthesized derivatives of naphthoquinones (K1502, K1503 and K1504) and chromones (5-EM-13, 5-EM-26 and 5-EM-36). Cytotoxicity was measured in the mouse microglial line SIM-A9. In addition, the neuroblastoma line SH-SY5Y was used comparatively in this part. The half-maximal inhibitory concentration (IC50) and maximum tolerated concentration (MTC) were determined on both cell lines using MTT colorimetric methods. The safety of MTC was subsequently verified by microcapillary flow cytometry in the SIM-A9 cells after lipopolysaccharide (LPS) or interferon-γ (IFN-γ) stimulation. The anti-inflammatory effects of the studied substances were...
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Analysis of Psilocybe constituents I.
Kobrlová, Tereza ; Jahodář, Luděk (advisor) ; Jáč, Pavel (referee)
The goal of this work was to determine the contents of two main alkaloid types of Psilocybe species occurring in the Czech Republic. Correct identification of species was checked by mycology specialist. Qualitative and quantitative determination of substances contained in three collected species of the genus Psilocybe was carried out by LC-MSn (LIT). Samples were mainly collected in the north and northeast Bohemia. Three species of Psilocybe were found. The most found species were P. serbica var. bohemica then P. semilanceata and only one deposit of P. serbica var. arcana. Total of 35 deposits within eighteen locations. From qualitative point of view, the two main alkaloids were confirmed in all samples. Quantitative results for content of psilocin and psilocybin (in this order) are: P. bohemica 0,005 - 0,152 %, 0,000 - 0,048 %; P. semilanceata 0,087 - 0,337 %, 0,000 - 0,005 %; P. arcana 0,018 - 0,031 %, 0,003 - 0,022 %. Results in all locations are similar and rather dependent on the fungi type. Our results are based on dry mass. This work brought fundamental knowledge for future optimisation of analytical methods and assumptions for the project extension of other locations and determination of content of minority substances alongside the content of majority substances.
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Prediction of the penetration of new drugs through the blood-brain barrier
Kobrlová, Tereza ; Soukup, Ondřej (advisor) ; Čečková, Martina (referee) ; Fusek, Josef (referee)
and keywords The dissertation thesis deals with problem of insufficient penetration of antidotes for treatment of organophosphorus poisonings. Methods for evaluating the ability of compounds to penetrate the central nervous system (CNS) were developed and compared to each other. These methods were subsequently used for the standard and newly synthesized potential drugs from this group to evaluate their penetration to the brain. Furthermore, the strategies that could improve CNS penetration were investigated. Highly toxic organophosphorus compounds represent a big threat due to possible misuse in the military of for terrorist purposes. These compounds affecting cholinergic neurotransmission by inhibition of the enzyme acetylcholinesterase (AChE). For this reason, they are called nerve agents (NAs). NAs are extremely toxic, relatively easily obtainable and the therapy of intoxication is insufficiently effective. One of the major obstacles of AChE reactivators, causal antidote used in the treatment, is to overcome the blood-brain barrier in therapeutic concentration and restore the function of AChE in the CNS. Thus, research and development of new drug candidates for such antidotes requires appropriate methods for evaluation of their biological properties even in the in vitro stage. Selection,...
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Modulation of anti-inflammatory response in microglial cell line SIM-A9
Tumpachová, Jitka ; Macháček, Miloslav (advisor) ; Kobrlová, Tereza (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Jitka Tumpachová Supervisor: RNDr. Miloslav Macháček, Ph.D. Consultant: pplk. doc. MUDr. Jaroslav Pejchal, Ph.D. et Ph.D. Title of diploma thesis: Modulation of anti-inflammatory response in microglial cell line SIM-A9 In the theoretical section, the thesis is focused on the characteristics, significance, and contribution of neuroinflammation in the origin and development of neurodegenerative diseases. The possibilities of their current treatment are discussed. The experimental part is devoted to evaluating the cytotoxicity and anti-inflammatory response of newly synthesized derivatives of naphthoquinones (K1502, K1503 and K1504) and chromones (5-EM-13, 5-EM-26 and 5-EM-36). Cytotoxicity was measured in the mouse microglial line SIM-A9. In addition, the neuroblastoma line SH-SY5Y was used comparatively in this part. The half-maximal inhibitory concentration (IC50) and maximum tolerated concentration (MTC) were determined on both cell lines using MTT colorimetric methods. The safety of MTC was subsequently verified by microcapillary flow cytometry in the SIM-A9 cells after lipopolysaccharide (LPS) or interferon-γ (IFN-γ) stimulation. The anti-inflammatory effects of the studied substances were...
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Analysis of Psilocybe constituents I.
Kobrlová, Tereza ; Jahodář, Luděk (advisor) ; Jáč, Pavel (referee)
The goal of this work was to determine the contents of two main alkaloid types of Psilocybe species occurring in the Czech Republic. Correct identification of species was checked by mycology specialist. Qualitative and quantitative determination of substances contained in three collected species of the genus Psilocybe was carried out by LC-MSn (LIT). Samples were mainly collected in the north and northeast Bohemia. Three species of Psilocybe were found. The most found species were P. serbica var. bohemica then P. semilanceata and only one deposit of P. serbica var. arcana. Total of 35 deposits within eighteen locations. From qualitative point of view, the two main alkaloids were confirmed in all samples. Quantitative results for content of psilocin and psilocybin (in this order) are: P. bohemica 0,005 - 0,152 %, 0,000 - 0,048 %; P. semilanceata 0,087 - 0,337 %, 0,000 - 0,005 %; P. arcana 0,018 - 0,031 %, 0,003 - 0,022 %. Results in all locations are similar and rather dependent on the fungi type. Our results are based on dry mass. This work brought fundamental knowledge for future optimisation of analytical methods and assumptions for the project extension of other locations and determination of content of minority substances alongside the content of majority substances.
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