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Cytotoxicita beauvericinu, citrininu, deoxynivalenolu a T-2 toxinu stanovena metodou in vitro v buňkách Vero
Macáková, Petra ; Štaud, František (advisor) ; Vopršalová, Marie (referee)
Mycotoxins are secondary metabolites of moulds. Contamination of food and feed by mycotoxins is a major problem for human and animal health. Ingestion of mycotoxins may cause a range of toxic responses, from acute toxicity to long term or chronic health disorders. Several mycotoxins, either from the same or from different fungal species, occur simultaneously in plant products. However, its implication for food safety assessment is generally not known, as there is relatively little information on the interaction between concomitantly occurring mycotoxins and the consequence for the toxicity. Mycotoxins with similar mode of action would be expected to have at least additive effects. Conversely, some interactions could have subtractive effects. An understanding mode of action in simple in vitro systems can provide a rational basses for predicting interactions between mycotoxins. The aim of this study was to obtain cytotoxicity data (EC50 values) of Penicillium and Fusarium mycotoxins, nominally BEA, CIT, DON and T-2 toxin. For this purpose, Vero cells viability was evaluated in the presence of these four mycotoxins using the NR assay. All mycotoxins tested diminished cell viability in a concentration and incubation time-dependent manner on Vero cells. Individual mycotoxins increase cytotoxicity as...
Intracellular pH homeostasis in pathogenic yeast Candida albicans and Candida glabrata
Winterová, Lucie ; Štaud, František (advisor) ; Červený, Lukáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucie Winterová Supervisor: prof. PharmDr. František Štaud, Ph.D. Consultants: RNDr. Marie Kodedová, Ph.D., RNDr. Hana Sychrová, DrSc. Title of diploma thesis: Intracellular pH homeostasis in pathogenic yeast Candida albicans and Candida glabrata The thesis examines the influence of culture conditions on intracellular pH homeostasis in Candida albicans and Candida glabrata yeast strains with deletions of alkali- metal-cation membrane transporters, Cacnh1Δ and Cgtrk1Δ. Intracellular pH was measured with the use of pHluorin: a variant of a green fluorescent protein which had been expressed in the cytosol of both yeast species. Fluorescence of the expressed pHluorin was confirmed by a fluorescence microscopy and a calibration curve was created to determine the dependence of fluorescence intensity of pHluorin on intracellular pH. This thesis further demonstrates impact of medium composition (especially different nitrogen sources) and antifungal agents (fluconazole, clotrimazole, amphotericin B and terbinafine) on intracellular pH values in both yeasts. The effects of Cacnh1Δ and Cgtrk1Δ mutations were established on certain physiological parameters, such as the growth speed under different culture...
Study of interactions of antiviral drugs with selected placental transporters
Karbanová, Sára ; Štaud, František (advisor) ; Kacerovský, Marian (referee) ; Mladěnka, Přemysl (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Sára Karbanová, MSc. Supervisor: prof. PharmDr. František Štaud, Ph.D. Consultant: doc. PharmDr. Lukáš Červený, Ph.D. Title of Dissertation Thesis: Study of interactions of antiviral drugs with selected placental transporters The backbone of HIV treatment in pregnant women is the combination antiretroviral therapy which effectively suppresses the viral proliferation in maternal blood circulation. One of drugs in this therapy regimen should be a molecule with high placental transfer to assure the prophylaxis of viral transfer to fetus. There is no approved guideline for pharmacotherapy of pregnant women infected with hepatitis C (HCV), however recent evidence suggests that lowering the maternal viral load could correlate with lower likelihood of HCV transmission to the fetus. To assure and further develop the effective and safe pharmacotherapeutic approaches for treatment of HIV and HCV in pregnancy with respect to fetal safety, it is essential to understand mechanisms of placental transfer of anti(retro)viral drugs. In this Dissertation thesis we focused on molecules derived from nucleosides (anti-HIV abacavir, emtricitabine, zidovudine, tenofovir disoproxil fumarate and anti-HCV...
Study of interactions of antiviral drugs with selected placental transporters
Karbanová, Sára ; Štaud, František (advisor) ; Kacerovský, Marian (referee) ; Mladěnka, Přemysl (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Sára Karbanová, MSc. Supervisor: prof. PharmDr. František Štaud, Ph.D. Consultant: doc. PharmDr. Lukáš Červený, Ph.D. Title of Dissertation Thesis: Study of interactions of antiviral drugs with selected placental transporters The backbone of HIV treatment in pregnant women is the combination antiretroviral therapy which effectively suppresses the viral proliferation in maternal blood circulation. One of drugs in this therapy regimen should be a molecule with high placental transfer to assure the prophylaxis of viral transfer to fetus. There is no approved guideline for pharmacotherapy of pregnant women infected with hepatitis C (HCV), however recent evidence suggests that lowering the maternal viral load could correlate with lower likelihood of HCV transmission to the fetus. To assure and further develop the effective and safe pharmacotherapeutic approaches for treatment of HIV and HCV in pregnancy with respect to fetal safety, it is essential to understand mechanisms of placental transfer of anti(retro)viral drugs. In this Dissertation thesis we focused on molecules derived from nucleosides (anti-HIV abacavir, emtricitabine, zidovudine, tenofovir disoproxil fumarate and anti-HCV...
The influence of antidepressants on serotonin homeostasis in placenta
Váchalová, Veronika ; Štaud, František (advisor) ; Jirkovský, Eduard (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Veronika Váchalová Supervisor: Prof. PharmDr. František Štaud, Ph.D. Consultant: Mgr. Rona Karahoda Title of diploma thesis: The influence of antidepressants on serotonin homeostasis in placenta Depression is a serious mental disorder affecting 10-20% of women during pregnancy. Up to 10% of these pregnant women are prescribed antidepressants (ADs), most frequently from the class of selective serotonin (5-HT) reuptake inhibitors (SSRIs). While the safety of this treatment is questionable due to reported impaired pregnancy/fetal outcomes, understanding of potential mechanistic causes is still lacking. During pregnancy, 5-HT is important for normal placental function and proper fetal development and programming. 5-HT homeostasis in the placenta is maintained via the 5-HT transporter (SERT/SLC6A4) on the apical side and the recently characterized organic cation transporter 3 (OCT3/SLC22A3) on the basal side of trophoblast. These transporters take up 5-HT from the maternal and fetal circulations, respectively into the syncytiotrophoblast (STB) where it is degraded by monoamine oxidase-A (MAO-A). As all ADs interfere with the 5-HT system it is important to study their potential interactions in the...
Intracellular pH homeostasis in pathogenic yeast Candida albicans and Candida glabrata
Winterová, Lucie ; Štaud, František (advisor) ; Červený, Lukáš (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucie Winterová Supervisor: prof. PharmDr. František Štaud, Ph.D. Consultants: RNDr. Marie Kodedová, Ph.D., RNDr. Hana Sychrová, DrSc. Title of diploma thesis: Intracellular pH homeostasis in pathogenic yeast Candida albicans and Candida glabrata The thesis examines the influence of culture conditions on intracellular pH homeostasis in Candida albicans and Candida glabrata yeast strains with deletions of alkali- metal-cation membrane transporters, Cacnh1Δ and Cgtrk1Δ. Intracellular pH was measured with the use of pHluorin: a variant of a green fluorescent protein which had been expressed in the cytosol of both yeast species. Fluorescence of the expressed pHluorin was confirmed by a fluorescence microscopy and a calibration curve was created to determine the dependence of fluorescence intensity of pHluorin on intracellular pH. This thesis further demonstrates impact of medium composition (especially different nitrogen sources) and antifungal agents (fluconazole, clotrimazole, amphotericin B and terbinafine) on intracellular pH values in both yeasts. The effects of Cacnh1Δ and Cgtrk1Δ mutations were established on certain physiological parameters, such as the growth speed under different culture...
LC-MS analysis of phase one biotransformation of K1277, a potential drug against Alzheimer's disease
Krchová, Lucie ; Štaud, František (advisor) ; Kučera, Radim (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucie Krchová Supervisor: Prof. PharmDr. František Štaud, Ph.D. Supervisor - Consultant: Mgr. et Mgr. Rafael Doležal, Ph.D. Title of diploma thesis: LC-MS analysis of phase one biotransformation of K1277, a potential drug against Alzheimer's disease. Alzheimer's disease belongs to the most common neurodegenerative diseases that several milions of people all around the world suffer from. There are few symptomatic drugs, which are able to reduce signs and symptoms of this type of dementia, however none of them is able to completely stop it. The diploma thesis is focused on a metabolomic study of compound K1277, synthesized as a combination of 6-chlortacrine and aminoacid tryptophan, which thanks to its properties belongs to promising potential drugs against Alzheimer's disease and which has not been published yet. The aim of this diploma thesis was to perform in vitro metabolomic screening of the compound K1277 using human liver microsomes focusing on qualitative and quantitative analysis. As intermediate aims of the thesis, necessary conditions for sufficient biotransformation of compound K1277 were investigated, quantity of metabolised compound K1277, identifications of particular metabolites...
Interactions of antiretrovirals with drug transporters; role in pharmacokinetics
Řezníček, Josef ; Štaud, František (advisor) ; Chládek, Jaroslav (referee) ; Trejtnar, František (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Mgr. Josef Řezníček Supervisor: prof. PharmDr. František Štaud, Ph.D. Consultant: doc. PharmDr. Martina Čečková, Ph.D. Title of doctoral thesis: Interactions of antiretrovirals with drug transporters; role in pharmacokinetics Current pharmacotherapy of HIV positive patients consists of co-administration of three or more antiretrovirals from different pharmacotherapeutic groups, so called combination antiretroviral therapy (cART). Using this approach, a significant reduction in viral load, delayed viral resistance progression and prolonged efficacy of therapy is achieved. However, the use of cART often bears the risk of drug-drug interactions, which may result in subtherapeutic or supratherapeutic concentrations of drugs in organism with subsequent failure of therapy, or manifestation of toxic effects. Drug transporters expressed in many tissues of human body are widely responsible for occurrence of drug-drug interactions. Therefore, detailed knowledge on antiretrovirals pharmacokinetics and their interactions with drug transporters is important to ensure safe and effective therapy of HIV infection. The aim of this thesis was to study interactions of drugs used in combination antiretroviral...
LC-MS analysis of phase one biotransformation of K1277, a potential drug against Alzheimer's disease
Krchová, Lucie ; Štaud, František (advisor) ; Kučera, Radim (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lucie Krchová Supervisor: Prof. PharmDr. František Štaud, Ph.D. Supervisor - Consultant: Mgr. et Mgr. Rafael Doležal, Ph.D. Title of diploma thesis: LC-MS analysis of phase one biotransformation of K1277, a potential drug against Alzheimer's disease. Alzheimer's disease belongs to the most common neurodegenerative diseases that several milions of people all around the world suffer from. There are few symptomatic drugs, which are able to reduce signs and symptoms of this type of dementia, however none of them is able to completely stop it. The diploma thesis is focused on a metabolomic study of compound K1277, synthesized as a combination of 6-chlortacrine and aminoacid tryptophan, which thanks to its properties belongs to promising potential drugs against Alzheimer's disease and which has not been published yet. The aim of this diploma thesis was to perform in vitro metabolomic screening of the compound K1277 using human liver microsomes focusing on qualitative and quantitative analysis. As intermediate aims of the thesis, necessary conditions for sufficient biotransformation of compound K1277 were investigated, quantity of metabolised compound K1277, identifications of particular metabolites...
Interactions of antiretrovirals with drug transporters; role in pharmacokinetics
Řezníček, Josef ; Štaud, František (advisor) ; Chládek, Jaroslav (referee) ; Trejtnar, František (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Mgr. Josef Řezníček Supervisor: prof. PharmDr. František Štaud, Ph.D. Consultant: doc. PharmDr. Martina Čečková, Ph.D. Title of doctoral thesis: Interactions of antiretrovirals with drug transporters; role in pharmacokinetics Current pharmacotherapy of HIV positive patients consists of co-administration of three or more antiretrovirals from different pharmacotherapeutic groups, so called combination antiretroviral therapy (cART). Using this approach, a significant reduction in viral load, delayed viral resistance progression and prolonged efficacy of therapy is achieved. However, the use of cART often bears the risk of drug-drug interactions, which may result in subtherapeutic or supratherapeutic concentrations of drugs in organism with subsequent failure of therapy, or manifestation of toxic effects. Drug transporters expressed in many tissues of human body are widely responsible for occurrence of drug-drug interactions. Therefore, detailed knowledge on antiretrovirals pharmacokinetics and their interactions with drug transporters is important to ensure safe and effective therapy of HIV infection. The aim of this thesis was to study interactions of drugs used in combination antiretroviral...

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