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Frequency of occurrence of selected single nucleotide polymorphisms of CYP2C8 and MDR1 in the Czech population and their influence on the effect of amiodarone
Pechandová, Kristina ; Perlík, František (advisor) ; Král, Jiří (referee) ; Anzenbacher, Pavel (referee)
Frekvence výskytu vybraných bodových polymorfismů CYP2C8 a MDR1 v české populaci a jejich vliv na působení amiodaronu Úvod: Variabilita lékové odpovědi je někdy podmíněna genetickými rozdíly v metabolismu a transportu léčiv. Interindividuální rozdíly jsou často způsobeny polymorfismy, které ovlivňují biotransformační aktivitu enzymů a expresi transportérů. V disertační práci jsme věnovali pozornost cytochromu P450 izoenzymu CYP2C8 a MDR1. Nejprve jsme popsali frekvenci výskytu vybraných variantních alel CYP2C8*2, CYP2C8*3 (2 substituce v exonu 3 a 8, CYP2C8*3G416A a CYP2C8*3A1196G), CYP2C8*4, CYP2C8 P404A u zdravé české populace a variantních alel MDR1 v exonech: 26 C3435T, 21 G2677A/T, 12 C1236T a 17 T-76A. Následně jsme sledovali vliv těchto polymorfismů na působení amiodaronu u vybraného souboru pacientů. Metody: Genotyp MDR1 a CYP2C8 jsme stanovili pomocí PCR-RFLP za využití specifických restrikčních enzymů a primerů. Frekvence genotypů MDR1jsme určili u 189 zdravých dobrovolníků a CYP2C8 u 161 zdravých osob. Do sledování jsme dále zařadili 63 pacientů užívajících amiodaron déle než dva měsíce. Jejich léčbu jsme posuzovali ze záznamů lékařské dokumentace, s využitím standardních biochemických a hematologických vyšetření a záznamů EKG. Koncentrace amiodaronu a jeho metabolitu N-...
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Interaction of Organophosphorus Poisoning Antidotes with Muscarinic and Nitotinic Receptors
Soukup, Ondřej ; Fusek, Josef (advisor) ; Trejtnar, František (referee) ; Anzenbacher, Pavel (referee)
1 Summary in English Currently, organophosphorus (OP) poisoning is a threat particularly in war conflicts in connection with terrorism or a poisoning caused by OP pesticides in agriculture. The poisoning is caused by inhibition of the enzyme acetylcholinesterase (AChE), which, under physiological conditions, cleaves acetylcholine (ACh) and regulates the transmission of nerve signals. The inhibition of AChE leads to excessive stimulation of cholinergic receptors and clinically it manifests as so-called cholinergic syndrome. According to the structures affected, symptoms are divided into nicotinic, muscarinic and central. Organophosphates are lethal compounds, when death is caused by suffocation. Current treatment is mainly based on the application of anticholinergics (atropine) and /or on the application of AChE oxime reactivators (HI-6, obidoxime, pralidoxime etc.) Atropine inhibits excessive neural transmission by blocking muscarinic receptors. However, it is only symptomatic cure not causal. Whereas oximes, which represent a causal therapy, are able to unbind an OP compound from the AChE and restore its splitting function. Using reactivators has two major drawbacks; there is no universal reactivator against all types of OP compounds (soman, sarin, paraoxon etc.) and reactivation is possible only within a...
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Frequency of occurrence of selected single nucleotide polymorphisms of CYP2C8 and MDR1 in the Czech population and their influence on the effect of amiodarone
Pechandová, Kristina ; Perlík, František (advisor) ; Král, Jiří (referee) ; Anzenbacher, Pavel (referee)
Frequency of occurrence of selected single nucleotide polymorphisms of CYP2C8 and MDR1 in the Czech population and their influence on the effect of amiodarone Introduction: Variability in drug response is sometimes conditioned by genetic differences in the metabolism and the transport of drugs. Interindividual differences are often caused by polymorphisms affecting biotransformation activity of enzymes and expression of transporters. In the thesis we paid attention to the cytochrome P450 CYP2C8 and MDR1. First, we described the frequency of occurrence of selected variant alleles CYP2C8 * 2, CYP2C8 * 3 (2 substitution in exon 3 and 8, CYP2C8 and CYP2C8 * 3G416A * 3A1196G), CYP2C8 * 4, CYP2C8 P404A in the healthy Czech population and MDR1 variant alleles in these exons: 26 C3435T, 21 G2677A/T, 12 C1236T a 17 T-76A. Subsequently, we studied the influence of these polymorphisms on effects of amiodarone in the selected group of patients. Methods: We determined genotypes MDR1 a CYP2C8 by PCR-RFLP by using restriction enzymes and specific primers. We determined the frequency of MDR1 genotypes in 189 healthy volunteers and CYP2C8 in 161 healthy subjects. Further we included into the study 63 patients treated with amiodarone for longer than two months. Their treatment was assessed from medical records and...
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Frequency of occurrence of selected single nucleotide polymorphisms of CYP2C8 and MDR1 in the Czech population and their influence on the effect of amiodarone
Pechandová, Kristina ; Perlík, František (advisor) ; Král, Jiří (referee) ; Anzenbacher, Pavel (referee)
Frekvence výskytu vybraných bodových polymorfismů CYP2C8 a MDR1 v české populaci a jejich vliv na působení amiodaronu Úvod: Variabilita lékové odpovědi je někdy podmíněna genetickými rozdíly v metabolismu a transportu léčiv. Interindividuální rozdíly jsou často způsobeny polymorfismy, které ovlivňují biotransformační aktivitu enzymů a expresi transportérů. V disertační práci jsme věnovali pozornost cytochromu P450 izoenzymu CYP2C8 a MDR1. Nejprve jsme popsali frekvenci výskytu vybraných variantních alel CYP2C8*2, CYP2C8*3 (2 substituce v exonu 3 a 8, CYP2C8*3G416A a CYP2C8*3A1196G), CYP2C8*4, CYP2C8 P404A u zdravé české populace a variantních alel MDR1 v exonech: 26 C3435T, 21 G2677A/T, 12 C1236T a 17 T-76A. Následně jsme sledovali vliv těchto polymorfismů na působení amiodaronu u vybraného souboru pacientů. Metody: Genotyp MDR1 a CYP2C8 jsme stanovili pomocí PCR-RFLP za využití specifických restrikčních enzymů a primerů. Frekvence genotypů MDR1jsme určili u 189 zdravých dobrovolníků a CYP2C8 u 161 zdravých osob. Do sledování jsme dále zařadili 63 pacientů užívajících amiodaron déle než dva měsíce. Jejich léčbu jsme posuzovali ze záznamů lékařské dokumentace, s využitím standardních biochemických a hematologických vyšetření a záznamů EKG. Koncentrace amiodaronu a jeho metabolitu N-...
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Interaction of Organophosphorus Poisoning Antidotes with Muscarinic and Nitotinic Receptors
Soukup, Ondřej ; Fusek, Josef (advisor) ; Trejtnar, František (referee) ; Anzenbacher, Pavel (referee)
1 Summary in English Currently, organophosphorus (OP) poisoning is a threat particularly in war conflicts in connection with terrorism or a poisoning caused by OP pesticides in agriculture. The poisoning is caused by inhibition of the enzyme acetylcholinesterase (AChE), which, under physiological conditions, cleaves acetylcholine (ACh) and regulates the transmission of nerve signals. The inhibition of AChE leads to excessive stimulation of cholinergic receptors and clinically it manifests as so-called cholinergic syndrome. According to the structures affected, symptoms are divided into nicotinic, muscarinic and central. Organophosphates are lethal compounds, when death is caused by suffocation. Current treatment is mainly based on the application of anticholinergics (atropine) and /or on the application of AChE oxime reactivators (HI-6, obidoxime, pralidoxime etc.) Atropine inhibits excessive neural transmission by blocking muscarinic receptors. However, it is only symptomatic cure not causal. Whereas oximes, which represent a causal therapy, are able to unbind an OP compound from the AChE and restore its splitting function. Using reactivators has two major drawbacks; there is no universal reactivator against all types of OP compounds (soman, sarin, paraoxon etc.) and reactivation is possible only within a...
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Panels of steroid receptor reporter cell lines for compound profiling and development of selective ligands for estrogen receptor alpha and beta
Sedlák, David ; Bartůněk, Petr (advisor) ; Anzenbacher, Pavel (referee) ; Kasal, Alexander (referee)
of the Ph.D. thesis Steroid hormone receptors represent a major target in the drug discovery. As ligand inducible transcription factors, their activity can be modulated by small lipophilic molecules. The first part of this work describes a preparation of two panels of potent, selective and robust luciferase reporter cell lines on the unified cellular background in U2OS osteosarcoma cell line. This system consists of two panels of stable luciferase reporter cell lines for estrogen receptor α (ERα), estrogen receptor β (ERβ), androgen receptor (AR), glucocorticoid receptor (GR), mineralocorticoid receptor (MR) and progesterone receptor (PR). The first panel of reporter cell lines relies on the expression of the chimeric steroid receptors created by the replacement of the N-terminal part of the steroid receptor molecule by Gal4 DNA binding domain (Gal4 DBD) binding to 9 copies of Gal4 upstream activation sequences (Gal4 UAS) in the promoter of the pGL4 luciferase reporter vector. In the second panel of reporter cell lines the activation of either synthetic promoter containing multiple hormone response elements or viral promoter derived from MMTV LTR is mediated by full- length exogenously expressed steroid receptors. We have extensively validated both panels using 28 well established ligands, carefully...
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Importance of Transporter Proteins for Fetal Protection
Hahnová, Lenka ; Štaud, František (advisor) ; Anzenbacher, Pavel (referee) ; Geršl, Vladimír (referee)
Doctoral thesis Mgr. Lenka Hahnová (Cygalová) Placenta, which represents a unique link between the mother and fetus, fulfills many functions essential for normal course of pregnancy and uncomplicated development of the fetus. Nutrient supply, gas exchange and metabolic waste product removal belong to its main roles. In addition, placenta serves as an endocrine, metabolic, immune and protective organ, since during pregnancy mother may be, either unconsciously or deliberately, exposed to a wide range of substances toxic for the fetus. Originally, it was supposed that the physiological barrier between maternal and fetal circulation is created only by cellular layers of syncytiotrophoblast and endothelium of fetal capillaries. However, it has been demonstrated that besides this mechanical component of protection, activity of drug efflux transporters and metabolic enzymes localized in the polarized syncytiotrophoblast layer contribute considerably to the protective function of placental barrier. Efflux transporters are ATP dependent membrane proteins capable of actively removing different molecules out of cells. So far, the best described drug efflux transporters are P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP), significantly affecting kinetics of transplacental passage of various...
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