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Interaction of selected anti-HCV drugs with placental OCTN2 transport protein
Machalová, Vanda ; Čečková, Martina (advisor) ; Hyršová, Lucie (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Mgr. Vanda Machalová Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Title of rigorous thesis: Interaction of selected anti-HCV drugs with placental OCTN2 transport protein. The aim of this rigorous work was to test the interaction of paritaprevir and daclatasvir with the placental OCTN2 transporter, which ensures the transport of L-carnitine as a cofactor of the fatty acid oxidation process to obtain sufficient energy for correct intra-arterial fusion growth. No drugs in this group have been approved for the treatment of chronic hepatitis C in pregnancy, therefore examining their effect on the transport of L-carnitine via OCTN2 can create certain image concerning to the fetal and maternal safety. In the first part of my thesis the transport and accumulation of radioactively labeled L- carnitine via monolayers of BeWo b30 cells in the presence of paritaprevir and daclatasvir was assayed and the concentration of radiolabeled L-carnitine inside the cell layer. In the second part of my thesis we focused on the evaluation of the presence of paritaprevir and daclatasvir on its gene expression of the OCTN1 and OCTN2 carnitine transporters (or their coding genes SLC22A4 and SLC22A5) by qRT-PCR. The...
Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study
Zubatá, Karolína ; Čečková, Martina (advisor) ; Hronek, Miloslav (referee)
Charles University University of Porto Faculty of Pharmacy in Hradec Králové Faculty of Pharmacy Department of Pharmacology and Toxicology Department of Biological Sciences Student: Karolína Zubatá Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Consultants: Susana Rocha, Ph.D., prof. Alice Santos-Silva, Ph.D. Title of diploma thesis: Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study Increased levels of oxidative stress (OS) have been described in healthy, full-term newborns as a consequence of the drastic changes introduced by birth and by the exposure to extrauterine environment. Our intention was to examine the OS levels in red blood cells (RBCs) of neonates and to further understand the changes that the newborn organism undergoes with its newly- acquired autonomy as this knowledge is limited and there are no reference values. Umbilical cord blood samples were collected from a small population of newborns (n = 8) and several hematological and biochemistry parameters were evaluated. Our experimental data consist of OS biomarkers measurements performed in different fractions of blood (RBC membrane, total RBCs and plasma): membrane bound hemoglobin (MBH), lipid peroxidation (LPO), quantification of catalase (CAT) and glutathione peroxidase (GPx) activities,...
Interactions of selected anticancer drugs of the MAPK/ERK signaling pathway inhibitors group with the ABC drug transporters
Slatinský, Lukáš ; Čečková, Martina (advisor) ; Hofman, Jakub (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lukáš Slatinský Supervisor: Assoc. prof. PharmDr. Martina Čečková, Ph.D. Title of diploma thesis: Interactions of selected anticancer drugs of the MAPK/ERK signaling pathway inhibitors group with the ABC drug transporters ABCB1 (Pgp, P-glycoprotein) and ABCG2 (BCRP, breast cancer resistance protein) are members of a transmembrane efflux ATP dependent transporter family, so called ATP-binding cassettes (ABC). Physiologicaly they are expressed in the cellular membrane and protect body tissues against potentially toxic xenobiotics including drugs. They represent also one of the tumor defense mechanisms when being able to efflux a wide variety of cytotoxic drugs out of the cancer cells leading to treatment failure. BRAF protein plays an important regulatory and signal role in MAPK/ERK pathway affecting cell division, differentiation and secretion. Mutations of BRAF lead to overactivity in MAPK/ERK pathway in many cancer cells and can be therefore targeted by anticancer therapy. Cobimetinib and dabrafenib are relatively new anticancer therapeutics inhibiting the signal pathway mentioned above and they are used in treatment of melanoma carrying the BRAF mutation. The aims of this project were to...
In vitro transport of abacavir across the monolayer of Caco-2 cells; interaction with etravirine and rilpivirine
Mlčochová, Alice ; Čečková, Martina (advisor) ; Červený, Lukáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Alice Mlčochová Supervisor: PharmDr. Martina Čečková, Ph.D. Title of diploma thesis: In vitro transport of abacavir across the monolayer of Caco-2 cells; interaction with etravirine and rilpivirine. Abacavir belongs among nucleoside reverse transciptase inhibitors (NRTIs) representing a basic component of combined antiretroviral therapy used in treatment of HIV-positive patients [1]. Etravirine and rilpivirine are newer non-nucleoside reverse transcriptase inhibitors (NNRTIs) combined in cART together with NRTI. ATP-dependent transporters, so called ABC transporters, are able to affect pharmacokinetic properties of drugs, thus they are important site of drug-drug interactions affecting absorption, distribution and excretion level. P-glycoprotein (Pgp, ABCB1) and BCRP (ABCG2) belong among the most clinically important ABC transporters able to cause drug-drug interactions. The aim of this thesis was to introduce and optimize the method for evaluation of drug absorption using monolayers of Caco-2human intestine cell lines, whose integrity was verified by evaluating TEER (transepithelial electrical resistance). This model was also used for abacavir transport studies. Significant...
Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study
Zubatá, Karolína ; Čečková, Martina (advisor) ; Hronek, Miloslav (referee)
Charles University University of Porto Faculty of Pharmacy in Hradec Králové Faculty of Pharmacy Department of Pharmacology and Toxicology Department of Biological Sciences Student: Karolína Zubatá Supervisor: doc. PharmDr. Martina Čečková, Ph.D. Consultants: Susana Rocha, Ph.D., prof. Alice Santos-Silva, Ph.D. Title of diploma thesis: Oxidative stress biomarkers of the erythrocyte in the newborn - a follow-up study Increased levels of oxidative stress (OS) have been described in healthy, full-term newborns as a consequence of the drastic changes introduced by birth and by the exposure to extrauterine environment. Our intention was to examine the OS levels in red blood cells (RBCs) of neonates and to further understand the changes that the newborn organism undergoes with its newly- acquired autonomy as this knowledge is limited and there are no reference values. Umbilical cord blood samples were collected from a small population of newborns (n = 8) and several hematological and biochemistry parameters were evaluated. Our experimental data consist of OS biomarkers measurements performed in different fractions of blood (RBC membrane, total RBCs and plasma): membrane bound hemoglobin (MBH), lipid peroxidation (LPO), quantification of catalase (CAT) and glutathione peroxidase (GPx) activities,...
The effect of lipid signaling pathway interference on sorafenib cytotoxic efficacy and function of efflux transporters in mouse hepatocellular carcinoma cells
Sagandykova, Aigul ; Čečková, Martina (advisor) ; Novotná, Eva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Aigul Sagandykova Supervisor: PharmDr. Martina Čečková, Ph.D. Consultant: Dr. Mikko Gynther Ph.D. Title of diploma thesis: The effect of lipid signalling pathway interference on sorafenib cytotoxic efficacy and function of efflux transporters in mouse hepatocellular carcinoma cells. Nowadays cancer remains one of the most challenging health issues worldwide. Chemotherapy represents one of the essential approaches in the treatment of malignant diseases. However, multidrug resistance (MDR), a multifactorial phenomenon described as a loss of sensitivity of cancer cells to several diverse chemotherapeutic agents at the same time, often compromises the therapy outcomes. A well-known cause of MDR is an increased expression or/and an enhanced activity of efflux drug transporters of ATP binding cassette (ABC) superfamily, which has been found in many types of cancer. In the last decade, an expanding body of literature suggested a new hallmark of cancer cells - inflammation. An inflammatory microenvironment potentiates tumorigenesis and upregulation of transporters. Moreover, several observations show that ABC transporters mediate the transport of some signalling lipids. This new insight provided...
The role of biotransformation enzymes in the resistance of cancer cells against standard cytostatics
Giannitsi, Anna ; Hofman, Jakub (advisor) ; Čečková, Martina (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Anna Giannitsi Supervisor: RNDr. Jakub Hofman, Ph.D Title of diploma thesis: The role of biotransformation enzymes in the resistance of cancer cells against standard cytostatics Drug resistance is currently one of the major problems of chemotherapy. Tumor cells are able to defend themselves against the effect of cytostatic drugs due to various mechanisms which leads to a failure of anticancer therapy. The effort to describe new mechanisms of resistance and to develop new therapeutic methods, which would limit this therapeutic obstacle, is logically the subject of many studies. The activity of drug metabolizing enzymes and the subsequent decrease of intercellular concentration of anticancer drugs belongs to one of the possible mechanisms of pharmacokinetic resistance. Enzymes of I. and II. phase of biotransformation participate in this phenomenon. Cytochromes P450, main enzymes of the I. phase, play a major role in the metabolism of many cytostatic agents producing either pharmacologically active or inactive metabolites. Increased expression in tumors and the involvement of individual isoforms into the overall metabolism of cytostatic, which is deactivated by their activity, seems to be one of the...
Interactions of selected anticancer drugs of the MAPK/ERK signaling pathway inhibitors group with the ABC drug transporters
Slatinský, Lukáš ; Čečková, Martina (advisor) ; Hofman, Jakub (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Lukáš Slatinský Supervisor: Assoc. prof. PharmDr. Martina Čečková, Ph.D. Title of diploma thesis: Interactions of selected anticancer drugs of the MAPK/ERK signaling pathway inhibitors group with the ABC drug transporters ABCB1 (Pgp, P-glycoprotein) and ABCG2 (BCRP, breast cancer resistance protein) are members of a transmembrane efflux ATP dependent transporter family, so called ATP-binding cassettes (ABC). Physiologicaly they are expressed in the cellular membrane and protect body tissues against potentially toxic xenobiotics including drugs. They represent also one of the tumor defense mechanisms when being able to efflux a wide variety of cytotoxic drugs out of the cancer cells leading to treatment failure. BRAF protein plays an important regulatory and signal role in MAPK/ERK pathway affecting cell division, differentiation and secretion. Mutations of BRAF lead to overactivity in MAPK/ERK pathway in many cancer cells and can be therefore targeted by anticancer therapy. Cobimetinib and dabrafenib are relatively new anticancer therapeutics inhibiting the signal pathway mentioned above and they are used in treatment of melanoma carrying the BRAF mutation. The aims of this project were to...
Pharmacotherapy of breast cancer: Novel approaches, overcoming of multidrug resistance
Menelaou, Pavlina ; Čečková, Martina (advisor) ; Pávek, Petr (referee)
Pavlina Menelaou: Pharmacotherapy of breast cancer: Novel approaches, overcoming of multidrug resistance. (Diploma thesis) ABSTRACT Breast cancer arises when there is an uncontrolled growth of abnormal cells in the breast. Most often the tumor involves glandular breast cells in the ducts or lobules. The most common sign of breast cancer is a new lump or mass. Other possible signs can be redness, swelling, nipple discharge and more. Some risk factors of breast cancer can be alcohol, obesity, oral hormonal contraceptives, radiation, gender, age, hereditary/genetic aspects, estrogen and progesterone receptors. Breast cancer can be treated by surgery including mastectomy, lymphedema surgery and lumpectomy, by radiation therapy or chemotherapy. The novel methods include hormonal therapy using selective estrogen receptor modulators like tamoxifen, raloxifene, aromatase inhibitors like anastrozole and letrozole. One of the promising therapies is immune therapy with the use of interferons and other cytokines, dendritic cells and vaccines. Another novel approach is represented by the targeted therapy that is using monoclonal antibodies, tyrosine kinase inhibitors and more promising ways. The gene therapy that is made to correct specific molecular defects that contributes to the cause or progressions of breast cancer...
Effect of epigallocatechin gallate on bile production
Hiršová, Petra ; Čečková, Martina (advisor) ; Trejtnar, František (referee) ; Slanař, Ondřej (referee)
Effect of epigallocatechin gallate Epigallocatechin gallate (EGCG), the major green tea catechin, has been shown to be protective in various experimental models of liver injury. Since its effect on biliary physiology and liver cholesterol homeostasis has not been thoroughly studied, the present study investigated effect of EGCG on bile flow, bile acid homeostasis and cholesterol metabolism in healthy and ethinylestradiol-treated rats. Compared to controls, EGCG treatment in rats decreased bile flow by 23%. Hepatic paracellular permeability and biliary bile acid excretion were not altered by EGCG administration, but biliary glutathione excretion was reduced by 70%. Accordingly, the main glutathione transporter at the hepatocyte canalicular membrane, multidrug resistance-associated protein 2 (Mrp2), was significantly decreased at the protein level. Interestingly, EGCG markedly enhanced biliary excretion of cholesterol and phospholipids. These changes tightly correlated with increased expression of ATP- binding cassette transporter G5 and G8 (Abcg5/8) and scavenger receptor class B type 1 and with decreased expression of acyl-CoA:cholesterol acyltransferase (Acat2). EGCG administration to rats also doubled plasma bile acid concentrations compared to controls. While protein expression of the main...

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1 ČEČKOVÁ, Monika
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