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Biotransformation of flubendazole and albendazole in plant cells
Seidlová, Helena ; Skálová, Lenka (advisor) ; Karlíčková, Jana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Helena Seidlová Supervisor: Prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Biotransformation of flubendazole and albendazole in plant cells Benzimidazole anthelmintics, the drugs against parasitic worms, are widely used in human as well as in veterinary medicine. Following excretion, these substances may persist in the environment and impact non-target organisms. Phytoremediation might be a suitable method for elimination of anthelmintics from the environment. In order to test detoxification abilities of plants, biotransformation pathways of albendazole (ABZ) and flubendazole (FLU) were studied in reed (Phragmites australis) in vitro. It was found that reed cells were able to uptake and biotransform both anthelmintics. These drugs did not significantly affect viability of the reed cells. Using HPLC/MS 10 ABZ metabolites and 5 FLU metabolites were found. Some atypical biotransformation metabolites (glucosylglucosides, acetylglucosides and xylosylglucosides), which have not been previously reported were identified. Based on the obtained results, the schemes of metabolic pathways of ABZ and FLU in reed were proposed. The amount of main metabolites (reduced FLU, ABZ-sulfoxide,...
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Effect of inositol hexaphosphate on adhesion and migration of cell lines SW480 and SW620
Peterková, Alena ; Szotáková, Barbora (advisor) ; Skálová, Lenka (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Alena Ježková Supervisor: Doc. Ing. Barbora Szotáková, Ph.D. Supervisor specialist: RNDr. Veronika Hanušová, Ph.D. Title of diploma thesis: Effect of inositol hexaphosphate on adhesion and migration of cell lines SW480 and SW620 Inositol hexaphosphate (IP6) is molecule, which is usually present in nature, especially in grain and legumes. It has been described several processes, which are influenced in human body by IP6, such as decrease of platelet aggregation, promoting effect on insulin secretion, hypocholesterolemic, as well as anticancer effect. Aim of this work was to study adhesion and migration properties, which are the most important processes in metastasis of cancer cells. For experiments, colorectal adenocarcinoma cell line SW480 and metastatic lymph node cancer cell line SW620, both isolated from identical patient, have been selected. The ability of migration in both cell lines was tested. The influence of IP6 in different concentrations and several time intervals on mRNA (RT-PCR) and protein level (western blot, immunofluorescence) expressions of adhesion and invasion molecules has been compared as well. Considerable ability of migration was found in metastatic cell line...
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Differences in expression of selected microRNA in sensitive and resistant nematodes
Hegrová, Lenka ; Skálová, Lenka (advisor) ; Jirkovská, Anna (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Hegrová Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Differences in expression of selected microRNA in sensitive and resistant nematodes Anthelmintic resistance has become a worldwide growing problem in veterinary medicine due to excessive and inappropriate drug use. Understanding the mechanisms of resistance development could help prevent the spread of resistance and effectively fight resistant individuals. Haemonchus contortus is the most extensively studied parasitic nematode in this regard, because it has the ability to develop resistance to all major classes of anthelmintic drugs and at the same time it has good biological and physiological aspects. Because several recent studies have shown that a change in the expression or activity of so-called microRNAs (miRNAs), small non-coding ribonucleic acid molecules, is also involved in the drug resistance, were monitored in this diploma thesis the differences in the expression of nine selected miRNAs between susceptible (ISE) and multidrug- resistant (WR) strains of H. contortus using the qPCR method. Several significant differences in the expression of selected miRNAs were found between ISE and WR adults....
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The effects and metabolism of anthelmintics in fodder plants
Nováková, Anežka ; Skálová, Lenka (advisor) ; Karlíčková, Jana (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Anežka Nováková Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Consultant: Ing. Lenka Langhansová, Ph.D. Title of diploma thesis: The effects and metabolism of anthelmintics in fodder plants Drugs are potentially dangerous contaminants of the environment. The entry of anthelmintics into the environment is mainly via excrement of treated animals. Anthelmintics can negatively affect the ecosystem by acting on non-target organisms such as plants and soil microorganisms. The accumulation of medicines in plants is also at risk. The consumption of these plants may promote the development of resistance in helminths. This thesis looked at the study of the influence and biotransformation of frequently used anthelmintics of albendazole (ABZ), ivermectin (IVM) and monepantel (MOP) in the forage plants of alfalfa seté (Medicago sativa) and the meadow clover (Trifolium pratense). Plants cultivated in in vitro conditions and plants grown in a greenhouse were used for evaluation. Phytotoxicity was monitored on seeds and whole plants by measuring the length of the roots, by determining the content of proline, light-collecting pigments and the total protein content. Mild phytotoxicity was observed after ivm...
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Laboratory markers in acute heart attack
Dvořáková, Kristýna ; Skálová, Lenka (advisor) ; Dršata, Jaroslav (referee)
Charles university, Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Study program: Healthcare bioanalytics Candidate: Kristýna Dvořáková Thesis supervizor: prof. RNDr. Lenka Skálová, Ph.D. Consultant: Mgr. Martina Hrdinová Title of bachelor thesis: Laboratory markers in the diagnosis of acute myocardial infarction The basic laboratory examination in the diagnosis of acute myocardial infarction (AIM) is primarily the determination of cardiac troponins. The object of this work was to create an overview of markers used in the diagnosis of AIM and to determine cardiac troponins T and I by highly sensitive methods on two different automatic analyzers cobas e 801 and Alinity i in the laboratory of clinical biochemistry. We used diagnostic kits from two different manufacturers to analyze troponins. Cardiac troponins T and I are determined by highly sensitive chemiluminescence methods, which make it possible to capture the increase in concentrations at an early stage of myocardial disability. I compared and evaluated the measured values of troponins in samples from a selected group of patients, in which men aged between 51-70 years predominated. This group is most at risk in the diagnosis of AIM. I made graphical and statistical processing of results separately for men and women....
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Degradation of hyaluronic acid in the skin and percutaneous absorption of exogenous hyaluronic acid through the skin
Exnerová, Andrea ; Skálová, Lenka (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Andrea Exnerová Supervisor: Mgr. Petra Žádníková, Ph.D. prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Degradation of hyaluronic acid in the skin and percutaneous absorption of exogenous hyaluronic acid through the skin Up to half of the body's total hyaluronic acid (HA) content is present in the skin. HA acts as an organizer of the extracellular matrix and is involved in maintaining skin hydration and elasticity. In the last decade, several important findings have been added to the field of HA degradation in the skin, and one of the aims of this thesis was to create a comprehensive overview. Degradation of HA in the dermis was elucidated by the discovery of the essential role in the degradation of HA in dermal fibroblasts played by the KIAA1199 protein. In the epidermis, the lysosomal enzyme hyaluronidase 1 is overexpressed in the stratum granulosum just below the stratum corneum (SC). It is uncertain how HA is internalized into the keratinocytes or whether only intracellular HA is degraded in this way. Epidermal degradation of HA remains elusive. HA is part of a range of topically applied products on the skin, mainly due to its unique moisturizing properties, but it is not yet...
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Changes in expression of biotransformation enzymes in nematodes affected by albendazole
Štěrbová, Karolína ; Skálová, Lenka (advisor) ; Trejtnar, František (referee)
5 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Karolína Štěrbová Supervisor: prof. RNDr. Lenka Skálová Ph.D. Title of diploma thesis: Changes in expression of biotransformation enzymes in nematodes affected by albendazole Barber's pole worm (Haemonchus contortus) is highly pathogenic parasitic nematode, which assaults gastrointestinal system of small ruminants. Disease. Which caused is named by the agent haemonchoses. Long-term infection leads to develop of anaemia, lose weight of host and often even death. For treatment of this disease, synthetic antiparasitic drugs called anthelmintics are used. However, worldwide problem is growth of resistance against all used anthelmintics. Cause of this problem is probably common and unreasonable using of anthelmintics and the exposure of the parasites to sub-lethal doses. This thesis is focused on study of the mechanisms of resistance of barber's pole worm to albendazole, anthelmintic belonging to benzimidazoles group. The increased expression of biotransformation enzymes could contribute to resistance development. In our study, the influence of albendazole on expression of selected genes in adults of H. contortus from the ISE (Inbread Susceptible Edinburgh) strain was studied. Worms...
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Metabolism of flubendazole in human liver subcellular fractions
Holubová, Jana ; Skálová, Lenka (advisor) ; Vokřál, Ivan (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Jana Holubová Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: Metabolism of flubendazole in human liver subcellular fractions Flubendazole (FLU), a benzimidazole anthelmintic drug with a broad spectrum of activity and low toxicity has been used in veterinary as well as human medicine for a long time. Owing its mechanism of action, based on the specific binding to tubulin, FLU is now considered a promising anti-cancer agent. However, we do not have enough information about its biotransformation in human. In our project, subcellular fractions from the liver of 12 human patients (6 males and 6 females) were used to study the stereospecificity, cellular localization, coenzyme preference and possible inter-individual or sex differences in FLU reduction. In addition, the risk of FLU interaction with other drugs was evaluated. Results showed that FLU is predominantly reduced in cytosol and the NADPH coenzyme is preferred. The strict stereospecificity of FLU carbonyl reduction was proven and carbonyl reductase 1 was identified as the main enzyme of FLU reduction in the human liver. A higher reduction of FLU and a higher level of carbonyl reductase 1 protein was found in males than in...
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The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs
Pasák, Michael ; Skálová, Lenka (advisor) ; Trejtnar, František (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michael Pasák Supervisor: prof. RNDr. Lenka Skálová, Ph.D. Title of diploma thesis: The changes in expression of selected drug-metabolizing enzymes in parasites after exposition to sublethal doses of drugs The parasite, which causes significant losses of small ruminants through a disease called haemonchosis, is the abomasal nematode Haemonchus contortus. IVM is one of the anthelmintics used in veterinary medicine to eliminate haemonchosis. The main problem with this parasite is the presence of drug resistance to almost all administered anthelmintics including IVM. It is believed that xenobiotic-metabolizing enzymes such as cytochromes P450 (CYPs) and membrane efflux transporters P- glycoproteins (P-gps), play the role in resistance in H. contortus. This study focuses on the effect of sublethal doses of IVM on expression of selected CYPs and P-gps. Females and males of susceptible ISE strain H. contortus were separated before our experiment. After incubation of nematodes with three different concentrations of IVM, RNA was isolated and the changes in expression of selected genes were analyzed using qPCR. Significant sex-differences were observed in inducibility of tested genes. Significant...
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Study of expression of glutathione peroxidases in mice with induced obesity
Zemanová, Kateřina ; Matoušková, Petra (advisor) ; Skálová, Lenka (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Kateřina Zemanová Supervisor: Ing. Petra Matoušková, PhD. Title of thesis: Study of expression of glutathione peroxidases in mice with induced obesity Obesity is serious worldwide health issue. The most common reason for developing obesity is unbalanced intake and energy expenditure. It is connected to a higher amount of reactive oxygen species (ROS) in organism, which increases the risk of developing health issues related to obesity. Between these issues are belonging mainly cardiovascular diseases, metabolic syndrome or type two diabetes mellitus. The goals of many studies focusing on obesity are identification of new biomarkers, searching for target structures for obesity treatment and potentially developing antiobesity drugs. Glutathione peroxidases (GPxs) are enzymes with antioxidation activity. Currently, there are eight GPxs described. They can reduce the amount of reactive oxygen species (ROS) produced in our body. The GPxs can cause damage to the surrounding tissues in case of their accumulation and they are related to many diseases. Based on these findings, efforts have been made to discover how the expression of GPxs genes is regulated and how they are connected to microRNA...
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