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Development of 3D spheroid cell culture derived from tumor cell lines suitable for photodynamic therapy research
Brieslingerová, Lenka ; Macháček, Miloslav (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Lenka Brieslingerová Supervisor: RNDr. Miloslav Macháček, Ph.D. Title od diploma thesis: Development of 3D spheroid cell culture derived from tumor cell lines suitable for photodynamic therapy research Growing number of tumorous diseases worldwide is urging great effort to make anticancer treatment more effective. Conventional testing of newly developed drugs is performed on tumor cell cultures growing in a monolayer. However, the use of 3D spheroid models provides a more appropriate method of testing, as these models are able to better mimic the in vivo properties of tumors and thus provide a more realistic response to administered drugs. In addition to testing new cytostatics, 3D spheroid models can also be used as tumor models to study the effect of photodynamic therapy (PDT), although this model is still not widely utilized in the field of PDT research. PDT is an alternative and minimally invasive treatment method used not only to treat tumors. It utilises a photosensitizer (PS), molecular oxygen (3O2) and light to form reactive oxygen species (ROS), consequently damaging tumor tissue. This diploma thesis deals with the production of 3D spheroid cell cultures from the HeLa cell line, which...
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Modulation of expression and activity of selected plant detoxifying enzymes by anthelmintics
Graňáková, Patrícia ; Szotáková, Barbora (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Patrícia Graňáková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Consultant: RNDr. Radka Podlipná, Ph.D. Title of diploma thesis: Modulation of expression and activity of selected plant detoxifying enzymes by anthelmintic Anthelmintics represent risk to environment as they may impact non-target organisms including plants, which come into contact with these pharmaceuticals in fields by fertilization with dung from treated animals or in pastures by excrements of treated animals. After uptake, these substances can increase the production of reactive oxygen species in plants, with the risk of oxidative stress and plant damage, and also affect the antioxidant enzymes. The aim of this work was to investigate the effect of two widely used anthelmintics ivermectin and fenbendazole on the activity and expression of selected antioxidant enzymes in soybean (Glycine max). Soybean was cultivated in a greenhouse and watered with a 10 µM solution of the selected drug. The changes of activity and gene expression of antioxidant enzymes were measured in root, leaf, pod and seed samples. Results showed that both anthelmintics caused significant decrease of superoxide dismutase, ascorbate peroxidase, glutathione...
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Molecular mechanisms of cancer cells resistance to anticancer therapy
Koropecká, Magdalena ; Boušová, Iva (advisor) ; Ambrož, Martin (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Science Title, Name, Surname of candidate: Magdalena Koropecká Title, Name, Surname of tutor: doc. PharmDr. Iva Boušová, Ph.D. Title of Bachelor thesis: Molecular mechanisms of cancer cells resistance to anticancer therapy Tumor resistance is still the most common reason for the failure of anticancer therapy. The presented bachelor thesis serves as a summary of professional articles dealing with individual mechanisms of resistance of cancer cells to anticancer therapy. Selected mechanisms are described in detail, namely drug inactivation, alteration of drug targets, decreased drug uptake into the cell, increased drug efflux from the cell, repair of damaged DNA, inhibition of the cell death, epithelial-mesenchymal transition and metastasis, and epigenetic effects. Studying and understanding these mechanisms taking place in tumor cells is crucial for selecting the right treatment and for its success.
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Relationship between compensatory status and renal function in patients with diabetes mellitus type II
Vondráčková, Helena ; Szotáková, Barbora (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Bc. Helena Vondráčková Supervisor: prof. Ing. Barbora Szotáková, Ph.D. Specialist consultant: MUDr. Pavel Pick Title of diploma thesis: Relationship between compensatory status and renal function in patients with diabetes mellitus type II Diabetes mellitus is a chronic disease accompanied by a number of complications. Among the most common and at the same time the most serious are kidney diseases, which often lead to their failure. These patients then rely on lifelong dialysis treatment or a possible kidney transplant. By compensating for diabetes mellitus effectively, the development of complications can be reduced or at least delayed. Fasting and postprandial blood glucose level and glycated hemoglobin are used to assess the state of compensation. The renal function is assessed by the examination of albumin and proteins in urine, serum creatinine concentration and estimation of glomerular filtration. The aim of the thesis was to determine whether there is a relation between the level of compensation for diabetes mellitus and the renal function. The data from 70 patients diagnosed with diabetes mellitus type II regularly checked in a diabetological clinic were evaluated. Samples were...
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Effect of selected bicyclic monoterpenes on the activity and expression of antioxidant enzymes in human liver
Křížová, Anna ; Boušová, Iva (advisor) ; Szotáková, Barbora (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Anna Křížová Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Title of diploma thesis: Effect of selected bicyclic monoterpenes on the activity and expression of antioxidant enzymes in human liver Monoterpenes are plant secondary metabolites, which are used as flavourings or aromas in the food industry due to their smell and flavour. They also play an irreplaceable role in the cosmetic and pharmaceutical industry. Monoterpenes are substances with a wide spectrum of biological activities. They exert, for example, antimicrobial, anti- inflammatory, antioxidant, hypotensive, and analgesic effects. In addition to the positive effects of these substances, some cases of their toxic effects in various organs were also observed, mostly in the liver. Monoterpenes also affect the activity and expression of antioxidant enzymes. The aim of this thesis was to determine the effect of five selected bicyclic monoterpenes, (+)-camphor, (-)-camphor, (-)-fenchone, camphene, and (-)-α-thujone, on the activity and expression of antioxidant enzymes. Of those enzymes, glutathione-S-transferase (GST), glutathione peroxidase (GPx), glutathione reductase (GR), superoxide dismutase (SOD), and catalase (CAT) were studied. At...
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Antiproliferative and anthelmintic effects of fern extracts
Turoňová, Petra ; Matoušková, Petra (advisor) ; Boušová, Iva (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Bichemical Sciences Candidate: Petra Turoňová Supervisor: doc. Ing. Petra Matoušková, PhD. Title of diploma thesis: Antiproliferative and anthelmintic effects of fern extracts Ferns belong to spore vascular plants. In this study, an antiproliferative and anthelmintic effects of selected fern extracts were investigated. The antiproliferative effect of the extracts was tested on the SW480 cell line. This cell line was originally obtained from primary adenocarcinoma of the colon. The anthelmintic effect was tested on L3 larvae Haemonchus contortus, the most common internal parasite of sheep and goats. Firstly, to determine the antiproliferative effect of the extracts, an initial screening was performed. Cytotoxicity was evaluated by measuring absorbance on a Tecan spectrophotometer. The four fern species that showed most promising antiproliferative activity were tested at three time intervals and four concetrations. H. contortus larvae L3 were exposed to fern extracts for seven days and the percentage of developed L4 larvae was detected by microscope. Athyrium distentifolium, Dryopteris aemula, Davallia canariensis and Polystichum aeculatum showed the highest antiproliferative and anthelmintic activity. In addition, Dryopteris...
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ln vitro characterization of acetylcholinesterase modulators
Múčková, Ľubica ; Jun, Daniel (advisor) ; Boušová, Iva (referee) ; Štětina, Rudolf (referee)
Various in vitro approaches are available to evaluate the toxicity of substances as well as their mechanism of toxicity in the early stages of the process of developing new chemical entities. These systems are mainly used for screening and allow the creation of more complex toxicological profiles. The present work focuses on in vitro evaluation of substances modulating acetylcholinesterase activity. The enzyme plays a crucial role in the nervous system and occurs mainly in cholinergic synapses and neuromuscular junctions. The primary biological function of this enzyme is the termination of the nerve impulse at these synapses by the rapid hydrolysis of the neurotransmitter acetylcholine to choline and acetate. Acetylcholinesterase is a target enzyme for many drugs, e.g., for Alzheimer's disease or other neurodegenerative diseases. The most often used are reversible and pseudoreversible inhibitors. It also plays a role in poisoning by chemical warfare agents, namely nerve agents and organophosphorus pesticides. Acetylcholinesterase reactivators are used as antidotes for these poisons. Reactivators (asoxime, pralidoxime, obidoxime, trimedoxime, methoxime and oximes K027, K048, K074, K075, and K203) and inhibitors (pyridostigmine, galantamine, rivastigmine, donepezil, tacrine, 7-methoxytacrine, and...
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In vitro effects of 3-hydroxytyrosol on renal hypoxia and inflammation
Kamasová, Terézia ; Boušová, Iva (advisor) ; Macháček, Miloslav (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Terézia Kamasová Supervisor: Asst. Prof. PharmDr. Iva Boušová, Ph.D. Prof. Alice Santos-Silva Maria João Valente, Ph.D. Title of diploma thesis: In vitro effects of 3-hydroxytyrosol on renal hypoxia and inflammation Chronic kidney disease (CKD) results from a group of heterogeneous disorders affecting the kidneys. The renal hypoxia and hypoxia-derived oxidative stress, renal fibrosis, and inflammation are highly prevailing conditions appearing in the diseased kidney, contributing to the progression of CKD. Phytochemicals are an essential part of contemporary therapeutic strategies for the treatment of various diseases. 3-Hydroxytyrosol (HT), a phenolic compound extracted from olives and olive-derived products (e.g. olive oil), is believed to carry a potent antioxidant, anti-inflammatory, antithrombotic, bactericidal and bacteriostatic activity. The aim of this work was to determine the preventive effect of HT in hypoxic renal cells and evaluate the effect of HT on hypoxia-related inflammation, fibrosis, and oxidative stress, in order to summarize the value of this phenolic compound as a promising novel remedy in the treatment of CKD. A cell line of human renal proximal tubular cells (HK-2) was...
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Effect of camphor isomers on the expression of drug-metabolizing enzymes in human liver cells
Smolíková, Michaela ; Boušová, Iva (advisor) ; Ambrož, Martin (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Smolíková, DiS. Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Advisor: Mgr. Michaela Šadibolová Title of diploma thesis: Effect of camphor isomers on the expression of drug-metabolizing enzymes in human liver cells Camphor is a cyclic ketone from the group of monoterpenes. Two camphor isomers occur in nature. Currently, camphor is used as an antiseptic, analgesic, rubefacients, and anti- inflammatorydrug. Camphor is sed mainly in preparations for topical use, where it has a mild anesthetic effect and a feelingof warmth together with its characteristicallystrong fresh scent. Most camphor intoxications are caused by improper use or dosing. The aim of the diploma thesis was to determine the effect of isomers (+)-camphor and (-)-camphor on the expression of selected phase I and II biotransformation enzymes. The precision-cut human liver slices prepared from a total of three patients, two men and one woman at the age range of 46 - 49 years,were used as biological material.Liver slices were incubatedwith10 µM and 50 µM (+)- camphor and (-)-camphor for 24hours at 37řC. Proteinexpressionof phase I (cytochrome P450 (CYP) 3A4, CYP2C, aldo-keto reductase (AKR) 1C3), and phase II enzymes...
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Anticancer activity of semisynthetic prenylflavonoids in SW480 cell line
Blahová, Kateřina ; Boušová, Iva (advisor) ; Skarková, Veronika (referee)
Charles University Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Kateřina Blahová Supervisor: doc. PharmDr. Iva Boušová, Ph.D. Advisor: PharmDr. Martin Ambrož, Ph.D. Title of diploma thesis: Anticancer activity of semisynthetic prenylflavonoids in SW480 cell line Cancer belongs to the leading causes of mortality in developed countries. Colorectal carcinoma is the second most common cancer in the Czech Republic. Due to the development of resistance to classical chemotherapeutics, a search for new treatment strategies is ongoing. Prenylflavonoids belong to the natural compounds, which express anticancer effect, with xanthohumol and prenylated derivatives of naringenin belonging to the often-studied compounds. In this diploma thesis, anticancer properties of naringenin and its five semisynthetic prenylated derivates were tested in the cell line SW480, which is derived from colorectal carcinoma. Cell viability was monitored by the neutral red uptake test after 72 h treatment. For compounds with marked anticancer activity, the value of IC50 was determined and the effect of those compounds on the cell cycle was determined by flow cytometry. Substantial antiproliferative effect was found in four compounds (derivatives A, B, C, and E), values of IC50 for these...
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