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Therapy of hematologic cancer
Baniková, Lucia ; Štaud, František (advisor) ; Čečková, Martina (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Lucia Baniková Supervisor: Doc. PharmDr. František Štaud, PhD. Title of diploma thesis: Therapy of hematological malignancies Hematologic malignancies are cancers of the blood, bone marrow or lymph nodes. Individual hematological malignancies are defined and distinguished from one another essentially according to four parameters: clinical features, microscopic morphology, immunophenotype and molecular/genetic features. Despite the progress made in the last decade in the treatment of haematological malignancies, most of the patients still have a dismal prognosis. However, the improved knowledge of tumour biology opened the possibility to develop new 'intelligent' therapeutic strategies, the so-named targeted therapies. These approaches aim to selectively kill cancer cells by basing this selectivity on both the expression of a specific molecule on their surface or the activation of particular molecular pathways secondary to malignant transformation. Extensive research has led to the understanding that there are many more subtypes of haematological malignancies. Each of these subtypes behaves in its own particular way and requires its own particular therapy. Correct treatment depends...
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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...
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Investigating the role of zinc transporter ZIP 6 and STAT3 in mitosis
Burgetová, Lenka ; Čečková, Martina (advisor) ; Štaud, František (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Student: Lenka Burgetová Supervisor: PharmDr. Martina Čečková PhD. Specialized supervisor: Dr. Kathryn Taylor PhD. Title of diploma thesis: Investigating the role of zinc transporter ZIP 6 and STAT3 in mitosis It has been shown that STAT3 (signal transducer and activator of transcription 3) plays a role in the development of cancer. ZIP6 is the downstream target of this transcription factor. Previous research has focused mainly on the activation of STAT3 by tyrosine phosphorylation, while the effect of phosphorylation at a second site, serine 727, remained relatively uninvestigated. In this study, it is proposed that serine-phosphorylated STAT3 is activated throughout mitosis in tamoxifen-resistant (TamR) breast cancer cells and that zinc transporter ZIP6 and serine-phosphorylated STAT3 are involved in a zinc-mediated mitotic mechanism. After using nocodazole to induce mitotic arrest, the expression of tyrosine- phosphorylated STAT3 protein was observed to be reduced while the expression of serine- phosphorylated STAT3 was increased. Zinc supplementation after nocodazole treatment appeared to push cells through mitotic-arrest and cause proteolytic cleavage of STAT3 suggesting a novel...
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Genotypizace ApoE u dětí
Weinfurterová, Radka ; Štaud, František (advisor) ; Nachtigal, Petr (referee)
The purpose of this study was to find out how genotype of apolipoprotein E influences blood levels of LDL cholesterol, HDL cholesterol and triglycerides in children. In our study there was found out that apo E genotype has a significant influence on apo E levels, total cholesterol levels and levels of LDL and HDL cholesterol. Subjects with allele apo E4 had significantly higher levels of LDL cholesterol and subjects who carried allele E2 had conversely lower level of LDL cholesterol. These outcomes corresponded with previously made studies.
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Preparation of recombinant cDNA of drug transporters
Hadrabová, Zdeňka ; Štaud, František (advisor) ; Červený, Lukáš (referee)
We have prepared suitable plasmid containing coding sequence of MRP3, for subsequent preparation of cell line overexpressing this transporter. Composition of reaction mixture and temperature profile of PCR reaction for MRP3 coding sequence amplification was determined experimentally. Adenosine was added to 3' ends of PCR product to allow cloning into PCR® -XL-TOPO® vector. pCR® -XL-TOPO® vector containing cloned sequence and pZeoSV2(-) plasmid were cleaved using restriction endonucleases EcoRV and SpeI. Afterwards it was possible to clone cleaved coding sequence into pZeoSV2(-) plasmid due to complementary ends. Prepared plasmid is suitable for transfection of eukaryotic cell lines. The cloning was successful.
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Pharmacological influence of endothelial dysfunction in experimental model of diabetic rat
Přidalová, Hana ; Nachtigal, Petr (advisor) ; Štaud, František (referee)
The aim of this thesis was to evaluate the expression of cell adhesion molecules in femoral artery after induction of diabetes and after the administration of sulodexide. We focused on the expression of PECAM-1, ICAM-1 and VCAM-1 vessel endothelium. The analysis was provided by means of immnohistochemical staining. Male Wistar rats were divided into three groups. Control group received saline, the second group received sulodexide (SLX, 100 UI/kg/den). Diabetes was induced by the streptozotocine injections (30mg/kg/den i.p.) during consecutive free days. The treatment was administered for 10 weeks. Biochemical results confirmed the induction of diabetes; however the suledoxide treatment did not decrease glucose levels when compared with non-treated diabetic animals. The ICAM-1 expression was slightly increased in diabetic treated rats when compared with control rats however suledoxide treatment did not affect ICAM-1 expression. VCAM-1 expression was not detected in any rat from control, diabetes and/or suledoxide treated rats. In conclusion the results of this pilot study showed that diabetes induced by the administration of streptozotocine did not resulted in significant induction of endothelial dysfunction. Thus the possible effect of suledoxide treatment could not be evaluated in this study.
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The oxidative stress in anthracycline cardiotoxicity in rabbit
Dubská, Veronika ; Nachtigal, Petr (advisor) ; Štaud, František (referee)
The aim of this thesis was to evaluate possible changes of 4-hydroxynonenal expression in rabbit aorta after repeated administration of daunorubicin. Chronic anthracycline cardiotoxicity was induced by repeated administration of daunorubicinu (3 mg/kg=50 mg/m2 i.v., 1x week) for the period of 10 weeks. We focused on the monitoring VCAM-1 and ICAM-1 expression in rabbit aorta. Ten daunorubicin groups were compared with control rabbits. The animals were killed 24 hours and/or 7 days after the administration of daunorubicin. Immunohistochemical analysis showed no expression of 4-HNE in any control or experimental groups. In conclusion, the administration of daunorubicin did not induce 4-HNE expression in aorta in either control and daunorubicin treated rabbits, suggesting that endothelial dysfunction and/or oxidative stress in aorta is not triggered by daunorubicin treatment in vivo.
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Monitoring the effect of atorvastatin on the expression of endoglin / / and endothelial NO synthase (eNOS) in mouse model of atherosclerosis
Uhlárová, Iva ; Nachtigal, Petr (advisor) ; Štaud, František (referee)
Atherosclerosis, or sclerosis of arteries, is a degenerative disease of arteries. Sometimes it is called "the disease of 20th century". ApoE/LDL - receptor double knockout mice represent a new animal model for study of atherogenesis, which is characterized by severe hyperlipidaemia and atherosclerosis. Statins (or competitive inhibitors 3-hydroxyl-3-methyl-glutaryl-coenzym A-reductase) currently belong to the most efficient and the most useful hypolipidemic drugs for all over the world. They decrease mainly levels of total cholesterol and LDL cholesterol. The aim of this thesis was to describe the expression of endoglin in atherosclerotic plaques in apoE/LDL-receptor deficient mice. Moreover we wanted to determine endoglin colocalization with eNOS and the effect of atorvastatin treatment on the expression of both molecules. ApoE/LDLR-deficient mice on were subdivided into 2 groups. The control group of animals was fed with the western type diet. The same atherogenic diet was used in ATV group, where atorvastatin was added to the atherogenic diet at the dosage of 100 mg/kg per day. The results of this thesis confirmed the expression of endoglin in atherosclerotic lesions in ApoE/LDLR-deficient mice. The expression of endoglin was located on the aortic vascular endothelium and in other smaller...
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Gene silencing of ICAM-1 by lysine modified oligonucleotides
Kocourková, Aneta ; Štaud, František (advisor) ; Nachtigal, Petr (referee)
Conjugation of ligands to antisense oligonucleotides is a promising approach for enhancing their effects on gene expression. In this study 2'-O-lysylaminohexyl group was linked to the uridine base, which replaces one, two or three thymine bases thus modifies the oligonucleotides. This exchange of bases was tested for improvement of silencing target protein expression. Effectivity of modifications in silencing target protein expression was examined with the alicaforsen sequence (DNA) and siRNA. Alicaforsen, currently in clinical trial 3, is a phosphorothioate targeting ICAM-1, which was the model used to evaluate the influence of modifications. The same target was chosen for siRNA to compare the efficiency of DNA and siRNA substances. For the first time, down-regulation of ICAM-1 was shown on the blood brain barrier cell line ECV304. Unmodified/modified antisense oligonucleotides and siRNA sequences were transfected into ECV304 cells with the help of a transfection agent lipofectamine 2000. After 24 hours of transfection cells were disrupted by a chemical lysis. Protein concentrations were determined by Bradford protein assay. ICAM-1 inhibition was assessed with western blot. The inhibitory effect of ICAM-1 was normalized to the corresponding actin and untreated cells. ICAM-1 protein levels were...
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