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Alteration of transport proteins expression during obstructive cholestasis in rats I
Šindelářová, Jana ; Doleželová, Eva (advisor) ; Zemánková, Lenka (referee)
Jana Šindelářová Alteration of transport proteins expression during obstructive cholestasis in rats I Diploma thesis Charles University in Prague, Fakulty of Pharmacy in Hradec Králové Pharmacy Background: The purpose of the diploma thesis was the confirmation of liver cholestatic damage induced by bile duct obstruction lastnig for 28 days by biochemical analysis and analysis of mRNA and protein expression of liver uptake transporters in rats (Ntcp, Oatp1a1, Oatp1a4, Oatp1b2, Oat2). Methods: Wistar rats (n = 6, in each group; 280 - 320g) were dividend in two groups: control group of sham-operated animals (Sham) and group with bile duct obstruction lastnig for 28 days (BDO). Biochemical analysis of serum was performed by Cobas Integra ® 800. Changes of mRNA and protein expression of transport proteins were evaluated by qRT-PCR and Western blot. Results: In BDO group, level of bile acids increased to 454%, level of total bilirubin to 4111% and level of conjugated bilirubin to 7313% as compared to control group. In comparison to control group, activity of ALP was elevated to 238%, activity of GMT to 2826%, activity of ALT to 1200% and activity of AST to 1378%. As compared to Sham group BDO group showed decreased mRNA levels of Ntcp to 51% and of Oatp1a4 to 38%. No signifiant changes were found in...
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Alteration of transport proteins expression during obstructive cholestasis in rats II
Skořepová, Zuzana ; Doleželová, Eva (advisor) ; Němečková, Ivana (referee)
5 Abstract Zuzana Skořepová Alteration of transport proteins expression during obstructive cholestasis in rats II Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: The aim of the diploma thesis was the verification of cholestatic liver impairment induced by bile duct obstruction lasting for 28 days in rats, by analysis of mRNA and protein expression of liver efflux transport proteins (Bsep, Bcrp, Mrp2, Mdr1, Mdr2, Mrp3, Mrp4). Methods: Wistar rats (n = 6, in each group; weighing 280 to 320 g) were divided into two groups: control rats were sham-operated (Sham) and BDO group (bile duct obstruction lasting for 28 days). mRNA and protein expression changes of the transporters were perfomed by qRT-PCR and Western blot. Results: As compared to Sham group, BDO group showed increase of mRNA expression of Mdr2 transporter to 303%, Mdr1b to 340% and Mrp3 to 9900%. Mrp2 mRNA expression was decreased to 42%. Obstructive cholestasis led to significant increase of protein levels - Bcrp to 160%, Mdr1 to 1010%, Mrp3 to 940% and Mrp4 to 140%. Mrp2, Bsep and Mdr2 protein levels were increased to 31%, 59% and 82%, respectively. Conclusion: Obstructive cholestasis led to decrease of canalicular liver transporters expression and increase of basolateral liver transporters...
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The study of VCAM-1 expression in experimental atherogenesis
Mertlíková, Veronika ; Nachtigal, Petr (advisor) ; Doleželová, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate: Veronika Mertlíková Supervisor: Doc. PharmDr. Petr Nachtigal Ph.D. Title of diploma thesis: The study of VCAM-1 expression in experimental atherogenesis. The aim of the thesis is to describe the expression of VCAM-1 in the vessel wall of normocholesterolemic C57BL/6J mice, which were fed standard laboratory diet, and hypercholesterolemic apoE-deficient mice. As an experimental model normocholesterolemic C57BL/6J male mice and apoE-deficient mice at the age of 6 weeks were used. The C57BL/6J male mice were fed standard laboratory diet and apoE-deficient mice were fed cholesterol diet for 8 weeks. After this procedure a biochemical analysis of blood and immunohistochemical analysis of segments taken from aortic sinus and aortal arch were made. In comparison with the control C57BL/6J mice a large level of cholesterol was found in the blood of the apoE-deficient mice. Atherosclerotic lesions are thus generated only by the hypercholesterolemic mice. We were monitoring the expression of VCAM-1 in aortas of the normocholesterolemic C57BL/6J mice without atherosclerotic lesions by immunohistochemical methods, and the same methods were used for monitoring of the areas with and without...
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Immunohistochemical Analysis of Expression of Phosphorylated form Smad2 after Administration of Atorvastatin in Experimental Atherogenesis
Vybulková, Lucie ; Nachtigal, Petr (advisor) ; Doleželová, Eva (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Candidate Mgr.Lucie Vybulková Consultant Doc.PharmDr.Petr Nachtigal, PhD Title of Thesis Immunohistochemical analysis of expression of phosphorylated form Smad2 after administration of atorvastatin in experimental atherogenesis Background: The aim of this thesis was to describe the effects of atorvastatin treatment on the expression of phosphorylated form of Smad2 in atherosclerotic plaques in ApoE/LDLr-deficient mice by immunohistochemical methods. Furthermore, the influence of atorvastatin on lipid spectrum and size of atherosclerotic plaques was observed. Methods: In our experiment there were used C57BL/6J female mice with double deficit of apolipoprotein E and LDL receptor. There was performed the biochemical analysis of blood samples, histological staining with oil red for detection of tissue lipids, quantitative analysis of size of lesions and immunohistochemical analysis of samples containing semilunar valves and aorta for the detection of expression of Smad2. It was done using the method Avidin-Biotin and detection with DAB. Results: Using biochemical analysis we confirmed increased levels of total cholesterol and VLDL after the administration of atorvastatin. Smaller size of...
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The studying of some markers of transforming growth factor pathway in mouse model of atherosclerosis.
Pagurko, Lukáš ; Nachtigal, Petr (advisor) ; Doleželová, Eva (referee)
Abstracts Lukáš Pagurko The studying of some markers of transforming growth factor pathway in mouse model of atherosclerosis. Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: We observed the expression of endothelial nitric oxide synthase (eNOS) in mice atherosclerotic plaque by using imunohistochemical methods. The aim was to prove nonlipid-lowering effects of atorvastatin to affect the expression of eNOS. Methods: Female apoE and LDLr deficient mice, aged 16 weeks were used in the study. Biochemical analysis of blood samples and imunohistochemical analysis of the tissue segments were performed. For identification of eNOS expression was used EnVision method with DAB visualization. Results: We found out in biochemical analysis that 8-weeks doses of atorvastatin significantly increases levels of overall cholesterol. Presence of atherosclerotic lesions was observed at all incisions in control group even in atorvastatin group, whereas aterom necrotic fibres was founded at a number of them. The administration of atorvastatin were lead to significant decrease of atherosclerotic lesions compared to untreated group. There were also rapid increase of eNOS expression especially on the surface of atherosclerotic lesions. Conclusion: The positive influence of...
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Spirulina Platensis effects on atherogenesis in mouse model of atherosclerosis
Žáková, Marie ; Nachtigal, Petr (advisor) ; Doleželová, Eva (referee)
Marie Žáková Spirulina platensis effects on atherogenesis in mouse model of atherosclerosis Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: The aim of this diploma thesis was to evaluate possible antiatherogenic and antiinflammatory effects of Spirulina platensis in apoE-deficient mice. Methods: We used C57BL/6J male mice with deficiency of apolipoprotein E (apoE-/-) at the age of 3 months. The animals were divided into 2 groups (control and examined mice). Both groups were fed with cholesterol diet for 8 weeks. In Spirulina group, Spirulina platensis was added at dose of 20 mg/kg/day. The biochemical analysis was performed in order to discover the lipid spectrum of blood. There was also performed the histological staining with oil red in order to visualise the size of atherosklerotic leisons and to detect the lipids. The aim of the immunohistochemical analysis was to approve the amount of VCAM-1 expression. Results: Biochemical analysis did not demonstrate any significant decrease of total cholesterol expect from decreasing of LDL cholesterol. The presence of atherosclerotic lesions with lipids cumulation was illustrated by histological staining with oil red in both mice groups, the results of each group did not differ. In comparison with...
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The study of transforming growth factor receptor during atherogenesis in mouse model of atherosclerosis.
Vybulková, Lucie ; Nachtigal, Petr (advisor) ; Doleželová, Eva (referee)
Lucie Vybulková The study of transforming growth factor receptor during atherogenesis in mouse model of atherosclerosis Diploma thesis Charles University in Prague, Fakulty of Pharmacy in Hradec Králové Pharmacy Background: The aim of this diploma thesis was the establishment of methodology for detection of TGF-βRI, subtype ALK-5, and studying its expression in the aortic root in apoE/LDLr- deficient mice. Methods: Using female mice C57BL/6J strain with a double deficit of apolipoprotein E and LDL-receptor was performed biochemical analysis of blood specimens, histologic staining with oil red for detection of lipids in tissues and immunohistochemical analysis of segments of tissue from the aorta and the top half of the heart. To detect expression of ALK-5 was used EnVision method with DAB visualization. Results: Using biochemical analysis, we confirmed increased levels of cholesterol in mice fed cholesterol diet as compared with mice on standard diet. At histologic staining with oil red we detected increased formation of atherosclerotic lesions in mice with cholesterol diet. Immunohistochemical analysis of expression of ALK-5 we demonstrated high expression of ALK-5 in the tunica media and in atherosclerotic plaques with no significant difference in the two groups of mice. Conclusion: The results...
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Evaluace www stránek vybraných municipalit
Doleželová, Eva
This paper establishes ground for the redesign of Brno-střed city district web page. For the purpose of this study web pages of several municipalities were evaluated: Brno-střed city district, Nové Město na Moravě, Praha 1 and Plzeň. Study shows how the selected municipalities fulfill the legal regulations of the Act no. 106/1999 on free access to information and the Act no. 81/2006 Coll. which states that municipalities are obligated to provide the information on web pages also to handicapped or disabled. Furthermore the usability analysis and quantitative survey were applied to municipality web page in Brno-střed region to determine the public needs and requirements. Presented findings indicate both significant failure in fulfilling the law and public dissatisfaction with Brno-střed web page content and structure. This dissertation provides sample solutions for web page usability.
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