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Morphological characteristic of alterations in the striatum induced by neurodegenerative process in the brain
Němečková, Ivana
("Morphological characteristics of alterations in the striatum induced by neurodegenerative process in the brain") Huntington's disease (HD) is an inherited neurodegenerative disorder. Although the cause of HD, i.e. the production of the mutant form of unstable protein huntingtin (mhtt) which contains 40 and more CAG repeats is known, the effective therapy is not yet available. Therefore, the use of animal models is crucial for the study of the pathogenesis of this fatal disorder. To date, there is no suitable experimental model simulating the neurodegenerative process (NDP) developing in the striatum of the human HD brain. Most of rodent models of HD fall into two broad categories - the neurotoxic lesions and genetically engineered models. The primary aim of our study was a comprehensive morphological description of the development of NDP of HD phenotype in the striatum of the rat brain. We compared the progression of NDP in the lesion induced by intrastriatal injection of quinolinic acid (QA) and in rats transgenic for HD. The groups of male rats surviving for 3, 6-7, 14 days, 1, 3, 6, 9 and 12 months after the QA lesion were compared with 2-, 6-, 12-, 18-, 22-24-month-old tgHD rats and age-matched control (intact) counterparts in both groups. The primary morphological feature of the NDP of HD...
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Immunochemical Detection of PECAM-1 Expression in Hypertensive Rats
Hocková, Erika ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
6 ABSTRACT Charles University in Prague, Faculty of Pharmacy in Hradec Kralove Department of Pharmacology and Toxicology Candidate: Mgr. Erika Hocková Consultant: Doc. PharmDr. Petr Nachtigal, Ph.D. Title of Thesis: Immunochemical detection of PECAM-1 expression in hypertensive rats Hypertension - abnormally high blood pressure - is a lifestyle disease, which belongs to the most frequent cardiovascular diseases. Hypertension seems to be a non-infectious epidemy with 25 to 30 % prevalence in adult population and presents a serious health problem. PECAM-1 is an adhesive signal molecule expressed on hematopoetic and endothelial cells, creating the main component of the vascular tissue. Regarding its structure, it belongs to the immunoglobuline super-family of adhesive molecules. Sunitinib is a biological treatment molecule of the group of tyrosine-kinase inhibitors used for treatment of generalised renal carcinoma and gastrointestinal stromal tumours. Farmacologically, sunitinib is antineoplastic substance - protein-kinase inhibitor. The main adverse effect of sunitinib is hypertension. The aim of this rigorous thesis was to define and to describe the expression of endothelial PECAM-1 in the aorta of spontaneously hypertensive rats (SHR) and their normotensive control group (Wistar - Kyoto rats - WKY)...
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Morphological characteristics of alterations in the striatum induced by neurodegenerative process in the brain
Němečková, Ivana ; Mazurová, Yvona (advisor) ; Anděrová, Miroslava (referee) ; Martínek, Jindřich (referee)
("Morphological characteristics of alterations in the striatum induced by neurodegenerative process in the brain") Huntington's disease (HD) is an inherited neurodegenerative disorder. Although the cause of HD, i.e. the production of the mutant form of unstable protein huntingtin (mhtt) which contains 40 and more CAG repeats is known, the effective therapy is not yet available. Therefore, the use of animal models is crucial for the study of the pathogenesis of this fatal disorder. To date, there is no suitable experimental model simulating the neurodegenerative process (NDP) developing in the striatum of the human HD brain. Most of rodent models of HD fall into two broad categories - the neurotoxic lesions and genetically engineered models. The primary aim of our study was a comprehensive morphological description of the development of NDP of HD phenotype in the striatum of the rat brain. We compared the progression of NDP in the lesion induced by intrastriatal injection of quinolinic acid (QA) and in rats transgenic for HD. The groups of male rats surviving for 3, 6-7, 14 days, 1, 3, 6, 9 and 12 months after the QA lesion were compared with 2-, 6-, 12-, 18-, 22-24-month-old tgHD rats and age-matched control (intact) counterparts in both groups. The primary morphological feature of the NDP of HD...
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Evaluation of DNA damage and repair in various pathological stages
Tenorová, Dana ; Němečková, Ivana (advisor) ; Rathouská, Jana (referee)
author: Dana Tenorová title: Evaluation of DNA damage and repair in various pathological stages Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové field of study: Specialist in Laboratory Methods Chronic obstructive pulmonary disease (COPD) is a serious disease characterized by airflow obstruction that is caused by chronic bronchitis or pulmonary emphysema. As a result of these changes occurs in the organism the oxidative stress with subsequent damage to various biomolecules, which can be measured using a wide range of methods. The primary aim of our study was the evaluation of DNA damage in patients with COPD and comparison to the damage found in the healthy control group. We used electrophoretic method comet assay and we analyzed various types of DNA damage - DNA single strand breaks (SSB), oxidized purines and oxidized pirimidines. We used two forms of this method, i.e. a classical alkaline comet assay and its enzymatic modification, where we worked with endonuclease III (Endo III) and formamidopyrimidin-DNA-glycosylase-III (FPG). Our results showed the significantly higher levels of DNA damage in patients with COPD compared with the control group, especially when using the modified comet assay. In conclusion, we have examined the possibility of comet assay...
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Detection of nestin in tissues
Vařejčková, Michala ; Kudláčková, Zděnka (advisor) ; Němečková, Ivana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Michala Vařejčková Supervisor: PhDr. Zdeňka Kudláčková, Ph.D. Consultant: prof. MUDr. Jaroslav Mokrý, Ph.D. Title of thesis: Detection of nestin in tissues Aim of the work: The aim of this study was to use immunohistochemical methods to show intermediate filament nestin occurs not only in the capillaries but also in large blood vessels - arteries and veins - which has not been published so far. We also investigated coexpression of nestin with a proliferating marker, PCNA. Methods: All studies were performed using embryos 11 and 16 days old from Wistar rats (VELAZ, Prague, Czech Republic). Expression of nestin and PCNA was detected by various types of indirect immunohistochemical method. To amplify a specific signal, we used EnVision system or biotinylated tyramide. Main results: Endothelium of blood vessels expressed nestin with a varying intensity. In some blood vessels expression of nestin was missing. In a developing tissue, nestin is gradually being replaced by increased expression of another type of intermediate filament. To identify tissue antigens that are expressed in a weak intensity, it is better to use signal amplification with tyramide. Conclusion: The results of this study...
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Sunitinib effects on endothelial expression of hemeoxygenase-I in normotensive rats
Svobodová, Barbora ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Barbora Svobodová Sunitinib effects on endothelial expression of hemeoxygenase-I in normotensive rats. Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Medical Laboratory Technician Background: The aim of this bachelor thesis was to define and describe the expression of hemeoxygenase-1(HO-1) in the aortas of normotensive Wistar Kyoto (WKY) rats and evaluate possible effects of sunitinib treatment by means of immunohistochemical and stereological analysis. Methods: Two groups of WKY rats were used for the experiment (6 animals in each of them). Sunitinib was administered to the first group. The other one (the control one) received water only. Application of sunitinib proceeded by following scheme: 8 weeks of using/ 5 weeks rest period/ 2 weeks of using. After that we performed the immunohistochemical analysis on 60 preparations of rat's aortas which observed the expression of HO-1 by methodology En Vision with DAB detection. Then the stereological evaluation was performed. Results: The immunohistochemical analysis showed the expression of HO-1 in the endothelium only and it was noticed just in several animals especially from the group with sunitinib treatment. Conclusion: The immunohistochemical and stereological analysis demonstrated a significant increase of the...
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The effect of atorvastatin on the expression of markers of the cascade transforming growth factor beta in experimental atherogenesis.
Frnochová, Lenka ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Lenka Frnochová The effect of atorvastatin on the expression of markers of the cascade transforming growth factor beta in experimental atherogenesis. Diploma thesis Charles Univerzity in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: The aim of this diploma thesis was to observed the expression of ALK-1 and VEGF in atherosclerotic plaques in ApoE and LDL receptor- deficient mice with regard to the use of atorvastatin. Methods: We used for research female C57BL/6J mice with double deficiency of apolipoprotein E and LDL receptor. Mice were 8 weeks old. Mice were divided into two groups: atorvastatin and control group. Both groups were fed cholesterol diet for 8 weeks and in atorvastatin group was also fed with atorvastatin. Then biochemical analysis of blood was performed. For detection of lipids in atherosclerotic lesions we used histological staining by oil red. Immunohistochemical analysis was performed in 1cm aortic sinus and part of aortic arch. The method used to detect the expression of ALK-1 and VEGF was the avidin-biotin method with detection using DAB. Results: Biochemical analysis showed that atorvastatin reduced levels of total cholesterol. After the staining by oil red atherosclerotic lesions were seen in both groups. Atorvastatin reduced the size of atherosclerotic...
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The study oxidative stress markers in animal model of chronic colitis in large intestine.
Hocková, Erika ; Němečková, Ivana (referee) ; Nachtigal, Petr (advisor)
Crohn's disease and ulcerative colitis are both entities of inflammatory bowel disease (IBD). The nuclear transcription factor kappaB (NFκB) family consists of five different members, which are namely p65 (RelA), c-Rel, RelB, p50 and p52. NFκB has ability to promote the expression of various proinflammatory genes, to influence the course of mucosal inflammation and to mediate different cell-type specific effects. Bilirubin is an endogenous substance with anti-oxidative and anti-inflammatory effects, which influences the process of inflammation and has protective effect. This diploma thesis was aimed at conduction of methods of NFκB detection on paraffine tissue slices and observation of NFκB expression in large intestine walls of normobilirubin and hyperbilirubin rats (Gunn's variety) after chronic imunohistochemically- induced inflammation. Eight Gunn's variety rats (G+ ) and eight Wistar's variety rats (K+ ) were used in this study, all after long time administration of dextrane-sulfate (DSS). For seven following days, DSS (in concentration of 2%w/v) was added into drinkable water for these rats, followed by 14 days of clear drinkable water (without DSS) administered to rats. This three-weeks long cycle was repeated twice (3 cycles in total were completed). Six rats of both varieties (6 G+ and 6...
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