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Immunohistochemistry as a tool for cell phenotyping
Hajzlerová, Petra ; Němečková, Ivana (advisor) ; Soukup, Tomáš (referee)
10 Abstract Author: Petra Hajzlerová, DiS. Title of thesis: Immunohistochemistry as a tool for cell phenotyping Bachelor thesis Charles University in Prague Faculty of Pharmacy in Hradec Králové Study course: Medical laboratory technician The aim of this study was to provide information about phenotypization of cells in tissues using immunohistochemical analysis. Several immunohistochemical techniques are used for cell phenotypization and to define their advantages. Immunohistochemical technique is based on detection of the tissue/cell antigen using specific antibody. These specific antibodies are most often labeled by enzymes or fluorochromes, less frequently by radionuclides or dense particles. Immunofluorescence uses fluorochromes and enzymatic immunohistochemistry uses enzymes. Immunohistochemical methods can be divided into direct and indirect methods. The most commonly used methods in laboratory practice are indirect methods, especially those based on the interaction between avidin and biotin (LAB and ABC). Methods based on the interactions of enzyme anti-enzyme are used less frequently. The most popular in laboratory practice are methods based on soluble polymer, e.g. EnVision and EPOS. Tyramides are very useful for signal amplification in immunohistochemistry. These methods were compared in this...
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Effect of high soluble endoglin levels on the liver morphology in mice.
Burešová, Petra ; Němečková, Ivana (advisor) ; Kudláčková, Zděnka (referee)
Author: Petra Burešová Title of thesis: Effect of high soluble endoglin levels on the liver morphology in mice Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Study course: Medical laboratory technician Endoglin (CD105) is an integral transmembrane protein with total weight of 180 kDa. It is characterised by its participation in the signaling cascade of transforming growth factor β (TGF-β) due to its role as a co-receptor of this factor. In addition, a soluble form of endoglin (sol-Eng) which is produced by proteolytical sheeding from the extracellular part of membrane bound endoglin into circulation was demonstrated. Recently, it was shown, that endoglin is crucial factor in the development of both, cardiac fibrosis in the failing myocardium and liver fibrosis. The aim of our work was to detect liver fibrosis in transgenic mice. Using basic histological techniques and collagen staining we evaluated development of liver fibrosis in transgenic mice overexpressing human soluble endoglin on chow and high fat diet in comparison with control mice with low levels of human soluble endoglin. Our histological results show that high soluble endoglin levels have no effect on the development of fibrotic changes in the liver. However, it will be very interesting to evaluate...
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C-reactive protein effects on aortic endothelium in spontaneously hypertensive rats
Kosán, Aleš ; Němečková, Ivana (advisor) ; Doleželová, Eva (referee)
C-REACTIVE PROTEIN EFFECTS ON AORTIC ENDOTHELIUM IN SPONTANEOUSLY HYPERTENSIVE RATS Diploma thesis Author: Aleš Kosán Study program: Pharmacy Charles University in Prague Faculty of Pharmacy in Hradec Králové Endothelial dysfunction and chronic inflammation are factors figuring in pathogenesis of metabolic syndrome. This paper is outlined as a pilot study, which represents basis for further research in the question of C-reactive protein influence on endothelium. The expression of four proteins previously associated with the development of endothelial dysfunction, was studied using the Western blot method: endothelin-1, heme oxygenase-1, endoglin and phosphorylated nitric oxide synthase. Analysis was carried out on the samples of homogenized rat aortas of transgenic spontaneously hypertensive rats with expression of human C-reactive protein. A standard spontaneously hypertensive rat strain was used as a control group. The detection was made by chemiluminescence substrate on the X-ray films and data were semiquantitatively evaluated by densitometry. We found increased expression in endoglin levels, valued at 186.3 % contrary to the control group. The expression of phosphorylated nitric oxide synthase was 325.6 % in comparison with the control group. The increase of expression was minimal (109.0 %) in...
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Soluble endoglin effects on vascular endothelium
Ďaďovská, Lucia ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Soluble endoglin effects on vascular endothelium Diploma thesis Lucia Ďaďovská Supervisor: doc. PharmDr. Petr Nachtigal, Ph.D. Background: We observed the effect of soluble endoglin on expression of VCAM-1 and occurrence of macrophages in descendent aorta in murine models. Analysis was performed by immunohistochemical methods with fluorescence detection. Methods: In this study, we used the transgenic mice overexpressing human soluble endoglin (Sol-Eng+) and control mice with low plasma levels. The both groups were the same age and during the entire three months experiment were fed high fat rodent diet. For results analysis, we used tissue sections of descendent aorta and immunohistochemical methods with fluorescence microscopy detection. Results: Immunohistochemistry showed there was not an evidence of macrophage infiltration in tissue samples of descendent aorta in either control mice or in Sol-Eng+ mice. Immunohistochemical analysis of VCAM-1 expression in descendent aorta of control mice and of Sol-Eng+ mice presented no difference in localization and strength of VCAM-1 expression in both groups. Increased VCAM-1 expression was detected at the exactly same location in both groups -...
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Sunitinib Effects on the Expression of ICAM-1 in Normotensive and Hypertensive Rats
Frnochová, Lenka ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Mgr. Lenka Frnochová Sunitinib effects on the expression of ICAM-1 in normotensive and hypertensive rats Thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Pharmacy Background: We quantified the expression of ICAM-1 in aorta of normotensive Wistar Kyoto rats and spontaneously hypertensive rats during use of sunitinib. For quantification we applied stereological and immunohistochemical methods. Methods: We used inbred male of Wistar Kyoto rats and spontaneously hypertensive rats for our research. Rats of both strains were divided into two groups - a sunitinib group and a control group. The experimental group was receiving drinking water with sunitinib of 10 mg/kg per day and rats in the control group was receiving only drinking water. Spontaneously hypertensive rats were receiving sunitinib for 8 weeks and after 5 days of regeneration they got sunitinib into the water back for another 8 weeks. Wistar Kyoto rats were receiving sunitinib in water for 8 weeks, but after 5 days of regeneration, these rats were receiving sunitinib only for 2 weeks due to toxicity and weight loss. Immunohistochemical analysis was applied in rat aorta. For detection of the ICAM-1 expression was used En Vision method with DAB detection. Results: Immunohistochemical staining showed that expression of...
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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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Social stratification of domain Horšovský Týn in the 17th - 19th century
Němečková, Ivana ; Šedivá Koldinská, Marie (advisor) ; Maur, Eduard (referee)
T-Mobile Czech Republic a.s. Abstract.docx 1/1 Abstract This thesis is focused on the living conditions of the serf population in the West Bohemian domain of Horšovský Týn beetween 17th and 19th centuries. The core of this work is in the 17th century. The geografical area of the domain was defined in the Tax Rule - the city, fifty- four villages and five partial villages. This paper consists of four basic sections. In the first section you can find the overview of the main literature regarding this topic. The second one is dedicated to the brief historical and territorial development of the domain and to the characteristic of the administrative and economic circumstances. The demografical structure of the domain in the 17th century is described in depth in the third section. This analysis was done on the basis of the register of the serf people classified according to their religion. The focus is partially dedicated to the social structure as well, mainly to the houseless lodgers and servants. The typology and composition of the households is discussed briefly. Above that, there is an approximation of population in the domain Horšovský Týn from the mid 17th until the end of 19th century. The last section is focused on the social stratification of the domain and its changes from the mid 17th to the mid 19th...
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Impregnation methods - silver stain -their importace in diagnostic and transplant patology
Květoň, Martin ; Semecký, Vladimír (advisor) ; Němečková, Ivana (referee)
Page 1 of 1 Abstract Martin Květoň Impregnation methods - silver stain -their importace in diagnostic and transplant patology Bachelor's thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Medical bioanalytics The impregnating histological methods are based on saturation of tissue sections with metal salts. The most commonly used silver staining methods employ silver ions for impregnation. Product of those methods is elementary silver, reduced "in situ" to the specific tissue components. (Newman, and others, 1998) The aim of this thesis was to analyze the most commonly used impregnating methods, to show their use in diagnostic pathology and to present the methodics of those techniques. The secondary aim of this thesis was to evaluate their importance in routine histopathological practice. The introductions part contains general information about the impregnating methods and their use. General theoretical part deals with the principle of those methods, special part deals with the structures which are showed by using those methods, and with general description of histological process. Methodical part contains detailed descriptions of chosen silvering methods, and their results. Conclusions have shown that the aims of the thesis have been met and that the impregnating methods are an...
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Immunohistochemical detection of macrophages in atherosclerotic lesions brachiocephalic artery I
Škodová, Sabina ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Sabina Škodová Immunohistochemical detection of macrophages in atherosclerotic lesions brachiocephalic artery. Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Medical Laboratory Technician Background: The aim of this bachelor thesis was to set imunohistochemic methods for detection of the macrophages in brachiocephalic artery apoE/LDLR receptor deficient mice. This thesis also deals with quantification of expression macrophages in group of mice which were fed with chow diet and in group of mice which were fed with low carbohydrate high protein diet. Methods: For this study were used apoE/LDLR deficient female mice which were separated into two groups (in each group were 6 mice). The first group was fed with chow diet (control group) and the second one was fed with low carbohydrate high protein diet (LCHP). This diet was administered to the mice for 8 weeks. After that the imunohistochemical analysis was performed on the cuts of brachiocephalic artery which uses Immpress method with diaminobenzidin (DAB) for better visibility of macrophages. Afterwards was performed biochemical and stereological analysis. Results: Biochemical analysis showed increased levels of total cholesterol in mice with LCHP diet. Immunohistochemical analysis showed presence of macrophages in...
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Immunohistochemical detection of eNOS in atherosclerotic lesions brachiocephalic artery
Klimešová, Kateřina ; Nachtigal, Petr (advisor) ; Němečková, Ivana (referee)
Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Kateřina Klimešová Supervisor: Doc. PharmDr. Petr Nachtigal, Ph.D. Title of diploma thesis: Immunohistochemical detection of eNOS in atherosclerotic lesions brachiocephalic artery Background: We observed the expression of endothelial NO synthase (eNOS) in brachiocephalic artery in apoE/ LDLR-/- deficient mice fed by low carbohydrate high protein diet. Methods: We used apoliporotein E and LDL- receptor deficient female mice that are prone to atherosclerosis. Mice were divided in two groups, 6 mice in each group. First group (control) was fed by western-type diet; second group was fed by low carbohydrate high protein diet (LCHP). We made total cholesterol level measurement. The expression of eNOS was detected by imunohistochemical and stereological methods. Results: Biochemical analysis showed significantly increased total cholesterol level in LCHP group comparing to control group. eNOS expression was detected by imunohistochemical analysis in endothelium covering atherosclerotic plagues as well as in intact endothelium in both groups. Stereological analysis has detected no significant increase of eNOS expression in LCHP group comparing to control group. Conclusion: Increased total...
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