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Changes of endothelial expression of endoglin during atherogenesis
Zajícová, Šárka ; Nachtigal, Petr (advisor) ; Rathouská, Jana (referee)
Šárka Zajícová Changes of endothelial expression of endoglin during atherogenesis Diploma thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Specialist in Laboratory Methods Background: We observed the expression of endoglin in arteria brachiocephalica in the apoE−/− /LDLr−/− deficient mice after feeding them with the low carbohydrate, high protein diet. Methods: We used the female apoE−/− /LDLr−/− deficient mice, which we divided into two groups after 6 animals. One of these groups were fed by AIN-93G diet (control group) and the second one by low carbohydrate, high protein diet. Stereological and immunohistochemical analysis was performed in tissue sections of arteria brachiocephalica by means of ImmPRESSTM method with DAB detection. Results: Biochemical analysis showed a significant increase levels of total cholesterol after feeding with low carbohydrate, high protein diet. Immunohistochemical analysis, we demonstrated endoglin expression in endothleial cells, mostly covering atherosclerotic plaques in both groups apoE−/− /LDLr−/− deficient mice. Stereological analysis in mice fed with low carbohydrate, high protein diet showed weak endoglin expression when compared to group on chow diet. Conclusions: Mice fed with low carbohydrate, high protein diet showed increased...
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Markers of inflammation and proliferation in the heart of a transgenic mouse model expressing high levels of soluble endoglin
Boukalová, Petra ; Rathouská, Jana (advisor) ; Fikrová, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Markers of inflammation and proliferation in the heart of a transgenic mouse model expressing high levels of soluble endoglin Rigorous thesis Mgr. Petra Boukalová Consultant of rigorous thesis: PharmDr. Jana Rathouská, Ph.D. Background: The aim of this thesis was to investigate the expression of markers of inflammation and proliferation in the cardiac wall of genetically modified mice expressing high levels of soluble endoglin. It was shown that soluble endoglin molecule plays a role in many cardiovascular diseases. The transgenic mouse model with high levels of soluble endoglin allows us to study its contribution to the pathogenesis of these diseases. Methods: For this work a transgenic mouse model on CBAxC57BL/6J backgroud with high levels of human soluble endoglin was used. We observed the expression of selected molecules (ICAM-1, VEGF, COX-2 and SOD-3) in the heart wall of this group of mice compared to the control group of mice with low levels of human soluble endoglin. Both groups of animals were fed a chow diet till the age of nine months. Western blot analysis was used to detect the protein expression. ELISA analysis was used to allocate the mice in two relevant groups. Results:...
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Western blot analysis of selected oxidative stress markers in aorta of apoE/LDLR deficient mice
Čermáková, Martina ; Rathouská, Jana (advisor) ; Blažíčková, Kateřina (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Western blot analysis of selected oxidative stress markers in aorta of apoE/LDLR deficient mice Author of diploma thesis: Bc. Martina Čermáková Supervisor: PharmDr. Jana Rathouská, Ph.D. Background: The aim of this diploma thesis was to evaluate expression of selected oxidative stress molecules (SOD-3, iNOS, catalase, HO-1) in cardiac aorta and nitrate and nitrite levels in plasma in two groups of mice. These two groups of mice varied in genetic features - the monitored group was, compared to control group of mice, deficient for apolipoprotein E and LDL receptor. Methods: For the study, 2 months old male mice were fed a standard rodent diet. The males were divided into two groups - control (C57BL/6J) and monitored (apoE-/- /LDLR- /- ). Western blot method was used to evaluate and quantify iNOS, SOD-3, catalase and HO-1 molecules in the aorta. HPLC method was used to quantify nitrate and nitrite levels in plasma. Result: HPLC method did not show statistically significant differences in the nitrate and nitrite levels in plasma between the two groups of examined mice. Western blot analysis of aortic samples showed statistically significantly higher SOD-3 expression (208.39 %) and a significantly...
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Evaluation of endoglin and VCAM-1 expression and co-expression in aortas of apoE-deficient mice
Minaříková, Lucie ; Rathouská, Jana (advisor) ; Fikrová, Petra (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Evaluatiton of the expresion and co-expresion of endoglin and VCAM-1 in the aorta on apoE- deficient mice Diploma thesis Lucie Minaříková Supervisor: Mgr. Jana Rathouská Background: The aim of this work was to monitor the expression and a possible co-expression of endoglin (TGF-β receptor III) and VCAM-1 in mouse aortic endothelium. As an experimental model, we used the mouse strain C57BL/6J, that was genetically modified, and is characteristic by a deficit of apolipoprotein E. Methodes: In the study, we focused on testing the mouse strain C57BL/6J with gene knockout for apolipoprotein E in different stages of the atherosclerotic process. 10 weeks old female mice were divided into three groups and fed diets with different content of cholesterol. One experimental group was fed a standard diet (called "chow type diet") for a period of two months. The other two groups were fed a diet containing 21% fat (called "Western type diet") for a period of two and four months. For the determination of the levels of total cholesterol, a biochemical analysis of blood was performed. Obtained parts of ascending aorta were analyzed by ImmPRESSTM immunohistochemical method with the detection reagent DAB....
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Immunohistochemical detection of PECAM-1, endoglin and VCAM-1 in aorta of transgenic mice with high levels of soluble endoglin
Brlicová, Monika ; Rathouská, Jana (advisor) ; Filipský, Tomáš (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Immunohistochemical detection of PECAM-1, endoglin and VCAM-1 in aorta of transgenic mice with high levels of soluble endoglin Rigorous thesis Mgr. Monika Brlicová Consultant of rigorous thesis: PharmDr. Jana Rathouská, Ph.D. Background: We observed an effect of high level of human soluble endoglin (sEng) on an expression of tissue endoglin and adhesion molecules VCAM-1 and PECAM-1 in descendent aortas of experimental animals. Methods: 6 months old female transgenic mice of strain CBAxC5B7BL/6J with high plasma levels of human soluble endoglin (Sol-Eng+ ) and the same types of mice, which show signs of low plasma levels of human soluble endoglin as control mice were used for this rigorous thesis. Both groups were fed by high fat diet for 3 months. After that the levels of total cholesterol in the blood of these animals were biochemically determined and then statistically evaluated. The immunohistochemical method ImmPRESSTM was used for detection of protein expression in aortic sections. Results: Biochemical analysis showed a statistically insignificant difference in total cholesterol levels in both groups. Immunohistochemical analysis ImmPRESSTM showed no differences in the expression of...
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Evaluation of endoglin and P-selectin expression and co-expression in aortas of apoE-deficient mice
Brlicová, Monika ; Rathouská, Jana (advisor) ; Kovařík, Miroslav (referee)
Charles University in Prague Faculty of Pharmacy in HradecKrlov Department of Biological and Medical Sciences Evaluation of endoglin and P-selectin expression and co-expression in aortas of apoE-deficient mice Diploma thesis MonikaBrlicov Supervisor: Mgr.JanaRathousk Background: We observed the expression and the reciprocal co-expression of endoglin (receptor III for TGF- cytokine) and P-selectin (adhesion molecule and marker of endothelial dysfunction) in ascending aortas of apoE-deficient mice which were fed by standard diet for rodents and Western type diet (high-cholesterol diet) for achieving of different phases of the atherosclerotic process. The changes of total cholesterol levels in mice after administration of both types of diets were also evaluated. Methods: The modified strain C57BL/6J of mice with a deficiency of apolipoprotein E, which is prone to aterogenesis was used for this diploma thesis. Mice were divided into three groups. The first group was fed by standard diet (so-called "chow" diet) for a period of two months and the second two groups were fed by Western type diet for a period of two and four months. The levels of total cholesterol in the blood were biochemically determinated and then we statistically evaluated this levels in all groups. Immunohistochemical...
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Monitoring of expression changes of selected markers in glucose-induced endothelial dysfunction in vitro.
Hudecová, Alexandra ; Nachtigal, Petr (advisor) ; Rathouská, Jana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Monitoring the change in expression of selected indicators in the glucose induced endothelial dysfunction in vitro. Author: Alexandra Hudecová Supervisor of Diploma Thesis: prof. PharmDr. Petr Nachtigal, Ph.D. Background: The aim of this diploma thesis was to determine, whether the new monoclonal antibody carotuximab affects the expression of endoglin and other biomarkers of endothelial dysfunction (eNOS, ICAM-1, VCAM-1, E-selectin) in human aortic endothelial cells exposed to high glucose. Methods: Human aortic endothelial cells (HAEC) were exposed to high glucose levels (45 mmol) for 72 hours and carotuximab (300 µg/ml) for 12 hours. We measured the gene expression of endoglin, eNOS, ICAM-1, VCAM-1, E-selectin by real-time PCR. We measured endoglin and ICAM-1 protein levels by flow cytometry. Results: Gene expression of endoglin and all biomarkers of endothelial dysfunction was significantly increased after high glucose. After the addition of carotuximab to high glucose, we observed a significant increase in gene expression of eNOS, while no significant difference was measured in endoglin or other markers of endothelial dysfunction. Protein levels of endoglin were...
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Neonatal birth injury and therapy
Fenclová, Eva ; Rathouská, Jana (advisor) ; Nejmanová, Iveta (referee)
Background: The basis for the prevention of birth injuries is the knowledge of their etiopathogenesis and the mechanism of labor. If birth trauma occurs, it is important to take a proper care of the injured newborn after birth. The aim of this work is to create a set of information about birth injuries, their causes and possible therapy. Main findings: Birth injuries arise as a result of the risk course of childbirth, but they can also arise as a result of risk factors on the part of the mother or fetus. Most of these traumas do not require any therapy because they heal spontaneously, but some require surgical treatment. Conclusions: Information was sought on birth injuries, childbirth, its mechanism, newborns and its care. The literature search for the topic of birth traumas was focused on the causes, incidence, manifestations, consequences and treatment. Information was obtained from both Czech and foreign sources. Keywords: newborn, labor, birth injury, asphyxia, surgical delivery
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Monitoring of expression changes of selected markers in glucose-induced endothelial dysfunction in vitro.
Hudecová, Alexandra ; Nachtigal, Petr (advisor) ; Rathouská, Jana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Monitoring the change in expression of selected indicators in the glucose induced endothelial dysfunction in vitro. Author: Alexandra Hudecová Supervisor of Diploma Thesis: prof. PharmDr. Petr Nachtigal, Ph.D. Background: The aim of this diploma thesis was to determine, whether the new monoclonal antibody carotuximab affects the expression of endoglin and other biomarkers of endothelial dysfunction (eNOS, ICAM-1, VCAM-1, E-selectin) in human aortic endothelial cells exposed to high glucose. Methods: Human aortic endothelial cells (HAEC) were exposed to high glucose levels (45 mmol) for 72 hours and carotuximab (300 µg/ml) for 12 hours. We measured the gene expression of endoglin, eNOS, ICAM-1, VCAM-1, E-selectin by real-time PCR. We measured endoglin and ICAM-1 protein levels by flow cytometry. Results: Gene expression of endoglin and all biomarkers of endothelial dysfunction was significantly increased after high glucose. After the addition of carotuximab to high glucose, we observed a significant increase in gene expression of eNOS, while no significant difference was measured in endoglin or other markers of endothelial dysfunction. Protein levels of endoglin were...
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Quality of life evaluation in pregnant women in the Czech republic and Slovakia by means of a specific questionaire QOL-GRAV
Bujňáková, Terézia ; Rathouská, Jana (advisor) ; Hronek, Miloslav (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of diploma thesis: Quality of life evaluation in pregnant women in the Czech republic and Slovakia by means of a specific questionaire QOL-GRAV Author: Terézia Bujňáková Supervisor: PharmDr. Jana Rathouská, Ph.D. Backround First, an overview of the theory concerning quality of life, pregnancy and folic acid intake was the part of the main aims of this thesis. Experimental section is focused on evaluating quality of life of pregnant women, supplementation of folic acid before and during gravidity and its potential effect on quality of life of women in the Czech republic and Slovakia, evaluated by a specific questionaire QOL-GRAV. Methods We used a quntitative method in the form of specific questionaire QOL-GRAV, in which questions about folic acid supplementation and eating habits changes before and during pregnancy, were implemented. 564 pregnant women from Czech and Slovak republic were questioned in any week of gravidity or two weeks after delivery, at latest. Results We can consider quality of life of pregnant women in Czech and Slovak republic as very good, according to average value 18,81 points of 45 points overall obtained from the questionaire, where minimum possible value of 9...
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