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Laboratory diagnosis of microRNA in different diagnostic groups
Hrachovcová, Eliška ; Urbánková Rathouská, Jana (advisor) ; Tripská, Katarína (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Student: Bc. Eliška Hrachovcová Supervisor: PharmDr. Jana Rathouská, Ph.D. Consultant: doc. RNDr. Pavlína Kušnierová, Ph.D. Title of diploma thesis: Laboratory diagnostics of microRNAs in different diagnostic groups Background: Identification of potential miRNA targets for differential diagnosis of patients with atherosclerosis and dementia. Comparison of miRNA expression and uptake in serum samples with cerebrospinal fluid samples. Selection of appropriate miRNAs for differential diagnosis in patients with atherosclerosis and dementia. Furthermore, the selection of miRNA targets with the most significant fold-change and also the lowest p-value for subsequent validation studiues. Methods: To identify potential miRNAs, the received samples were centrifuged, aliquoted and stored at - 80C. miRNAs were isolated using the iCatcher Circulating cfRNA 1000 kit and the CatchGene isolator. Reverse transcription and amplification with detection were then performed. Data were processed in Excel and then statistically processed in GenEx using Mann-Whitney test. Results: The results were evaluated for 2 groups of patients: group A (samples of patients with Parkinson's disease and dementia) and group B (with...
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Effect of atorvastatin on endoglin expression and function in hyperlipidemia-induced endothelial dysfunction
Holadová, Adéla ; Tripská, Katarína (advisor) ; Fikrová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Effect of atorvastatin on endoglin expression and function in hyperlipidemia-induced endothelial dysfunction Author: Adéla Holadová Supervisor of Diploma Thesis: PharmDr. Katarína Tripská, Ph.D. Aim: The aim of this thesis was to determine how atorvastatin affects the expression and function of endoglin and related biomarkers of inflammation/endothelial dysfunction at different concentrations and times in human endothelial cells from the aorta, in which hyperlipidemia was induced by using palmitic acid. Methods: In the diploma thesis, we worked with human aortic endothelial cells. We premedicated them with atorvastatin/palmitic acid at different times and concentrations. Using flow cytometry, we measured the protein expression of endoglin. Furthermore, we used the qRT-PCR method to determine the level of mRNA expression of endoglin, its transcription factors and biomarkers of inflammation. Using the ELISA method, we detected the levels of soluble endoglin. Results: We demonstrated that atorvastatin at a concentration of 5μM after 16 hours of premedication significantly increased the mRNA expression of endothelial nitric oxide synthase, and conversely decreased the...
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Effect of glipizide on the expression and function of endoglin and related biomarkers of endothelial dysfunction in diabetes mellitus type II
Vochozková, Iva ; Tripská, Katarína (advisor) ; Urbánková Rathouská, Jana (referee)
8 ABSTRACT Charles University, Faculty of Pharmacy in Hradec Kralové Department of Biological and Medical Sciences Title of Diploma Thesis: Effect of glipizide on the expression and function of endoglin and related biomarkers of endothelial dysfunction in type II diabetes mellitus Author: Iva Vochozková Supervisor of Diploma Thesis: PharmDr. Katarína Tripská, Ph.D. Aim: The aim of this thesis was to determine how glipizide affects the expression and function of endoglin, its transcription factors and related biomarkers of endothelial dysfunction, soluble endoglin levels and monocyte adhesion to human diabetic coronary artery endothelial cells. Methods: In this thesis we worked with human diabetic coronary artery endothelial cells. These were premedicated with glipizide at various times and concentrations. We measured protein expression of endoglin and biomarkers of endothelial dysfunction using flow cytometry. We also measured monocyte adhesion to endothelial cells using flow cytometry. We detected soluble endoglin levels by ELISA and measured mRNA expression of endoglin, its transcription factors and markers of inflammation by PCR. Results: We demonstrated that premedication with 200µM concentration of glipizide led to a decrease in endoglin protein expression after 24 and 48 hours. The protein expression...
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Effect of endoglin blockage on inflammation-induced endothelial dysfunction in human aortic endothelial cells
Škubová, Andrea ; Tripská, Katarína (advisor) ; Svobodová, Zuzana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Effect of endoglin blockage on inflammation-induced endothelial dysfunction in human aortic endothelial cells Author: Andrea Škubová Supervisor of Diploma Thesis: PharmDr. Katarína Tripská, Ph.D. Aim: The aim of this thesis was to describe the effect of TNF-α and LPS-induced inflammation on the expression of endoglin and its transcription factors, the expression of biomarkers of endothelial dysfunction, the levels of soluble endoglin, and the adhesion and transmigration of monocytes through human aortic endothelial cells. We further investigated the effect of endoglin blockage by the monoclonal antibody TRC105 (carotuximab) on these processes. Methods: In this thesis, human aortic endothelial cells (HAEC) were used, and they were exposed for 16 hours to TNF-α (10 ng/ml) and LPS (100 ng/ml) and for 12 hours to TRC105 (300 µg/ml). The mRNA expression of endoglin and its transcription factors (KLF6, RelA, NR1H3), mRNA expression of inflammatory adhesion molecules (ICAM-1, VCAM-1, SELE) and mRNA expression of MMP-14 enzyme were measured by real time qRT-PCR. Using flow cytometry, we detected protein levels of endoglin, protein levels of inflammatory adhesion molecules...
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Effect of endoglin blockage on inflammation-induced endothelial dysfunction in human aortic endothelial cells
Škubová, Andrea ; Tripská, Katarína (advisor) ; Svobodová, Zuzana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Effect of endoglin blockage on inflammation-induced endothelial dysfunction in human aortic endothelial cells Author: Andrea Škubová Supervisor of Diploma Thesis: PharmDr. Katarína Tripská, Ph.D. Aim: The aim of this thesis was to describe the effect of TNF-α and LPS-induced inflammation on the expression of endoglin and its transcription factors, the expression of biomarkers of endothelial dysfunction, the levels of soluble endoglin, and the adhesion and transmigration of monocytes through human aortic endothelial cells. We further investigated the effect of endoglin blockage by the monoclonal antibody TRC105 (carotuximab) on these processes. Methods: In this thesis, human aortic endothelial cells (HAEC) were used, and they were exposed for 16 hours to TNF-α (10 ng/ml) and LPS (100 ng/ml) and for 12 hours to TRC105 (300 µg/ml). The mRNA expression of endoglin and its transcription factors (KLF6, RelA, NR1H3), mRNA expression of inflammatory adhesion molecules (ICAM-1, VCAM-1, SELE) and mRNA expression of MMP-14 enzyme were measured by real time qRT-PCR. Using flow cytometry, we detected protein levels of endoglin, protein levels of inflammatory adhesion molecules...
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Current trends in dyslipidemia therapy
Krupová, Tatiana ; Nachtigal, Petr (advisor) ; Tripská, Katarína (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Current trends in dyslipidemia therapy Author: Tatiana Krupová Supervisor of Diploma Thesis: prof. PharmDr. Petr Nachtigal, Ph.D Background: The purpose of this diploma thesis is to describe therapeutic options of different types of dyslipidemia with focus on current trends and new discoveries regarding this matter. Main findings: This diploma thesis emphasizes the importance of correct treatment of dyslipidemia using well-known drugs, as well as new groups of medications to enable the prevention of cardiovascular diseases, the main cause of mortality worldwide. Conclusion: The information gathered for this diploma thesis confirms significant progress in the development of new types of treatment, which consequently makes it possible to better prevent complications that are mainly manifested by the development of cardiovascular diseases. Keywords: cholesterol, lipids, lipoproteins, dyslipidemia, coronary heart disease, statins, fibrates, sequestrants, ezetimibe, niacin, omega-3 fatty acids, phytosterols, fiber, PCSK9 inhibitors, bempedoic acid, mipomersen, volanesorsen, lomitapide, evinacumab, inclisiran, LXR agonists
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Effect of dapagliflozin on the expression and function of endoglin and related biomarkers of endothelial dysfunction in human diabetic coronary endothelial cells
Vavreková, Valéria ; Tripská, Katarína (advisor) ; Fikrová, Petra (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Title of Diploma Thesis: Effect of dapagliflozin on the expression and function of endoglin and related biomarkers of endothelial dysfunction in human diabetic coronary artery endothelial cells Author: Valéria Vavreková Supervisor of Diploma Thesis: PharmDr. Katarína Tripská, Ph.D. Aim: The aim of this thesis was to investigate how dapagliflozin affects the expression and function of endoglin (Eng) and related biomarkers of endothelial dysfunction at different times and concentrations in human diabetic cells. Methods: In this thesis, we worked with human diabetic coronary endothelial cells. We premedicated these with dapagliflozin at different times and concentrations, and dimethyl sulfoxide was used as a control. We measured protein expression of Eng and biomarkers of endothelial dysfunction using flow cytometry. We also used flow cytometry to measure the rate of monocyte adhesion to endothelial. Using ELISA, we detected soluble endoglin (sEng) levels. Results: We demonstrated that a 1nM concentration of dapagliflozin during a 12-hour premedication period led to a decrease in protein expression of adhesion molecules. We further showed that protein expression of Eng was also significantly reduced...
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Possible changes of endoglin expression in endothelial cells.
Tripská, Katarína ; Nachtigal, Petr (advisor) ; Mladěnka, Přemysl (referee) ; Vohnout, Branislav (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Doctoral Degree Program: Pharmacology and Toxicology Candidate: Mgr. Katarína Tripská Supervisor: prof. PharmDr. Petr Nachtigal, Ph.D. Title of Doctoral Thesis: Possible changes of endoglin expression in endothelial cells Endoglin (Eng) is a co-receptor for ligands of the Transforming Growth Factor β superfamily. Multiple studies have demonstrated that Eng is associated with metabolic syndrome-related pathological conditions, such as hypercholesterolemia, hyperglycemia, hypertension, or obesity. However, the exact role of Eng in these pathologies is still unknown. Therefore, the main purpose of this thesis was to elucidate and summarize the role of membrane and soluble Eng in the selected pathologies associated with metabolic syndrome. We demonstrated that soluble Eng (sEng) levels, together with plasma lipids and markers of inflammation, were reduced after every lipoprotein apheresis in patients with familial hypercholesterolemia. This result, in combination with knowledge acquired by the review article, indicates that increased sEng levels are associated with cardiometabolic disorders. Regarding the role of membrane Eng, we showed that blockage of Eng expression by TRC105 results in a decrease...
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Possible changes of endoglin expression in endothelial cells.
Tripská, Katarína ; Nachtigal, Petr (advisor) ; Mladěnka, Přemysl (referee) ; Vohnout, Branislav (referee)
Charles University, Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Doctoral Degree Program: Pharmacology and Toxicology Candidate: Mgr. Katarína Tripská Supervisor: prof. PharmDr. Petr Nachtigal, Ph.D. Title of Doctoral Thesis: Possible changes of endoglin expression in endothelial cells Endoglin (Eng) is a co-receptor for ligands of the Transforming Growth Factor β superfamily. Multiple studies have demonstrated that Eng is associated with metabolic syndrome-related pathological conditions, such as hypercholesterolemia, hyperglycemia, hypertension, or obesity. However, the exact role of Eng in these pathologies is still unknown. Therefore, the main purpose of this thesis was to elucidate and summarize the role of membrane and soluble Eng in the selected pathologies associated with metabolic syndrome. We demonstrated that soluble Eng (sEng) levels, together with plasma lipids and markers of inflammation, were reduced after every lipoprotein apheresis in patients with familial hypercholesterolemia. This result, in combination with knowledge acquired by the review article, indicates that increased sEng levels are associated with cardiometabolic disorders. Regarding the role of membrane Eng, we showed that blockage of Eng expression by TRC105 results in a decrease...
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Evaluation of antibody quality for Endoglin detection in liver
Kejzlarová, Eliška ; Svobodová, Zuzana (advisor) ; Tripská, Katarína (referee)
The diploma thesis deals with endoglin, a transmembrane glycoprotein that can be cleaved to form so-called soluble endoglin, which then circulates in the blood. The theoretical part of the thesis is devoted to cardiovascular and liver diseases related to changes in the expression or level of endoglin in the blood and summarizes current knowledge about endoglin and its relation to various diseases. The experimental part of the thesis describes the selection of a suitable antibody against endoglin intended for western blot (WB). We have four anti-endoglin primary antibodies available from three manufacturers. The purpose was to test the antibodies and determine, which one had the best ability to detect endoglin in mouse tissue samples such as liver or aorta on WB. Affiblot was used to verify their ability to bind endoglin and also to compare their affinities to this protein. Newly developed patented dot-blot-based device. The binding of the used antibodies to endoglin was verified using the WB method and then the results of both methods were compared. The obtained results showed that the Abcam antibody has the most suitable parameters regarding specificity and sensitivity for the identification of endoglin in mouse tissue on WB. Keywords: endoglin, soluble endoglin, NASH, cirrhosis, non-alcoholic...
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