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Study of the variations in the expression of different adhesion and cytoskeletal proteins of podocytes (E-Cadherin, Podocin, Vimentin) due to Bisphenol A
Chvojanová, Zuzana ; Kovařík, Miroslav (advisor) ; Němečková, Ivana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové, Department of Biological and Medical Sciences The University of Alcalá, Faculty of Medicine, Department of biomedicine and biotechnology Student: Zuzana Chvojanová Supervisor: PharmDr. Miroslav Kovařík, Ph.D. Consultant: María Isabel Arenas Jimenéz Title of the diploma thesis: Study of the variations in the expression of different adhesion and cytoskeletal proteins of podocytes (E-Cadherin, Podocin, Vimentin) due to Bisphenol A Bisphenol A (BPA) is one of the most widespread compounds in the world, producing over 6 billion metric tons per year. It is widely used as part of polycarbonate plastics and epoxy resins, from which reusable plastic bottles, food boxes and some medical equipment are made. It is also used to coat the inner layer of the cans. Previous studies have shown that BPA contributes to many chronic diseases in the human body, such as kidney disease - diabetic nephropathy. Podocytes - terminally differentiated cells of the Bowman's capsule in glomerulus - are an integral part of the filtration barrier, where they play an important role in preventing the plasmatic proteins from penetrating to the urine. Therefore, in this study, we looked at the effect of BPA on these cells and their particular proteins, using both in vivo and...
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Study of the variations in the expression of different adhesion and cytoskeletal proteins of podocytes (E-Cadherin, Podocin, Vimentin) due to Bisphenol A
Chvojanová, Zuzana ; Kovařík, Miroslav (advisor) ; Němečková, Ivana (referee)
Charles University, Faculty of Pharmacy in Hradec Králové, Department of Biological and Medical Sciences The University of Alcalá, Faculty of Medicine, Department of biomedicine and biotechnology Student: Zuzana Chvojanová Supervisor: PharmDr. Miroslav Kovařík, Ph.D. Consultant: María Isabel Arenas Jimenéz Title of the diploma thesis: Study of the variations in the expression of different adhesion and cytoskeletal proteins of podocytes (E-Cadherin, Podocin, Vimentin) due to Bisphenol A Bisphenol A (BPA) is one of the most widespread compounds in the world, producing over 6 billion metric tons per year. It is widely used as part of polycarbonate plastics and epoxy resins, from which reusable plastic bottles, food boxes and some medical equipment are made. It is also used to coat the inner layer of the cans. Previous studies have shown that BPA contributes to many chronic diseases in the human body, such as kidney disease - diabetic nephropathy. Podocytes - terminally differentiated cells of the Bowman's capsule in glomerulus - are an integral part of the filtration barrier, where they play an important role in preventing the plasmatic proteins from penetrating to the urine. Therefore, in this study, we looked at the effect of BPA on these cells and their particular proteins, using both in vivo and...
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Nutrition of dialysis patient
BOHÁČOVÁ, Kristýna
Bachelor thesis "The Diet of Dialyzed Patients" deals with the nutrition of patients, who regularly receive the dialysis treatment. The aim of this work is to map the awareness of a diet regime and patients' compliance with it. The theoretical part describes the anatomy and physiology of kidneys, then it mentions the kidney failure and the elimination methods. Most of the space in this part is given to the nutrition, where the basic nutrients, vitamins and minerals are described. Their amount is specific for the dialyzed patients. The end of this part is aimed at suitable and unsuitable foodstuffs for dialyzed patients. The practical part uses the quantitative method of structured questionnaire, which contains 34 questions. 30 respondents, who regularly receive the dialysis treatment, participated in it. The results of the research showed that 17 out of 30 patients adhere to the diet measures for dialyzed patients. 57% is not a satisfying result given the availability of the education nowadays. The reason for this result might be that the respondents were not informed about the diet regime from a nutritional therapist. The work suggests the presence of a nutritional therapist for regular check-ups and so-called big blood samples that are done once a month to check the amount of minerals and other substances. Nutritional therapist could then decide, which substances are abundant or deficient, and they could then suggest an adjustment of the patient's diet.
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New diets for patients on dialysis
Kubíková, Kristýna ; Matějková, Miroslava (advisor) ; Zakiyanov, Oskar (referee)
The bachelor thesis focuses on new diets for dialysis patients. The aim is to create a new dietary regime system that meets the dietary requirements during dialysis treatment and also make the work of nutritional therapists easier. The old system was based on "common rational diet no. 3" and the food was selected individualy for every patient in order to avoid unsuitable meals in the dietary regime. This system was rather time consuming and the nutritional values did not meet the requirements of a dialysis diet. The anatomy and physiology emphasis is placed especially on chronic renal insufficiency. The privotal point of this chapter is the dialysis treatment and a description of dietotherapy in this stage of kidney failure. The practical part looks as the methodology of creating a new dietary regime. The result part compares the improvement in nutritional values with the original "rational diet no.3" and also looks at the methodology of creating new dietary regimes. The main research goal of the questionnare was to find out the patient's satisfaction with the new dialysis diet. Another goal was to discover the nutritional balance of individual dishes and the willingness to continue the diet after hospitalization. Finally, I would also like to mention that theoretically prepared dietary regime was put...
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Generation and Characterization of Glutamate Carboxypeptidase II (GCPII)-Deficient Mice
Vorlová, Barbora ; Šácha, Pavel (advisor) ; Eckschlager, Tomáš (referee) ; Bařinka, Cyril (referee)
Glutamate carboxypeptidase II (GCPII) is a transmembrane glycoprotein, which consists of short intracellular and transmembrane domains, and a large extracellular domain possessing carboxypeptidase activity. In the human body, GCPII fulfils a neuromodulatory function in the brain and facilitates folate absorption in the small intestine. In addition to the brain and small intestine, high level of GCPII is also present in the prostate and kidney. However, GCPII function in these tissues has not been determined yet. To study the role of GCPII in detail, several research groups attempted to inactivate GCPII encoding gene Folh1 in mice. Surprisingly, the experiments led to rather conflicting results ranging from embryonic lethality to generation of viable GCPII-deficient mice without any obvious phenotype. This dissertation project aimed to dissect the discrepancy using alternative strategy for gene modification. For this purpose, we designed TALENs that specifically targeted exon 11 of Folh1 gene and manipulated mouse zygotes of C57BL/6NCrl genetic background. We analysed all genetically modified mice of F0 generation for presence of TALEN-mediated mutations and established 5 different GCPII-mutant mouse colonies from founder mice that altogether carried 2 frame-shift mutations and 3 small in-frame...
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The activity ratio of 30. minute/2.-3. minute in the Dynamic renal scintigraphy
KUKAČKOVÁ, Jana
The objective of the present Bachelor thesis is to determine normal values of the ratio of activity in the Region of Interest (ROI) of a healthy kidney in the 30th minute of the examination compared to the 2nd - 3 rd minutes in patients up to 18 years of age at the Nuclear Medicine Department in České Budějovice. For the purposes of comparison, values of the ratio of activity in ROI of the kidney in the 30th minute are compared with the peak in the same patients. Dynamic kidney scintigraphy with 99mTc-MAG3, generally modified by a furosemide test, is carried out in case of child patients with respect to whom simple dilatation of the pelvicalyceal system must be distinguished from obstruction. A set of 35 patients at the age of 0-18 was selected for the purposes of statistical processing in the period from 01 January 2018 to 13 November 2018, whereupon these patients underwent dynamic kidney scintigraphy with 99mTc-MAG3. As a result, 47 usable renal units were obtained. Renal units with pathological findings were removed from the set. In these renal units, the ratio of activity 30th min./3rd min. and the ratio 30th min./peak were observed. For comparison, the total of 4 selective statistical sets (VSS) were formed - a group of patients at the age of 0-18 and a more restricted group of patients at the age of 0-10 years for both ratios of activity. The collected data were subsequently used for parametric testing. The contribution of this thesis consists in the determination of normal values of the ratio of activity 30th minute/2nd - 3 rd minutes, which will be useful in differentiating mild dilatation from other pathological conditions (obstruction, reductions in function, etc.) in patients examined by dynamic kidney scintigraphy at the Nuclear Medicine Department in České Budějovice. The elementary static processing implies that normal values of the ratio of activity 30th min./3rd min. oscillate in the interval of values (0.058; 0.238) in patients up to 18 years of age, and in the interval (0,05; 0.27) in patients up to 10 years of age. For the sake of comparison, values of the ratio of activity 30th min./peak oscillate in the interval of values (0,026; 0.246) in patients up to 18 years of age, and in the interval (0.04; 0.26) in case of patients up to 10 years of age. It may be stated that values of ratios of activity 30th min./3rd min. and 30th min./peak are approximately comparable. Based on the applied double-selection t-test, the zero hypothesis was adopted, meaning that the selective statistical sets originate from the same basic statistical set on the given significance level.
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Differentiation of human M2 monocytes/macrophages and their role in kidney transplantation
Čápová, Barbora ; Stříž, Ilja (advisor) ; Krulová, Magdaléna (referee)
The system of mononuclear phagocytes includes macrophages that modulate their phenotype based on microenvironmental signals. Their properties vary considerably in a differentiated stage. M1 macrophages, which are classically activated (typically by IFN-γ), are involved in phagocytosis and produce some pro-inflammatory cytokines, that can stimulate other immune cells. A phenotypically different cell population are M2 macrophages, which are alternatively activated by exposure by Th2 cytokines. M2 macrophages produce preferentially anti-inflammatory cytokines IL-10, TGF-β and participate in repair and tissue healing. The main aim of this study was to standardize a model of differentiation of THP-1 cells and human monocytes towards the M2 phenotyp using an in vitro model. This is represented in particular by increased expression of CD163 and CD206 molecules. The second aim was to assess the dynamics of expression (and co-expression) of CD163 and CD206 molecules in monocytes of patients after kidney transplantation. Expression of surface markers was determined by flow cytometry. Both THP-1 cells and human monocytes, isolated from buffy coat fraction, were stimulated by IL-4, TNF-α, TGF-β and IL-10. Changes in CD163 and CD206 expression were measured after day 1, day 3 and day 6 of stimulation. The most...
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The nutrition of patients undergoing dialysis
MARTÍNKOVÁ, Barbora
The bachelor thesis "The nutrition of patients undergoing dialysis"(whose count is around 6000 in the Czech Republic), deals with the nutrition of patients who regularly undergo treatment by dialysis. The theoretical part focuses on the anatomy and physiology of kidneys. Moreover, the stages of kidney failure and the possibilities of compensation of the kidney function are discussed. Furthermore, the theoretical part of the thesis focuses on dietary measures by patients undergoing dialysis, suitable and unsuitable groceries and their technological adjustment. The aim of the thesis is to map out the awareness of the patients and their families about the diet of a patient undergoing dialysis and to evaluate the menus of the patients, mainly the presence of basic nutrients potassium, phosphor and calcium in their diets. The practical part of the thesis was carried out by qualitative research. Six respondents who are regularly treated by dialysis participated in the research. On the basis of the evaluation of weekly menu of each respondent and counting guideline daily amounts the result was very negative. All respondents have received less amount of energy then what was counted for them. The same applied for basic nutrients. A positive finding was, that all of the respondents have been successful in reducing potassium and most of them have been able to reduce phosphor. Only one respondent fitted in the recommended amount of calcium for patients undergoing dialysis. From the results is obvious, that only one respondent abides by dietary recommendations. Most of the patients couldn't say which groceries are suitable for them and which are not. Unfortunately, the cause of this is, that not one of the respondents listed a nutrition therapist as a source of dietary measures.
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Morphological characteristics of alterations in lungs in a mouse model of atherosclerosis.
Vyskočilová, Martina ; Němečková, Ivana (advisor) ; Blažíčková, Kateřina (referee)
Bachelor thesis Morphological characteristics of alterations in lungs in a mouse model of atherosclerosis Author: Martina Vyskočilová Study course: Medical laboratory technician Charles University Faculty of Pharmacy in Hradec Králové Background: Atherosclerosis is a multifactorial disease caused by deposition of blood lipids into the intima layer of blood vessels. Due to the fact that process of atherogenesis is associated with immune reactions, disease is considered as an inflammatory process. Atherosclerotic plaques rarely affect pulmonary arteries, where this condition is accompanied by an increase of pulmonary pressure, which is very likely the trigger of disease. Aim of this study is to find out whether there are morphological changes in the lungs of ApoE/LDLr-/- mouse model of atherosclerosis, characteristic for pulmonary atherosclerosis, compared to lungs of healthy mice. Methods: Lungs of ApoE/LDLr-/- and healthy C57BL/6J control mice have been fixed in 4% neutral formaldehyde, embedded in paraffin and sliced into 4 μm thin sections. One of the sections has been stained with hematoxylin-eosin, the other one with green trichrome. Staining with hematoxylin-eosin is primary staining used for basic imaging of histological structure of tissues, staining with green trichrome is used for imaging...
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