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Characterization of blackthorn products
Červinková, Zuzana ; Veselá, Mária (referee) ; Kovalčík, Adriána (advisor)
This bachelor thesis deals with the preparation and the chemical characterization of ethanolic extracts and products from the blackthorn (Prunus spinosa L.) fruit. The theoretical part of the work contains the characterization of the blackthorn and describes the chemical composition of the fruit. Furthermore, the theoretical part states possible effects and possible uses of the Prunus spinosa L. fruit in the food industry, folk medicine and pharmacy. The experimental part describes the preparation of the extracts (40% ethanol) and the products (i.e., homemade sloe wine and homemade blackthorn liqueur). The content of reducing sugars, polyphenols and flavonoids was quantified in samples of wine, liqueur, juice and ethanol extracts. Furthermore, the antioxidant activity of individual samples was determined. Two spectrophotometric methods for the determination of reducing sugars were used and compared, namely the Somogyi-Nelson (SN) assay and the 3,5-dinitrosalicylic acid (DNS) assay. The DNS assay provided slightly overestimated results, while the results from the SN assay were closer to the actual content of reducing sugars. Polyphenols and flavonoids are substances that have antioxidant properties. A strong correlation was observed between antioxidant activity and the concentration of flavonoids.
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Characterization of Mitochondrial Respiratory Chain Defects Using DNA Microarrays
Čížková, Alena ; Kmoch, Stanislav (advisor) ; Kováč, Ladislav (referee) ; Červinková, Zuzana (referee)
A mitochondrion is a organelle under nuclear and mitochondrial genetic control and its maintence requires a nucleo-mitochnodrial cross-talk. Better understanding of mitochondrial pahologies using various gene expression techniques. This thesis presents the implementation and application of microarray technology for identification of disease causing genes in respiratory chain disorders, expecially in patients with ATP synthase deficiency of nuclear origin. The introduction to this thesis provides an overview of mitochondrial structure and biogenesis, respiratory chain complexes and their disorders as well as different types of microarrays. A study of mitochondrial ATP synthase deficiency represents the major experimental part of this thesis and is accordingly that were used in further studies centered on characterization of inherited metabolic disorders. This is documented in positional cloning of patientes with lysosomal storage disorders - mucopolysaccharidosis IIIC.
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Recipe of a food supplement based on bioactive substances isolated from Prunus spinosa
Červinková, Zuzana ; Hudečková, Helena (referee) ; Kovalčík, Adriána (advisor)
Blackthorn fruits (Prunus spinosa) are small blue-purple drupes containing a range of bioactive substances that give them diverse biological effects. They are traditionally used not only in folk medicine, as they are attributed with many preventive and therapeutic effects. The presented master´s thesis deals with the preparation and characterization of an ethanol extract and dietary supplements based on bioactive substances isolated from the fruits of blackthorn. The ethanol extract was prepared by maceration fresh Prunus spinosa fruits in 40% alcohol. The extract was analyzed for the polyphenols (2111.39±11.32 mg GAE/l), flavonoids (903.89±5.08 mg CAE/l) and anthocyanins (165.24±0.38 mg CGE/l) content, antioxidant (11.99±0.40 mmol TE/l) and antimicrobial activity, phenolic compounds profile and sugars profile. The ethanolic extract exhibited antibacterial effect against the gram-positive bacterium Micrococcus luteus and gram-negative bacterium Escherichia coli but was not effective against the yeast Candida glabrata. After gentle concentration on a vacuum evaporator, the extract was used to prepare a dietary supplement. Two types of dietary supplements were prepared, namely gelatine candies with blackthorn fruit extract and gelatine capsules filled with lyophilized fruit powder. The nutritional profile of these products, content of bioactive substances (i.e. total polyphenols, flavonoids, and anthocyanins), phenolic compound profile and antioxidant activity were characterized. Additionally, the release of polyphenols was monitored during a simulated in vitro digestion. The gelatine candies with different extract concentrations (5, 10, 15 and 20 %) were subjected to sensory evaluation, where the aim was to optimize the addition of the extract to the gelatine matrix. According to the preferences of the evaluators, the sample with 15 % extract was rated as the best in terms of overall taste. In the ranking test, this sample was ranked second, behind the sample with 5 % extract, which, on the other hand, received a worse sensory profile rating. The thesis also included monitoring the effect of freezing, lyophilization, and drying on the bioactive compounds content (i.e. total polyphenols, flavonoids, anthocyanins) and antioxidant activity of Prunus spinosa fruits. The highest contents of total polyphenols (57.49±0.33 mg GAE/g DW), flavonoids (45.75±0.12 mg CAE/g DW) and anthocyanins (6.41±0.19 mg CGE/g DW) were quantified in lyophilized fruits, but the highest antioxidant activity (281.72±0.70 mol TE/g DW) was observed in frozen fruits. A statistically significant moderately strong positive correlation (0.641) of antioxidant activity and total flavonoids was observed for the analyzed samples. The results of the thesis showed a great potential for the use of blackthorn berries in the nutraceutical industry, where they can contribute as active additives to increasing both the bioactive and nutritional quality of dietary supplements.
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Metabolic and genetic risks of development of non-alcoholic fatty liver disease in patients after liver transplantation and its impact on patient and graft survival
Míková, Irena ; Trunečka, Pavel (advisor) ; Červinková, Zuzana (referee) ; Brůha, Radan (referee)
Nonalcoholic fatty liver disease (NAFLD) occurs frequently not only in the general population, but also in liver transplant (LT) recipients. The data about prevalence, evolution, causes and significance of steatosis in patients after LT are limited. In a large retrospective study in LT recipients with histological evaluation of steatosis, we found high prevalence of steatosis (56,4 %) and steatohepatitis (10,4 %), the prevalence of steatosis increased after LT. Pretransplant predictors of steatosis included alcoholic cirrhosis and high BMI, whereas increased alkaline phosphatase and mycophenolate mofetil given initially were protective. Posttransplant predictors of steatosis included BMI, serum triglycerides, alcohol consumption and presence of type 2 diabetes mellitus, whereas increased serum creatinine was protective. Presence of significant steatosis/steatohepatitis was not associated with increased grade of fibrosis. There was no difference between the occurrence of steatosis in surviving and lost grafts. Survival of patients with/without significant steatosis was similar with a trend to higher long-term mortality of patients with significant steatosis. In the evaluation of the impact of TM6SF2 rs58542926 and PNPLA3 rs738409 genotypes of the donor and recipient on pathophysiology of steatosis...
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The influence of the treatment of experimental acute liver failure by FPSA/Prometheus on intracranial pressure
Pražák, Josef ; Cvachovec, Karel (advisor) ; Červinková, Zuzana (referee) ; Kieslichová, Eva (referee)
Background: Cerebral edema is a well-known and fatal complication of acute liver failure (ALF). The influence of the treatment by artificial liver devices on the cerebral damage caused by ALF is usually monitored by only measuring the intracranial pressure (ICP). The aim of this work was to determine the influence of Fractionated Plasma Separation and Adsorption (FPSA), nonbiological artificial liver device, on the intracranial pressure. The second goal of this work was to determine the potential role of cerebral microdialysis in monitoring of the efficiency of fractionated plasma separation and adsorption (FPSA) treatment of ALF. Methods: Two types of surgical resection model of ALF were used in two separate experiments in pigs. In the first study data from monitoring of the intracranial pressure of ALF group (animals with ALF only) and FPSA group (animals with ALF treated by FPSA) were compared. In the second study data from monitoring by cerebral microdialysis and ICP of the ALF group (animals with ALF only), FPSA group (animals with ALF treated by FPSA) and SHAM group (animals with only laparotomy) were compared using statistical analyses. Results: In both experiments, the ICP was significantly higher in the ALF group than in the FPSA group from the 9th hour of the experiment. In the second...
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Immune Reactivity of Hepatic T Lymphocytes and Their Role in Pathogenesis of Liver Diseases
Carey, Ivana ; Vítek, Libor (advisor) ; Urbánek, Petr (referee) ; Červinková, Zuzana (referee)
The interaction of different pathogens with systemic and liver immune system is suggested an important factor in pathogenesis of various liver diseases. The variable aspects of this interaction and different immune system patways were subject of the present thesis. The ultimate goal of the present thesis was to investigate the reactivity of the local and systemic immune system against different disease related antigens and to demonstrate their participation in liver diseases and liver damage pathogenesis. Chronic hepatitis B infection, chronic hepatitis C infection and drug-induced liver damage were investigeted by 7 different studies using ex-vivo and in-vitro methods. In general, chronic liver diseases are associated with increased oxidative stress. Due to this fact, the role bilirubin, a potent endogenous antioxidant, was also analysed. The results of presented studies demonstrate clearly active participation of immune system to the pathogenesis of liver diseases, its direct involvement in liver damage and its effect on outcome of disease. The protective role of bilirubin in oxidative stress was confirmed. In conclusion, the results of all presented studies in this thesis illustrate the role of systemic as well as liver based immune system as a crucial player in the pathogenesis of liver...
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Functional characterisation of new components of mitochondrial proteome.
Kovalčíková, Jana ; Vrbacký, Marek (advisor) ; Červinková, Zuzana (referee) ; Ješina, Pavel (referee)
1 Abstract It has been estimated that the mammalian mitochondrial proteome consists of ~1500 distinct proteins and approximately one quarter of them is still not fully characterized. One of these proteins is TMEM70, protein involved in the biogenesis of the eukaryotic F1Fo-ATP synthase. TMEM70 mutations cause isolated deficiency of ATP synthase often resulting in a fatal neonatal mitochondrial encephalocardiomyopathies in patients. To understand the molecular mechanism of TMEM70 action, we generated constitutive Tmem70 knockout mice, which led to embryonic lethal phenotype with disturbed ATP synthase biogenesis. Subsequently generated inducible Tmem70 mouse knockout was lethal by the week 8 post induction. It exhibited primarily impaired liver function, which contrasts with the predominantly cardiologic phenotype at disease onset in humans. Liver mitochondria revealed formation of labile ATP synthase subcomplexes lacking subunit c. Thus, in case of TMEM70 deficiency c-oligomer was not incorporated into ATP synthase, which led to critical impairment of mitochondrial energy provision, analogous to TMEM70 dysfunction in humans. In TMEM70 deficient models, the ATP synthase deficiency reached the 'threshold' for its pathologic presentation, which we quantified at 30 %. We observed compensatory increases in the...
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The US Supreme Court, its, formation and first key decisions
Červinková, Zuzana ; Seltenreich, Radim (advisor) ; Horák, Záboj (referee)
These days, the Supreme Court of the United States represents a very important and irreplaceable role in its home land: protection of constitutionality, explanation of federal laws, as well as appellate court of last resort within the framework of American courts. Today there is no doubt about its jurisdiction, but there were doubts. In the beginning of its existence no one was sure if the Court could overcome the initial obstacles which were in the way. This diploma thesis deals with the initial existence of the Supreme Court of the United States, its role within the framework of the US federal power system, and its progressive integration as equal with the legislative branch and executive power - The United States Congress and the President, respectively. The main objective of this thesis is to introduce the federal judiciary of the United States, show its differences from the continental system, and accentuate the importance of precedents. First, I focus on the history of the United States; colonization by British citizens, the American War for Independence, The Declaration of Independence, and the formation of American statehood in the 1780s, which resulted in the acceptance of one of the longest- lasting written constitutions in the world. After the historical excurse, the thesis focuses on...
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