Polycystic Ovarian Syndrome is the one of the leading causes of infertility among women who are of child-bearing age. The syndrome’s vast range of phenotypes has made it challenging for researchers to not only consistently diagnose but also discover a cure. Currently, there are several proposed treatments being looked into, however, much of the research focuses on employing promising biomarkers, micro ribonucleic acids (miRNAs), that can potentially aid in diagnosis. The four prominent locations of research for these biomarkers include: ovarian tissues specifically looking into granulosa cells (GC), adipose tissue, follicular fluid, and the serum. My goal is to determine which of these areas holds the most promise to diagnose this syndrome in the years to come. This study reviewed a large collection of the current polycystic ovarian syndrome literature evaluating both reported miRNAs and how viable those would be as potential biomarkers to use for the future. The data showed that a majority of these promising biomarkers were found in granulosa cells, adipose tissue, and follicular fluid. Although there were miRNAs that were deemed promising in the serum, research is still far from conclusive in using these miRNAs as biomarkers for diagnosis of polycystic ovarian syndrome. By comparing the miRNAs selected from each type of location, I was able to conclude that miR-21, miR-93, miR-223, and miR-let-7b hold the most promise for the potential to become biomarkers for polycystic ovarian syndrome in the near future. Currently, there is a lot of research particularly surrounding these miRNAs and how they were shown to have been expressed in statistically significant levels among women with the syndrome. However, because of their complexity, miRNAs do not regulate one single pathway, it is hard to describe a mechanism that explains the pathophysiology of the syndrome. I believe we are still far away from successfully zooming in on one biomarker. By determining the most potential biomarker(s), we can focus resources and efforts towards finding a better diagnostic tool for this syndrome.

Background: Hyperhomocysteinemia (HHcy) is an established risk factor for thrombotic diseases yet the underlying mechanism remain unclear. In this study we investigated the effect of HHcy on endothelial cell-platelet interaction and its role in thrombosis. Methods and Results: We used a novel mouse model of HHcy (plasma homocysteine, Hcy 80 micromolar) in which a Zn2+ inducible human cystathionine beta-synthase (CBS) transgene was introduced to circumvent the neonatal lethality of the Cbs gene deficiency (Tg-hCBS Cbs-/- mice). Hcy-lowering therapy was performed by giving ZnSO4 water to induce human CBS transgene expression in adult mice. Thrombus formation was examined by photo dye-induced cremaster microvasculature thrombosis using intravital microscopy, in which endothelium was preserved, and by FeCl3-induced carotid artery thrombosis, which denudated the endothelium. HHcy accelerated cremaster arteriolar thrombosis and decreased blood flow cessation time from 41.8 min in control mice to 30.5 min in TghCBS Cbs-/- mice. Venular blood flow cessation time was slightly decreased from 5.6 to 5.0 min. Hcy-lowering therapy reduced Hcy level from 80 micromolar to 6.8 micromolar after 2 weeks of ZnSO4 water and prolonged arteriolar blood cessation time from 30.5 to 37.8 min. Interestingly, FeCl3-induced carotid artery thrombosis did not change the occlusion time. Hcy did not potentiate the aggregation and secretion function in washed human platelets from healthy donor treated with Hcy (50, 100 micromolar) or from Tg-hCBS Cbs-/- mice. However, inter-cellular adhesion molecule 1 (ICAM-1) levels, but not vascular adhesion molecule 1 (VCAM-1), were increased in cremaster tissues from Tg-hCBS Cbs-/- mice by western blot. In cultured human umbilical vein ECs (HUVEC), Hcy (100 micromolar, 24h) promoted human platelet adhesion by 200% in static adhesion assay. Using western blot, FACS and RT-PCR, we found that Hcy increased protein and mRNA levels of ICAM-1, but not that of VCAM-1, in HUVEC. ICAM-1 blocking antibody partially reversed Hcy increased platelets adhesion to HUVEC. Hcy induced ICAM-1 expression and reduced DNA methylation on ICAM-1 promoter, which were mimicked by DNA methyltransferase inhibitor azacytidine, and histone deacetylase inhibitors sodium butyrate and trichostatin A. Hcy treatment also increased intracellular Hcy, Sadenosylhomocysteine (SAH) accumulation and decreased SAM/SAH ratio in HUVECs. Hcy decreased methyl CpG binding protein 2 (MeCP2) binding and increased acetylated histone H3 (AcH3) binding to ICAM-1 core promoter region using chromatin immunoprecipitation. Pyrosequencing of ICAM-1 core promoter and adjacent region shows a decreased DNA methylation by Hcy treatment. In high methionine diet-induce HHcy in WT and Icam-/- mice, Icam-/- mice fed with HM diet only show moderately accelerated venular and barely accelerated arteriolar occlusion time compared with WT mice with CT diet using photo dye-induced thrombosis model. Conclusion: HHcy accelerates arteriolar thrombosis and increases EC-platelet interaction via ICAM-1 induction partially through DNA hypomethylation. / Pharmacology

The thesis deals with the analysis of the phase feeding in herds of Holstein cattle in specific operating conditions and its relation to milk yield and health of dairy cows. In the reference event to be optimized ration drystanding cows and lactating cows in comparison protocols laboratory analysis of mixed rations to standard NRC by the enterprise uses for compiling rations. The thesis was evaluated the effect of ration on milk yield and content of milk components and also affect the incidence of metabolic disorders. The aim of the thesis was to analyze the phase nutrition of dairy cows in the reporting company in relation to milk production and health, to evaluace the quality of feed rations, some feed and feeding technique. It was found that the monitored parameters do not match any breed mixed rations used in the standard, which could lead to reduced levels of production and reproduction and deterioration in health of dairy cows. It was further found that in the observed rearing occur very frequent occurrence of metabolic disturbances and fluctuations in the milk production.

The aim of this thesis is to identify innovation opportunities, which will serve to encourage individuals with inborn metabolic disorder phenylketunuria (also PKU) to comply with their strict dietary restrictions. The theoretical part of the thesis is devoted to the definition of basic concepts of innovation and techniques of Design thinking focusing on the Human-centered design approach. Futhermore the reader will find introduction into inborn metabolic disorder phenylketonuria. The practical part contains a plan of research, description of the current situation on the Czech market, outputs of research with experts, outputs of the workshop with individuals with PKU and the identification of innovation opportunities and recommendations.

Background: This study was conducted to determine the association between fiber intake and insulin resistance in 264 women using a cross-sectional design. Methods: Insulin resistance was indexed using HOMA-IR (fasting insulin (μU/mL)×fasting glucose (mg/dL)/405). HOMA-IR values were log transformed. Fiber and energy consumption were assessed using 7-day weighed food records. Fiber was expressed as grams per 1000 kilocalories. Body fat percentage (BF%) was measured using the BOD POD and physical activity (PA) was ascertained using Actigraph accelerometers worn for 7 consecutive days. Results: (Mean±SD) age: 40.1±3.0 years, glucose: 86.7±5.9 mg/dL; insulin: 7.1±4.3 μU/mL; HOMA-IR: 1.5±1.0; fiber intake (g/1000 kcal), total: 9.3±2.9; soluble: 1.7±0.9; insoluble: 3.8±1.9; physical activity: 2.7044 ±0.7842 million counts; BF%: 31.7±6.9; weight (kg): 66.1±10.1; total caloric intake per day (kcal): 2054.1±320.9; and dietary fat intake (% of total kcal): 30.5±0.5. Women with high total fiber intakes had significantly less insulin resistance than their counterparts (F=4.58, p=0.0332), and women with high soluble fiber intakes had significantly lower levels of insulin resistance than other women (F=7.97, p=0.0051). Participants with high insoluble fiber intakes did not differ from their counterparts (F=0.7, p=0.6875). Adjusting for either PA or BF% weakened the relationships significantly. Controlling for BF% nullified the total fiber-HOMA-IR link (F=1.96, p=0.1631), and attenuated the association between soluble fiber and HOMA-IR by 32% (F = 6.86, p=0.0094). To facilitate interpretation of the results, odds ratios were calculated to determine the association between fiber intake and HOMA-IR with both variables treated as categorical. To create dichotomous variables, fiber intake and HOMA-IR were each divided into two categories using the median (Low and High). In women who had high soluble fiber intake (upper 50%), the odds of having an elevated HOMA-IR level was 0.58 (95% CI: 0.36-0.94) times that of women with low soluble fiber intake (lower 50%). And after controlling for all of the potential confounding factors simultaneously, the odds ratio was 0.52 (95% CI: 0.29-0.93). Conclusion: High fiber intake, particularly soluble fiber, is strongly related to lower levels of insulin resistance in women. Part of this association is a function of differences in PA and BF%.

The aim of this diploma thesis was to assess the current knowledge of the pupils of 8th and 9th grades of primary schools about selected chronical non-infectious diseases (known as civilisation diseases). The pupils of rural and urban types of primary schools have taken part in the questionnaire survey. The theoretical part is focused on the characteristics of selected diseases, their causes and prevention. In the practical part, the results of the survey are processed and evaluated.

This thesis aims to investigate fluoxetine, a widely prescribed SSRI antidepressant, for its role in the pathogenesis of NAFLD and uncover novel mechanisms by which it may contribute to drug-induced steatosis. We demonstrated that increased hepatic lipid accumulation was mediated, in part, via elevated serotonin production. The inhibition of hepatic serotonin synthesis prevented lipid accumulation in fluoxetine-treated hepatocytes demonstrating a causal role for serotonin in fluoxetine-induced hepatic steatosis. Interestingly, in several studies, serotonin signaling has been shown to impact prostaglandin biosynthesis. As prostaglandins have been implicated in the development of NAFLD, and fluoxetine has previously been shown to alter the production of prostaglandins I assessed the role of prostaglandins in fluoxetine-induced hepatic lipid accumulation. Fluoxetine treatment increased mRNA expression of prostaglandin biosynthetic enzymes, increased production of prostaglandin 15-deoxy-Δ12,14PGJ2 (PPARG agonist), and elevated PPARG targets involved in fatty acid uptake. Fluoxetine-induced lipid accumulation, 15-deoxy-Δ12,14PGJ2 production, and the expression of PPARG lipogenic genes were attenuated with a PTGS1 specific inhibitor. Taken together these findings suggested that fluoxetine-induced lipid accumulation was mediated via PTGS1 and its downstream product 15-deoxy-Δ12,14PGJ2. Given that Pparg was elevated following fluoxetine treatment, and PPARG regulates microRNA involved in hepatic lipid accumulation, my final project focused on PPARG’s role in altered miRNA expression. Indeed, fluoxetine treatment increased the miRNA expression of miR-122, an effect that was attenuated when fluoxetine treatment was combined with the PPARG antagonist GW9662, suggesting a fluoxetine-PPARG-miR122 axis contributing to hepatic steatosis. While these studies have only been performed in vitro, an understanding of the molecular changes associated with SSRI treatment may lead to the development of strategies to prevent the increased risk of adverse metabolic outcomes associated with the use of SSRI antidepressants. / Dissertation / Doctor of Philosophy (Medical Science) / In adults, major depressive disorder (depression) is one of the most common psychiatric illnesses. Recent data suggests that there are more than 4.1 million Canadians who currently suffer from depression. Depression is commonly treated using selective serotonin reuptake inhibitor (SSRI) antidepressants. While these antidepressants do help manage depressive symptoms, they can also cause unwanted side effects including a build-up of fat in the liver, leading to fatty liver disease. The goal of my research is to understand the link between SSRI use and the development of fatty liver disease. This thesis investigates the effects of fluoxetine (Prozac®), a commonly used SSRI antidepressant, on molecular pathways that can lead to the development of fatty liver disease. An understanding of the molecular changes with SSRI treatment may lead to the development of strategies to prevent the harmful effects of SSRI antidepressants on the liver.