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Gene Expression Data Analysis for in Vitro Toxicology
Šimeček, Petr
Microarray analyses have been performed on CD34-progenitor derived dendritic cells that have been exposed to chemical skin sensitizers; allergens (4) and irritants (2). The aim of this work is to identify genes whose expression is affected by exposure to the allergens but not by the irritants. The inference is based on a dataset containing 72 microarrays.
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název v anglickém jazyce není uveden
Kolesová, Hana ; Grim, Miloš (advisor) ; Slípka, Jaroslav (referee) ; Nečas, Emanuel (referee)
Branchial arches region and its blood vessels are extensively transformed in the embryonic development. Aim of this study is to investigate mechanisms of the branchial arches region development and to study how a morphogen Sonic hed gehog (Shh) participate on the formation and remodeling of branchial arches and their blood vessels. Influence of Shh was evaluated based on the changes caused by its inhibition in vivo. Shh function was inhibited with an anti-Shh antibody, which was produced into the embryo from the applied hybridoma cells. Shh signaling cascade was also inhibited by cyclopamine. Results show that Shh is important for dc novo format,ion of the blood vessels in the branchial arches region. Further Shh is necessary for stabilization of the vessel wall, mainly for anterior cardinal vein. Shh also alfects later vessel development and transformation, which includes i.e. fusion of the dorsal aorta, branching of the internal carotid artery and outflow tract development. Short time inhibition of Shh has minor effect on the apoptosis and proliferation activity of the branchial arches region mesenchymal cells. We assume that Shh signals directly to the blood vessels endothelial cells, as Shh receptor ptel is also expressed in endothelial cells and its signal is reduced with Shh inhibition. In studied...
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Signaling pathways and genes regulating gonadotropin-induced maturation of porcine oocytes and cumulus expansion
Blaha, Milan ; Němcová, Lucie (advisor) ; Král, Jiří (referee)
In vitro, meotic maturation of porcine oocytes and cumulus expansion are induced by FSH and EGF-like peptides AREG and EREG. FSH and EGF-like peptides induce expression of cumulus expansion-related genes (HAS2, PTGS2 and TNFAIP6). To define signaling pathways that control FSH- and AREG-induced cumulus expansion, porcine cumulus-oocyte complexes were treated with specific protein kinase inhibitors. Inhibitors of MAPK3/1, MAPK14 and ERBB1 significantly reduced both FSH- and AREG-induced expression of HAS2, PTGS2 and TNFAIP6. These inhibitors decreased FSH/LH-induced expression of AREG and EREG in mural granulosa cells. Surprisingly, inhibitor of PKA had no effect on AREG expression in cumulus-oocyte complexes but the inhibitor decreased expression of TNFAIP6 induced by AREG. Inhibitor of PI3K increased expression levels of AREG and PTGS2 but EREG, HAS2 and TNFAIP6 were reduced. Expression levels of the cumulus expansion-related genes were not affected by an analog of cGMP (8-CPT-cGMP). However, 8-CPT-cGMP blocked spontaneous in vitro meiotic maturation of porcine oocytes and its effect was abolished by FSH. Key words: cumulus expansion, cumulus expansion-related genes, meotic maturation, FSH, amphiregulin, cGMP
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MDOCTM effect on expression of cell adhesion molecules / / in the vessel wall of ApoE / LDLR-deficient mice
Drnek, Ludvik ; Nachtigal, Petr (advisor) ; Štaud, František (referee)
Atherosclerosis belongs to the cardiovascular diseases, which have been the most frequent cause of the morbidity and mortality in all the economically advanced countries of Europe and the USA in the last few years. The purpose of this thesis was to find some potential hypolipidemic and antiinflammatory effects of MDOC. The new potential substance belongs to the group of dietary fibre. ApoE/LDLr-deficient mice (n=8) was used in the study. The mice were fed with aterogenic diet containing 1.25% cholesterol for 8 weeks. In MDOC group, the mice were fed on the same aterogenic diet, with 5% MDOC added to this diet. Blood sample for the biochemical investigation were taken from animals. Then, partitions of the tissue formed by the aorta together with the upper part of the heart were taken, too. Then, a biochemical analysis of the blood was made and also in aortic sinus and arch an imunohistochemical and stereological analysis of the endothelial expression of VCAM-1 and ICAM-1 were performed. The results of the biochemical analysis showed that MDOC treatment resulted in a significant decrease of total cholesterol, VLDL and significant increase of HDL cholesterol. Stereological analysis of immunohistochemical staining showed significant decrease of VCAM-1 staining in the atherosclerotic lesion after the...
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Effects of Spirulina platensis on the expression of endothelial NO synthase in apoE-deficient mouse model of atherosclerosis
Pešlová, Monika ; Nachtigal, Petr (advisor) ; Kopecký, Martin (referee)
1. ABSTRACT Spirulina platensis is a blue-green microalga belonging to the cyanobacteria family. Microalga is a large source of proteins and good fatty acid and other nutritional elements, for example iron, calcium, chromium, lithium, selenium. It also contains natural dyes chlorophyll, phycocyanin. Its antioxidant and anti-inflammatory were described. The aim of this thesis was to determine potencial hypolipidemic effects and potential effects on endothelium of Spirulina platensis in apoE-deficient mice. ApoE-deficient mice were fed standard diet for 2 weeks. At the age of 8 weeks the control group of animals were fed with the western type diet, which contained 21% fat and 0,15% cholesterol for 8 weeks. The same atherogenic diet was used in Spirulina platensis group, where Spirulina platensis was added to the atherogenic diet at the dosage of 20 mg per day. The biochemical analysis of lipid spectrum was done, area of atherosclerotic lesions was determined and imunohistochemical and stereological analysis of eNOS expression was performed as well. The results of this thesis showed positive effects of Spirulina platensis on cholesterol levels, VLDL and LDL cholesterol. Moreover stereological analysis of imunohischemical staining revealed that, that endotelial expression eNOS was significantly increased by...
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Calculus Bovis effect on blood lipids and the expression of endothelial NO synthesis / / in ApoE-deficient mouse model of atherosclerosis
Studená, Romana ; Nachtigal, Petr (advisor) ; Trejtnar, František (referee)
Calculus Bovis is one of the most precious and commonly-used medicinal materials in China. It is consists predominantly bilirubin and bile acids. In this thesis we focused on possible effects of Calculus Bovis on lipids and atherogenesis in apoE-deficient mice. ApoE-deficient mice were fed standard diet for 2 weeks. At the age of 8 weeks the control group of animals were fed with the western type diet, which contained 21% fat and 0,15% cholesterol for 8 weeks. The same atherogenic diet was used in Calculus Bovis group, where Calculus Bovis was added to the atherogenic diet at the dosage of 50 mg/kg b.wt. Calculus Bovis (Artificial bezoar, Biopharma, Australia) per day. The biochemical analysis of lipid spectrum was done, area of atherosclerotic lesions was determined and imunohistochemical and stereological analysis of eNOS expression was performed as well. Calculus bovis treatment did not affect blood lipids, total cholesterol, VLDL, LDL, HDL and TAG when compared with control mice. Moreover Calculus bovis treatment did not decrease either atherosclerotic plaque area or endothelial expression of eNOS. The failure of Calculus Bovis treatment in this pilot study could be probably related to insufficient dose of Calculus Bovis.
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Circadian rhythm of secondary alga Chromera velia
JONÁKOVÁ, Martina
A life cycle of many known algae is influenced by the circadian clock. It seems probable that also Chromera velia, the closest known photosynthetic relative to Apicomplexa, uses circadian oscillator. We tested, if fluctuating of zoospore?s abundance is driven by the circadian clock. This rhythm is stable even in the constant light condition. Furthermore, three cryptochrome genes, involved in circadian rhythms, were sequenced and consequently phylogenetically investigated. Two of them were classified as CRY-DASH, the third is CRY-DASH-like. The rhythmicity of CRY-DASH expression in C. velia was also investigated. The level of expression was also tested under constant light and dark condition. It was suggested that expression of these genes is associated with the circadian oscillator. This is the first study of the circadian clock in C. velia.
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