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Microarray Data Interpretation
Ludwig, Petr ; Šilhavá, Jana (referee) ; Smrž, Pavel (advisor)
This Bachelor thesis explains the basics of biochip or microarray data interpretation, starting with short introduction to genetics, especially genetic information significance evaluating. The focus was set mainly on the set of scripts transforming and analyzing the sample data. The data used in this thesis are a result of biochip analysis of the Colon Tumor tissues. The secondary result represents disclosing the main marker for this particular type of cancer, the primary result is evaluation of marker significance in the context of signaling pathways. The resulting pathways are sorted by relevance.
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Účinky vybraných peptidů na jednobuněčnou řasu Chlamydomonas reinhardtii
Avdeeva, Olga
In the presented work, the modified novicidin peptides NVCAA and NVCHH were applied to the unicellular alga Chlamydomonas reinhardtii. The aim of the study was to determine the minimum inhibitory concentrations of the tested peptides on Escherichia coli bacteria with subsequent application on the unicellular algae Chlamydomonas reinhardtii. The assessment of the effect of the tested peptides on the algae was carried out using spectrophotometric analyses: chlorophyll A and chlorophyll B content, total flavonoid content, total antioxidant capacity and using the DPPH ASSAY method. Chlamydomonas reinhardtii molecular changes were determined by monitoring the amount of isolated RNA and the expression of APX1, APX2, CAT1, CAT2 genes using the Real-time PCR method. Statistically processed data showed that the application of NVCAA and NVCHH peptides induced considerable stress in the cilia.
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The effect of cancerogenic azo dye Sudan I on expression of biotransformation enzymes
Hejduková, Žaneta ; Svášková, Dagmar (advisor) ; Dračínská, Helena (referee)
Sudan I is a widely used azo dye which has the ability to cause carcinomas in organs and tissues of experimental animals. During reactions catalyzed by microsomal monooxygenase enzyme systems or cytoplasmic biotransformation enzymes, Sudan I is oxidized to reactive metabolites that covalently bind to nucleic acids and cause their damage. Sudan I can also be metabolized by reduction, e. g. by a DT-diaphorase enzyme (NQO1). Reduction of Sudan I is considered to be a detoxification reaction. In this work, the in vivo action of Sudan I is examined in terms of its ability to induce an expression of the biotransformation enzyme DT-diaphorase in tissues of rats treated with the azo dye. The aim of this work was to quantify the degree of NQO1 induction at mRNA level. After the isolation of total RNA from organs of rats treated with Sudan I, the RNA was converted to cDNA by reverse transcription using random hexamers as primers. Using specific probes, the abundance of mRNA for the enzyme NQO1 in the organs of treated rats was quantified by "real-time" PCR, relatively to the control gene with a constant expression (β-actin). Through comparing thus determined amounts of mRNA in individual organs of treated and untreated rats, it has been found that Sudan I had caused a significant increase in the expression...
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Laccase activity profiling in Trametes versicolor cultures degrading endocrine-disrupting compound Delor 103
Plačková, Martina
In this work endocrine disrupting potential of Delor 103, a commercial mixture of PCB congeners, was studied along with its effect on production of laccase by the ligninolytic fungus Trametes versicolor. Using a gene-reporter yeast assay for evaluation of hormonal activity Delor 103 showed an androgenic activity with an EC50 value of 2.29. 10-2 mg/l. Chlorbenzoic acids, Delor 103 potential metabolites resulting from microbial degradation, displayed on the other hand an estrogenic activity, indicating possible changes in hormonal activity of Delor 103 during its microbial degradation. The addition of Delor 103 to mineral medium T. versicolor cultures resulted in an up to 257times higher laccase activities detected in fungal cultures. Delor 103 induced enzymes showed different pI values from those of control cultures. In a complex malt-extract glucose medium (MEG) the stimulation effect of Delor 103 was kept down. Further, the production of laccase and synthesis of different pI forms depended strongly on the growth phase of fungal cultures. Exponencially growing cultures of T. versicolor were able to produce up to 7 different pI forms of laccase in responce to Delor 103 whereas stationary cultures produced only 4 enzyme forms with higher pI values. Stimulation of laccase activities in T. versicolor,...
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Vliv olejů s rozdílným složením polynenasycených mastných kyselin na vybrané imunologické markery u modelového organizmu
Křikavová, Radka
The aim of this thesis was to observe how polyunsaturated fatty acids affect the wound healing process. This effect was observed by expression of selected genes and collagen type I and III in the model organism. The Wistar Albino laboratory rat strain was chosen as a model organism. 50 rats were used for the experiment. Rats were divi-ded into five groups of ten. One control group was selected which was fed only with compound feed without the addition of oil. The rats in the remaining groups were fed a feed mixture with the addition of oil (Schizochytrium extract oil, fish oil, palm oil and safflower oil). After 52 days of fattening, rats under anesthesia were excised on the dor-sal side of the back. Fattening was continued for 12 days and then rats were sacrificed with isoflurane to remove liver and healing skin wounds. EPA / DHA deposition was determined from collected liver. RNA was obtained from healing skin wounds for quan-titative PCR with specific primers for TGF-β1, PTGS2, ACTA2, VEGF, COL1A1, COL3A1. Further, immunohistochemical sections with type I and III collagen monoclo-nal antibodies were generated from the healing skin wound. The assumption of achie-ving better results with high DHA oil was not lost. Best results in the wound healing process were achieved in rats fed with safflower and fish oil (p <0.05). These results are inconsistent with the literature, since it was assumed that only n-3 PUFAs contribute to a better wound healing process and n-6 PUFAs, on the other hand, prolong wound hea-ling. Ambiguous results regarding the use of n-3 PUFAs in the wound healing process require further research.
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