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Expression of candidate genes for prostate cancer
Krupicová, Daniela ; Mareš, Jaroslav (advisor) ; Holá, Dana (referee)
4 Abstract Prostate cancer is one of the major medical problems within the male population in the Czech Republic and in the world. It is on second place among cancer illnesses with respect to mortality in czech male population. Its incidence strongly increases with age. Prostate cells have a unique ability to accumulate zinc in high concentrations compared to other tissues of human body. It is necessary for the proper physiological function of the prostate. There was detected loss of this accumulation ability in prostate cancer cells, which seems to be a condition to carcinogenesis in prostate cells. In this thesis was investigated the expression of four genes involved in the maintenance of homeostasis of zinc in prostate cells. Genes ZIP1 and ZIP7 encode zinc transporters, genes MT1-F and MT2 encode metallothioneins. There was collected 90 biopsy specimens from patients with prostate cancer or with benign prostatic hyperplasia. mRNA was isolated from these samples, cDNA was obtained by RT-PCR. This cDNA was detected by gel electrophoresis and the results were statistically evaluated. Several correlations was found between gene expression and the clinical data of patients. The most important result, there was found lower levels of expression of genes MT1- F and ZIP1 in samples of patients with cancer...
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Candidate genes screening for prostate cancer
Semaneková, Viera ; Mareš, Jaroslav (advisor) ; Šťovíček, Vratislav (referee)
Prostate carcinoma is considered to be one of the main medical problem in male population. Prostate carcinoma is the most frequently diagnosed malignancy in men and the death rate has the second position within all diagnosed malignancies in Czech Republic (ÚZIS). There is only one reliable diagnostic tool: PSA (prostate specific antigen). Level of PSA is often elevated in men with prostate carcinoma. This diploma thesis is focused on study of changes in gene expression in prostate carcinoma. Three candidate genes were analyzed: VCL (vinculin), SHB (Src homology 2 binding protein) and OCT3 (organic cation transporter 3). According to recent publications, these genes are related to tumor progression and they could have prognostic significance. In this thesis the following methodological approaches were used: 82 prostatic specimens were collected from patients and mRNA was isolated from these specimens; then RT-PCR was used to obtain cDNA, fragments were detected by electrophoresis. At the end statistical methods were used for evaluation. Relative expression of the genes in prostate carcinoma tissue was compared to relative expression of the genes in BPH (benign prostatic neoplasia) tissue. Results showed higher expression of genes SHB and OCT3 in prostate carcinoma tissue in compariscon to BPH...
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Biotransformation of phenols by enzymatic systems of Candida tropicalis yeast and Comamonas testosteroni bacteria
Vilímková, Lenka ; Stiborová, Marie (advisor) ; Entlicher, Gustav (referee) ; Mareš, Jaroslav (referee)
Candida tropicalis yeast and bacteria Comamonas testosteroni have been considered to be able to metabolize phenol and utilize it as the only source of carbon and energy. In our laboratory we investigated the cytoplasmic enzymes responsible for the first and second step of phenol degradation, NADPH-dependent phenol hydroxylase of both C. tropicalis and C. testosteroni and catechol 1,2-dioxygenase of C. tropicalis. The aim of our study was to isolate and partially characterize those enzymes. Phenol hydroxylase purification consisted of preparation of cytosol from C. tropicalis yeast by fraction centrifugation, chromatography and re-chromatography on a column of DEAE Sepharose, fractionation by precipitation of the enzyme with polyethylene glycol 6000 and gel permeation chromatography on a column of Sephacryl S-300. Extracellular phenol hydroxylase of C. testosteroni was purified by fraction precipitation with polyethylene glycol 6000 and by gel permeation chromatography on 4B Sepharose and Sephacryl S-300. Catechol 1,2-dioxygenase was purified using the procedure consisting of: chromatography and re- chromatography on a column of DEAE Sepharose, lyophilization of the enzyme and gel permeation chromatography on a column of Sephadex G-100. The enzyme activity was determined by two methods: use of HPLC...
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Cortical and Subcortical Mechanisms of Time Perception
Dušek, Petr ; Jech, Robert (advisor) ; Mareš, Jaroslav (referee) ; Bareš, Martin (referee)
Deficits in interval timing have been described in focal brain lesions and in various neuropsychiatric disorders including Parkinson's disease (PD). The aim of this study was to explore brain areas responsible for human time perception and for the timing deficit in PD. We used a time reproduction task (TRT) which consisted of an encoding phase (during which visual stimuli of durations from 5 to 16.6 sec were presented) and a reproduction phase (during which interval durations were reproduced by a button pressing). In our first fMRI study, we used a parametric modulated analysis searching for brain areas with activity, expressed as Blood Oxygenation Level Dependent (BOLD) signal, correlated with the duration of time interval. During the encoding phase, there was a gradual deactivation of the left prefrontal cortex (PFC) and cingulate gyrus. During the reproduction phase, there was a gradual deactivation in precuneus and an accumulation of activity in the left PFC, primary motor area, right caudate and supplementary motor area (SMA). The second study aimed at supporting the role of two of these areas, SMA and precuneus in interval timing by repetitive transcranial magnetic stimulation (rTMS). Accuracy and variability of time estimates were compared before and after rTMS. Accuracy of estimates was not...
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Study on metabolism of 3-aminobenzanthrone and induction of biotransformation enzymes
Mizerovská, Jana ; Stiborová, Marie (advisor) ; Mareš, Jaroslav (referee) ; Hýsková, Veronika (referee)
CHARLES UNIVERSITY IN PRAGUE FACULTY OF SCIENCE DEPARTMENT OF BIOCHEMISTRY Study on metabolism of 3-aminobenzanthrone and induction of biotransformation enzymes Summary of PhD Thesis RNDr. Jana Mizerovská Supervisor: Prof. RNDr. Marie Stiborová, DrSc. Prague 2010 RNDr. Jana Mizerovská Introduction 1 INTRODUCTION 3-Nitrobenzanthron, precursor of 3-ABA The nitroaromatic 3-nitrobenzanthrone (3-nitro-7H-benz de anthracen-7-one, 3-NBA) occurs in diesel exhaust and in airborne particulate matter(8, 9, 14) . 3-NBA is most likely formed during the atmospheric reaction of benzanthrone with nitrogen oxides, especially in the presence of ozone, or during imperfect burning of diesel. 3-NBA exhibits extremely high mutagenic activity(9, 14) and is also a genotoxic carcinogen causing lung tumors in rats(14) . 3-NBA is also evaluated to be a potential carcinogen for humans(14, 1, 9, 19) . The genotoxicity of 3-NBA was documented by the detection of specific 3-NBA-derived DNA adducts in vitro, in human cell lines and also in vivo in rats and mice(12, 13, 15, 2) . The predominant DNA adducts formed by 3-NBA after its metabolic activation by reduction of the nitro group are 2-(2'-deoxyguanosin-N2 -yl)-3- aminobenzanthrone and N-(2'-deoxyguanosin-8-yl)-3-aminobenzanthrone(9, 14) and these are most probably responsible for the...
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není uveden
Soukup, Viktor ; Babjuk, Marek (advisor) ; Mareš, Jaroslav (referee) ; Novák, Jaroslav (referee)
Immunohistochemical examination of microvascular density can be an additional prognostic factor in patients with superficial urinary bladder carcinoma, nevertheless, more extensive studies need to be carried out. Maximal microvascular density correlated with a recurrence free interval and in a case of bigger group of patients could be statistically significant. The statistical correlation between MVD and a nuclear p53 accumulation is that significant that we assume the p53 importance in the angiogenesis regulation of superficial urinary bladder carcinoma. The p53 positivity in non-tumor mucosa of urinary bladder in patients with superficial bladder cancer correlates with higher risk of tumor recurrence. According to our results the reason of urinary bladder carcinoma multifocality is genetic instability present in the whole urothelium at the time of tumor diagnosis. The PAX5 gene expression is a frequent finding in superficial transitional cell carcinoma of the bladder. From the prognostic point of view, high levels of PAX5 expression seem to be connected with higher recurrence and progression rates. The extent of lamina propria invasion seems to be a clinically relevant prognostic factor for progression of T1 bladder transitional cell carcinoma. Finding T1b tumor is connected with the risk of understaging....
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Microsatellite instability in hereditary nonpolyposis colorectal cancer patients
Sekowská, Martina ; Křepelová, Anna (advisor) ; Mareš, Jaroslav (referee) ; Kleibl, Zdeněk (referee)
Detection of microsatellite instability (MSI) is the standard part of mutational analysis in hereditary nonpolyposis colorectal cancers (HNPCC). Characteristic phenotypic feature of MSI indicates loss of mismatch repair (MMR) in tumor cells. We studied MSI in 205 tumors from 152 patients with HNPCC. Of these, 37 patients fulfilled Amsterdam criteria, 72 patients were familial and 43 were sporadic cases. We used methods of fragmentation analysis on polyacrylamide gel and/or with fluorescent labelled primers (ABI Prism 310 Genetic Analyzer). Three mononucleotide (BAT-RII, BAT-25, BAT-26) and five dinucleotide (D2S123, D3S1029, D5S346, D17S250, D18S58) repeat loci were analysed. We detected 75 tumors with high degree of MSI (MSI-H), 12 tumors with low degree of MSI (MSI-L) and 118 tumors with stable microsatellites (MSS). In 44 of these, loss of heterozygozity (LOH) was found. In 30 patients with MSI-H tumors a mutation in one of mismatch repair genes was detected. Microsatellite analysis was positively correlated with immunohistochemical detection of MLH1 and MSH2 proteins.
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