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Study of effects of prolonged administration of emtricitabine on expression of ABC efflux transporters in maternal and fetal organs
Havlová, Ivana ; Červený, Lukáš (advisor) ; Hofman, Jakub (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Ivana Havlová Supervisor: PharmDr. Lukáš Červený, Ph.D. Title of diploma thesis: Study of effects of prolonged administration of emtricitabine on expression of ABC efflux transporters in maternal and fetal organs The aim of this thesis was to evaluate the effect of chronic administration of antiretroviral substance emtricitabine (FTC) on expression of drug efflux ABC-binding cassette (ABC) transporters in the placenta, maternal and fetal organs (brain, liver, kidney, small intestine) of pregnant rats. We focused on two most important representatives of drug efflux ABC transporters, P-glycoprotein (MDR1) and Breast Cancer Resistance Protein (BCRP). Knowledge of FTC effects on gene expression of these transporters is crucial especially when using FTC in combination therapy, known as Highly Active Antiretroviral Therapy (HAART). Changes in MDR1/BCRP expression may lead to significantly altered pharmacokinetics of concomitantly administered drugs. First, bioavailability was analyzed. AUC of FTC (3,5 mg/kg) following i.m. administration was comparable to that one after i.v. administration allowing us to use i.m. application as a route of FTC administration to rats. Subsequently,...
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Gene expression of specific parts of CNS in a rat model for transgenerational transmission of drug addiction.
Radilová, Kateřina ; Černá, Marie (advisor) ; Rothová, Olga (referee)
Methamphetamine (MA) is a drug often abused by drug-addicted women and also is the mostly used drug in the Czech Republic. This drug crosses the placental and hematoencephalic barriers easily and therefore it can negatively affect not just mother herself, but also the development of the fetus. Even though there have been several published studies evincive negative effects of MA to posterity of drug-addicted mothers, the most critical term during pregnancy for the fetus remains unknown. In this study are examined CNS structures, which are directly influenced by methamphetamine action. Their function may lead to drug addiction through pleasurable feelings, such as reward feeling evoking. In the next chapter are analysed neurotransmitters and their transporters, which are related to methamphetamine effects. Studying their structure, localization or development during life may help to bring near the drug functioning in different life periods. Methamphetamine itself and its effects are closer described in the last part of this study. The problem of methamphetamine exposed fetus is outlined there - prenatal and postnatal so far known differences, but also sex differences in methamphetamine pharma- cokinetics.
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The effect of diabetes mellitus on reproductive parameters and expression of selected testicular genes in diabetic mice
Valášková, Eliška ; Pěknicová, Jana (advisor) ; Kaňka, Jiří (referee)
According to the World Health Organization (WHO), 15% of couples in reproductive age suffer from infertility problems, and up to 60% of cases are caused by male factor. Causes of this condition could be genetic background, environmental factors and various diseases, including diabetes mellitus (DM). The aim of this study was to investigate the effects of DM on reproductive parameters and expression of selected testicular genes using mouse model (FVB inbred mouse strain). DM (type 1) was artificially induced by chemical substance streptozotocin, which causes destruction of pancreatic β cells. These mice were exposed to diabetic condition for 6 weeks and then subjected to analysis. Our results have shown that diabetic condition had an impact on body weight, weight of reproductive organs as well as kidneys and livers. We also observed decreased concentration and viability of diabetic sperm compared to control. Moreover, we noticed increased staining with apoptotic marker annexin V. Further, we evaluated changes of sperm nuclear proteins - protamines. In diabetic animals, we observed higher number of sperm with insufficient protamination. Nevertheless, protamine 1 to protamine 2 ratio (P1/P2), a marker of male fertility, was not altered in sperm of diabetic animals compared to control. Regarding the...
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MicroRNAs in Human Cancers Associated with Viral Infections
Dvořáková, Lucie ; Tachezy, Ruth (advisor) ; Drda Morávková, Alena (referee)
MicroRNA (miRNA) are short single-stranded RNAs that do not encode proteins. Their main function is the regulation of the gene expression on the level of translation. This regulation is mediated by the binding of miRNA to the partially complementary segments of mRNA, both cellular or viral. It is estimated that miRNAs affect expression of at least one third of human genes and thereby influence regulation of cellular growth, differentation and apoptosis of cells. Recently the miRNAs encoded mainly by DNA viruses were discovered. These miRNAs enhance the persistence of viral infection in the host and can contribute to malignant transformation. However, the oncogenesis is also significantly affected by the regulation of cellular miRNAs expression by viral proteins. The miRNA research is topical. MiRNAs are considered as potential biomarkers and their utilization as a cancer therapy is being intensely explored. In this thesis, I'm describing the biogenesis and regulatory functions of miRNAs. I'm also presenting an overview of viral miRNAs focusing on human oncogenic viruses which do not only code their own miRNAs but also influence the expression of the host miRNAs. Finally, I am focusing on current clinical applications of miRNA. Key words: viral miRNA, cellular miRNA, oncogenesis, viral infections,...
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Gene expression of selected genes in heart of long-term cold acclimated rat under mild conditions
Marvanová, Aneta ; Žurmanová, Jitka (advisor) ; Nováková, Olga (referee)
Beneficial effect of hardening or cold adaptation for human health has been known for many decades. Beneficial or detrimental effect depends on the regimen and intensity of the adaptation and thus it is very important to pay attention to them. It has been shown in previous studies that cold adaptation of animals at a low temperature 0-4 žC is accompanied by dehydration and the damage of kidneys. As our best knowledge, the effect of mild cold adaptation on the molecular mechanisms in the heart has not been studied until now. The first aim of this study was to set up a new model of cold adaptation without any negative effects on the organism. The second aim was to assess mRNA levels of selected genes related to the important mechanisms associated with energy metabolism and cardio-protective signaling pathways in the left ventricle. Results obtained using the new model for cold adaptation at 102 řC did not show any negative changes in the blood or urine parameters and body fluid balance was maintained. Screening of mRNAs showed that chronic adaptation might affect the phenotype of the heart, transcriptional activity of the antioxidant system and of the pro-apoptotic genes, lowers adrenergic signaling and moderate glucose metabolism in cardiomyocytes. Key words: cold adaptation, heart, gene...
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Finalizing the full genome sequence of epidemic strain Burkholderia cenocepacia ST32 and identification of a prognostic marker for infections that are caused by the ST32 strain in patients with cystic fibrosis
Vavrová, Jolana ; Dřevínek, Pavel (advisor) ; Wiedermannová, Jana (referee)
Burkholderia cenocepcia is one of the serious infectious agents of respiratory tract among cystic fibrosis patients. There are problems mainly with strains which are capable of epidemic spread. The known epidemic in the Czech Republic was caused by ST32 strain in the past. In this work, there was completed whole genome sequence of referential isolate 1232 of B. cenocepacia ST32 in cooperation with bioinformatics by new generation sequencing techniques and by determining the problematic areas by a combination of Sanger sequencing bioinformatics approaches and manual assembling of sequence reads localized in these areas. The final version of the genome sequence was annotated by PGAAP and at the present time it is finalized. Second part of this work is dedicated to looking for a prognostic marker of infection caused by ST32 strain in patients with cystic fibrosis. We analysed the results of ST32 trancriptomic experiment and chose genes possibly connected with the cepacia syndrome - serious, mostly fatal state of infection. By quantitative PCR we compared their expression in isolates from 4 patients from time of cepacia syndrome and month before that. We checked the possibility of direct detection of the expression of these genes in clinical material. We identified genes for type III secretion system as...
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Relation of tumor genotype and phenotype to diagnosis, prognosis and prediction of colorectal cancer
Pitule, Pavel ; Králíčková, Milena (advisor) ; Vožeh, František (referee) ; Slabý, Ondřej (referee)
Colorectal cancer is one of the most common type of malignity. Despite of the existence of numerous studies focused on this carcinoma, there are still many unknown features regarding its diagnosis, treatment or prognosis. In the thesis we focused on the identification of novel prognostics markers that could be useful for the stratification of patients based on the disease outcomes. In the first study we immunohistochemically assessed expression of two proteins associated with cancer stem cells in the samples of primary colorectal cancer and matched liver metastasis. Goal of the study was to evaluate relation among expression of CD44 and CD133 and overall survival and disease free interval in our set of patients. We observed that increased ratio of CD133 positive compared to CD133 negative tumor glands resulted in longer disease free interval, finding which is opposite to the general view on the CD133 role in the cancer development. Our hypothesis is that we analyzed confined group of patients and followed a bit different goal, where we measured ratio between positive and negative glands in the view-field and not the intensity of staining as the previous studies did. Our second study was focused on the transcriptional analysis of the selected set of twelve genes using frozen samples from colorectal...
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Regulation of bacterial transcription by alternative sigma factors.
Benda, Martin ; Krásný, Libor (advisor) ; Roučová, Kristina (referee)
An important feature of bacteria is their ability to respond to various environmental conditions by regulation of transcription. This thesis is focused on regulation of transcription initiation by different sigma factors. Sigma factor is a specific subunit of RNA polymerase, which ensures correct recognition of promotor sequences. During exponential growth phase under ideal conditions, the cell transcribes most of the genes under the control of the so-called housekeeping sigma factor. In the transition to stationary phase, during sporulation or upon exposure to different stresses, many genes are activated and transcribed under the control of alternative sigma factors. This work presents a list of these alternative sigma factors of Bacillus subtilis, focusing on conditions under which the individual sigma factor is used, methods of activation and repression of these factors and on regulons controled by these sigma factors.
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The Role of oncogenic microRNA - 155 and proto - oncogen MYB in chronic lymphocytic leukemia
Vargová, Karina
(EN) Chronic lymphocytic leukemia (B-CLL) represents a disease of mature-like B-cells. Due to failed apoptosis but also due to enhanced proliferative signals, the leukemic B-cells accumulate in the peripheral blood, bone marrow, lymph nodes and spleen. The clinical course of B-CLL is very heterogeneous; in some patients B-CLL progresses very rapidly into an aggressive form. Such patients need therapy sooner while in other patients with indolent B-CLL the onset of therapy takes years. Several standard prognostic and disease progression markers are used for disease staging and monitoring, however a reliable marker that will suggest when to start therapy is unknown. Expression of small, non-coding microRNAs is often deregulated and represent important prognostic markers in variety of cancers including leukemia. Hence in our study we concentrated to miR-155, an important molecule regulating differentiation of hematopoietic cells, inflammation process and antibody production. Its aberrant expression was described in Hodgkin`s as well as in non-Hodgkin`s lymphoma, including indolent lymphoproliferations like B-CLL. Our results confirmed elevated levels of both, primary miR-155 transcript and mature form of miR-155 in our B-CLL patient samples (N=239). The aberrant expression of miR-155 in B-CLL samples...
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Analysis of gene expresion via next generation sequencing techniques
Bláhová, Monika ; Krylov, Vladimír (advisor) ; Žárský, Vojtěch (referee)
1 Abstract The main task of this work is create review today's available methods for gene expression analysis, introduce advantages and disadvantages of this metods and compare them. Nowadays sequencing is one of the most usable molecular methods. Sequencing methods are divided to three groups, a first generation sequencing, a second generation sequencing and a third generation sequencing. The most useful are the second generation sequencing. However, the third generation sequencing have a big potencial too. It is not necessary amplificate samples using PCR thanks them. Amount of data raises rapidly, thanks decreasing costs and increasing efficiency. Demands on data starage, computers output are growing. And softwares for data analysis are much clever than ever before.
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