|
|
Study of extracellular placental specific microRNAs in maternal circulation and their utilization in clinical diagnostics of pregnancy-related complications
Kotlabová, Kateřina ; Hromadníková, Ilona (advisor) ; Černá, Marie (referee) ; Korabečná, Marie (referee)
Objectiees: Our etudy inieetigated the poeeible utliiaton of extracellular microRNAAe ae noiel non-iniaeiie biomarkere for diaignoetce of preignancy-related complicatone euch ae igeetatonal hyperteneion (GH), preeclampeia (PE) and fetal igrowth reetricton (FGR). Firet, we identifed appropriate preignancy-aeeociated (placenta epeciifc) microRNAAe in maternal circulaton in preignanciee with normal couree of igeetaton. Then, we quantifed eelected extracellular C19MC microRNAAe in maternal circulaton oiertme in normally proigreeeinig preignanciee. Subeequently, we compared C19MC microRNAA expreeeion proiflee in maternal circulaton between preignanciee with clinically eetabliehed preignancy-related complicatone (PE, FGR, GH) and igeetatonal-aige-matched controle. Finally, monitorinig of eelected placenta epeciifc C19MC microRNAAe in maternal circulaton within the ifret trimeeter of igeetaton wae performed with the aim to identfy extracellular C19MC microRNAAe able to diferentate between normal preignanciee and thoee at riek of eubeequent deielopment of preignancy- related complicatone. Reeultes: The leiele and expreeeion proiflee of extracellular placental epeciifc microRNAAe in circulaton of non-preignant indiiiduale and preignant women were performed ueinig real- tme RT-PCR. The eelecton of...
|
|
|
Prognosis and diagnostics in acute myeloid leukemia
Jiráková Trnková, Zuzana ; Jíra, Milan (advisor) ; Černá, Marie (referee) ; Špíšek, Radek (referee)
The main aim of the dissertation was in patients with acute myeloid leukemia to investigate the impact of favorable prognostic chromosomal aberrations and increased expression of a family of multiple drug resistance genes to achieve a complete remission and overall survival, and identify the potential prognostic factors. Experiments were focused on the introduction of routine diagnostics of two prognostically favorable fusion genes, AML1/ETO and CBFbeta/MYH11 and multiple drug resistance gene MDR1. This testing was performed by reverse transcription, PCR and subsequent electrophoretic analysis of its products.
|
|
|
Epigenetic regulation of HLA class II genes and their role in autoimmune diseases
Čepek, Pavel ; Černá, Marie (advisor) ; Štechová, Kateřina (referee) ; Reiniš, Milan (referee)
(EN) Type 1 diabetes (T1D) belongs among polygenic multifactorial autoimmune diseases. The highest risk is associated with HLA (human leukocyte antigen) class II genes, including HLA-DQA1 gene. Our aim was to investigate DNA methylation of HLA-DQA1 promoter alleles (QAP) and correlate methylation status with individual HLA-DQA1 allele expression of T1D patients and healthy controls. DNA methylation is one of the epigenetic modifications, that regulate gene expression and is known to be shaped by the environment. 61 T1D patients and 39 healthy controls were involved in this study. Isolated DNA was treated with sodium bisulfite and HLA-DQA1 promoter sequence was amplified using nested PCR. After sequencing, DNA methylation of HLA-DQA1 promoter alleles was analyzed. Individual mRNA HLA-DQA1 relative allele expression was assessed using two different endogenous controls (PPIA, DRA). We have found statistically significant differences in HLA-DQA1 allele 02:01 expression (PPIA normalization, Pcorr=0.041; DRA normalization, Pcorr=0.052) between healthy controls and T1D patients. The complete methylation profile of the HLA-DQA1 promoter was gained with the most methylated allele DQA1*02:01 and the least methylated DQA1*05:01 in both studied groups. Methylation profile observed in T1D patients and healthy...
|
|
|
Genetic backround and new biochemical markers in pathological pregnancy
Germanová, Anna ; Kalousová, Marta (advisor) ; Černá, Marie (referee) ; Bílek, Radovan (referee)
Receptor for advanced glycation end products (RAGE), its soluble form (sRAGE) and glyoxalase 1 (GLO 1) are important part of pathogenesis of many chronic diseases. The aim of this thesis was to elucidate role of sRAGE, four chosen RAGE polymorphisms and one GLO 1 polymorphism in physiologic pregnancy and in pregnancy with complications. Serum sRAGE levels were determined in healthy pregnant women (N=120) and in pregnancies complicated with threatening preterm labor (N=99), preeclampsia (N=35), intrauterine growth restriction (IUGR) (N=22) and intrahepatic cholestasis of pregnancy (ICP) (N=14). Four RAGE polymorphisms (RAGE -429T/C, -374A/T, RAGE Gly82Ser (557G/A), RAGE 2184A/G) and one glyoxalase 1 polymorphism GLO 1 Glu111Ala (419A/C) were studied in the same population of healthy pregnant women and women with pathological pregnancy. Serum sRAGE levels are low in comparison to non-pregnant controls, but they vary during the physiologic pregnancy. Serum sRAGE levels are low in the 1st trimester, increased in the 2nd trimester and again decreased in the 3rd trimester. Women with premature labor have significantly decreased serum sRAGE levels in comparison to women with threatening premature labor and in comparison to healthy pregnant women. sRAGE correlates negatively with leukocyte count in preterm...
|
|
|
Pregnancy proteins - molecular biological and biochemical analysis
Muravská, Alexandra ; Kalousová, Marta (advisor) ; Černá, Marie (referee) ; Průša, Richard (referee)
The aim of this thesis was to establish methods for selected PAPP-A (Pregnancy- Associated Plasma Protein A) gene polymorphisms analysis and to study genetic background of PAPP-A and biochemical background of PAPP-A and PlGF (Placental Growth Factor) in relation to risk pregnancy. Secondly, the aim was to establish method for two-dimensional (2D) electrophoresis of amniotic fluid. Methods for analysis of ten PAPP-A gene polymorphisms were established. These polymorphisms, PAPP-A and PlGF levels were studied in together 165 women in third trimester pregnancies complicated with threatening preterm labor (n=98), preeclampsia (n=35), IUGR (Intrauterine Growth Restriction) (n=34) and ICP (Intrahepatic Cholestasis of Pregnancy) (n=15). 114 healthy pregnant women served as controls. The method for 2D electrophoresis of amniotic fluid was established. Preeclamptic patients had significantly higher frequency of TT genotype of Cys327Cys (C/T) PAPP-A gene polymorphism compared to controls. Patients with ICP had increased serum levels of PAPP-A compared to controls, in patients with threatening preterm labor PAPP-A levels were rather decreased. PlGF levels did not differ from control group in patients with ICP and threatening preterm labor. Positive correlation was found between PAPP-A and PlGF in group of...
|
|
|
Nutrigenomic analysis of diet influence in prenatal and early development on metabolic syndrome aspects manifestation in adulthood.
Školníková, Elena ; Šeda, Ondřej (advisor) ; Černá, Marie (referee) ; Malínská, Hana (referee)
16 Abstract The rising prevalence in noncommunicable diseases worldwide calls for the effort to determine their underlying causes. Common metabolic disorders in particular overwhelm the healthcare systems and are a one of the leading causes of poor quality of life of patients. Metabolic syndrome is represented by concurrence of several conditions - dyslipidaemia, obesity, hypertension or impaired glucose tolerance - altered metabolic phenotypes related to genetic and environmental factors. Recent studies suggest that early-life exposure to certain environmental stimuli is particularly capable of changing the mammalian phenotypes. Nutrition, as one of the major factors influencing health, is naturally a focus of research, which studies the link between parental diets and phenotypic alterations in offspring. The developmental origins of health and disease were historically more focused on maternal undernutrition, it is, however, more important to focus on surplus of macronutrients considering the westernization of modern diets. We propose the relevancy of not only the amount of macronutrients in maternal diet, but also their sources, as they may increase disease risk in offspring. Here we report, that sucrose as an alternative carbohydrate in maternal diet, has a marked impact on metabolism of the offspring...
|
|
|
Molecular biomarkers related to DNA damage and repair: their role in carcinogenesis, patients' treatment and monitoring
Vodenková, Soňa ; Vodička, Pavel (advisor) ; Anděra, Ladislav (referee) ; Černá, Marie (referee)
Genome instability represents one of the leading forces driving the onset and development of cancer. It arises as a consequence of the combined effect of DNA damage and errors made by the DNA repair system. In many cancers, DNA damage tolerance and DNA repair pathways are disrupted or deregulated, thereby promoting cancer progression. DNA repair also appears to play a substantial role in cancer therapy response. This Dissertation Thesis was performed in response to several unclear and unresolved issues of the role of DNA damage and DNA repair in cancer pathogenesis. The aim of the Thesis was to search for potential novel biomarkers and confirmation of the validity of already existing biomarkers related to DNA damage and DNA repair, which may be associated with cancer susceptibility and patient's clinical outcome. We also explored the biological basis of different biomarkers and their associations. The major outcomes of this Thesis are: 1) The elevated chromosomal aberrations (CAs) in peripheral blood lymphocytes (PBLs) may serve as a biomarker of cancer susceptibility and partially affects patients' clinical outcome. While telomere shortening contributes to the formation of CAs in PBLs only in healthy individuals, less efficient DNA double- strand break repair in PBLs is associated with telomere...
|
|
|
Immunoregulatory characteristics of immune cells of children of allergic and non-allergic mothers and the possibility of their modulation with probiotic E. coli strain O83:K24:H31
Černý, Viktor ; Hrdý, Jiří (advisor) ; Tlaskalová - Hogenová, Helena (referee) ; Černá, Marie (referee)
Due to high incidence, medical and socioeconomic burden and impact on individual quality of life and productivity, allergic disorders are a crucial issue for 21st century immunology. Much still remains to be elucidated, particularly regarding the very early processes in allergy development. In order to introduce timely, effective preventive measures, novel, more reliable predictive factors of allergy risk also need to be established. Dysregulation of proper balance between the branches of immune response, particularly unwarranted dominance of Th2, is the underlying cause of allergy. After birth, new immune balance needs to be established to prepare the neonate for adequate reactivity towards newly encountered environmental stimuli. Regulatory T cells (Treg) play a central role in finely setting this balance and inducing tolerance towards harmless environmental antigens, including allergens. Interactions with external factors, most importantly microbiota, modulate this process during the early postnatal "window of opportunity." Analysis of cord blood Treg of children of allergic mothers uncovered decreased presence of function-associated surface markers and lower production of IL-10. Furthermore, decreased proportion of Helios- induced Treg was observed in children with higher risk of allergy....
|
|
|
TET1 overexpression, DNA hypomethylation and aberrant expression of human endogenous retrovirus ERVWE1 in germ cell tumors
Benešová, Martina ; Trejbalová, Kateřina (advisor) ; Černá, Marie (referee) ; Reiniš, Milan (referee)
TGCTs are tumors of male germ cells. They comprise of seminomas and non-seminomas (embryonal carcinoma, yolk sac tumor, choriocarcinoma, and teratoma). GCT types differ in the stage of differentiation, from undifferentiated seminoma to more differentiated non-seminomas. In our studies, we aimed to characterize specific epigenetic features of GCT types that enable transcription derepression of the human endogenous retrovirus ERVWE1 in these tumors. We detected upregulated mRNA expression of TET1-3 dioxygenases in GCTs, especially of TET1 in seminomas. Moreover, seminomas showed low global levels of 5mC and 5hmC. TET1 knock-down in a seminoma-derived cell line resulted in a decreased amount of 5hmC and unchanged 5mC level. These results stress the dynamics of cytosine modifications, which has not been precisely described yet. Further, we observed high level of ERVWE1 transcript together with efficient RNA splicing in seminomas. Detected ERVWE1 transcription is independent of the expression of other examined endogenous retroviruses. ERVWE1 transcription derepression corresponds with the low global level of 5mC detected in seminomas, which involves extensive DNA hypomethylation of the ERVWE1 promoter. We propose the high TET1 dioxygenase expression as s marker of undifferentiated GCTs. Furthermore, we...
|
|
|
Some molecular mechanisms of calcium phosphate metabolism in diabetes mellitus
Vedralová, Marcela ; Černá, Marie (advisor) ; Kalvodová, Bohdana (referee) ; Brunerová, Ludmila (referee)
We chose to study polymorphisms of vitamin D receptor gene (VDR), parathyroid hormone gene (PTH) and CASR gene whose protein products significantly affect calcium phosphate metabolism and are implicated in the pathogenesis of diabetes, which may also involve kidney damage. The aim of this study was to find out whether the VDR, PTH and CASR polymorphisms are associated with the risk of developing diabetes mellitus and its renal complications. The frequencies of alleles and genotypes were studied in the groups of diabetic patients a with and without kidney complications using either TaqMan probes or PCR-RFLP. Comparison of patients with diabetic nephropathy (DN) and healthy subjects identified statistically significant difference for the FokI polymorphism in VDR gene (P<10-4) and also for the BstBI polymorphism in PTH gene (P=0,023). DraII polymorphism in PTH gene is associated with the risk of developing diabetes and its renal complications. In DN patients, the BBFFAATt haplotype of VDR gene was more frequent than in healthy subjects (P=0,046), and the BbFFAaTt variant was more frequent than in DM2 patients (P=0,018). The BBDD haplotype of PTH gene seems to be a predisposing factor for diabetes itself (P=0,019). The allele frequencies and genotype distribution for rs3804594 and rs1042636...
|
guest ::
RSS feed.