|
|
Examining the significance of peptides regulating the intake of food and the nutritional state of children and adolescents
Bronský, Jiří ; Průša, Richard (advisor) ; Pelouch, Václav (referee) ; Hyánek, Josef (referee)
Introduction: Regulation of food intake and nutritional status is mediated by complex interactions of regulatory peptides of the central nervous system, gastrointestinal tract and adipose tissue. These systems are connected by feedback loops which inform the centre about amount of ingested food and energy reserves in the organism. Dysfunction of any of these regulatory areas may lead to changes in nutritional status of the organism. Methods: We used radioimmunoassay to measure plasma levels of orexin A, total ghrelin and serum levels of leptin and enzyme immunoassay to measure serum levels of adiponectin in healthy subjects and in children with obesity, anorexia nervosa, Crohn's disease and celiac disease and we evaluated the influence of nutritional therapy on these levels. Moreover, we evaluated relationship of these regulatory peptides to other biochemical and anthropometrical factors of nutritional status. We also measured plasma levels of total and unreduced amylin by enzyme immunoassay with immunofluorescence detection in adult patients with osteoporosis, type II diabetes mellitus and in the control group. Results: During reduction of body weight in obese children and adolescents, there were statistically significant changes of plasma orexin A levels and total ghrelin levels, but we haven't seen any...
|
|
|
Study of mitochondrial ultrastructure and functions in selected mitochondrial and lysosomal storage disorders
Kostková, Olga ; Hansíková, Hana (advisor) ; Šmíd, František (referee) ; Hyánek, Josef (referee)
This thesis has been worked out in The laboratory for study of mitochondrial disorders (Departement of Pediatrics, 1st Faculty of Medcine, Chales university in Prague) and in cooperation with The Institute of Inherited Metabolic Disorders. Mitochondrial disorders represent a heterogeneous group of diseases with the onset at any age from neonatal period till adulthood, mostly presented with very severe clinical courses of disease. The mammalian organism is fully dependent on mitochondrial oxidative phosphorylation system as on the major energy producer of the cell. Therefore the mitochondrial disorders affect mainly high energy demanded tissues such as brain, heart or muscle. Simillar phenotype is observed in many lysosomal storage disorders. Despite of expanding knowledge of molecular basis of mitochondrial and lysosomal disorders, it may be still difficult to explain the exact pathogenesis of disease as well as the prognosis for patients and their families. Mitochondrial functions affect more than just energy production; they contribute in initiation of apoptosis, in cellular calcium homeostasis, and in production of reactive oxygene species. Disturbed mitochondria become a goal of autophagy mediated by the lysosomal compartement. The results of our study enable: 1. better understanding of the tissue...
|
|
|
název v anglickém jazyce není uveden
Belošovičová, Hana ; Calda, Pavel (advisor) ; Hyánek, Josef (referee) ; Kvasnička, Jan (referee)
Souhrn Cíl: Stanovit hladiny celkového plazmatického cysteinu, homocysteinu, cysteinylglycinu a glutathionu u normotenzních primigravid ve druhém a třetím trimestru a porovnat je s hladinami u těhotných s následným rozvojem preeklampsie. Metodika: Do studie bylo zařazeno 65 zdravých primipar. U každé těhotené byl nabrán vzorek krve v 19.-21. a 30.-32. týdnu těhotenství. Hladiny celkového plazmatického cysteinu, homocysteinu, cysteinylglycinu a glutathionu byly stanoveny HPLC metodou. Hladiny aminothiolů v druhém a třetím trimestru byly porovnány pomocí párového t testu. Hladiny cysteinu u těhotných byly porovnány s hladinami cysteinu u kontrolního souboru netěhotných žen. Výsledky: Hladina celkového plazmatického cysteinu u normotenzních těhotných byla signifikantně nižší ve třetím trimestru v porovnání strimestrem druhým (medián 176.1 |amol/l [163.0, 189.4] versus 187.4 |amol/l [178.7, 205.2], P < 0.001). Hladiny ostatních aminothiolů se nelišily. U třech z 65 těhotných došlo k rozvoji mírné preeklampsie. Hodnoty t-Cys ve třetím trimestru u těhotných s následným rozvojem preeklampsie vykazovaly nárůst nebo menší pokles než u normotenzních těhotných. Závěr: Celkový plazmatický cystein je signifikantně nižší ve třetím trimestru než v trimestru druhém. Tento pokles je pravděpodobně dán spotřebou cysteinu...
|
|
| |
|
|
Asymmetric Dimethylarginine and Its Relation to Atherogenesis
Široká, Romana ; Racek, Jaroslav (advisor) ; Teplan, Vladimír (referee) ; Hyánek, Josef (referee)
ASYMMETRIC DIMETHYLARGININE - COMPARSION OF CHROMATOGRAPHY AND IMMUNOMETRIC METHODS Objective:Asymmetric dimethylarginine (ADMA) is often discussed in connection with hyperhomocysteinemia and its toxic effect on vessel wall. ADMA concentration is usually measured by HPLC (High Performance Liquid Chromatography) after previous derivatisation. Recently, ELISA (Enzym Linked Immuno Assay) methods for ADMA determination were introduced and ELISA kits are commercially available. Method and Result:The aim of the study was to compare HPLC and ELISA methods for ADMA determination. For HPLC determination we used equipments from Thermo separation product (Florida, USA). After solid-phase extraction on polymer cation-exchange column and the following derivatisation with o-phthaldialdehyde the samples were separated using C18 column (mobile phase 8.7% acetonitril, 50 mmol/l phosphate buffer, pH 6.5) and a fluorescence detector. NG-monomethyl-L-arginine was used as an internal standard. ADMA® ELISA kit, based on a competitive principle, was obtained from DLD Diagnostika, Hamburg, Germany. ADMA was measured in EDTA plasma of 40 healthy blood donors and 40 hemodialysis patients with hyperhomocysteinemia. Conclusion: In spite of different principles both methods showed a very good correlation (r = 0.944, p<0.0001). ELISA...
|
|
|
Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies
Fornůsková, Daniela ; Zeman, Jiří (advisor) ; Hyánek, Josef (referee) ; Stiborová, Marie (referee)
Mgr. Daniela Fornuskova PhD thesis Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies ABSTRACT The mammalian organism fully depends on the oxidative phosphorylation system (OXPHOS) as the major energy (ATP) producer of the cell. Disturbances of OXPHOS may be caused by mutations in either mitochondrial DNA (mtDNA) or nuclear DNA (nDNA). One part of the thesis is focused on the role of early and late assembled nuclear-encoded structural subunits of cytochrome c oxidase (CcO) as well as Oxa1l, the human homologue of the yeast mitochondrial Oxa1 translocase, in the biogenesis and function of the human CcO complex using stable RNA interference of COX4, COX5A, COX6A1 and OXA1L, as well as expression of epitope-tagged Cox6a, Cox7a and Cox7b, in HEK (human embryonic kidney)- 293 cells. Our results indicate that, whereas nuclear- encoded CcO subunits Cox4 and Cox5a are required for the assembly of the functional CcO complex, the Cox6a subunit is required for the overall stability of the holoenzyme. In OXA1L knockdown HEK-293 cells, intriguingly, CcO activity and holoenzyme content were unaffected, although the inactivation of OXA1 in yeast was shown to cause complete absence of CcO activity. In addition, we compared OXPHOS protein deficiency patterns in mitochondria from skeletal...
|
|
|
Advanced lipidomics in selected clinical conditions
Staňková, Barbora ; Tvrzická, Eva (advisor) ; Hyánek, Josef (referee) ; Kazdová, Ludmila (referee)
Abnormalities of lipid metabolism are considered risk factors for cardiovascular, metabolic, nephrologic diseases amd some malignancies, as well. Nowadays, a lot of effort is devoted to study new risk factors and surrogate markers of conditions mentioned above to improve their prognosis and decrease mortality. The aim of this thesis was to provide a comprehensive survey of lipid metabolism, characteristics of different lipid compounds in health and diseases and of possibilities of utilization of selected lipid parameters in the diagnostics of pathological conditions listed above. Selected lioid parameters were observed in several studies, focused on specific pathological conditions. Besides conventional lipid analytes, the composition of fatty acids in plasma lipid pools was studied in healthy controls, and in the patients suffering from metabolic syndrome, chronic pancreatitis, and pancreatic cancer, as well. Markers of an oxidative stress (oxidatively modified LDL particles and conjugated dienes in precipitated LDL) were assessed in healthy controls, patients with metabolic syndrome, chronic pancreatitis, pancreatic cancer, and in the patients with different concentrations of plasma apoB-48, too. LDL particles subfraction were investigated in healthy controls, in the patients with different...
|
|
| |
|
|
Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies
Fornůsková, Daniela ; Zeman, Jiří (advisor) ; Hyánek, Josef (referee) ; Stiborová, Marie (referee)
Mgr. Daniela Fornuskova PhD thesis Biochemical and molecular studies of cytochrome c oxidase and ATP synthase deficiencies ABSTRACT The mammalian organism fully depends on the oxidative phosphorylation system (OXPHOS) as the major energy (ATP) producer of the cell. Disturbances of OXPHOS may be caused by mutations in either mitochondrial DNA (mtDNA) or nuclear DNA (nDNA). One part of the thesis is focused on the role of early and late assembled nuclear-encoded structural subunits of cytochrome c oxidase (CcO) as well as Oxa1l, the human homologue of the yeast mitochondrial Oxa1 translocase, in the biogenesis and function of the human CcO complex using stable RNA interference of COX4, COX5A, COX6A1 and OXA1L, as well as expression of epitope-tagged Cox6a, Cox7a and Cox7b, in HEK (human embryonic kidney)- 293 cells. Our results indicate that, whereas nuclear- encoded CcO subunits Cox4 and Cox5a are required for the assembly of the functional CcO complex, the Cox6a subunit is required for the overall stability of the holoenzyme. In OXA1L knockdown HEK-293 cells, intriguingly, CcO activity and holoenzyme content were unaffected, although the inactivation of OXA1 in yeast was shown to cause complete absence of CcO activity. In addition, we compared OXPHOS protein deficiency patterns in mitochondria from skeletal...
|
|
|
název v anglickém jazyce není uveden
Belošovičová, Hana ; Calda, Pavel (advisor) ; Hyánek, Josef (referee) ; Kvasnička, Jan (referee)
Souhrn Cíl: Stanovit hladiny celkového plazmatického cysteinu, homocysteinu, cysteinylglycinu a glutathionu u normotenzních primigravid ve druhém a třetím trimestru a porovnat je s hladinami u těhotných s následným rozvojem preeklampsie. Metodika: Do studie bylo zařazeno 65 zdravých primipar. U každé těhotené byl nabrán vzorek krve v 19.-21. a 30.-32. týdnu těhotenství. Hladiny celkového plazmatického cysteinu, homocysteinu, cysteinylglycinu a glutathionu byly stanoveny HPLC metodou. Hladiny aminothiolů v druhém a třetím trimestru byly porovnány pomocí párového t testu. Hladiny cysteinu u těhotných byly porovnány s hladinami cysteinu u kontrolního souboru netěhotných žen. Výsledky: Hladina celkového plazmatického cysteinu u normotenzních těhotných byla signifikantně nižší ve třetím trimestru v porovnání strimestrem druhým (medián 176.1 |amol/l [163.0, 189.4] versus 187.4 |amol/l [178.7, 205.2], P < 0.001). Hladiny ostatních aminothiolů se nelišily. U třech z 65 těhotných došlo k rozvoji mírné preeklampsie. Hodnoty t-Cys ve třetím trimestru u těhotných s následným rozvojem preeklampsie vykazovaly nárůst nebo menší pokles než u normotenzních těhotných. Závěr: Celkový plazmatický cystein je signifikantně nižší ve třetím trimestru než v trimestru druhém. Tento pokles je pravděpodobně dán spotřebou cysteinu...
|
guest ::
