|
|
Interrelationship of basic biochemical parameters in patients of type II diabetes mellitus
Mazurová, Monika ; Boušová, Iva (referee) ; Mladěnka, Přemysl (advisor)
Diabetes mellitus 2nd tipe is heavy and very frequent civilizing disease in our population. The disease is characterized by hyperglycemia and other metabolism abnormalities are prezent. Assesment of glycemia in plasma (serum) is used for a diagnostics of the disease. Indicator of long- term compensation of diabetes is glycosylated haemoglobin. There were monitored possible dependensies between biochemical indicators in this work that are assesmented to diabetic patiens commonly. The dates were analyse from 54 diabetic patiens between the ages of 40 to 82 who visit a diabetic clinic periodically. The samples were analyzed by Cobas Integra 800 analyzer by Roche (Germany). Glucose in serum and in urine was analyzed by enzymatic metod with hexocinase and glycosylated haemoglobin was analyzed by imunochemical metod based on thurbidimetric inhibition in haemolysed blood. Paerson's correlative coefficient was used for appreciation of dependencies between variables. The highest correlative coefficient was found between glucose measuring in morning urine and glucosic losses by urine for 24 hours. Statistically important correlation were also found between glucose in serum and in urine or glucosic losses by urine for 24 hours and between glucose in serum and glycosylated haemoglobin. Because of relatively...
|
|
|
Optimization of Electrophoretic Methods for Study of Glycation of Aspartate Aminotransferase by Methylglyoxal
Bacílková, Eliška ; Szotáková, Barbora (referee) ; Boušová, Iva (advisor)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Mgr. Eliška Bacílková Supervisor: PharmDr. Iva Boušová, Ph.D. Title of thesis: Optimization of electrophoretic methods for study of glycation of aspartate aminotransferase by methylglyoxal Main scope of this thesis was the optimization of several electrophoretic methods for monitoring of the course of glycation of aspartate aminotransferase (AST) by methylglyoxal as well as of the effect of selected antioxidants at this process. I observed formation of high-molecular protein aggregates using denaturing electrophoresis (SDS- PAGE) and Western blotting and changes in the molecular charge of AST during glycation using native PAGE. In both types of electrophoresis, I searched for suitable concentration of the separation gel and the amount of loaded protein. Further, I compared three routinely used staining methods (Silver Staining, Coomassie Blue R-250 and G-250). I found out that the high-molecular aggregates with molecular weight about 130 and 160 kDa, which corresponds to tri- and tetramers of AST, are formed with increasing concentration of methylglyoxal. Using immunochemical detection, I detected glycation products with molecular weight mainly about 70 and 130 kDa and with molecular...
|
|
|
OCT activity and glycation damage in rat liver mitochondria
Merhautová, Lucie ; Dršata, Jaroslav (advisor) ; Boušová, Iva (referee)
OCT ACTIVITY AND GLYCATION DAMAGE IN RAT LIVER MITOCHONDRIA Lucie Merhutová Diploma thesis. Charles University in Prague, Faculty of Pharmacy in Hradec Králové. 2007 This work was created following investigations carried out in LBBCV (Laboratoire de Biologie et Biochimie Cellulaire du Vieillissement) at University Paris Diderot - Paris 7. The team of this laboratory studies the mechanisms of posttranslational non-enzymatic modifications of proteins, involved in the pathophysiology of aging. Glycation is one of the mechanisms responsible for the modification of intracellular macromolecules leading to the loss of their structure and function. In previous investigations, ornithine carbamoyltransferase (OCT), the second enzyme of urea cycle, has been identified as one of the markedly glycated proteins in liver mitochondria of senescent rats. The goal of this work was to investigate in vitro and ex vivo glycation of OCT and the effect of glycation on its function. Purified OCT and mitochondrial matrix extracts were incubated with methylglyoxal, one of the most reactive physiological glycating agents, and submitted to the enzymatic assay and immunochemical analysis. We demonstrated that methylglyoxal modifies OCT both in vitro and ex vivo and causes rapid and extensive decrease in its enzymatic activity....
|
|
|
The effects of natural antioxidants in the model of protein glycation
Švehlová, Klára ; Dršata, Jaroslav (advisor) ; Boušová, Iva (referee)
The effects of natural antioxidants in the model of protein glycation In my diploma thesis, I was concerned with direct and indirect effects of antioxidants of natural origin (hydroxycinnamic acids, HCAs) on the catalytic activity of aspartate aminotransferase as well as structure-activity relationship of individual HCAs. I used new model of methylglyoxal-induced glycation of aspartate aminotransferase, which has not been tested at the Department of Biochemical Science, Faculty of Pharmacy so far. I observed the structure-activity relationship of selected HCAs. Their antiglycation effect increased with the increasing number of hydroxyl groups. The methylglyoxal model of glycation proved to be suitable for farther research of other compounds. Further, I observed the interactions between HCAs and the molecule of enzyme itself using absorption and fluorescence spectroscopy. Those interactions were apparent at the acquired spectra. HCAs bind most probably only to the protein part of the enzyme and not to the coenzyme. The effect of all HCAs was concentration-dependent.
|
|
|
Study of antioxidant and cytoprotective effects of intracellular iron chelators in catecholamine-induced cardiomyocyte injury
Koubková, Lucie ; Šimůnek, Tomáš (advisor) ; Boušová, Iva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Kralove Department of Biochemical Sciences Candidate: Lucie Koubková Supervisor: Profa . Dra . Natércia Teixeira Profa . Dra . Georgina Correia da Silva Doc. Pharm.Dr. Tomáš Šimůnek, Ph.D. Title of diploma thesis: Evaluation of anti-aromatase activity of isoflavones from biotransformed soya Breast cancer belongs to the most frequently diagnosed cancers in women. Approximately 60% of breast cancer is hormone-dependent containing estrogen receptors and requiring estrogen for tumour growth. Estrogens are able to promote transcription of genes involved in cell proliferation. Aromatase is responsible for the final step of estrogen biosynthesis and its inhibition has been considered as an important target for the treatment of estrogen-dependent breast cancer. Soya is a source of phytoestrogens, such as genistein and daidzein, which are thought to be effective in breast cancer prevention. However, the mechanisms associated with this effect are not fully understood. In this study we evaluated the effects of a soya extract containing genistein and daidzein on aromatase activity. Our results demonstrated that this extract had the ability to inhibit aromatase in a dose-dependent manner in placental microsomes and also in the MCF-7aro breast cancer...
|
|
|
Influence of hydroxycinnamic acids on methylglyoxal-induced protein glycation
Vejvalková, Hana ; Boušová, Iva (advisor) ; Trnková, Lucie (referee)
Author: Hana Vejvalková Publishing place: Charles University in Prague, Faculty of Pharmacy in Hradec Králové Title: Influence of hydroxycinnamic acids on methylglyoxal-induced protein glycation Supervisor: PharmDr. Iva Boušová, Ph.D. The diploma thesis was aimed at potencial antiglycation effect of antioxidants of natural origin (hydroxycinnamic acids) on the modification of bovine serum albumine and asparate aminotransferase by methylglyoxal as a glycating agent. I monitored the effect of hydroxycinnamic acids on generation of glycation products using fluorescent, absorption, and electrophoretic methods. The influence of hydroxycinnamic acids on the glycation of model proteins was ambiguous. The only derivative with proven antiglycation effect on production of non-specific glycation products and pentosidin measured by fluorescence and spectrophotometry was p-coumaric acid in 100 μM concentration. On the other hand, chlorogenic acid 100 μM displayed strong proglycation effects in all methods used for detection of glycation products. The effect of all tested substances was concentration dependent. Results show that the more substituents given acid contained in its molecule, the stronger proglycation effect this compound exerted.
|
|
|
Formation of carbonyl compounds by non-enzymatic protein glycation
Krautová, Lenka ; Boušová, Iva (advisor) ; Dršata, Jaroslav (referee)
Author: Lenka Krautová Publishing place: Charles University in Prague, Faculty of Pharmacy in Hradec Králové Title: Formation of carbonyl compounds by non-enzymatic protein glycation Supervisor: PharmDr. Iva Boušová, Ph.D. This diploma thesis was aimed at the optimization of carbonyl groups determination in biological material. Carbonyl groups are formed by effect of protein oxidation or glycation. As a model protein, I used bovine serum albumin for spectrophotometric and aspartate aminotransferase for electrophoretic method of assessment, which were glycated by several glycating agents (glucose; glucose + copper; fructose; fructose + copper; methylglyoxal). I optimized different parameters of spectrophotometric determination: solvent, concentration of reagent, amount of protein, and duration of incubation. I managed to find suitable conditions of this determination, which I have used in subsequent experiments with different glycating agents. At electrophoretic determination, I performed SDS-PAGE with following silver staining or immunoblotting with chemiluminescent detection. The results of spectrophotometric determination showed that methylglyoxal has the highest glycating ability, followed by fructose with addition of cupric ions, fructose itself, glucose with addition of cupric ions, and the...
|
|
| |
|
|
Effect of flavonoids on the metabolism of xenobiotics
Smetanová, Anna ; Szotáková, Barbora (advisor) ; Boušová, Iva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Title, Name, Surname of candidate: Mgr. Anna Smetanová Title, Name, Surname of tutor: Doc. Ing. Barbora Szotáková, Ph.D. Title of a rigorous work: The influence of flavonoids on the metabolism of xenobiotics Flavonoids are wide spread plant secondary metabolites with the protective effect against the cancer. One of the protective mechanisms of action is the modulation of the biotransformation enzymes. Xenobiotics - heterocyclic aromatic amines are presumably carcinogenic compounds generated during the ordinary cooking of meat and fish. The first metabolic step of the heterocyclic aromatic amines is the procarcinogen activation to the active N-hydroxy metabolites by cytochrome P4501A2 (CYP1A2). It would be desirable if the flavonoids in the appropriate manner modulate the enzymatic activity to eliminate the undesirable effects of the heterocyclic aromatic amines. The aim of this study was to investigate the influence of the rutin and quercetin flavonoids and heterocyclic aromatic amines 2-amino-3-methylimidazo[4,5- f]quinoline (IQ) and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) on CYP1A activity in the cell line LS174T and HCT-8. Rutin, quercetin, IQ and MeIQx were not toxic for cells up...
|
|
|
Natural compounds with potential antioxidant activity; in vitro study in a model of protein glycoxidation
Boušová, Iva ; Dršata, Jaroslav (advisor) ; Stoklasová, Alena (referee) ; Patočka, Jiří (referee)
1. SUMMARY IN CZECH (ABSTRACT) Reaktivní formy kyslíku a dusíku (RONS) a volné radikály se staly v posledním desetiletí významným předmětem studia v mnoha oborech včetně medicínských. Stav, při němž je rovnováha mezi vznikem a odbouráváním volných radikálů v organismu posunuta ve prospěch jejich vzniku, se nazývá oxidační stres. Při nadbytku volných radikálů v organismu dochází k poškození biomolekul a tkání, což může vést až ke vzniku onemocnění. Oxidační stres se podílí na vzniku celé řady onemocnění, např. diabetu mellitu, aterosklerózy, revmatoidní artritidy, Alzheimerovy choroby a dalších, ale také na fyziologickém procesu stárnutí. Ve snaze pozastavit průběh stárnutí a odvrátit vznik onemocnění se začala používat různá potravní aditiva s obsahem vitamínů a antioxidantů přírodního původu. Diabetes mellitus je onemocnění charakterizované hyperglykémií a je vždy provázeno oxidačním stresem. Glukóza a různé intracelulární cukry (fruktóza, ribóza, glyceraldehyd a další) se kovalentně váží na volné aminoskupiny proteinů a způsobují poškození jejich struktury. Tento proces se nazývá neenzymová glykace. Sledem následných reakcí vznikají pozdní produkty pokročilé glykace (AGEs), které se podílí na tvorbě kovalentních vazeb mezi sousedními molekulami proteinů (crosslinks). Mezi nejvýznamnější AGEs patří N--...
|
guest ::
