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The role of orexins in food intake and chosen pathological phenomena
Zelená, Anna ; Sumová, Alena (advisor) ; Balounová, Kateřina (referee)
Orexins are neuropeptides produced in the lateral hypothalamus in the brain and peripheral organs. These neuropeptides are essential in regulating arousal and energy balance. They also participate in the reward system and secure water homeostasis. Orexins bind to the orexin receptors differently expressed in various tissues. The binding of orexins to their receptors couples with downstream signalling pathways, e.g., Ca2+ , diacylglycerol and protein kinase C pathways. Low concentration of adenosine triphosphate leads to the inactivation of orexin-expressing neurons and the reduction of energy expenditure. On the contrary, high intracellular concertation of adenosine triphosphate activates energy expenditure. Food intake is regulated by orexins and glucose, sirtuin 1, forkhead box transcription factor 2, and leptin. This thesis aims to find connections between the role of orexins in regulating food intake and the development of some pathological phenomena, e.g., hypertension, obesity, and anorexia nervosa. Keywords orexin, obesity, anorexia nervosa, glucose, insulin, stress
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The development of reliable method for glucosylation and lactosylation of sphingolipids
Hošek, Adam ; Opálka, Lukáš (advisor) ; Karabanovich, Galina (referee)
Title of the master thesis: The development of reliable method for glucosylation and lactosylation of sphingolipids Candidate: Adam Hošek Supervisor: PharmDr. Lukáš Opálka, Ph.D. Consultant: Dr. Georgios Paraskevopoulos, Ph.D. Charles University, Faculty of Pharmacy in Hradec Králové Skin Barrier Research Group, Department of Organic and Bioorganic Chemistry Ceramides are hydrophobic sphingolipids consisting of a sphingoid base and an acyl chain. They are found in the stratum corneum, the uppermost layer of the epidermis, where they are important part of the skin barrier, but they can also be found in brain tissue and inside human cells. Glucosylceramides are the simplest glycosphingolipids, and they contain a single glucose molecule in their structure. They are found in all types of human cells, where they not only serve as precursors for the biosynthesis of other, more complex glycosphingolipids, but also have their own physiological functions. The most significant is their influence on cell growth, development, differentiation and apoptosis. Disruptions in their synthesis or degradation can lead to serious health problems and are also a risk factor for the future development of a wide range of diseases. Their total absence in the body is incompatible with life. A more thorough study of their...
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On-line derivatization of saccharides in capillary electrophoresis
Mareš, Vít ; Křížek, Tomáš (advisor) ; Coufal, Pavel (referee)
Saccharides chains may suffer from various inaccuracies during their biosy- thesis, that effects their biologic functions. Therefore, they can be used for dia- gnosis of some diseases or quality control in biopharmaceutics. Capillary electro- phoresis has its place in diagnosing these occuring errors. In this thesis a method of on-line derivatization of saccharides in fused-silica capillary with 50 cm in lenght and 50 µm in inner diameter was developed. Gluco- se was used as a model molecule, which was labeled by a fluorescent derivatization reagent, 7-aminonaphthalene-1,3-disulfonic acid. Derivatization of glucose and its separation needed optimalization. First was determined an electrophoretic mo- bility of the derivatization reagent in 0.5; 1.0 and 2.5M acetic acid and 20mM phosphate buffer (pH = 3.5). Both, 2.5M acetic acid and 20mM phosphate buffer, were tested as solvents for 7-aminonaphthalene-1,3-disulfonic acid. The reaction with glucose was the best in acetic acid solution. On the contrary, phosphate buf- fer was found to be the best background electrolyte for the separation. Derivatized glucose was detected by UV detector at 229 nm. Henceforth, the reaction con- ditions were adjusted such as reaction time, voltage and time of its application. Input voltage was applied negative due to the...
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Determination of glucose by different analytical systems
ŠTECHEROVÁ, Monika
The determination of blood glucose by three different analytic systems - Abbott Architect c16000, glucometr SensoStra GL 30 Touch and the self-monitoring glucometer Contour Plus. overall 30 random patient samples were measured at four analyzers. The mesured values for the internal control quality were evaluated and the regular verifications of the methods were realized at the analyzer Abbott Architect c16000 and glucometers SensoStar GL 30 Touch.
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Comparison of the gold microelectrode and conventional electrode for the saccharide determination using pulsed amperometry
Smítková, Karolína ; Dejmková, Hana (advisor) ; Schwarzová, Karolina (referee)
This bachelor's thesis deals with combination of pulsed amperometric detection (PAD) with flow injection analysis (FIA) in order to use them to detect glucose and consequently carbohydrates in solution. Pulsed amperometry is efficient detection method for many of organic compounds in which, as the name suggests, a sequence of three or four potential pulses is applied at the working electrode. If the pulse parameters are suitable, then it is possible to eliminate the unwanted passivation of the working electrode due to the adsorption of oxidation products and intermediate products of aliphatic compounds, which would lead to a reducion of its active surface. The aim od this work is to show the effect of the change of the value of the applied potential pulses on the peak heights corresponding to the oxidation of the glucose in solution and to evaluate their repeatability. As a working electrode, it compares a gold electrode of conventional size and a gold microelectrode. The determination was performed in a borate buffer with pH = 10. First of all, cyclic voltammograms of solution with concentration of glucose 1·10-2 mol dm-3 were measured to obtain the initial values of the applied pulses. Subsequently, the dependence of the peak heights on their injection order for different values of individual...
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Double-gate biosensor of glucose based on functionalized graphene
Malatinová, Michaela ; Pavlásková, Lucie (referee) ; Bartošík, Miroslav (advisor)
V tejto bakalárskej práci bola skúmaná funkcionalizácia grafénu s sukcinimidyl ester pyrenbutanovou kyselinou a enzýmom glukóza oxidáza pre detekciu glukózy biosenzorom. Funkcionalizovaný grafén bol implementovaný ako kanál v poľom riadenom tranzistore pre výrobu biodetekčného zariadenia. Proces funkcionalizácie potvrdila Ramanova spektroskopia a mikroskopia atomárných síl. Charakterizácia senzoru a jeho kvalít bola sledovaná pomocou transferových kriviek a časovej odozvy senzoru. Posun Diracovho bodu a jeho poloha boli analyzované v závislosti na rôznych koncentráciach glukózy s ohľadom na efektivitu funkcionalizácie. Výber hradlovej elektródy v grafénovom poľom riadenom tranzistore bol skúmaný vzhľadom na jeho možný dopad na výsledky merania.
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The comparation of embryotoxical effect of insulin and glucose by the method CHEST.
Turková, Aneta ; Peterka, Miroslav (advisor) ; Likovský, Zbyněk (referee)
If gravid women suffer from diabetes,their unborn children have 10x higher risk of development of malformation,prenatal and postnatal death and post partum complications than children of women belonging to healthy population.The main and very controversial potential teratogenic factors are glucose and insulin.However,there are very ambivalent opinions on which one of these two substances causes damage to embryo. Therefore,the aim of this diploma thesis was to contribute to solving of this problem and test direct embryotoxicity of insulin and glucose. For solving of this issue, the so-called CHEST.The principle of this method is creation of a window in eggshell and consequent subgerminal or intraamnial application of the substance being tested.Embryos were tested from the second until the sixth incubation day.Firstly, two types of insulin were injected.Injected doses varied between 3µg/3µl to 0,003µg/3µl. Then glucose was tested,with dosage of 300µg/3µl and 30µg/3µl. Embryotoxic effect was detected for both types of insulin. The beginning of embryotoxicity line of insulin's lies between the dosage of 0,03µg/3µl and 0,003µg/3µl. From embryotoxic effect, death of embryos predominated over development of congenital malformations. Only after application on the second incubation day, there was increased...
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Utilization of energy substrates in white and brown adipose tissue in metabolic syndrome
Seďová, Karolína ; Škop, Vojtěch (advisor) ; Šilhavý, Jan (referee)
1 Abstrakt Hnědá tuková tkáň je unikátní termoregulační orgán, její funkcí je přeměňovat energii ve formě protonového gradientu na energii tepelnou a zároveň snižovat efektivitu metabolismu. Její funkce se zdá být nadějí v léčbě metabolického syndromu a komplikací s ním spojených. Efekt hnědé tukové tkáně v léčbě MS však stále není objasněn a proto jsme chirurgicky odstranili tuto tkáň u potkanů, kteří trpí metabolickým syndromem, abychom objasnili důsledky extirpace na parametry metabolického syndromu. V této práci jsme sledovali aktivitu interskapulární tukové tkáně na parametry metabolického syndromu. Srovnávali jsme parametry mezi kmeny HHTg potkanů a kontrolním kmenem potkanů Wistar a pozorovali jsme nižší hmotnost potkanů kmene HHTg přes projevy ostatních parametrů metabolického syndromu, jako je dyslipidémie, inzulinová rezistence tukové tkáně, vyšší hodnoty NEMK v krvi. Zjistili jsme, že tito potkani mají významně vyšší aktivitu iBAT, jak v inkorporaci palmitátu do lipidů, tak ve spalování lipidů ve formě CO2 a zejména v nižším věku. Tato zjištění nás vedla k hypotéze, že iBAT by mohla mít zásadní vliv na parametry MS u neobézních potkanů. Extirpací iBAT u mladých potkanů, ani u potkanů v pokročilejším věku však neměla zásadní vliv na metabolismus ani na hmotnost. Dokonce došlo ke zlepšení některých...
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