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Comparison of the determination of hormones (Follicle stimulating hormone, Luteinizing hormone, Prolactin, Testosterone, Progesteron) by two analytical systems. Converting accredited method and its verification.
Kucejová, Soňa ; Martínková, Markéta (advisor) ; Mrízová, Iveta (referee)
Analytical system ARCHITECT i2000SR was verified according to requirements of ÚLBLD VFN and 1. LF UK laboratory in Prague. Repeatability, intermediate precision, and measurement uncertainty were determined as performance parameters for verification of analytical assays for testosterone, progesterone, luteinizing hormone, follicule stimulating hormone and prolactin. Results of Lyphochek control samples, which were measured, were consistent with values given by manufacture. Repeatability: coefficients of variation for testosterone Lyphochek 1 6,81%, for Lyphochek 3 6,40%, progesterone 2,4% and 1,8%, luteinizing hormone 5,38% and 1,89%, follicle stimulating hormone 5,12% and 3,24% prolactin 1,45% a 1,83%. Intermediate precision: coefficients of variation for testosterone Lyphochek 1 6,02%, Lyphochek 2 3,60%, Lyphochek 3 3,07%, progesterone 7,9%, 4,9% and 5,8%, luteinizing hormone 4,50%, 5,51% and 5,83%, follicle stimulating hormone 4,00%, 3,72% and 4,87%, prolactin 4,60%, 4,20% and 5,00%. Measurement uncertainty: testosterone 6,02%, progesterone 7,9%, luteinizing hormone 5,83%, follicle stimulating hormone 4,87%, prolactin 5,00%. Analytical System Architect i2000SR was compared with previously used ADVIA Centaur system to find out, whether it is possible to convert the method Centaur Testosterone,...
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Determination of cortisol in saliva
Žánová, Magdalena ; Martínková, Markéta (advisor) ; Milichovský, Jan (referee)
This thesis deals with salivary cortisol levels measured by the ADVIA: Centaur CP Immunoassay System. Experimental saliva sampling was performed on 141 pro- bands from age 7 to 76. Volunteers were divided to groups according to their sex: males, females using hormonal contraceptives (HC) and females not using HC. Re- ference intervals of morning and evening salivary cortisol were defined: females not using HC 13,2 - 55,5 nmol/l and females using HC 15,5 - 44,2 nmol/l for mor- ning salivary cortisol. Females not using HC 4,0 - 16,6 nmol/l and females using HC 7,9 - 22,6 nmol/l for afternoon salivary cortisol. Males 15,8 - 47,7 nmol/l for morning salivary cortisol and 5,2 - 25,4 nmol/l for afternoon salivary cortisol. Dif- ferences in stated intervals were imperceptible in all monitored groups. However, it is necessary to maintain different reference intervals for both morning and evening sampling. Reference interval for morning sampling was 14,3 - 46,2 nmol/l and refe- rence interval for afternoon sampling was set at 4,0 - 22,2 nmol/l. Daily profiles of salivary cortisol were determined in 6 females and 4 males in four different parts of a day. The course of salivary cortisol levels corresponded in females not using HC, females using HC and males with circadian rhythm, which is subject to cortisol. Sa-...
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Laboratory tests for thyroid function assessment from the economic point of view
Stehlíková, Petra ; Martínková, Markéta (advisor) ; Hýsková, Veronika (referee)
In 2011 Recommendation for laboratory diagnostics of functional and autoimmune thyroid diseases was published. Its target is to recommend correct method when choosing thyroid examination parameters to general practitians and thus find a way how to save money on useless examinations which then can be used by hospital in other area. To verify physicians adhere to these recommendations file of VFN patients in Prague for year 2011 was used and statistically evaluated. Watched parameters included thyrotropin (TSH), free triiodothyronine (FT3), total triiodothyronine (T3), free thyroxine (FT4), total thyroxine (T4), thyroid peroxidase antibodies (anti - TPO), thyroglobulin antibodies (anti - Tg), thyroglobulin (Tg) and thyroxine - binding globulin (TBG). Basic test according to published recommendation should include only TSH and if out of reference limits then it should be followed by FT4 test or antidote assessment. Other parameters are unnecessary for initial examination, some of them are not used anymore or should be examined only by clinics specialized in endocrinology. Based on the file records many physicians are still ordering unnecessary examination parameters. In one year 13,8 % of unnecessary parameters out of 44 415 thyroid function examinations were determined. This is 1 095 013 points that could by...
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Laboratory Diagnosis of Primary Aldosteronism
Hulová, Dagmar ; Martínková, Markéta (advisor) ; Bárta, František (referee)
Primary aldosteronism is a common form of endocrine hypertension, characterized by inappropriate production of aldosterone by the adrenal cortex, usually because of the unilateral adenoma (APA) or bilateral hyperplasia (IHA). The aim of this study was answer the question, if it is possible to distinguish APA from IHA with the use of the postural or the confirmatory test. From the laboratory results of the postural test, it was found that for the differentiation between the two most common forms of primary hyperaldosteronism APA and IHA is the postural test unusable in most cases. By the confirmation test was confirmed that there is not suppression of aldosterone after the administration of the saline solution to patients with primary aldosteronism below 100 ng∙l-1 in most cases (for APA in 85 %, for IHA in 65 %), in the remaining cases the value of the aldosterone do not decrease below 50 ng∙l-1 A higher value of the aldosterone in APA after the administration of the saline solution in the confirmation test could be considered a laboratory indicator of overproduction in APA.
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Comparison of adsorption capacity and adsorption efficiency of diosmectite and charcoal on selected compounds that often cause acute intoxication in adult population of the Czech Republic
Mináriková, Michaela ; Martínková, Markéta (advisor) ; Mrízová, Iveta (referee)
In the treatment of acute intoxications, one of the treatment procedures is an antidote submission, e.g. diosmectite and activated charcoal, where an antidote is a substance which acts antagonistically and disturbingly with the toxic effect of a toxic substance. The aim of this work was to compare the adsorption capabilities of activated charcoal and diosmectite in selected model compounds which are the most common originators of acute intoxications in the adult population of the Czech Republic. The actual comparison of adsorption capabilities of these sorbents was preceded by issues search processing of ten groups of substances that cause the most acute intoxications and subsequent testing of the proposed method for future detailed testing of adsorption and adsorption efficiency of different sorbents. Of the ten groups of substances five model compounds were selected: dosulepin, acetylsalicylic acid, ibuprofen, promethazine and phenobarbital, on which adsorption of diosmectite, activated charcoal, and mixture of these sorbents was compared in the experimental part of this work. This comparison of the adsorption capabilities of sorbents was carried out not only under neutral conditions, but also in an alkaline and acidic environment which simulated physiological conditions in different parts of the...
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Protein kinase C ε and δ in chronically hypoxic heart
Kožichová, Kristýna ; Martínková, Markéta (referee)
1 Abstract Adaptation to chronic hypoxia confers long-term cardioprotective effects. We have shown, that protein kinase C (PKC) is involved in this cardioprotective phenotype. It has not been elucidated, which PKC isoform plays a role, but the most likely candidates are PKC ε a PKC δ. Therefore, the aim of this study was to analyze the expression and localization of PKC ε and PKC δ after the adaptation to chronic hypoxia. We have shown that adaptation to chronic hypoxia caused the activation and tranlocation of PKC δ to the mitochondrial and sarcolemmal membranes. Our study suggests that PKC δ plays an important role in the mechanism of cardioprotection induced by chronic hypoxia.
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Preparation and characterization of a bacterial protein YddV, a globin-coupled oxygen sensor diguanylate cyclase
Křížová, Věra ; Martínková, Markéta (advisor) ; Man, Petr (referee)
Heme-containing proteins are very important for proper function of various organisms, both prokaryotic and eukaryotic. The most well-known ones are myoglobin, hemoglobin and various peroxidases and cytochromes. The group of heme-containing proteins was recently extended by a new type of proteins, which are called heme-containing sensor proteins. These proteins consist of two domains, sensor domain, which is able to detect a signal (either a molecule of heme or a molecule of gas), and functional domain, which is often an enzyme or a transcriptional factor. Activity of the functional domain depends on the concentration of signal molecule, which binds to the sensor domain, which then causes change of its structure. Consequently, change of structure of the sensor domain also leads to change of structure of the functional domain. Heme-containing sensor proteins can be further divided into two smaller groups: heme sensor proteins and gas sensor proteins. Heme sensor proteins share a lot of features, for example binding of heme to a thiolate residue from cysteine, CP motif, etc. Gas sensor proteins detect a molecule of gas, especially oxygen, carbon monoxide and nitrogen monoxide. There have already been conducted various studies focusing on heme-containing sensor proteins, however, the mechanism of signal...
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Important roles of heme as a signal and a gas-sensing site: heme-sensing and gas-sensing proteins
Fojtíková, Veronika ; Martínková, Markéta (advisor) ; Hudeček, Jiří (referee)
Heme-containing sensor proteins are heme proteins, which are divided into two groups: heme-sensing and gas-sensing proteins. The function of heme-sensing proteins is affected by heme availability. Association (or dissociation) of heme moiety of heme-sensing protein regulates various physiological functions, including protein kinase activity, transcription and other important functions essential for cell survival. In gas-sensing proteins, heme acts as the sensing site for binding of gaseous molecules (including O2, NO and CO) and indirectly regulates physiological functions, including protein kinase activity, transcription and other important functions essential for cell survival. The recent studies on heme-containing senzor proteins published in scientific journals are summarized in this thesis. The experimental part of this thesis focused on the specific heme-containing sensor protein - a globin-coupled histidine kinase from Anaeromyxobacter sp. strain Fw 109-5 (AfGcHK). The aim of this thesis was to amplified and isolate plasmid carrying gen for AfGcHK. Consequently the protein was expressed in E.coli BL-21(DE3) and the protein was isolated. Based on the results, the isolation process was optimized. Moreover, the purified preparation of isolated AfGcHK was prepared in more than 99% of homogeneity....
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Assessment of laboratory tests efficiency in case of exocrine pancreas function determination
Šubčíková, Lenka ; Martínková, Markéta (advisor) ; Milichovský, Jan (referee)
Chronic pancreatitis is complex disease with complicated diagnosis. Nowadays there are the diagnosis and classification of chronic pancreatitis primarily based on imaging methods. In this study the results of two indirect tests of pancreatic exocrine function with different arrangement were compared. The pancreatic fecal elastase 1 was determinate by immunoassay with monoclonal antibody, as a simple screening test. The dynamics and kinetics of pancreatic exocrine secretion was observed by a breath test with 13 C-labeled substrate. The group of four volunteers was monitored for six months and analyzed each month. The mutual variability of these tests and their correlation was studied. The patients' (suspected suffering by chronic pancreatitis) anonymous data for these tests created by the Institute of laboratory biochemistry and laboratory diagnostics between 1999-2012 were statistical processed. We found, that the indirect tests of pancreatic exocrine function and determination of pancreatic enzymes does not correlate. Both of these tests have the specific diagnostic value mainly for diagnosis of pancreatic exocrine insufficiency. 1
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Study of mechanism of fibrin network formation
Božíková, Paulína ; Martínková, Markéta (advisor) ; Hýsková, Veronika (referee)
Serine protease thrombin plays an important role in the process of fibrin network formation by converting fibrinogen into fibrin monomer which spontaneously polymerizes to form fibrin network. The aim of this work was to characterize interactions between thrombin and surface adsorbed fibrin(ogen) or fibrin network to which thrombin can bind and initiate grow of the fibrin network. Activity of thrombin bound on fibrinogen or fibrin was determined spectrophotometrically in a relation to cleaved chromogenic substrate. Using the method of surface plasmon resonance fibrin network formation initiated by thrombin bound to fibrinogen or fibrin was observed. These networks were also visualized by atomic force microscopy. Determined value of affinity constant KD for interaction of fibrinogen in solution with a fibrin network prepared on surface is in agreement with previous experiments in which KD was determined from interaction of surface covalently bound fibrinogen with fibrin monomers in solution.
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