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Application of SPE and technology of monolithic columns for HPLC analysis of biologically active compounds.
Horčičková, Šárka ; Šatínský, Dalibor (referee) ; Solich, Petr (advisor)
The solid phase extraction (SPE) is an effective, reliable and selective modern method for sample preparation of biological material before chromatographic analysis. In this graduation thesis the new SPE method for isolation of vitamin D2 (ergocalciferol), D3 (cholecalciferol) and its metabolite 25 - (OH) D3 (calcidiol) simultaneously with vitamin A (retinol) and E (α-tocopherol) in human serum was developed. The vitamins were subsequently determined by validated high performance liquid chromatography method (HPLC) using tocol as internal standard. The developed SPE procedure: Firstly 250 l of human serum was pipetted to glass tube and 1000 l of cool ethanol (4řC) was added. Sample was shaken and left 10 minutes in a refrigerator at 4řC for deproteinisation. After centrifugation (4 000 x g, 15 minutes, 4řC) all supernatant was carefully removed and applied into pre-treatment SPE column (firstly washed with 1 ml of methanol, then with 1 ml of distilled water). During these two steps the sorbent must not be dried up. The vitamins were eluted by 1.5 ml of methanol and then 2.0 ml of n-hexane. The organic solutions were evaporated in the vacuum concentrator at a temperature of 45 ř C. The residue was dissolved in 250 l of methanol and analyzed by using HPLC method. HPLC method: Mobile phase - A:...
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HER2/neu and other markers in breast tumours.
Celbová, Lenka ; Doležel, Martin (referee) ; Hochmann, Jiří (advisor)
2. SUMMARY Author: Bc. Lenka Celbová Tutor: MUDr. Jiří Hochmann, CSc. Title of a work: HER-2/neu and other markers in breast tumours Diploma work Department of Biological and Medical Sciences Charles University in Prague, Faculty of Pharmacy in Hradec Králové Study programme: Healthcare bioanalytics The aim of our diploma work is to bring an overview of the tumour marker HER- 2/neu especially with focus on its activation, laboratory detection, relation to hormonal receptors, its role in tumorgenesis of breast carcinoma and also on the examination of its prognostic importace and its relation to the breast cancer treatment. In the clinical part of this review work we target especially the prognostic and predictive value of HER-2/neu. In many studies, a bad prognosis of HER-2/neu positive breast cancer was confirmed. In this connection the importance of determination of extracellular domain HER-2/neu in serum was examined in the literature. It is proved, that observation of this parameter is useful for monitoring of the patients with HER-2/neu positive carcinomas and there is also proved its predictive importance for the trastuzumab treatment. Another topic investigated in this work is expression of HER-2/neu de novo. Two studies, we found, confirm the opinion, that an amplification of HER-2/neu de novo can...
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In vitro cultures of medicinal of plants - VIII
Špandelová, Veronika ; Martin, Jan (referee) ; Tůmová, Lenka (advisor)
In vitro cultures of medicinal of plants - VIII Method of elicitation is one of the possibilities how to increase production of secondary metabolites in cultures in vitro. The effect of abiotic elicitor (3-jodo-4-methylfenyl)amide 5- terc-butyl-6-chloropyrazine-2-karboxylic acid in three concentrations on the flavonolignans and flavonoid taxifolin production in Silybum marianum callus and suspension culture was tested in this study. Elicitation was run in this time intervals: 6, 12, 24, 48, 72 and 168 hours. Cultivation proceeded on Murashige - Skoog medium with the addition of 10 mg/l of α- naphtylacetic acid. The content of flavonolignans and flavonoid taxifolin was determined by HPLC. The maximum content of flavonolignans (0,05%) in callus culture was found after 168 hours of elicitor treatment in concentration of c2 = 2,33 x 10-4 mol/l. The maximum production of silychristin (0,04%) was found after 6 hours of affecting elicitor c1 = 2,33 x 10-3 mol/l. There wasn't statistical significant increasing of flavonolignans and taxifolin production in suspension culture. The maximum production of silybin A (0,01%) in medium of suspension culture was found after 24 hours of elicitor treatment in concentration of c1 = 2,33 x 10-3 mol/l.
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Corporate Income Tax in Accounting
Šámalová, Marie ; Müllerová, Libuše (advisor) ; Zelenková, Marie (referee)
The aim of this thesis is to describe in detail the issue of accounting treatment for income tax of legal persons in accordance with current accounting and tax legislation. For a comprehensive understanding of the theory display the corporate accounting is appropriate to first deal with the two systems separately and only then proceed to define the operating results, respectively. tax base as one of the bridges connecting the two systems. Transformation operating results for the income tax base is outside the corporate accounting. Information provided by accounting systems are the most important basis for the determination of the tax at the entity. An integral part of this thesis is a detailed description of the accounting operations, which may be an entity in relation to tax corporate income to meet during the accounting, respectively. period. For clarity, the final part includes practical examples of application of the tax accounting theory.
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A comparison of central and peripheral effect of galantamine.
Galusová, Petra ; Vopršalová, Marie (referee) ; Herink, Josef (advisor)
Galantamine is a reversible inhibitor of acety1cholinesterase (AchE). It is recommended for the treatment of rnild to moderate Alzheimer's dementia, vascular dementia and dementia with Lewy bodies. Galantamine is characterized as cognitive enhancer with dual mechanism ofaction (it has also shown activity in modulating the nicotinic cholinergic receptor). The aim of this work is to determine the effective inhibitory dose of galantamine on the selected route of administration and comparing its central and peripheral effects of galantamine. Determination to proceed on the basis of the fmdings of AchE activity in the central nervous system (in the frontal cortex, hippocampus, septum, basal ganglia and pituitary gland) and on the contrary peripheral nervous system activity of butyry1cholinesterase has been detected (specifically in the liver and plasma). The results obtained by the Ellman's method showed that galantamine is generally a weak inhibitor and statistically significant effect is found only at the highest dose used. At the highest dose used can be expected adverse effects of peripheral type.
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Imbalance in financing of bed care in the Czech Republic and its impacts
Štancová, Dana ; Durdisová, Jaroslava (advisor) ; Kubelková, Karina (referee)
This master thesis deals with the question of medical treatment financing focusing mainly on analysis of health insurance companies reimbursement rates and their comparison with costs of aftercare beds. Secondary goal of the thesis is comparison of acute care beds costs and aftercare beds costs. The opening chapter discusses relationships between economics and health service and describes structure of health care funding compared to insurace companies financing strategies. Payment mechanism mostly used in the aftercare field is the pay-per-day system, this mechanism is therefore later explained in detail including particular effects on after care beds financing. The second part of the thesis compares insurance reimbursement rates with real costs of one particular facility. The analysis proves that health insurance companies reimbursement rates are wrongly set and reimbursement legislation is being violated.
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Effect of administration of drugs from the group of statins on the atherogenic process / / in an experimental model of atherosclerosis.
Koutníková, Jitka ; Nachtigal, Petr (advisor) ; Trejtnar, František (referee)
The effects statins administration on atherogenic process in the experimental model of atherosclerosis. Mgr. Jitka Koutníková The aim of this rigorous work was to detect and quantify the changes of endothelial expression of VCAM - 1 (the marker of endothelial dysfunction) and endoglin (the marker of angiogenesis) in vessel wall of apoE deficient mice. Endoglin is a part of the receptor complex of the transformation growing factor beta (TGF - beta). Statins are the most considerable substances for the treatment of hyperlipidemia and blood - vessel complications (atherosclerosis). Statins decrease levels of LDL cholesterol and also of triglycerides. Moreover, pleiotropic effects take important part in statin's benefit: the decrease of activity of inflammatory and prothrombotic processes. We used apoE deficient mice as the model of atherosclerosis, fed by standard laboratory diet. We analyzed the levels of total cholesterol in blood and we observed expression of VCAM - 1 and endoglin in aortic sinus and part of aortic arch in all mice. Total cholesterol concentrations were assessed enzymatically by conventional diagnostic methods and spectrophotometric analyses. To display VCAM-1 and endoglin expression in aortic sinus and part of aortic arch we used imunohistochemical methods and for quantification of VCAM-1...
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Assessment of Biological Activity of Colorectal Carcinoma using RT REAL-TIME PCR
Pešta, Martin ; Topolčan, Ondřej (advisor) ; Pešek, Miloš (referee) ; Pecen, Ladislav (referee)
The author in his doctoral thesis, proposed primers and conditions optimized design Quantitative PCR for determining the gene expression of MMP-7, TIMP-1, MMP-2 and TIMP- second Revealed the presence of gene expression of GAPDH TIMP-1 and TIMP-2 in cancer line HT-29 SW480 and SW620. Noted the high level expression of MMP-7 in line HT-29th Expression of the 2-3 orders of magnitude higher than the lines SW480 and SW620. MRNA expression of MMP-2 in line HT-29 noticed. MRNA expression of MMP-2 by the detected lines SW480 and SW620. He found that determining absolute and relative expression in tumor lines is ekvivalentní. significantly higher mRNA expression of MMP-7, TIMP-1, MMP-2 and TIMP-2 in tumor tissue compared to normal tissue. It can be used in therapy. Noticed the presence of correlation of gene expression MMP-7, TIMP-1, MMP-2 and TIMP-2 with survival and DFI. He showed that a higher stage cancer correlated with a higher median momery MMP-2/TIMP-2. 7, Noticed the difference in gene expression between TS, TP and DPD in control and tumor tissue. Scored marginally significant increase in TS expression in colon tumors compared to tumors rectosigmoid and rectum. This finding can be used in treatment decisions. scored marginally significant correlation between the expression of TS and DPD. This finding can be...
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The study of selected sewer network in an urbanized catchment
Korytář, Ivo ; Habr,, Vladimír (referee) ; Raclavský, Jaroslav (advisor)
This paper deals the case study of the chosen sewer system in the village Starý Mateřov. The study solves passportization of sewer, what is the base for creating technical condition mainly for three selected sections. According to the results from the technical conditions of the three selected sections were suggested suitable options of redevelopment. The options are compared in financial way. The last chapter suggests two possibilities how to deal with the sewage. The first option consists of the transferring waste water to the waste water treatment plant in Pardubice - Semtín. The second option suggests building a new waste water treatment plant in Starý Mateřov. The options are compared in financial way.
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Laboratory diagnostics of micrometastases in breast cancer patients
Mikulová, Veronika ; Zima, Tomáš (advisor) ; Svoboda, Marek (referee) ; Průša, Richard (referee)
Introduction: The presence of circulating tumor cells (CTC) in the peripheral blood has been associated with worse prognosis and early relapse in breast cancer patients. CTC determination in the peripheral blood has been considered as a liquid biopsy. The aim of this project was to analyze the presence of CTC followed by their molecular characterization with the potential use not only as a new biomarker for real-time monitoring of therapy efficacy but also as a suitable tool for patient's stratification and individualization of treatment for breast cancer. Methods: A total of 54 patients with diagnosed early breast cancer were enrolled into a prospective study. Ten millilitres of peripheral blood were sequentially collected to test for the presence and characterization of CTC during the follow-up of patients. CTC isolation and detection was performed by AdnaTest BreastCancer™ (AdnaGen AG, Germany), which is based on the detection of EpCAM, HER2 and MUC1 specific transcripts in enriched CTC- lysates. cDNA from isolated CTC has been further used for newly optimized qPCR assays for breast tumor and therapy resistance associated genes: TOP1, TOP2A, CSTD, ST6GAL, KRT19 and reference gene actin. qPCR results have been analyzed by Genex software (MultiD Analysis). Results: 195 blood samples have been...
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