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Changes in Expression of Membrane Molecules CD200R, CD95, CD95L, and Soluble CD200R Regulating inflammatory Responses in Patients Undergoing Cardiac Surgery
Holmannová, Drahomíra ; Krejsek, Jan (advisor) ; Brát, Radim (referee) ; Turánek, Jaroslav (referee)
Cardiac surgery is known to initiate a complex physiological response with the immune system activation (SIRS), neurohormonal response, metabolic changes, coagulopathies etc. SIRS is triggered by tissue injury, myocardial ischemia, reperfusion, use of anaesthesia, cardioplegia, extracorporeal circuit etc. Excessive immune system activation is associated with progression of SIRS, life-threatening multi-organ dysfunction (MOD), and increased morbidity/mortality in the postoperative period. The immune system response is regulated and terminated by both cellular and humoral regulatory and inhibitory mechanisms including changes in expression of in our study monitored molecules: CD200/CD200R, sCD200R and CD95/CD95L. Methods: The study included the measurement the expression of CD95, CD95L, CD200R, and sCD200R molecules in granulocyte and monocyte populations in blood samples of 30 patients who underwent heart surgery using CPB. Samples collected before surgery, after surgery, and in the postoperative period (1st , 3rd , 7th day) were analysed by flow cytometry and sCD200R by ELISA. Results: We discovered a significant increase in the percentage of granulocytes expressing inhibitory molecule CD200R (from 5% to 17.8%) instantly after surgery. It might be presumed that these cells are less susceptible to...
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Viral vectors based on tobacco mosaic virus for the expression of recombinant proteins in plants
Kroupa, Daniel ; Moravec, Tomáš (advisor) ; Cvrčková, Fatima (referee)
(eng): Over the past few decades, plant viruses and vectors derived from them have become not only an integral part of the work in laboratories, but also began to be used commercially for the production of vaccines, drugs, enzymes and other proteins / peptides. One of the most commonly used viruses for the expression of heterologous proteins is tobacco mosaic virus. In this work, I would like to focus on plant viruses and vectors derived therefrom. Compare it with other expression systems used in plants and describe how their benefits could be further used and combined with modern methods of cloning. Key words: viral vector, expression, Tobacco mosaic virus, plant expression systems, Golden gate cloning
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Characterization of eukaryotic translation initiation factor 3 subunits (eIF3) in A. thaliana male gametophyte
Linhart, Filip ; Hafidh, Said (advisor) ; Retzer, Katarzyna (referee)
From RNA-to-protein, translation initiation and protein synthesis is mediated by trans-acting factors that recognize mRNA features common to almost all eukaryotes. Eukaryotic translation initiation factor 3 complex (eIF3) is a highly conserved protein complex that recognizes 5'-CAP elements of the mRNA to initiate translation. eIF3 consists of nine subunits, three of them having two isoforms: eIF3A, eIF2B1, eIF3B2, eIF3C1, eIF3C2, eIF3D, eIF3E, eIF3F, eIF3G1, eIF3G2, eIF3H and eIF3K. This work deals with functional characterization, expression and subcellular localization of eIF3B1, eIF3B2 and eIF3E in Arabidopsis thaliana male gametophyte and interaction of eIF3E with the Constitutive photomorphogenesis 9 (COP9) complex as a regulatory complex of eIF3E post-translational control. Here we show that depletion of eif3b1 or eif3b2 is not gametophytic lethal and that the two protein might function redundantly, whereas, knockout of eIF3E causes male gametophyte lethality. Interestingly, eif3b1 show post-fertilization defects during embryogenesis, suggesting that its redundancy with eIF3B2 is restricted to the gametophyte. Gene expression studies revealed high expression of eIF3 subunits in actively dividing zones of leaf primordia, root meristem and root elongation zones as well as in the vegetative...
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The effect of selected endocrine disruptors on cytochromes P450 1B1 and 3A1/2
Holecová, Jana ; Bořek Dohalská, Lucie (advisor) ; Cajthaml, Tomáš (referee)
Many exogenous and endogenous compounds are referred to as endocrine disruptors (EDCs), as they interfere with natural synthesis, signaling and metabolism of endogenous hormones. Common exogenous endocrine disruptors are benzo(a)pyrene (BaP) and 17α-ethinylestradiol (EE2). Endogenous endocrine disruptor 17β-estradiol (E2) is frequently present in the environment as well. In this thesis, the effect of the mentioned EDCs and their combinations on gene and protein expression of CYP1B1, 3A1 and 3A2 in rat liver, kidney and lung was determined. Protein expression was studied using Western blot method and specific antibodies; gene expression was assessed by quantitative PCR. Moreover, the effect of tested EDCs and their combinations on BaP metabolism and CYP3A specific activity (measured as testosterone 6β-hydroxylation) were studied in liver microsomal samples. It was confirmed, that BaP significantly increases CYP1B1 expression in rat liver and lung both alone and together with EE2 or E2. Pretreatment of rat with E2 and BaP increases the ability of BaP to induce CYP1B1 expression. On the contrary, EE2, E2 and their combination decrease the CYP1B1gene expression. The rate of BaP metabolites formed in liver microsomal samples increases in rats pretreated with BaP and its combinations. In liver, there was...
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The influence of endocrine disruptors on the expression of cytochrome P450 1A2
Orlovská, Ľubica ; Dračínská, Helena (advisor) ; Ječmen, Tomáš (referee)
The term endocrine disruptor is used for chemical compounds which imitate or antagonize the effects of endogenic hormones, alter hormone synthesis and metabolism or modify levels of hormonal receptors. Synthetic estrogen 17α-ethinylestradiol (EE2) and carcinogenic substance benzo[a]pyrene (BaP) belong to the group of chemicals described as exogenic compounds with endocrine destruction, while estrogenic hormone 17β- estradiol (EST) figures as natural endogenic endocrine disruptor. The bachelor thesis focuses on study of the influence of these endocrine disruptors and their combinations on expression and specific activity of CYP1A2. RNA was isolated from the lungs of rats treated with the endocrine disruptors and from untreated rats. RNA was converted to cDNA by reverse transcription. Relative amount of CYP1A2 in livers, kidneys and lungs was quantified by real-time PCR. The protein expression of CYP1A2 was studied using the Western blot with consecutive immunodetection. Finally, the specific activity of hepatic CYP1A2 was determined by measuring 7-methoxyresorufin O- demethylation. It was confirmed that BaP induces gene expression of CYP1A2 in livers, kidneys and lungs, even in combination with EE2 and EST. However, both estrogens decrease the induction potential of BaP. When given individually,...
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Artefiletics activity in preschool education
ČERNÁ, Dana
In my bachelor thesis, I aim to explore the possibility of utilization of artefiletics activities at preschool education, which should lead to personality development of preschool children. This bachelor thesis is divided into two main parts, namely theoretical and practical. The theoretical part deals with development psychology and characterization of preschool age in relation to the artefiletics. In context with the Framework Educational Program, this part also focuses on the artefiletics as one of options of educational supply at a kindergarten. The practical part describes realization of artefiletics activities with preschool children along with children's personal experiences and their expressions. The conclusion contains summary of findings from my research.
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Supervision with Visual Art Expression
Hanzlová, Andrea ; Havrdová, Zuzana (advisor) ; Slavík, Jan (referee)
Title: Supervision with Visual Art Expression Author: Bc. Andrea Hanzlová ABSTRACT Although art expression has been widely used in both art therapy and supervision in the Czech Republic, its distinctive features, added value and possible drawbacks in supervision have been researched less extensively than in art therapy. Here I explore what art expression brings into supervision, what new perspectives it can offer and what obstacles may dissuade supervisors from employing certain approaches. A qualitative content analysis of semi- structured interviews on the sample of 12 respondents revealed four thematic areas: Definition of creative art process, phases and content; Drawbacks and Benefits of working with art expression; The role of education in used methods and approaches. The results show an unexpectedly wide range of methods that Czech supervisors and supervisees employ when dealing with art expression in supervision, as well as the extent or absence of some approaches. One possible reason could be the lack of legislative anchoring compared to countries such as the UK and USA. Key words: expression, visual creative process, artefact, expressive therapy, art therapy, supervision, supervision of art therapies
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Characteristics of expression vectors for Corynebacterium glutamicum and their use for studies of sigma factors of RNA polymerase
Dvořáková, Pavla ; Pátek, Miroslav (advisor) ; Konopásek, Ivo (referee)
The aim of the thesis was to characterize chosen expression vectors used in biotechnologically important bacterial species, Corynebacterium glutamicum, and to test their use in studies of promoter activity control by sigma factors of RNA polymerase. Different properties of these vectors (level of expression of the cloned gene, leaky expression without inducer, dependence of expression level on inducer concentration and cell population homogeneity) were found by determination of expression level of the model gfpuv gene by fluorescence intensity assay of the produced protein and by gfpuv-expressing C. glutamicum cell population analysis using flow cytometry. The vector pEC-XT99A was chosen for testing the bi-plasmid system for assignment of a sigma factor to the chosen promoter. Although the level of expression provided by pEC-XT99A was not high, the vector showed no leaky expression, expression from the vector was comparable for a wide range of IPTG concentrations and the cell population was homogenous concerning the gene expression. Using pEC-XT99A from which individual stress sig genes were expressed, the σD factor was clearly assigned to the up-to-now unknown Pcg0420 promoter. Another vector for isolation and purification of C. glutamicum proteins was used to express the C. glutamicum sigM gene and to...
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John Dewey's Aesthetics and interactive art
Rubášová, Veronika ; Kaplický, Martin (advisor) ; Dadejík, Ondřej (referee)
This bachelor thesis is especially focused on John Dewey's theory. The first and the main part of the bachelor thesis deals with Dewey's conception of experience, with analysis of this process of experience and with aesthetic level of experience. Significant part of bachelor thesis is dedicated to aesthetic experience with art as well, which is discussed in the context of every day experiences. Continuity between both is highlighted. The second part of the bachelor thesis is an addendum to the interpretation of Dewey's theory. This part of the thesis deals with interactive art. Interactive art is interpreted using Dewey's theory. Interactive art is in this paper understood as artistic type which can emphasize continuity between art and everyday experience so as Dewey does.
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The effect of selected endocrine disruptors on the cytochromes P450 1A1 and 2C
Klusoňová, Zuzana ; Bořek Dohalská, Lucie (advisor) ; Linhartová, Lucie (referee)
Many currently produced chemicals reveal specific properties which allow them to be referred to as endocrine disruptors (ED). These substances exhibit an exogenic hormone activity and usually act as antagonists or agonists of endogenic hormones. The exogenic EDs studied in this work were 17α-ethinylestradiol (EE2) and benzoapyrene (BaP). 17β-estradiol (E2), a typical endogenic hormone, was also included to the study. In the presented work, the effect of these EDs and their combinations on the expression and specific activities of cytochromes P450 (CYP) 1A1 and 2C was determined. First, the microsomal fraction (MF) of liver, kidney and lung of rats premedicated with these compounds or without premedication was isolated. CYP expression was assessed by the Western blot analyses in these MF samples. Moreover, CYP1A1 and CYP2C specific activities were evaluated. It was found that premedication of rats with BaP increased CYP1A1 expression in all above mentioned organs. Whereas BaP strongly induced rat CYP1A1, EE2 and E2 were almost without this effect. But, when these disruptors were administered to rats with BaP, they supported its potency to induce CYP1A1. Further, CYP2C11 expression and its specific activity were gently increased by premedication of rat with EE2 and its combination with BaP....
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