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Flowering of Day-Neutral Tobacco Nicotiana Tabacum L. as Influended by Transformation with CDC25
Vojvodová, Petra
Plants are sessile organisms and so they cannot escape changes in environmental conditions. In order to optimize seed production and to ensure that flowering occurs during the appropriate season, plants have evolved a complex of regulatory pathways to control when the floral transition takes place. One group of regulatory pathways involves enviromental factors like certain photoperiod or vernalization. The second group of regulatory pathways involves the state of development and is not sensitive to environmental cues, this pathway is called autonomous pathway and plants in this group do not require a particular photoperiod or vernalization, they are called day-neutral plants. The transition of these plants from vegetative to reproductive phase depends on combination of reaching certain minimal number of nodes, inhibition effect of roots, floral signals generated by leaves and activity of the shoot apical meristem (SAM). Among early events of the transition of SAM to reproductive phase belongs an increase in the mitotic activities of the cells in SAM. Precise mechanisms of the action of the signals connected with transition of SAM to flowering have not been elucidated yet. The plants of day-neutral tobacco (Nicotiana tabacum L.) cv. Samsun were transformed with fission yeast mitotic activator coded...
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Optimization of the Synthesis of 3-(4-Bromophenyl)-5-hydroxymethyl-5,6-dihydro- pyran-2-one
Krenk, Ondřej ; Pour, Milan (advisor) ; Klimešová, Věra (referee)
Treatment of cancer still requires searching of new antineoplastics. Studies of biological activity of natural produkts show, that wide spektrum of biological aktivity have α,β- unsaturated-δ-laktons as potential substance with cytostatik aktivity. Goal of this work is optimalization of synthesis of 3-(4-bromfenyl)-5-hydroxymethyl-5,6-dihydro-2H-pyran-2-one as analogue of biologically active substance with potential cytostatic effect against cell line of colorectal carcinoma. Results obtained from this work can be employed in the development of simplier and more economical synthesis of potential biologically active analogues of α,β- unsaturated-δ-lactones.
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Study of transport mechanisms of xenobiotics
Richtrová, Monika ; Trejtnar, František (advisor) ; Melicharová, Ludmila (referee)
Aim: Graduation theses was aimed on the use of the isolated rat renal cells to study of transport mechanisms of the selected receptor - specific peptides from the group of somatostatin analogues labeled with convenient radionuclides - 111In-DOTA-octreotate, 125IDOTA- octreotate. Experiment with the selected model substances - sucrose and α-methyl glucoside as markers of active and passive transport was executed. Accumulation process of α-methyl glucoside was tested in renal and pancreatic cells. Further, accumulation of the radiopeptides was researched in presence of the potential inhibitors, which could reduce to retention these radiopeptides and thus also clinically undesirable renal radiotoxic insult. Methods: Isolated rat renal cells were prepared by collagenase technique. Pancreatic rat exocrine cells were prepared from cell line tumor cells. Viability of cells was tested with trypane blue. The accumulation rate of model substances was compared with the accumulation radiopeptides rate: 111In-DOTA-octreotate, 125I-DOTA-octreotate. Transport character was also monitored for low temperatures. Megalin/cubilin membran transport system was researched via their ligands (e.g. albumin, gentamicin), which could inhibit accumulation of the radiopeptides by competition in transport. Results: Cells...
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Plasmide DNA isolation from bacteria and transfection to HEK293 cell line
Skala, Kateřina ; Fohlerová, Zdenka (referee) ; Svoboda, Ondřej (advisor)
DNA isolation is one of the basic methods in molecular biology. There are several methods of DNA amplification and isolation. In this paper phenol-chloroform extraction of three plasmid types - Channelrhodopsin-2, ASAP1 and Kir 2.1 is used. Six plasmids were isolated in total. These plasmids are then validated using gel electrophoresis. Successfully isolated plasmids are then transfected to HEK293 cells and images taken on confocal microscope 24, 48 and 72 hours after transfection.
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Investigation of chlamydia urogenital tract
BENDOVÁ, Radka
The subject of this bachelor paper is chlamydiae examination in urogenital tract via the method of cultivation. The aim of this paper is to test the cultivation method of diagnosis Chlamydia trachomatis on a tissue of culture, to evaluate the outcome according to individual criteria in a given period and to compare the outcome with the literary data. The virology laboratory in hospital in České Budějovice, a.s. Is equiped for the culture tissue preparation and for this method they use uncommon line of cells BGM, which is not commonly used in the world. In this department this line has proven and so they use it since 1985, when they received it. This paper informs about the present situation of chlamydiae, their clasification according to phylogenetic and genome bacterial analysis, about their occurence, growth cycle and multiplication. The paper also focuses on chlamydiae infections divided into ocular and urogenital infection and their treatment. The overview of present issues is finished by the methods of evidence, direct and indirect evidence. In the direct evidence there are described the issue cultures their preparation and cultivation. This method was verified in the practical part of my bachelor paper. For this bachelor paper I was provided with samples from urogenital tract which were evaluated in three categories: positive, negative and imposible to evaluate (samples were overgrown with bacteria or yeast cells and it was imposible to evaluate them). They were mostly smears from urethra and cervix (at women) or from urethra only (at men). The number of all samples was 159 and they were examined during the period from 18th October to 5th December 2011. Chlamydiae grew on issue cultures for 48 hours and then they were died with modified colouring Macchiavella. 127 examples (79.9%) coloured by this technique were evaluated from the total number. Ambiguous samples were coloured by the monoclonal antidote against Chlamydia trachomatis. 32 samples (20.1%) were coloured using this technique from the total number.I also evaluated the samples according to age. Unusually high was positivity (42.2%) at women at the age of 26-46. Which includes the highest number of samples (54.2%) of women from the total number. It mostly concerned pacients with gynaecological difficulties who were never examined and treated or they had a chronical infection which can be treated with difficulties and inclinates to relapse. At men the highest number of samples (56.1%) from the total number was at the age of 26-46 but only 13% were positive. The difference is given by the size of samples and also at men the occurence of chronical infection is less common.
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Endoplasmic reticulum stress
Červenka, Jakub ; Schierová, Michaela (advisor) ; Horníková, Lenka (referee)
The accumulation of unfolded or misfolded proteins in endoplasmic reticulum (ER) leads to ER stress and the activation of unfolded protein response (UPR). Recent studies show that ER stress or UPR are associated with many diseases such as diabetes, hepatitis type C, prion disease, different kinds of tumors or Alzheimer's, Parkinson's and Huntington's disease and also with physiological processes like cell differentiation. When UPR is activated in yeast Saccharomyces cerevisiae, Ire1 protein oligomerizes, transautophosphorylates and activates itself. After this, Ire1 cleaves HAC1 mRNA to remove an intron. The spliced form of HAC1 mRNA is translated into the Hac1 transcription factor, which induces transcription of genes for chaperones of lumen ER, proteins involved in ERAD, synthesis of lipids etc. The cell uses this to reestablish homeostasis in ER. In mammals, the UPR is more complex and except Ire1 dependent pathway, it comprises Perk and Atf6 pathways, which are missing in yeast. Nevertheless, Perk is activated and regulated by the similar mechanism as Ire1 in S. cerevisiae. In consideration of broad spectrum of methods for genetic manipulation, rapid growth and well annotated genome, the yeast S. cerevisiae is a useful model for study of general mechanisms of UPR in mammals.
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