Eukaryotic initiation factor 4E (eIF4E) is a key part of initiation and regulation of translation in human cells. Three members of human eIF4E proteins have been characterized: eIF4E1, eIF4E2 and eIF4E3. Cellular stress causes translation initiation inhibition followed by disassembly of the polysomes, those processes are accompanied by the assembly of cytoplasmic RNA granules, called stress granules (SG). Stress granules are dynamic structures whose composition may vary depending on the cell type and the stress stimulus. In this study, human cells were subjected to the following stress conditions: high temperature (HS), sodium arsenite (AS) or hypoxia. Using fluorescence microscopy, pairs of human translational initiation factors from the 4E protein family were visualized and their localization to SG was assessed with one GFP- 4E incorporated in the stable cell line and the other one detected endogenously. Here we show eIF4E1 being a part of all the SGs, both in HS and AS conditions. Next, the eIF4E1 and eIF4E3 proteins together form more SGs than proteins eIF4E1, respectively eIF4E3, with eIF4E2. And last, that the presence of the particular 4E protein has no effect on the composition of SGs. Furthermore, selected groups of proteins were assessed for their potential to localize to the SGs under HS...

5 Abstract Five substances from the group of so-called personal care products, known for their low degradability and regular environmental detection, were tested for toxicity using two fish tissue lines (RTgill-W1 a RTG-2) isolated from rainbow trout (Oncorhynchus miykiss). The tested substances were hexadecylpyridinium chloride (HDP), chlorhexidine (CHX), octenidine (OCT), thymol (THM) and triclosan (TCS). A cell viability assay was performed with each of these compounds using Alamar Blue ™ (AB), 5-carboxyfluorescein diacetate acetoxymethyl ester (CFDA-AM) and neutral red (NR) protocols. The results were used to construct dose-response curves along with an EC50 value for each of these substances. The EC50 values ranged from 0,51 (HDP) to 33,75 µg.ml-1 (THM) for RTgill-W1 and from 0,31 (HDP) to 33,37 µg.ml-1 (THM) for RTG-2. The theoretical LC50 estimation was calculated according to Tanneberger et al. (2013). For all substances, cytochrome P450 1A activity was monitored using 7-ethoxyresorufin-o-deethylase (EROD), four out of five tested chemicals were statistically positive for EROD, the highest EROD response was observed for the most toxic compound - HDP. Only TCS did not show statistically significant cytochrome P450 1A activity. In addition, oxidative stress was measured with the fluorescent dye...

Trichomonads (Parabasalia) are anaerobic microeukaryotes classified in the supergroup Excavata. Inclusion of parabasalids within Excavata is exclusively based on the molecular- phylogenetic evidence. Over 400 species of parabasalids have been described so far, and the vast majority of them are endobiotic. In contrast, only few species of free-living parabasalids forming four independent lineages have been described (Pseudotrichomonas keilini, Ditrichomonas honigbergii, Monotrichomonas carabina, Honigbergiella sp., Tetratrichomonas undula, and Lacusteria cypriaca). Lacusteria cypriaca is a new species and genus described in our recent paper. In this paper we published the first two sequences of SSU rDNA from Pseudotrichomonas keilini as well. All of these lineages are likely secondarily free-living, and they developed from endobiotic ancestors. In addition to the already published Lacusteria cypriaca and Pseudotrichomonas keilini strains, we have recently obtained seven another isolates of free-living trichomonads (LAGOS2D, E2NT, CK, LAGOS2M, GR8, GOU23 LIVADIAN, and VAV1A1); from all of these isolates we sequenced SSU rDNA and performed phylogenetic analyses. These isolates split into four independent evolutionary lineages, which indicate that free-living parabasalids are more diversed and...

Multiple myeloma is an incurable fatal neoplasm of plasma cells affecting mainly elderly people. There are many research laboratories in the world where multiple myeloma is studied. Permanent cell lines are indispensable tools for both basic and applied research. However, the establishment of new cell lines is difficult with poor success. We established 96 primary cultures of bone marrow samples from myeloma patients. Only one culture succeeded in permanet myeloma cell line. The novel plasmacytic cell line ÚHKT-893 was established from bone marrow sample of 57 years old female relapsed with multiple myeloma of IgGκ isotype. The cell line is however dependent on the continuous presence of interleukin-6 in culture media. ÚHKT-893 cells grow continuously already more than 1 year. The growth of cells was regularly monitored according to growth curves and by measurement of cell viability. Surface antigenic profile was repeatedly determined by flow cytometry. The simultaneous expression of both CD138 and CD38 surface molecules confirmed the plasma cell origin of cells. The establishment of the ÚHKT-893 myeloma cell line will extend the panel of existing myeloma cell lines as a new ex vivo model for the study of the etiology and pathogenesis of multiple myeloma. It will also provide the source of new...

Genetic information must be protected, maintained and copied from cell to daughter cells, from generation to generation. In plants, most of the cells contain complete genetic information, and many of these cells can regenerate to a whole new plant. Such a feature leads to the need for precise control of which genes will be active and which not because in growth and differentiation, only the activity of specific genes for the individual cells, tissues, organs are required. One of the mechanisms controlling the gene activity is RNA interference (RNAi), which down- regulates or blocks the expression of specific genes at the transcriptional or post-transcriptional level. The crucial part of the RNAi is guiding the RNAi machinery to the target. It is mediated via sequence complementarity of the target with a small RNA (sRNA), which is diced from a double- stranded RNA (dsRNA) precursor. The molecular mechanism of dsRNA and sRNA formation and also the target origin predestinates the subsequent silencing pathway. In transcriptional gene silencing (TGS), the gene expression is regulated through chromatin epigenetic modifications. One of the epigenetic marks is cytosine methylation, which is established mainly by RNA-directed DNA-methylation (RdDM) pathway. Although the protein machinery was relatively...

This work documents the author's interest in drawing and is based on the mental background of his artistic creation. The work is guided by the interest in the physical conditions of the drawing itself (materiality of the drawing), but also in the study of drawing as a fluid material of artistic creation, entering into the relationship with the image (drawing as a material). At the same time it deals with the process of creation itself and the phenomenon of repetition. The primary goal of the whole work is not the deliberate laying of pre-formulated questions and the subsequent search for their answers, the presentation of some finished truths, or the mere illustration of a certain issue, but rather the process of finding questions that emerge during the process of creation itself.

Regulatory T cells (Treg) play a key role in maintaining the immune tolerance. They suppress development of autoimmune diseases and contribute to maintaining the homeostasis of the immune system. Expansion and excessive ability of regulatory T cells to suppress the immune response is increasingly observed also at many types of cancer. Due to the active inhibition of the antitumor immune response Treg contribute to tumor progression. Specific phenotype based detection and analysis of Treg functional properties may contribute to the successful monitoring of Treg accounts and to the effective cancer immunotherapy itself. Tumor cells express high amounts of so-called tumor antigens, which may play a key role in the antitumor immune response. Expression level of the tumor antigens gives the evidence about relevancy of each antigen in the specific immune response and efficiency of cancer immunotherapy. These data are obviously important to be obtained from the tumor cell lines as well as primary tumor cells. In the first part of the thesis I was focusing on the quantitative analysis of regulatory T cells in tumor tissue and peripheral blood of patients with ovarian cancer. For this purpose I used the newly introduced methyl-sensitive quantitative PCR (MS-qPCR) method and compare the data with the widely...

The existence and widespread distribution of sexual reproduction despite its obvious disadvantages is an evolutionary enigma. According to the theory of frozen plasticity, the evolution of sexual and asexual species is fundamentally different. One of the implications of this theory are different ecological preferences of these species. Sexual species should prefer biotically and abiotically variable environments because of their ability to quickly and reversibly respond to unpredictable changes. On the other hand, asexual species should prosper in biotically and abiotically stable environments because of their ability to perfectly adapt even to extreme environmental factors in the long term. I decided to test these predictions in the metastudy comparing ancient asexual clades (in which we can be sure about their obligate asexuality and evolutionary longevity) with their sexual sister or closely related ecologically comparable clades. The hypothesis of the preference of asexual species to biotically and abiotically homogenous environments was supported by the results of this metastudy. On the other hand, the hypothesis of supposed ability of asexual species to perfectly adapt to a broader range of environmental conditions, tested on the comparison of temperature ranges of activity of previously...

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Die Arbeit stützt sich überwiegend auf die Ergebnisse experimenteller Untersuchungen. Einaxiale Zugversuche an scheibenförmigen Probekörpern bildeten die Basis für die Beschreibung des Tragverhaltens in Form von Spannungs-Dehnungs-Linien. Da die Durchführung von Zugversuchen bekanntlich nicht ganz problemlos ist, wird der Versuchsaufbau besonders sorgfältig dargestellt. Mängel und deren Auswirkung auf die Versuchsergebnisse werden ausführlich diskutiert. Für die Beurteilung der Verbundeigenschaften der Bewehrung ist der Umfang der Kontaktfläche mit der Matrix ein wichtiger Parameter. Es wurde ein Verfahren für die Ermittlung dieser und anderer geometrischen Eigenschaften der Bewehrung entwickelt. Elektronische Aufnahmen von klassischen Dünnschliffen werden dazu mit einem digitalen Bildauswerteprogramm analysiert. Die Berücksichtigung einer Vielzahl von Einflüssen ermöglicht erstmals die reproduzierbare Bestimmung des Umfanges der Kontaktfläche zwischen den Fasern und der Matrix sowie die Querschnittsfläche der Faserbündel in der Matrix ohne subjektive Einflüsse. Das Tragverhalten von unidirektionalen und textilen Bewehrungen aus AR-Glas Filamentgarnen wird mit experimentellen Methoden untersucht. Die Basis bilden in einaxialen Zugversuchen an Dehnkörpern aufgenommene Spannungs-Dehnungs-Linien. Schon bei einer Bewehrung aus unidirektionalen Multifilamentgarnen treten Phänomene auf, die von Bewehrungen aus kompakten, homogenen Querschnitten (Stäben oder Drähten) nicht bekannt sind. Die Multifilamentgarne können selbst schon als Verbundwerkstoff bezeichnet werden, dessen Eigenschaften in einem großen Wertebereich variieren. Mit der textilen Verarbeitung von Multifilamentgarnen werden die (Verbund¬¬-)Eigenschaften entscheidend verändert. Ganz wesentlich ist das Hinzufügen von weiteren Faserscharen und den für die Herstellung der Textilien notwendigen Nähgarnen. Durch die Verarbeitung ändern sich auch Form und Packungsdichte der Multifilamentgarne. Das Zusammenspiel all dieser Einflüsse und die Variationsmöglichkeiten bei der Kombination aller Parameter führen zu einem teilweise deutlich verändertem Tragverhalten des textilbewehrten Betons gegenüber unidirektional bewehrtem Beton. Auf der Basis einfach handhabbarer Modelle wird versucht, Charakteristika des Tragverhaltens zu erklären. Beim textilbewehrten Beton ist jedoch oft kein eindeutiger Zusammenhang erkennbar, weil sich mehrere Einflüsse überlagern. Trotzdem ist es gelungen, zahlreiche, für den textilbewehrten Beton typische Besonderheiten aufzuzeigen und deren Ursachen zu klären. Den Abschluss bilden Überlegungen zur Optimierung der textilen Bewehrungen, die sich aus den beobachteten Phänomenen und deren Ursachen ableiten. / This thesis is mainly based on results of experimental methods. Uniaxial tension tests on strain specimens provide a basis for a description of the load bearing behaviour in the form of stress-strain-curves. It is well known, that the accomplishment of tension tests causes several problems. Therefore the test set-up is described in detail. Deficiencies and their impact on test results are discussed extensively. For the assessment of bond properties the perimeter of the contact area between fibres and matrix is a crucial parameter. A new procedure has been developed for estimating this and other geometrical properties of the reinforcement. Digital images of traditional thin section petrography are analysed with digital image analysis software. The consideration of a multitude of influences enables a reproducible determination of the perimeter of the contact area between fibres and matrix as well as the cross sectional area of the fibre bundles for the first time without subjective influences. The load bearing behaviour of unidirectional fabric reinforcement made from AR-glass filament yarns is examined with experimental methods. Stress-strain-curves from uniaxial tension tests on strain specimen provide a basis. Reinforcements of unidirectional multi filament yarns show already phenomenon?s that are not known from reinforcements with compact homogeneous cross sections (bars or wires). Multi filament yarns could be indicated as an composite material itself whose properties vary in a wide range. During fabric production with textile technologies the (bond-)properties are subject to crucial changes. The adding of extra layers of yarns and needle thread required for connecting these layers are of vital importance. Textile processing changes shape and packing density of the multi filament yarns. The interaction of all these influences and the possible range of combining all parameters lead to pronounced changes in the mechanical properties of textile reinforced concrete also compared to unidirectional fibre reinforcement. Using simple to handle models this work attempts to explain characteristics of the mechanical behaviour. However, for textile reinforced concrete there is often not a clear connection visible. Although numerous typical characteristics of textile reinforced concrete and their causes has been identified. Finally some considerations for optimising fabric reinforcements are given, which has been derived from observed phenomenons and their causes.