Detection of snRNP assembly intermediates in Cajal bodies by fluorescence resonance energy transfer

Spliceosomal small nuclear ribonucleoprotein particles (snRNPs) are required for pre-mRNA splicing throughout the nucleoplasm, yet snRNPs also concentrate in Cajal bodies (CBs). To address a proposed role of CBs in snRNP assembly, we have used fluorescence resonance energy transfer (FRET) microscopy to investigate the subnuclear distribution of specific snRNP intermediates. Two distinct complexes containing the protein SART3 (p110), required for U4/U6 snRNP assembly, were localized: SART3•U6 snRNP and SART3•U4/U6 snRNP. These complexes segregated to different nuclear compartments, with SART3•U6 snRNPs exclusively in the nucleoplasm and SART3•U4/U6 snRNPs preferentially in CBs. Mutant cells lacking the CB-specific protein coilin and consequently lacking CBs exhibited increased nucleoplasmic levels of SART3•U4/U6 snRNP complexes. Reconstitution of CBs in these cells by expression of exogenous coilin restored accumulation of SART3•U4/U6 snRNP in CBs. Thus, while some U4/U6 snRNP assembly can occur in the nucleoplasm, these data provide evidence that SART3•U6 snRNPs form in the nucleoplasm and translocate to CBs where U4/U6 snRNP assembly occurs

The elucidation of the causes of neurogenetic diseases by the MPS data analysis using advanced algorithms

7 Souhrn V rámci disertační práce "Objasňování příčin neurogenetických onemocnění ana- lýzou dat z MPS pomocí moderních algoritmů" jsme zpracovávali MPS data sek- venovaná pomocí panelu genů, celoexomového (WES) a celogenomového (WGS) sekvenování. Sekvenování panelem genů Při sekvenování pomocí panelu genů jsme využívali na našem pracovišti navržený panel genů, které jsou asociovány s onemocněním. Obecnou podmínkou pro zařazení genu do panelu jsou minimálně dvě nezávislé publikace asociující gen s onemocněním a nebo alespoň jedna publikace popisu- jící kauzální varianty v genu ve dvou nebo více nepříbuzných pacientech. Těmto kritériím v případě panelu pro epileptickou encefalopatii (EE) vyhovovalo t.č. 112 genů. Sekvenování pomocí panelu genů bylo provedeno u 257 pacientů s epileptickou encefalopatií. Patogenní či pravděpodobně patogenní variantu jsme nalezli u 28 % případů (72 z 257 pacientů). U patogenních a pravděpodobně patogenních 76 variant jsme provedli další ana- lýzu variant - rozdělili jsme varianty dle genů do skupin dle dědičnosti a dle původu varianty na de novo, zděděné a s neznámým původem. Ze 112 genů v panelu jsme nalezli patogenní nebo pravděpodobně patogenní variantu ve 33 genech, z nich se nejčastěji jednalo o geny s autozomálně dominantní dědičností (50 variant ve 22 genech). Dle...8 Summary The thesis "The elucidation of the causes of neurogenetic diseases by the MPS data analysis using advanced algorithms" is focused on processing the massively parallel sequencing (MPS) data from a gene panel, whole-exome sequencing (WES) and whole-genome sequencing (WGS). The aim of the study was to develop a suitable pipeline to evaluate at least 250 MPS gene panel data, 150 WES data and 20 WGS data in order to improve molecular genetic testing of rare neurogenetic disorders. Associated data management and database implementation is also described. Targeted gene panel sequencing A custom-designed gene panel consisting of ge- nes previously associated with the disease was used. In the Epileptic Encephalopathy (EE) panel, two prerequisites need to be met for inclusion into the panel: the gene has to have been published in at least two independent publications OR at least in one publication but in multiple independent families. In the case of the EE panel, 112 genes were included. The targeted gene panel sequencing was then performed on 257 patients with EE. Pathogenic or likely pathogenic (according to ACMG criteria) variants have been found in 28% of patients (72 out of 257). Further analysis of the pathogenic or likely pathogenic variants was performed (76 in total); the variants were grouped by...Department of Paediatric NeurologyKlinika dětské neurologie2. lékařská fakultaSecond Faculty of Medicin

Appeals in criminal proceedings

APPELS IN CRIMINAL PROCEEDINGS ABSTRACT The goal of this Diploma thesis is to analyse the legal regulation of appeals in criminal proceedings and the issue of this regular remedial measure in general. Author's goal is to present readers with the historical genesis of the institution of appeal since early feudalism until the adoption of current Criminal Procedure Code, considering the most important amendments of this law, and to put the intrastate legal regulation into the context of international law and embedment of the right of appeal in criminal proceedings in international treatises. The author also aims to explain appeal as regular remedial measure together with other remedial measures in Criminal Procedure Code and to introduce basic and specific principles, that impact the legal regulation. In the main chapter the recent regulation of appeals in Criminal Procedure Code is critically analysed. The author further deals with the admissibility and effects of the appeal, by all parties that are legitimate to appeal, the time limit, place and means of filling it, while listing practical examples of abnegation of the right to appeal to a higher court and withdrawal of the appeal, content requirements and possible problems in practice, the operation of the court of first instance after appealing, and also...ODVOLÁNÍ V TRESTNÍM ŘÍZENÍ Cílem předkládané diplomové práce je zanalyzovat právní úpravu odvolaní trestním řízení a obecně problematiku tohoto řádného opravného prostředku. Autor si dává za úkol podat čtenáři informaci o historické genezi institutu odvolání od ranného až do přijetí současného trestního řádu s přihlédnutím k nejvýznamnějším novelám tohoto zákona. Zasadit vnitrostátní úpravu do kontextu mezinárodního práva zakotvení práva na odvolání v trestním řízení v mezinárodních smlouvách. Vyložit odvolání jakožto řádného opravného prostředku z teoretického tedy opravný prostředek charakterizovat, porovnat s ostatními opravnými prostředky trestním řádu a přiblížit základní a specifické principy, které na právní úpravu odvolání dopadají. Ve stěžejní kapitole je kriticky rozebrána recentní úprava odvolání v trestním řádu. Autor se postupně zabývá přípustností a účinky odvolání, všemi osobami, jež jsou oprávněné k podání odvolání, lhůtou, místem a způsobem jeho podání uvedením praktických příkladů, vzdáním se práva na odvolání a zpětvzetím odvolání, obsahovými náležitostmi odvolání a možnými úskalími v praxi, činností soudu prvního stupně po podání odvolání a v neposlední řadě řízením u odvolacího soudu a zákonnými mantinely jeho rozhodování, a to vše s kritickými názory autora, odborníků trestněprávní...Department of Criminal LawKatedra trestního právaFaculty of LawPrávnická fakult

Nano-mechanical properties and morphology of irradiated glass fiber filled polypropylene

Cross-linking is a process in which polymer chains are associated through chemical bonds. Radiation, which penetrated through specimens and reacted with the cross-linking agent, gradually formed cross-linking (3D net), first in the surface layer and then in the total volume, which resulted in considerable changes in specimen behavior. This paper describes the effect of electron beam irradiation on the surface properties (nano-indentation test) of glass fiber filled polypropylene (30%). These nano-mechanical properties were measured by the DSI (Depth Sensing Indentation) method on samples which were non-irradiated and irradiated by low (33, 66 and 99 kGy) and high (132 and 165 kGy) doses of the β – radiation. Nano-indentation test was performed at (400mN) indentation loads. The purpose of the article is to consider to what extent the irradiation process influences the resulting nano-mechanical properties measured by the DSI method. The polypropylene tested showed significant changes of indentation hardness and modulus. The best results were achieved by irradiation at doses of 99 and 132 kGy (increase about 68%) by which the highest nano-mechanical properties of filled polypropylene were achieved. These changes were examined and confirmed by X-ray diffraction and Gel content. © 2017, World Scientific and Engineering Academy and Society. All rights reserved.CZ.1.05/2.1.00/03.0089, ERDF, European Regional Development Fun

The Influence of runner system on production of injection molds

This experimental study describes the influence of runner system on rheological properties during the injection molding process. Economic effects on the amount of production are discussed as well. Autodesk Moldflow Synergy 2016 (Moldflow) was used for the study of the injection process. Three suggestions of the runner system, cold runner system, hot runner system and the combination of cold hot runner system have been promoted. These three variants underwent the rheological and economic analysis. As a result, recommendations for the application of the runner system for the required amount of production have been suggested.Ministry of Education, Youth and Sports of the Czech Republic within the National Sustainability Programme [LO1303 (MSMT-7778/2014)]; European Regional Development Fund under the project CEBIA-Tech [CZ.1.05/2.1.00/03.0089]; TBU in Zlin [IGA/FT/2016/010

Nano-hardness of electron beam irradiated polyamide 11

Cross-linking is a process in which polymer chains are associated through chemical bonds. Radiation, which penetrated through specimens and reacted with the cross-linking agent, gradually formed cross-linking (3D net), first in the surface layer and then in the total volume, which resulted in considerable changes in specimen behaviour. In this study there was found that ionizing beta radiation increased the nano-mechanical properties of polyamide 11 (PA11). The measurement results indicated that ionizing beta radiation (66kGy, 132kGy and 198kGy) was a very effective tool for improvement of indentation hardness, indentation modulus, indentation creep and deformation works of studied polymers. The best results were achieved by irradiation at doses of 132 kGy (increase about 40%) by which the highest nano-mechanical properties of PA11 were achieved.TBU in Zlin [IGA/FT/2016/010]; Ministry of Education, Youth and Sports of the Czech Republic within the National Sustainability Programme [L01303, (MSMT-7778/2014)]; European Regional Development Fund under the project CEBIATech [CZ.1.05/2.1.00/03.0089

Micro-hardness of PBT influenced by beta radiation

Cross-linking is a process in which polymer chains are associated through chemical bonds. Radiation, which penetrated through specimens and reacted with the cross-linking agent, gradually formed cross-linking (3D net), first in the surface layer and then in the total volume, which resulted in considerable changes in specimen behaviour. This research paper deals with the utilization of electron beam irradiated PBT on the micro-indentation test. Radiation doses of 66, 132 and 198 kGy were used for unfilled PBT with the 5% crosslinking agent (triallyl isocyanurate). Individual radiation doses caused structural and micro-mechanical changes which have a significant effect on the final properties of the PBT tested. The highest values of micro-mechanical properties were reached radiation dose of 132 kGy, when the micro-mechanical values increased by about 30%.TBU in Zlin [IGA/FT/2016/010]; Ministry of Education, Youth and Sports of the Czech Republic within the National Sustainability Programme [LO1303 (MSMT-7778/2014)]; European Regional Development Fund under the project CEBIA-Tech [CZ.1.05/2.1.00/03.0089

Polymer fluidity influenced by type and amount of filler

Delivery of polymer melts into the mold cavity is the most important stage of the injection molding process. This paper shows the influence of cavity surface roughness and technological parameters on the flow length of polymers into mold cavity. Application of the measurement results may have significant influence on the production of shaping parts of the injection molds especially in changing the so far used processes and substituting them by less costly production processes which might increase the competitiveness of the tool producers and shorten the time between product plan and its implementation. This research focused into the influence of technological parameters on filling of the injection mold cavity and the flow length respectively.Ministry of Education, Youth and Sports of the Czech Republic within the National Sustainability Programme [LO1303 (MSMT-7778/2014)]; European Regional Development Fund under the project CEBIA-Tech [CZ.1.05/2.1.00/03.0089]; TBU in Zlin [IGA/FT/2016/010

Influence of length of glass fibers in recycled polypropylene on tensile properties

The purpose of this article is to show the influence of grinded glass fibers in polymer material. Tested samples were processed by injection molding. Waste from this production was subsequently ground and a mixture was obtained with different size of crushed material. These particles contains glass fibers of similar length as the specific size of particle. Thus very small particles contain very short fibers. Investigation was carried out by testing tensile properties on material polypropylene with 30% glass fibers. Testing was conducted at ambient temperatures. The effect of reprocessing of material is mainly represented by fiber shortening. As a result, considerable reduction of tensile properties can be observed. The reduction is most evident in the modules of elasticity and ultimate tensile strength.TBU in Zlin [IGA/FT/2016/010]; Ministry of Education, Youth and Sports of the Czech Republic within the National Sustainability Programme [LO1303 (MSMT-7778/2014)]; European Regional Development Fund under the project CEBIA-Tech [CZ.1.05/2.1.00/03.0089

Micro-hardness of surface layer of irradiated Polybutene Terephthalate (PBT)

Using high doses of beta radiation for polybutylene terephthalate (PBT) and its influence on the changes of micromechanical properties of surface layer has not been studied in detail so far. The specimens of PBT were made by injection moulding technology and irradiated by high doses of beta radiation (0, 132, 165 and 198 kGy). The changes in the microstructure and micromechanical properties of surface layer were evaluated using WAXS and instrumented microhardness test. The results of the measurements showed considerable increase in micromechanical properties (indentation hardness, indentation elastic modulus) when high doses of beta radiation are used.Ministry of Education, Youth and Sports of the Czech Republic within the National Sustainability Programme [LO1303 (MSMT-7778/2014)]; European Regional Development Fund under the project CEBIA-Tech [CZ.1.05/2.1.00/03.0089]; internal grant of TBU in Zlin [IGA/FT/2016/010]; Ministry of Education, Youth and Sports of the Czech Republi