Characterizing functional domains of the RNA helicase RHAU involved in subcellular localization and RNA interaction

Posttranscriptional regulation of gene expression is an important and highly regulated process in response to developmental, environmental and metabolic signals. During stress conditions such as heat shock (HS), oxidative stress, ischemia or viral infection, the translation machinery of cells is reprogrammed. The majority of actively translated mRNAs is released from polysomes and driven to specific cytoplasmic foci called stress granules (SGs), where dynamic changes in proteinRNA interaction determine the subsequent fate of mRNAs. In the presented thesis, I show that the DEAH-box RNA helicase RHAU is a novel SG-associated protein and that its N-terminus is necessary and sufficient for localization of RHAU in SGs. While RHAU protein was originally identified as an ARE-associated protein involved in uPA mRNA decay, it was not clear whether RHAU directly interacts with RNA. Here, I demonstrate that RHAU physically interacts with RNA in vitro and in vivo through the N-terminus. Bioinformatic analysis of the RHAU protein sequence corroborates the experimental data, revealing that the N-terminus of RHAU harbors a unique RNA-binding domain consisting of two abutting motifs: the G-rich region containing one RGG-box and the RHAU specific motif (RSM). It is widely believed that substrate specificity and subcellular localization of RNA helicases is mediated by their less conserved flanked N-/Cterminal domains. As the unique N-terminus of RHAU is essential and sufficient for both subcellular localization and RNA interaction, it most probably determines a functional specificity of RHAU. I further show that ATPase activity is responsible for the apparent instability of RHAU-RNA complex formation and markedly influences the kinetics of RHAU retention in SGs. The striking difference in SG shuttling kinetics between fully active RHAU protein and its ATPase-deficient mutant triggers the hypothesis that its ATPase activity takes part in energy dependent dynamic remodeling of RNPs in SGs. In summary, the results presented in this thesis demonstrate that after rck/p54, DDX3 and eIF4A, RHAU is the fourth RNA helicase detected in SGs and that its association with SGs is dynamic and mediated by a RHAU-specific RNAbinding domain. Additionally, I could show that RHAU is an essential factor for P-body (PB) formation and obtained initial data that RHAU is possibly also involved in the process of translation via its interaction with translation initiation factor eIF3b

Types of bilingualism in relation to the manifestations of personality traits

This bachelor thesis deals with bilingualism and its relation to personality expressions. In the literature review section divided into seven chapters, bilingualism is first described and defined. The next part of the thesis focuses on the description of different types of bilingualism, especially on divisions based on according to the age of the second language was acquired. The following chapters describe the different specificities of bilinguals. The last two chapters then focus on the links between bilingualism, personality and biculturalism, summarizing selected research on these relationships. The final part of the thesis contains a proposal for a research project that aims to compare Czech-Russian bilinguals in their personality expressions with respect to the language used. In particular, it involves comparing a group of early and late bilinguals with each other to find out whether they differ in their personality expressions within the two languages.Tato bakalářská práce se zabývá bilingvismem a jeho vztahem k osobnostním projevům. V literárně přehledové části, která je rozdělena do sedmi kapitol, je nejprve popsán a definován bilingvismus. Další část práce se zaměřuje na popis různých typů bilingvismu, a především pak na dělení podle věku osvojení druhého jazyka. Další kapitoly popisují různá specifika bilingvních osob. Poslední dvě kapitoly se pak věnují propojení bilingvismu s osobností a bikulturalitou, kde jsou shrnuty vybrané výzkumy zabývající se těmito vztahy. Další část práce obsahuje návrh výzkumného projektu, který si klade za cíl porovnat česko-rusky mluvící bilingvní osoby v jejich osobnostních projevech v závislosti na použitém jazyce. Především pak mezi sebou porovnat skupinu raných a pozdních bilingvních osob a zjistit, zda se mezi sebou v osobnostních projevech v těchto dvou jazycích liší.Department of PsychologyKatedra psychologieFilozofická fakultaFaculty of Art

Quality control of food supplements with melatonine using separation methods

Charles University Faculty of Pharmacy in Hradec Králové Department of Analytical Chemistry Candidate: Natálie Chalupníková Supervisor: prof. RNDr. Dalibor Šatínský, Ph.D. Title of the diploma thesis: Quality control of food supplements with melatonine using separation methods In this diploma thesis, an HPLC method was developed and validated for the simultaneous determination of melatonin and pyridoxine. The developed method was applied to the determination in food supplements, Tozax Sleep Trip (mcePharma s. r. o.), Clinical Melatonin B6 syrup (Clinical nutricosmetics s. r. o.), Fast Sleep oral spray with melatonin (EmergoPharm Sp. z o. o. Sp. K.), Melatonin Rapid complex 5 mg ODT (Salutem Pharma s. r. o.), Allegra Melatonin 3 mg (Woykoff a. s.), Clinical Melatonin Forte (Clinical nutricosmetics s. r. o.), Melatonin 2 mg (Allnature s. r. o.), Melatonin new extra 5 mg (Nutricius s. r. o.), Clinical Melatonin B6 Gummies (Clinical nutricosmetics s. r. o.). The method was based on the use of a SUPELCO Ascentis Express C-18 chromatography column (100*4.6 mm, particle size 2.7 µm). The separation was carried out under gradient elution, using a mobile phase of acetonitrile and 0.085% phosphoric acid at pH 2.2, flow rate 1 ml/min and constant temperature 30řC. A detection wavelength of 278 nm was chosen...Univerzita Karlova Farmaceutická fakulta v Hradci Králové Katedra analytické chemie Kandidát: Natálie Chalupníková Vedoucí práce: prof. RNDr. Dalibor Šatínský, Ph.D. Název diplomové práce: Kontrola kvality potravních doplňků na bázi melatoninu pomocí separačních metod V této diplomové práci byla vyvinuta a validována HPLC metoda pro současné stanovení melatoninu a pyridoxinu. Vypracovaná metoda byla aplikována na stanovení v potravních doplňcích, Tozax Sleep Trip (mcePharma s. r. o.), Clinical Melatonin B6 sirup (Clinical nutricosmetics s. r. o.), Fast Sleep ústní sprej s melatoninem (EmergoPharm Sp. z o. o. Sp. K.), Melatonin Rapid komplex 5 mg ODT (Salutem Pharma s. r. o.), Allegra Melatonin 3 mg (Woykoff a. s.), Clinical Melatonin Forte (Clinical nutricosmetics s. r. o.), Melatonin 2 mg (Allnature s. r. o.), Melatonin new extra 5 mg (Nutricius s. r. o.), Clinical Melatonin B6 Gummies (Clinical nutricosmetics s. r. o.). Metoda byla založena na využití chromatografické kolony SUPELCO Ascentis Express C-18 (100*4,6 mm, velikost částic 2,7 µm). Separace probíhala za gradientové eluce s využitím mobilní fáze acetonitril a 0,085% kyselina fosforečná při pH 2,2, průtokové rychlosti 1 ml/min a konstantní teplotě 30řC. Pro detekci melatoninu byla zvolena vlnová délka 278 nm a 290 nm pro pyridoxin....Katedra analytické chemieDepartment of Analytical ChemistryFaculty of Pharmacy in Hradec KrálovéFarmaceutická fakulta v Hradci Králov

Faktory ovlivňující míru otevřenosti ekonomik vybraných zemí

Chalupníková, K. Factors influencing the degree of trade openness of selected coun-tries. Diploma thesis. Brno: Mendel University in Brno, 2019. The main aim of this work is to identify and quantify factors influencing trade open-ness in CEEC countries. For this aim is used regression, correlation and panel data analysis. The dataset consists of 11 CEE countries in the period 2002-2017. A com-parison of territorial and commodity structure of selected countries is also per-formed. The empirical analysis showed that the real exchange rate, unemployment, GDP, price index and EU membership have a statistically significant impact on the degree of openness

Effect of metapopulation structure and environmental factors on population biology of Impatiens noli-tangere infected by powdery mildew Podosphaera balsaminae

Studies performed on natural pathosystems revealed that the spatial structure of the metapopulation plays an important role in the development and spread of disease; phytopathological experimental studies have also revealed a significant effect of climatic factors on the presence of the disease (incidence) and the level of infection (prevalence) in populations. The aim of this study was to explore a natural pathosystem from both sides at the same time and to see which local and spatial environmental factors influence the development of pathosystem. Study focuses on population biology of natural pathosystem Podospaera balsaminae - Impatiens noli-tangere, which has not been investigated yet. In the study area (between the city Sázava (Benešov district) and village Vlkančice) host populations occurred in 78 (64 of them were infected) in 2013 and 82 populations (61 of them were infected) in 2014. Size of host populations had the strongest effect to incidence and prevalence of disease in the pathosystem. In larger populations, the disease occurred more often, developed faster and was higher. The rate of spread of the pathogen territory and the rate of population host connectivity had also strong effect. The abundance of pathogen in the studied area and the level of connectivity of host populations also..

The impact of the negative effect of media on primary school pupils from the point of view of media education

In this diploma thesis, I focus on the problem of negative media influence on pupils of primary school from the viewpoint of media education. This thesis consists of two parts {--} the theoretical and the practical one. The theoretical part deals with general media characteristics, its development and its functions, with the emphases on the latest media and television technology. It also discusses the possible negative media impact and influence on pupils of primary school. This part also focuses on these children{\crq}s leisure time which is considered to be a very important field in terms of media education needs and realizations. At the end of this part, the possible solutions are shown, especially in children`s leisure time activities. The practical part includes a survey performed on certain pupils of primary school. The aim of this survey was to find out the level of the children media education exerted from pupils` parents and teachers. The results present children{\crq}s relationship to various medial areas, the parents` interest in their children`s leisure time activities, the parents` influence on which and how much media their children use and, finally, this survey also presents teacher{\crq}s opinions about this topic in terms of media education