203 research outputs found

    Reading comprehension of czech pupils at the beginning of primary school: comparison betweenanalytic-synthetic and genetic methods

    Get PDF
    In Czech schools two teaching methods of reading are used: the analytic-synthetic (conventional) and genetic (created in the 1990s). They differ in theoretical foundations and in methodology. The aim of this paper is to describe the above mentioned theoretical approaches and present the results of study that followed the differences in the development of initial reading skills between these methods. A total of 452 first grade children (age 6-8) were assessed by a battery of reading tests at the beginning and at the end of the first grade and at the beginning of the second grade. 350 pupils participated all three times. Based on data analysis the developmental dynamics of reading skills in both methods and the main differences in several aspects of reading abilities (e.g. the speed of reading, reading technique, error rate in reading) are described. The main focus is on the reading comprehension development. Results show that pupils instructed using genetic approach scored significantly better on used reading comprehension tests, especially in the first grade. Statistically significant differences occurred between classes independently of each method. Therefore, other factors such as teacher´s role and class composition are discussed

    PROMETHEUS IN THE COMPANY OF TEACHERS, STUDENTS AND OWL (ARCHITECTURE AND FINE ARTS ON THE CAMPUS OF VŠB - TECHNICAL UNIVERSITY OF OSTRAVA)

    Get PDF
    The paper deals with the connection between sculptures and architecture in the second half of the 20th century. The research focuses on artistic works in public space of the VŠB – Technical University campus in Ostrava. In the socialist era, during which the campus was established in Ostrava – Poruba, the funds for artwork were allocated directly in capital budgets for each structure. Presently, the purchase and installation of fine artefacts is left to random decision. An example illustrating hereby the connection between the sculpture and architecture is the history of a relief by the sculptor Vladislav Gajda from the years 1968-1974 located above the VŠB-TUO main building entrance

    Quadruplex DNA: sequence, topology and structure.

    Get PDF
    G-quadruplexes are higher-order DNA and RNA structures formed from G-rich sequences that are built around tetrads of hydrogen-bonded guanine bases. Potential quadruplex sequences have been identified in G-rich eukaryotic telomeres, and more recently in non-telomeric genomic DNA, e.g. in nuclease-hypersensitive promoter regions. The natural role and biological validation of these structures is starting to be explored, and there is particular interest in them as targets for therapeutic intervention. This survey focuses on the folding and structural features on quadruplexes formed from telomeric and non-telomeric DNA sequences, and examines fundamental aspects of topology and the emerging relationships with sequence. Emphasis is placed on information from the high-resolution methods of X-ray crystallography and NMR, and their scope and current limitations are discussed. Such information, together with biological insights, will be important for the discovery of drugs targeting quadruplexes from particular genes

    Label-free nanofluidic scattering microscopy of size and mass of single diffusing molecules and nanoparticles

    Get PDF
    Nanofluidic scattering microscopy enables label-free, quantitative measurements of the molecular weight and hydrodynamic radius of biological molecules and nanoparticles freely diffusing inside a nanofluidic channel. Label-free characterization of single biomolecules aims to complement fluorescence microscopy in situations where labeling compromises data interpretation, is technically challenging or even impossible. However, existing methods require the investigated species to bind to a surface to be visible, thereby leaving a large fraction of analytes undetected. Here, we present nanofluidic scattering microscopy (NSM), which overcomes these limitations by enabling label-free, real-time imaging of single biomolecules diffusing inside a nanofluidic channel. NSM facilitates accurate determination of molecular weight from the measured optical contrast and of the hydrodynamic radius from the measured diffusivity, from which information about the conformational state can be inferred. Furthermore, we demonstrate its applicability to the analysis of a complex biofluid, using conditioned cell culture medium containing extracellular vesicles as an example. We foresee the application of NSM to monitor conformational changes, aggregation and interactions of single biomolecules, and to analyze single-cell secretomes

    In vitro replication and thermodynamic studies of methylation and oxidation modifications of 6-thioguanine

    Get PDF
    The cytotoxic effects of thiopurine drugs are mostly exerted through the formation of thioguanine nucleotide and its subsequent incorporation into DNA. The 6-thioguanine (6-TG) in DNA can be converted to S6-methylthio-2-aminopurine (2-AP-6-SCH3) and 2-aminopurine-6-sulfonic acid (2-AP-6-SO3H) upon reaction with S-adenosyl-L-methionine and irradiation with UVA light, respectively. Here we prepared oligodeoxynucleotides (ODNs) harboring a 6-TG, 2-AP-6-SCH3 or 2-AP-6-SO3H at a defined site and examined, by using LC-MS/MS, the in vitro replication of these substrates with yeast polymerase η and Klenow fragment (KF−). Our results revealed that 2-AP-6-SCH3 could be bypassed by KF−, with significant misincorporation of thymine opposite the lesion. The 2-AP-6-SO3H, however, blocked markedly the nucleotide insertion by KF−. Yeast pol η could bypass all three modified nucleosides; although dCMP was inserted preferentially, we found substantial misincorporation of dTMP and dAMP opposite 2-AP-6-SCH3 and 2-AP-6-SO3H, respectively. Moreover, both KF− and yeast pol η induced a considerable amount of -2 frameshift products from the replication of 2-AP-6-SCH3- and 2-AP-6-SO3H-bearing substrates. Our results also underscored the importance of measuring the relative ionization efficiencies of replication products in the accurate quantification of these products by LC-MS/MS. Moreover, thermodynamic studies revealed that 2-AP-6-SCH3 and 2-AP-6-SO3H could cause more destabilization to duplex DNA than 6-TG. Taken together, the results from this study shed important new light on the biological implications of the two metabolites of 6-TG

    RNAMotifScan: automatic identification of RNA structural motifs using secondary structural alignment

    Get PDF
    Recent studies have shown that RNA structural motifs play essential roles in RNA folding and interaction with other molecules. Computational identification and analysis of RNA structural motifs remains a challenging task. Existing motif identification methods based on 3D structure may not properly compare motifs with high structural variations. Other structural motif identification methods consider only nested canonical base-pairing structures and cannot be used to identify complex RNA structural motifs that often consist of various non-canonical base pairs due to uncommon hydrogen bond interactions. In this article, we present a novel RNA structural alignment method for RNA structural motif identification, RNAMotifScan, which takes into consideration the isosteric (both canonical and non-canonical) base pairs and multi-pairings in RNA structural motifs. The utility and accuracy of RNAMotifScan is demonstrated by searching for kink-turn, C-loop, sarcin-ricin, reverse kink-turn and E-loop motifs against a 23S rRNA (PDBid: 1S72), which is well characterized for the occurrences of these motifs. Finally, we search these motifs against the RNA structures in the entire Protein Data Bank and the abundances of them are estimated. RNAMotifScan is freely available at our supplementary website (http://genome.ucf.edu/RNAMotifScan)

    Elastic properties of ribosomal RNA building blocks: molecular dynamics of the GTPase-associated center rRNA

    Get PDF
    Explicit solvent molecular dynamics (MD) was used to describe the intrinsic flexibility of the helix 42–44 portion of the 23S rRNA (abbreviated as Kt-42+rGAC; kink-turn 42 and GTPase-associated center rRNA). The bottom part of this molecule consists of alternating rigid and flexible segments. The first flexible segment (Hinge1) is the highly anharmonic kink of Kt-42. The second one (Hinge2) is localized at the junction between helix 42 and helices 43/44. The rigid segments are the two arms of helix 42 flanking the kink. The whole molecule ends up with compact helices 43/44 (Head) which appear to be modestly compressed towards the subunit in the Haloarcula marismortui X-ray structure. Overall, the helix 42–44 rRNA is constructed as a sophisticated intrinsically flexible anisotropic molecular limb. The leading flexibility modes include bending at the hinges and twisting. The Head shows visible internal conformational plasticity, stemming from an intricate set of base pairing patterns including dynamical triads and tetrads. In summary, we demonstrate how rRNA building blocks with contrasting intrinsic flexibilities can form larger architectures with highly specific patterns of preferred low-energy motions and geometries

    Revisiting the planarity of nucleic acid bases: Pyramidilization at glycosidic nitrogen in purine bases is modulated by orientation of glycosidic torsion

    Get PDF
    We describe a novel, fundamental property of nucleobase structure, namely, pyramidilization at the N1/9 sites of purine and pyrimidine bases. Through a combined analyses of ultra-high-resolution X-ray structures of both oligonucleotides extracted from the Nucleic Acid Database and isolated nucleotides and nucleosides from the Cambridge Structural Database, together with a series of quantum chemical calculations, molecular dynamics (MD) simulations, and published solution nuclear magnetic resonance (NMR) data, we show that pyramidilization at the glycosidic nitrogen is an intrinsic property. This property is common to isolated nucleosides and nucleotides as well as oligonucleotides—it is also common to both RNA and DNA. Our analysis suggests that pyramidilization at N1/9 sites depends in a systematic way on the local structure of the nucleoside. Of note, the pyramidilization undergoes stereo-inversion upon reorientation of the glycosidic bond. The extent of the pyramidilization is further modulated by the conformation of the sugar ring. The observed pyramidilization is more pronounced for purine bases, while for pyrimidines it is negligible. We discuss how the assumption of nucleic acid base planarity can lead to systematic errors in determining the conformation of nucleotides from experimental data and from unconstrained MD simulations
    corecore