Nucleic acid structure and sequence probing using fluorescent base analogue tC^o

The fluorescent cytosine analog tC(o) is on average the brightest probe of its kind and, moreover, it introduces minimal perturbations to the normal secondary structure of DNA. Here several ways of how tC(o), with an advantage, can be used as a local fluorescent probe in nucleic acid systems are presented. Most importantly, we show that tCo is an excellent probe for the detection of individual melting processes of complex nucleic acid structures containing a large number of separate secondary structure motifs. Since conventional UV-melting investigations merely monitor the global melting process of the whole nucleic acid structure, e.g. multi-hairpin systems in RNA/DNA, and thus is incapable of estimating individual melting transitions of such systems, tC(o) represents a new method of characterization. Furthermore, we find that tCo may be used to detect bulges and loops in nucleic acids as well as to distinguish a matched base-pair from several of the mismatched

The central dynamics of M3, M13, and M92: Stringent limits on the masses of intermediate-mass black holes

We used the PMAS integral field spectrograph to obtain large sets of radial velocities in the central regions of three northern Galactic globular clusters: M3, M13, and M92. By applying the novel technique of crowded field 3D spectroscopy, we measured radial velocities for about 80 stars within the central ~ 10 arcsec of each cluster. These are by far the largest spectroscopic datasets obtained in the innermost parts of these clusters up to now. To obtain kinematical data across the whole extent of the clusters, we complement our data with measurements available in the literature. We combine our velocity measurements with surface brightness profiles to analyse the internal dynamics of each cluster using spherical Jeans models, and investigate whether our data provide evidence for an intermediate-mass black hole in any of the clusters. The surface brightness profiles reveal that all three clusters are consistent with a core profile, although shallow cusps cannot be excluded. We find that spherical Jeans models with a constant mass-to-light ratio provide a good overall representation of the kinematical data. A massive black hole is required in none of the three clusters to explain the observed kinematics. Our 1sigma (3sigma) upper limits are 5300 M_sun (12000 M_sun) for M3, 8600 M_sun (13000 M_sun) for M13, and 980 M_sun (2700 M_sun) for M92. A puzzling circumstance is the existence of several potential high velocity stars in M3 and M13, as their presence can account for the majority of the discrepancies that we find in our mass limits compared to M92.Comment: accepted for publication in A&A, 20 pages, 15 figures, tables D1 to D6 only available at CD

Towards DIB mapping in galaxies beyond 100 Mpc. A radial profile of the λ\lambda5780.5 diffuse interstellar band in AM 1353-272 B

Diffuse Interstellar Bands (DIBs) are non-stellar weak absorption features of unknown origin found in the spectra of stars viewed through one or several clouds of Interstellar Medium (ISM). Research of DIBs outside the Milky Way is currently very limited. Specifically spatially resolved investigations of DIBs outside of the Local Group is, to our knowledge, inexistent. Here, we explore the capability of the high sensitivity Integral Field Spectrograph, MUSE, as a tool to map diffuse interstellar bands at distances larger than 100 Mpc. We use MUSE commissioning data for AM 1353-272 B, the member with highest extinction of the "The Dentist's Chair", an interacting system of two spiral galaxies. High signal-to-noise spectra were created by co-adding the signal of many spatial elements distributed in a geometry of concentric elliptical half-rings. We derived decreasing radial profiles for the equivalent width of the $\lambda$5780.5 DIB both in the receding and approaching side of the companion galaxy up to distances of $\sim$4.6 kpc from the center of the galaxy. Likewise, interstellar extinction, as derived from the Halpha/Hbeta line ratio displays a similar trend, with decreasing values towards the external parts. This translates into an intrinsic correlation between the strength of the DIB and the extinction within AM 1353-272 B consistent with the current existing global trend between these quantities when using measurements for both Galactic and extragalactic sight lines. Mapping of DIB strength in the Local Universe as up to now only done for the Milky Way seems feasible. This offers a new approach to study the relationship between DIBs and other characteristics and species of the ISM in different conditions as those found in our Galaxy to the use of galaxies in the Local Group and/or single sightlines towards supernovae, quasars and galaxies outside the Local Group.Comment: 4 pages, 4 figures, accepted for publication as a Letter in Astronomy and Astrophysics; Received 10 February 2015 / Accepted 20 February 2015 ; English corrections include

Highly efficient incorporation of the fluorescent nucleotide analogs tC and tCO by Klenow fragment

Studies of the mechanisms by which DNA polymerases select the correct nucleotide frequently employ fluorescently labeled DNA to monitor conformational rearrangements of the polymerase–DNA complex in response to incoming nucleotides. For this purpose, fluorescent base analogs play an increasingly important role because they interfere less with the DNA–protein interaction than do tethered fluorophores. Here we report the incorporation of the 5′-triphosphates of two exceptionally bright cytosine analogs, 1,3-diaza-2-oxo-phenothiazine (tC) and its oxo-homolog, 1,3-diaza-2-oxo-phenoxazine (tCO), into DNA by the Klenow fragment. Both nucleotide analogs are polymerized with slightly higher efficiency opposite guanine than cytosine triphosphate and are shown to bind with nanomolar affinity to the DNA polymerase active site, according to fluorescence anisotropy measurements. Using this method, we perform competitive binding experiments and show that they can be used to determine the dissociation constant of any given natural or unnatural nucleotide. The results demonstrate that the active site of the Klenow fragment is flexible enough to tolerate base pairs that are size-expanded in the major groove. In addition, the possibility to enzymatically polymerize a fluorescent nucleotide with high efficiency complements the tool box of biophysical probes available to study DNA replication

Определение сроков годности экстракционных лекарственных средств корневищ с корнями синюхи голубой

РАСТЕНИЙ ЭКСТРАКТЫСИНЮХА ГОЛУБА

Plasma lipid composition and risk of developing cardiovascular disease.

We tested whether characteristic changes of the plasma lipidome in individuals with comparable total lipids level associate with future cardiovascular disease (CVD) outcome and whether 23 validated gene variants associated with coronary artery disease (CAD) affect CVD associated lipid species

A MUSE map of the central Orion Nebula (M 42)

We present a new integral-field spectroscopic dataset of the central part of the Orion Nebula (M 42), observed with the MUSE instrument at the ESO VLT. We reduced the data with the public MUSE pipeline. The output products are two FITS cubes with a spatial size of ~5.9'x4.9' (corresponding to ~0.76 pc x 0.63 pc) and a contiguous wavelength coverage of 4595...9366 Angstrom, spatially sampled at 0.2". We provide two versions with a sampling of 1.25 Angstrom and 0.85 Angstrom in dispersion direction. Together with variance cubes these files have a size of 75 and 110 GiB on disk. They represent one of the largest integral field mosaics to date in terms of information content. We make them available for use in the community. To validate this dataset, we compare world coordinates, reconstructed magnitudes, velocities, and absolute and relative emission line fluxes to the literature and find excellent agreement. We derive a two-dimensional map of extinction and present de-reddened flux maps of several individual emission lines and of diagnostic line ratios. We estimate physical properties of the Orion Nebula, using the emission line ratios [N II] and [S III] (for the electron temperature $T_e$) and [S II] and [Cl III] (for the electron density $N_e$), and show two-dimensional images of the velocity measured from several bright emission lines.Comment: Resubmitted to A&A after incorporating referee comments; access to full dataset via http://muse-vlt.eu/science/data-release

Characterization of photophysical and base-mimicking properties of a novel fluorescent adenine analogue in DNA

To increase the diversity of fluorescent base analogues with improved properties, we here present the straightforward click-chemistry-based synthesis of a novel fluorescent adenine-analogue triazole adenine (AT) and its photophysical characterization inside DNA. AT shows promising properties compared to the widely used adenine analogue 2-aminopurine. Quantum yields reach >20% and >5% in single- and double-stranded DNA, respectively, and show dependence on neighbouring bases. Moreover, AT shows only a minor destabilization of DNA duplexes, comparable to 2-aminopurine, and circular dichroism investigations suggest that AT only causes minimal structural perturbations to normal B-DNA. Furthermore, we find that AT shows favourable base-pairing properties with thymine and more surprisingly also with normal adenine. In conclusion, AT shows strong potential as a new fluorescent adenine analogue for monitoring changes within its microenvironment in DNA