Triple-helix formation in the antiparallel binding motif of oligodeoxynucleotides containing N(9)- and N(7)-2-aminopurine deoxynucleosides

Triplex-forming oligodeoxynucleotide 15mers, designed to bind in the antiparallel triple-helical binding motif, containing single substitutions (Z) of the four isomeric alphaN(7)-, betaN(7)-, alphaN(9)- and betaN(9)-2-aminopurine (ap)-deoxyribonucleosides were prepared. Their association with double-stranded DNA targets containing all four natural base pairs (X-Y) opposite the aminopurine residues was determined by quantitative DNase I footprint titration in the absence of monovalent metal cations. The corresponding association constants were found to be in a rather narrow range between 1.0 x 10(6) and 1.3 x 10(8) M(-1). The following relative order in Z x X-Y base-triple stabilities was found: Z = alphaN(7)ap: T-A > A-T> C-G approximately G-C; Z = betaN(7)ap: A-T > C-G > G-C > T-A; Z = alphaN(9)ap: A-T = G-C > T-A > C-G; and Z = betaN(9)ap: G-C > A-T > C-G > T-A

Genetic variation in IL28B is associated with chronic hepatitis C and treatment failure: a genome-wide association study

Hepatitis C virus (HCV) induces chronic infection in 50% to 80% of infected persons; approximately 50% of these do not respond to therapy. We performed a genome-wide association study to screen for host genetic determinants of HCV persistence and response to therapy.

Transport model of the human Na+-coupled L-ascorbic acid (vitamin C) transporter SVCT1

Vitamin C (L-ascorbic acid) is an essential micronutrient that serves as an antioxidant and as a cofactor in many enzymatic reactions. Intestinal absorption and renal reabsorption of the vitamin is mediated by the epithelial apical L-ascorbic acid cotransporter SVCT1 (SLC23A1). We explored the molecular mechanisms of SVCT1-mediated L-ascorbic acid transport using radiotracer and voltage-clamp techniques in RNA-injected Xenopus oocytes. L-ascorbic acid transport was saturable (K(0.5) approximately 70 microM), temperature dependent (Q(10) approximately 5), and energized by the Na(+) electrochemical potential gradient. We obtained a Na(+)-L-ascorbic acid coupling ratio of 2:1 from simultaneous measurement of currents and fluxes. L-ascorbic acid and Na(+) saturation kinetics as a function of cosubstrate concentrations revealed a simultaneous transport mechanism in which binding is ordered Na(+), L-ascorbic acid, Na(+). In the absence of L-ascorbic acid, SVCT1 mediated pre-steady-state currents that decayed with time constants 3-15 ms. Transients were described by single Boltzmann distributions. At 100 mM Na(+), maximal charge translocation (Q(max)) was approximately 25 nC, around a midpoint (V(0.5)) at -9 mV, and with apparent valence approximately -1. Q(max) was conserved upon progressive removal of Na(+), whereas V(0.5) shifted to more hyperpolarized potentials. Model simulation predicted that the pre-steady-state current predominantly results from an ion-well effect on binding of the first Na(+) partway within the membrane electric field. We present a transport model for SVCT1 that will provide a framework for investigating the impact of specific mutations and polymorphisms in SLC23A1 and help us better understand the contribution of SVCT1 to vitamin C metabolism in health and disease.

Cardiac c-kit+AT2+ cell population is increased in response to ischemic injury and supports cardiomyocyte performance

The expression pattern of angiotensin AT2 receptors with predominance during fetal life and upregulation under pathological conditions during tissue injury/repair process suggests that AT2 receptors may exert an important action in injury/repair adaptive mechanisms. Less is known about AT2 receptors in acute ischemia-induced cardiac injury. We aimed here to elucidate the role of AT2 receptors after acute myocardial infarction. Double immunofluorescence staining showed that cardiac AT2 receptors were mainly detected in clusters of small c-kit+ cells accumulating in peri-infarct zone and c-kit+AT2+ cells increased in response to acute cardiac injury. Further, we isolated cardiac c-kit+AT2+ cell population by modified magnetic activated cell sorting and fluorescence activated cell sorting. These cardiac c-kit+AT2+ cells, represented approximately 0.19% of total cardiac cells in infarcted heart, were characterized by upregulated transcription factors implicated in cardiogenic differentiation (Gata-4, Notch-2, Nkx-2.5) and genes required for self-renewal (Tbx-3, c-Myc, Akt). When adult cardiomyocytes and cardiac c-kit+AT2+ cells isolated from infarcted rat hearts were cocultured, AT2 receptor stimulation in vitro inhibited apoptosis of these cocultured cardiomyocytes. Moreover, in vivo AT2 receptor stimulation led to an increased c-kit+AT2+ cell population in the infarcted myocardium and reduced apoptosis of cardiomyocytes in rats with acute myocardial infarction. These data suggest that cardiac c-kit+AT2+ cell population exists and increases after acute ischemic injury. AT2 receptor activation supports performance of cardiomyocytes, thus contributing to cardioprotection via cardiac c-kit+AT2+ cell population.

The Southern Hemisphere at glacial terminations: insights from the Dome C ice core

The many different proxy records from the European Project for Ice Coring in Antarctica (EPICA) Dome C ice core allow for the first time a comparison of nine glacial terminations in great detail. Despite the fact that all terminations cover the transition from a glacial maximum into an interglacial, there are large differences between single terminations. For some terminations, Antarctic temperature increased only moderately, while for others, the amplitude of change at the termination was much larger. For the different terminations, the rate of change in temperature is more similar than the magnitude or duration of change. These temperature changes were accompanied by vast changes in dust and sea salt deposition all over Antarctica. Here we investigate the phasing between a South American dust proxy (non-sea-salt calcium flux, nssCa2+), a sea ice proxy (sea salt sodium flux, ssNa+) and a proxy for Antarctic temperature (deuterium, δD). In particular, we look into whether a similar sequence of events applies to all terminations, despite their different characteristics. All proxies are derived from the EPICA Dome C ice core, resulting in a relative dating uncertainty between the proxies of less than 20 years. At the start of the terminations, the temperature (δD) increase and dust (nssCa2+ flux) decrease start synchronously. The sea ice proxy (ssNa+ flux), however, only changes once the temperature has reached a particular threshold, approximately 5°C below present day temperatures (corresponding to a δD value of −420‰). This reflects to a large extent the limited sensitivity of the sea ice proxy during very cold periods with large sea ice extent. At terminations where this threshold is not reached (TVI, TVIII), ssNa+ flux shows no changes. Above this threshold, the sea ice proxy is closely coupled to the Antarctic temperature, and interglacial levels are reached at the same time for both ssNa+ and δD. On the other hand, once another threshold at approximately 2°C below present day temperature is passed (corresponding to a δD value of −402‰), nssCa2+ flux has reached interglacial levels and does not change any more, despite further warming. This threshold behaviour most likely results from a combination of changes to the threshold friction velocity for dust entrainment and to the distribution of surface wind speeds in the dust source region.

Validation of middle-atmospheric campaign-based water vapour measured by the ground-based microwave radiometer MIAWARA-C

Middle atmospheric water vapour can be used as a tracer for dynamical processes. It is mainly measured by satellite instruments and ground-based microwave radiometers. Ground-based instruments capable of measuring middle-atmospheric water vapour are sparse but valuable as they complement satellite measurements, are relatively easy to maintain and have a long lifetime. MIAWARA-C is a ground-based microwave radiometer for middle-atmospheric water vapour designed for use on measurement campaigns for both atmospheric case studies and instrument intercomparisons. MIAWARA-C's retrieval version 1.1 (v1.1) is set up in a such way as to provide a consistent data set even if the instrument is operated from different locations on a campaign basis. The sensitive altitude range for v1.1 extends from 4 hPa (37 km) to 0.017 hPa (75 km). For v1.1 the estimated systematic error is approximately 10% for all altitudes. At lower altitudes it is dominated by uncertainties in the calibration, with altitude the influence of spectroscopic and temperature uncertainties increases. The estimated random error increases with altitude from 5 to 25%. MIAWARA-C measures two polarisations of the incident radiation in separate receiver channels, and can therefore provide two measurements of the same air mass with independent instrumental noise. The standard deviation of the difference between the profiles obtained from the two polarisations is in excellent agreement with the estimated random measurement error of v1.1. In this paper, the quality of v1.1 data is assessed for measurements obtained at two different locations: (1) a total of 25 months of measurements in the Arctic (Sodankylä, 67.37° N, 26.63° E) and (2) nine months of measurements at mid-latitudes (Zimmerwald, 46.88° N, 7.46° E). For both locations MIAWARA-C's profiles are compared to measurements from the satellite experiments Aura MLS and MIPAS. In addition, comparisons to ACE-FTS and SOFIE are presented for the Arctic and to the ground-based radiometer MIAWARA for the mid-latitude campaigns. In general, all intercomparisons show high correlation coefficients, confirming the ability of MIAWARA-C to monitor temporal variations of the order of days. The biases are generally below 13% and within the estimated systematic uncertainty of MIAWARA-C. No consistent wet or dry bias is identified for MIAWARA-C. In addition, comparisons to the reference instruments indicate the estimated random error of v1.1 to be a realistic measure of the random variation on the retrieved profile between 45 and 70 km.

Hepatitis C: a changing epidemic.

Approximately 3% of the world population is estimated to have a chronic hepatitis C virus (HCV) infection and 500,000 individuals die from its consequences yearly. Persons who inject drugs (PWID) bear the majority of the disease burden in high-income countries. Drug substitution programmes have helped reduce HCV transmissions among PWID. However, recent epidemics of sexually transmitted HCV infections in HIV-infected men who have sex with men demonstrated the changing nature of the HCV epidemic. HCV therapy is undergoing a revolution, as new interferon-free, oral treatments eradicate HCV infections in almost all treated patients. As a consequence, the eradication of HCV has become a matter of debate and is becoming an important future public health target. However, for this to be achieved, many challenges need to be addressed, including the poor uptake of HCV testing, the high cost of the new antiviral combinations and the high frequency of re-infections after treatment in some populations.