Fluorescence Correlation Spectroscopy Reveals Efficient Cytosolic Delivery of Protein Cargo by Cell-Permeant Miniature Proteins.

New methods for delivering proteins into the cytosol of mammalian cells are being reported at a rapid pace. Differentiating between these methods in a quantitative manner is difficult, however, as most assays for evaluating cytosolic protein delivery are qualitative and indirect and thus often misleading. Here we make use of fluorescence correlation spectroscopy (FCS) to determine with precision and accuracy the relative efficiencies with which seven different previously reported "cell-penetrating peptides" (CPPs) transport a model protein cargo-the self-labeling enzyme SNAP-tag-beyond endosomal membranes and into the cytosol. Using FCS, we discovered that the miniature protein ZF5.3 is an exceptional vehicle for delivering SNAP-tag to the cytosol. When delivered by ZF5.3, SNAP-tag can achieve a cytosolic concentration as high as 250 nM, generally at least 2-fold and as much as 6-fold higher than any other CPP evaluated. Additionally, we show that ZF5.3 can be fused to a second enzyme cargo-the engineered peroxidase APEX2-and reliably delivers the active enzyme to the cell interior. As FCS allows one to realistically assess the relative merits of protein transduction domains, we anticipate that it will greatly accelerate the identification, evaluation, and optimization of strategies to deliver large, intact proteins to intracellular locales

Limits On Planets Around Pulsating White Dwarf Stars

We present limits on planetary companions to pulsating white dwarf stars. A subset of these stars exhibit extreme stability in the period and phase of some of their pulsation modes; a planet can be detected around such a star by searching for periodic variations in the arrival time of these pulsations. We present limits on companions greater than a few Jupiter masses around a sample of 15 white dwarf stars as part of an ongoing survey. One star shows a variation in arrival time consistent with a 2M(J) planet in a 4.5 yr orbit. We discuss other possible explanations for the observed signal and conclude that a planet is the most plausible explanation based on the data available.NASA Origins NAG5-13094Astronom

Evidence For Temperature Change And Oblique Pulsation From Light Curve Fits Of The Pulsating White Dwarf GD 358

Convective driving, the mechanism originally proposed by Brickhill for pulsating white dwarf stars, has gained general acceptance as the generic linear instability mechanism in DAV and dbV white dwarfs. This physical mechanism naturally leads to a nonlinear formulation, reproducing the observed light curves of many pulsating white dwarfs. This numerical model can also provide information on the average depth of a star's convection zone and the inclination angle of its pulsation axis. In this paper, we give two sets of results of nonlinear light curve fits to data on the dbV GD 358. Our first fit is based on data gathered in 2006 by the Whole Earth Telescope; this data set was multiperiodic containing at least 12 individual modes. Our second fit utilizes data obtained in 1996, when GD 358 underwent a dramatic change in excited frequencies accompanied by a rapid increase in fractional amplitude; during this event it was essentially monoperiodic. We argue that GD 358's convection zone was much thinner in 1996 than in 2006, and we interpret this as a result of a short-lived increase in its surface temperature. In addition, we find strong evidence of oblique pulsation using two sets of evenly split triplets in the 2006 data. This marks the first time that oblique pulsation has been identified in a variable white dwarf star.Delaware Asteroseismic Research CenterNational Science Foundation AST-0909107, AST-0607840Norman Hackerman Advanced Research Program 003658-0255-2007Crystal Trust FoundationMt. Cuba ObservatoryUniversity of DelawareAstronom

A Class of Eccentric Binaries with Dynamic Tidal Distortions Discovered with Kepler

We have discovered a class of eccentric binary systems within the Kepler data archive that have dynamic tidal distortions and tidally-induced pulsations. Each has a uniquely shaped light curve that is characterized by periodic brightening or variability at time scales of 4-20 days, frequently accompanied by shorter period oscillations. We can explain the dominant features of the entire class with orbitally-varying tidal forces that occur in close, eccentric binary systems. The large variety of light curve shapes arises from viewing systems at different angles. This hypothesis is supported by spectroscopic radial velocity measurements for five systems, each showing evidence of being in an eccentric binary system. Prior to the discovery of these 17 new systems, only four stars, where KOI-54 is the best example, were known to have evidence of these dynamic tides and tidally-induced oscillations. We perform preliminary fits to the light curves and radial velocity data, present the overall properties of this class and discuss the work required to accurately model these systems.Comment: 13 pages, submitted to Ap

Measuring Transit Signal Recovery in the Kepler Pipeline II: Detection Efficiency as Calculated in One Year of Data

The Kepler planet sample can only be used to reconstruct the underlying planet occurrence rate if the detection efficiency of the Kepler pipeline is known, here we present the results of a second experiment aimed at characterising this detection efficiency. We inject simulated transiting planet signals into the pixel data of ~10,000 targets, spanning one year of observations, and process the pixels as normal. We compare the set of detections made by the pipeline with the expectation from the set of simulated planets, and construct a sensitivity curve of signal recovery as a function of the signal-to-noise of the simulated transit signal train. The sensitivity curve does not meet the hypothetical maximum detection efficiency, however it is not as pessimistic as some of the published estimates of the detection efficiency. For the FGK stars in our sample, the sensitivity curve is well fit by a gamma function with the coefficients a = 4.35 and b = 1.05. We also find that the pipeline algorithms recover the depths and periods of the injected signals with very high fidelity, especially for periods longer than 10 days. We perform a simplified occurrence rate calculation using the measured detection efficiency compared to previous assumptions of the detection efficiency found in the literature to demonstrate the systematic error introduced into the resulting occurrence rates. The discrepancies in the calculated occurrence rates may go some way towards reconciling some of the inconsistencies found in the literature.Comment: 13 pages, 7 figures, 1 electronic table, accepted by Ap

Evaluation of MERRA-2-Based Ozone Profile Simulations with the Global Ozonesonde Network

Chemical transport model (CTM) hindcasts of ozone (O3) are useful for filling in observational gaps and providing context for observed O3 variability and trends. We use global networks of ozonesonde stations to evaluate the O3 profiles in two simulations running versions of the NASA Global Modeling Initiative (GMI) chemical mechanism. Both simulations are tied to the NASA Modern-Era Retrospective analysis for Research and Applications, Version 2 (MERRA-2) meteorological reanalysis: 1) The GMI CTM, and 2) The MERRA-2 GMI Replay (M2 GMI). Both simulations start in 1980, and are compared against >50,000 ozonesonde profiles from 37 global stations from the tropics to the poles. The comparisons allow us to evaluate how the Replay technique affects modeled O3 distribution, how an updated chemical mechanism in the GMI CTM affects simulated tropospheric O3 amounts, and how observed O3 distributions compare to the full set of model output. In general, M2 GMI O3 is ~10% higher than in the GMI CTM, and shows global near-surface and tropical upper troposphere/lower stratosphere (UT/LS) high biases. The updated chemical mechanism in the GMI CTM reduces these high biases. Both simulations show similar negative biases in tropical free-tropospheric O3, especially during typical biomass burning seasons. The simulations are highly-correlated with ozonesonde measurements, particularly in the UT/LS (r > 0.8), showing the ability of MERRA-2 to capture tropopause height variations. Both simulations show improved correlations with ozonesonde data and smaller O3 biases in recent years. We expect to use the sonde/model comparisons to diagnose causes of disagreement and to gauge the feasibility of calculating multidecadal O3 trends from the model output

Resting-state anticorrelations between medial and lateral prefrontal cortex: Association with working memory, aging, and individual differences

We examined how variation in working memory (WM) capacity due to aging or individual differences among young adults is associated with intrinsic or resting-state anticorrelations, particularly between (1) the medial prefrontal cortex (MPFC), a component of the default-mode network (DMN) that typically decreases in activation during external, attention-demanding tasks, and (2) the dorsolateral prefrontal cortex (DLPFC), a component of the fronto-parietal control network that supports executive functions and WM and typically increases in activation during attention-demanding tasks. We compared the magnitudes of MPFC-DLPFC anticorrelations between healthy younger and older participants (Experiment 1) and related the magnitudes of these anticorrelations to individual differences on two behavioral measures of WM capacity in two independent groups of young adults (Experiments 1 and 2). Relative to younger adults, older adults exhibited reductions in WM capacity and in MPFC-DLPFC anticorrelations. Within younger adults, greater MPFC-DLPFC anticorrelation at rest correlated with greater WM capacity. These findings show that variation in MPFC-DLPFC anticorrelations, whether related to aging or to individual differences, may reflect an intrinsic functional brain architecture supportive of WM capacity.National Institutes of Health (U.S.) (National Institute on Aging Grant R21 AG030770)National Institutes of Health (U.S.) (Grant T32 GM007484)Barbara J. Weedon Fund Fellowshi

High Resolution Spectroscopy of the Pulsating White Dwarf G29-38

We present the analysis of time-resolved, high resolution spectra of the cool white dwarf pulsator, G29-38. From measuring the Doppler shifts of the H-alpha core, we detect velocity changes as large as 16.5 km/s and conclude that they are due to the horizontal motions associated with the g-mode pulsations on the star. We detect seven pulsation modes from the velocity time-series and identify the same modes in the flux variations. We discuss the properties of these modes and use the advantage of having both velocity and flux measurements of the pulsations to test the convective driving theory proposed for DAV stars. Our data show limited agreement with the expected relationships between the amplitude and phases of the velocity and flux modes. Unexpectedly, the velocity curve shows evidence for harmonic distortion, in the form of a peak in the Fourier transform whose frequency is the exact sum of the two largest frequencies. Combination frequencies are a characteristic feature of the Fourier transforms of light curves of G29-38, but before now have not been detected in the velocities, nor does published theory predict that they should exist. We compare our velocity combination frequency to combination frequencies found in the analysis of light curves of G29-38, and discuss what might account for the existence of velocity combinations with the properties we observe. We also use our high-resolution spectra to determine if either rotation or pulsation can explain the truncated shape observed for the DAV star's line core. We are able to eliminate both mechanisms: the average spectrum does not fit the rotationally broadened model and the time-series of spectra provides proof that the pulsations do not significantly truncate the line.Comment: 24 pages, 9 figures, Accepted for publication in ApJ (June

Revisiting Ruddick: Feminism, pacifism and non-violence

This article explores feminist contentions over pacifism and non-violence in the contextof the Greenham Common Peace Camp in the 1980s and later developments offeminist Just War Theory. We argue that Sara Ruddick’s work puts feminist pacifism, its radical feminist critics and feminist just war theory equally into question. Although Ruddick does not resolve the contestations within feminism over peace, violence and the questions of war, she offers a productive way of holding the tension between them. In our judgment, her work is helpful not only for developing a feminist political response to the threats and temptations of violent strategies but also for thinking through the question of the relation between violence and politics as such