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

One service, many voices: enhancing consumer participation in a primary health service for multicultural women

Background: Consumer participation in primary health care is important in providing quality consumer-focused care but challenging when working with disadvantaged groups of diverse cultural and linguistic backgrounds. Women's Health Services (WHS) works with women from over sixty different nationalities, including many newly arrived migrants and refugees. New arrivals access a wide range of WHS programs including medical services, counselling, information, community talks and workshops, referral, and outreach, but few ethnic women attended the alcohol and other drug (AOD) services offered by the organisation.Aim: To establish an active consumer reference group to assist understanding and reducing the barriers to AOD services for a heterogeneous disadvantaged group that includes individuals from different cultural, language and educational backgrounds. Results: Leaning heavily on experiences from the mental health field, WHS overcame many practical and philosophical considerations which included: agreeing upon the purpose of the group and how it would operate within the structure of the organisation; the level of English language required by participants for the group to function; issues of resourcing the group; and ensuring an appropriate, workable demographic mix in terms of age, language, and migration experiences. Conclusion: The process and the outcome of establishing a Consumer Reference Group (CRG) in a primary health care setting has been valuable for consumers and health service providers within the organisation

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

Incidental Findings on CT Scans in the Emergency Department

Objectives. Incidental findings on computed tomography (CT) scans are common. We sought to examine rates of findings and disclosure among discharged patients who received a CT scan in the ED. Methods. Retrospective chart review (Aug-Oct 2009) of 600 patients age 18 and older discharged home from an urban Level 1 trauma center. CT reports were used to identify incidental findings and discharge paperwork was used to determine whether the patient was informed of these findings. Results. There were 682 CT scans among 600 patients: 199 Abdomen & Pelvis, 405 Head, and 78 Thorax. A total of 348 incidental findings were documented in 228/682 (33.4%) of the scans, of which 34 (9.8%) were reported to patients in discharge paperwork. Patients with 1 incidental finding were less likely to receive disclosure than patients with 2 or more (P = .010). Patients age <60 were less likely to have incidental findings (P < .001). There was no significant disclosure or incidental finding difference by gender. Conclusions. While previous research suggests that CT incidental findings are often benign, reporting to patients is recommended but this is rarely happening

The double-edged sword: Emotional regulation for children at risk.

Abstract: The capacity to manage emotion is based on the growth of self-regulatory capacities in the early years, but is also affected by situational demands, influences from other people, and the child&apos;s goals for regulating emotion in a particular setting. For most children growing up in supportive contexts, the growth of emotional regulation is associated with enhanced psychosocial wellbeing and socioemotional competence. But for children who are at risk for the development of psychopathology owing to environmental stresses or intrinsic vulnerability (or their interaction), emotional regulation often entails inherent trade-offs that make nonoptimal strategies of managing emotion expectable, perhaps inevitable, in a context of difficult environmental demands and conflicting emotional goals. This analysis discusses how emotional regulation in children at risk may simultaneously foster both resiliency and vulnerability by considering how emotion is managed when children (a) are living with a parent who is depressed, (b) witness or experience domestic violence, or (c) are temperamentally inhibited when encountering novel challenges. In each case, the child,s efforts to manage emotion may simultaneously buffer against certain stresses while also enhancing the child&apos;s vulnerability to other risks and demands. This double-edged sword of emotional regulation in conditions of risk for children cautions against using &quot;optimal&quot; emotional regulation as an evaluative standard for such children or assuming that emotional regulation necessarily improves psychosocial well-being. It also suggests how the study of emotional regulation must consider the goals for regulating emotion and the contexts in which those goals are sought. Article: Emotions are complex phenomena. They entail constellations of physiological arousal, cognitive appraisal, and expressive activity that interact with situational demands and cultural rules to create subtly nuanced, richly variable subjective and behavioral events. Individual differences in emotion are also multifaceted, fashioned from the interaction of organismic (e.g., temperamental) characteristics, experiential history, construals of oneself and the situation, and the personal goals that shape emotional arousal and its expression. Consequently, although emotion has its roots in the legacy of biological adaptation, it also reflects some of the most sophisticated features of human social cognition, self-understanding, and strategic functioning. It should be no surprise, therefore, that emotional regulation is a complex phenomenon. Although strategies of emotional self-regulation originate in the young infant&apos;s simple efforts to cope with distress through self-soothing, they quickly become integrated into a network o

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

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

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