Repair of Aberrant Splicing in Growth Hormone Receptor by Antisense Oligonucleotides Targeting the Splice Sites of a Pseudoexon

Context: The GH receptor (GHR) pseudoexon 6 Psi defect is a frequent cause of GH insensitivity (GHI) resulting from a non-functioning GH receptor (GHR). It results in a broad range of phenotypes and may also be present in patients diagnosed as idiopathic short stature.Objective: Our objective was to correct aberrant GHR splicing and inclusion of 6 Psi using exon-skipping antisense oligonucleotides (ASOs).Design and Setting: Three ASOs binding the 5' (ASO-5), 3' (ASO-3), and branch site (ASO-Br) of 6 Psi were tested in an in vitro splicing assay and a cell transfection system. The wild-type (wt) and mutant (mt) DNA minigenes (wt- and mtL1-GHR6 Psi-L2, respectively) were created by inserting the GHR 6 Psi in a well-characterized splice reporter (Adml-par). For the in vitro splicing assay, the wt- and mtL1-GHR6 Psi-L2 were transcribed into pre-mRNA in the presence of [alpha P-32]GTP and incubated with ASOs in HeLa nuclear extracts. For the cell transfection studies, wt-and mtL1-GHR6 Psi-L2 cloned into pcDNA 3.1 were transfected with ASOs into HEK293 cells. After 48 h, RNA was extracted and radiolabeled RT-PCR products quantified.Results: ASO-3 induced an almost complete pseudoexon skipping in vitro and in HEK293 cells. This effect was dose dependent and maximal at 125-250 nM. ASO-5 produced modest pseudoexon skipping, whereas ASO-Br had no effect. Targeting of two splice elements simultaneously was less effective than targeting one. ASO-Br was tested on the wtL1-GHR6 Psi-L2 and did not act as an enhancer of 6 Psi inclusion.Conclusions: The exon-skipping ASO approach was effective in correcting aberrant GHR splicing and may be a promising therapeutic tool. (J Clin Endocrinol Metab 95: 3542-3546, 2010

Shock and Release Temperatures in Molybdenum

Shock and release temperatures in Mo were calculated, taking account of heating from plastic flow predicted using the Steinberg-Guinan model. Plastic flow was calculated self-consistently with the shock jump conditions: this is necessary for a rigorous estimate of the locus of shock states accessible. The temperatures obtained were significantly higher than predicted assuming ideal hydrodynamic loading. The temperatures were compared with surface emission spectrometry measurements for Mo shocked to around 60GPa and then released into vacuum or into a LiF window. Shock loading was induced by the impact of a planar projectile, accelerated by high explosive or in a gas gun. Surface velocimetry showed an elastic wave at the start of release from the shocked state; the amplitude of the elastic wave matched the prediction to around 10%, indicating that the predicted flow stress in the shocked state was reasonable. The measured temperatures were consistent with the simulations, indicating that the fraction of plastic work converted to heat was in the range 70-100% for these loading conditions

Explanation for Anomalous Shock Temperatures Measured by Neutron Resonance Spectroscopy

Neutron resonance spectrometry (NRS) has been used to measure the temperature inside Mo samples during shock loading. The temperatures obtained were significantly higher than predicted assuming ideal hydrodynamic loading. The effect of plastic flow and non-ideal projectile behavior were assessed. Plastic flow was calculated self-consistently with the shock jump conditions: this is necessary for a rigorous estimate of the locus of shock states accessible. Plastic flow was estimated to contribute a temperature rise of 53K compared with hydrodynamic flow. Simulations were performed of the operation of the explosively-driven projectile system used to induce the shock in the Mo sample. The simulations predicted that the projectile was significantly curved on impact, and still accelerating. The resulting spatial variations in load, including radial components of velocity, were predicted to increase the apparent temperature that would be deduced from the width of the neutron resonance by 160K. These corrections are sufficient to reconcile the apparent temperatures deduced using NRS with the accepted properties of Mo, in particular its equation of state.Comment: near-final version, waiting for final consent from an autho

Extremely low frequency (0.1 to 1.0 mHz) surf zone currents.

Author Posting. © American Geophysical Union, 2019. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geophysical Research Letters 46(3), (2019):1531-1536, doi:10.1029/2018GL081106.Low‐frequency surf zone eddies disperse material between the shoreline and the continental shelf, and velocity fluctuations with frequencies as low as a few mHz have been observed previously on several beaches. Here spectral estimates of surf zone currents are extended to an order of magnitude lower frequency, resolving an extremely low frequency peak of approximately 0.5 mHz that is observed for a range of beaches and wave conditions. The magnitude of the 0.5‐mHz peak increases with increasing wave energy and with spatial inhomogeneity of bathymetry or currents. The 0.5‐mHz peak may indicate the frequency for which nonlinear energy transfers from higher‐frequency, smaller‐scale motions are balanced by dissipative processes and thus may be the low‐frequency limit of the hypothesized 2‐D cascade of energy from breaking waves to lower frequency motions.We thank R. Guza, T. Herbers, and T. Lippmann for their leadership roles during the SandyDuck and NCEX projects and the CCS (SIO), PVLAB (WHOI), and FRF (USACE) field teams for deploying, maintaining, and recovering sensors in harsh conditions over many years. Funding was provided by ASD(R&E), NSF, and ONR. The data can be obtained via https://chlthredds.erdc.dren.mil/thredds/catalog/frf/catalog.html and https://pv‐lab.org.2019-07-0

Large‐scale hydro‐climatology of the terrestrial Arctic drainage system

The large‐scale hydro‐climatology of the terrestrial Arctic drainage system is examined, focusing on the period 1960 onward. Special attention is paid to the Ob, Yenisey, Lena, and Mackenzie watersheds, which provide the bulk of freshwater discharge to the Arctic Ocean. Station data are used to compile monthly gridded time series of gauge‐corrected precipitation (P). Gridded time series of precipitation minus evapotranspiration (P−ET) are calculated from the moisture flux convergence using NCEP reanalysis data. Estimates of ET are obtained as a residual. Runoff (R) is obtained from available discharge records. For long‐term water‐year means, P−ET for the Yenisey, Lena, and Mackenzie watersheds is 16–20% lower than the observed runoff. In the Ob watershed, the two values agree within 9%. Given the uncertainties in P−ET, we consider the atmospheric and surface water budgets to be reasonably closed. Compared to the other three basins, the mean runoff ratio (R/P) is lower in the Ob watershed, consistent with the high fraction of annual precipitation lost through ET. All basins exhibit summer maxima in P and minima in P−ET. Summer P−ET in the Ob watershed is negative due to high ET rates. For large domains in northern Eurasia, about 25% of July precipitation is associated with the recycling of water vapor evapotranspirated within each domain. This points to a significant effect of the land surface on the hydrologic regime. Variability in P and P−ET has generally clear associations with the regional atmospheric circulation. A strong link with the Urals trough is documented for the Ob. Relationships with indices of the Arctic Oscillation and other teleconnections are generally weak. Water‐year time series of runoff and P−ET are strongly correlated in the Lena watershed only, reflecting extensive permafrost. Cold‐season runoff has increased in the Yenisey and Lena watersheds. This is most pronounced in the Yenisey watershed, where runoff has also increased sharply in spring, decreased in summer, but has increased for the year as a whole. The mechanisms for these changes are not entirely clear. While they fundamentally relate to higher air temperatures, increased winter precipitation, and strong summer drying, we speculate links with changes in active layer thickness and thawing permafrost

Surfzone to inner-shelf exchange estimated from dye tracer balances

Author Posting. © American Geophysical Union, 2015. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research: Oceans 120 (2015): 6289–6308, doi:10.1002/2015JC010844.Surfzone and inner-shelf tracer dispersion are observed at an approximately alongshore-uniform beach. Fluorescent Rhodamine WT dye, released near the shoreline continuously for 6.5 h, is advected alongshore by breaking-wave- and wind-driven currents, and ejected offshore from the surfzone to the inner-shelf by transient rip currents. Novel aerial-based multispectral dye concentration images and in situ measurements of dye, waves, and currents provide tracer transport and dilution observations spanning about 350 m cross-shore and 3 km alongshore. Downstream dilution of near-shoreline dye follows power law decay with exponent −0.33, implying that a tenfold increase in alongshore distance reduces the concentration about 50%. Coupled surfzone and inner-shelf dye mass balances close, and in 5 h, roughly half of the surfzone-released dye is transported offshore to the inner-shelf. Observed cross-shore transports are parameterized well ( inline image, best fit slope inline image) using a bulk exchange velocity and mean surfzone to inner-shelf dye concentration difference. The best fit cross-shore exchange velocity inline image is similar to a temperature-derived exchange velocity on another day with similar wave conditions. The inline image magnitude and observed inner-shelf dye length scales, time scales, and vertical structure indicate the dominance of transient rip currents in surfzone to inner-shelf cross-shore exchange during moderate waves at this alongshore-uniform beach.National Science Foundation Graduate Research Fellowship Grant Number: DGE1144086, California Sea Grant Number: R/CONT-207TR2016-03-1

How today’s shocks predict tomorrow’s leaving

Purpose This research explores “shocking events” as part of the unfolding model of turnover, extending our understanding of the influence of various types of shocks on future voluntary employee separations. Design/Methodology/Approach: 1536 new hires at a large financial institution reported shocks monthly during their first 8 months at work as well as their job satisfaction and perceptions of marketability. We used event history to estimate the basic distributional properties of the shocks and Cox proportional hazards models to examine the effects of shocks on job satisfaction and turnover over the subsequent year as reported by the organization. Findings: Organizational shocks generally occur earlier than personal shocks. Further, unexpected shocks have a stronger impact than expected shocks on subsequent turnover. Finally, the effects of organizational shocks on turnover are mediated by job satisfaction, whereas personal shocks have direct effects on turnover. Implications: Our findings offer evidence for the utility of the shock construct in the unfolding model of turnover and speak to the importance of encouraging managers to monitor shocks on an ongoing basis in order to predict when different types of shocks will occur and their likely influence on turnover. Originality/Value: Ours is the first study to examine shocks as they occur. This is a contrast to prior studies that relied on retrospective accounts. Thus, we are able to test new hypotheses (e.g., direct effects vs. mediation) that expand the unfolding model of turnover

Improving access to psychological therapy: Initial evaluation of two UK demonstration sites

Copyright © 2009 Elsevier. NOTICE: This is the author’s version of a work accepted for publication by Elsevier. Changes resulting from the publishing process, including peer review, editing, corrections, structural formatting and other quality control mechanisms, may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Behaviour Research and Therapy, 2009, Vol. 47, Issue 11, pp. 910 – 920 DOI: http://dx.doi.org/10.1016/j.brat.2009.07.010Recently the UK Government announced an unprecedented, large-scale initiative for Improving Access to Psychological Therapies (IAPT) for depression and anxiety disorders. Prior to this development, the Department of Health established two pilot projects that aimed to collect valuable information to inform the national roll-out. Doncaster and Newham received additional funds to rapidly increase the availability of CBT-related interventions and to deploy them in new clinical services, operating on stepped-care principles, when appropriate. This article reports an evaluation of the new services (termed ‘demonstration sites’) during their first thirteen months of operation. A session-by-session outcome monitoring system achieved unusually high levels of pre to post-treatment data completeness. Large numbers of patients were treated, with low-intensity interventions (such as guided self-help) being particularly helpful for achieving high throughput. Clinical outcomes were broadly in line with expectation. 55–56% of patients who had attended at least twice (including the assessment interview) were classified as recovered when they left the services and 5% had improved their employment status. Treatment gains were largely maintained at 10 month follow-up. Opening the services to self-referral appeared to facilitate access for some groups that tend to be underrepresented in general practice referrals. Outcomes were comparable for the different ethnic groups who access the services. Issues for the further development of IAPT are discussed

Unsolicited written narratives as a methodological genre in terminal illness: challenges and limitations

Stories about illness have proven invaluable in helping health professionals understand illness experiences. Such narratives have traditionally been solicited by researchers through interviews and the collection of personal writings, including diaries. These approaches are, however, researcher driven; the impetus for the creation of the story comes from the researcher and not the narrator. In recent years there has been exponential growth in illness narratives created by individuals, of their own volition, and made available for others to read in print or as Internet accounts. We sought to determine whether it was possible to identify such material for use as research data to explore the subject of living with the terminal illness amyotrophic lateral sclerosis/motor neuron disease—the contention being that these accounts are narrator driven and therefore focus on issues of greatest importance to the affected person. We encountered and sought to overcome a number of methodological and ethical challenges, which is our focus here