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

Flight Test Results for UAVs Using Boid Guidance Algorithms

A critical technology for operating groups of Uninhabited Aerial Vehicles (UAVs) is distributed guidance. Distributed guidance allows an operator to command several vehicles at the same time, reduces operator workload, and adds redundancy to the system. Some of the leading software candidates for achieving distributed guidance are known as Boid Guidance Algorithms (BGAs), which are agent-based techniques relying on the interactions of simple behaviors. Flight tests are crucial to the advancement of flight technologies such as BGAs, and this was identified as an important area for development. This paper presents the results from the 2005 flight tests of BGAs at NASA Dryden Flight Research Center with two RnR Products’ APV-3 UAVs employing CloudCap Technology\u27s Piccolo autopilot system. Major challenges in these flight tests include the use of a waypoint-following system, limited computation resources, and management of safety procedures. The conclusions of this work include the need for using a path-following platform and completion of a full system optimization. This work is an important step in the development of a deployable distributed guidance system

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

Vorticity generation by short-crested wave breaking

Author Posting. © American Geophysical Union, 2012. 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 39 (2012): L24604, doi:10.1029/2012GL054034.Eddies and vortices associated with breaking waves rapidly disperse pollution, nutrients, and terrestrial material along the coast. Although theory and numerical models suggest that vorticity is generated near the ends of a breaking wave crest, this hypothesis has not been tested in the field. Here we report the first observations of wave-generated vertical vorticity (e.g., horizontal eddies), and find that individual short-crested breaking waves generate significant vorticity [O(0.01 s−1)] in the surfzone. Left- and right-handed wave ends generate vorticity of opposite sign, consistent with theory. In contrast to theory, the observed vorticity also increases inside the breaking crest, possibly owing to onshore advection of vorticity generated at previous stages of breaking or from the shape of the breaking region. Short-crested breaking transferred energy from incident waves to lower frequency rotational motions that are a primary mechanism for dispersion near the shoreline.Funding was provided by a National Security Science and Engineering Faculty Fellowship, the Office of Naval Research, and a Woods Hole Oceanographic Institution Postdoctoral Fellowship.2013-06-2

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

Modeling surf zone tracer plumes : 2. Transport and dispersion

Author Posting. © American Geophysical Union, 2011. 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 116 (2011): C11028, doi:10.1029/2011JC007211.Five surf zone dye tracer releases from the HB06 experiment are simulated with a tracer advection diffusion model coupled to a Boussinesq surf zone model (funwaveC). Model tracer is transported and stirred by currents and eddies and diffused with a breaking wave eddy diffusivity, set equal to the breaking wave eddy viscosity, and a small (0.01 m2 s−1) background diffusivity. Observed and modeled alongshore parallel tracer plumes, transported by the wave driven alongshore current, have qualitatively similar cross-shore structures. Although the model skill for mean tracer concentration is variable (from negative to 0.73) depending upon release, cross-shore integrated tracer moments (normalized by the cross-shore tracer integral) have consistently high skills (≈0.9). Modeled and observed bulk surf zone cross-shore diffusivity estimates are also similar, with 0.72 squared correlation and skill of 0.4. Similar to the observations, the model bulk (absolute) cross-shore diffusivity is consistent with a mixing length parameterization based on low-frequency (0.001–0.03 Hz) eddies. The model absolute cross-shore dispersion is dominated by stirring from surf zone eddies and does not depend upon the presence of the breaking wave eddy diffusivity. Given only the bathymetry and incident wave field, the coupled Boussinesq-tracer model qualitatively reproduces the observed cross-shore absolute tracer dispersion, suggesting that the model can be used to study surf zone tracer dispersion mechanisms.This research was supported by SCCOOS, CA Coastal Conservancy, NOAA, NSF, ONR, and CA Sea Grant.2012-05-1

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