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

Dualization of non-Abelian BF model

We show that dualization of BF models to Stueckelberg-like massive gauge theories allows a non-Abelian extension. We obtain local Lagrangians which are straightforward extensions of the Abelian results.Comment: 6 pages, ReVTeX, no figures, to be publ. on Phys.Lett.

The Impact of Pedestrian Crossing Flags on Driver Yielding Behavior in Las Vegas, NV

Walking is the most affordable, accessible, and environmentally friendly method of transportation. However, the risk of pedestrian injury or death from motor vehicle crashes is significant, particularly in sprawling metropolitan areas. The purpose of this study was to examine the effect of pedestrian crossing flags (PCFs) on driver yielding behaviors. Participants crossed a marked, midblock crosswalk on a multilane road in Las Vegas, Nevada, with and without PCFs, to determine if there were differences in driver yielding behaviors (n = 160 crossings). Trained observers recorded (1) the number of vehicles that passed in the nearest lane without yielding while the pedestrian waited at the curb and (2) the number of vehicles that passed through the crosswalk while the pedestrian was in the same half of the roadway. ANOVA revealed that drivers were significantly less likely to pass through the crosswalk with the pedestrian in the roadway when they were carrying a PCF (M = 0.20; M = 0.06); drivers were more likely to yield to the pedestrian waiting to enter the roadway when they were carrying a PCF (M = 1.38; M = 0.95). Pedestrian crossing flags are a low-tech, low-cost intervention that may improve pedestrian safety at marked mid-block crosswalks. Future research should examine driver fade-out effects and more advanced pedestrian safety alternatives

Severe loss-of-function mutations in the adrenocorticotropin receptor (ACTHR, MC2R) can be found in patients diagnosed with salt-losing adrenal hypoplasia

Objective: Familial glucocorticoid deficiency type I (FGD1) is a rare form of primary adrenal insufficiency resulting from recessive mutations in the ACTH receptor (MC2R, MC2R). Individuals with this condition typically present in infancy or childhood with signs and symptoms of cortisol insufficiency, but disturbances in the renin-angiotensin system, aldosterone synthesis or sodium homeostasis are not a well-documented association of FGD1. As ACTH stimulation has been shown to stimulate aldosterone release in normal controls, and other causes of hyponatraemia can occur in children with cortisol deficiency, we investigated whether MC2R changes might be identified in children with primary adrenal failure who were being treated for mineralocorticoid insufficiency. Design: Mutational analysis of MC2R by direct sequencing. Patients: Children (n = 22) who had been diagnosed with salt-losing forms of adrenal hypoplasia (19 isolated cases, 3 familial), and who were negative for mutations in DAX1 (NR0B1) and SF1 (NR5A1). Results: MC2R mutations were found in three individuals or kindred (I: homozygous S74I; II: novel compound heterozygous R146H/560delT; III: novel homozygous 579-581delTGT). These changes represent severely disruptive loss-of-function mutations in this G-protein coupled receptor, including the first reported homozygous frameshift mutation. The apparent disturbances in sodium homeostasis were mild, manifest at times of stress (e.g. infection, salt-restriction, heat), and likely resolved with time. Conclusions: MC2R mutations should be considered in children who have primary adrenal failure with apparent mild disturbances in renin-sodium homeostasis. These children may have been misdiagnosed as having salt-losing adrenal hypoplasia. Making this diagnosis has important implications for treatment, counselling and long-term prognosi

THE BRIDGE WEB SITE GROWING AND SUSTAINING PARTNERSHIPS BETWEEN OCEAN SCIENCE AND EDUCATION

When physicist Tim Berners-Lee and a team of fellow scientists at the European Center for High Energy Physics (CERN) launched the first-ever Web site in 1989, their goal was to make it easier for scientists to access research documents and scientific data (CERN, 2008). In 1998, Virginia Sea Grant educators at the Virginia Institute of Marine Science (VIMS) had a similar goal: to make ocean science educational resources and current research data more accessible to classroom teachers. The Virginia Sea Grant education team took the first step toward accomplishing this goal by launching a Web site of its own, called "Bridge." The name was inspired by the idea of a ship's bridge with a teacher at the helm, navigating "an ocean of marine education data." It also represents a bridge spanning the divide between the education and the ocean research communities, which is the essence of the Bridge project's mission

Quantizing the electromagnetic field near two-sided semitransparent mirrors

This paper models light scattering through flat surfaces with finite transmission, reflection, and absorption rates, with wave packets approaching the mirror from both sides. While using the same notion of photons as in free space, our model also accounts for the presence of mirror images and the possible exchange of energy between the electromagnetic field and the mirror surface. To test our model, we derive the spontaneous decay rate and the level shift of an atom in front of a semitransparent mirror as a function of its transmission and reflection rates. When considering limiting cases and using standard approximations, our approach reproduces well-known results but it also paves the way for the modeling of more complex scenarios

The effects of verbal information on children's fear beliefs about social situations

Two experiments explored the role of verbal information in changing children’s fearrelated beliefs about social situations. In Experiment 1, 118 6- to 8- and 12- to 13-year-olds heard positive, negative, or no information about individuals’ experiences of three social situations. Fear beliefs regarding each situation were assessed before and after this manipulation. Verbal information had no significant influence on children’s fear beliefs. In Experiment 2, the same paradigm was used with 80 12- to 13-year-olds, but the information took the form of multiple attitude statements about the situations expressed by groups of peers, older children, or adults. An affective priming task of implicit attitudes was used to complement the explicit questions about fear beliefs. Negative information influenced both explicit and implicit fear beliefs. The source of information and the child’s own social anxiety did not moderate these effects. Implications for our understanding of the socialisation of childhood fears are discussed

Orbital parameters of supergiant fast X-ray transients

Supergiant fast X-ray transient (SFXT) is a new class of the high mass X-ray binary that shows short X-ray flares. The physical mechanism of SFXT short flares is still open for discussion. The accretion process of dense clumps in stellar wind onto neutron star (NS) has been proposed as the origin of such short flares. In order to examine the applicability of the clumpy wind scenario, we focus on the accretion mode that depends on orbital parameters. Our goal is to impose restrictions on the orbital parameters of SFXT. Assuming a simple analytic model of clumpy wind, we investigate the condition where the size of accretion cylinder overcomes the clump size. The allowed parameter region for SFXT is restricted in a relatively narrow window in $P_{\rm{orb}} - e$ diagram. Binary systems with large eccentricities (e \ga 0.4) and moderate orbital periods ($P_{\rm{orb}} \sim 10$ d) are prone to show periodic X-ray outbursts which are characteristic for SFXT. We confirm that systems with a long orbital period of more than 100 days cannot produce bright X-ray flares in the simple clumpy wind scenario.Comment: 6 pages, accepted for publication in A&

Mutations on the N-terminal edge of the DELSEED loop in either the α or β subunit of the mitochondrial F 1 -ATPase enhance ATP hydrolysis in the absence of the central γ rotor

F1-ATPase is a rotary molecular machine with a subunit stoichiometry of α3 β3 γ1 δ1 ε1. It has a robust ATP-hydrolyzing activity due to effective cooperativity between the three catalytic sites. It is believed that the central γ rotor dictates the