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

‘You’re never too old’: beliefs about physical activity and playing sport in later life

It was evident from interviews with 15 men and women over 70 years of age that the experiences derived from being physically active and playing sport in later life are expansive, dynamic and completely imbued with multiple interpretations. Furthermore, their beliefs support the notion that the physically active body should be considered more than a physical object, as it represents a conscious, feeling, thinking and reflective self. The findings signify much is to be gained by regularly partaking in deliberate physical activity during later life although this is not always as easy as it sounds. At a personal level, changes in functional capacity and dealing with the sometimes ailing body can serve as a deterrent. From a broader perspective, the stigma associated with being older also poses a significant physical, sociological and psychological challenge, particularly during the initial phases. In conclusion, the study discusses why change must occur at a personal and societal level before a greater proportion of the older population become more physically active

Adjustable speed operation of a hydropower plant associated to an irrigation reservoir

This paper deals with the issue of adjustable speed operation (ASO) of hydropower plants. The main idea of this technique is to allow the turbine speed to change in accordance with hydraulic conditions, thus improving the overall unit efficiency. General technical aspects of ASO are further discussed with special emphasis on the energy and operational benefits that may potentially result from its application. In order to assess these benefits, annual operation of a hydropower plant associated to an irrigation reservoir has been simulated under different scenarios, with both adjustable and fixed speed. Turbine operating range proved to be wider with ASO. In addition, simulation results confirm that considerable energy gains are expected to be obtained

The suppression of Curie temperature by Sr doping in diluted ferromagnetic semiconductor (La1-xSrx)(Zn1-yMny)AsO

(La1-xSrx)(Zn1-yMny)AsO is a two dimensional diluted ferromagnetic semiconductor that has the advantage of decoupled charge and spin doping. The substitution of Sr2+ for La3+ and Mn2+ for Zn2+ into the parent semiconductor LaZnAsO introduces hole carriers and spins, respectively. This advantage enables us to investigate the influence of carrier doping on the ferromagnetic ordered state through the control of Sr concentrations in (La1-xSrx)(Zn0.9Mn0.1)AsO. 10 % Sr doping results in a ferromagnetic ordering below TC ~ 30 K. Increasing Sr concentration up to 30 % heavily suppresses the Curie temperature and saturation moments. Neutron scattering measurements indicate that no structural transition occurs for (La0.9Sr0.1)(Zn0.9Mn0.1)AsO below 300 K.Comment: Submitted to EP

Rain Effect Frequency of Infiltration Rate and Infiltration Capacity in Common Soil: Laboratory Test with Rainfall Simulator

Analyzing the Influence of Rain Frequency Infiltration Rate and Infiltration Capacity in Common Soil Type (Laboratory Testing Study With Rainfall Simulator). Infiltration is the flow of water into the ground through the soil surface. This process is a very important part of the hydrological cycle and in the process of transferring rain into the flow of water in the soil before reaching the river. Infiltration (infiltration rate and capacity) is influenced by various variables, including soil type, slope inclination, density and type of vegetation, soil moisture content, and rainfall intensity. This study aims to determine the effect of rainfall frequency on the infiltration rate and infiltration capacity on common soil types. This research is a type of laboratory experimental research, using rainfall simulator tool. The soil used in this study is common soil type. Furthermore, artificial rain was provided with intensity I5, I15, and I25 and performed infiltration rate reading on the Drain Rainfall Simulator. The rate and capacity of infiltration in common soils increase proportionally to the increased intensity of rainfall, the higher the intensity of rainfall the higher the infiltration occurring at the same level of rain frequency. The rate and capacity of infiltration in common soils decrease proportionally to the increasing frequency of rain, the more the frequency of rain the smaller the infiltration occurring at the same level of rainfall intensit

“Bendy” stents help negotiate hairpin intracardiac curves

Simple transposition of the great arteries (TGA) occurs in 0.2 per 1000 live births. The condition is surgically repaired in the neonatal period by the arterial switch procedure (ASO) sometimes preceded by an atrial septostomy. The ASO involves transecting the great arteries and relocating them to the appropriate ventriculo-arterial (VA) connection with attachment of the disconnected coronary arteries to the aorta. In the process, the attachment of the pulmonary artery to the right ventricle involves the Le Compte manoeuvre and to achieve this the pulmonary arteries must be fully mobilised and sometimes the main pulmonary artery may require patch augmentation as well. Nevertheless, pulmonary artery stenosis (PAS) is one of the potential problems with the ASO. However, with improved surgical techniques, this has dropped from around 15% in the 1980s to less than 3%. Apart from surgical revision when PAS occurs, there are interventional options which include angioplasty and/or stent insertion. The latter is preferred in small children and works well in around 60% but may require repeat procedures. In older patients or when angioplasty fails, stent insertion can be considered. These procedures may involve negotiating tight bends in order to reach the site of stenosis. The passage of non-premounted stents may be problematic in such situations, especially with longer stents and tighter bends as they tend to slip off balloon. We describe several techniques that may facilitate such interventions, and these were utilised in an adolescent patient who had had ASO for TGA in the neonatal period. These included manually giving the mounted stent a slight bend in order to help the balloon-stent assembly negotiate hairpin bends.peer-reviewe

Effect of Aso limonite on anaerobic digestion of waste sewage sludge

The effect of Aso volcanic limonite was explored in anaerobic digestion using waste sewage sludge (WSS). In this study, methane and hydrogen sulfide were remarkably inhibited when Aso limonite was mixed with WSS as well as a significant reduction of ammonia. Although pH was lowered after adding Aso limonite, methane was still inhibited in neutralized pH condition at 7.0. Hydrolysis stage was not influenced by Aso limonite as supported by the result that a high protease activity was still detected in the presence of the material. However, acidogenesis stage was affected by Aso limonite as indicated by the different productions of organic acids. Acetic acid, was accumulated in the presence of Aso limonite due to the inhibition of methane production, except in the highest concentration of Aso limonite which the production of acetate may be inhibited. Besides, the production of propionate and butyrate reduced in accordance to the increased concentration of Aso limonite. In addition, Archaeal activity (methanogens) in WSS with Aso limonite was low in agreement with the low methane production. Thus, these results indicate that Aso limonite influences the acidogenesis and methanogenesis processes, by which the productions of methane and ammonia were inhibited. On the other hand, in the contactless of Aso limonite during the anaerobic digestion of WSS (Aso limonite was placed in the area of headspace in the vial), Aso limonite had the adsorptive ability for hydrogen sulfide from WSS, but not for methane. This contactless system of Aso limonite may be a practical means to remove hydrogen sulfide without inhibiting methane production as an important bioenergy source

Why Use the Services of Alternative Staffing Organizations: Perspectives from Customer Businesses

Organizations that aim to improve the experiences and employment chances of job seekers who face barriers to employment have, over the years, had to contend directly with potential employers and their requirements. This is particularly true for community-based job brokers that use a temporary staffing model, offering job access and immediate work to their service population.Alternative staffing organizations (ASOs) are worker-centered, social purpose businesses that place job seekers in temporary and "temp-to-perm" assignments with customer businesses, and charge their customers a markup on the wage of the position. These fee-for-service organizations can help job seekers who face labor market barriers gain work experience and access potential employers. Created by community-based organizations and national nonprofits, ASOs are often embedded within larger organizations that provide other employment, training, and human services to their community. The parent organizations may also be operating other social enterprise ventures. Businesses that contract ASOs for staffing services are customers that expect a service, but also represent an opportunity for employment and work experience for job seekers. Thus ASOs must operate with a dual agenda to serve both sides of the equation. In related publications, we have explored how ASOs operate as social enterprises and how the model fits within the goals of the parent organization. With detailed information from five well-established ASOs, and as part of two waves of a demonstration initiated by the Charles Stewart Mott Foundation, we have documented the employment experiences of workers placed in assignments and their employment status after leaving the ASO. In this paper, we address engagement with businesses and their perspectives on ASO services. This is a major issue for ASOs as well as for other workforce development organizations. ASOs engage with businesses while selling staffing services and monitoring worker performance. By the very nature of temporary staffing, they receive rapid feedback on worker performance and their services from customer businesses. As such, ASOs provide a window into how to connect to potential employers in order to access opportunities. Also, activities of ASOs shed light on how hiring takes place for entry-level jobs, and how customer businesses use ASOs to solve their entry-level hiring problems.This paper demonstrates what can be learned from customers of established ASOs about their reasons for using these services. Specifically, it explores how customer businesses use temporary staffing by ASOs, and for what purposes. What business needs do they meet with ASO services? What are their reasons for using an ASO over conventional staffing agencies? And finally, what causes customer businesses to use an ASO and retain the service over time?These concerns are salient for those organizations considering the creation of an ASO. They also are important for workforce development programs that need to become more active in engaging potential employers and that seek solutions for job seekers who need to connect to employment and need immediate income

Finding the Right Fit: How Alternative Staffing Affects Worker Outcomes

Evaluates implementation and activities at four worker-centered, social-purpose alternative staffing organizations, including worker profiles, jobs secured, experience with the ASO, earnings, and subsequent job status, and business clients' experience