In vitro validation of phosphorodiamidate morpholino oligomers

One of the crucial aspects of screening antisense oligonucleotides destined for therapeutic application is confidence that the antisense oligomer is delivered efficiently into cultured cells. Efficient delivery is particularly vital for antisense phosphorodiamidate morpholino oligomers, which have a neutral backbone, and are known to show poor gymnotic uptake. Here, we report several methods to deliver these oligomers into cultured cells. Although 4D-NucleofectorTM or Neon. electroporation systems provide efficient delivery and use lower amounts of phosphorodiamidate morpholino oligomer, both systems are costly. We show that some readily available transfection reagents can be used to deliver phosphorodiamidate morpholino oligomers as efficiently as the electroporation systems. Among the transfection reagents tested, we recommend Lipofectamine 3000TM for delivering phosphorodiamidate morpholino oligomers into fibroblasts and Lipofectamine 3000TM or Lipofectamine 2000. for myoblasts/myotubes. We also provide optimal programs for nucleofection into various cell lines using the P3 Primary Cell 4D-NucleofectorTM X Kit (Lonza), as well as antisense oligomers that redirect expression of ubiquitously expressed genes that may be used as positive treatments for human and murine cell transfections

International Journal of Fatigue, Volume 54:Evaluating surface deformation and near surface strain hardening resulting from shot peening a tempered martensitic steel and application to low cycle fatigue

The plastic deformation resulting from shot peening treatments applied to the ferritic heat resistant steelFV448 has been investigated. Two important effects have been quantified: surface roughness and strainhardening. 2D and 3D tactile and optical techniques for determining surface roughness amplitude parametershave been investigated; it was found that whilst Ra and Sa were consistent, Sz was generally higherthan Rz due to the increased probability of finding the worst case surface feature. Three different methodsfor evaluating the plastic strain profile have been evaluated with a view to establishing the variation inyield strength near the surface of a shot peened component. Microhardness, X-ray diffraction (XRD) linebroadening and electron backscatter diffraction (EBSD) local misorientation techniques were applied toboth uniaxially deformed calibration samples of known plastic strain and samples shot peened at intensitiesvarying from 4A to 18A to establish the variation in plastic strain and hence the variation in yieldstrength. The results from the three methods were compared; XRD and EBSD profiles were found to bethe most similar with microhardness profiles extending much deeper into the sample. Changes in themeasured plastic strain profile after exposure to low cycle fatigue and the correlation of these changeswith the cyclic stress–strain behaviour of the material are also discussed with a view to assessing theimportance of the dislocation profile in component life assessment procedures. 2013 Elsevier Ltd. All rights reserved

Longitudinal metabolic and gut bacterial profiling of pregnant women with previous bariatric surgery

Objective Due to the global increase in obesity rates and success of bariatric surgery in weight reduction, an increasing number of women now present pregnant with a previous bariatric procedure. This study investigates the extent of bariatric-associated metabolic and gut microbial alterations during pregnancy and their impact on fetal development. Design A parallel metabonomic (molecular phenotyping based on proton nuclear magnetic resonance spectroscopy) and gut bacterial (16S ribosomal RNA gene amplicon sequencing) profiling approach was used to determine maternal longitudinal phenotypes associated with malabsorptive/mixed (n=25) or restrictive (n=16) procedures, compared with women with similar early pregnancy body mass index but without bariatric surgery (n=70). Metabolic profiles of offspring at birth were also analysed. Results Previous malabsorptive, but not restrictive, procedures induced significant changes in maternal metabolic pathways involving branched-chain and aromatic amino acids with decreased circulation of leucine, isoleucine and isobutyrate, increased excretion of microbial-associated metabolites of protein putrefaction (phenylacetlyglutamine, p-cresol sulfate, indoxyl sulfate and p-hydroxyphenylacetate), and a shift in the gut microbiota. The urinary concentration of phenylacetylglutamine was significantly elevated in malabsorptive patients relative to controls (p=0.001) and was also elevated in urine of neonates born from these mothers (p=0.021). Furthermore, the maternal metabolic changes induced by malabsorptive surgery were associated with reduced maternal insulin resistance and fetal/birth weight. Conclusion Metabolism is altered in pregnant women with a previous malabsorptive bariatric surgery. These alterations may be beneficial for maternal outcomes, but the effect of elevated levels of phenolic and indolic compounds on fetal and infant health should be investigated further

Cystic Fibrosis Foundation and European Cystic Fibrosis Society Survey of cystic fibrosis mental health care delivery

Background: Psychological morbidity in individuals with cystic fibrosis (CF) and their caregivers is common. The Cystic Fibrosis Foundation (CFF) and European Cystic Fibrosis Society (ECFS) Guidelines Committee on Mental Health sought the views of CF health care professionals concerning mental health care delivery. Methods: An online survey which focused on the current provision and barriers to mental health care was distributed to CF health care professionals. Results: Of the 1454 respondents, many did not have a colleague trained in mental health issues and 20% had no one on their team whose primary role was focused on assessing or treating these issues. Insufficient resources and a lack of competency were reported in relation to mental health referrals. Seventy-three percent of respondents had no experience with mental health screening. Of those who did, they utilized 48 different, validated scales. Conclusions: These data have informed the decision-making, dissemination and implementation strategies of the Mental Health Guidelines Committee sponsored by the CFF and ECFS

Attachment Styles Within the Coach-Athlete Dyad: Preliminary Investigation and Assessment Development

The present preliminary study aimed to develop and examine the psychometric properties of a new sport-specific self-report instrument designed to assess athletes’ and coaches’ attachment styles. The development and initial validation comprised three main phases. In Phase 1, a pool of items was generated based on pre-existing self-report attachment instruments, modified to reflect a coach and an athlete’s style of attachment. In Phase 2, the content validity of the items was assessed by a panel of experts. A final scale was developed and administered to 405 coaches and 298 athletes (N = 703 participants). In Phase 3, confirmatory factor analysis of the obtained data was conducted to determine the final items of the Coach-Athlete Attachment Scale (CAAS). Confirmatory factor analysis revealed acceptable goodness of fit indexes for a 3-first order factor model as well as a 2-first order factor model for both the athlete and the coach data, respectively. A secure attachment style positively predicted relationship satisfaction, while an insecure attachment style was a negative predictor of relationship satisfaction. The CAAS revealed initial psychometric properties of content, factorial, and predictive validity, as well as reliability

Généralisation du diagramme de Voronoï et placement de formes géométriques complexes dans un nuage de points.

La géométrie algorithmique est une discipline en pleine expansion dont l'objet est la conception d'algorithmes résolvant des problèmes géométriques. De tels algorithmes sont très utiles notamment dans l'ingénierie, l'industrie et le multimédia. Pour être performant, il est fréquent qu'un algorithme géométrique utilise des structures de données spécialisées.Nous nous sommes intéressés à une telle structure : le diagramme de Voronoï et avons proposé une généralisation de celui-ci. Ladite généralisation résulte d'une extension du prédicat du disque vide (prédicat propre à toute région de Voronoï) à une union de disques. Nous avons analysé les régions basées sur le prédicat étendu et avons proposé des méthodes pour les calculer par ordinateur.Par ailleurs, nous nous sommes intéressés aux problèmes de placement de formes , thème récurrent en géométrie algorithmique. Nous avons introduit un formalisme universel pour de tels problèmes et avons, pour la première fois, proposé une méthode de résolution générique, en ce sens qu'elle est apte à résoudre divers problèmes de placement suivant un même algorithme.Nos travaux présentent, d'une part, l'avantage d'élargir le champ d'application de structures de données basées sur Voronoï. D'autre part, ils facilitent de manière générale l'utilisation de la géométrie algorithmique, en unifiant définitions et algorithmes associés aux problèmes de placement de formes.Computational geometry is an active branch of computer science whose goal is the design of efficient algorithms solving geometric problems. Such algorithms are useful in domains like engineering, industry and multimedia. In order to be efficient, algorithms often use special data structures.In this thesis we focused on such a structure: the Voronoi diagram. We proposed a new generalized diagram. We have proceeded by extending the empty disk predicate (satisfied by every Voronoi region) to an arbitrary union of disks. We have analyzed the new plane regions based on the extended predicate, and we designed algorithms for computing them.Then, we have considered another topic, which is related to the first one: shape placement problems. Such problems have been studied repeatedly by researchers in computational geometry. We introduced new notations along with a global framework for such problems. We proposed, for the first time a generic method, which is able to solve various placement problems using a single algorithm.Thus, our work extend the scope of Voronoi based data structures. It also simplifies the practical usage of placement techniques by unifying the associated definitions and algorithms.MULHOUSE-SCD Sciences (682242102) / SudocSudocFranceF

Predictors of lane-change errors in older drivers

OBJECTIVES: To determine the factors that predict errors in executing proper lane changes among older drivers. DESIGN: Cross-sectional analysis of data from a longitudinal study. SETTING: Maryland's Eastern Shore. PARTICIPANTS: One thousand eighty drivers aged 67 to 87 enrolled in the Salisbury Eye Evaluation Driving Study. MEASUREMENTS: Tests of vision, cognition, health status, and self-reported distress and a driving monitoring system in each participant's car, used to quantify lane-change errors. RESULTS: In regression models, measures of neither vision nor perceived stress were related to lane-change errors after controlling for age, sex, race, and residence location. In contrast, cognitive variables, specifically performance on the Brief Test of Attention and the Beery-Buktenicka Test of Visual-Motor Integration, were related to lane-change errors. CONCLUSION: The current findings underscore the importance of specific cognitive skills, particularly auditory attention and visual perception, in the execution of driving maneuvers in older individuals

Sex differences in cognition in healthy elderly individuals

Sex differences in patterns of cognitive test performance have been attributed to factors, such as sex hormones or sexual dimorphisms in brain structure, that change with normal aging. The current study examined sex differences in patterns of cognitive test performance in healthy elderly individuals. Cognitive test scores of 957 men and women (age 67-89), matched for overall level of cognitive test performance, age, education, and depression scale score, were compared. Men and women were indistinguishable on tests of auditory divided attention, category fluency, and executive functioning. In contrast, women performed better than men on tests of psychomotor speed and verbal learning and memory, whereas men outperformed women on tests of visuoconstruction and visual perception. Our finding that the pattern of sex differences in cognition observed in young adults is observed in old age has implications for future studies of both healthy elderly individuals and of those with cognitive disorders

Visual and cognitive deficits predict stopping or restricting driving: The salisbury eye evaluation driving study (SEEDS)

PURPOSE. To determine the visual and other factors that predict stopping or restricting driving in older drivers. METHODS. A group of 1425 licensed drivers aged 67 to 87 years, who were residents of greater Salisbury, participated. At 1 year after enrollment, this group was categorized into those who had stopped driving, drove only within their neighborhood, or continued to drive beyond their neighborhood. At baseline, a battery of structured questionnaires, vision, and cognitive tests were administered. Multivariate analysis determined the factors predictive of stopping or restricting driving 12 months later. RESULTS. Of the 1425 enrolled, 1237 (87%) were followed up at 1 year. Excluding those who were already limiting their driving at baseline (n = 35), 1.5% (18/1202) had stopped and 3.4% (41/1202) had restricted their driving. The women (odds ratio [OR], 4.01; 95% confidence interval [CI], 2.05- 8.20) and those who prefer to be driven (OR, 3.91; 95% CI, 1.91- 8.00) were more likely to stop or restrict driving. Depressive symptoms increased likelihood of restricting or stopping driving (OR, 1.08; 95% CI, 1.009 -1.16 per point Geriatric Depression Scale). Slow visual scanning and psychomotor speed (Trail Making Test, Part A: OR, 1.02; 95% CI, 1.01-1.03), poor visuoconstructional skills (Beery-Buktenica Test of Visual Motor Integration: OR, 1.14; 95% CI, 1.05-1.25), and reduced contrast sensitivity (OR, 1.15; 95% CI, 1.03-1.28) predicted stopping or reducing driving. Visual field loss and visual attention were not associated. The effect of vision on changing driving behavior was partially mediated by cognition, depression, and baseline driving preferences. CONCLUSIONS. In this cohort, contrast sensitivity and cognitive function were independently associated with incident cessation or restriction of driving space. These data suggest drivers with functional deficits make difficult decisions to restrict or stop driving

“Strong Teeth”: the acceptability of an early-phase feasibility trial of an oral health intervention delivered by dental teams to parents of young children

Background Dental caries (tooth decay) in children is a worldwide public health problem. The leading cause of caries is poor oral hygiene behaviours and the frequent consumption of sugary foods and drinks. Changing oral health habits requires effective behaviour change conversations. The dental practice provides an opportunity for dental teams to explore with parents the oral health behaviours they undertake for their young children (0–5 years old). However, evidence suggests that dental teams need further support, training and resources. Therefore, “Strong Teeth” (an oral health intervention) was co-developed to help dental teams undertake these behaviour change conversations. The current paper will explore the acceptability of the “Strong Teeth” intervention with dental teams and parents of children aged 0–5 years old using multiple datasets (interviews, focus groups and dental team member diaries) Methods Following the delivery of the “Strong Teeth” intervention, qualitative interviews with parents and focus groups with dental team members were undertaken. Interviews were audio-recorded, transcribed and analysed using a theoretical framework of acceptability. The self-reported dental team diaries supplemented the interviews and focus groups and were analysed using framework analysis. Results Four themes were developed: (1) integration within the dental practice; (2) incorporating the Oral-B electric toothbrush; (3) facilitating discussions and demonstrations; and (4) the practicality of the Disney Magic Timer app. Overall, the “Strong Teeth” intervention was acceptable to parents and dental teams. Parents felt the Oral-B electric toothbrush was a good motivator; however, the Disney Magic Timer app received mixed feedback on how well it could be used effectively in the home setting. Findings suggest that the intervention was more acceptable as a “whole team approach” when all members of the dental practice willingly participated. Conclusions There are limited studies that use a robust process evaluation to measure the acceptability of an intervention. The use of the theoretical framework of acceptability helped identify aspects of the intervention that were positive and helped identify the interventions areas for enhancement moving forwards. Future modifications include enhanced whole team approach training to optimise acceptability to all those involved. Trial registration ISRCTN Register, (ISRCTN10709150)