MesoTIRF: a novel axial super-resolution illuminator for membrane imaging over a 4.4 mm x 3.0 mm field of view

In Total Internal Reflection Fluorescence (TIRF) microscopy, a specimen is illuminated by the evanescent field produced by a beam undergoing total internal reflection, whose characteristic depth is orders of magnitude below the axial diffraction limit. The axial super-resolution and much improved contrast of TIRF gives a substantially improved signal-to-background ratio (SBR) than that which can be achieved with widefield illumination, and it is used extensively for imaging of the cell membrane [1]– [3] . Commercial TIRF objectives allow for a simple adaption to existing microscope systems. To attain a super-critical angle at the specimen plane, the illumination must enter the back focal plane of these objectives off-axis, requiring a high numerical aperture. As such, the magnification of these objectives is generally a minimum of 60x, reducing the lateral imaging field to less than 100 µm in diameter. Sub-cellular axial resolution is therefore restricted to a tiny population of cells and statistically significant data sampling may be difficult to achieve. To address this, we have developed a TIRF illuminator for the Mesolens, a custom giant objective lens with a 4x/0.47NA specification. The Mesolens provides an imaging field of 4.4 mm x 3.0 mm, and in combination with our new TIRF illuminator which we call MesoTIRF we have performed imaging of large cell populations with sub-micron resolution in three-dimensions. With MesoTIRF we demonstrate more than a 5-fold improvement in SBR and significantly reduced photobleaching rate compared to widefield epifluorescence illumination. We will present details of the MesoTIRF system together with current and emerging applications in cell imaging

Prevalence and Predictors of Vitamin D Insufficiency in Children: A Great Britain Population Based Study

Objectives To evaluate the prevalence and predictors of vitamin D insufficiency (VDI) in children In Great Britain. Design A nationally representative cross-sectional study survey of children (1102) aged 4–18 years (999 white, 570 male) living in private households (January 1997–1998). Interventions provided information about dietary habits, physical activity, socio-demographics, and blood sample. Outcome measures were vitamin D insufficiency (<50 nmol/L). Results Vitamin D levels (mean = 62.1 nmol/L, 95%CI 60.4–63.7) were insufficient in 35%, and decreased with age in both sexes (p<0.001). Young People living between 53–59 degrees latitude had lower levels (compared with 50–53 degrees, p = 0.045). Dietary intake and gender had no effect on vitamin D status. A logistic regression model showed increased risk of VDI in the following: adolescents (14–18 years old), odds ratio (OR) = 3.6 (95%CI 1.8–7.2) compared with younger children (4–8 years); non white children (OR = 37 [95%CI 15–90]); blood levels taken December-May (OR = 6.5 [95%CI 4.3–10.1]); on income support (OR = 2.2 [95%CI 1.3–3.9]); not taking vitamin D supplementation (OR = 3.7 [95%CI 1.4–9.8]); being overweight (OR 1.6 [95%CI 1.0–2.5]); <1/2 hour outdoor exercise/day/week (OR = 1.5 [95%CI 1.0–2.3]); watched >2.5 hours of TV/day/week (OR = 1.6[95%CI 1.0–2.4]). Conclusion We confirm a previously under-recognised risk of VDI in adolescents. The marked higher risk for VDI in non-white children suggests they should be targeted in any preventative strategies. The association of higher risk of VDI among children who exercised less outdoors, watched more TV and were overweight highlights potentially modifiable risk factors. Clearer guidelines and an increased awareness especially in adolescents are needed, as there are no recommendations for vitamin D supplementation in older children

On opportunistic software reuse

The availability of open source assets for almost all imaginable domains has led the software industry toopportunistic design-an approach in which people develop new software systems in an ad hoc fashion by reusing and combining components that were not designed to be used together. In this paper we investigate this emerging approach. We demonstrate the approach with an industrial example in whichNode.jsmodules and various subsystems are used in an opportunistic way. Furthermore, to study opportunistic reuse as a phenomenon, we present the results of three contextual interviews and a survey with reuse practitioners to understand to what extent opportunistic reuse offers improvements over traditional systematic reuse approaches.Peer reviewe

Are GPs adequately equipped with the knowledge for educating and counseling of families with ADHD children?

<p>Abstract</p> <p>Background</p> <p>Attention deficit hyperactivity disorder is one of the most common child psychiatry disorders. General physicians (GP), as primary care providers, can have an important role in screening and treatment of ADHD. This study aimed to survey GPs' knowledge, attitude, and their views of their role in the screening, diagnosing and managing children with ADHD.</p> <p>Methods</p> <p>Six hundred and sixty five general physicians in Shiraz, Iran, answered a self-reported questionnaire on ADHD. The questionnaire consisted of questions regarding socio-demographic characteristics such as age, the duration of practice as a GP, marital status, general knowledge about ADHD, and the management of ADHD.</p> <p>Results</p> <p>Less than half of them believed that they have adequate knowledge and information about this disorder. They usually do not like to be the primary care providers for children with ADHD. The majority of them prefer to refer the children to related specialists, mostly psychiatrists or psychologists. More than one third of them believed that sugar is a cause of ADHD. Only 6.6% of them reported that ADHD persists for the whole life. Their knowledge about methylphenidate is reasonable.</p> <p>Conclusions</p> <p>As many other countries worldwide, the knowledge of GPs about ADHD should be improved. They do not asses and manage children with probable ADHD by themselves without referring to related professionals. They do not opt for the use of methylphenidate.</p

Morpholino Gene Knockdown in Adult Fundulus heteroclitus: Role of SGK1 in Seawater Acclimation

The Atlantic killifish (Fundulus heteroclitus) is an environmental sentinel organism used extensively for studies on environmental toxicants and salt (NaCl) homeostasis. Previous research in our laboratory has shown that rapid acclimation of killifish to seawater is mediated by trafficking of CFTR chloride channels from intracellular vesicles to the plasma membrane in the opercular membrane within the first hour in seawater, which enhances chloride secretion into seawater, thereby contributing to salt homeostasis. Acute transition to seawater is also marked by an increase in both mRNA and protein levels of serum glucocorticoid kinase 1 (SGK1) within 15 minutes of transfer. Although the rise in SGK1 in gill and its functional analog, the opercular membrane, after seawater transfer precedes the increase in membrane CFTR, a direct role of SGK1 in elevating membrane CFTR has not been established in vivo. To test the hypothesis that SGK1 mediates the increase in plasma membrane CFTR we designed two functionally different vivo-morpholinos to knock down SGK1 in gill, and developed and validated a vivo-morpholino knock down technique for adult killifish. Injection (intraperitoneal, IP) of the splice blocking SGK1 vivo-morpholino reduced SGK1 mRNA in the gill after transition from fresh to seawater by 66%. The IP injection of the translational blocking and splice blocking vivo-morpholinos reduced gill SGK1 protein abundance in fish transferred from fresh to seawater by 64% and 53%, respectively. Moreover, knock down of SGK1 completely eliminated the seawater induced rise in plasma membrane CFTR, demonstrating that the increase in SGK1 protein is required for the trafficking of CFTR from intracellular vesicles in mitochondrion rich cells to the plasma membrane in the gill during acclimation to seawater. This is the first report of the use of vivo-morpholinos in adult killifish and demonstrates that vivo-morpholinos are a valuable genetic tool for this environmentally relevant model organism

The role of multi-slice computed tomography in stable angina management: a current perspective

Contrast-enhanced CT coronary angiography (CTCA) has evolved as a reliable alternative imaging modality technique and may be the preferred initial diagnostic test in patients with stable angina with intermediate pre-test probability of CAD. However, because CTCA is moderately predictive for indicating the functional significance of a lesion, the combination of anatomic and functional imaging will become increasingly important. The technology will continue to improve with better spatial and temporal resolution at low radiation exposure, and CTCA may eventually replace invasive coronary angiography. The establishment of the precise role of CTCA in the diagnosis and management of patients with stable angina requires high-quality randomised study designs with clinical outcomes as a primary outcome

Detection of Gamma-Ray Emission from the Starburst Galaxies M82 and NGC 253 with the Large Area Telescope on Fermi

We report the detection of high-energy gamma-ray emission from two starburst galaxies using data obtained with the Large Area Telescope on board the Fermi Gamma-ray Space Telescope. Steady point-like emission above 200 MeV has been detected at significance levels of 6.8 sigma and 4.8 sigma respectively, from sources positionally coincident with locations of the starburst galaxies M82 and NGC 253. The total fluxes of the sources are consistent with gamma-ray emission originating from the interaction of cosmic rays with local interstellar gas and radiation fields and constitute evidence for a link between massive star formation and gamma-ray emission in star-forming galaxies.Comment: Submitted to ApJ Letter

Incremental value of the CT coronary calcium score for the prediction of coronary artery disease

Objectives:: To validate published prediction models for the presence of obstructive coronary artery disease (CAD) in patients with new onset stable typical or atypical angina pectoris and to assess the incremental value of the CT coronary calcium score (CTCS). Methods:: We searched the literature for clinical prediction rules for the diagnosis of obstructive CAD, defined as≥50% stenosis in at least one vessel on conventional coronary angiography. Significant variables were re-analysed in our dataset of 254 patients with logistic regression. CTCS was subsequently included in the models. The area under the receiver operating characteristic curve (AUC) was calculated to assess diagnostic performance. Results:: Re-analysing the variables used by Diamond & Forrester yielded an AUC of 0.798, which increased to 0.890 by adding CTCS. For Pryor, Morise 1994, Morise 1997 and Shaw the AUC increased from 0.838 to 0.901, 0.831 to 0.899, 0.840 to 0.898 and 0.833 to 0.899. CTCS significantly improved model performance in each model. Conclusions:: Validation demonstrated good diagnostic performance across all models. CTCS improves the prediction of the presence of obstructive CAD, independent of clinical predictors, and should be considered in its diagnostic work-up. © 2010 The Author(s)