Effect of delayed acquisition times on Gadolinium-enhanced MRI of the presumably normal canine brain

A delay in imaging following intravenous contrast medium administration has been recommended to reduce misdiagnoses. However, the normal variation of contrast enhancement in dogs following a delay has not been characterized. Contrast enhanced MR imaging of 22 dogs was assessed, in terms of identification of normal anatomic structures, to investigate the variation associated with 10 minute delay between contrast medium administration and imaging. All dogs had a normal brain MR imaging study and unremarkable CSF. Specific ROIs were assessed both objectively, using computer software, and subjectively using three observers. Mean contrast enhancement greater than 10% was seen in the pituitary gland, choroid plexus, meninges, temporal muscle, trigeminal nerve and the trigeminal nerve root. Structures with an active blood-brain-barrier had minimal contrast enhancement (<6%). Enhancing structures had significantly more contrast enhancement at t=1min versus t=10min, except in temporal muscle, the trigeminal nerve and the trigeminal nerve root. Inter-observer agreement was moderate to good in favor of the initial post contrast T1w sequence. The observers found either no difference or poor agreement in identification of the non-vascular structures. Intra-observer agreement was very good with all vascular structures and most non-vascular structures. A degree of meningeal enhancement was a consistent finding. The initial acquisition had higher enhancement characteristics and observer agreement for some structures; however, contrast-to-noise was comparable in the delayed phase or not significantly different. We provide baseline references and suggest that the initial T1w post contrast sequence is preferable but not essential should a delayed post contrast T1w sequence be performed

Revalidation and electronic cataract surgery audit: a Scottish survey on current practice and opinion

PURPOSE: To determine current knowledge and opinion on revalidation, and methods of cataract surgery audit in Scotland and to outline the current and future possibilities for electronic cataract surgery audit. METHODS: In 2010 we conducted a prospective, cross-sectional, Scottish-wide survey on revalidation knowledge and opinion, and cataract audit practice among all senior NHS ophthalmologists. Results were anonymised and recorded manually for analysis. RESULTS: In all, 61% of the ophthalmologists surveyed took part. Only 33% felt ready to take part in revalidation, whereas 76% felt they did not have adequate information about the process. Also, 71% did not feel revalidation would improve patient care, but 85% agreed that cataract surgery audit is essential for ophthalmic practice. In addition, 91% audit their cataract outcomes; 52% do so continuously. Further, 63% audit their subspecialist surgical results. Only 25% audit their cataract surgery practice electronically, and only 12% collect clinical data using a hospital PAS system. Funding and system incompatibility were the main reasons cited for the lack of electronic audit setup. Currently, eight separate hospital IT patient administration systems are used across 14 health boards in Scotland. CONCLUSION: Revalidation is set to commence in 2012. The Royal College of Ophthalmologists will use cataract outcome audit as a tool to ensure surgical competency for the process. Retrospective manual auditing of cataract outcome is time consuming, and can be avoided with an electronic system. Scottish ophthalmologists view revalidation with scepticism and appear to have inadequate knowledge of the process. However, they strongly agree with the concept of cataract surgery audit. The existing and future electronic applications that may support surgical audit are commercial electronic records, web-based applications, centrally funded software applications, and robust NHS connections between community and hospital

Three-dimensional conformation at the H19/Igf2 locus supports a model of enhancer tracking

Insight into how the mammalian genome is structured in vivo is key to understanding transcriptional regulation. This is especially true in complex domains in which genes are coordinately regulated by long-range interactions between cis-regulatory elements. The regulation of the H19/Igf2 imprinted region depends on the presence of several cis-acting sequences, including a methylation-sensitive insulator between Igf2 and H19 and shared enhancers downstream of H19. Each parental allele has a distinct expression pattern. We used chromosome conformation capture to assay the native three-dimensional organization of the H19/Igf2 locus on each parental copy. Furthermore, we compared wild-type chromosomes to several mutations that affect the insulator. Our results show that promoters and enhancers reproducibly co-localize at transcriptionally active genes, i.e. the endodermal enhancers contact the maternal H19 and the paternal Igf2 genes. The active insulator blocks traffic of the enhancers along the chromosome, restricting them to the H19 promoter. Conversely, the methylated inactive insulator allows the enhancers to contact the upstream regions, including Igf2. Mutations that either remove or inhibit insulator activity allow unrestricted access of the enhancers to the whole region. A mutation that allows establishment of an enhancer-blocker on the normally inactive paternal copy diminishes the contact of the enhancer with the Igf2 gene. Based on our results, we propose that physical proximity of cis-acting DNA elements is vital for their activity in vivo. We suggest that enhancers track along the chromosome until they find a suitable promoter sequence to interact with and that insulator elements block further tracking of enhancers

Closed Universes, de Sitter Space and Inflation

We present a new approach to constructing inflationary models in closed universes. Conformal embedding of closed-universe models in a de Sitter background suggests a quantisation condition on the available conformal time. This condition implies that the universe is closed at no greater than the 10% level. When a massive scalar field is introduced to drive an inflationary phase this figure is reduced to closure at nearer the 1% level. In order to enforce the constraint on the available conformal time we need to consider conditions in the universe before the onset of inflation. A formal series around the initial singularity is constructed, which rests on a pair of dimensionless, scale-invariant parameters. For physically-acceptable models we find that both parameters are of order unity, so no fine tuning is required, except in the mass of the scalar field. For typical values of the input parameters we predict the observed values of the cosmological parameters, including the magnitude of the cosmological constant. The model produces a very good fit to the most recent CMBR data. The primordial curvature spectrum predicts the low-l fall-off in the CMB power spectrum observed by WMAP. The spectrum also predicts a fall-off in the matter spectrum at high k, relative to a power law. A further prediction of our model is a large tensor mode component, with r~0.2.Comment: 38 pages, 25 figure

Prostaglandin E2 Prevents Hyperosmolar-Induced Human Mast Cell Activation through Prostanoid Receptors EP2 and EP4

Background: Mast cells play a critical role in allergic and inflammatory diseases, including exercise-induced bronchoconstriction (EIB) in asthma. The mechanism underlying EIB is probably related to increased airway fluid osmolarity that activates mast cells to the release inflammatory mediators. These mediators then act on bronchial smooth muscle to cause bronchoconstriction. In parallel, protective substances such as prostaglandin E2 (PGE2) are probably also released and could explain the refractory period observed in patients with EIB. Objective: This study aimed to evaluate the protective effect of PGE2 on osmotically activated mast cells, as a model of exercise-induced bronchoconstriction. Methods: We used LAD2, HMC-1, CD34-positive, and human lung mast cell lines. Cells underwent a mannitol challenge, and the effects of PGE2 and prostanoid receptor (EP) antagonists for EP1-4 were assayed on the activated mast cells. Beta-hexosaminidase release, protein phosphorylation, and calcium mobilization were assessed. Results: Mannitol both induced mast cell degranulation and activated phosphatidyl inositide 3-kinase and mitogen-activated protein kinase (MAPK) pathways, thereby causing de novo eicosanoid and cytokine synthesis. The addition of PGE2 significantly reduced mannitol-induced degranulation through EP2 and EP4 receptors, as measured by beta-hexosaminidase release, and consequently calcium influx. Extracellular-signal-regulated kinase 1/2, c-Jun N-terminal kinase, and p38 phosphorylation were diminished when compared with mannitol activation alone. Conclusions:Our data show a protective role for the PGE2 receptors EP2 and EP4 following osmotic changes, through the reduction of human mast cell activity caused by calcium influx impairment and MAP kinase inhibition

Eucapnic Voluntary Hyperpnea: Gold Standard for Diagnosing Exercise-Induced Bronchoconstriction in Athletes?

In athletes, a secure diagnos is of exercise-induced bronchoconstriction (EIB) is dependent on objective testing. Evaluating spirometric indices of airflow before and following an exercise bout is intuitively the optimal means for the diagnosis; however, this approach is recognized as having several key limitations. Accordingly, alternative indirect bronchoprovocation tests have been recommended as surrogate means for obtaining a diagnosis of EIB. Of these tests, it is often argued that the eucapnic voluntary hyperpnea (EVH) challenge represents the ‘gold standard’. This article provides a state-of-the-art review of EVH, including an overview of the test methodology and its interpretation. We also address the performance of EVH against the other functional and clinical approaches commonly adopted for the diagnosis of EIB. The published evidence supports a key role for EVH in the diagnostic algorithm for EIB testing in athletes. However, its wide sensitivity and specificity and poor repeatability preclude EVH from being termed a ‘gold standard’ test for EIB

The neurofibromatosis type I pre-mRNA is a novel target of CELF protein-mediated splicing regulation

The CUG-BP and ETR-3 like factors (CELF) are a family of six highly conserved RNA-binding proteins that preferentially bind to UG-rich sequences. One of the key functions of these proteins is to mediate alternative splicing in a number of tissues, including brain, heart and muscle. To fully understand the function of CELF proteins, it is important to identify downstream targets of CELF proteins. In this communication, we report that neurofibromatosis type I (NF1) exon 23a is a novel target of CELF protein-mediated splicing regulation in neuron-like cells. NF1 regulates Ras signaling, and the isoform that excludes exon 23a shows 10 times greater ability to down-regulate Ras signaling than the isoform that includes exon 23a. Five of the six CELF proteins strongly suppress the inclusion of NF1 exon 23a. Over-expression or siRNA knockdown of these proteins in cell transfection experiments altered the levels of NF1 exon 23a inclusion. In vitro binding and splicing analyses demonstrate that CELF proteins block splicing through interfering with binding of U2AF65. These studies, combined with our previous investigations demonstrating a role for Hu proteins and TIA-1/TIAR in controlling NF1 exon 23a inclusion, highlight the complex nature of regulation of this important alternative splicing event