Mutational analysis of a heterogeneous nuclear ribonucleoprotein A2 response element for RNA trafficking

Cytoplasmic transport and localization of mRNA has been reported for a range of oocytes and somatic cells. The heterogeneous nuclear ribonucleoprotein (hnRNP) A2 response element (A2RE) is a 21-nucleotide segment of the myelin basic protein mRNA that is necessary and sufficient for cytoplasmic transport of this message in oligodendrocytes, The predominant A2RE-binding protein in rat brain has previously been identified as hnRNP A2, Here we report that an 11-nucleotide subsegment of the A2RE (A2RE11) was as effective as the full-length A2RE in binding hnRNP A2 and mediating transport of heterologous RNA in oligodendrocytes, Point mutations of the A2RE11 that eliminated binding to hnRNP A2 also markedly reduced the ability of these oligoribonucleotides to support RNA transport, Oligodendrocytes treated with antisense oligonucleotides directed against the translation start site of hnRNP A2 had reduced levels of this protein and disrupted transport of microinjected myelin basic protein RNA. Several A2RE-like sequences from localized neuronal RNAs also bound hnRNP A2 and promoted RNA transport in oligo-dendrocytes, These data demonstrate the specificity of A2RE recognition by hnRNP A2, provide direct evidence for the involvement of hnRNP A2 in cytoplasmic RNA transport, and suggest that this protein may interact with a wide variety of localized messages that possess A2RE-like sequences

The PKC, HOG and Ca2+ signalling pathways co-ordinately regulate chitin synthesis in Candida albicans

Open Access via PMC2649417Peer reviewedPublisher PD

Co-expression of C9orf72 related dipeptide-repeats over 1000 repeat units reveals age-A nd combination-specific phenotypic profiles in Drosophila

A large intronic hexanucleotide repeat expansion (GGGGCC) within the C9orf72 (C9orf72-SMCR8 Complex Subunit) locus is the most prevalent genetic cause of both Frontotemporal Dementia (FTD) and Motor Neuron Disease (MND). In patients this expansion is typically hundreds to thousands of repeat units in length. Repeat associated non-AUG translation of the expansion leads to the formation of toxic, pathological Dipeptide-Repeat Proteins (DPRs). To date there remains a lack of in vivo models expressing C9orf72 related DPRs with a repeat length of more than a few hundred repeats. As such our understanding of how physiologically relevant repeat length DPRs effect the nervous system in an ageing in vivo system remains limited. In this study we generated Drosophila models expressing DPRs over 1000 repeat units in length, a known pathological length in humans. Using these models, we demonstrate each DPR exhibits a unique, age-dependent, phenotypic and pathological profile. Furthermore, we show co-expression of specific DPR combinations leads to distinct, age-dependent, phenotypes not observed through expression of single DPRs. We propose these models represent a unique, in vivo, tool for dissecting the molecular mechanisms implicated in disease pathology, opening up new avenues in the study of both MND and FTD

What influences speech-language pathologists' use of different types of language assessments for elementary school-age children?

Purpose: This study reports on data from a survey of speech-language pathologists' (SLPs) language assessment practices for elementary school-age children. The objective was to investigate the regularity with which SLPs use different types of assessments (described across data types, task types, environmental contexts, and dynamic features). This study also investigated factors that influence assessment practice, the main sources from which SLPs obtain information on language assessment and the main challenges reported by SLPs in relation to language assessment.Method: A web-based survey was used to collect information from 407 Australian SLPs regarding the types of assessments they use. Factors that influenced the regularity with which different types of assessments were used were investigated using regression analysis.Results: Most SLPs regularly used assessments that are norm-referenced, decontextualized, and conducted in a clinical context and less regularly used other types of assessments. Service agency, Australian state, and SLPs' years of experience were found to influence the regularity with which some types of assessments were used. Informal discussions with colleagues were the most frequently identified source of information on assessment practice. Main challenges related to limited time, lack of assessment materials, and lack of confidence in assessing children from culturally and linguistically diverse backgrounds.Conclusions: SLPs could improve current language assessment practice for elementary school-age children through more regular use of some types of assessments. Actions to facilitate evidence-based assessment practice should consider the contextual differences that exist between service agencies and states and address challenges that SLPs experience in relation to language assessment.Otorhinolaryngolog

Individual chitin synthase enzymes synthesize microfibrils of differing structure at specific locations in the Candida albicans cell wall

The shape and integrity of fungal cells is dependent on the skeletal polysaccharides in their cell walls of which β(1,3)-glucan and chitin are of principle importance. The human pathogenic fungus Candida albicans has four genes, CHS1, CHS2, CHS3 and CHS8, which encode chitin synthase isoenzymes with different biochemical properties and physiological functions. Analysis of the morphology of chitin in cell wall ghosts revealed two distinct forms of chitin microfibrils: short microcrystalline rodlets that comprised the bulk of the cell wall; and a network of longer interlaced microfibrils in the bud scars and primary septa. Analysis of chitin ghosts of chs mutant strains by shadow-cast transmission electron microscopy showed that the long-chitin microfibrils were absent in chs8 mutants and the short-chitin rodlets were absent in chs3 mutants. The inferred site of chitin microfibril synthesis of these Chs enzymes was corroborated by their localization determined in Chsp–YFP-expressing strains. These results suggest that Chs8p synthesizes the long-chitin microfibrils, and Chs3p synthesizes the short-chitin rodlets at the same cellular location. Therefore the architecture of the chitin skeleton of C. albicans is shaped by the action of more than one chitin synthase at the site of cell wall synthesis

Energy landscape of relaxed amorphous silicon

We analyze the structure of the energy landscape of a well-relaxed 1000-atom model of amorphous silicon using the activation-relaxation technique (ART nouveau). Generating more than 40,000 events starting from a single minimum, we find that activated mechanisms are local in nature, that they are distributed uniformly throughout the model and that the activation energy is limited by the cost of breaking one bond, independently of the complexity of the mechanism. The overall shape of the activation-energy-barrier distribution is also insensitive to the exact details of the configuration, indicating that well-relaxed configurations see essentially the same environment. These results underscore the localized nature of relaxation in this material.Comment: 8 pages, 12 figure

Inferring superposition and entanglement from measurements in a single basis

We discuss what can be inferred from measurements on one- and two-qubit systems using a single measurement basis at various times. We show that, given reasonable physical assumptions, carrying out such measurements at quarter-period intervals is enough to demonstrate coherent oscillations of one or two qubits between the relevant measurement basis states. One can thus infer from such measurements alone that an approximately equal superposition of two measurement basis states has been created in a coherent oscillation experiment. Similarly, one can infer that a near maximally entangled state of two qubits has been created in an experiment involving a putative SWAP gate. These results apply even if the relevant quantum systems are only approximate qubits. We discuss applications to fundamental quantum physics experiments and quantum information processing investigations.Comment: Final published versio

Rapid Rule-out of Acute Myocardial Infarction With a Single High-Sensitivity Cardiac Troponin T Measurement Below the Limit of Detection: A Collaborative Meta-analysis.

Background: High-sensitivity assays for cardiac troponin T (hs-cTnT) are sometimes used to rapidly rule out acute myocardial infarction (AMI). Purpose: To estimate the ability of a single hs-cTnT concentration below the limit of detection (<0.005 µg/L) and a nonischemic electrocardiogram (ECG) to rule out AMI in adults presenting to the emergency department (ED) with chest pain. Data Sources: EMBASE and MEDLINE without language restrictions (1 January 2008 to 14 December 2016). Study Selection: Cohort studies involving adults presenting to the ED with possible acute coronary syndrome in whom an ECG and hs-cTnT measurements were obtained and AMI outcomes adjudicated during initial hospitalization. Data Extraction: Investigators of studies provided data on the number of low-risk patients (no new ischemia on ECG and hs-cTnT measurements <0.005 µg/L) and the number who had AMI during hospitalization (primary outcome) or a major adverse cardiac event (MACE) or death within 30 days (secondary outcomes), by risk classification (low or not low risk). Two independent epidemiologists rated risk of bias of studies. Data Synthesis: Of 9241 patients in 11 cohort studies, 2825 (30.6%) were classified as low risk. Fourteen (0.5%) low-risk patients had AMI. Sensitivity of the risk classification for AMI ranged from 87.5% to 100% in individual studies. Pooled estimated sensitivity was 98.7% (95% CI, 96.6% to 99.5%). Sensitivity for 30-day MACEs ranged from 87.9% to 100%; pooled sensitivity was 98.0% (CI, 94.7% to 99.3%). No low-risk patients died. Limitation: Few studies, variation in timing and methods of reference standard troponin tests, and heterogeneity of risk and prevalence of AMI across studies. Conclusion: A single hs-cTnT concentration below the limit of detection in combination with a nonischemic ECG may successfully rule out AMI in patients presenting to EDs with possible emergency acute coronary syndrome. Primary Funding Source: Emergency Care Foundation

Epizootic Landscapes: Sheep Scab and Regional Environment in England in 1279–1280

This essay looks at late-medieval rural landscapes of animal disease through the prism of sheep epizootics in England, caused by sheep scab, a highly acute and transmissive disease, whose first wave broke out in 1279–1280. The essay focuses on three regions in England: East Anglia, the Wiltshire-Hampshire Chalklands and Kent, each possessing distinct topographic and environmental features and exhibiting different rates of mortality. The study sets a theoretical model, based on the concept of ‘complexity theory’ and consisting of ten different principles, determining regional variances in dissemination of scab and in mortality patterns. A close analysis of the available statistical sources suggests that there was no ‘universal’ explanatory factor accounting for the correlation between regional geography and mortality rates, and that the situation varied not only from region to region, but from farm to farm, depending on a combination of several possible factors. It is only through a meticulous analysis of local, rather than regional, conditions that the complexity of the situation can begin to be appreciate