lncRNA BC200 is processed into a stable Alu monomer.

The non-coding RNA BC200 is elevated in human cancers and is implicated in translation regulation as well as cell survival and proliferation. Upon BC200 overexpression, we observed correlated expression of a second, smaller RNA species. This RNA is expressed endogenously and exhibits cell-type dependent variability relative to BC200. Aptamer tagged expression constructs confirmed that the RNA is a truncated form of BC200, and sequencing revealed a modal length of 120 nt, thus, we refer to the RNA fragment as BC120. We present methodology for accurate and specific detection of BC120 and establish that BC120 is expressed in several normal human tissues and is also elevated in ovarian cancer. BC120 exhibits remarkable stability relative to BC200 and is resistant to knock-down strategies that target the 3' unique sequence of BC200. Combined knock-down of BC200 and BC120 exhibits greater phenotypic impacts than knock-down of BC200 alone and overexpression of BC120 negatively impacts translation of a GFP reporter providing insight into a potential translational regulatory role for this RNA. The presence of a novel, truncated, and stable form of BC200 adds complexity to the investigation of this non-coding RNA that must be considered in future studies of BC200 and other related Alu RNAs.

Nuclear SRP9/SRP14 heterodimer transcriptionally regulates 7SL and BC200 RNA expression.

The SRP9/SRP14 heterodimer is a central component of signal recognition particle (SRP) RNA (7SL) processing and Alu retrotransposition. In this study, we sought to establish the role of nuclear SRP9/SRP14 in the transcriptional regulation of 7SL and BC200 RNA. 7SL and BC200 RNA steady-state levels, rate of decay, and transcriptional activity were evaluated under SRP9/SRP14 knockdown conditions. Immunofluorescent imaging, and subcellular fractionation of MCF-7 cells, revealed a distinct nuclear localization for SRP9/SRP14. The relationship between this localization and transcriptional activity at 7SL and BC200 genes was also examined. These findings demonstrate a novel nuclear function of SRP9/SRP14 establishing that this heterodimer transcriptionally regulates 7SL and BC200 RNA expression. We describe a model in which SRP9/SRP14 cotranscriptionally regulate 7SL and BC200 RNA expression. Our model is also a plausible pathway for regulating Alu RNA transcription and is consistent with the hypothesized roles of SRP9/SRP14 transporting 7SL RNA into the nucleolus for posttranscriptional processing, and trafficking of Alu RNA for retrotransposition.

Predicting human RNA quadruplex helicases through comparative sequence approaches and helicase mRNA interactome analyses.

RNA quadruplexes are non-canonical nucleic acid structures involved in several human disease states and are regulated by a specific subset of RNA helicases. Given the difficulty in identifying RNA quadruplex helicases due to the multifunctionality of these enzymes, we sought to provide a comprehensive in silico analysis of features found in validated RNA quadruplex helicases to predict novel human RNA quadruplex helicases. Using the 64 human RNA helicases, we correlated their amino acid compositions with subsets of RNA quadruplex helicases categorized by varying levels of evidence of RNA quadruplex interaction. Utilizing phylogenetic and synonymous/non-synonymous substitution analyses, we identified an evolutionarily conserved pattern involving predicted intrinsic disorder and a previously identified motif. We analyzed available next-generation sequencing data to determine which RNA helicases directly interacted with predicted RNA quadruplex regions intracellularly and elucidated the relationship with miRNA binding sites adjacent to RNA quadruplexes. Finally, we performed a phylogenetic analysis of all 64 human RNA helicases to establish how RNA quadruplex detection and unwinding activity may be conserved among helicase subfamilies. This work furthers the understanding of commonalities between RNA quadruplex helicases and provides support for the future validation of several human RNA helicases.

Evaluation of Technical Competency in Healthcare Simulation (E-TeCHS) tool: a modified Delphi study.

Background: Graduates of simulation fellowship programmes are expected to have the ability to perform a variety of simulation specific skills at the time of graduation. Currently, simulation fellowship directors have access to tools to assess the ability of a fellow to debrief learners. However, there is no tool to assess a simulation fellow's competency in technical skills. The purpose of our manuscript was to develop and obtain content validation of a novel instrument designed to assess a simulation fellow's ability to perform the five core simulation technical skills. Methods: The study protocol was based on a methodology for content validation of curriculum consensus guidelines. This approach involves a three-step process, which includes the initial delineation of the curricular content. This was then followed by the validation of the curricular content using survey methodology and lastly obtaining consensus on modifications using Delphi methodology. Results: Two rounds of modified Delphi methodology were performed. Seventy-four respondents provided feedback on the round 1 survey and 45 respondents provided feedback on round 2. The final assessment tool has five elements and 16 subitems with four optional subitems. Conclusion: The Evaluation of Technical Competency in Healthcare Simulation tool provides an instrument developed from a national consensus of content experts. This tool provides simulation fellowship directors a method to evaluate fellows' competency in technical skills.