Nurse mentored, student research in undergraduate nursing education to support evidence-based practice: A pilot study.

OBJECTIVE: The aim was to investigate if an extracurricular research skills development program builds the knowledge, attitudes, and skills (KAS) to support evidence-based practice (EBP). METHODS: Twenty nursing students and six mentors in four teams completed small, student-led research projects over 1 year. Using a mixed-methods design, the knowledge, attitudes, and practice (KAP) survey was administered at three-time points, followed by qualitative interviews. A linear mixed-effects regression model was used to analyze survey data and thematic analysis for qualitative data. RESULTS: The change from the KAP survey from the first to the third time point showed a statistically significant difference following engagement in the program. Qualitative data indicated benefits and challenges to participation for both students and mentors. Mentorship provided students with improved relationships, collaboration, and leadership skills. Students believed the program enhanced their understanding of research and reported increased confidence in using EBP. CONCLUSION: Offering students innovative first-hand experiences with research develops research KAS to support EBP.

Binding of the RNA Chaperone Hfq on Target mRNAs Promotes the Small RNA RyhB-Induced Degradation in Escherichia coli.

Many RNA-RNA interactions depend on molecular chaperones to form and remain stable in living cells. A prime example is the RNA chaperone Hfq, which is a critical effector involved in regulatory interactions between small RNAs (sRNAs) and cognate target mRNAs in Enterobacteriaceae. While there is a great deal of in vitro biochemical evidence supporting the model that Hfq enhances rates or affinities of sRNA:mRNA interactions, there is little corroborating in vivo evidence. Here we used in vivo tools including reporter genes, co-purification assays, and super-resolution microscopy to analyze the role of Hfq in RyhB-mediated regulation, and we found that Hfq is often unnecessary for efficient RyhB:mRNA complex formation in vivo. Remarkably, our data suggest that a primary function of Hfq is to promote RyhB-induced cleavage of mRNA targets by RNase E. Moreover, our work indicates that Hfq plays a more limited role in dictating regulatory outcomes following sRNAs RybB and DsrA complex formation with specific target mRNAs. Our investigation helps evaluate the roles played by Hfq in some RNA-mediated regulation.

Identification of bacterial sRNA regulatory targets using ribosome profiling.

Bacteria express large numbers of non-coding, regulatory RNAs known as 'small RNAs' (sRNAs). sRNAs typically regulate expression of multiple target messenger RNAs (mRNAs) through base-pairing interactions. sRNA:mRNA base-pairing often results in altered mRNA stability and/or altered translation initiation. Computational identification of sRNA targets is challenging due to the requirement for only short regions of base-pairing that can accommodate mismatches. Experimental approaches have been applied to identify sRNA targets on a genomic scale, but these focus only on those targets regulated at the level of mRNA stability. Here, we utilize ribosome profiling (Ribo-seq) to experimentally identify regulatory targets of the Escherichia coli sRNA RyhB. We not only validate a majority of known RyhB targets using the Ribo-seq approach, but also discover many novel ones. We further confirm regulation of a selection of known and novel targets using targeted reporter assays. By mutating nucleotides in the mRNA of a newly discovered target, we demonstrate direct regulation of this target by RyhB. Moreover, we show that Ribo-seq distinguishes between mRNAs regulated at the level of RNA stability and those regulated at the level of translation. Thus, Ribo-seq represents a powerful approach for genome-scale identification of sRNA targets.

Antagonistic functions between the RNA chaperone Hfq and an sRNA regulate sensitivity to the antibiotic colicin.

The RNA chaperone Hfq is a key regulator of the function of small RNAs (sRNAs). Hfq has been shown to facilitate sRNAs binding to target mRNAs and to directly regulate translation through the action of sRNAs. Here, we present evidence that Hfq acts as the repressor of cirA mRNA translation in the absence of sRNA. Hfq binding to cirA prevents translation initiation, which correlates with cirA mRNA instability. In contrast, RyhB pairing to cirA mRNA promotes changes in RNA structure that displace Hfq, thereby allowing efficient translation as well as mRNA stabilization. Because CirA is a receptor for the antibiotic colicin Ia, in addition to acting as an Fur (Ferric Uptake Regulator)-regulated siderophore transporter, translational activation of cirA mRNA by RyhB promotes colicin sensitivity under conditions of iron starvation. Altogether, these results indicate that Fur and RyhB modulate an unexpected feed-forward loop mechanism related to iron physiology and colicin sensitivity.

Dual-acting riboswitch control of translation initiation and mRNA decay.

Riboswitches are mRNA regulatory elements that control gene expression by altering their structure in response to specific metabolite binding. In bacteria, riboswitches consist of an aptamer that performs ligand recognition and an expression platform that regulates either transcription termination or translation initiation. Here, we describe a dual-acting riboswitch from Escherichia coli that, in addition to modulating translation initiation, also is directly involved in the control of initial mRNA decay. Upon lysine binding, the lysC riboswitch adopts a conformation that not only inhibits translation initiation but also exposes RNase E cleavage sites located in the riboswitch expression platform. However, in the absence of lysine, the riboswitch folds into an alternative conformation that simultaneously allows translation initiation and sequesters RNase E cleavage sites. Both regulatory activities can be individually inhibited, indicating that translation initiation and mRNA decay can be modulated independently using the same conformational switch. Because RNase E cleavage sites are located in the riboswitch sequence, this riboswitch provides a unique means for the riboswitch to modulate RNase E cleavage activity directly as a function of lysine. This dual inhibition is in contrast to other riboswitches, such as the thiamin pyrophosphate-sensing thiM riboswitch, which triggers mRNA decay only as a consequence of translation inhibition. The riboswitch control of RNase E cleavage activity is an example of a mechanism by which metabolite sensing is used to regulate gene expression of single genes or even large polycistronic mRNAs as a function of environmental changes.

Small RNA-mediated regulation at the level of transcript stability.

In recent years, bacterial small regulatory RNAs (sRNAs) have been demonstrated to be powerful modulators of gene expression. Whether it is by modulating mRNA functions or protein activities, sRNAs often employ unexpected and extremely diverse mechanisms to modify the genetic output. Although the first sRNAs were characterized as molecules blocking translation of specific target mRNAs, this review will focus on an emerging subset of sRNAs that promote the decay of their target mRNAs. While the outcome resembles the RNAi silencing described in eukaryotes, the mechanism of bacterial sRNAs differs fundamentally. These sRNAs are the subject of intensive studies, which makes them the best characterized sRNAs to date.

Strategies to increase familiarization and acceptance of electronic health records among health professionals and consumers.

There are many reasons an organization may choose to implement electronic health records. The challenge is to acknowledge the benefits and deficiencies of the electronic health record, to understand the driving forces for implementation and the barriers to it, and to effect change in the workplace and consumer behaviour. Indeed, one challenge is the determination of the organizational stance with regard to these factors. If all of these factors are seriously considered and the risks to implementation measured, a successful implementation should ensue.