Control of Competence in Vibrio fischeri.

Vibrio species, including the squid symbiont Vibrio fischeri, become competent to take up DNA under specific conditions. For example, V. fischeri becomes competent when grown in the presence of chitin oligosaccharides or upon overproduction of the competence regulatory factor TfoX. While little is known about the regulatory pathway(s) that controls V. fischeri competence, this microbe encodes homologs of factors that control competence in the well-studied V. cholerae To further develop V. fischeri as a genetically tractable organism, we evaluated the roles of some of these competence homologs. Using TfoX-overproducing cells, we found that competence depends upon LitR, the homolog of V. cholerae master quorum-sensing and competence regulator HapR, and upon homologs of putative pilus genes that in V. cholerae facilitate DNA uptake. Disruption of genes for negative regulators upstream of LitR, namely, the LuxO protein and the small RNA (sRNA) Qrr1, resulted in increased transformation frequencies. Unlike LitR-controlled light production, however, competence did not vary with cell density under tfoX overexpression conditions. Analogous to the case with V. cholerae, the requirement for LitR could be suppressed by loss of the Dns nuclease. We also found a role for the putative competence regulator CytR. Finally, we determined that transformation frequencies varied depending on the TfoX-encoding plasmid, and we developed a new dual tfoX and litR overexpression construct that substantially increased the transformation frequency of a less genetically tractable strain. By advancing the ease of genetic manipulation of V. fischeri, these findings will facilitate the rapid discovery of genes involved in physiologically relevant processes, such as biofilm formation and host colonization.IMPORTANCE The ability of bacteria to take up DNA (competence) and incorporate foreign DNA into their genomes (transformation) permits them to rapidly evolve and gain new traits and/or acquire antibiotic resistances. It also facilitates laboratory-based investigations into mechanisms of specific phenotypes, such as those involved in host colonization. Vibrio fischeri has long been a model for symbiotic bacterium-host interactions as well as for other aspects of its physiology, such as bioluminescence and biofilm formation. Competence of V. fischeri can be readily induced upon overexpression of the competence factor TfoX. Relatively little is known about the V. fischeri competence pathway, although homologs of factors known to be important in V. cholerae competence exist. By probing the importance of putative competence factors that control transformation of V. fischeri, this work deepens our understanding of the competence process and advances our ability to genetically manipulate this important model organism.

Development of a Novel Emergency Department Mapping Tool.

OBJECTIVES: Develop a built environment mapping workflow. Implement the workflow in the emergency department (ED). Demonstrate the actionable representations of the data that can be collected using this workflow. BACKGROUND: The design of the healthcare built environment impacts the delivery of patient care and operational efficiency. Studying this environment presents a series of challenges due to the limitations associated with existing technology such as radio-frequency identification. The authors designed a customized mapping workflow to collect high-resolution spatial, temporal, and activity data to improve healthcare environments, with emphasis on patient safety and operational efficiency. METHOD: A large, urban, academic medical center ED collaborated with an architecture firm to create a data collection, and mapping workflow using ArcGIS tools and data collectors. The authors developed tools to collect data on the entire ED, as well as individual patients, physicians, and nurses. Advanced visual representations were created from the master data set. RESULTS: In 48 consecutive hourly snapshots, 5,113 data points were collected on patients, physicians, nurses, and other staff reflecting the operations of the ED. Separately, 84 patients, 10 attending physicians, 10 resident physicians, and 17 nurses were tracked. CONCLUSIONS: The data obtained from this pilot study were used to create advanced visual representations of the ED environment. This cost-effective ED mapping workflow may be applied to other healthcare settings. Further investigation to evaluate the benefits of this high-resolution data is required.

The potential impact of a prophylactic vaccine for Ebola in Sierra Leone.

The 2014 outbreak of Ebola virus disease (EVD) in West Africa was multinational and of an unprecedented scale primarily affecting the countries of Guinea, Liberia, and Sierra Leone. One of the qualities that makes EVD of high public concern is its potential for extremely high mortality rates (up to 90%). A prophylactic vaccine for ebolavirus (rVSV-ZEBOV) has been developed, and clinical trials show near-perfect efficacy. We have developed an ordinary differential equations model that simulates an EVD epidemic and takes into account (1) transmission through contact with infectious EVD individuals and deceased EVD bodies, (2) the heterogeneity of the risk of becoming infected with EVD, and (3) the increased survival rate of infected EVD patients due to greater access to trained healthcare providers. Using fitted parameter values that closely simulate the dynamics of the 2014 outbreak in Sierra Leone, we utilize our model to predict the potential impact of a prophylactic vaccine for the ebolavirus using various vaccination strategies including ring vaccination. Our results show that an rVSV-ZEBOV vaccination coverage as low as 40% in the general population and 95% in healthcare workers will prevent another catastrophic outbreak like the 2014 outbreak from occurring.