Biochemical characterization of a recombinant Japanese encephalitis virus RNA-dependent RNA polymerase

<p>Abstract</p> <p>Background</p> <p>Japanese encephalitis virus (JEV) NS5 is a viral nonstructural protein that carries both methyltransferase and RNA-dependent RNA polymerase (RdRp) domains. It is a key component of the viral RNA replicase complex that presumably includes other viral nonstructural and cellular proteins. The biochemical properties of JEV NS5 have not been characterized due to the lack of a robust <it>in vitro </it>RdRp assay system, and the molecular mechanisms for the initiation of RNA synthesis by JEV NS5 remain to be elucidated.</p> <p>Results</p> <p>To characterize the biochemical properties of JEV RdRp, we expressed in <it>Escherichia coli </it>and purified an enzymatically active full-length recombinant JEV NS5 protein with a hexahistidine tag at the N-terminus. The purified NS5 protein, but not the mutant NS5 protein with an Ala substitution at the first Asp of the RdRp-conserved GDD motif, exhibited template- and primer-dependent RNA synthesis activity using a poly(A) RNA template. The NS5 protein was able to use both plus- and minus-strand 3'-untranslated regions of the JEV genome as templates in the absence of a primer, with the latter RNA being a better template. Analysis of the RNA synthesis initiation site using the 3'-end 83 nucleotides of the JEV genome as a minimal RNA template revealed that the NS5 protein specifically initiates RNA synthesis from an internal site, U₈₁, at the two nucleotides upstream of the 3'-end of the template.</p> <p>Conclusion</p> <p>As a first step toward the understanding of the molecular mechanisms for JEV RNA replication and ultimately for the <it>in vitro </it>reconstitution of viral RNA replicase complex, we for the first time established an <it>in vitro </it>JEV RdRp assay system with a functional full-length recombinant JEV NS5 protein and characterized the mechanisms of RNA synthesis from nonviral and viral RNA templates. The full-length recombinant JEV NS5 will be useful for the elucidation of the structure-function relationship of this enzyme and for the development of anti-JEV agents.</p

The Impact of Overactive Bladder on Health-Related Quality of Life, Sexual Life and Psychological Health in Korea

Purpose We aimed to estimate the prevalence of overactive bladder (OAB) in Korea, to assess the variation in prevalence by sex and age, and to measure the impact of OAB on quality of life. Methods A population-based, cross-sectional telephone survey was conducted between April and June 2010 with a questionnaire regarding the prevalence of OAB, demographics, and the impact of OAB on quality of life. A geographically stratified random sample of men and women aged ≥30 years was selected. Results The overall prevalence of OAB was 22.9% (male, 19%; female, 26.8%). Of a total of 458 participants with OAB, 37.6% and 19.9% reported moderate or severe impact on their daily life and sexual life (5.6% and 3.5%, respectively, in participants without OAB). Anxiety and depression were reported by 22.7% and 39.3% of participants with OAB, respectively (9.7% and 22.8%, respectively, in participants without OAB). Only 19.7% of participants with OAB had consulted a doctor for their voiding symptoms, but 50.7% of respondents with OAB were willing to visit a hospital for the management of their OAB symptoms. Conclusions This study confirmed that OAB symptoms are highly prevalent in Korea, and many sufferers appear to have actively sought medical help. OAB has severe effects on daily and sexual life as well as psychological health

Selection of Elevation Models for Flood Inundation Map Generation in Small Urban Stream: Case Study of Anyang Stream

To reduce flood damages, the Ministry of Environment in Korea has provided a flood inundation map so that people can expediently identify flood-prone areas. However, the current flood inundation maps have been produced based on the DEM which makes it difficult to represent realistic situations due to the lack of reproduction of land surface conditions. This study aims to provide more accurate and detailed flood inundation maps for flooding events due to river overflow in small urban areas. In this study, flood inundation analysis is performed using the river analysis system, HEC-RAS 2D, with the DSM and the DEM of urban areas in the Anyang Stream Basin, Korea to examine the differences in terms of terrain data and flooded area. Finally, for urban areas with dense buildings and congested road networks, the flood inundation analysis based on DSM can represent a more realistic flood situation and create an appropriate flood inundation map

Hydraulic and ecological changes under drainage gate operations with coupled model SCHISM-CoSiNE in Saemangeum basin, Korea

The drainage gates have been controlled for desalination under normal conditions and flood defense in&nbsp; Saemangeum basin, Korea. Recently, it became an issue that the gates have been opened not to deteriorate water quality in the lake. It is, thus, necessary to precisely estimate the changes of water quality characteristics, especially DO, phosphate and nitrate, in the lake according to various gate operations. In this study, Semi-implicit Cross-scale Hydroscience Integrated System Model and Carbon, Silicate, Nitrogen Ecosystem model (SCHISM-CoSiNE) which is cable to simulate dynamic exchange such as gate operation conditions was utilized to obtain reliable and reasonable results including hydrodyanamic and environmental variables. For the verification, the measured data at 6 locations in Saemangeum basin was used to compare&nbsp; incluidng temperatue and salintiy from 2016 and each relative error became small enough to show high accurary. Also, under various scenarios by changing the designated water surface elevation on flood seasons, this model has been applied to present the best designated water surface elevation in terms of both water quality and water supply in the Saemangeum basin. It becomes possbile to show reliable guidance for dynamic operations and environmental changes with this model as requested in near future

Topological Semantic Graph Memory for Image-Goal Navigation

A novel framework is proposed to incrementally collect landmark-based graph memory and use the collected memory for image goal navigation. Given a target image to search, an embodied robot utilizes semantic memory to find the target in an unknown environment. % The semantic graph memory is collected from a panoramic observation of an RGB-D camera without knowing the robot's pose. In this paper, we present a topological semantic graph memory (TSGM), which consists of (1) a graph builder that takes the observed RGB-D image to construct a topological semantic graph, (2) a cross graph mixer module that takes the collected nodes to get contextual information, and (3) a memory decoder that takes the contextual memory as an input to find an action to the target. On the task of image goal navigation, TSGM significantly outperforms competitive baselines by +5.0-9.0% on the success rate and +7.0-23.5% on SPL, which means that the TSGM finds efficient paths. Additionally, we demonstrate our method on a mobile robot in real-world image goal scenarios

Characterization of Curli A Production on Living Bacterial Surfaces by Scanning Probe Microscopy

AbstractCurli are adhesive surface fibers produced by many Enterobacteriaceae, such as Escherichia coli and Salmonella enterica. They are implicated in bacterial attachment and invasion to epithelial cells. In this study, atomic force microscopy was used to determine the effects of curli on topology and mechanical properties of live E. coli cells. Young's moduli of both curli-deficient and curli-overproducing mutants were significantly lower than that of their wild-type (WT) strain, while decay lengths of the former strains were higher than that of the latter strain. Surprisingly, topological images showed that, unlike the WT and curli-overproducing mutant, the curli-deficient mutant produced a large number of flagella-like fibers, which may explain why the strain had a lower Young's modulus than the WT. These results suggest that the mechanical properties of bacterial surfaces are greatly affected by the presence of filamentous structures such as curli and flagella