No abstract

Griffithsin (GRFT) is a broad-spectrum antiviral protein that is effective against several glycosylated viruses. Here, we have evaluated the in vitro and in vivo antiviral activities of GRFT against Japanese encephalitis virus (JEV) infection. In vitro experiments showed that treatment of JEV with GRFT before inoculation of BHK-21 cells inhibited infection in a dose-dependent manner, with 99 % inhibition at 100 μg/ml and a 50 % inhibitory concentration (IC(50)) of 265 ng/ml (20 nM). Binding assays suggested that binding of GRFT to JEV virions inhibited JEV infection. In vivo experiment showed that GRFT (5 mg/kg) administered intraperitoneally before virus infection could completely prevent mortality in mice challenged intraperitoneally with a lethal dose of JEV. Our study also suggested that GRFT prevents JEV infection at the entry phase by targeting the virus. Collectively, our data demonstrate that GRFT is an antiviral agent with potential application in the development of therapeutics against JEV or other flavivirus infections.

Progress in wound healing is primarily quantified by the rate of change of the wound's surface area. The most recent guidelines of the Wound Healing Society suggest that a reduction in wound size of <40% within 4 weeks necessitates a reevaluation of the treatment. However, accurate measurement of wound size is challenging due to the complexity of a chronic wound, the variable lighting conditions of examination rooms, and the time constraints of a busy clinical practice. In this paper, we present our methodology to quantify a wound boundary and measure the enclosed wound area reproducibly. The method derives from a combination of color-based image analysis algorithms, and our results are validated with wounds in animal models and human wounds of diverse patients. Images were taken by an inexpensive digital camera under variable lighting conditions. Approximately 100 patient images and 50 animal images were analyzed and a high overlap was achieved between the manual tracings and the calculated wound area by our method in both groups. The simplicity of our method combined with its robustness suggests that it can be a valuable tool in clinical wound evaluations. The basic challenge of our method is in deep wounds with very small surface areas where color-based detection can lead to erroneous results and which could be overcome by texture-based detection methods. The authors are willing to provide the developed MATLAB code for the work discussed in this paper.

Japanese encephalitis virus is one of the most common causes for epidemic viral encephalitis in humans and animals. Herein we demonstrated that cellular helicase DDX3 is involved in JEV replication. DDX3 knockdown inhibits JEV replication. The helicase activity of DDX3 is crucial for JEV replication. GST-pulldown and co-immunoprecipitation experiments demonstrated that DDX3 could interact with JEV non-structural proteins 3 and 5. Co-immunoprecipitation and confocal microscopy analysis confirmed that DDX3 interacts and colocalizes with these viral proteins and viral RNA during the infection. We determined that DDX3 binds to JEV 5' and 3' un-translated regions. We used a JEV-replicon system to demonstrate that DDX3 positively regulates viral RNA translation, which might affect viral RNA replication at the late stage of virus infection. Collectively, we identified that DDX3 is necessary for JEV infection, suggesting that DDX3 might be a novel target to design new antiviral agents against JEV or other flavivirus infections.

High-temperature Daqu, also called Jiang-flavor Daqu, is the saccharifying and fermenting agent for brewing Jiang-flavor Baijiu. During the spontaneous solid-state fermentation of high-temperature Daqu, variations in temperature and moisture lead to microbial diversity and various metabolites, contributing to the different colors of high-temperature Daqu (customarily referred to as white Daqu, black Daqu, yellow Daqu, and red Daqu in production). We aimed to investigate the differences in microbial communities, physicochemical indices, and potential functions among different high-temperature Daqu with different colors (labeled as QW, QB, QY, and QR) by amplicon sequencing. We found that Kroppenstedtia, Bacillus, and Thermoascus were predominant in all samples; Saccharopolyspora and Thermomyces were predominant in QB and QR; and Unclassfied_O_Eurotiales were predominant in QY. The results on the physicochemical characteristics indicated that compared with other Daqu samples, QW exhibited higher protease activity and lower acidity, whereas QB showed the opposite results. QR had the highest esterification yield, and QY exhibited the highest saccharification but lowest esterification yield. Functional prediction demonstrated that the higher abundances of genes encoding bacterial enzymes of QW and QY were related to the considerably higher abundances of Kroppenstedtia in QW (59%) and QY (87%), respectively. The highest abundance of Thermomyces in QB (80%) contributed to the highest abundance of genes encoding fungal enzymes in QB. This study revealed the microbial and functional dissimilarities of color-based high-temperature starters and helped facilitate the liquor fermentation process.