SignificanceIn plants, DNA cytosine methylation plays a central role in diverse cellular functions, from transcriptional regulation to maintenance of genome integrity. Vast numbers of whole-genome bisulphite sequencing (WGBS) datasets have been generated to profile DNA methylation at single-nucleotide resolution, yet computational analyses vary widely among research groups, making it difficult to cross-compare findings. Here we reprocessed hundreds of publicly available Arabidopsis WGBS libraries using a uniform pipeline. We identified high-confidence differentially methylated regions and compared libraries using a hierarchical framework, allowing us to identify relationships between methylation pathways. Furthermore, by using a large number of independent wild-type controls, we effectively filtered out spontaneous methylation changes from those that are biologically meaningful.

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Controlling the motion of liquid drops on the solid surface has broad technological implications. In this study, the many-body dissipative particle dynamics (MDPD) was employed to study the drop behaviors on chemical chessboard-patterned surfaces formed by square or triangular tiles. The scaling relationship of the model was established based on the surface tension, viscosity, and density of a real fluid, and an improved contact angle measurement technique was introduced to the MDPD system. For drops on a horizontal plane with different tile sizes, the equilibrium morphology was examined. The critical Bond number, that is, the critical dimensionless force which is required to unpin the drop, was found strongly affected by the size and the shape of the tiles. Once the droplet begins to move, the tile pattern and the size strongly affect the velocity fluctuation while weakly affect the average velocity. Interestingly, besides the common straight forward path, two more route patterns (zigzag and oblique) were observed by only tuning the tile angle, indicating that the advancing routes of the drop may vary according to the tile angle. To the author's knowledge, this phenomenon has not been reported in the literature. This study provides a valuable tool to explore the possibility of passive control of the drop's motion by energy-free chemical heterogeneous surfaces and thus is helpful for engineers to design a surface that could manipulate the drop motion without external energy.

Porcine beta-defensin 2 (PBD-2) which is a member of the family of antimicrobial peptides, is widely expressed in pig organs with a broad spectrum of bactericidal activities confirmed in vitro . We previously demonstrated that transgenic (TG) pigs overexpressing PBD-2 could resist the infection by the porcine pathogen Actinobacillus pleuropneumoniae . In this study, the roles of PBD-2 in protecting against bacterial infection were further investigated. The biochemical indexes of the blood sample, body weights, histological morphologies, and weights of the organs of TG mice expressing PBD-2 were measured. Results confirmed that these mice showed normal physiological features. An assay of Salmonella Typhimurium infection was conducted on wild-type (WT) and TG mice. The TG mice possessed higher survival rate, less body weight loss, and pathological changes and smaller recovery rates of bacteria after infection with S . Typhimurium. The in vitro synthetic PBD-2 and the serum and tissue homogenates from the TG mice displayed a direct bactericidal activity. Moreover, PBD-2 could inhibit the release of the proinflammatory cytokines, including IL-6, TNF-α, IL-1β, and IL-12, in the TG mice infected with S . Typhimurium or treated with lipopolysaccharide (LPS). The WT mice treated with PBD-2 and S . Typhimurium or LPS showed reduced levels of proinflammatory cytokines. The mouse macrophage cell line RAW 264.7 which expressed PBD-2 was constructed to detect the signal pathways affected by PBD-2. The suppressing effect of PBD-2 on the release of the proinflammatory cytokines was confirmed using RAW 264.7 either expressing PBD-2 or supplemented with PBD-2. The promoter activity and mRNA level of NF-κB were detected, and PBD-2 was shown to significantly inhibit the activation of the NF-κB pathway induced by LPS. The direct interaction of PBD-2 with TLR4 was revealed by isothermal titration calorimetry and far-Western blot in vitro and the coimmunoprecipitation of PBD-2 with TLR4 on RAW 264.7 cells. This interaction indicates one reason for the interference of NF-κB activation. Overall, this study showed that PBD-2 protected against bacterial infection through a direct bactericidal activity and alleviated inflammation by interfering with the TLR4/NF-κB pathway.