Docosahexaenoic acid differentially modulates plasma membrane targeting and subcellular localization of lipidated proteins in colonocytes

Correct localization of lipidated cytosolic proteins to the plasma membrane (PM) is mediated by interactions between lipid anchors of proteins and cell membranes. Previously, dietary fish oil and its major n-3 polyunsaturated fatty acid (PUFA), docosahexaenoic acid (DHA), have been shown to decrease Ras membrane association, concomitantly reducing rat colon tumor incidence and Ras signaling, compared with corn oil and linoleic acid (LA), a highly prevalent vegetable fat and dietary PUFA in the U.S. diet. In order to explore the potential regulatory role of the cellular lipid environment in PM targeting of lipidated proteins, young adult mouse colon (YAMC) cells were treated with 50 µM DHA, LA, or oleic acid (OA) 24 h prior to and 36-48 h after transfection with green fluorescent protein (GFP) fusion constructs of various lipidated cytosolic proteins. Relative expression of each GFP fusion protein at the PM and the Golgi in living cells was quantified using z-serial confocal microscopy and digital image processing. DHA differentially altered the subcellular localization of Ras isoforms and Src-related tyrosine kinases in a reversible manner. DHA significantly decreased the PM localization and increased the endomembrane association of H-Ras, N-Ras, and Lck, which are targeted to the PM via the exocytic pathway, regardless of their functional state. In contrast, the subcellular distribution of K-Ras and Fyn, of which transport is independent of the vesicular transport pathway, was unaffected by DHA. Moreover, DHA selectively inhibited lipidated cytosolic protein targeting since the PM delivery of transmembrane protein cargo was unaffected, indicating that DHA does not alter the bulk flow of secretory vesicular traffic. Overall, the present study presents compelling evidence that select dietary constituents with membrane lipid-modifying properties can differentially modulate subcellular localization of important lipidated signaling proteins depending on their intracellular trafficking route to the PM

BallGAN: 3D-aware Image Synthesis with a Spherical Background

3D-aware GANs aim to synthesize realistic 3D scenes such that they can be rendered in arbitrary perspectives to produce images. Although previous methods produce realistic images, they suffer from unstable training or degenerate solutions where the 3D geometry is unnatural. We hypothesize that the 3D geometry is underdetermined due to the insufficient constraint, i.e., being classified as real image to the discriminator is not enough. To solve this problem, we propose to approximate the background as a spherical surface and represent a scene as a union of the foreground placed in the sphere and the thin spherical background. It reduces the degree of freedom in the background field. Accordingly, we modify the volume rendering equation and incorporate dedicated constraints to design a novel 3D-aware GAN framework named BallGAN. BallGAN has multiple advantages as follows. 1) It produces more reasonable 3D geometry; the images of a scene across different viewpoints have better photometric consistency and fidelity than the state-of-the-art methods. 2) The training becomes much more stable. 3) The foreground can be separately rendered on top of different arbitrary backgrounds.Comment: Project Page: https://minjung-s.github.io/ballga

Exopolysaccharide of Enterococcus faecium L15 promotes the osteogenic differentiation of human dental pulp stem cells via p38 MAPK pathway

Abstract Background Bone has important functions in the body. Several researchers have reported that the polysaccharides and lipopolysaccharide derived from microbes can promote osteogenic differentiation of stem cells. Enterococcus faecium, a lactic acid bacterium (LAB), produces several bioactive metabolites and has been widely applied in the food and nutraceutical industries. The exopolysaccharide (EPS) from LAB has also been extensively examined for its postbiotic effects and for its in vivo and in vitro functionalities. However, studies on promoting bone differentiation using polysaccharides from LAB are lacking. Therefore, the purpose of this study was to investigate the effect of E. faecium L15 extract and EPS on osteogenic differentiation of human dental pulp stem cells (hDPSCs) and to identify the underlying mechanisms. Methods hDPSCs were obtained from dental pulp tissue, and L15 extract and EPS were isolated from L15. Gene and protein expression of the osteogenic differentiation markers were analyzed with qPCR and western blotting and the possible signaling pathways were also investigated using western blotting. Osteogenic differentiation potential was examined by alkaline phosphatase (ALP) staining and alizarin red s (ARS) staining. In addition, osteogenic differentiation potential of L15 EPS was explored in ex vivo culture of neonate murine calvaria. Results The calcium deposition and ALP activity were enhanced by addition of L15 extract or EPS. The expression levels of RUNX2, ALP, and COL1A1 mRNA and the protein expression levels of RUNX2, ALP, and BMP4 were increased in hDPSCs treated with the L15 extract or EPS. The L15 EPS treatment enhanced phosphorylation of the p38 mitogen-activated protein kinase (MAPK). The L15 EPS-induced increases in RUNX2, ALP, and BMP4 expression were suppressed by the p38 MAPK inhibitor SB203580. The promoting effect of L15 EPS on osteogenic differentiation was not only seen in hDPSCs, but also in osteoblast precursors. ALP activity and the expression of RUNX2, ALP, and COL1A1 increased in the L15 EPS-treated osteoblast precursors. In addition, L15 EPS increased bone thickness of neonate murine calvaria in ex vivo culture. Conclusions The stimulatory effect of L15 extract and EPS on osteogenic differentiation occurred through the p38 MAPK pathway, and L15 EPS enhanced new bone formation in neonate murine calvaria. These data suggest that L15 EPS has therapeutic potential applicable to bone regeneration

The impact of the COVID-19 pandemic and the expansion of free vaccination policy on influenza vaccination coverage: An analysis of vaccination behavior in South Korea.

BackgroundAnnual vaccination for influenza is globally recommended for some prioritized groups due to its high morbidity and mortality. Until 2019, South Korea has provided free influenza vaccination to children aged ≤12, adults aged ≥65, and pregnant women to enhance vaccination coverage. In 2020, with the COVID-19 pandemic, free flu vaccination was temporarily broadened to adults aged 62-64 and children aged 13-18. We analyzed the trends in influenza vaccination coverages in South Korea and evaluated the impact of the COVID-19 pandemic and the expansion of the free vaccination policy on influenza vaccination coverage.MethodsWe conducted a cross-sectional study with nationwide survey data from Korea National Health and Nutrition Examination Survey (KNHANES). We evaluated the trends in influenza vaccination coverages of target populations from 2010 to 2020. Influenza vaccination coverages of children aged 13-18, adults aged 62-64, and adults aged ≥65 were compared between 2019 and 2020.ResultsIn total, 72,443 individuals were analyzed. From 2019 to 2020, with the expansion of free influenza vaccination and the COVID-19 pandemic, the vaccination coverage of children aged 13-18 increased from 27.8% to 43.5% (PConclusionIn 2020, along with COVID-19 outbreaks, a decline of influenza vaccination coverage in older adults was observed regardless of free immunizations. It is likely due to behavioral changes to reduce the risk of COVID-19 transmission. This is supported by a greater reduction of influenza vaccination coverage in regions with higher COVID-19 outbreaks, as well as by South Korea's high medical accessibility and highly congested medical facilities. To sustain a high level of vaccination coverage of high-risk population during epidemics, additional efforts beyond free vaccination policies should be implemented

Cellular internalization effect of Ara27 in various cell lines

Protein and peptide candidates are screened to apply therapeutic application as a drug. Ensuring that these candidates are delivered and maximized effectiveness is still challenging and a variety of studies are ongoing. As drug delivery system vehicles, cell-penetrating peptide (CPP) can deliver various kinds of cargo into the cell cytosol. In a previous study, we developed Ara27 CPP, which are a zinc knuckle family protein of Arabidopsis, and confirmed internalization in human dermal fibroblasts and human dental pulp stem cells at low concentration with short time treatment condition without any toxicity. Ara27, an amphipathic CPP, could be modified and utilized in the biomedical field excluding the risk of toxicity. Therefore, we would like to confirm the non-toxic induced penetrating ability of Ara27 in various cell lines. The purpose of this study was to screen the cell internalization ability of Ara27 in various cell lines and to confirm Ara27 as a promising core CPP structure. First, Ara27 was screened to confirm non-toxicity concentration. Then, fluorescence-labeled Ara27 was treated on human normal cell lines, cancer cell lines and animal cell lines to identify the cellular internalization of Ara27. Ara27 was well intracellular localized in all cell lines and the intensity of fluorescence was remarkably increased in time pass manner. These results indicate that Ara27 has the potential as a core structure for applications in various drug delivery systems

Experimental and Simulation Studies on Adsorption and Diffusion Characteristics of Coalbed Methane

Coalbed methane (CBM) content is generally estimated using the isotherm theory between pressure and adsorbed amounts of methane. It usually determines the maximum content of adsorbed methane or storage capacity. However, CBM content obtained via laboratory experiment is not consistent with that in the in-situ state because samples are usually ground, which changes the specific surface area. In this study, the effect of the specific surface area relative to CBM content was investigated, and diffusion coefficients were estimated using equilibrium time analysis. The differences in adsorbed content with sample particle size allowed the determination of a specific surface area where gases can adsorb. Also, there was an equilibrium time difference between fine and lump coal, because more time is needed for the gas to diffuse through the coal matrix and adsorb onto the surface in lump coal. Based on this, we constructed a laboratory-scale simulation model, which matched with experimental results. Consequently, the diffusion coefficient, which is usually calculated through canister testing, can be easily obtained. These results stress that lump coal experiments and associated simulations are necessary for more reliable CBM production analysis

Comparison of Cell-Free Extracts from Three Newly Identified Lactobacillus plantarum Strains on the Inhibitory Effect of Adipogenic Differentiation and Insulin Resistance in 3T3-L1 Adipocytes

Obesity and associated metabolic disorders, including cardiovascular disease and diabetes, are rapidly becoming serious global health problems. It has been reported that Lactobacillus plantarum (L. plantarum) extracts have the beneficial activities of antiobesity and antidiabetes, although few studies have compared the beneficial effects among various L. plantarum extracts. In this study, three new L. plantarum (named LP, LS, and L14) strains were identified, and the antiobesogenic and diabetic effects of their extracts were investigated and compared using 3T3-L1 cells in vitro. Lipid accumulation in maturing 3T3-L1 cells was significantly decreased by the addition of LS and L14 extracts. The mRNA expression levels of Pparγ, C/ebpα, Fabp4, Fas, and Dgat1 were significantly decreased by the addition of LP, LS, and L14 extracts. Interestingly, the protein expression levels of PPARγ, C/EBPα, FABP4, and FAS were downregulated in mature 3T3-L1 cells with the addition of the L14 extract. Moreover, the LS and L14 extract treatments stimulated glucose uptake in maturing adipocytes. The L14 extract treatments exhibited a significant reduction in TNF-α protein expression, which is a key factor of insulin resistance in adipocytes. Of the three extracts, L14 extract markedly reduced adipogenic differentiation and insulin resistance in vitro, suggesting that the L14 extract may be used as a therapeutic agent for obesity-associated metabolic disorders

Single-Temporal Sentinel-2 for Analyzing Burned Area Detection Methods: A Study of 14 Cases in Republic of Korea Considering Land Cover

Forest fires are caused by various climatic and anthropogenic factors. In Republic of Korea, forest fires occur frequently during spring when the humidity is low. During the past decade, the number of forest fire incidents and the extent of the damaged area have increased. Satellite imagery can be applied to assess damage from these unpredictable forest fires. Despite the increasing threat, there is a lack of comprehensive analysis and effective strategies for addressing these forest fires, particularly considering the diverse topography of Republic of Korea. Herein, we present an approach for the automated detection of forest fire damage using Sentinel-2 images of 14 areas affected by forest fires in Republic of Korea during 2019–2023. The detection performance of deep learning (DL), machine learning, and spectral index methods was analyzed, and the optimal model for detecting forest fire damage was derived. To evaluate the independent performance of the models, two different burned areas exhibiting distinct characteristics were selected as test subjects. To increase the classification accuracy, tests were conducted on various combinations of input channels in DL. The combination of false-color RNG (B4, B8, and B3) images was optimal for detecting forest fire damage. Consequently, among the DL models, the HRNet model achieved excellent results for both test regions with intersection over union scores of 89.40 and 82.49, confirming that the proposed method is applicable for detecting forest fires in diverse Korean landscapes. Thus, suitable mitigation measures can be promptly designed based on the rapid analysis of damaged areas