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

Contribution of actin filaments and microtubules to cell elongation and alignment depends on the grating depth of microgratings

Additional file 1: Figure S1. (A) A phase contrast image of TCPS surface. Bar, 100 μm. (B) An imageshowing FN-lines (1 μm line and spacing) obtained by Atomic Force Microscopy (AFM) (Dimension 3100with a Nanoscope III controller, Digital Instruments) using silicon cantilevers (spring constant; 50 Nm-1)(RTESP, Veeco Probes) in contact mode. (C-E) SEM (Scanning electron microscopy) (6010 LV, JEOL)images showing the cross section of three different microgratings; 1 μm gratings with 0.35 um depth (C) and1 μm depth (D) and 2 μm gratings with 2 μm depth (E). Figure S2. (A) Fluorescence image of a RPE-1 cell stably expressing GFP/centrin cell on 1 μm gratings (1 μm deep). Bar, 30 μm. A yellow arrow indicates the direction of cell elongation. (B) Average cell aspect ratio (R) of cells on 1 μm gratings (0.35 or 1 μm deep) and 2 μm gratings with/without CD treatment. n: number of cells. ***P < 0.001. Data were analyzed using one-way ANOVA and a Bonferroni post hoc test. Error bar denotes the standard deviation of the mean. Figure S3. Alignment of actin and vinculin to the different substrates (Flat TCPS surface, FN-lines, and 1 μm gratings (0.35 or 1μm deep)). The alignment angle was measured as an angle difference of actin or vinculin orientation to the long axis of a cell on flat PDMS surface or the long axis of the FN-line or each micrograting. #: the number of cells. Error bar denotes the standard deviation of the mean. Figure S4. Merged image of MTs (Green fluorescence) and pattern (phase contrast) of cells on 1 μm grating (1 μm deep) in the presenceof CD at 1 μM

Cellular plasticity and immune microenvironment of malignant pleural effusion are associated with EGFR-TKI resistance in non-small-cell lung carcinoma

Malignant pleural effusion (MPE) is a complication of lung cancer that can be used as an alternative method for tissue sampling because it is generally simple and minimally invasive. Our study evaluated the diagnostic potential of non-small-cell lung carcinoma (NSCLC)-associated MPE in terms of understanding tumor heterogeneity and identifying response factors for EGFR tyrosine kinase inhibitor (TKI) therapy. We performed a single-cell RNA sequencing analysis of 31,743 cells isolated from the MPEs of 9 patients with NSCLC (5 resistant and 4 sensitive to EGFR TKI) with EGFR mutations. Interestingly, lung epithelial precursor-like cells with upregulated GNB2L1 and CAV1 expression were enriched in the EGFR TKI-resistant group. Moreover, GZMK upregulated transitional effector T cells, and plasmacytoid dendritic cells were significantly enriched in the EGFR TKI-resistant patients. Our results suggest that cellular plasticity and immunosuppressive microenvironment in MPEs are potentially associated with the TKI response of patients with EGFR-mutated NSCLC

Performance Estimation Method of Li-Ion Starting-Lighting-Ignition Batteries of Electric Vehicle through Lab-Scale Pouch Cell Experiment

CCA (cold-cranking amps) is used as one of the standards to define the power of a SLI (starting-lighting-ignition) battery of electric vehicles (EVs) by applying a large discharge current for a short time. It is easier to start an engine at a high temperature than a low temperature, so tests are usually conducted at a low temperature to meet the demands of users. However, in a large cell, the internal temperature of the cell rises due to the heat of the cell when a large current is applied. This phenomenon can increase the overall cell power performance because it has the effect of reducing ohmic overvoltage and improving electrode kinetics. Therefore, it is difficult to distinguish the effect of the temperature rise and applied current on the voltage curve during the consecutive pulse discharge. In this study, three times of 10 C discharge current pulse (~ 7 mA/cm2 in this study) is applied on a cell for 30 seconds, with rest periods of 600s, 30s included between each pulse. Evaluations were conducted from -18 ??? to 50 ???, with a LiFePO4/graphite lab-scale pouch cell rather than an actual large cell. Therefore, the temperature of the cell remained constant under the evaluation conditions in which heat generation is almost zero, so the voltage curves can be interpreted by excluding the effect of the temperature rise of a cell. Based on these experimental results, voltage curves were simulated depending on each internal resistance that related to Joule heating of the actual large cell. It is possible to predict the performance and degree of temperature rise of large cells. Also, this study proposes an effective and economical method to distinguish the effect of the temperature rise of the large cell and the applied current by using a lab-scale pouch cell with controlled temperature

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

Oral intake of Lactobacillus plantarum

Objectives Whether periodic oral intake of postbiotics positively affects weight regulation and prevents obesity-associated diseases in vivo is unclear. This study evaluated the action mechanism of Lactobacillus plantarum L-14 (KTCT13497BP) extract and the effects of its periodic oral intake in a high-fat-diet (HFD) mouse model. Materials and methods Mouse pre-adipocyte 3T3-L1 cells and human bone marrow mesenchymal stem cells (hBM-MSC) were treated with L-14 extract every 2 days during adipogenic differentiation, and the mechanism underlying anti-adipogenic effects was analysed at cellular and molecular levels. L-14 extract was orally administrated to HFD-feeding C57BL/6J mice every 2 days for 7 weeks. White adipose tissue was collected and weighed, and liver and blood serum were analysed. The anti-adipogenic mechanism of exopolysaccharide (EPS) isolated from L-14 extract was also analysed using Toll-like receptor 2 (TLR2) inhibitor C29. Results L-14 extract inhibited 3T3-L1 and hBM-MSC differentiation into mature adipocytes by upregulating AMPK signalling pathway in the early stage of adipogenic differentiation. The weight of the HFD + L-14 group (31.51 +/- 1.96 g) was significantly different from that of the HFD group (35.14 +/- 3.18 g). L-14 extract also significantly decreased the serum triacylglycerol/high-density lipoprotein cholesterol ratio (an insulin resistance marker) and steatohepatitis. In addition, EPS activated the AMPK signalling pathway by interacting with TLR2, consequently inhibiting adipogenesis. Conclusions EPS from L-14 extract inhibits adipogenesis via TLR2 and AMPK signalling pathways, and oral intake of L-14 extract improves obesity and obesity-associated diseases in vivo. Therefore, EPS can be used to prevent and treat obesity and metabolic disorders.Y

Psychological well-being and salivary markers of inflammation: The moderating effect of age

© 2022 International Association of Applied Psychology.Increasing evidence suggests a significant impact of higher psychological well-being (PWB) on health outcomes; however, such associations have been studied exclusively in middle-aged to older adults. This study examined the aging effect on PWB measures as well as the moderating effect of age on the link between PWB and inflammation, using salivary markers by comparing the younger adults (n = 127; Mage = 22.98 years) versus older adults (n = 75; Mage = 75.60 years). Older adults showed significantly lower levels of PWB, particularly regarding purpose in life and personal growth. Moreover, higher purpose in life was associated with lower salivary IL-1ß and IL-6 (b = 0.83, p <.001; b = 0.81, p <.01) only in the older adult group but not in younger adults. These findings highlight the potential buffering effect of the sense of living well on physiological pathways in later life.11Nssciscopu

Seasonal Compositions of Size-Fractionated Surface Phytoplankton Communities in the Yellow Sea

Little information on the phytoplankton community in the Yellow Sea (YS)—especially size-fractionated phytoplankton—is currently available, in comparison to the various physicochemical studies in the literature. Using high-performance liquid chromatography (HPLC), size-fractionated phytoplankton communities were seasonally investigated in the YS in 2019. In the study period, diatoms (55.0 ± 10.2%) and cryptophytes (16.9 ± 9.3%) were the dominant groups. Due to the recent alteration in inorganic nutrient conditions reported in the YS, the contribution of diatoms was lower than in previous studies. The large-sized phytoplankton group (>20 µm) was dominated mostly by diatoms (89.0 ± 10.6%), while the small-sized phytoplankton group (<20 µm) was also dominated by diatoms (41.9 ± 9.1%), followed by cryptophytes (19.2 ± 9.8%). The contributions of small-sized diatoms (<20 µm) have been overlooked in the past, as they are difficult to detect, but this study confirms significant amounts of small-sized diatoms, accounting for 62.3% of the total diatoms in the YS. This study provides an important background for assessing the seasonal variations in different-sized diatom groups in the YS. Further detailed studies on their potential ecological roles should be conducted, in order to better understand marine ecosystems under future warming scenarios