Effect of LAB on TGF-β induced pro-fibrogenic markers expression in LX-2 cells.

Confluence LX-2 cells were treated with LAB for 48 h, then stimulated with TGF-β for 24 h. The mRNA levels of α-SMA, Col1A1, MMP-2, ICAM-1, TIMP-1, and TIMP2 were determined by RT-PCR. LX-2 cells, either with medium or TGF-β alone, were used as a negative and positive control. Two independent experiments (n = 3) were performed, and the average values (mean + S.D) are shown. *p p p <0.001.</p

LAB strains modulate SMAD-independent TGF-β signaling in LX-2 cells.

Confluence LX-2 cells were treated with LABs for 48 h, and stimulated with TGF-β for 120 minutes. Western blot was performed to determine the phosphorylation of ERK, p38, and IκBα at the indicated time point. The Image J software was used to determine the intensities of protein bands. The bar graphs are the average of two different experiments (mean + S.D). *p p p <0.001.</p

LAB strains modulate the apoptosis-associated markers in LX-2 in response to TGF-β.

Confluence LX-2 cells were treated with LABs for 48 h, then stimulated with TGF-β for 120 minutes. Western blot was performed to determine the level of Bcl-2, Bax, and Caspase 3 at the indicated time point. The Image J software was used to determine the intensities of protein bands. The bar graphs are the average of two different experiments (mean + S.D). *p p p <0.001.</p

LAB reduces the accumulation of collagen in LX-2 cells in response to TGF-β.

(A) The amount of collagen deposition in LX-2 cells was determined after 48 h incubation with LAB strains and with TGF-β for 12 h and 24 h. Two independent experiments (n = 3) were performed, and the average values (mean + S.D) are shown. *p p p L. plantarum+TGF-β are showed.</p

Presentation_1_Immunobiotic Strains Modulate Toll-Like Receptor 3 Agonist Induced Innate Antiviral Immune Response in Human Intestinal Epithelial Cells by Modulating IFN Regulatory Factor 3 and NF-κB Signaling.pptx

Many studies have demonstrated that immunobiotics with immunoregulatory functions improve the outcomes of several bacterial and viral infections by modulating the mucosal immune system. However, the precise mechanisms underlying the immunoregulatory and antiviral activities of immunobiotics have not yet been elucidated in detail. The present study was conducted to determine whether selected lactic acid bacteria (LAB) modulate toll-like receptor 3 (TLR3) agonist polyinosinic:polycytidylic acid (PolyI:C) induced viral response in human intestinal epithelial cells (IECs). PolyI:C increased the expression of interferon-β (IFN-β), interleukin-6 (IL-6), interleukin-8 (IL-8), monocyte chemoattractant protein (MCP-1), and interleukin-1β (IL-1β) in HCT116 cells, and these up-regulations were significantly altered when cells were pre-stimulated with LAB isolated from Korean fermented foods. LAB strains were capable to up-regulate IFN-β but down-regulated IL-6, IL-8, MCP-1, and IL-1β mRNA levels as compared with PolyI: C. HCT-116 cell treatment with LABs beneficially modulated the mRNA levels of C-X-C motif chemokine 10 (CXCL-10), 2-5A oligoadenylate synthetase 1 (OSA1), myxovirus resistance protein (MxA), TLR3, and retinoic acid inducible gene-I (RIG-I), and TLR negative regulators. In addition, LABs increased IFN-β, IFN-α, and interleukin-10 (IL-10) and decreased tumor necrosis factor-α (TNF-α) and IL-1β protein/mRNA levels in THP-1 cells. LABs also protected the cells by maintaining tight-junction proteins (zonula occludens-1 and occludin). The beneficial effects of these LABs were mediated via modulation of the interferon regulatory factor 3 (IRF3) and nuclear factor-kappa B (NF-κB) pathways. Overall, the results of this study indicate that immunobiotics have potent antiviral and anti-inflammatory activities that may use as antiviral substitutes for human and animal applications.</p

Cuboplexes: Topologically Active siRNA Delivery

RNAi technology is currently experiencing a revival due to remarkable improvements in efficacy and viability through oligonucleotide chemical manipulations and/or <i>via</i> their packaging into nanoscale carriers. At present, there is no FDA-approved system for siRNA technology in humans. The design of the next generation of siRNA carriers requires a deep understanding of how a nanoparticle’s physicochemical properties truly impart biological stability and efficiency. For example, we now know that nanoparticles need to be sterically stabilized in order to meet adequate biodistribution profiles. At present, targeting, uptake, and, in particular, endosomal escape are among the most critical challenges impairing RNAi technologies. The disruption of endosomes encompasses membrane transformations (for example, pore formation) that cost significant elastic energy. Nanoparticle size and shape have been identified as relevant parameters impacting tissue accumulation and cellular uptake. In this paper, we demonstrate that the internal structure of lipid-based particles offers a different handle to promote endosomal membrane topological disruptions that enhance siRNA delivery. Specifically, we designed sterically stabilized lipid-based particles that differ from traditional liposomal systems by displaying highly ordered bicontinuous cubic internal structures that can be loaded with large amounts of siRNA. This system differs from traditional siRNA-containing liposomes (lipoplexes) as the particle–endosomal membrane interactions are controlled by elasticity energetics and not by electrostatics. The resulting “PEGylated cuboplex” has the ability to deliver siRNA and specifically knockdown genes with efficiencies that surpass those achieved by traditional lipoplex systems

<i>Lactobacillus</i> strains modulate TNF-α induced β-defensin-2 expression in IECs.

Caco2 cells were pre-stimulated separately with Lactobacillus alone (A) and post-stimulation with TNF-α 12 h (B). The expression of β-defensin-2 expression was analyzed by RT-PCR. Caco2 cells with or without presence of TNF-α were used as controls. The results represented here is an average values (mean ± S.D) of three independent values. Significant differences between the groups, *p<0.05.</p

Effect of LAB on TGF-β induced proinflammatory cytokine/chemokine expression in LX-2 cells.

Confluence LX-2 cells were treated with LAB for 48 h, and stimulated with TGF-β for 24 h. The IL-6, CXCL-8, CCL2, IL-1β, TNF-α, and TRAF6 expression levels were determined by RT-PCR. The LX-2 cells, either with medium or TGF-β alone, were used as the negative and positive control. Two independent experiments (n = 3) were performed, and the average values (mean + S.D) are shown. *p p p <0.001.</p

LAB strains modulate the autophagy-associated markers in LX-2, in response to TGF-β.

Confluence LX-2 cells were treated with LABs for 48 h, and stimulated with TGF-β for 120 minutes. Western blot was performed to determine the level of ATG5, p62, LC3I/II, and p-AKT and p-mTOR at the indicated time point. Image J software was used to determine the intensities of the protein bands. The bar graphs are the average of two different experiments (mean +_S.D). *p p p <0.001.</p