Selaginella tamariscina ( S. tamariscina ) (Beauv.) Spring (Selaginellaceae) has been used in oriental medicine for the treatment of dysmenorrhea, chronic hepatitis, hyperglycemia, amenorrhea, hematuria, prolapse of the anus and metrorrhagia. This study aimed to investigate the synergistic antibacterial activity with existing antimicrobial agents against oral pathogen. The synergistic effects of 50% ethanol extract of S. tamariscina (STE) were evaluated against oral bacteria, either alone or with antibiotics, via broth microdilution and time-kill method. The minimal inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) values for STE, ampicillin and gentamicin against all the tested bacteria ranged between 6.25-50/50-200 μg/mL, 0.0625-16/0.25-32 μg/mL, and 4-128/16-256 μg/ mL, respectively. STE in combination with ampicillin showed a strong synergistic effect against oral bacteria (fractional inhibitory concentration (FIC) index ≤0.5), whereas on combining with gentamicin, it reduced the on half-eighth times than used alone (FICI ≤ 0.5). Furthermore, a time-kill study showed that the growth of the tested bacteria was completely attenuated after 2-6 h of treatment with 1/2 MIC of STE with 1/2 MIC of antibiotics resulted from an increase of the rate of killing in units of CFU/mL to a greater degree than was observed with alone. The results of this study demonstrate the antimicrobial and synergistic activity of STE and antibiotics against oral pathogens.

The roots of Paeonia lactiflora Pallas are important crude drugs in Korean traditional medicine.A decoction of the dried root has been used in the treatment of rheumatoid arthritis, systemic lupus erythematosus, dysmenorrhea, hepatitis, muscle cramping, inflammation, spasms, and fever and stimulate blood circulation.In this study, the combination effect of the ethanol extract of Paeonia lactiflora (PLE) was evaluated against oral bacteria, either alone or with antibiotics, via broth dilution method and checkerboard and time kill assay.In these results, MIC/MBC values for PLE against all the tested bacteria ranged between 250-2000/250-2000 microg/mL, for ampicillin 0.031-16/0.125-32microg/mL, gentamicin 2-256/4-512 microg/mL, erythromycin 0.008-32/0.016-64microg/mL, and vancomycin 0.5-64/1-128 microg/mL, respectively.Furthermore, the MIC and MBC were reduced to one half-four as a result of the combination of PLE with antibiotics.Six hours of treatment with 1/2 MIC of PLE with 1/2 MIC of antibiotics resulted from an increase of the rate of killing in units of CFU/mL to a greater degree than was observed with alone.These results suggest that the ethanol extract of Paeonia lactiflora (PLE) is important in the antibacterial actions of oral pathogen agents.

Porcine epidemic diarrhea virus (PEDV) is a contagious coronavirus infecting pigs that leads to significant economic losses in the swine industry. Given that PEDV infection occurs in gut epithelial cells mainly via the fecal-oral route, induction of PEDV-specific immune responses in the mucosal compartment is required for protective immunity against viral infection. However, an effective mucosal vaccine against the currently prevalent PEDV strain is not available. In this study, we demonstrated that the N-terminal domain (NTD) of the spike (S) protein of PEDV represents a new vaccine candidate molecule to be applied via the mucosal route. We first established an Escherichia coli expression system producing the partial NTD (NTD231-501) of the PEDV S protein. Orally administered NTD231-501 protein specifically interacted with the apical area of M cells in the follicle-associated epithelium of Peyer's patch. Additionally, the NTD protein induced antigen-specific immune responses in both the systemic and mucosal immune compartments when administered orally. Collectively, we propose the NTD of the PEDV S protein to be a candidate mucosal vaccine molecule.

Drynaria fortunei (D. fortunei), widely used in traditional Korean medicine, is reportedly effective in treating inflammation, hyperlipidemia, bone fractures, oxidative damage, arteriosclerosis, rheumatism, and gynecological diseases. The objective of this study was to evaluate the antibacterial effects of the chloroform fraction of D. fortunei (DFCF) and assess the synergistic effects of DFCF with antibiotics against bacterial pathogens. This was carried out by calculating the minimal inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) and performing checkerboard dilution test and time-kill assays. The MICs/MBCs for DFCF, ampicillin, and gentamicin against all oral strains were >39-2,500/5,000 μg/mL, 0.25-64/0.25-64 µg/mL, and 0.5-256/1-512 µg/mL, respectively. DFCF exhibited the highest activity against the periodontic pathogens Prevotella intermedia and Porphylomonas gingivalis. DFCF in combination with ampicillin showed a strong synergistic effect against oral bacteria (fractional inhibitory concentration (FIC) index ≤0.5), whereas on combining with gentamicin, it reduced the on half-eighth times than used alone (FICI ≤ 0.5). DFCF combined with ampicillin or gentamicin killed 100% of most tested bacteria within 3-4 h. The results of this study demonstrate the antimicrobial and synergistic activity of DFCF and antibiotics against oral pathogens.

The antibacterial activity of an extract and several fractions of Sophora flavescens (S. flavescens) root alone and in combination with antibiotics against oral bacteria was investigated by checkerboard assay and time-kill assay. The minimum inhibitory concentration/minimum bactericidal concentration (MIC/MBC) values for all examined bacteria were 0.313-2.5/0.625-2.5 μg/mL for the n-BuOH fraction, 0.625-5/1.25-10 μg/mL for the EtOAc fraction, 0.25-8/0.25-16 μg/mL for ampicillin, 0.5-256/1-512 μg/mL for gentamicin, 0.008-32/0.016-64 μg/mL for erythromycin, and 0.25-64/0.5-128 μg/mL for vancomycin. The n-butanol (n-BuOH) and ethyl acetate (EtOAc) fractions exhibited stronger antibacterial activity against oral bacteria than other fractions and extracts. The MICs and MBCs were reduced to between one half and one quarter when the n-BuOH and EtOAc fractions were combined with antibiotics. After 24 h of incubation, combination of 1/2 MIC of the n-BuOH fraction with antibiotics increased the degree of bactericidal activity. The present results suggest that n-BuOH and EtOAc extracts of S. flavescens root might be applicable as new natural antimicrobial agents against oral pathogens.

Background: Sophora flavescens AITON (Leguminosae) is a typical traditional Korean medical herb considered to exhibit antibacterial, anti-inflammatory, and antipyretic effects, and is also used for the treatment of skin and mucosal ulcers, sores, diarrhea, gastrointestinal hemorrhage, arrhythmia, and eczema.Objective and design: This study examined the inhibitory effects of sophoraflavanone G (SF) of S. flavescens on the bacterial fibrillar protein, Antigen I/II (AgI/II)-N recombinant protein isolated from Streptococcus mutans(rAg I/II)-induced production of nitric oxide (NO) and prostaglandin E2 (PGE(2)). The investigation was focused on whether SF could inhibit the production of proinflammatory mediators such as nitric oxide (NO) and prostaglandin (PG) E2 as well as the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-a, interleukin (IL)-6, nuclear factor (NF)-kappa B and mitogen-activated protein kinases (MAPKs) in rAgI/II-stimulated RAW 264.7 cells using Griess reagent, Enzyme linked immunosorbent assay (ELISA), and Western blotting analysis.Results: SG significantly inhibited the production of NO and PGE(2) and pro-inflammatory cytokines, such as interleukin (IL)-1 beta, IL-6, and tumor necrosis factor alpha in Ag I/II-N-stimulated RAW264.7 cells, which were mediated by the down-regulation of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. The SF inhibited the phosphorylation of I kappa B-alpha, nuclear translocation of p65, and subsequent activation of NF-kappa B in the rAgI/II-stimulated cells. In addition, the SF suppressed the rAgI/II-stimulated activation of ERK MAPK as well as the MAPK inhibitor significantly reduced the rAgI/II-induced production of NO and PGE(2).Conclusion: Collectively, we suggest that the SF inhibits the expression and production of inflammatory mediators by blocking the ERK MAPK mediated pathway and inhibiting the activation of NF-kappa B. (C) 2016 Elsevier Ltd. All rights reserved.

Studies have been focused on natural products with antibacterial and anti-inflammatory activities, such as fucoidan. Many in vivo studies have evaluated the effect of fucoidan on tumor growth, diabetes, obesity, ischemia reperfusion, and oxidative stress. However, the effects of fucoidan on bacteria-induced gingival inflammation and periodontitis have not been reported. We previously characterized the anti-inflammatory effect of fucoidan in vitro. Here, we confirmed the anti-inflammatory activity of fucoidan in a macrophage cell line in terms of its inhibition of the expression of inflammatory mediators and pro-inflammatory cytokines. Additionally, we confirmed the ability of fucoidan to inhibit gingival inflammation, expression of pro-inflammatory cytokines, and neutrophil recruitment in the gingival tissue of mice injected with LPS prepared from P. gingivalis. Interestingly, however, fucoidan did not inhibit the expression of pro-inflammatory cytokines in a P. gingivalis-infected mouse model of periodontitis. Additionally, fucoidan treatment did not lead to clearance of P. gingivalis or improvement of P. gingivalis infection-mediated bone loss in the periodontitis model. We conclude that fucoidan exerts anti-inflammatory effects in vitro and in vivo, together with a limited antibacterial effect in vivo.

Ethnopharmacological relevance: Water extract of Raphanus sativus L (RSL) seeds was traditionally used to treat digestive inflammatory complaints in Korean culture. RSL seeds exerted antioxidant, anti-inflammatory, and anti-septic functions, suggesting their pharmacological potential for the treatment of inflammatory pathologies associated with oxidative stress such as inflammatory bowel disease.Aim of this study: We evaluated the intestinal anti-inflammatory effects of RSL seed water extract (RWE) in experimental rat models of trinitrobenzenesulphonic acid (TNBS)- or dextran sodium sulfate (DSS)-induced colitis.Materials and methods: RWE was characterized by determining the content of sinapic acid as a reference material and then assayed in the DSS and TNBS models of rat colitis. Male Sprague-Dawley rats were divided into 10 groups (n=7/group): non-colitic control, DSS or TNBS control, DSS colitis groups treated with RWE (100 mg/kg) or mesalazine (25 mg/kg), and TNBS colitis groups treated with various doses (10, 40, 70, and 100 mg/kg) of RWE or mesalazine (25 mg/kg). RWE or mesalazine treatment started the same day of colitis induction and rats were sacrificed 24 h after the last treatment followed by histological and biochemical analyses.Results: Oral administration with RWE suppressed intestinal inflammatory damages in both DSS- and TNBS-induced colitic rats. The treatment with 100 mg/kg RWE recovered intestinal damages caused by TNBS or DSS to levels similar to that of mesalazine, decreasing the activity of myeloperoxidase activity and the secretion of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1 beta. RWE treatment inhibited malondialdehyde production and glutathione reduction in colon of colitis rats. The administration of RWE at dose of 100 mg/kg also suppressed the TNBS- or DSS-stimulated expression of TNF-alpha, IL-1 beta, monocyte chemotactic protein-1, inducible nitric oxide, and intercellular adhesion molecule-1. Furthermore, RWE inhibited p38 kinase and DNA nuclear factor-kappa B binding activities, both of which were stimulated in the colitic rats.Conclusions: The current findings show that RWE ameliorates intestinal oxidative and inflammatory damages in DSS and TNBS models of rat colitis, suggesting its beneficial use for the treatment of intestinal inflammatory disorders. (C) 2015 Elsevier Ireland Ltd. All rights reserved.

본 연구의 ëª©ì ì€ Streptococcus mutans의 Ag I/II와 glucosyltransferase에 대해 개발된 ë‹¨ì¼í´ë¡ í•­ì²´ë¥¼ 사용한 타액 내 Streptococcus mutans 검출의 효용성을 평가하기 위함이다. 세 êµ°(성인, ë¯¸ì„±ë „ìž, ê·¸ë¦¬ê³ êµì •ì¹˜ë£Œì¤‘ì¸ ë¯¸ì„±ë „ìž)을 대상으로 DMFT, Dentocult®-SM strip mutans를 이용한 타액내의 colony density (CFU/mL), polymerase chain reaction을 이용한 Sreptococcus mutans와 Sreptococcus sobrinus의 존재여부, ê·¸ë¦¬ê³ 4가지 ë‹¨ì¼í´ë¡ í•­ì²´ë¥¼ 사용한 타액 내 Streptococcus mutans 검출 실험을 시행하였다. ê·¸ ê²°ê³¼ Streptococcus mutans가 Streptococcus sobrinus보다 치아 우식 형성에 더 주요하게 작용한다는 것을 ìž ì¦í•˜ì˜€ìœ¼ë©°, glucosyltransferases (gtf B, gtf C, gtf D)와 Ag I/II에 대한 ë‹¨ì¼í´ë¡ í•­ì²´ë“¤ì˜ Streptococcus mutans를 검출해 내는 ëŠ¥ë ¥ì´ Dentocult®-SM strip mutans나 polymerase chain reaction을 사용한 검출법보다 뛰어남을 보였다.

The human antimicrobial peptide LL‐37 is known to have chemotactic and modulatory activities on various cells including monocytes, T cells, and epithelial cells. Given that LL‐37 enhances chemotactic attraction and modulates the activity of DCs, it is conceivable that it might play a role as an immune adjuvant by skewing the immune environment toward immunostimulatory conditions. In this study, we characterized the mucosal adjuvant activity of LL‐37 using model and pathogenic Ags. When LL‐37‐conjugated Ag was administered orally to mice, a tolerogenic Peyer's patch environment was altered to cell populations containing IL‐6‐secreting CD11c+, CD11c+CD70+, and Th17 cells capable of evoking a subsequent LL‐37‐conjugated Ag‐specific immune response in both systemic and mucosal immune compartments. In addition, we showed presentation of formyl peptide receptor, an LL‐37 receptor, on M cells, which may aid the initiation of an LL‐37‐mediated enhanced immune response through targeting and transcytosis of the conjugated Ag. Based on our findings, we conclude that LL‐37 has potential as an oral mucosal adjuvant, not only by enhancing the delivery of LL‐37‐conjugated Ag to M cells, but also by triggering T‐cell‐mediated Ag‐specific immune responses through modulation of the mucosal immune environment.

There are controversial findings regarding the roles of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)/heme oxygenase-1 (HO-1) pathway on bone metabolism under oxidative stress. We investigated how Nrf2/HO-1 pathway affects osteoblast differentiation of MC3T3-E1 cells in response to hydrogen peroxide (H2O2), N-acetyl cysteine (NAC), or both. Exposing the cells to H2O2 decreased the alkaline phosphatase activity, calcium accumulation, and expression of osteoblast markers, such as osteocalcin and runt-related transcription factor-2. In contrast, H2O2 treatment increased the expression of Nrf2 and HO-1 in the cells. Treatment with hemin, a chemical HO-1 inducer, mimicked the inhibitory effect of H2O2 on osteoblast differentiation by increasing the HO-1 expression and decreasing the osteogenic marker genes. Pretreatment with NAC restored all changes induced by H2O2 to near normal levels in the cells. Collectively, our findings suggest that H2O2-mediated activation of Nrf2/HO-1 pathway negatively regulates the osteoblast differentiation, which is inhibited by NAC.

It is known that formyl peptide receptor-2 (FPR-2) acts as innate immune sensor through recognition of peptides derived from pathogens such as H. pylori and HIV, mitochondrial peptides, amyloidogenic peptide including serum amyloid A, and inflammatory response-associated peptides such as LL-37. However, the role of FPR-2-expressing cells in mucosal immune inductive area, Peyer’s patch (PP), is still unraveled yet. In this study, we identified the expression of FPR-2 in both M cells located in follicle-associated epithelium of PP and follicular dendritic cells (FDCs) localized in germinal center. In addition, the level of Cxcl13 and TGF-β gene transcripts in FDCs, which are closely associated with recruitment and differentiation of IgA-secreting B cells was substantially increased by treatment of cathelicidin LL-37, one of FPR-2 ligands. Moreover, enhanced immune induction was verified in vivo mouse system through monitoring the antigen-specific mucosal immunity after oral administration of LL-37-conjugated antigen. Treatment of LL-37 has also shown the immune modulatory effects such as recruitment of CD11c+ cells and IL-17-secreting cells together with activation of FPR-2-expressing cells. Collectively, this study suggests that FPR-2-expressing cells are associated with regulation of mucosal immune induction and LL-37 could be used as a modulator for the cells.

Recombinant COMP-Ang1, a chimera of angiopoietin-1 (Ang1) and a short coiled-coil domain of cartilage oligomeric matrix protein (COMP), is under consideration as a therapeutic agent capable of inducing the homing of cells with increased angiogenesis. However, the potentials of COMP-Ang1 to stimulate migration of mesenchymal stem cells (MSCs) and the associated mechanisms are not completely understood. We examined the potential of COMP-Ang1 on bone marrow (BM)-MSCs, human periodontal ligament stem cells (PDLSCs), and calvarial osteoblasts. COMP-Ang1 augmented Tie-2 induction at protein and mRNA levels and increased proliferation and expression of runt-related transcription factor 2 (Runx2), osterix, and CXCR4 in BMMSCs, but not in osteoblasts. The COMP-Ang1-mediated increases were inhibited by Tie-2 knockdown and by treating inhibitors of phosphoinositide 3-kinase (PI3K), LY294002, or p38 mitogen-activated protein kinase (MAPK), SB203580. Phosphorylation of p38 MAPK and Akt was prevented by siRNA-mediated silencing of Tie-2. COMP-Ang1 also induced in vitro migration of BMMSCs and PDLSCs. The induced migration was suppressed by Tie-2 knockdown and by CXCR4-specific peptide antagonist or LY294002, but not by SB203580. Furthermore, COMP-Ang1 stimulated the migration of PDLSCs into calvarial defect site of rats. Collectively, our results demonstrate that COMP-Ang1-stimulated proliferation, differentiation, and migration of progenitor cells may involve the Tie-2-mediated activation of p38 MAPK and PI3K/Akt pathways.

Cryptotanshinone (CT), a major tanshinone of medicinal plant Salvia miltiorrhiza Bunge, demonstrated effective in vitro antibacterial activity against all oral bacteria tested in this experiment. The antibacterial activities of CT against oral bacteria were assessed using the checkerboard and time-kill methods to evaluate the synergistic effects of treatment with ampicillin or gentamicin. The CT was determined against oral pathogenic bacteria with MIC and MBC values ranging from 0.5 to 16 and 1 to 64 μg/mL; for am- picillin from 0.0313 to 16 and 0.125 to 32 μg/mL; for gentamicin from 2 to 256 and 4 to 512 μg/mL respectively. The range of MIC50 and MIC90 were 0.0625 - 8 μg/mL and 1 - 64 μg/mL, respectively. The combination effects of CT with antibiotics were synergistic (FIC index Streptococcus sobrinus, S. criceti, and Actinobacillus actinomycetemcomitans (FIC index . Furthermore, a time-kill study showed that the growth of the tested bacteria was completely attenuated after 3 - 6 h of treatment with the 1/2 MIC of CT, regardless of whether it was administered alone or with ampicillin or gentamicin. The results suggest that CT could be employed as a natural antibacterial agent against cariogenic and periodontopathogenic bac- teria.

Actinobacillus pleuropneumoniae is an infective agent that leads to porcine pleuropneumonia, a disease that causes severe economic losses in the swine industry. Based on the fact that the respiratory tract is the primary site for bacterial infection, it has been suggested that bacterial exclusion in the respiratory tract through mucosal immune induction is the most effective disease prevention strategy. ApxIIA is a vaccine candidate against A. pleuropneumoniae infection, and fragment #5 (aa. 439-801) of ApxIIA contains the major epitopes for effective vaccination. In this study, we used mice to verify the efficacy of intranasal immunization with fragment #5 in the induction of protective immunity against nasal challenge with A. pleuropneumoniae and compared its efficacy with that of subcutaneous immunization. Intranasal immunization of the fragment induced significantly higher systemic and mucosal immune responses measured at the levels of antigen-specific antibodies, cytokine-secreting cells after antigen exposure, and antigen-specific lymphocyte proliferation. Intranasal immunization not only efficiently inhibited the bacterial colonization in respiratory organs, but also prevented alveolar tissue damage in infectious condition similar to that of a contaminated pig. Moreover, intranasal immunization with fragment #5 provided acquired protective immunity against intranasal challenge with A. pleuropneumoniae serotype 2. In addition, it conferred cross-protection against serotype 5, a heterologous pathogen that causes severe disease by ApxI and ApxII secretion. Collectively, intranasal immunization with fragment #5 of ApxIIA can be considered an efficient protective immunization procedure against A. pleuropneumoniae infection.

BACKGROUND:Fucoidan is a sulphated polysaccharide that is primarily extracted from brown seaweeds; it has been broadly studied in recent years due to its numerous biological properties, including anticoagulant, antithrombotic, antitumour and antiviral activities. OBJECTIVE AND DESIGN:In this study, fucoidan was evaluated against oral bacteria, either alone or with antibiotics, via the broth dilution method and chequerboard and time-kill assay. RESULTS:Minimum inhibitory concentration/minimum bactericidal concentration (MIC/MBC) values for the fucoidan against all the tested bacteria ranged between 0.125 and 0.50/0.25 and 1.00mgml(-1), for ampicillin 0.125 and 64/0.5 and 64μgml(-1) and for gentamicin 2 and 256/4 and 512μgml(-1), respectively. Furthermore, the MIC and MBC were reduced to one half-eighth as a result of the combination of the fucoidan with antibiotics. One to 3h of treatment with MIC50 of fucoidan with MIC50 of antibiotics resulted from an increase of the rate of killing in colony forming units (CFUs) ml(-1) to a greater degree than was observed with alone. CONCLUSION:These results suggest that fucoidan is important in the antibacterial actions of the agents.

Abstract The mucosal epithelium is exposed to enormous amount of microorganisms containing commensal and pathogenic bacteria. Although many pathogens initiate their infection in mucosal epithelium and/or M cells of Peyer’s patch, detailed mechanism for the infection is poorly understood. We explored the possible survival mechanism of Yersinia enterocolitica in M cells in relation to crosstalk between C5aR and TLRs based on the observation that outermembrane protein H (OmpH) of Y. enterocolitica is one of the ligands for C5aR. Localization of C5aR and TLRs in human M-like cells and mouse M cells was confirmed during Y. enterocolitica infection. Also, regulation of cAMP- and/or autophagy-associated gene expression by Y. enterocolitica infection was characterized. Moreover, C5aR antagonist treatment reduced the survival of Y. enterocolitica in human M-like cells and the bacteria showed low infection efficiency in C5aR knock-out mouse. It is suggested that the OmpH of Y. enterocolitica could be used as M cell-targeting ligand in oral mucosal vaccination model. Collectively, we assume that the inhibition of cAMP expression by interaction between Y. enterocolitica and both C5aR and TLRs is closely associated with the bacterial survival in M cells and this interaction could be used as targeting of oral mucosal vaccine.