Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. Interleukin-6 (IL-6) is involved in the final differentiation of B cells into antibody-producing cells. Interleukin-8 (IL-8) is a neutrophil chemotactic factor that plays an important role in the recruitment of neutrophil to inflammatory loci. Inflammatory mediators by cells in the gingiva have been implicated in the initiation and progression of periodontitis and oral infection. The purpose of this study was conducted to investigate the effect of lipopolysaccharide (LPS), staphylococcus enterotoxin B (SEB) on production of IL-6 and IL-8 by human gingival and facial dermal fibroblasts. Primary cultured human gingival and facial dermal fibroblasts were incubated with LPS (0.01, 0.1, $1.0{\mu}g/ml$), SEB (0.01, 0.1, $1.0{\mu}g/ml$) or LPS $(0.1{\mu}g/ml)$ plus SEB $(0.1{\mu}g/ml)$. Culture supernatants were collected at 24, 48, and 72 hrs and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. IL-6 production in gingival fibroblasts stimulated with LPS was higher than that with SEB. IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-6 production in facial dermal fibroblasts was increased only by stimulation with a high concentration of LPS $(1.0{\mu}g/ml)$. Its production in facial dermal fibroblasts by exposure with SEB was decreased in comparison with control, nontreated cells. Therefore, gingival fibroblasts showed higher sensitivity than facial dermal fibroblasts in response to low concentration of LPS. Also, IL-6 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in gingival fibroblasts was enhanced greatly only by stimulation of high concentration of LPS $(1.0{\mu}g/ml)$. That by exposure with SEB was increased only in 24 hrs cultivation. IL-8 production by double exposure with LPS plus SEB was amplified in comparison with single exposure of LPS or SEB. IL-8 production in facial dermal fibroblasts was decreased by LPS and increased only in 48 hrs cultivation by SEB. IL-8 production by double exposure with LPS plus SEB was enhanced only in 48 hrs cultivation in comparison with single exposure of LPS or SEB. therefore, IL-6 and IL-8 production were released at various quantities according to bacterial toxin applied and site of fibroblast harvested. These results suggest that gingival fibroblasts may be concerned with IL-6 and IL-8 related inflammatory response more than facial dermal fibroblasts.
Proceedings of the Plant Resources Society of Korea Conference
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2019.10a
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pp.8-8
/
2019
The stringent response (SR) is characterized as a bacterial defense mechanism in response to various growth-inhibiting stresses. It is activated by accumulation of a small nucleotide regulator, (p)ppGpp, and induces global changes in bacterial transcription and translation. Recent work from our group has shown that (p)ppGpp plays a critical role in virulence and survival in Vibrio cholerae. The genes, relA and relV, are involved in the production of (p)ppGpp, while the spoT gene encodes an enzyme that hydrolyzes it in V. cholerae. A mutant strain defective in (p)ppGpp production (i.e. ${\Delta}relA{\Delta}relV{\Delta}spoT$ mutant) lost the ability to produce cholera toxin (CT) and lost their viability due to uncontrolled production of organic acids, when grown with extra glucose. In contrast, the ${\Delta}relA{\Delta}spoT$ mutant, a (p)ppGpp overproducer strain, produced enhanced level of CT and exhibited better growth in glucose supplemented media via glucose metabolic switch from organic fermentation to acetoin, a neutral fermentation end product, fermentation. These findings indicates that (p)ppGpp, in addition to its well-known role as a SR mediator, positively regulates CT production and maintenance of growth fitness in V. cholerae. This implicates SR as a promising drug target, inhibition of which may possibly downregulate V. cholerae virulence and survival fitness. Therefore, we screened a chemical library and identified a compound that induces medium acidification (termed iMAC) and thereby loss of wild type V. cholerae viability under glucose-rich conditions. Further, we present a potential mechanism by which the compound inhibits (p)ppGpp accumulation. Together, these results indicate that iMAC treatment causes V. cholerae cells to produce significantly less (p)ppGpp, an important regulator of the bacterial virulence and survival response, and further suggesting that it has a therapeutic potential to be developed as a novel antibacterial agent against cholera.
Seul Gi, Baek;Jin Ju, Park;Sosoo, Kim;Mi-Jeong, Lee;Ji-Seon, Paek;Jangnam, Choi;Ja Yeong, Jang;Jeomsoon, Kim;Theresa, Lee
The Plant Pathology Journal
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v.38
no.6
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pp.637-645
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2022
Fusarium head blight (FHB) is one of the most serious diseases in barley and wheat, as it is usually accompanied by the production of harmful mycotoxins in the grains. To identify FHB-resistant breeding resources, we evaluated 60 elite germplasm accessions of barley (24) and wheat (36) for FHB and mycotoxin accumulation. Assessments were performed in a greenhouse and five heads per accession were inoculated with both Fusarium asiaticum (Fa73, nivalenol producer) and F. graminearum (Fg39, deoxynivalenol producer) strains. While the accessions varied in disease severity and mycotoxin production, four wheat and one barley showed <20% FHB severity repeatedly by both strains. Mycotoxin levels in these accessions ranged up to 3.9 mg/kg. FHB severity was generally higher in barley than in wheat, and Fa73 was more aggressive in both crops than Fg39. Fg39 itself, however, was more aggressive toward wheat and produced more mycotoxin in wheat than in barley. FHB severity by Fa73 and Fg39 were moderately correlated in both crops (r = 0.57/0.60 in barley and 0.42/0.58 in wheat). FHB severity and toxin production were also correlated in both crops, with a stronger correlation for Fa73 (r = 0.42/0.82 in barley, 0.70 in wheat) than for Fg39.
The effects of ginseng saponin and its related materials on aflatoxin production by Aspergillus parasiticus NRRL2999 in yeast extract sucrose (YES) medium were studied. Maximal production of aflatoxins by the mold in the medium occurred after 9 days at $28^{\circ}C$. When various concentrations of ginseng saponin were added to the medium aflatoxin productions were significantly reduced (p<0.05) compared to the control after 9 days at $28^{\circ}C$. 0.05% of saponin in the medium greatly decreased aflatoxin synthesis, and no aflatoxins were synthesized by the mold in the medium contained 5.0% of saponin. When various concentrations of saponin diol and triol were added to the medium both ingibitory and sitimulatory effects on alfatoxin production were resulted. Saponin fraction numbers of 1, 2, 4, 5 and 6 decreased aflatoxin production, however the numbers of 3 and 7 stimulated the toxin production. 0.05% of adenosine, guanosine, caffeine and xanthosine in the media inhibited aflatoxin production (p<0.05), but adenine and cytosine increased the production. When 5.0% of saponin was added to the medium aflatoxins were not synthesized at all, but total lipid synthesis and mold growth were highly stimulated. Both the synthesis of total lipid and mold growth were reduced in case of aflatoxin synthesis stimulated.
Kang Seong Yeob;Kim Sung Hoon;Kim Su Myung;Namgoong Uk;Kim Dong Hee
Journal of Physiology & Pathology in Korean Medicine
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v.18
no.6
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pp.1832-1842
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2004
The present study was performed to examine the possible anti-inflammatory effects of a herbal drug ASCH in RAW264.7 cell line. Inflammation was induced by LPS toxin treatment to RAW264.7 macrophage cell line. Increases in cytokine production such as IL-1β, IL-6. and IL-18, COX-2, NOS-Ⅱ (iNOS), and TNF-alpha were observed at mRNA level in the LPS-treated RAW264.7 cells. Measurement of IL-6, nitric oxide and the reactive oxygen species (ROS) showed increased production of these inflammation mediators. Treatment of ASCH effectively decreased IL-1β protein in a dose-dependent manner, and IL-6 and IL-18 were reduced at 100㎍/㎖ of ASCH concentration. NO production was also decreased by ASCH treatment. A slight inhibition for TNF-alpha in terms of protein, but not mRNA level was obtained by 100㎍/㎖ of ASCH treatment. ASCH treatment to normal RAW264.7 cells did not produce any cytotoxicity, indicting that the action of ASCH was selective to inflammatory cells. Thus, the present data suggest that ASCH may act as an important regulator to alleviate the inflammatory symptoms.
The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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v.24
no.3
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pp.1-16
/
2011
Objective : In the theory of Korean medicine, Scrophulariae Radix (SR) can clear away heat and cool the blood, nourish yin and promote the production of the body fluids, relieve toxin and benefit the throat. The present study was carried out to investigate effects of SR on allergic contact dermatitis (ACD) induced by 2,4-dinitrochlorobenzene (DNCB) in mice. Methods : In this experiment, effects of SR on clinical aspects on the skin, histopathological changes such as spongiosis, mast cell distribution, immune cell infiltration in tissue, spleen / body ratio and production levels of serum cytokines were investigated in vivo. In addition, effects on cell viability and release of b-hexosaminidase and histamine were also investigated in vitro. Results : SR treatment diminished erythema, desquamation and keratosis which were induced by repeated painting of DNCB. Spongiosis and edema were diminished by painting of SR in histopathological observation, infiltrations of mast cell and monocytes were also decreased in SR group. In addition, spleen / body ratio was lowered compared to ADC control group. Production level of IFN-${\gamma}$ in serum was decreased, but level of IL-4 did not affected by SR. Finally, more than 400 ${\mu}g/ml$ of SR treatment groups showed decreased cell viabilities in RBL-2H3 cells. Treatment with over 200 ${\mu}g/ml$ of SR decreased b-hexosaminidase release, and treatment with over 400 ${\mu}g/ml$ decreased histamine release in vitro. Conclusion : these data suggest that SR can decrease symptoms of ACD, then SR is useful to treat patient with ACD.
Lee, Theresa;Lee, Seung-Ho;Shin, Jean Young;Kim, Hee-Kyoung;Yun, Sung-Hwan;Kim, Hwang-Yong;Lee, Soohyung;Ryu, Jae-Gee
The Plant Pathology Journal
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v.30
no.1
/
pp.33-42
/
2014
Nivalenol (NIV) and deoxynivalenol (DON) are predominant Fusarium-producing mycotoxins found in grains, which are mainly produced by Fusarium asiaticum and F. graminearum. NIV is found in most of cereals grown in Korea, but the genetic basis for NIV production by F. asiaticum has not been extensively explored. In this study, 12 genes belonging to the trichothecene biosynthetic gene cluster were compared at the transcriptional level between two NIV-producing F. asiaticum and four DON-producing F. graminearum strains. Chemical analysis revealed that time-course toxin production patterns over 14 days did not differ between NIV and DON strains, excluding F. asiaticum R308, which was a low NIV producer. Both quantitative real-time polymerase chain reaction and Northern analysis revealed that the majority of TRI gene transcripts peaked at day 2 in both NIV and DON producers, which is 2 days earlier than trichothecene accumulation in liquid medium. Comparison of the gene expression profiles identified an NIV-specific pattern in two transcription factor-encoding TRI genes (TRI6 and TRI10) and TRI101, which showed two gene expression peaks during both the early and late incubation periods. In addition, the amount of trichothecenes produced by both DON and NIV producers were correlated with the expression levels of TRI genes, regardless of the trichothecene chemotypes. Therefore, the reduced production of NIV by R308 compared to NIV or DON by the other strains may be attributable to the significantly lower expression levels of the TRI genes, which showed early expression patterns.
Kim, Kyoung-Mi;Kim, Hye-Ju;Ryu, Jae-Ha;Sohn, Dong-Hwan
Journal of Ginseng Research
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v.24
no.2
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pp.79-82
/
2000
Saponins, the major component of ginseng root, mediate the pharmacological action of the ginseng. It has been reported that ginseng roots have protective effect against various toxins. In this study, the effects of ginseng total saponin (GTS) on tumor necrosis factor-alpha (TNF-$\alpha$) production induced by bacterial toxin was investigated. TNF-$\alpha$ level in lipopolysaccharides (LPS)-activated serum was remarkably reduced by intraperitoneal administration (50 mg/kg)of ginseng total saponin (GTS) into mice. The inhibitory effect against TNF-$\alpha$ production was not significant when GTS was given after the LPS injection, and by oral administration. These results suggested that ginseng root may have protective activity against liver damage accompanying the overproduction of TNF-$\alpha$ and GTS is the active component of ginseng.
Codium fragile (CF) is an edible green alga consumed as a traditional food source in Korea. In this study, the ethanol extract of CF was evaluated to determine if it has anti-inflammatory activity. Lipopolysaccharide (LPS), a toxin from bacteria, is a potent inducer of inflammatory cytokines, such as tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Therefore, we studied whether CF extracts have an anti-inflammatory effect in LPS-induced murine macrophage cell lines (RAW 264.7). In the present study, IL-6 production was measured using an enzyme-linked immunosorbent assay (ELISA), prostaglandin $E_2$($PGE_2$) production was measured using the EIA kit, and cyclooxygenase (COX)-2 and mitogen-activated protein kinase (MAPK) activation were determined by Western blot analysis. IL-6 mRNA, COX-2 mRNA and iNOS mRNA expression were measured using reverse transcription-polymerase chain reaction (RT-PCR). The results indicated that CF extracts inhibit LPS-induced IL-6, NO and PGE2 production in a dose-dependent manner, as well as expression of iNOS and COX-2. CF extracts significantly inhibited LPS-induced c-Jun N-terminal kinase (JNK) 1/2 phosphorylation. Taken together, these findings may help elucidate the mechanism by which CF modulates RAW 264.7 cell activation under inflammatory conditions.
The structural components of microorganism are quite related to the toxin and environmental conditions. The vegetative and dormant cells are quite affected by the physical and chemical environments to survive and they will be dormant when they are in the extreme environment. The mechanism to activate the microorganisms however, is not well defined yet in the area of activation state and sporulation state through the analysis of EPS. Other than that even the main mechanism of prior to acquisition of analysis values is not well understood. Therefore, what kind of specific environment to affect the activation and sporulation will be discussed through the analysis of the extracellular polymeric substances(EPS). EPS are a high molecular weight mixture of polymers presenting both outside of cells and interior of microbial aggregates. They are a major complex materials in microbial aggregation for sustaining them together in a three dimensional matrix. Three commonly used extraction methods were applied to this study their effectiveness and quantification in extracting EPS from several Bacillus microorganisms and activated sludge. Three extraction methods used for this study are regular centrifugation with formaldehyde (RCF), Steaming, and EDTA extraction. The results of EPS contents such as the quantitative proteins, carbohydrates and the ratio of protein versus carbohydrate from the several species with the several specific methods showed in this research. This study aims to get comparable results of the quantitative production of EPS and the effectiveness of sedimentation for Bacillus microorganisms and activated sludge from several wastewater treatment plans. The results revealed that the protein amount extracted was the highest by the Steaming method of three extraction methods before sporulation and the carbohydrate amount extracted was the highest by the RCA method of three extraction methods after sporulation. The higher amount of protein compared with carbohydrate from Bacillus microorganisms affected higher sedimentation efficiency, however it could not be found the relation between the EPS production and sedimentation efficiency for the activated sludge.
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