• Title/Summary/Keyword: culture supernatants

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Isolation, Identification and Optimal Cultrul Condition of Antioxidant Producing Bacterium Isolated from the Marine Sources

  • Kim, Man-Chul;Heo, Moon-Soo
    • 한국생물공학회:학술대회논문집
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    • 2005.10a
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    • pp.343-346
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    • 2005
  • The isolated strain, SC2-1 was Gram-positive, catalase positive, facultatively anaerobic, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and sodium chloride required for the bacteria growth. The radical scavenging activity of the culture supernatants was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) method. This bacterium was identified based on cellular fatty acids analysis and 16S rDNA sequencing then named Exiguobacterium sp. SC2-1. The optimum culture conditions for production of antioxidant were $25^{\circ}C,$ pH 7.8 and NaCl concentration were 4%. The modified optimal medium compositions were maltose 2.5% (w/v), yeast extract 1.5% (w/v) and $KH_2PO_4$ 0.05% (w/v). Free radical scavenging activity of under optimal culture conditions were 93%.

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Saxatilin Suppresses Tumor-induced Angiogenesis by Regulating VEGF Expression in NCI-H460 Human Lung Cancer Cells

  • Jang, Yoon-Jung;Kim, Dong-Seok;Jeon, Ok-Hee;Kim, Doo-Sik
    • BMB Reports
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    • v.40 no.3
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    • pp.439-443
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    • 2007
  • Tumor growth and metastasis are dependent on angiogenesis, and endothelial cell invasion and migration are apparent means of regulating tumor progression. We report here that saxatilin, a snake venom-derived disintegrin, suppresses the angiogenesis-inducing properties of NCI-H460 human lung cancer cells. Culture supernatants of NCI-H460 cells are able to induce human umbilical vascular endothelial cell (HUVEC) invasion and tube formation. However, treatment of the cancer cells with saxatilin resulted in reduced angiogenic activity of the culture supernatant. This suppressed angiogenic property was found to be associated with the level of vascular endothelial growth factor (VEGF) in the culture supernatant. Further experimental evidence indicated that saxatilin inhibits VEGF production in NCI-H460 cells by affecting hypoxia induced factor-1$\alpha$ (HIF-1$\alpha$) expression via the Akt pathway.

Antagonism of Bacterial Extracellular Metabolites to Freshwater-Fouling Invertebrate Zebra Mussels, Dreissena polymopha

  • Gu, Ji-Dong;Ralph Mitchell
    • Journal of Microbiology
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    • v.39 no.2
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    • pp.133-138
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    • 2001
  • We investigated the antagonism of indigenous bacteria isolated from stressed mussels and their extracellular metabolites on the adult zebra mussel, Dreissena polymorpha. Selective bacterial isolates including Aeromonas media, A. salmonicida, A. veronii, and Shewanella putrefaciens, showed strong lethality against adult mussels and 100% mortality was observed within 5 days of incubation. Bacterial metabolites, fractionated and concentrated from stationary-phase culture supernatants of these bacterial isolates, displayed varying degrees of antagonistic effects on zebra mussels. Among the three size fractions examined, <5, 5-10, and >10 kDa, the mast lethal fraction seems to be >10 kDa for three of the four isolates tested. Further chemical analyses of these size fractions revealed that the predominant constituents were polysaccharides and proteins. No 2-keto-3-deoxyoctanoic acid (2-KDO), deoxyribonucleic acids (DNA) or uranic acid were detectable. Extraction of supernatants of two antagonistic isolates with polar solvent suggested that polar molecules are present in the active fraction. Our data suggest that extracellular metabolites produced by antagonistic bacteria are also involved in disease development in zebra mussels and elucidation of the mechanisms involved may offer a novel strategy for control of biofouling invertebrates.

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Regulatory Effect of Atopic Allergic Reaction by Chungsangbangpoong-Tang (청상방풍탕(淸上防風湯)의 아토피성 알레르기 반응(反應) 조절(調節) 효과(效果))

  • Lim, Tae-Hyung;Park, Eun-Jung;Lee, Hai-Ja;Han, Kyeung-Hoon;Na, Won-Kyeung;Jin, Kyong-Son
    • The Journal of Pediatrics of Korean Medicine
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    • v.18 no.2
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    • pp.61-76
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    • 2004
  • Objective : To demonstrate of regulatory effect of atopic allergic regulation by Chungsangbangpoong-Tang(CBT), This experiment was studied. Methods : The author investigated a possible effect of CBT on cytokines production using human T cell line (MOLT-4) or human mast cell line (HMC-1). In addition, the author investigated whether CBT has inhitory effects on compound 48/80- induced histamine release from rat peritoneal mast cells (RPMC) and compound 48/80-induced ear swelling in ICR mice. Results : CBT (0.01 mg/ml)-containing medium in stimulated culture supernatants significantly increased IL-2 secretion compared with untreated MOLT-4, whereas CBT (0.01-1.0 mg/ml)-containing medium in stimulated culture supernatants significantly decreased IL-4 secretion compared with untreated MOLT-4. Significant reduced levels of IL -6 and $TNF-{\alpha}$ were observed in the HMC-l with CBT (P<0.05). CBT did not inhibit the histamine release from the RPMC but inhibit ear swelling response. Conclusion : These results suggest that CBT contributes to the treatment of atopic allergic reactions, such as atopic dermatitis and that its action may be due to regulation of cytokine production.

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Effects of Omega-3-Rich Harp Seal Oil on the Production of Pro-Inflammatory Cytokines in Mouse Peritoneal Macrophages

  • Choi, Myungwon;Ju, Jaehyun;Suh, Jae Soo;Park, Kun-Young;Kim, Kwang Hyuk
    • Preventive Nutrition and Food Science
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    • v.20 no.2
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    • pp.83-87
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    • 2015
  • Omega-3, a polyunsaturated fatty acid, is an essential fatty acid necessary for human health, and it protects against cardiovascular disease, inflammation, autoimmune diseases, and cancer. In the present study, we investigated the effects of omega-3-rich harp seal oil (HSO) on the production of nitric oxide (NO) and cytokines, such as tumor necrosis factor (TNF)-${\alpha}$, interleukin-(IL)-$1{\beta}$, IL-6, and IL-12/IL-23 (p40) in peritoneal macrophages of mice. The culture supernatants of murine macrophages exposed to lipopolysaccharide (LPS), HSO, or HSO+LPS were harvested to assay IL-$1{\beta}$, TNF-${\alpha}$, IL-6, and IL-12/IL-23 (p40) cytokines and NO. TNF-${\alpha}$, IL-$1{\beta}$, and IL-12/IL-23 (p40) levels, except IL-6, were lower in the culture supernatants of mouse peritoneal macrophages exposed to LPS plus HSO than those of the groups exposed to LPS alone. These observations demonstrate that omega-3-rich harp seal oil downregulates the production of the pro-inflammatory cytokines such as IL-$1{\beta}$, TNF-${\alpha}$, and IL-12/IL-23 (p40). These results suggest that HSO could be potentially used as a preventive agent or as an adjunct in anti-inflammatory therapy, if more research results were accumulated.

Effects of Eucalyptus and Geranium on Production of IL-2 and IL-4 in Mouse Splenocytes (Eucalyptus와 geranium이 마우스 splenocytes에서 IL-2 및 IL-4 생성에 대한 효과)

  • Cha Bong Kyu;Chang Myung Woong;Jeong Young Kee;Kim Kwang Hyuk
    • Journal of Life Science
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    • v.16 no.1
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    • pp.162-167
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    • 2006
  • Aromatherpy is the controlled use of essential oils to promote health and well-being. In this work we have investigated the effect of eucalyptus and geranium on the production of interleukin-2 (IL-2) and interleukin4 (IL-4). Mouse splenocytes were incubated with essential oils. The culture supernatants of mouse splenocytes exposed with these oils were harvested to assay IL-2 and IL-4 production. The quantitative changes of IL-2 in splenocytes culture supernatants after exposure with these oils were decreased at high doses, but increased at low doses. But its of IL-4 were increased generally at high doses of eucalyptus. In case of the exposure of geranium, its of IL-4 were dose-dependently increased. These kinds of essential oils showed the probability to improve IL-2- and IL-4-related immune responses at the optimum exposure.

Bifidobacterium bifidum DS0908 and Bifidobacterium longum DS0950 Culture-Supernatants Ameliorate Obesity-Related Characteristics in Mice with High-Fat Diet-Induced Obesity

  • M. Shamim Rahman;Youri Lee;Doo-Sang Park;Yong-Sik Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.1
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    • pp.96-105
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    • 2023
  • Probiotic supplements have promising therapeutic effects on chronic diseases. In this study, we demonstrated the anti-obesity effects of two potential probiotics, Bifidobacterium bifidum DS0908 (DS0908) and Bifidobacterium longum DS0950 (DS0950). Treatment with DS0908 and DS0950 postbiotics significantly induced the expression of the brown adipocyte-specific markers UCP1, PPARγ, PGC1α, PRDM16 and beige adipocyte-specific markers CD137, FGF21, P2RX5, and COX2 in C3H10T1/2 mesenchymal stem cells (MSCs). In mice with high-fat diet (HFD)-induced obesity, both potential probiotics and postbiotics noticeably reduced body weight and epididymal fat accumulation without affecting food intake. DS0908 and DS0950 also improved insulin sensitivity and glucose use in mice with HFD-induced obesity. In addition, DS0908 and DS0950 improved the plasma lipid profile, proved by reduced triglyceride, low-density lipoprotein, and cholesterol levels. Furthermore, DS0908 and DS0950 improved mitochondrial respiratory function, confirmed by the high expression of oxidative phosphorylation proteins, during thermogenesis induction in the visceral and epididymal fat in mice with HFD-induced obesity. Notably, the physiological and metabolic changes were more significant after treatment with potential probiotic culture-supernatants than those with the bacterial pellet. Finally, gene knockdown and co-treatment with inhibitor-mediated mechanistic analyses showed that both DS0908 and DS0950 exerted anti-obesity-related effects via the PKA/p38 MAPK signaling activation in C3H10T1/2 MSCs. Our observations suggest that DS0908 and DS0950 could potentially alleviate obesity as dietary supplements.

Effects of Substance P on the Release of Cytokines from Immune Cell Lines (면역세포의 cytokine 유리에 미치는 substance P의 영향)

  • Lee, Jin-Yong;Kim, Soo-Ah;Seo, Seok-Ran;Kim, Hyong-Seop
    • Journal of Periodontal and Implant Science
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    • v.27 no.2
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    • pp.425-441
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    • 1997
  • The neuropeptide substance P(SP) has been implicated in the mediation of inflammation and immune-mediated disease such as arthritis. Recently, it was reported that SP was markedly increased around the blood vessels in inflamed gingiva as well as in close association with the inflammatory cell infiltrate. These results support that SP may contribute to the pathophysiology of neuronal inflammation in human periodontal tissues. SP may regulate inflammatory/immune responses by stimulating the proliferation of human T cells, differentiation and antibody-secreting potential of B cells, macrophage respiratory burst, connective tissue proliferation, and the secretion of cytokines from monocytes and T cells. Here, I studied potential role of SP as a costimulatory chemical signal in inflammatory/immune responses, by determining the released proinflammatory cytokines such as $MIP-1{\alpha}$, $IL-1{\beta}$, and IL-6 from culture supernatants of homogeneous immune cell lines. Serum free cell supernatants were concentrated with TCA precipitation, fractionated with SDS-PAGE, and subjected into western blot analysis. Among 15 cell lines tested, macrophage/monocyte cell line RAW264.7 and WRl9m.1 showed the highest level of induction of $MIP-1{\alpha}$ when stimulated with LPS. Discrete IL-6 bands with multiple forms of molecular mass were detected from supernatants of B cell lines A20(32kDa), Daudi(32, 35kDa), and SKW6.4(29kDa), which were expressed constitutively. $IL-1{\beta}$ could not be detected by the method of western blot analysis from supernatants of all cell lines tested except RAW264.7, WRl9m.1, and erythroid cell line K562 which showed the least amount of $IL-{\beta}$ secretion. SP $10^{-9}M$ with suboptimal dose of LPS treatment showed synergistic induction of $MIP-1{\alpha}$ release from RAW264.7 or WR19m.1, and also IL-6 release from A20, but this synergism is not the case in costimulation of RAW264.7 or WRl9m.1 with SP $10^{-9}M$ and TPA. Although treatment of T cell line CTLL-R8 with SP $10^{-7}M$ or PHA+TPA induced modest level of $MIP-1{\alpha}$ secretion, synergism was not observed when they are applied together. These findings all together suggest the possibility of a regulatory role of SP in inflammatory/immune reaction through differential modulation of bioactivities of other chemical cosignals.

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Studies on the Functional Properties of Lactic Acid Bacteria Isolated from Home-made Yogurt and Commercial Yogurt (Home-made 요구르트와 시판 중인 요구르트에서 분리한 젖산균의 기능적 특성 조사)

  • Choi, Moon-Sup;Yun, Hyun-Myoung;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.8-14
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    • 2014
  • The objective of this work is to investigate and compare several functional properties of lactic acid bacteria (LAB), Lactobacillus casei SK-7 isolated from home-made yogurt and Lactobacillus bulgaricus YK-11 from commercial yogurt. Initially, physiological and biochemical properties of SK-7 and YK-11 were characterized. Phylogenetic analysis using 16S rRNA sequencing were performed to identify the strains, and the strain could be assigned to Lactobacillus casei and Lactobacillus bulgaricus, designated as L. casei SK-7 and L. bulgaricus YK-11. Phylogenetic tree of SK-7 and YK-11 was plotted based on 16S rRNA sequence comparisons. Production of lactic acid and organic acid, and pH changes in the cultures of SK-7 and YK-11 were monitored during 72 h. During the incubation period, several functional properties of L. casei SK-7 and L. bulgaricus YK-11 were examined. L. casei SK-7 and L. bulgaricus YK-11 cultures eliminated 93.9% and 88.2% of nitrite, respectively. Antioxidant activity of cultural supernatants of SK-7 and YK-11 were 62.6%, 54.9%, and activity of ${\beta}$-galactosidase were 14.9 units/mg and 13.1 units/mg, respectively. The antimicrobial activities were examined with 20-fold concentrated culture supernatants from the cultures of SK-7 and YK-11. The activities of SK-7 supernatants were clearly observed against all microorganisms in this work, whereas no activities were observed in YK-11 supernatants. Although it might be conducted additional functional research, functional properties of LAB isolated from home-made yogurt have been shown to be better than those of commercial yogurt in this work.

Relative Level of Sucrose Metabilizing Enzymes in Oral Streptococci (구강 Streptococci가 가진 Sucrose 대사 효소의 활성도의 비교)

  • 최선진
    • Korean Journal of Microbiology
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    • v.19 no.3
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    • pp.137-141
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    • 1981
  • Occurrence and distribution of sucrose metabolizing enzymes in oral streptococci had been studied. In these studies, the carbohydrate component of the culture medium had been glucose. I have extended these studies by analyzing bacterial culture supernatants for the relative content of hexosyltransferases, namely glucosyl and fructosyltransferase. As a carbohydrate, fructose was used. The growth measured for nine oral streptococci (Strptococcus mutans strains BHT, ING, AHT, 6715, LM-7, and SL-1 ; Streptococcus sanguis 903, 9811, and M-5) varied. The level of glucosyltansferase activity also varied among S. mutans strains, and its level in S. sanguis was relatively low. Fructosyltansferase activity of the various strains fluctuated more than of glucosyltransferase. S.mutans strain LM-7 had significantly higher level of both enzymes. As a whole, fructose-grown cultures had generally an agreeable trend of enzyme activity to those from glucose-grown cultures.

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