• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1251-1258
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• 2010

During the nitrogen-fixing process, ammonia ($NH_3$) is incorporated into glutamate to yield glutamine and is generally not secreted. However, in this study, $NH_3$-excreting strains of nitrogen-fixing Paenibacillus were isolated from soil. The ammonium production by the Paenibacillus strains was assayed in different experiments (dry biomass, wet biomass, cell-free extract, and cell-free extract adsorbed on nano $TiO_2$ particles) inside an innovative bioreactor containing capsules of $N_2$ and $H_2$. In addition, the effects of different $N_2$ and $H_2$ treatments on the formation of $NH_3$ were assayed. The results showed that the dry biomass of the strains produced the most $NH_3$. The dry biomass of the Paenibacillus strain E produced the most $NH_3$ at 1.50, 0.34, and 0.27 ${\mu}M$ $NH_3$/mg biomass/h in the presence of $N_2$ + $H_2$, $N_2$, and $H_2$, respectively, indicating that a combined effluent of $N_2$ and $H_2$ was vital for $NH_3$ production. Notwithstanding, a cell-free extract (CFE) adsorbed on nano $TiO_2$ particles produced the most $NH_3$ and preserved the enzyme activities for a longer period of time, where the $NH_3$ production was 2.45 ${\mu}M$/mg CFE/h over 17 h. Therefore, the present study provides a new, simple, and inexpensive method of $NH_3$ production.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.4
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• pp.203-210
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• 2011

Cilostazol is a selective inhibitor of phosphodiesterase 3 that increases intracellular cAMP levels and activates protein kinase A, thereby inhibiting vascular smooth muscle cell (VSMC) proliferation. We investigated whether AMP-activated protein kinase (AMPK) activation induced by heme oxygenase-1 (HO-1) is a mediator of the beneficial effects of cilostazol and whether cilostazol may prevent cell proliferation and reactive oxygen species (ROS) production by activating AMPK in VSMC. In the present study, we investigated VSMC with various concentrations of cilostazol. Treatment with cilostazol increased HO-1 expression and phosphorylation of AMPK in a dose- and time-dependent manner. Cilostazol also significantly decreased platelet-derived growth factor (PDGF)-induced VSMC proliferation and ROS production by activating AMPK induced by HO-1. Pharmacological and genetic inhibition of HO-1 and AMPK blocked the cilostazol-induced inhibition of cell proliferation and ROS production.These data suggest that cilostazol-induced HO-1 expression and AMPK activation might attenuate PDGF-induced VSMC proliferation and ROS production.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.23 no.2
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• pp.69-80
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• 2010

Objective : The purpose of this study was to compare anti-Inflammation and anti-oxidation of Jeondo-San(JDS) extracted with two kinds of solvents, ethanol and water. Methods : Two kinds of JDS extractions were prepared 20, 50, $100\;{\mu}g/mg$. The Cytotoxicity was measured by MTT assay in Raw 264.7 cell. The anti-inflammation effects were measured by inhibitory efficacy on $PGE_2$, NO, TNF-$\alpha$, COX-2 and iNOS in Raw 264.7 cell. The anti-oxidation effects were measured by ROS inhibitory efficacy, intracellular GSH synthesis and DPPH Radical scavenging in HaCaT cell. Results : 1. All of JDS extraction groups had no cytotoxicity in Raw 264.7 cell. 2. All of JDS extraction groups showed significantly inhibitory effect on production of $PGE_2$. Inhibitory efficacy increased in accodance with concentration. 3. All of JDS extraction groups showed significantly inhibitory effect on production of NO. Inhibitory efficacy increased in accodance with concentration. 4. All of JDS extraction groups did not show significantly inhibitory effect on production of TNF-$\alpha$. 5. $100\;{\mu}g/ml$ JDS extracted with ethanol and $50\;{\mu}g/ml$, $100\;{\mu}g/ml$ JDS extracted with water showed inhibitory effect on iNOS expression. 6. All of JDS extraction groups showed significantly inhibitory effect on production of ROS. Inhibitory efficacy increased in accodance with concentration. Ethanol extractions were better than water extractions. 7. $100\;{\mu}g/ml$ JDS extracted with ethanol only produced GSH of $32{\pm}5.2%$. 8. All of JDS extraction groups showed significantly scavenging effect of DPPH radicals. Inhibitory efficacy increased in accodance with concentration. Ethanol extractions were better than water extractions. Conclusion : Two kinds of JDS extractions have not cytotoxicity and inhibit production of NO. JDS extracted with water was effective in anti-inflammation, JDS extracted with ethanol was effective in anti-oxidation.

• YAKHAK HOEJI
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• v.38 no.6
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• pp.806-813
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• 1994

These experiments were conducted to investigate the effects of Cervi cornu extract on lymphocyte blastogenesis in spleen, thymus, lymph node, born marrow cells of Balb/c mouse, haemagglutination reaction against sheep red blood cell (SRBC), plaque forming cell (PFC) assay against SRBC and IL-2 production. Lymphocyte blastogenesis was determined by $[^3H]-thymidine$ incorporation. According to the lymphcoyte blastogenesis test on the immune cell. Ceriv cornu extrat was showed a potent mitogenic activity on the spleen and lymph node cells, but had mild mitogenic activity on the thymus and born marrow cells. Mitogenic active component of Crevi cornu extract was identified to be materials where molecular weights are higher than 5,000 by membrane filteration method. Cervi cornu extrat was shown to increase mitogenic effect on the lipopolysaccharide (LPS)-stimulated spleen cells significantly, but decrease mitogenic effect on the Con A stimulated spleen cell at the concentration 0.3%, 1% and 3%. Ceriv cornu extract didn't show to be haemagglutination reaction and showed to inhibit the Con A-induced haemagglutination reaction against SREC. Result of SRBC-PEC test. Ceriv cornu extract significantly increase the number of PEC at the concentration of 0.1% and 1%. When IL-2 or IL-4 production was determined by proliferation of CTLL-2 cells. Ceriv cornu extract was not shown to stimulate the production of IL-2. From the above results, it is shown that Ceriv cornu extract increased antibody production by B cells, but nor IL-2 production by helper T cells.

• Journal of the Korean Society of Food Science and Nutrition
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• v.28 no.5
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• pp.997-1002
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• 1999

The aim of the present study was to investigate effects of food preservative addition and changes in composition of herb extract product on the growth of spoilage yeast, Zygosaccharomyces sp. Herbs such as Panax ginseng, Cinnamomum cassia, Lycium chinense, Zyzyphus juiuba and Jingiber officinale were altogether put into water and essence was extracted at 80oC, and then the extract was concentrated at 75oC. The herb extract product was made by adding vitamins, amino acids and honey to the concentrated herb extract. The amount of gas produced from the herb extract product was increased as inoculated cell number increased but decreased as Brix concentration increased. Gases were produced in small amount when incubation was made at 4oC but large amounts of gases were produced at 25 or 40oC of incubation. The gas production and growth of Zygosaccharomyces sp. were measured after browning reaction was induced by heating at 85oC for 12 hours. It appeared that heating treatment did not induce any significant change in the gas production and growth of the cell. The effects of addition of various sugar to the herb extract produce were also invesigated. Amounts of gas production were in the order of glucose>sucrose>oligosaccharide>stevioside. The viable cell count was measured as 6.0$\times$107 CFU/g when glucose was added to the herb extract product. The viable cell counts were 5.0$\times$106, 3.0$\times$103, and 3.0$\times$102 CFU/g in sucrose, oligosaccharide and stevioside added herb extract product, respectively. The amount of gas production from the herb extract product was remarkably reduced by addition of such food preservatives as sodium benzoate and DF 100. TLC(thin layer chromatography) chromatogram of the herb extract showed stability of the herb extract in the above treatments.

• IMMUNE NETWORK
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• v.14 no.2
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• pp.89-99
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• 2014

Graft-versus-host disease (GVHD) is a fatal complication that occurs after allogeneic hematopoietic stem cell transplantation. To understand the dynamics of CD4 and CD8 T cell production of IFN-${\gamma}$ and IL-17 during GVHD progression, we established a GVHD model by transplanting T cell-depleted bone marrow (TCD-BM) and purified T cells from B6 mice into irradiated BALB.B, creating an MHC-matched but minor histocompatibility (H) antigen-mismatched transplantation (B6 ${\rightarrow}$ BALB.B GVHD). Transplantation-induced GVHD was confirmed by the presence of the appropriate compositional changes in the T cell compartments and innate immune cells in the blood and the systemic secretion of inflammatory cytokines. Using this B6 ${\rightarrow}$ BALB.B GVHD model, we showed that the production of IFN-${\gamma}$ and IL-17 by CD4 T cells preceded that by CD8 T cells in the spleen, mesenteric lymph node, liver, and lung in the BALB.B GVHD host, and Th1 differentiation predated Th17 differentiation in all organs during GVHD progression. Such changes in cytokine production were based on changes in cytokine gene expression by the T cells at different time points during GVHD development. These results demonstrate that both IFN-${\gamma}$ and IL-17 are produced by CD4 and CD8 T cells but with different kinetics during GVHD progression.

• Microbiology and Biotechnology Letters
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• v.16 no.4
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• pp.327-333
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• 1988

For optimal production of recombinant human interleukin-2 (IL-2) in E. coli the effect of fermentation conditions on cell growth, IL-2 production, and stability of recombinant cells were investigated. Among the complex nutrients tested in this work, yeast extract, peptone and corn steep liquor were found to be effective for recombinant cell growth. The recombinant cells were maintained stably under repression condition (3$0^{\circ}C$), but the stability of recombinant cells were drastically reduced upon induction of IL-2 expression (42$^{\circ}C$) even under the selection pressure. Addition of antibiotics to the culture medium resulted in the cell growth inhibition without significant improvement in recombinant stability. When the expression of IL-2 gene was induced at different growth phases, highest IL-2 production was achieved by the induction of IL-2 at the middle-exponential growth phase. It was found that the production of IL-2 significantly inhibited the cell growth and the ex-pression of other genes in the plasmid.

• Journal of Microbiology and Biotechnology
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• v.27 no.11
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• pp.1952-1960
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• 2017

Recently, soybean isoflavone aglycones (i.e., daidzein and genistein) and ${\gamma}-aminobutyric$ acid (GABA) have begun to receive considerable consumer attention owing to their potential as nutraceuticals. To produce these ingredients, multiple microorganisms and their enzymes are commonly used for catalysis in the nutraceutical industry. In this work, we introduce a novel fermentation process that uses whole-cell biocatalysis to accelerate GABA and isoflavone aglycone production in doenjang (a traditional Korean soybean paste). Microbial enzymes transform soybean isoflavone glycosides (i.e., daidzin and genistin) and monosodium glutamate into soybean isoflavone aglycones and GABA. Lactobacillus brevis GABA 100 and Aspergillus oryzae KACC 40250 significantly reduced the production time with the aid of a protease. The resulting levels of GABA and daidzein were higher, and genistein production resembled the levels in traditional doenjang fermented for over a year. Concentrations of GABA, daidzein, and genistein were measured as 7,162, 60, and $59{\mu}g/g$, respectively on the seventh day of fermentation. Our results demonstrate that the administration of whole-cell L. brevis GABA 100 and A. oryzae KACC 40250 paired with a protease treatment is an effective method to accelerate GABA, daidzein, and genistein production in doenjang.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.257-262
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• 1989

The induction of calli from tissues of a grape, Vitis hybrid, and their suspension cultures were performed and various factors were investigated on cell growth and anthocyanin production. It was shown that light intensity and inorganic nitrogen concentration played an important role on anthocyanin production.1:he contents of anthocyanin produced under 10,000 Iux light irradiation were about twice as much as under the dark. The reduction of inorganic nitrogen concentration of MS medium to one to twenty brought about the increase of approximately five to six-fold in total anthocyanin or sixteenfold in anthocyanin content per dry cell weight and addition of nitrate only as inorganic nitrogen source was shown to be the best for anthocyanin production. Miller medium and Gamborg medium were suitable for the anthocyanin production, as well as high concentrations of Co$^{2+}$ and Fe$^{2+}$. And high yield of 40mg anthocyanins per 200m1 flask was obtained by two stage culture using MS medium for the first stage and the modified MS medium for the second stage.

• Journal of Acupuncture Research
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• v.35 no.4
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• pp.182-186
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• 2018

Background: The purpose of this study was to investigate the effects of wild ginseng pharmacopuncture on melanin production in B16/F10 murine melanoma cells. Methods: To determine the effect of wild ginseng pharmacopuncture solution on B16/F10 cells, cytotoxicity was evaluated using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyl-tetrazolium bromide (MTT) method. To observe B16/F10 cell growth, death, and morphological changes, Trypan blue solution was used. The Hosoi method was used to investigate the effect of wild ginseng pharmacopuncture solution on melanin production. The Martinez-Esparza method was used to investigate the effect of wild ginseng pharmacopuncture solution on tyrosinase activity. To determine the pathway involved in the melanogenesis in cells exposed to wild ginseng pharmacopuncture solution, a cell-free tyrosinase was used. Results: Following treatment with $200{\mu}L$ of wild ginseng solution, the cell survival rate was $76.32{\pm}2.45%$ which significantly decreased with higher concentrations (${\mu}L$) of wild ginseng (up to $200{\mu}L$). When $100{\mu}L$ of wild ginseng was used, the cell survival rate was $89.95{\pm}2.07%$. No morphological changes or abnormalities were observed in the B16/F10 murine melanoma cells as observed in the Trypan blue test. Melanin production was significantly reduced to $72.17{\pm}3.74%$ at $100{\mu}L$. Using $100{\mu}L$ of wild ginseng solution, tyrosinase activity was significantly decreased to $80.15{\pm}1.05%$. Wild ginseng pharmacopuncture solution reduced melanin production both directly and indirectly. Conclusion: This study suggests that wild ginseng pharmacopuncture solution may be effective in inhibiting melanin production. Further studies are needed to determine safe and effective clinical applications.