• Journal of Veterinary Clinics
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• v.31 no.6
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• pp.469-476
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• 2014

The aims of this study were to explore the effects of conjugated linoleic acid (CLA) on reactive oxygen species (ROS) production in lipopolysaccharide (LPS)-naïve and LPS-stimulated RAW 264.7 macrophages and to examine whether these effects affect the regulation of tumor necrosis factor-alpha (TNF-${\alpha}$) production, and nuclear factor-kappa B (NF-${\kappa}B$) and peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) activation. Trans-10, cis-12(t10c12)-CLA increased the production of ROS, as well as TNF-${\alpha}$ in LPS-naïve RAW 264.7 cells. The CLA-induced TNF-${\alpha}$ production was suppressed by treatment of diphenyleneiodonium chloride (DPI), a NADPH oxidase inhibitor. In addition, CLA enhanced the activities of NF-${\kappa}B$ and $PPAR{\gamma}$ in LPS-naïve RAW 264.7 cells, and this effect was abolished with DPI treatment. LPS treatment increased ROS production, whereas CLA reduced LPS-induced ROS production. LPS increased both TNF-${\alpha}$ production and NF-${\kappa}B$ activity, whereas t10c12-CLA reduced TNF-${\alpha}$ production and NF-${\kappa}B$ activity in LPS-stimulated RAW 264.7 cells. DPI treatment suppressed LPS-induced ROS production and NF-${\kappa}B$ activity. Moreover, DPI enhanced the inhibitory effects of t10c12-CLA on TNF-${\alpha}$ production and NF-${\kappa}B$ activation in LPS-stimulated RAW 264.7 cells. However, neither t10c12-CLA nor DPI affected $PPAR{\gamma}$ activity in LPS-stimulated RAW 264.7 cells. Taken together, these data indicate that t10c12-CLA induces TNF-${\alpha}$ production by increasing ROS production in LPS-naïve RAW 264.7 cells, which is mediated by the enhancement of NF-${\kappa}B$ activity via $PPAR{\gamma}$ activation. By contrast, t10c12-CLA suppresses TNF-${\alpha}$ production by inhibiting ROS production and NF-${\kappa}B$ activation via a $PPAR{\gamma}$-independent pathway in LPS-stimulated RAW 264.7 cells. These results suggest that t10c12-CLA can modulate TNF-${\alpha}$ production and NF-${\kappa}B$ activation through formation of ROS in RAW 264.7 macrophages.

• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1061-1070
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• 2019

In the present study, the optimization of poly(${\gamma}$-glutamic acid) (${\gamma}$-PGA) production by Bacillus sp. FBL-2 was studied using a statistical approach. One-factor-at-a-time method was used to investigate the effect of carbon sources and nitrogen sources on ${\gamma}$-PGA production and was utilized to select the most significant nutrients affecting the yield of ${\gamma}$-PGA. After identifying effective nutrients, response surface methodology with central composite design (CCD) was used to obtain a mathematical model to identify the optimum concentrations of the key nutrients (sucrose, $\text\tiny{L}$-glutamic acid, yeast extract, and citric acid) for improvement of ${\gamma}$-PGA production. The optimum amount of significant medium components appeared to be sucrose 51.73 g/l, $\text\tiny{L}$-glutamic acid 105.30 g/l, yeast extract 13.25 g/l, and citric acid 10.04 g/l. The optimized medium was validated experimentally, and ${\gamma}$-PGA production increased significantly from 3.59 g/l (0.33 g/l/h) to 44.04 g/l (3.67 g/l/h) when strain FBL-2 was cultivated under the optimal medium developed by the statistical approach, as compared to non-optimized medium.

• The Korean Journal of Food And Nutrition
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• v.25 no.2
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• pp.362-367
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• 2012

Corn has been used for a long time as a traditional remedy, as well as a food source. We previously reported that in vitro supplementation of corn water extracts enhanced the proliferation of splenocytes, compared to the control group. In this study, we examined the immunomodulative effect of a water extract of corn. Seven to eight weeks old mice(Balb/c) were fed an ad libitum chow diet, and were orally administrated a water extract of corn every other day, for four weeks, at two different concentrations(50 and 500 mg/kg B.W). Cytokine production(IL-2, IL-10 and IFN-${\gamma}$) by macrophages stimulated with LPS or not stimulated with LPS was detected by ELISA assay using the cytokine kit. In an ex vivo study, the cytokines IL-2, IL-10 and IFN-${\gamma}$ were detected at 500 mg/kg b.w. supplementation group with LPS stimulation in all cases. Also, the ratio of IFN-${\gamma}$ to IL-10 was in the range of 0~3 with mitogen stimulation, such as con A and LPS. In conclusion, this study suggests that in mice, corn extracts may enhance immune function by regulating the cytokine production(IL-2, IL-10 and IFN-${\gamma}$) of the activated macrophages.

• Parasites, Hosts and Diseases
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• v.42 no.4
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• pp.175-183
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• 2004

To evaluate the role of nitric oxide (NO) in $IFN-{\gamma}$ production and apoptosis of splenocytes in genetically different strains of mice with toxoplasmosis, BALB/c (a toxoplasmosis resistant strain) and C57BL/6 (a toxoplasmosis susceptible strain) mice were infected with Toxoplasma gondii cysts orally and subsequently injected intraperitoneally with aminoguanidine, an iNOS inhibitor (AG; 35 mg/kg per mouse daily for 14 days). When BALB/c or C57BL/6 mice were infected with T. gondii without AG treatment, number of brain cysts, NO and IFN-y production by splenocytes, and percentages of apoptotic splenocytes were increased compared to uninfected control mice without AG treatment. AG treatment increased the number of brain cysts, and reduced NO and $IFN-{\gamma}$ production in T. gondii-infected C57BL/6 mice. In contrast, in T. gondii-infected BABL/c mice, the number of brain cysts, and NO and $IFN-{\gamma}$ production of splenocytes was not altered by treatment with AG. However, the percentages of apoptotic splenocytes in T. gondii-infected BALB/c or C57BL/6 mice were not affected by AG treatment. These results suggest that NO modulates $IFN-{\gamma}$ production in T. gondii-infected C57BL/6 mice, and that NO is involved in mediating a protective response in toxoplasmosis susceptible, but not resistant, mice strain during acute infection.

• Journal of Acupuncture Research
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• v.27 no.4
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• pp.223-231
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• 2010

Objectives : The objective of this study is to study the effects of Euonymi Lignum Suberatatum pharmacopuncture solution on NO and IL-6 production in macrophage. Methods : At first, the RAW 264.7 macrophage was subclutured. In order to evaluate cytotoxicity, MTT assay performed. Then, the cell was induced by LPS, INF-$\gamma$ and Experimental groups were divided into five(Normal, Control, Euonymi Lignum Suberatatum 100, 200, $300{\mu}g/m{\ell}$). Then Euonymi Lignum Suberatatum pharmacopuncture solution was put into cell. We measured IL-6, iNOS, NO. Results : The cytotoxic effect of Euonymi Lignum Suberatatum pharmacopuncture solution in RAW 264.7 macrophage was not appeared. $300{\mu}g/m\ell$ Euonymi Lignum Suberatatum pharmacopuncture solution inhibited IL-6 production in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages significantly. Euonymi Lignum Suberatatum pharmacopuncture solution inhibited iNOS revelation in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages. All group of Euonymi Lignum Suberatatum pharmacopuncture solution inhibited NO production in LPS, INF-$\gamma$-stimulated RAW 264.7 macrophages significantly. Conclusions : Our study demonstrated that Euonymi Lignum Suberatatum pharmacopuncture solution had an inhibition effect on NO production, iNOS revelation, IL-6 production. So Euonymi Lignum Suberatatum pharmaco puncture solution may have an Anti-inflammation effect.

• The Journal of Internal Korean Medicine
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• v.20 no.1
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• pp.73-82
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• 1999

Nitric oxide (NO) is now recognized as a mediator of several biological and immunological functions, but unlike classical neurotransmitters. NO simply diffuse of the postsynaptic cells and around affecting cells. Taraxacum officinale (Compositae) has been used for maintenance of vitality, and they still occupy an important place in the traditional Korean medicine. We have examined that the effect of Taraxacum officinale water extract on NO synthesis in microglial cells of murine's brain, using the Griess method. And this study was evident that Taraxacum officinale did not induce NO production without recombinant interferon gamma ($rIFN-{\gamma}$), whereas Taraxacum officinale (10-1000 g/ml) with $rIFN-{\gamma}$ effectively produced NO in microglial cells of brain. As result. NO production in microglial cells increased most significantly in dose of 100 g/ml of the Taraxacum officinale and the production of NO was dependent on the dose of Taraxacum officinale, NG-monomethyl-L-arginine, competitive inhibitor of NO synthase, reduced the NO production by Taraxacum officinale stimulation with $rIFN-{\gamma}$ in microglial cells of murine. The effect of Taraxacum officinale was mainly dependent on Taraxacum officinale-induced tumor necrosis factor- secretion. Conclusively, this study suggested that Taraxacum officinale stimulate NO production at microglial cells in brain, which may be an important factor for mediating immune and neuroendocrinologic regulation in nervous system.

• Preventive Nutrition and Food Science
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• v.4 no.3
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• pp.175-179
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• 1999

To develop a production method for modified starches with less pollution, pH adjustment and gamma irradiation were applied to commerical corn starch. Blue values were significantly decreased , while alkali number, optical transmittance and solubility markedly increased when gamma irradiation was applied to pH 2 adjusted corn starch. Water binding capacity and swelling power at pH 5 were the highest among the samples. Gelatinization viscosity was considerably affected by gamma irradiation and pH of the starch. Gamma irradiation of pH 2 adjusted starch showed the lowest peak viscosity and the best cooling stability among the tested samples . Therefore, the production of modified starch with low viscosity as well as with sufficient viscosity stability seems feasible by controlling the pH of the starch and gamma irrdiation.

• Korean Journal of Medicinal Crop Science
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• v.15 no.5
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• pp.324-328
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• 2007

To search for immunoactive natural products exerting anti-inflammatory activity, we have evaluated the effects of the ethanol extracts of Rubus coreanus Miq. (ERC) on lipopolysaccharide-induced nitric oxide (NO), tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$, and $Interferon-{\gamma}\;(IFN-{\gamma})$ production by RAW 264.7 macrophage cell line. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased $IFN-{\gamma}\;and\;TNF-{\alpha}$ production. Consistent with these results, the protein level of inducible Nitric Oxide Synthase (iNOS) and cyclooxygenase-2 (COX-2) was inhibited by ethanol extracts of ERC in a dose-dependent manner. These results suggest that ERC may exert anti-inflammatory and analgesic effects possibly by suppressing the inducible NO synthase and COX-2 expressions.

• Microbiology and Biotechnology Letters
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• v.22 no.4
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• pp.389-394
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• 1994

8 X 10$^{6}$(viable cells/ml) of maximum cell density and 9000(IU/ml) of $\gamma$-IFN production were obtained at 55(ml/hr) of a perfusion rate by cultivating HSF cells using a moving membrane aeration bioreactor. This system proves to be an efficient culture process by maintaning 90% of viable cells during the whole cultivation periods. The metabolic molar quotient of glucose to lactate was 0.81 for overall ranges of glucose consumed while the evolution of ammonia was not linearly related to the consumption of glutamine. Low molar conversion ratio was observed in low consumptions of glutamine and high molar conversion ratio in high comsumptions. It also shows that the glutamolysis plays important role in the steady state conditions by evolving larger quantities of ammonia than lactate. At the above of 50 rpm, which is the optimum agitation speed for this bioreactor, the cell growth was severely affected while the IFN production was less decrea- sed, maintaing 1.5 X 10$^{-3}$(IU/cell/day) specific IFN production rate. The cumulatvie $\gamma$-IFN production was 7.2 X 10$^{8}$(IU) for 70 days of the cultivation, which yields 1 X 10$^{7}$ (IU/day) of IFN production rate. Therefore, a commercial production of $\gamma$-IFN by this culture process can be achievable by maintaining the above IFN productivity in a scaled-up culture system.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1995.04a
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• pp.78-78
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• 1995

Increasing evidence indicates that the production of nitric oxide (NO) by inducible NO synthase (NOS) is tightely regulated. Transforming growth factor-${\beta}$ (TGF-${\beta}$) is a homodimeric protein secreted during macrophage activation, but several lines of evidence suggest that TGF-${\beta}$ is selectively suppressive for macrophage NO production. We therefore reasoned that a strategy employing oligodeoxynucleotides(ODNs) complemently to TGF-${\beta}$ mRNA (antisense ODNs) might increase NO production in IFN-${\gamma}$-treated murine peritoneal macrophages. To evaluate this concept, we tested the effects of antisense ODNs targeted to TGF-${\beta}$ mRNA (25-mer ODNs complemently to TGF-${\beta}$mRNA sequences) by introducing it into the medium of cultured macrophages. Phosphorothiolation of ODNs were employed to retard their degradation. Antisense ODNs had no effect on NO production by itself, whereas IFN-${\gamma}$ alone had modest effect. When antisense ODNs were used in combination with IFN-${\gamma}$, there was a marked cooperative induction of NO production, These effects of antisense ODNs were associated with decreased TGF-${\beta}$ expression in activated macrophages. ODNs with the same nucleotides but a scrambled sequence had no effect. Adding anti-TGF-${\beta}$ antibodies to the IFN-${\gamma}$-treated macrophages mimicked the positive effect of antisense ODNs on NO production. In addition, the effects of either antisense ODNs or anti-TGF-${\beta}$ antibodies were blocked by adding TGF-${\beta}$ in cultured macrophages. These results indicate that the generation of TGF-${\beta}$ by activated macrophages provides a self-regulating mechanism by which the temporal and perhaps spatial production of NO, a reactive and potentially toxic mediator, can be finely regulated.