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Potential immune-modulatory effects of wheat phytase on the performance of a mouse macrophage cell line, Raw 264.7, exposed to long-chain inorganic polyphosphate

  • An, Jeongmin;Cho, Jaiesoon
    • Animal Bioscience
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    • v.34 no.3_spc
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    • pp.463-470
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    • 2021
  • Objective: This experiment was conducted to find out the immunological effects of wheat phytase when long-chain inorganic polyphosphate (polyP) treated with wheat phytase was added to a macrophage cell line, Raw 264.7, when compared to intact long-chain polyP. Methods: Nitric oxide (NO) production of Raw 264.7 cells exposed to P700, a long-chain polyP with an average of 1,150 phosphate residues, treated with or without wheat phytase, was measured by Griess method. Phagocytosis assay of P700 treated with or without phytase in Raw 264.7 cells was investigated using neutral red uptake. The secretion of tumor necrosis factor α (TNF-α) by Raw 264.7 cells with wheat phytase-treated P700 compared to intact P700 was observed by using Mouse TNF-α enzyme-linked immunosorbent assay kit. Results: P700 treated with wheat phytase effectively increased NO production of Raw 264.7 cells by 172% when compared with intact P700 at 12 h exposure. At 5 mM of P700 concentration, wheat phytase promoted NO production of macrophages most strongly. P700, treated with wheat phytase, stimulated phagocytosis in macrophages at 12 h exposure by about 1.7-fold compared to intact P700. In addition, P700 treated with wheat phytase effectively increased in vitro phagocytic activity of Raw 264.7 cells at a concentration above 5 mM when compared to intact P700. P700 dephosphorylated by wheat phytase increased the release of TNF-α from Raw 264.7 cells by 143% over that from intact P700 after 6 h exposure. At the concentration of 50 μM P700, wheat phytase increased the secretion of cytokine, TNF-α, by 124% over that from intact P700. Conclusion: In animal husbandry, wheat phytase can mitigate the long-chain polyP causing damage by improving the immune capabilities of macrophages in the host. Thus, wheat phytase has potential as an immunological modulator and future feed additive for regulating immune responses caused by inflammation induced by long-chain polyP from bacterial infection.

Survival Behavior of Escherichia coli O157:H7 during over 60-Day Aging of Camembert Cheeses Manufactured from Unpasteurized Raw Milk

  • Chon, Jung-Whan;Seo, Kun-Ho;Lim, Jong-Soo;Youn, Hye-Young;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • v.40 no.4
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    • pp.151-162
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    • 2022
  • Although many developed countries (USA, Canada, and several EU countries) allow raw milk cheese to be aged more than 60 days, these countries have strict standards for the aging conditions, such as temperature, of raw milk cheese. Spiking experiments were conducted with Camembert cheese made from raw milk, to assess the microbiological safety of raw milk cheese aged for more than 60 days. We spiked Escherichia coli O157:H7 into raw milk with different inoculation levels (high, medium, and low). Camembert cheese was prepared from the inoculated raw milk, then aged in an incubator for up to 9 weeks (63 days). There were no significant differences in pH and water activity (aW) between uninoculated cheese and cheese samples inoculated with E. coli O157:H7 (p<0.05). The pH and aWof the Camembert cheese decreased throughout the storage period. In conclusion, E. coli O157:H7 did not affect the pH and aW of the cheese samples. Cell counts were conducted every week using the agar-plating method. Inoculated cells were completely eliminated, especially in Camembert cheese, after 60 days, and the reduction rate of cells was much faster in Camembert cheese.

Anti-inflammatory Effects of Actinidia Polygama Ethanol Extract in through the Regulated NF-κ B and MAPKs Activation in LPS Stimulated RAW 264.7 Cells (RAW 264.7 세포에 대한 NF-κ B와 MAPK 활성 억제를 통한 개다래 열매 에탄올 추출물의 항염증 효과)

  • Chung-Mu Park;Hyun-Seo Yoon
    • Journal of The Korean Society of Integrative Medicine
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    • v.11 no.2
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    • pp.119-128
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    • 2023
  • Purpose : The fruit of Actinidia polygama has been used in oriental medicine for the treatment of gout, rheumatoid arthritis, and inflammation. Though A. polygama exhibited anti-inflammatory activity in RAW 264.7 cells and carrageenan-induced rat paw edema, the exact mechanism for anti-inflammation was not evaluated yet. In this study, the anti-inflammatory mechanisms of A. polygama ethanol extract (APEE) in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Methods : WST-1 assay was applied to analyze the cytotoxic effect of APEE in RAW 264.7 cells. The productions of nitric oxide (NO) and prostaglandin (PG) E2 were analyzed by the Griess reaction and enzyme immunoassay (EIA) assay, respectively. In addition, protein expressions for inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 were measured by Western blot analysis. The activated status of an inflammatory transcription factor, NF-κ B, and its upstream signaling molecules, mitogen-activated protein kinases (MAPKs), was also evaluated by Western blot analysis. Results : As a result, APEE treatment did not exhibit any cytotoxicity until the concentration of 200 ㎍/㎖. APEE treatment significantly inhibited NO and PGE2 productions as well as their enzymes, iNOS and COX-2 in a dose-dependent manner. The inflammatory transcription factor, NF-κ B, was also attenuated by APEE treatment. In addition, the phosphorylated status of MAPKs such as extracellular regulated kinase (ERK), c-jun NH2 kinase (JNK), and p38, were significantly diminished by APEE treatment in LPS stimulated RAW 264.7 cells. Conclusion : Consequently, APEE treatment significantly attenuated the production of inflammatory mediators and their enzyme expressions in LPS-stimulated RAW 264.7 cells. The inflammatory transcription factor, NF-κ B, and upstream signaling molecules, MAPKs, were also significantly attenuated by APEE treatment in LPS-activated RAW 264.7 cells. These results indicate that APEE might be a candidate to be utilized as a promising candidate for the treatment of inflammatory disorders.

Effects of baicalein on hydrogen peroxide productions in mouse macrophages stimulated by lipopolysaccharide and peptidoglycan (지질다당체와 펩티도글라이칸 공동 자극으로 유발되는 대식세포의 하이드로겐 퍼록사이드 생성증가에 미치는 바이칼레인의 작용 고찰)

  • Wansu Park
    • The Korea Journal of Herbology
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    • v.38 no.6
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    • pp.45-52
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    • 2023
  • Objectives : Effects of baicalein (BA) on oxidative stress in RAW 264.7 mouse macrophages stimulated with peptidoglycan (PG) and lipopolysaccharide (LPS) were investigated. Methods : RAW 264.7 co-stimulated with LPS and PG were incubated with BA at concentrations of 25 and 50 µM. Incubation time was 18 h, 20 h, 22 h, 24 h, and 26 h. After incubation, the production of hydrogen peroxide in RAW 264.7 was measured with dihydrorhodamine 123 assay. Additionally, RAW 264.7 stimulated with PG were incubated with BA at concentrations of 25 and 50 µM for 24 h. After incubation, NO production was evaluated by griess reagent assay. Results : BA significantly inhibited hydrogen peroxide productions (p <0.05). In details, production of hydrogen peroxide in 'LPS and PG'-stimulated RAW 264.7 treated for 18 h with BA at concentrations of 25 and 50 µM was 91.27% and 89.22% of the control group treated with LPS and PG only, respectively; the production of hydrogen peroxide for 20 h was 92.19% and 90.58%, respectively; production of hydrogen peroxide for 22 h was 91.69% and 89.89%, respectively; production of hydrogen peroxide for 24 h was 92.4% and 90.19%, respectively; production of hydrogen peroxide for 26 h was 91.7% and 89.04%, respectively. Additionally, BA at the concentration of 50 and 100 µM significantly inhibited NO production in PG-induced RAW 264.7 (p <0.05). Conclusions : BA might have anti-oxidative activity related to its inhibition of hydrogen peroxide production in 'LPS and PG'-stimulated RAW 264.7 macrophages.

Quality comparison of non-thermal sterilized raw apple vinegar and commercial apple vinegar products

  • Sun Hwa Kim;Ji-Hyung Seo;Yong-Jin Jeong
    • Food Science and Preservation
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    • v.31 no.2
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    • pp.235-244
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    • 2024
  • A nonthermally sterilized raw apple vinegar was manufactured using an ultra-fine filtration process (0.2 ㎛ membrane filter) and its quality was comparable to commercially available vinegar products. First, using apple concentrate as a raw material, it was possible to produce non-thermal sterilized Using a two-stage fermentation process of alcohol and acetic acid fermentations, a non-thermally sterilized raw apple vinegar with pH 2.94 and an acidity of 6.20% was produced from an apple concentrate. The fermentation process increased the browning index significantly. However, the fundamental quality parameters of the non-thermal sterilized raw apple vinegar (A) with sterilized apple vinegar (B) did not differ significantly. The pH (2.92-2.95) of apple vinegar (A and B) was higher than that (pH 2.65-2.70) of commercial vinegar (C and D), and the total acidity, which is in the range of 6.20-6.21% and 6.53-6.90%, respectively, was higher in samples C and D than in samples A and B. However, four kinds of organic acids were detected in non-thermal sterilized raw apple vinegar (A), and its total organic acid content (6,245.00 mg%) was significantly higher than that of other samples (B, C, D) (p<0.05). In particular, malic acid content, as a main organic acid in apples, was very high in sample (A) (244.83 mg%) and sample (B) (210.21 mg%), compared to commercial products C (125.78 mg%) and D (86.90 mg%). The total polyphenol content and antioxidant activity of fermented apple vinegar (A, B) were more than twice as high as those of commercial products (C, D). Vinegar A had higher total polyphenol content than vinegar B. The above results suggest it is possible to manufacture and commercialize non-thermal sterilized raw apple vinegar with higher organic acid content and antioxidant properties using ultra-fine filtration.

Effect of Conjugated Linoleic Acid on Nuclear Factor-${\kappa}B$ Activation and Tumor Necrosis Factor-${\alpha}$ Production in RAW 264.7 Cells Exposed to High Concentration of Glucose (고농도의 당에 노출된 RAW 264.7 세포에서 conjugated linoleic acid의 TNF-${\alpha}$ 생산과 NF-${\kappa}B$의 활성 효과)

  • Lee, Minji;Kang, Byeong-Teck;Kang, Ji-Houn;Yang, Mhan-Pyo
    • Journal of Veterinary Clinics
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    • v.29 no.5
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    • pp.361-367
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    • 2012
  • Diabetes-related complications in human and veterinary medicine have been shown to be associated with hyperglycemia-induced inflammation. It has been recently suggested that the onset of insulin resistance may be caused by over-production of inflammatory cytokines such as tumor necrosis factor (TNF)-${\alpha}$ from immune cells. Conjugated linoleic acid (CLA) regulates inflammatory response through modulation of TNF-${\alpha}$ expression. The objective of this study was to examine the effect of CLA on nuclear factor kappaB (NF-${\kappa}B$) p65 binding activity, inhibitory kappaB ($I{\kappa}B$)-${\alpha}$ expression, and TNF-${\alpha}$ production from high glucose-treated RAW 264.7 cells. CLA was added to RAW cells that had been previously cultured with low or high concentration of glucose. The levels of TNF-${\alpha}$ protein in the culture supernatant of RAW cells exposed to high concentrations of glucose were higher than those of cells exposed to low concentrations of glucose. The treatment with the high concentration of glucose in RAW cells increased levels of NF-${\kappa}B$ p65 binding activity and the decreased $I{\kappa}B-{\alpha}$ expression when compared with those of low glucose. The treatments in combination with CLA and glucose (low and high) glucose in RAW cells increased TNF-${\alpha}$ production when compared with that glucose alone. These treatments with CLA increased TNF-${\alpha}$ production in high glucose-treated RAW cells than those with low glucose. These treatments of CLA also showed higher NF-${\kappa}B$ p65 binding activity and lower $I{\kappa}B-{\alpha}$ expression in high glucose than those in low glucose condition. This suggests that CLA can increase NF-${\kappa}B$ p65 binding activity and TNF-${\alpha}$ production from high glucose-treated RAW 264.7 cells and is likely to promote hyperglycemia-induced inflammation.

Trans-10, cis-12 Conjugated Linoleic Acid Modulates Tumor Necrosis Factor-${\alpha}$ Production and Nuclear Factor-${\kappa}B$ Activation in RAW 264.7 Macrophages Through Formation of Reactive Oxygen Species (RAW 264.7 세포에 있어 t10c12-CLA의 ROS를 통한 TNF-${\alpha}$ 생산 및 NF-${\kappa}B$ 활성 조절)

  • Park, So-Young;Kang, Byeong-Teck;Kang, Ji-Houn;Yang, Mhan-Pyo
    • 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.

The development of reclaimer automatic system for raw material (원료 Reclaimer 자동화 시스템 개발)

  • 박형근;문성룡
    • 제어로봇시스템학회:학술대회논문집
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    • 1996.10b
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    • pp.1276-1279
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    • 1996
  • Reclaimer in the raw material yard is being used to dig iron and coal so that they transfer to main blast furnace. A newly automatic system was developed and tested in the raw yard of Kwangyang iron making. The concept of the proposed system is based on the 3-dimensional detection of pile and auto-landing on the surface it.

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