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A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

Plant regeneration and soil acclimatization through photoautotrophic culture from leaf explant of a rare species in Sedum tosaense Makino (희귀수종인 주걱비름(Sedum tosaense Makino)의 잎절편으로부터 기내 식물체 재분화 및 광독립배양을 통한 토양순화)

  • Ko, Myoung-Suk;Bae, Kee Hwa;Song, Gwanpil;So, In Sup
    • Journal of Plant Biotechnology
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    • v.40 no.2
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    • pp.79-87
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    • 2013
  • The aim of this study was to establish plant regeneration from leaf explants of Sedum tosaense Makino, which is globally rare and endangered species. The leaf explants of S. tosaense were cultured on the MS medium supplemented with different concentration of BA and NAA for callus induction. Callus induction was showed the highest (100%) on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA. The highest number of shoots were regenerated when callus were cultured on MS medium containing $2.0mg{\cdot}L^{-1}$ BA and $1.0mg{\cdot}L^{-1}$ NAA for 5 weeks. The axillary bud were cultured on the MS media supplemented with combination of BA and NAA for in vitro propagation. The highest number of adventitious shoot (7.9 per explants) formed at $1.0mg{\cdot}L^{-1}$ NAA and $2.0mg{\cdot}L^{-1}$ BA. For rooting, MS medium supplemented with or without $2.0g{\cdot}L^{-1}$ activated charcoal was tested. The optimal results were observed using MS medium supplemented with $2.0g{\cdot}L^{-1}$ activated charcoal, on which 85.7 (No. of root), 4.6 cm (length of root). 1,200 ppm $CO_2$ and 350 ppm $CO_2$ were supplied for make certain the effects of $CO_2$ on pre-acclimatization by photoautotrophic culture. 1,200 ppm $CO_2$ treatment was established higher than 350 ppm $CO_2$ treatment. Soil acclimatization of in vitro plantlets was the best in mixture soil consisted of peat moss and perlite with 100% survival rate and they showed the maximum growth.

Fructus Sophorae Enhances the Production of Prostaglandin E2 and Tumor Necrosis Factor-α through Activation of MAPKs and PI3K/AKT Signaling Pathways in Murine Macrophages (대식세포에서 MAPKs 및 PI3K/AKT 신호전달계 활성을 통한 괴각 추출물의 prostaglandin E2 및 tumor necrosis factor-α 생성의 촉진)

  • Kang, Young-Soon;Han, Min Ho;Lee, Moon Hee;Hong, Su Hyun;Park, Heungsik;Jung, Jae-Chul;Lee, Jeongrai;Lee, Eun-Woo;Kang, Kyung Hwa;Kim, Cheol Min;Kim, Byung-Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.23 no.11
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    • pp.1397-1403
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    • 2013
  • Fructus Sophorae, the dried ripe fruit of Styphnolobium japonicum (L.), is an herbal ingredient used in traditional Oriental medicine. This study was carried out to investigate the effects of Fructus Sophorae extracts (FSE) on immune modulation in a murine RAW 264.7 macrophage model. As immune response parameters, the production of prostaglandin $E_2$ ($PGE_2$) and tumor necrotic $factor-{\alpha}$ ($TNF-{\alpha}$) were evaluated. Our data revealed that FSE increased the macrophage activation and the production of $PGE_2$ and $TNF-{\alpha}$, which was consistently correlated with upregulation of cyclooxygenase-2 (COX-2) and $TNF-{\alpha}$ expression at both transcriptional and translational levels. On comparative cytokine protein array, FSE significantly increased several cytokines, which was associated with phosphorylation of mitogen- activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), p38 MAPK and c-Jun N-terminal kinase (JNK), and Akt in RAW 264.7 cells. However, each inhibitor of these molecules attenuated the FSE-induced $PGE_2$ production. These results indicate that FSE activated macrophages through the activation of MAPKs and phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathways in RAW 264.7 macrophages. These findings suggest that FSE may provide a promising source of an immunoenhancing agent.

Effect of the Flavonoid Luteolin for Dextran Sodium Sulfate-induced Colitis in NF-${\kappa}B^{EGFP}$ Transgenic Mice (Dextran Sodium Sulfate 유발 장염 모델에서 루테올린의 치료효과)

  • Jang, Byung-Ik
    • Journal of Yeungnam Medical Science
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    • v.23 no.1
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    • pp.26-35
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    • 2006
  • Background: Luteolin, a flavone found in various Chinese herbal medicines is known to possess anti-inflammatory properties through its ability to inhibit various proinflammatory signaling pathways including NF-${\kappa}B$ and p38 MAPK. In this study, we investigated the potential therapeutic effect of luteolin on dextran sodium sulfate (DSS)-induced colitis. Materials and Methods: We used a transgenic mouse model expressing the enhanced green fluorescent protein (EGFP) under the transcriptional control of NF-${\kappa}B$ $cis$-elements. C57BL/6 NF-${\kappa}B^{EGFP}$ mice received 2.5% DSS in their drinking water for six days in combination with daily luteolin administration (1mg/kg body weight, 0.1ml vol, intragastric) or vehicle. NF-${\kappa}B$ activity was assessed macroscopically with a Charge-Coupled Device (CCD) camera and microscopically by confocal analysis. Results: A significant increase in the Disease Activity Index (DAI), histological score (p<0.05), IL-12 p40 secretion in colonic stripe culture (p<0.05) and EGFP expression was observed in luteolin and/or DSS-treated mice compared to water-treated mice. Interestingly, a trend toward a worse colitis (DAI, IL-12p40) was observed in luteolin-treated mice compared to non-treated DSS-exposed mice. In addition, EGFP expression (NF-${\kappa}B$ activity) strongly increased in the luteolin-treated mice compared to control mice. Confocal microscopy showed that EGFP positive cells were primarily lamina propria immune cells. Conclusions: These results suggest that luteolin is not a therapeutic alternative for intestinal inflammatory disorders derived for primary defects in barrier function. Thus, therapeutic intervention targeting these signaling pathways should be viewed with caution.

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Effect of the Gibberellin Treatment on Enlargement and Mature Promotion in 'Niitaka' Pear (Pyrus pyryfolia L.) (지베렐린이 '신고'배의 비대와 성숙촉진에 미치는 영향)

  • Kim, Jong-Gook;Lee, Chang-Hoo
    • Journal of agriculture & life science
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    • v.43 no.2
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    • pp.23-30
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    • 2009
  • The object of this study was to evaluate the effect of the gibberellin treatment on fruit enlargement and ripening promotion in 'Niitaka' pear(Pyms pynfolia). Fruit weight was similar between $GA_{4+7}$ treatments and the gibberellin-paste control, but fairly increased of fruit weight compared to non-treatment Most effective time for $GA_{4+7}$ treatment to increase diameter and length was on 35 and 40 days after full bloom, respectively. Shape index was similar in all treatments. Fruit enlargement at the period of early growth, $GA_{4+7}$ 2.4% treatment was remarkably effective than $GA_{3}+GA_{4+7}$ 2.7% treatment or non-treatment However at the period of maturity, $GA_{4+7}$ 2.4% treatment and $GA_{3}+GA_{4+7}$ 2.7% treatment showed little differences in fruit enlargement and coloring. In maturing promotion effect, young fruit treated with $GA_{3}+GA_{4+7}$ showed similar fruit coloring to ethephon treatment on 35 days after full bloom, and both of those treatments promoted fruit coloring than non- treatment about 6 days. $GA_{3}+GA_{4+7}$ treatments resulted higher solid content and decreased acidity than non-treatment However, there was no differences in fruit hardness comparing to non-treatment As a result, gibberellin was most effective in fruit enlargement, so as all $GA_{3}+GA_{4+7}$ treatments were more effective on fruit enlargement than ethephon treatment or non-treatment Specifically, when $GA_{3}+GA_{4+7}$ was treated 35days after full bloom of flower, the diameter, the length and the shape of fruit index were best, and fruit coloring was good as well.

Effects of Storage Condition, Storage Period, and Priming on Seed Germination of Corylopsis coreana (저장방법 및 priming 처리가 히어리 종자 발아에 미치는 영향)

  • Kim, Hyoung Deug;Kim, Hong Lim;Kwack, Yong Bum;Choi, Young Hah;Lee, A Rong
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.4
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    • pp.266-270
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    • 2010
  • Corylopsis coreana is an endangered Korean native plants. This is one of the genus that have high ornamental value for flowering plants available for garden shrub, bonsai, and pot plants. In this study, the methods to encourage seed germination rate were investigated for its ornamental uses. The germination rate of Corylopsis coreana seeds stored under dry-cold condition was very low, 12%, 12%, 8%, and 10%after 40, 70, 85, and 100 days storage respectively. But the germination rate of Corylopsis coreana seeds stored under wet-cold condition was higher than these, 20%, 54%, 78%, and 96% after 40, 70, 85, and 100 days storage respectively. Dry seeds sowed directly without $GA_3$ treatment showed no germination regardless of storage type(cold or room temp.) or storage periods. On the other hand, the soaking treatment with $GA_3$ 50~500 ppm for 24 hours was very effective to increase the germination rate. The most effective $GA_3$ levels was different by storage type(cold or room temp.) and storage periods. But the effect of $GA_3$ was decreased by prolonging of the storage period. Soaking treatment with $Ca(NO_3)_2$ 5, 10, 20 mM, $KNO_3$ 5, 10, 20 mM for 24 hours showed no effect.

New Whitening Agent : Selina-4(14), 7(11)-dien-8-one (신규 미백제 : Selina-4(14), 7(11)-dien-8-one)

  • Kim, Cheong-Taek;Chang, Yun-Hee;Lee, Sang-Hwa;Kang, Sang-Jin;Cho, Wan-Goo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.1 s.49
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    • pp.17-23
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    • 2005
  • We had previously reported that Selina (selina-4(14), 7(11)-dien-8-one) was isolated from methanol extract of Afractylodes rhizome and has strong whitening activity in B16 melanoma cells. In this report, we demonstrated its action mechanism in melan-a cells, non-tumorigenic melanocytes. We also investigated the clinical efficacy of cosmetic preparation containing Selina. Selina reduced the melanin synthesis of Melan-a cells by $50\%$ at a concentration of $10 {\mu}g/mL$ without any apparent cytotoxicity. We also found that the treatment of cells with Selina decreased tyrosinase activity by $60\%$ at a concentration of $10 {\mu}g/mL$ but Selina was not a direct inhibitor of tyrosinase activities. To elucidate the action mechanism of Selina, we investigated the changes in mRNA and protein level of tyrosinase, TRP-1 and TRP-2 using RT-PCR and western blotting, respectively. As a result, the mRNA and protein level of tyrosinase were markedly reduced at $10 {\mu}g/mL$ of Selina without any effect on TRP-1 and TRP-2. These results suggest that Selina exerts its whitening effect mainly through regulating expression of tyrosinase. A 7 week-clinical trial using formulation containing $0.2\%$ selina-4(14), 7(11)-dien-8-one with 20 volunteers resulted in statistically significant whitening effect (p < 0.05), without any adverse effect. Based on these results, Selina (selina-4(14), 7(11)-dien-8-one) can be s useful and safe ingredient for the cleanness and brightness of skin.

Quality Characteristics and Inhibition Activity against Helicobacter pylori KCCM 40449 of Liquorice Yogurts Manufactured by Exopolysaccharide Producing Lactic Acid Bacteria (Exopolysaccharide 생성 유산균을 이용한 감초 추출물 첨가 Yoghurt의 품질특성 및 Helicobacter pylori KCCM 40449 억제활성)

  • Jung, Seung-Won;Kim, Cheol Woo;Lee, Su Han
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.346-354
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    • 2011
  • This study was carried out to fortify the antimicrobial activity of yoghurt by adding liquorice extract to it. The liquorice extracts (1 mg/mL) showed relatively high antibacterial activity against H. pylori KCCM 40449 (p < 0.05). The solvent liquorice extracts of minimal inhibitory concentrations (MIC) against H. pylori KCCM 40449 were 25- 100 ${\mu}g$/mL. Lactobacillus amylovorus DU-21 with high EPS production ability were inoulated to milk after the addition of different amounts of liquorice extracts (0.0%, 0.05%, 0.1% and 0.2%). The physico-chemical characteristics of yoghurts added with liquorice extracts were examined. The initial pH, titratable acidity, viscosity and viable cell counts of the yoghurt added liquorice extracts were 3.41-3.51, 1.021-1.091%, 1,686-1,930 cp and 9.41-9.38 Log CFU/mL, respectively. The viscosity and syneresis of yoghurt were better than that of the control. Antimicrobial activity against H. pylori KCCM 40449 increased with increasing addition of liquorice extract. However, the sensory score of yoghurt added with different amounts of liquorice extracts was lower than that of the control (p < 0.05). As a result of the sensory evaluations, the flavor, taste, texture, color and overall acceptability of the yoghurt with 0.05% liquorice extract were found to be much better than those of the other groups (p < 0.05). Overall, the optimal amount of liquorice extract added in the manufacture of yoghurt was 0.05% of the total weight. Further studies on increment of antimicrobial activity and palatability of liquorice extract added yoghurt are necessary.

Inhibitory Effects of Chrysanthemum boreale Makino on 3T3-L1 Preadipocyte Differentiation and Down-regulation of Adipogenesis and Lipogenesis (산국(Chrysanthemum boreale Makino) 꽃 유래 에센셜오일(Essential oil)이 지방세포 분화 및 지방생성에 미치는 영향)

  • Hwang, Dae Il;Choi, In-Ho;Kim, Do Yoon;Park, Soo Min;Kim, Ha Bin;Li, YaLi;Lee, Hwan Myung
    • Journal of Life Science
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    • v.29 no.3
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    • pp.332-336
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    • 2019
  • Obesity is associated with an increased risk of many diseases including type 2 diabetes mellitus, hypertension, and hyperlipidemia. The flowers of Chrysanthemum boreale have been used as traditional medicines for the treatment of diseases such as obesity and type 2 diabetes mellitus. This study aimed to evaluate the effect of C. boreale Makino flower essential oil (CFEO) on adipocyte differentiation using preadipocyte cell line 3T3-L1. CFEO at concentrations between 0.1 and $5{\mu}g/ml$ did not affect 3T3-L1 cell viability. A CFEO concentration of between 0.1 and $1{\mu}g/ml$ significantly inhibited lipid accumulation during MDI-induced differentiation in 3T3-L1 cells in a dose-dependent manner, reaching a maximal level at $1{\mu}g/ml$ ($28.94{\pm}2.01%$; approximately 30% of control treated with MDI alone). Western blot analysis revealed that CFEO concentrations between 0.1 and $1{\mu}g/ml$ suppressed the activations of three adipogenic transcription factors in the MDI-stimulated 3T3-L1 cells: peroxisome proliferator-activated receptor ${\gamma}$; CCATT/enhancer binding protein ${\alpha}$; and sterol regulatory element binding protein-1. Moreover, the expressions of lipogenic enzymes, acetyl-CoA carboxylase, and fatty acid synthase were also inhibited by treatment with CFEO between 0.1 and $1{\mu}g/ml$. CFEO may therefore be a promising functional material for obesity prevention.

Anti-inflammatory Effect of Heat-Killed Enterococcus faecalis, EF-2001 (열처리 사균체 엔테로코커스 패칼리스 EF-2001의 항염증 효과)

  • Choi, Moon-Suk;Chang, Sang-Jin;Chae, Yuri;Lee, Myung-Hun;Kim, Wan-Joong;Iwasa, Masahiro;Han, Kwon-Il;Kim, Wan-Jae;Kim, Tack-Joong
    • Journal of Life Science
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    • v.28 no.11
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    • pp.1361-1368
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    • 2018
  • Inflammation is the most common condition in the human body. Tissue damage triggers inflammation, together with vasodilation and increased blood flow at the inflamed site, resulting in edema. Inflammatory responses are also triggered by lipopolysaccharide (LPS), a Toll-like receptor Enterococcus faecalis, a gram-positive organism, has been reported to possess immunomodulatory and preventive activities; however, its use may present risks of sepsis and other systemic infections. Heat-killed Enterococcus faecalis (EF-2001) has been reported to induce antitumor activity, but its effects on inflammation are not known. In the present study, we investigated the effect of EF-2001 on LPS-induced macrophage inflammatory responses. EF-2001 treatment reduced nitric oxide (NO) production, indicating suppression of inflammatory reactions. EF-2001 showed no cytotoxicity in macrophages. Further investigation of the anti-inflammatory mechanism of EF-2001 indicated that EF-2001 reduced the LPS-induced expression of inducible nitric oxide synthase and cyclooxygenase-2. EF-2001 also reduced f the LPS induction of several inflammatory molecules involved in the nuclear factor-${\kappa}B$ ($NF-{\kappa}B$) and mitogen-activated protein kinase pathways, including ERK, JNK, and p38 phosphorylation, in a concentration-dependent manner. Additionally, EF-2001 inhibited Akt phosphorylation and increased the expression of the inhibitory ${\kappa}B$ ($I{\kappa}B$) protein, an inhibitor of $NF-{\kappa}B$. EF-2001 also inhibited the nuclear translocation of p65. These results suggest that EF-2001 has anti-inflammatory properties and may be useful for treating inflammatory diseases.