• Title/Summary/Keyword: P19 cells

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Screening for Biological Activity of Crude Extract and Bioactive Fractions from Brachyglottis monroi

  • Baek Seung Hwa;Lim Jin A;Kwag Jung Sook;Lee Hyun Ok;Chun Hyun Ja;Lee Jeong Ho;Perry Nigel B.
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.826-828
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    • 2003
  • The effects of crude extract and bioactive fractions from Brachyglottis monroi on biological activity were investigated. The crude ethanol extract inhibited the growth of the Gram positive bacterium Bacillus subtilis (ATCC Strain number 19659, 1 mm zone at 150 μg/disk) and the dermatophyte Trichophyton mentagrophytes (ATCC 28185, 2 mm zone at 150 μg/disk), and toxic to P388 tumor cells (IC50 23.96 μg/ml at 75 μg/disk). Cytotoxic activity was strongly showed by Fr. 6 (P388 IC50 19.67 μg/ml at 75 μg/disk).

Effect of random Shine-Dalarno sequence on the expression of Bovine Growth Hormone Gene in Escherichia coli (대장균에서 무작위 샤인-달가노 서열이 소성장호르몬 유전자 발현에 미치는 영향)

  • 나경수;나경수;백형석;이용세
    • Journal of Life Science
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    • v.10 no.4
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    • pp.422-430
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    • 2000
  • In order to search for the effects of Shine-Dalgarno (SD) sequence and nucleotide sequence of spacer region (SD-ATG) on bGH expression, oligonucleotides containing random SD sequences and a spacer region were chemically synthesized. The distance between SD region and initiation codon (ATG) was fixed to 9 nucleotides in length. The expression vectors have been constructed using pT7-1 vector containing a T7 promoter. Positive clones were screened with colony hybridization and named pT7A or pT7B plasmid series. The selected clones were confirmed by DNA sequencing and finally, 19 clones having various SD combinations were obtained. When bovine growth hormone was induced by IPTG in E. coli BL21(DE3), all cells harboring these plasmids produced a detectable level of bGH in western blot analysis. However, various SD sequences did not affect on bGH expression, indicating that the sequences of SD and the spacer region did not sufficiently destabilize mRNA secondary structure of bGH gene. Therefore, these results indicate that the disruption of mRNA secondary structure might be a major factor for regulating bGH expression in the translational initiation process.

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Degradation of Trichloroethylene by a Growth-Arrested Pseudomonas putida

  • Hahm, Dae-Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.1
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    • pp.11-14
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    • 1998
  • A toluene-oxidizing strain of Pseudomanas mendocina KR1 containing toluene-4-mono-oxygenase (TMO) completely degrades TCE with the addition of toluene as a co-substrate in aerobic condition. In order to construct in situ bioremediation system for TCE degradation without any growth-stimulating nutrients or toxic inducer such as toluene, we used the carbon-starvation promoter of Pseudomonas putida MK1 (Kim, Y. et al., J. bacteriol., 1995). Upon entry into the stationary phase due to the deprivation of nutrients, this promoter is strongly induced without further cell growth. The TMO gene cluster (4.5 kb) was spliced downstream of the carbon starvation promoter of Pseudomonas putida MK1, already cloned in pUC19. TMO under the carbon starvation promoter was not expressed in E. coli cells either in stationary phase or exponential phase. For TMO expression in Pseudomonas strains, tmo and carbon starvation promoter region were recloned into a modified broad-host range vector pMMB67HES which was made from pMMB67HE(8.9 kb) by deletion of tac promoter and lacIq (about 1.5 kb). Indigo was produced by TMO under the carbon starvation promoter in a Pseudomonas strain of post-exponential phase on M9 (0.2% glucose and 1mM indole) or LB. 18% of TCE was degraded in 14 hours after entering the stationary phase at the initial concentration of 6.6 ${\mu}$M in liquid phase.

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The Involvement of p38 MAPK and JNK Activation in Palmitic Acid-Induced Apoptosis in Rat Hepatocytes (Palmitic acid에 의한 간세포 사멸효과에 대한 p38 MAPK 및 JNK 관련성)

  • Bae, Chun-Sik;Park, Soo-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1119-1124
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    • 2009
  • Hyperlipidemia has been reported to be associated with the development of fatty liver. Palmitic acid, a major saturated fatty acid, is involved in the development of diverse diseases. The activation of mitogen activated protein kinases (MAPKs), such as Jun N-terminal kinase (INKs) and p38 MAPK is implicated in the apoptosis in diverse cells. Thus, this study was conducted to investigate the effects of palmitic acid on apoptosis and its relationship between JNK and p38 MAPK in cultured rat hepatocytes. In the present study, palmitic acid (>50 uM) decreased cell proliferation and increased lactate dehydrogenase activity in hepatocytes, which was blocked by the treatment of SP600125 (a JNK inhibitor) and SB203580 (a p38 MAPK inhibitor). Indeed, palmitic acid decreased Bcl-2 expression but increased Bax expression in rat hepatocytes, which was blocked by the treatment of SP600125 and SB203580. In addition, palmitic acid decreased glutathione (GSH) content and increased lipid peroxide formation, which was blocked by the treatment of SP600125 and SB203580. Western immunoblotting analysis also revealed that palmitic acid increased JNK and p38 MAPK. In conclusion, palmitic acid induced apoptosis through oxidative stress via JNK and p38 MAPK activation in rat hepatocytes.

Advantages of Restoring miR-205-3p Expression for Better Prognosis of Gastric Cancer via Prevention of Epithelial-mesenchymal Transition

  • Zhang, Zhen;He, Xujun;Xu, Ji;Zhang, Genhua;Yang, Yue;Ma, Jie;Sun, Yuanshui;Ni, Haibin;Wang, Fengyong
    • Journal of Gastric Cancer
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    • v.20 no.2
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    • pp.212-224
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    • 2020
  • Purpose: miR-205 is a tumor suppressor and plays an important role in tumor invasiveness. However, the role of miR-205 in human gastric cancer (GC) epithelial-mesenchymal transition (EMT) remains unclear. The aim of this study was to investigate the molecular mechanism of miR-205 in the regulation of EMT in GC invasion. Materials and Methods: Quantitative polymerase chain reaction (qPCR) was used to detect the expression of miR-205 in GC. Further, the correlation between the pathological parameters and prognosis of GC was statistically analyzed. A transwell model was used to evaluate the effect of miR-205-3p on the invasion and migration of GC cells. qPCR, western blotting, and luciferase assay were performed to analyze the relationship and target effects between miR-205-3p and the expression of zinc finger electron box binding homologous box 1 (ZEB1) and 2 (ZEB2). Results: We found that the levels of miR-205-3p were significantly lower (P<0.05) in GC tissues than in matched normal tissues. Additionally, the expression of miR-205-3p was related to the tumor invasion depth, lymph node metastasis, lymph node invasion, and tumor, node, metastasis stage. Patients with lower miR-205-3p expression levels in the tumors had a poorer prognosis. The in vitro assays indicated that miR-205-3p could affect the invasion ability and EMT of GC cells by targeting the expression of both ZEB1 and ZEB2. Conclusions: miR-205-3p promotes GC progression and affects the prognosis of patients by targeting both ZEB1 and ZEB2 to directly influence EMT.

Effect of Addition of Enzyme-Resistant Rice RS3 on Quality and Textural Characteristics of Madeleine (효소저항성 쌀전분의 첨가가 마들렌의 품질 및 텍스처 특성에 미치는 영향)

  • Kim, Wan-Soo
    • Korean Journal of Human Ecology
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    • v.19 no.1
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    • pp.191-201
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    • 2010
  • This study attempted to examine the application of retrograded starch (RS3) isolated from rice flour into Madeleine which is easy to make, supply enough energy and micro nutrients with adequate drinks, and prevent an adult disease. This could be a popular food to anyone regardless of age and gender who avoid rice and become high value-added, processed rice foods. For this, control Madeleine was made from wheat flour and an experimental one was made from 5 or 10% rice RS3 addition as well as wheat flour. Four different types of rice were produced from Premium Ho-Pyong Rice, that is, dry milled rice flour(RFD), soaked for 8 hours and milled, followed by air-dried rice flour(RFW), rice starch(RST), and retrograded rice starch or enzyme-resistant starch(RS3). The results found were as follows: Proximate compositions were decreased with soaking to make RFW, RST and RS3, compared to RFD. RS3 had the highest L, +a and ${\Delta}E$ with the lowest +b, changing it to a dark color, explaining the need for heat control during processing. At $80^{\circ}C$, the swelling power was shown in the order of RST>RFW>RFD>RS3 and the solubility of RS3 was the highest. There were significant differences in viscosities of peak, trough, cold, breakdown and total setback of all rice samples using RVA (p<0.001). Due to the pH of RS3, the Madeleine batter became acidic (p<.01) and expanded, resulting in more air cells and open texture. With an increasing RS3 level in Madeleine, several textural attributes among 'fresh' and 'stored at room temperature' Madeleine samples were significantly different by using Texture Analyzer. While the addition of RS3 in Madeleine did not significantly affect the sensory evaluation, indicating RS3 isolated from rice as a beneficial ingredient for processed rice products.

Yam (Dioscorea batatas) Root and Bark Extracts Stimulate Osteoblast Mineralization by Increasing Ca and P Accumulation and Alkaline Phosphatase Activity

  • Kim, Suji;Shin, Mee-Young;Son, Kun-Ho;Sohn, Ho-Yong;Lim, Jae-Hwan;Lee, Jong-Hwa;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.19 no.3
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    • pp.194-203
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    • 2014
  • Yam (Dioscorea batatas) is widely consumed as functional food for health promotion mainly in East Asia countries. We assessed whether yam root (tuber) or bark (peel) extracts stimulated the activity of osteoblasts for osteogenesis. MC3T3-E1 cells (mouse osteoblasts) were treated with yam root extracts (water or methanol) (study I) or bark extracts (water or hexane) (study II) within $0{\sim}10{\mu}g/mL$ during the periods of osteoblast proliferation (5~10 day), matrix maturation (11~15 day) and mineralization (16~20 day) as appropriate. In study I, both yam root water and methanol extracts increased cell proliferation as concentration-dependent manner. Cellular collagen synthesis and alkaline phosphatase (ALP) activity, both the indicators of bone matrix protein and inorganic phosphate production for calcification respectively, were also increased by yam root water and methanol extract. Osteoblast calcification as cell matrix Ca and P accumulation was also increased by the addition of yam root extracts. In study II, yam bark extracts (water and hexane) increased osteoblast proliferation and differentiation, as collagen synthesis and ALP activity and osteoblast matrix Ca and P deposition. The study results suggested that both yam root and bark extracts stimulate osteogenic function in osteoblasts by stimulating bone matrix maturation by increasing collagen synthesis, ALP activity, and matrix mineralization.

Evaluation of the Synergistic Effect of Mixed Cultures of White-Rot Fungus Pleurotus ostreatus and Biosurfactant-Producing Bacteria on DDT Biodegradation

  • Purnomo, Adi Setyo;Ashari, Khoirul;Hermansyah, Farizha Triyogi
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1306-1315
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    • 2017
  • DDT (1,1,1-trichloro-2,2-bis(4-chlorophenyl) ethane) is one of the organic synthetic pesticides that has many negative effects for human health and the environment. The purpose of this study was to investigate the synergistic effect of mixed cutures of white-rot fungus, Pleurotus ostreatus, and biosurfactant-producing bacteria, Pseudomonas aeruginosa and Bacillus subtilis, on DDT biodegradation. Bacteria were added into the P. ostreatus culture (mycelial wet weight on average by 8.53 g) in concentrations of 1, 3, 5, and 10 ml ($1ml{\approx}1.25{\times}10^9$ bacteria cells/ml culture). DDT was degraded to approximately 19% by P. ostreatus during the 7-day incubation period. The principal result of this study was that the addition of 3 ml of P. aeruginosa into P. ostreatus culture gave the highest DDT degradation rate (approximately 86%) during the 7-day incubation period. This mixed culture combination of the fungus and bacteria also gave the best ratio of optimization of 1.91. DDD (1,1-dichloro-2,2-bis(4-chlorophenyl) ethane), DDE (1,1-dichloro-2,2-bis(4-chlorophenyl) ethylene), and DDMU (1-chloro-2,2-bis(4-chlorophenyl) ethylene) were detected as metabolic products from the DDT degradation by P. ostreatus and P. aeruginosa. The results of this study indicate that P. aeruginosa has a synergistic relationship with P. ostreatus and can be used to optimize the degradation of DDT by P. ostreatus.

Human $CD103^+$ dendritic cells promote the differentiation of Porphyromonas gingivalis heat shock protein peptide-specific regulatory T cells

  • Kim, Myung-Jin;Jeong, Eui-Kyong;Kwon, Eun-Young;Joo, Ji-Young;Lee, Ju-Youn;Choi, Jeomil
    • Journal of Periodontal and Implant Science
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    • v.44 no.5
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    • pp.235-241
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    • 2014
  • Purpose: Regulatory T cells (Tregs), expressing CD4 and CD25 as well as Foxp3, are known to play a pivotal role in immunoregulatory function in autoimmune diseases, cancers, and graft rejection. Dendritic cells (DCs) are considered the major antigen-presenting cells (APCs) for initiating these T-cell immune responses, of which $CD103^+$ DCs are derived from precursor human peripheral blood mononuclear cells (PBMCs). The aim of the present study was to evaluate the capacity of these PBMC-derived $CD103^+$ DCs to promote the differentiation of antigen-specific Tregs. Methods: Monocyte-derived DCs were induced from $CD14^+$ monocytes from the PBMCs of 10 healthy subjects. Once the $CD103^+$ DCs were purified, the cell population was enriched by adding retinoic acid (RA). Peptide numbers 14 and 19 of Porphyromonas gingivalis heat shock protein 60 (HSP60) were synthesized to pulse $CD103^+$ DCs as a tool for presenting the peptide antigens to stimulate $CD3^+$ T cells that were isolated from human PBMC. Exogenous interleukin 2 was added as a coculture supplement. The antigen-specific T-cell lines established were phenotypically identified for their expression of CD4, CD25, or Foxp3. Results: When PBMCs were used as APCs, they demonstrated only a marginal capacity to stimulate peptide-specific Tregs, whereas $CD103^+$ DCs showed a potent antigen presenting capability to promote the peptide-specific Tregs, especially for peptide 14. RA enhanced the conversion of $CD103^+$ DCs, which paralleled the antigen-specific Treg-stimulating effect, though the differences failed to reach statistical significance. Conclusions: We demonstrated that $CD103^+$ DCs can promote antigen-specific Tregs from naive T cells, when used as APCs for an epitope peptide from P. gingivalis HSP60. RA was an effective reagent that induces mature DCs with the typical phenotypic expression of CD103 that demonstrated the functional capability to promote antigen-specific Tregs.

The Effects of Bujeong hangamtang on antitumor Immune Response (부정항암탕(扶正抗癌湯)이 항종양(抗腫瘍) 면역반응(免疫反應)에 미치는 영향(影響))

  • Im, Mi-Yang;Moon, Sook-Jea;Moon, Gu;Won, Jin-Hee;Jeon, Byung-Hoon
    • The Journal of Korean Medicine
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    • v.19 no.1
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    • pp.234-250
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    • 1998
  • Bujeonghangamtang(扶正抗癌湯) has been used for cure of tumor as a traditional medicine without any experimental evidence to support the rational basis for its clinical use. This study was carroed out to evaluate the possible therapeutic or antitumoral effects of Bujeonghangamtang extract against tumor, and to carry out some mechanisms responsible for its effect. Some kinds of tumor were induced by .the typical application of 3-methylcholanthrene (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(Sl80 cells). Treatment of the Bujeonghangamtang water-extract (dailly 1mg/mouse, i. p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 20 days. Against squamous cell carcinoma induced by MCA, Bujeonghangamtang decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Bujeongmngamtang also significantly suppressed the development of 3LL cell and S180 cell-implanted tumors in occurence-frequency and their size, and some developed tumors were regressed by the continuous treatment of Bujeonghangamtang extract into TBM. In vitro, treatment of Bujeonghangamtang extract had no effect on the growth of some kinds of cell line such as FsaII, A431 strain but significantly inhibited the proliferation of 3LL, S180 cells and augmented the DNA synthesis of mitogen-activated lymphocytes. Bujeonghangamtang also stimulated the migrative ability of leukocyte, the MIF and IL-2 production of T lymphocytes, but not IL 6 production of B cells. Bujeonghangamtang-administration to mice enhanced NK cells attivities. These results demonstrated that Bujeonghangamtang extract exhibited a significant prophylactic benefits against tumors and its antitumor activity was manifested depending on the type of tumor cells. And these results also suggested that effect of Bujeonghangamtang might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

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