• Title/Summary/Keyword: RNA degradation

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Isolation and Characterization of Aeromons hydrophila PBl6 and Properties of Synthetic Wastewater Degradation (Protease 생성균 Aeromonas hydrophila PB16의 분리 및 합성폐수처리능)

  • 박형수;양선영;김무훈;이종광;유용호;박두현
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.235-240
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    • 2002
  • Protease producing bacterium, PB16 was isolated from food processing wastewater sludge and paddy field soil samples and selected by the clear zone and enzyme activity test. The isolate was gram negative, rod type and its protease productivity was 6.49 U/ml. As a result of API20NE kit test and 16S rDNA sequencying, the isolated PB16 was identified as Aeromonas hydrophila (99%). The growth rate ($h^{-1}$) was 0.21 in synthetic waste water only and 0.26 in synthetic waste water containing vitamin and mineral using a bioscreen C. Synthetic wastewater removal rate was 59 and 87%, respectively after 1 and 3 day reaction (intial CODcr was 2,472 mg/l).

Tamoxifen Suppresses Clusterin Level through Akt Inactivation and Proteasome Degradation in Human Prostate Cancer Cells

  • Shim, Jae-Ho;Choi, Chang-Su;Lee, Eun-Chang;Kim, Mie-Young;Chun, Young-Jin
    • Biomolecules & Therapeutics
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    • v.17 no.1
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    • pp.25-31
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    • 2009
  • Clusterin is a heterodimeric sulfated glycoprotein and plays a role in many different types of cancer as a cell survival factor and helps cancerous cells to evade stress-induced apoptosis. To investigate whether the regulation of clusterin expression is involved in the mechanism of anticancer agent, we studied the effect of tamoxifen on clusterin expression in human prostate cancer PC-3 cells. Treatment of PC-3 cells with tamoxifen reduced cellular proliferation. Western blot analyses showed that treatment with tamoxifen suppressed clusterin expression in a concentration-dependent manner. Transfection with clusterin siRNA plasmid showed that clusterin is required for PC-3 cell survival. We found that tamoxifen resulted in a rapid decrease in the phosphorylation of Akt on Ser473 leading to prevent kinase activity. Expression of myristoylated Akt prevented tamoxifen-mediated clusterin downregulation. Interestingly, MG132, a wellknown proteasome inhibitor also recovered clusterin expression suppressed by tamoxifen. These data indicate that clusterin expression may be regulated by activation of Akt and ubiquitin-proteasome pathway plays an important role in tamoxifen-mediated clusterin suppression.

Inactivation of SMAD$_4$ Tumor Suppressor gene during Gastric Cancer Progression

  • Shin, Young-Kee
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2006.11a
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    • pp.19-24
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    • 2006
  • Mothers against decapentaplegic homolog 4 (SMAD4) is a tumor suppressor gene associated with gastrointestinal carcinogenesis. The aim of the present study was to characterize more precisely its role in the development and progression of human gastric carcinoma. In this study, using tissue microarray analysis of 283 gastric cancers and related lesions, we found loss of SMAD4 protein expression in the cytoplasm (36/114, 32%) and in the nucleus (46/114, 40%) of gastric cancer cells. The loss of nuclear SMAD4 expression in primary tumors correlated significantly with poor survival, and was an independent prognostic marker in multivariate analysis. We also found a substantial decrease in SMAD4 expression at both the RNA and protein level in several human gastric carcinoma cell lines. To identify the genetic and/or epigenetic mechanisms of altered SMAD4 expression in gastric carcinoma, loss of heterozygosity (LOH), promoter hypermethylation, and exon mutations were examined. We found that LOH (20/70, 29%) and promoter hypermethylation (4/73, 5%) were associated with the loss of SMAD4 expression. SMAD4 protein levels wore also affected in certain gastric carcinoma cell lines following incubation with Mc132, a proteasome inhibitor. Taken together, our results indicate that the loss of SMAD4, especially loss of nuclear SMAD4 expression, is involved in gastric cancer progression. The loss of SMAD4 in gastric carcinomas is due to several mechanisms, including LOH, hypermethylation, and proteasome degradation.

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Association between oropharyngeal microbiome and weight gain in piglets during pre and post weaning life

  • Bugenyi, Andrew Wange;Cho, Ho-Seong;Heo, Jaeyoung
    • Journal of Animal Science and Technology
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    • v.62 no.2
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    • pp.247-262
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    • 2020
  • Birth weight and subsequent weight gain is of critical importance in the survival and performance of piglets on a commercial swine farm setting. Oropharyngeal microbiome could influence immunity, and feeding behavior thus impacting health and weight gain. We used 16S rRNA gene sequencing to profile the composition and predicted metabolic functionality of the oropharyngeal microbiota in 8 piglets (4 with a birthweight ≤ 1.0 kg and 4 with a birthweight ≥ 1.7 kg) at 11, 26, and 63 days of age. We found 9 genera that were significantly associated with average daily gain (ADG) at 11 days (false discovery rate, FDR < 0.05) and 26 days of age (FDR < 0.1), respectively. The microbial functional profile revealed several pathways associated with ADG (FDR < 0.05). Among these, pathways related to degradation of catechols showed a positive association with ADG at 11, 26, and 63 days of age, implying a potential to breakdown the host-derived catecholamines. We also noted that pathways related to the biodegradation of nucleosides and nucleotides increased with ADG during the pre-weaning phase, while those involved in their biosynthesis decreased. Our findings provide insights into the oropharyngeal microbial memberships and metabolic pathways that are involved in a piglet's weight gain. Thus, providing a basis for the development of strategies aimed at improving weight gain in pigs.

Characterization of Diesel Degrading Enterobacter cancerogenus DA1 from Contaminated Soil

  • Kim, Sang-Jun;Joo, Gil-Jae
    • Korean Journal of Environmental Biology
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    • v.36 no.2
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    • pp.190-198
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    • 2018
  • The petroleum industry is an important part of the world economy. However, the massive exposure of petroleum in nature is a major cause of environmental pollution. Therefore, the microbial mediated biodegradation of petroleum residues is an emerging scientific approach used to resolve these problem. Through the screening of diesel contaminated soil we isolated a rapid phenanthrene and a diesel degrading bacterium identified as Enterobacter cancerogenus DA1 strain through 16S rRNA gene sequence analysis. The strain was registered in NCBI with an accession number MG270576. The optimal growth condition of the DA1 strain was determined at pH 8 and $35^{\circ}C$, and the highest degradation rate of the diesel was achieved at this condition. At the optimal condition, growth of the strain on the medium containing 0.05% phenanthrene and 0.1% of diesel-fuel was highest at 45 h and 60 h respectively after the incubation period. Biofilm formation was found significantly higher at $35^{\circ}C$ as compared to $30^{\circ}C$ and $40^{\circ}C$. Likewise, the lipase activity was found significantly higher at 48 h after the incubation compared to 24 h and 72 h. These results suggest that the Enterobacter cancerogenus DA1 could be an efficient candidate, for application through ecofriendly scientific approach, for the biodegradation of petroleum products like diesel.

Chitinase-producing Salinivibrio bacteria isolated from salt-fermented shrimp with antimicrobial and safety assessments

  • Le, Bao;Chung, Gyuhwa;Yang, Seung Hwan
    • Journal of Applied Biological Chemistry
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    • v.61 no.3
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    • pp.233-238
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    • 2018
  • Chitinases are glycosyl hydrolases which cleave the ${\beta}$-1,4 linkage of chitin into oligo or monomers of N-acetylglucosamine. These bacterial enzymes have been used for a wide range of applications in the food and pharmaceutical industries. In this study, we isolated two potential chitinolytic strains, BAO-01 and BAO-02, from salt-fermented shrimp, which were shown to belong to the genus Salinivibrio through genetic characterization using 16S rRNA. These isolates were gram-positive, rod-shaped, and non-spore forming. BAO-01 showed greater growth and chitinase activity than BAO-02 after the incubation at $37^{\circ}C$ for 4 days. Both strains grew on a wide range of carbon and nitrogen sources, pH values, temperatures, and salt levels. However, they showed minor biochemical differences. In addition, their antimicrobial activities against foodborne pathogens and antibiotic susceptibilities were evaluated. These Salinivibrio spp. did not show bioamine production, hemolytic activity, and mucin degradation. Therefore, the in vitro screening results suggested that these bacteria could be widely used as new candidates for chitin hydrolyzation and seafood fermentation.

Vitamin C Blocks TNF-${\alpha}$-induced NF-kB Activation and ICAM-1 Expression in Human Neuroblastoma Cells

  • Son, Eun-Wha;Mo, Sung-Ji;Rhee, Dong-Kwon;Pyo, Suhk-Neung
    • Archives of Pharmacal Research
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    • v.27 no.10
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    • pp.1073-1079
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    • 2004
  • Interactions of the cell adhesion molecules are known to play important roles in mediating inflammation. The proinflammatory cytokine, tumor necrosis factor-${\alpha}$(TNF-${\alpha}$), activates the NF-kB signaling pathway, which induces the expression of various genes, such as intercellular adhesion molecule-1 (ICAM-1). In this study, the effect of vitamin C on the ICAM-1 expression induced by TNF-${\alpha}$ in a human neuroblastoma cell line, SK-N-SH was investigated. Treatment with vitamin C resulted in the downregulation of the TNF-${\alpha}$-induced surface expression and ICAM-1 mRNA levels in a concentration-dependent manner. Moreover, a gel shift analysis indicated that vitamin C dose-dependently inhibited the NF-kB activation and IkB${\alpha}$ degradation induced by TNF-${\alpha}$. Taken together, these results suggest that vitamin C downregulates TNF-${\alpha}$- induced ICAM-1 expression via the inhibition of NF-kB activation.

Anti-Inflammatory Effect of the Extracts from Abeliophyllum distichum Nakai in LPS-Stimulated RAW264.7 Cells

  • Park, Gwang Hun;Park, Jae Ho;Eo, Hyun Ji;Song, Hun Min;Lee, Man Hyo;Lee, Jeong Rak;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.27 no.3
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    • pp.209-214
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    • 2014
  • In this study, we investigated whether A. distichum decreases the production of inflammatory mediators through downregulation of the NF-${\kappa}B$ and ERK pathway. Our data indicated that A. distichum leaf inhibits the overexpression of iNOS in protein and mRNA levels, and subsequently blocked LPS-mediated NO overproduction in RAW264.7 cells. A. distichum leaf inhibited $I{\kappa}B-{\alpha}$ degradation and p65 nuclear translocation, and subsequently suppressed transcriptional activity of NF-${\kappa}B$ in LPS-stimulated RAW264.7 cells. In addition, A. distichum leaf suppressed LPS-induced ERK1/2 activation by decreasing phosphorylation of ERK1/2. These findings suggest that A. distichum leaf shows anti-inflammatory activities through suppressing ERK-mediated NF-${\kappa}B$ activation in mouse macrophage.

NRROS Negatively Regulates Osteoclast Differentiation by Inhibiting RANKL-Mediated NF-κB and Reactive Oxygen Species Pathways

  • Kim, Jung Ha;Kim, Kabsun;Kim, Inyoung;Seong, Semun;Kim, Nacksung
    • Molecules and Cells
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    • v.38 no.10
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    • pp.904-910
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    • 2015
  • Negative regulator of reactive oxygen species (NRROS) is known to repress ROS generation in phagocytes. In this study, we examined the roles of NRROS in both osteoclasts and osteoblasts. Our results demonstrate that NRROS negatively regulates the differentiation of osteoclasts, but not osteoblasts. Further, overexpression of NRROS in osteoclast precursor cells attenuates RANKL-induced osteoclast differentiation. Conversely, osteoclast differentiation is enhanced upon siRNA-mediated knock-down of NRROS. Additionally, NRROS attenuates RANKL-induced $NF-{\kappa}B$ activation, as well as degradation of the NOX1 and NOX2 proteins, which are required for ROS generation. Based on our observations, we present NRROS as a novel negative regulator of RANKL-induced osteoclastogenesis.

A splice variant of human Bmal1 acts as a negative regulator of the molecular circadian clock

  • Lee, Jiwon;Park, Eonyoung;Kim, Ga Hye;Kwon, Ilmin;Kim, Kyungjin
    • Experimental and Molecular Medicine
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    • v.50 no.12
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    • pp.6.1-6.10
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    • 2018
  • Bmal1 is one of the key molecules that controls the mammalian molecular clock. In humans, two isoforms of Bmal1 are generated by alternative RNA splicing. Unlike the extensively studied hBmal1b, the canonical form of Bmal1 in most species, the expression and/or function of another human-specific isoform, hBmal1a, are poorly understood. Due to the lack of the N-terminal nuclear localization signal (NLS), hBMAL1a does not enter the nucleus as hBMAL1b does. However, despite the lack of the NLS, hBMAL1a still dimerizes with either hCLOCK or hBMAL1b and thereby promotes cytoplasmic retention or protein degradation, respectively. Consequently, hBMAL1a interferes with hCLOCK:hBMAL1b-induced transcriptional activation and the circadian oscillation of Period2. Moreover, when the expression of endogenous hBmal1a is aborted by CRISPR/Cas9-mediated knockout, the rhythmic expression of hPer2 and hBmal1b is restored in cultured HeLa cells. Together, these results suggest a role for hBMAL1a as a negative regulator of the mammalian molecular clock.