• Title/Summary/Keyword: 6S RNA

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Inhibitory effects of Sargassum horneri extract against endoplasmic reticulum stress in HepG2 cells (괭생이 모자반 추출물의 소포체 스트레스 억제 효능)

  • Park, Sora;Thomas, Shalom Sara;Cha, Youn-Soo;Kim, Kyung-Ah
    • Journal of Nutrition and Health
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    • v.53 no.6
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    • pp.583-595
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    • 2020
  • Purpose: This study examined the effects of Sargassum horneri extracts on palmitic acid (PA)-induced endoplasmic reticulum (ER) stress in HepG2 cells. Methods: HepG2 cells were treated with varying concentrations of S. horneri extract or PA, and the cell viability was measured by water soluble tetrazolium salts analysis. The effective induction of ER stress and the effects of S. horneri were investigated through an examination of the ER stress-related genes, such as activating transcription factor 4 (ATF4), X-box binding protein (XBP1s), C/EBP homologous protein (CHOP), and 78-kDa glucose-regulated protein (GRP78) by quantitative reverse transcription polymerase chain reaction. The expression and activation levels of unfolded protein response (UPR) associated proteins, such as inositol-requiring enzyme-1α (IRE1α), eukaryotic translation initiation factor 2 alpha submit (eIF2α), and CHOP were examined by western blot analysis. Results: The treatment with PA increased the expression of UPR associated genes significantly and induced ER stress in a 12-hour treatment. Subsequent treatment with S. horneri reduced mRNA expression of ATF4, GRP78, and XBP1s. In addition, the protein levels of phosphate (p)-IRE1α, p-elF2α, and CHOP were also reduced by a treatment with S. horneri. An analysis of sirtuin (SIRT) mRNA expression in the S. horneri and PA-treated HepG2 cells showed that S. horneri increased the levels of SIRT2, SIRT6, and SIRT7, which indicates a possible role in reducing the expression of ER stress-related genes. Conclusion: These data indicate that S. horneri can exert an inhibitory effect on ER stress caused by PA and highlight its potential as an agent for managing various ER stress-related diseases.

Inhibition of Porcine Endogenous Retrovirus Expression by RNA Interference (RNA 간섭을 통한 Porcine Endogenous Retrovirus의 발현 억제)

  • Lee, Hyun-A;Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.181-187
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    • 2006
  • In recent years the number of patients waiting for organ transplantation has greatly outpaced the supply of human organs available, which leads to a renewed interest in pig-to-human xenotransplantation as an alternative. However, one of the biggest barriers in the xenotransplantation is presence of porcine endogenous retroviruses(PERV) that can infect human cells. In this study, to present a possible solution for this problem we tried to inhibit expression of PERVs using shRNAs(short hairpin RNA) at the level of RNA synthesis and virus release. The shRNA targeting the sequence of PERV A, B type was cloned into pSIREN-RetroQ vector under the control of polymerase-III U6-RNA gene promoter. Quantitative real-time PCR was performed to detect my alterations in mRNA production of PERV A, B targeted by the shRNA in each done. Depending on the target sequence of the shRNA, the transcription of PERV was decreased to as much as 4% and the number of progeny viruses was reduced to less than 1/200,000. Transgenic pigs producing such shRNAs may result in a highly reduced PERV expression in cells and organs, which is a prerequisite for safe xenotransplantations.

DIFFERENTIAL EXPRESSION OF ORNITHINE DECARBOXYLASE AND HA-RAS CELLULAR ONCOGENE DURING DEVELOPMENT OF THE FEMALE RAT

  • Baik, M.G.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.1
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    • pp.119-124
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    • 1994
  • Experiments were performed to determine age-associated changes in ornithine decarboxylase (ODC) gene and Ha-ras cellular oncogene expression in tissues of female rats. In the kidney, ODC mRNA levels did not show age-associated changes, while ODC enzyme activities were decreased with advancing age from 3 to 10 months. These results suggest that post-transcriptional mechanism (s) are involved in the age-dependent decrease in renal ODC enzyme activity. In addition, we found no correlation between testosterone-induced renal ODC expression and DNA methylation pattern. Ha-ras mRNA levels in brain decreased as animals aged from 3 to 6 months, while renal Ha-ras mRNA levels were not influenced by age. Results demonstrate the age-dependent expression of Ha-ras in a tissue-specific manner.

Knockdown of cytosolic NADP+-dependent isocitrate dehydrogenase enhances MPP+-induced oxidative injury in PC12 cells

  • Yang, Eun-Sun;Park, Jeen-Woo
    • BMB Reports
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    • v.44 no.5
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    • pp.312-316
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    • 2011
  • 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and its toxic metabolite 1-methyl-4-phenylpyridium ion (MPP$^+$) have been shown to induce Parkinson's disease-like symptoms as well as neurotoxicity in humans and animal species. Recently, we reported that maintenance of redox balance and cellular defense against oxidative damage are primary functions of the novel antioxidant enzyme cytosolic NADP$^+$-dependent isocitrate dehydrogenase (IDPc). In this study, we examined the role of IDPc in cellular defense against MPP$^+$-induced oxidative injury using PC12 cells transfected with IDPc small interfering RNA (siRNA). Our results demonstrate that MPP$^+$-mediated disruption of cellular redox status, oxidative damage to cells, and apoptotic cell death were significantly enhanced by knockdown of IDPc.

Redescription of two soil ciliates, Anteholosticha bergeri and Bakuella granulifera, from South Korea

  • Chae, Kyu-Seok;Kim, Kang-San;Min, Gi-Sik
    • Journal of Species Research
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    • v.10 no.1
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    • pp.63-71
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    • 2021
  • Anteholosticha bergeri and Bakuella granulifera were isolated from soil samples collected from Muuidong and Songdo-dong, Incheon and confirmed new to South Korea. Including these two newly recorded species, 11 species of Anteholosticha and four species of Bakuella have been recorded in South Korea to date. Anteholosticha bergeri was discriminated from congeners by following characters: cortical granules, 12-16 macronuclei, 5-8 midventral pairs, 2-3 pretransverse cirri, 4-6 transverse cirri, and three dorsal kineties. Bakuella granulifera was identified by cortical granules, 5-11 buccal cirri, 2-5 frontoterminal cirri, 2-5 midventral cirri rows, and 8-12 transverse cirri. The Korean A. bergeri population corresponds to the Austrian population, except for the number of marginal and transverse cirri, and the Korean B. granulifera population corresponds to the Namibian population, except for body size. In addition, small subunit ribosomal RNA(18S rRNA) gene sequences from both species were determined.

Inhibition of Plasminogen Activator Inhibitor-1 Expression in Smoke-Exposed Alveolar Type II Epithelial Cells Attenuates Epithelial-Mesenchymal Transition

  • Song, Jeong-Sup;Kang, Chun-Mi
    • Tuberculosis and Respiratory Diseases
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    • v.70 no.6
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    • pp.462-473
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    • 2011
  • Background: Smoking is a risk factor for idiopathic pulmonary fibrosis (IPF), but the mechanism of the association remains obscure. There is evidence demonstrating that plasminogen activator inhibitor-1 (PAI-1) is involved in the progression of pulmonary fibrosis. This study was to determine whether the administration of small interfering RNA (siRNA) targeting PAI-1 or PAI-1 inhibitor to the cigarette smoking extract (CSE)-exposed rat alveolar type II epithelial cells (ATII cells) limits the epithelial-mesenchymal transition (EMT). Methods: ATII cells were isolated from lung of SD-rat using percoll gradient method and cultured with 5% CSE. The EMT was determined from the ATII cells by measuring the real-time RT PCR and western blotting after the PAI-1 siRNA transfection to the cells and after administration of tiplaxtinin, an inhibitor of PAI-1. The effect of PAI-1 inhibitor was also evaluated in the bleomycin-induced rats. Results: PAI-1 was overexpressed in the smoking exposed ATII cells and was directly associated with EMT. The EMT from the ATII cells was suppressed by PAI-1 siRNA transfection or administration of tiplaxtinin. Signaling pathways for EMT by smoking extract were through the phosphorylation of SMAD2 and ERK1/2, and finally Snail expression. Tiplaxtinin also suppressed the pulmonary fibrosis and PAI-1 expression in the bleomycin-induced rats. Conclusion: Our data shows that CSE induces rat ATII cells to undergo EMT by PAI-1 via SMAD2-ERK1/2-Snail activation. This suppression of EMT by PAI-1 siRNA transfection or PAI-1 inhibitor in primary type II alveolar epithelial cells might be involved in the attenuation of bleomycin-induced pulmonary fibrosis in rats.

Identification of Isoleucine-Accepting tRNA in Maize Mitochondria

  • Park, Young-In;Lee, Byung-Chul;Chang, Hyo-Ihl;Moon, A-Ree
    • BMB Reports
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    • v.28 no.6
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    • pp.494-498
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    • 1995
  • Maize mitochondrial tRNAs for isoleucine have been isolated using a putative $tRNA^{Ile}$ gene probe which has been previously isolated and characterized. It contains the 5'-CAT anticodon which would normally recognize the AUG methionine codon. The nucleotide sequence of one of these tRNAs has been partially determined, and contains a modified nucleotide at the first position of the anticodon. This type of posttranscriptional modification event could change the specificity of amino acid acceptance of a tRNA, unlike that deduced from the corresponding gene. An aminoacylation experiment also demonstrated that these purified tRNAs have isoleucine acceptance activity but no methionine-accepting activity.

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Application of Molecular Methods for the Identification of Acetic Acid Bacteria Isolated from Blueberries and Citrus Fruits

  • Gerard, Liliana Mabel;Davies, Cristina Veronica;Solda, Carina Alejandra;Corrado, Maria Belen;Fernandez, Maria Veronica
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.193-204
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    • 2020
  • Sixteen acetic acid bacteria (AAB) were isolated from blueberries and citric fruits of the Salto Grande region (Concordia, Entre Rios, Argentina) using enrichment techniques and plate isolation. Enrichment broths containing ethanol and acetic acid enabled maximum AAB recovery, since these components promote their growth. Biochemical tests allowed classification of the bacteria at genus level. PCR-RFLP of the 16S rRNA and PCR-RFLP of the 16S-23S rRNA intergenic spacer allowed further classification at the species level; this required treatment of the amplified products of 16S and 16S-23S ITS ribosomal genes with the following restriction enzymes: AluI, RsaI, HaeIII, MspI, TaqI, CfoI, and Tru9I. C7, C8, A80, A160, and A180 isolates were identified as Gluconobacter frateurii; C1, C2, C3, C4, C5, C6, A70, and A210 isolates as Acetobacter pasteurianus; A50 and A140 isolates as Acetobacter tropicalis; and C9 isolate as Acetobacter syzygii. The bacteria identified by 16S rRNA PCR-RFLP were validated by 16S-23S PCR-RFLP; however, the C1 isolate showed different restriction patterns during identification and validation. Partial sequencing of the 16S gene resolved the discrepancy.

Clinical Features and Associated Factors of Macrolide-Unresponsive Mycoplasma pneumonia and Efficacy Comparison Between Doxycycline, Tosufloxacin and Corticostreoid as a Second-Line Treatment (마크로라이드 불응성 마이코플라즈마 폐렴의 임상 양상 및 연관 인자와 2차 치료제로서 doxycycline, tosufloxacin 및 corticosteroid의 효능 비교)

  • Han Byeol Kang;Youngmin Ahn;Byung Wook Eun;Seungman Park
    • Pediatric Infection and Vaccine
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    • v.31 no.1
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    • pp.37-45
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    • 2024
  • Purpose: This study aimed to examine the clinical features and determinants of macrolide-unresponsive Mycoplasma pneumoniae pneumonia (MUMP) and to assess the differences in the time to fever resolution between doxycycline (DXC), tosufloxacin (TFX) and corticosteroid (CST) as second-line treatment. Methods: We retrospectively analyzed the medical records of patients under the age of 18 who were admitted to Nowon Eulji University Hospital between July 2018 and February 2020, diagnosed with mycoplasma pneumonia. Macrolide resistance was confirmed by detecting point mutations in the 23S rRNA gene. MUMP was clinically defined by persistent fever (≥38.0℃) lasting for 72 hours or more after the initiation of macrolide treatment. In cases of MUMP, patients were treated with an addition of CST, or the initial macrolide was replaced either DXC or TFX. Results: Out of 157 cases of mycoplasma pneumonia, 83 cases (52.9%) did not respond to macrolides. Patients with MUMP exhibited significantly higher C-reactive protein (CRP) levels (3.2±3.0 vs. 2.4±2.2 mg/dL, P=0.047), more frequent lobar/segmental infiltrations or pleural effusions (56.6% vs. 27.0%, P<0.001; 6.0% vs. 0.0%, P=0.032), and a higher prevalence of 23S rRNA gene mutations (96.4% vs. 64.6%, P<0.001) when compared to those with macrolide-susceptible M. pneumoniae pneumonia. In terms of second-line treatment, 15 patients (18.1%) responded to CST, 30 (36.1%) to DXC, and 38 (45.8%) to TFX. The time to defervescence (TTD) after initiation second-line treatment was significantly shorter in the CST group compared to the DXC (10.3±12.7 vs. 19.4±17.2 hours, P=0.003) and TFX groups (10.3±12.7 vs. 25.0±20.1 hours, P=0.043), with no significant difference observed between the DXC and TFX groups (19.4±17.2 vs. 25.0±20.1 hours, P=0.262). Conclusions: High CRP levels, the presence of positive 23S rRNA gene mutation, lobar or segmental lung infiltration, and pleural effusion observed in chest X-ray findings were significant factors associated with macrolide unresponsiveness. In this study, CST demonstrated a shorter TTD compared to DXC or TFX. Further, larger-scale prospective studies are needed to determine the optimal second-line treatment for MUMP.

Gene Expression of Surfactant Protein B and C in Endotoxin and Thiourea Treated Rats (내독소 및 Thiourea 투여 후 Surfactant protein B와 C 유전자 발현의 비교 관찰)

  • Sohn, Dong Hyun;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.54 no.5
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    • pp.510-521
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    • 2003
  • Background : The surfactant specific proteins, SP-B and SP-C are believed to be important regulators of the surfactant function and homeostasis. Since acute respiratory distress syndrome(ARDS) is usually viewed as the functional and morphological expression of a similar underlying lung injury caused by a variety of insults, and since abnormalities in the surfactant function have been described in ARDS, the authors investigated the different effects of endotoxin and thiourea on the accumulation of mRNA encoding SP-B and SP-C. Methods : Sprague-Dawley rats were given 5 mg/kg of an intraperitoneal endotoxin from Salmonella enteritidis and 3.5 mg/kg intraperitoneal thiourea and were sacrificed at different time periods. Results : 1. The SP-B mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 26.1% and 50%, respectively(P<0.01, P<0.001). 2. The SP-B mRNA levels 24 hours after the 3.5 mg/kg thiourea treatment was reduced by 9.8% and 12.5%, respectively. 3. The SP-C mRNA levels 6 and 24 hours after the 5 mg/kg endotoxin treatment was significantly reduced by 38.7% and 53.6%, respectively(P<0.01, P<0.001). 4. The SP-C mRNA level 6 hours after the 3.5 mg/kg thiourea treatment was reduced by 22.8%(P<0.05). Conclusion : These results indicate that the differential regulation of the hydrophobic surfactant proteins in vivo is evident, and suggest that the hydrophobic surfactant proteins might be differentially regulated during lung injury at different time periods without altering the lung wet to dry ratios. The mechanism of these alternations at the different time periods and the different kinds of etiology remain to be determined.