• Title/Summary/Keyword: 6S RNA

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The Effects of Rhus Extracts on The Cytotoxicity on Cancer Cells and E6 and E7 Oncogenes of Human Papillomavirus Type 16 (옻 추출물의 세포독성 및 자궁 경부암 바이러스 암 유발인자 E6 와 E7의 작용에 미치는 효과)

  • Cho, Young-Sik;Joung, Ok;Cho, Cheong-Weon;Lee, Kyung-Ae;Shim, Jung-Hyun;Kim, Kwang-Soo;Lee, Hong-Soo;Seung, Ki-Seung;Yoon, Do-Young
    • Korean Journal of Food Science and Technology
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    • v.32 no.6
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    • pp.1389-1395
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    • 2000
  • Cervical cancer has been one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papillomaviruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, Rhus, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Rhus inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, Caski) and HaCaT keratinocytes in a dose response manner. In vitro binding assay and ELISA showed that Rhus inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Rhus inhibited the in vitro binding of E7 and Rb which essential tumor suppressor for the control of cell cycle. The level of mRNA for E6 was also decreased by Rhus while that of E7 mRNA was not changed. Our data suggested that Rhus inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas by HPV.

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Characterization of Protoplast Fusant between Killer Yeast and Alcohol-Fermenting Yeast (Killer 효모와 알콜 발효효모간의 원형질체 융합주의 특성)

  • 정기택;방광웅;김재근;송형익;정용진
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.55-64
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    • 1990
  • Cell volume and DNA contents of the fusants were similar to those of parents. Genetic stability of the fusants was increased when they were cultured on minimal medium (MM) rather than on complete medium (CM), and the fusants were stabilized by subculturing 7 generations each 7 day on MM agar. The finally selected fusants after being cultured for 6 months on CM were stable without segregation. The fusants could also form nuclein and ascospores, and show red and pink colors by the test of TTC colorization. Assimilability and fermentability of carbon sources of the fusants were similarto those of parents. The tolerance of KCl, NaCl, sodium propionate and cycloheximide showed the traits of one strain of parents. When the fusants were cultured for 72 hr and 60 hr in the medium containing 20% glucose and sucrose, respectively, the yield of ethanol for FWKS 260 was reached to 9.6 v/v% and 9.8v/v%, respectively. The sensitive strain Kyokai 7 was found to be killed entirely after cultivation of 48 hr by the killer toxin from the fusants. The recipient S 29 and Kyokai 7 were found to have neither L nor M dsRNA plasmid. However, K 52 and fusants had both L and M dsRNA plasmid of 4.7 kb and 2.5 kb, respectively. The curants treated by heat and cycloheximide did not contain M dsRNA plasmid, but had large amounts of L dsRNA plasmid of those of killer yeasts.

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The Effects of NogYongDaeBoTang,(NYDBT)on the Alzheimer's Disease Model Induced by CT-105 and $A{\beta}$ (녹용대보탕이 ${\beta}-Amyloid$로 유도(誘導)된 Alzheimer's Disease 병태(病態) 모델에 미치는 영향(影響))

  • Seo, Gyoo-Tae;Lee, Eun-Kyung;Choi, Cheol-Hong;Chung, Dae-Kyoo
    • Journal of Oriental Neuropsychiatry
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    • v.18 no.2
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    • pp.101-132
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    • 2007
  • Objective : This research investigates the effect of the NogYongDaeBoTang,(NYDBT) on Alzheimer's disease. Method : The effects of the NYDBT extract on (1) $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ mRNA of PC-12 cells treated with LPS; (2) acetylcholinesterase(AChE), amyloid precursor proteins(APP), and glial fibrillary acidic protein(GFAP) mRNA the AChE activity and the APP production of PC-12 cell treated with CT-105; (3) the behavior; (4) expression of $IL-1{\beta}$, $TNF-{\alpha}$, MDA, $IL-1{\beta}$ mRNA, and $TNF-{\alpha}$ mRNA; (5) the infarction area of the hippocampus, and brain tissue injury in Alzheimer‘s diseased mice induced with ${\beta}A$ were investigated. Results : 1. The NYDBT extract suppressed the expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNA in BV2 microglia cell line treated with LPS. 2. The NYDBT extract suppressed the expression of $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ protein production in BV2 microglia cell line treated with LPS. 3. For the NYDBT extract group a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by $A{\beta}$ in the Morris water maze experiment, which measured stop-through latency, and distance movement-through latency. 4. The NYDBT extract suppressed the over-expression of $IL-1{\beta}$ protein, $TNF-{\alpha}$ protein, MDA, and CD68/CD11b, in the mice with Alzheimer's disease induced by $A{\beta}$. 5. The NYDBT extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by $A{\beta}$. 6. The NYDBT extract reduced the Tau protein, GFAP protein, and presenilin1/2 protein (immunohistochemistry) of hippocampus in the mice with Alzheimer's disease induced by $A{\beta}$. Conclusions : These results suggest that the NYDBT extract may be effective for the prevention and treatment of Alzheimer's disease.

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Monitoring of Chicken RNA Integrity as a Function of Prolonged Postmortem Duration

  • Malila, Yuwares;Srimarut, Yanee;U-chupaj, Juthawut;Strasburg, Gale;Visessanguan, Wonnop
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.11
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    • pp.1649-1656
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    • 2015
  • Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.

Bacterial diversity in children's dental caries (소아의 치아 우식 부위별 세균 다양성)

  • Kim, Eun-Mi;Baik, Keun-Sik;Ha, Myung-Ok
    • Journal of Korean society of Dental Hygiene
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    • v.13 no.5
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    • pp.889-900
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    • 2013
  • Objectives : Molecular biology techniques were employed to assess diversity of bacterial in children's dental caries. Methods : DNA of germs was extracted and the diversity of the 16S rRNA clones was analyzed by amplified rDNA restriction analysis and sequencing. The experimental samples were pit and fissure caries (PC), deep dentinal caries (DC), smooth surface caries (SC), and supragingival plaque (PQ) from 50 children of age less than 12 years old. The control group was healthy teeth supragingival plaque (HT). Thirty clones from each 16S rRNA clone library of 5 samples were randomly selected, thus a total of 150 clones were analyzed. Results : Amplified rDNA restriction analysis uncovered 18, 20, 11, 17, and 22 phylotypes from healthy teeth, pit and fissure caries, deep dentinal caries, smooth surface caries, and supragingival plaque, respectively. Sequencing analysis found the dominance of Actinomycs naeslundii and Fusobacterium nucleatum in the healthy teeth; Leptotrichia sp. in the pit and fissure caries; Actinomyces sp., Streptococcus mutans, and Rahnella aquatilis in the deep dentinal caries; Streptococcus mutans and Actinomyces sp. in the smooth surface caries; Enterobacter hormaechei and Streptococcus sanguinis in the supragingival plaque. Conclusions : Clonal analysis identified 6 phyla, 20 genera, and 51 species.

Archaeal Communities in Mangrove Soil Characterized by 16S rRNA Gene Clones

  • Yan, Bing;Hong, Kui;Yu, Zi-Niu
    • Journal of Microbiology
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    • v.44 no.5
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    • pp.566-571
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    • 2006
  • An archaeal 16S rRNA gene library was constructed from mangrove soil. Phylogenetic analysis revealed archaea in mangrove soil including the Crenarchaeota (80.4%) and Euryarchaeota (19.6%) phyla. The archaeal community in mangrove soil appears to be a mixture of organisms found in a variety of environments with the majority being of marine origin.

Roles of miR-128 in Myogenic Differentiation and Insulin Signaling in Rat L6 Myoblasts (쥐L6 근원세포에서 miR-128의 근육세포 분화와 인슐린신호에서의 역할)

  • Oh, Myung-Ju;Kim, So-Hyeon;Kim, Ji-Hyun;Jhun, Byung H.
    • Journal of Life Science
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    • v.30 no.9
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    • pp.772-782
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    • 2020
  • Skeletal muscle differentiation or myogenesis is important to maintain muscle mass and metabolic homeostasis. Muscle-specific microRNAs (miRNAs) are known to play a critical role in skeletal myogenic differentiation. In this study, we examined the expression profiling of miRNAs during myogenic differentiation in rat L6 myoblasts using rat miRNA microarrays. We identified the upregulated expression of miR-128 as well as several well-known myogenic miRNAs, including miR-1, miR-133b, and miR-206. We additionally confirmed the increased expression of miR-128 observed on microarray through quantitative real-time PCR (qRT-PCR), which showed similarly upregulated expression of both primary miR-128 and mature miR-128, consistent with the microarray findings. Furthermore, transfection of miR-128 into rat L6 myoblasts induced gene expression of myogenic markers such as muscle creatine kinase (MCK), myogenin, and myosin heavy chain (MHC). Protein expression of MHC was increased as well. Inhibition of miR-128 by inhibitory peptide nucleic acids (PNAs) blocked the expression of those myogenic markers. In addition, the transfection of miR-128 into rat L6 myoblasts enhanced the phosphorylation of Erk and Akt proteins stimulated by insulin, while simultaneously reversing the inhibited phosphorylation of Erk and Akt due to insulin resistance. These findings suggest that miR-128 may play important roles in myogenic differentiation and insulin signaling.

Microbial Community Structure in Hexadecane- and Naphthalene-Enriched Gas Station Soil

  • Baek, Kyung-Hwa;Kim, Hee-Sik
    • Journal of Microbiology and Biotechnology
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    • v.19 no.7
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    • pp.651-657
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    • 2009
  • Shifts in the activity and diversity of microbes involved in aliphatic and aromatic hydrocarbon degradation in contaminated soil were investigated. Subsurface soil was collected from a gas station that had been abandoned since 1995 owing to ground subsidence. The total petroleum hydrocarbon content of the sample was approximately 2,100 mg/kg, and that of the soil below a gas pump was over 23,000 mg/kg. Enrichment cultures were grown in mineral medium that contained hexadecane (H) or naphthalene (N) at a concentration of 200 mg/l. In the Henrichment culture, a real-time PCR assay revealed that the 16S rRNA gene copy number increased from $1.2{\times}10^5$to $8.6{\times}10^6$with no lag phase, representing an approximately 70-fold increase. In the N-enrichment culture, the 16S rRNA copy number increased about 13-fold after 48 h, from $6.3{\times}10^4$to $8.3{\times}10^5$. Microbial communities in the enrichment cultures were studied by denaturing gradient gel electrophoresis and by analysis of 16S rRNA gene libraries. Before the addition of hydrocarbons, the gas station soil contained primarily Alpha- and Gammaproteobacteria. During growth in the H-enrichment culture, the contribution of Bacteriodetes to the microbial community increased significantly. On the other hand, during N-enrichment, the Betaproteobacteria population increased conspicuously. These results suggest that specific phylotypes of bacteria were associated with the degradation of each hydrocarbon.

The Molecular Mechanism of Long Non-Coding RNA MALAT1-Mediated Regulation of Chondrocyte Pyroptosis in Ankylosing Spondylitis

  • Chen, Wei;Wang, Feilong;Wang, Jiangtao;Chen, Fuyu;Chen, Ting
    • Molecules and Cells
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    • v.45 no.6
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    • pp.365-375
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    • 2022
  • Long non-coding RNAs (lncRNAs) may be important regulators in the progression of ankylosing spondylitis (AS). The competing endogenous RNA (ceRNA) activity of lncRNAs plays crucial roles in osteogenesis. We identified the mechanism of the differentially expressed lncRNA MALAT1 in AS using bioinformatic analysis and its ceRNA mechanism. The interaction of MALAT1, microRNA-558, and GSDMD was identified using integrated bioinformatics analysis and validated. Loss- and gain-of-function assays evaluated their effects on the viability, apoptosis, pyroptosis and inflammation of chondrocytes in AS. We found elevated MALAT1 and GSDMD but reduced miR-558 in AS cartilage tissues and chondrocytes. MALAT1 contributed to the suppression of cell viability and facilitated apoptosis and pyroptosis in AS chondrocytes. GSDMD was a potential target gene of miR-558. Depletion of MALAT1 expression elevated miR-558 by inhibiting GSDMD to enhance cell viability and inhibit inflammation, apoptosis and pyroptosis of chondrocytes in AS. In summary, our key findings demonstrated that knockdown of MALAT1 served as a potential suppressor of AS by upregulating miR-558 via the downregulation of GSDMD expression.

Biosenesis of Epstein-Barr Virus MicroRNAs in B Cells (B 세포에서 Epstein-Barr virus microRNA들의 전사 및 성숙)

  • Kim Do Nyun;Oh Sang Taek;Lee Jae Myun;Lee Won-Keun;Lee Suk Kyeong
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.909-915
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    • 2005
  • We investigated microRNA (miRNA) biogenesis of Epstein-Barr virus (EBV) which is the first virus shown to produce viral miRNAs. As expected, expression of all the reported EBV miRNAs were detected by Northen blot in an EBV-infected B cell line, B95-8; BHRF1-1, BHIU1-2, BHRF1-3, BART1, and BART2. The putative EBV pri-miRWAs and pre-miRNAs predicted from the known mature EBV miRNA sequences were detected by RT-PCR in B95-8 cells. Many animal miRNA genes exist as clusters of 2-7 genes and they are expressed polycistronically. As the EBV miRNAs are clustered in two regions of the EBV genome, we examined whether these clustered EBV miRNA genes are also expressed polycistronically. A long polycistronic transcript with the expected size (1602 bp) corresponding to the BHRF1-1~BHRF1-2~BHRF1-3 was amplified. However, any polycistronic transcript containing both BART1 and BART2 was detectable in B95-8. These results suggest that EBV miRNAs may be processed in a similar way with animal miRNAs and that some of the clustered EBV miRNAs can be transcribed polycistronically.