• Title/Summary/Keyword: small RNA

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Mapping, Tissue Distribution and Polymorphism of Porcine Retinol Binding Protein Genes (RBP5 and RBP7)

  • Gong, W.H.;Tang, Z.L.;Han, J.L.;Yang, S.L.;Wang, H.;Li, Y.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.11
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    • pp.1544-1550
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    • 2008
  • The retinoids (vitamin A and its derivatives) play a critical role in vision, growth, reproduction, cell differentiation and embryonic development. Using the IMpRH panel, porcine cellular retinol binding protein genes 5 and 7 (RBP5 and RBP7) were assigned to porcine chromosomes 5 and 6, respectively. The complete coding sequences (CDS) of the RBP5 and RBP7 genes were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) method, and the deduced amino acid sequences of both genes were compared to human corresponding proteins. The mRNA distributions of the two genes in adult Wuzhishan pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph node, small intestine, liver, brain, kidney and fat) were examined. A total of nine single nucleotide polymorphisms (SNPs) were identified in two genes. Three of these SNPs were analyzed using the polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) method in Laiwu, Wuzhishan, Guizhou, Bama, Tongcheng, Yorkshire and Landrace pig breeds. Association analysis of genotypes of these SNP loci with economic traits was done in our experimental populations. Significant associations of different genotypes of $RBP5-A/G^{63}$, $RBP5-A/G^{517}$ and $RPB5-T/C^{intron1-90}$ loci with traits including maximum carcass length (LM), minimum carcass length (LN), marbling score (MS), back fat thickness at shoulder (SBF), meat color score (MCS) and hematocrit (HCT) were detected. These SNPs may be useful as genetic markers in genetic improvement for porcine production.

Isolation of Enterovirus Causing Aseptic Meningitis in Busan, 2000-2002 Years (2000-2002년 바이러스성 뇌수막염의 발생양상 및 특성)

  • Jo, Gyeong-Sun;Jeong, Myeong-Ju
    • Korean Journal of Microbiology
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    • v.39 no.4
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    • pp.248-252
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    • 2003
  • Enteroviruses isolation were attempted from samples obtained from aseptic meningitis-suspected patients in hospitals in Busan during 2000-2002. Enteroviruses were found in 2 of 292 cases in 2000, 4 of 371 cases in 2001, 83 of 703 cases in 2002. In 2000, the isolated viruses were found to be echovirus serotype 11 and coxsackievirus serotype B2. Coxsackievirus serotype B5 was isolated in 2001 and in 2002, echovirus serotypes 2, 3, 6, 7, 9, 13, 25, 30 were isolated in 70 cases while coxsackievirus serotypes B3 and B4 were isolated in 10 cases. Various specimens tended to emerge over the years. The occurrence in 2000 tended to be mostly focus during the cold months, December through January, while in 2001, it occurred in May. In 2002, occurrence was found to be distributed from April to November with the highest rate during June and July. The strains of Vero and HEp-2 of echovirus and coxsackievirus, respectively, are highly infectious. Electron micrograph of echovirus and coxsackievirus show that they are small nonenevolped, isometric-shaped viruses. Isolated RNA from strains of echovirus and coxsackievirus showing cytopathic effects were used to undergo nested PCR which resulted in a 436 bp single band in all the strains. The serotype was sent to the Department of Virology at the Korean National Institute of Health for identification.

Expression of Heat Shock Protein and Antioxidant Genes in Rice Leaf Under Heat Stress

  • Lee, Dong-Gi;Ahsan, Nagib;Kim, Yong-Goo;Kim, Kyung-Hee;Lee, Sang-Hoon;Lee, Ki-Won;Rahman, Md. Atikur;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.3
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    • pp.159-166
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    • 2013
  • We have previously investigated the proteome changes of rice leaves under heat stress (Lee et al. in Proteomics 2007a, 7:3369-3383), wherein a group of antioxidant proteins and heat shock proteins (HSPs) were found to be regulated differently. The present study focuses on the biochemical changes and gene expression profiles of heat shock protein and antioxidant genes in rice leaves in response to heat stress ($42^{\circ}C$) during a wide range of exposure times. The results show that hydrogen peroxide and proline contents increased significantly, suggesting an oxidative burst and osmotic imbalance under heat stress. The mRNA levels of chaperone 60, HSP70, HSP100, chloroplastic HSP26, and mitochondrial small HSP responded rapidly and showed maximum expression after 0.5 or 2 h under heat stress. Transcript levels of ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR) and Cu-Zn superoxide dismutase (Cu-Zn SOD) showed a rapid and marked accumulation upon heat stress. While prolonged exposure to heat stress resulted in increased transcript levels of monodehydroascorbate reductase, peroxidase, glyoxalase 1, glutathione reductase, thioredoxin peroxidase, 2-Cysteine peroxiredoxin, and nucleoside diphosphate kinase 1, while the transcription of catalase was suppressed. Consistent with their changes in gene expression, the enzyme activities of APX and DHAR also increased significantly following exposure to heat stress. These results suggest that oxidative stress is usually caused by heat stress, and plants apply complex HSP- and antioxidant-mediated defense mechanisms to cope with heat stress.

Role of LPS-activated Macrophages in the Differentiation of Mesenchymal Stem Cells into Smooth Muscle Cells (중간엽 줄기세포의 평활근 세포로의 분화에서 LPS에 의해 활성화된 대식세포의 역할)

  • Lee, Mi Jeong;Do, Eun Kyoung;Kim, Jae Ho
    • Journal of Life Science
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    • v.23 no.1
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    • pp.137-142
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    • 2013
  • Human adipose-derived mesenchymal stem cells (hMSCs) are highly useful for vascular regeneration of injured or inflamed tissue. Lipopolysaccharide (LPS) is a potent activator of macrophages and stimulates macrophages to release inflammatory cytokines. In the present study, we explored the role of LPS-activated macrophages in the differentiation of hMSCs to smooth muscle cells (SMCs). We demonstrated that conditioned medium from LPS-induced macrophages (LPS CM) stimulates differentiation of hMSCs to SMCs, as evidenced by increased expression of smooth muscle-specific markers, including alpha-smooth muscle actin (${\alpha}$-SMA), smooth muscle-myosin heavy chain, and calponin. LPS induced the secretion of $PGF2{\alpha}$ from macrophages, and $PGF2{\alpha}$ treatment stimulated expression levels of SMC-specific markers in hMSCs. Furthermore, small interfering RNA-mediated silencing of the $PGF2{\alpha}$ receptor inhibited LPS CM-stimulated ${\alpha}$-SMA expression. These results suggest that LPS-activated macrophages promote differentiation of hMSCs to SMCs through a $PGF2{\alpha}$-dependent mechanism.

Fermentation of Cucurbita maxima Extracts with Microganisms from Kimchi (김치 유래 유산균을 이용한 단호박 발효음료 제조 기술 개발)

  • Roh, Hyun-Ji;Kim, Gi-Eun
    • KSBB Journal
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    • v.24 no.2
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    • pp.149-155
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    • 2009
  • 19 strains, which could be identified as Lactobacillus sp. were isolated. The Cucurbita maxima has been known as a traditional healthy food and variable positive effects on the human body were already reported. In this study we tried to develop a production process for a healthy fermented drink with Cucurbita maxima and strains originated from Kimchi. Many kinds of lacctobacci species existed in the fermented food cannot survive in the acidic conditions in the stomach. So we tried to search and select a strain, which can arrive to the small intestine. A species of a Lactobacillus named as C332 was identifed as Lactobacillus plantarum and selected for the fermentation process. With the treatment with artificial gastric juice and artificial bile the survival rate of the cells could be calculated. The physiological characteristics at the variable conditions have been tested. After fermentation process the sensoric tests on the product with panels were tried. The most of the cells could survive in the acidic conditions and falcultive anaerobe. Especially some antibacterial effects aganinst E.coli were also found. With all kinds of the results from our research the fermented Cucurbita maxima drink can be a successful item in the market.

Epigenetic Study of XIST Gene from Female and Male Cells by Pyrosequencing (남성과 여성에서 XIST 유전자의 후성학적 비교 연구)

  • Kim, Hwan-Hee;Yun, Yeo-Jin;Song, Min-Ae;Lee, Su-Man
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.1
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    • pp.25-31
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    • 2010
  • Objective: X inactivation is the silencing one of the two X chromosomes in female mammals for gene dosage on the X-chromosome between female and male. X inactivation is controlled by X inactive-specific transcript (XIST) gene, untranslated RNA. XIST is expressed only from the inactive X (Xi), not expressed from the active X (Xa). The Xist promoter is methylated on the silent Xist allele on the Xa in somatic cells, and less methylated on the Xist-expressing Xi. We investigated the difference of XIST methylation pattern of the promoter and 5'-region of XIST from male (XY) and female (XX) subjects. Methods: The direct quantification of XIST methylation is required for clinical application of normal XX and XY blood. Methylation percentage of eight CpG sites (-1696, -1679, -1475, -1473, -1469, +947, +956, +971) of XIST gene were diagnosed by pyrosequencing. Results: We directly quantitated the methylation percentage of the promoter and 5'-end of XIST by pyrosequencing. The average methylation percentages at CpG6-8 sites (+947, +956, +971) were 45.2% at CpG6, 49.9% at CpG7, and 44.2% at CpG8 from normal female and normal male were 90.6%, 96.7%, 87.8%, respectively. Nether CpG 1-5sites (-1696, -1679, -1475, -1473, -1469) had any effect on XX and XY. Conclusion: This method is sensitive for quantifying the small percentage change in the methylation status of XIST, and may be used for diagnosis.

Silybin Synergizes with Wnt3a in Activation of the Wnt/${\beta}$-catenin Signaling Pathway through Stabilization of Intracellular ${\beta}$-Catenin Protein (Silybin에 의한 Wnt/${\beta}$-catenin 신호전달체계의 활성화)

  • Kim, Tae-Yeoun;Oh, Sang-Taek
    • Microbiology and Biotechnology Letters
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    • v.40 no.1
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    • pp.50-56
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    • 2012
  • The Wnt/${\beta}$-catenin signaling pathway regulates diverse developmental processes and adult tissue homeostasis. Inappropriate regulation of this pathway has been associated with human diseases, such as cancers, osteoporosis, and Alzheimer's disease. Using a cell-based chemical screening with natural compounds, we discovered silybin, a plant flavonoid isolated from the Silybum marianum, which activated the Wnt/${\beta}$-catenin signaling pathway in a synergy with Wnt3a-conditioned medium (Wnt3a-CM). In the presence of Wnt3a-CM, silybin up-regulated ${\beta}$-catenin response transcription (CRT) in HEK293-FL reporter cells and 3T3-L1 preadipocytes through stabilization of intracellular ${\beta}$-catenin protein. Silybin and Wnt3a-CM synergistically reduced expression of important adipocyte marker genes including peroxisome-proliferator-activated $receptor{\gamma}$ ($PPAR{\gamma}$) and CAATT enhancer-binding protein ${\alpha}$ (C/$EBP{\alpha}$) in 3T3-L1 preadipocytes, accompanied by the activation of Wnt/${\beta}$-catenin signaling pathway. Taken together, our findings indicate that silybin is a small-molecule synergist of the Wnt/${\beta}$-catenin signaling pathway and can be used as a controllable reagent for investigating biological processes that involve the Wnt/${\beta}$-catenin signaling pathway.

FUN14 Domain-Containing Protein 1 Is Involved in Amyloid Beta Peptide-Induced Mitochondrial Dysfunction and Cell Injury in HT-22 Neuronal Cells (HT-22 신경세포에서 아밀로이드 베타 펩티드에 의한 미토콘드리아와 세포 손상 기전에서 FUN14 도메인 함유 단백 1의 역할)

  • Jae Hoon Kang;Jae Suk Woo
    • Journal of Life Science
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    • v.34 no.1
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    • pp.37-47
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    • 2024
  • FUN14 domain-containing protein 1 (FUNDC1), an outer mitochondrial membrane protein, contributes to removal of damaged mitochondria through mitophagy. In this study, to elucidate the role of the FUNDC1 in the amyloid beta peptide (Aβ)-induced neuropathy, changes in the degree of mitochondrial dysfunction and cell injury caused by Aβ treatment were examined in the HT-22 neuronal cells in which the FUNDC1 expression was transiently silenced or overexpressed. We found that Aβ treatment causes a time-dependent decrease of the FUNDC1 expression. In the Aβ-treated cells, there were a drop in MTT reduction ability, depletion of cellular ATP, disruption of mitochondrial membrane potential, stimulation of cellular ROS production, and increased mitochondrial Ca2+ load. Activation of caspase-3 and induction of apoptotic cell death were also observed. Transient silencing of the FUNDC1 expression by transfection with the FUNDC1 small interfering RNA per se caused mitochondrial dysfunction and apoptotic cell death like the effect of Aβ treatment. Conversely, in cells in which the FUNDC1 was transiently overexpressed by FUNDC1-Myc transfection, overexpression itself had no effect on the mitochondrial functional integrity and cell survival but showed a significant prevention effect against mitochondrial and cell injury caused by Aβ treatment. Overall, these results suggest that the FUNDC1 is importantly involved in the Aβ-induced mitochondrial dysfunction and cell injury in the HT-22 neuronal cells.

A Cyclin-Dependent Kinase Inhibitor, p16^{INK4A}, Induces Apoptosis in The Human Cancer Cells. (Cyclin-dependent Kinase저해 단백질 p16^{INK4A}의 인체 암세포에서의 세포사멸 유도 활성)

  • 김민경;이철훈
    • Microbiology and Biotechnology Letters
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    • v.32 no.1
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    • pp.72-77
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    • 2004
  • Previously, we synthesized a novel Cyclin-dependent kinase inhibitor, MCS-5A. Also, we investigated the involvement of cell cycle regulatory events during MCS-5A-mediated apoptosis in HL-60(+p16/-p53) cells with up-regulation of p16 protein expression. In contrast, apoptosis was not observed in A549(-p16/+p53) cells. Therefore we propose that $p16^{INK4A}$ is a key enzyme for inducing apoptosis. In the present studies, we have explored the mechanism of $p16^{INK4A}$ -mediated cytotoxicity and the role of p16.sup INK4A/ overexpression in the induction of apoptosis in human tumor cells. The tumor suppressor gene $p16^{INK4A}$ is known as a cyclin-dependent kinase inhibitor (CKI) and cell cycle regulator. We expressed wild type $p16^{INK4A}$ in pcDNA3.1 vector and then transfected into non-small cell lung cancer (NSCLC) cell expressing different statue of p16$^{INK4A}$, p53 gene〔A549(-p16/+p53), H1299(-p16/-p53) and HeLa(+pl6/+p53) cell line〕. TUNEL assay (including propidium iodide staining following transfection of these cell line with pcDNA3.1-pl6) indicate that p16$^{INK4A}$-mediated cytotoxicity was associated with apoptosis. This is supported by studies demonstrating an induction of caspase 3 cleavage due to the transfection of A549, H1299 and HeLa cells with pcDNA3.1-pl6. These results suggest that p16$^{INK4A}$ has a new function of inducing apoptosis which is not related with the function of tumor suppressor gene p53.

Telomerase Activity in Primary Lung Cancers (원발성 폐암에 있어서 Telomerase 활성도에 대한 연구)

  • Yun, Sang-Myung;Kwak, Kyung-Rok;Hwang, Jee-Yoon;Park, Sam-Seok;Jeon, Doo-Soo;Kim, Cheol-Min;Lee, Min-Ki;Park, Soon-Kew
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.2
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    • pp.195-203
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    • 1999
  • Background: Telomerase enzyme activity is not detected in most normal cells, a phonomenon believed to be associated with limitations on cellular proliferation. Since this activity is detected in nearly all human tumor, including lung cancers, it has been suggested that telomerase activation may be coupled to acquisition of malignant phenotype. In this study, we determined whether telomerase activity was associated with tumor pathologic stage. Methods: Primary tumor specimens obtained by bronchoscopic biopsies from 33 patients were analyzed. Telomerase activity was measured by means of a modified Telomeric Repeat Amplication Protocol(TRAP) assay. Results: Telomerase activity was detected in 23 of the 27 non-small-cell lung cancer and 5 of 6 small-cell lung cancer. A few primary tumors did not appear to have detectable telomerase activity. Positive associations were found between the telomerase-positive rate and tumor stage(p<0.05). Conclusion: High telomerase activity is detected frequently in primary lung cancers that exhibit high tumor cell proliferation rates and advanced pathologic stage.

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