• Title/Summary/Keyword: Gene regulation

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EFFECT OF CAPSAICIN AND EUGENOL ON ICGRP (IMMUNOREACTIVE CALCITONIN GENE-RELATED PEPTIDE) RELEASE FROM RAT LUMBAR SPINAL CORD. (백서 척수에서 Capsaicin과 Eugenol이 iCGRP (immunoreactive calcitonin gene-related peptide) 분비 조절에 미치는 영향.)

  • 오원만;김원재;최남기;박상원;황인남;김선헌
    • Restorative Dentistry and Endodontics
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    • v.26 no.5
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    • pp.436-442
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    • 2001
  • Neuropeptide such as calcitonin gene-related peptide and substance P may mediate neurogenic inflammation, but little is known about the regulation of neuropeptide release from rat spinal cord. Eugenol has been reported to reduce odontogenic pain and is known to have a structure similar to capsaicin, a potent stimulant of certain nociceptors. This study was done to examine the effect of capsaicin and eugenol on immunoreactive calcitonin gene-related peptide (iCGRP) release from rat spinal cord and whether eugenol regulates capsaicin-sensitive release of iCCRP or it evokes capsaicin-sensitive release of iCGRP. The dor-sal half of rat lumbar spinal cord was chopped into 200$\mu$m slices. They were superfused (500$\mu$l/min) in vitro with an oxygenated Kreb's buffer. The EC$_{50}$ of capsaicin on iCGRP release was measured. Eugenol (600$\mu$M and 1.2mM) and vehicle (0.02% 2-hydroxyl-$\beta$-cyclodextrin) were administered prior to stimulation of rat lumbar spinal cord with capsaicin. The amount of iCGRP release from rat lumbar spinal cord was measured by radioimmunoassay. The results were as follows : 1. iCGRP release from rat lumbar spinal cord was dependent on concentration of capsaicin. The EC$_{50}$ of capsaicin on iCGRP release was 3$\mu$M. 2. In the vehicle treated group, capsaicin (3$\mu$M) evoked a 14-fold increase over basal iCGRP level. 3. Administration of 600$\mu$M and 1.2mM eugenol evoked a 2.2-fold increase and a 2.3-fold increase over basal iCGRP level respectively. 4. Administration of 600$\mu$M and 1.2mM eugenol increased capsaicin evoked release of iCGRP by more than 50%. These results indicate that eugenol evoke CGRP release from central nervous system and potentiate the pain-inducing action of capsaicin on it.

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Bisphenol A Bis(2,3-dihydroxypropyl) ether (BADGE.2H2O) Induces Orphan Nuclear Receptor Nur77 Gene Expression and Increases Steroidogenesis in Mouse Testicular Leydig Cells

  • Ahn, Seung-Won;Nedumaran, Balachandar;Xie, Yuanbin;Kim, Don-Kyu;Kim, Yong Deuk;Choi, Hueng-Sik
    • Molecules and Cells
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    • v.26 no.1
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    • pp.74-80
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    • 2008
  • Bisphenol A bis (2,3-dihydroxypropyl) ether ($BADGE.2H_2O$) is a component of commercial liquid epoxy resins commonly used in the food-packing industry and in dental sealants. There is evidence that it has significant estrogenic activity. Nur77 plays a crucial role in the regulation of certain genes involved in LH-mediated steroidogenesis in testicular Leydig cells. It was previously demonstrated that Bisphenol A (BPA) stimulates Nur77 gene induction and steroidogenesis. In this study, we investigated the effects of $BADGE.2H_2O$ on Nur77 gene expression and steroidogenesis. Northern blot analysis showed that it increased the expression of Nur77 mRNA and protein, and transient transfection assays demonstrated that it increased the promoter activity and transactivation of Nur77. It also increased the expression of certain steroidogenic genes, such as StAR and $3{\beta}$-HSD. Finally, over-expression of a dominant negative Nur77 cDNA via adenoviral infection reduced $BADGE.2H_2O$-mediated progesterone biosynthesis. These results indicate that $BADGE.2H_2O$ disrupts testicular steroidogenesis by increasing Nur77 gene expression.

Divergence of Genes Encoding Non-specific Lipid Transfer Proteins in the Poaceae Family

  • Jang, Cheol Seong;Jung, Jae Hyeong;Yim, Won Cheol;Lee, Byung-Moo;Seo, Yong Weon;Kim, Wook
    • Molecules and Cells
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    • v.24 no.2
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    • pp.215-223
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    • 2007
  • The genes encoding non-specific lipid transfer proteins (nsLTPs), members of a small multigene family, show a complex pattern of expressional regulation, suggesting that some diversification may have resulted from changes in their expression after duplication. In this study, the evolution of nsLTP genes within the Poaceae family was characterized via a survey of the pseudogenes and unigenes encoding the nsLTP in rice pseudomolecules and the NCBI unigene database. nsLTP-rich regions were detected in the distal portions of rice chromosomes 11 and 12; these may have resulted from the most recent large segmental duplication in the rice genome. Two independent tandem duplications were shown to occur within the nsLTP-rich regions of rice. The genomic distribution of the nsLTP genes in the rice genome differs from that in wheat. This may be attributed to gene migration, chromosomal rearrangement, and/or differential gene loss. The genomic distribution pattern of nsLTP genes in the Poaceae family points to the existence of some differences among cereal nsLTP genes, all of which diverged from an ancient gene. The unigenes encoding nsLTPs in each cereal species are clustered into five groups. The somewhat different distribution of nsLTP-encoding EST clones between the groups across cereal species imply that independent duplication(s) followed by subfunctionalization (and/or neofunctionalization) of the nsLTP gene family in each species occurred during speciation.

Cloning and Expression Analysis of a Chitinase Gene Crchi1 from the Mycoparasitic Fungus Clonostachys rosea (syn. Gliocladium roseum)

  • Gan, Zhongwei;Yang, Jinkui;Tao, Nan;Yu, Zefen;Zhang, Ke-Qin
    • Journal of Microbiology
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    • v.45 no.5
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    • pp.422-430
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    • 2007
  • Clonostachys rosea (syn. Gliocladium roseum) is a well-known biocontrol agent and widely distributed around the world. In this study, an endochitinase gene Crchi1 was isolated from the mycoparasitic fungus C. rosea using the DNA walking strategy. The Crchi1 ORF is 1,746 bp long and interrupted by three introns. The cloned gene Crchi1 encodes 426 amino acid residues and shares a high degree of similarity with other chitinases from entomopathogenic and mycoparasitic fungi. Several putative binding sites for transcriptional regulation of Crchi1 in response to carbon (5'-SYGGRG-3') and nitrogen (5'-GATA-3') were identified in the upstream of Crchi1. Expression of Crchi1 gene in different carbon sources was analyzed using real-time PCR (RT-PCR). We found that the Crchi1 expression was suppressed by glucose but strongly stimulated by chitin or solubilized components of the cell wall from Rhizoctonia solani. Phylogenetic analysis of chitinases from entomopathogenic and mycoparasitic fungi suggests that these chitinases have probably evolved from a common ancestor.

Identification of CCL1 as a Gene Differentially Expressed in $CD4^+$ T cells Expressing TIM-3

  • Jun, Ka-Jung;Lee, Mi-Jin;Shin, Dong-Chul;Woo, Min-Yeong;Kim, Kyong-Min;Park, Sun
    • IMMUNE NETWORK
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    • v.11 no.4
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    • pp.203-209
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    • 2011
  • Background: T cell immunoglobulin mucin containing molecule (TIM)-3 is expressed in differentiated Th1 cells and is involved in the suppression of the cytokine production by these cells. However, the regulation of the expression of other T cell genes by TIM-3 is unclear. Herein, we attempted to identify differentially expressed genes in cells abundantly expressing TIM-3 compared to cells with low expression of TIM-3. Methods: TIM-3 overexpressing cell clones were established by transfection of Jurkat T cells with TIM-3 expression vector. For screening of differentially expressed genes, gene fishing technology based on reverse transcription-polymerase chain reaction (RT-PCR) using an annealing control primer system was used. The selected candidate genes were validated by semi quantitative and real-time RT-PCR. Results: The transcription of TIMP-1, IFITM1, PAR3 and CCL1 was different between TIM-3 overexpressing cells and control cells. However, only CCL1 transcription was significantly different in cells transiently transfected with TIM3 expression vector compared with control cells. CCL1 transcription was increased in primary human $CD4^+$ T cells abundantly expressing TIM-3 but not in cells with low expression of TIM-3. Conclusion: CCL1 was identified as a differentially transcribed gene in TIM-3-expressing $CD4^+$ T cells.

Implant surface treatments affect gene expression of Runx2, osteogenic key marker

  • Na, Young;Heo, Seong-Joo;Kim, Seong-Kyun;Koak, Jai-Young
    • The Journal of Advanced Prosthodontics
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    • v.1 no.2
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    • pp.91-96
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    • 2009
  • STATEMENT OF PROBLEM. The aim of this study was to study the effects of various surface treatments to a titanium surface on the expression of Runx2 in vitro. MATERIAL AND METHODS. Human Osteosarcoma TE-85 cells were cultured on machined, sandblasted, or anodic oxidized cpTi discs. At various times of incubation, the cells were collected and then processed for the analysis of mRNA expression of Runx2 using reverse transcription-PCR. RESULTS. The expression pattern of Runx2 mRNA was differed according to the types of surface treatment. When the cells were cultured on the untreated control culture plates, the gene expression of Runx2 was not increased during the experiments. In the case of that the cells were cultured on the machined cpTI discs, the expression level was intermediate at the first day, but increased constitutively to day 5. In cells on sandblasted cpTi discs, the expression level was highest in the first day sample and the level was maintained to 5 days. In cells on anodized cpTi discs, the expression level increased rapidly to 3 days, but decreased slightly in the 5-th day sample. CONCLUSION. Different surface treatments may contribute to the regulation of osteoblast function by influencing the level of gene expression of key osteogenic factors.

Association of the RORA Gene Polymorphism and Diurnal Preference in a Young Korean Population (RORA 유전자 다형성과 한국인 젊은 성인에서 일중선호도와의 연관성)

  • Ju, Hoon;Cho, Chul-Hyun;Moon, Joung Ho;Kim, Leen;Lee, Heon-Jeong
    • Sleep Medicine and Psychophysiology
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    • v.23 no.1
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    • pp.25-28
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    • 2016
  • Objectives: The retinoid-related orphan receptor A (RORA) gene has been reported to have an impact on circadian rhythm regulation. In this study, we analyzed the relationship between the RORA gene polymorphism and diurnal preference in Korean young adults. Methods: A population of 504 young adults was included in the study. All subjects were given and completed a 13-item composite scale for morningness (CSM). The RORA gene rs11071547 single-nucleotide polymorphism (SNP) was genotyped by PCR-based methods. Results: CSM score was not associated with genotype or allele carrier status of the RORA rs11071547 SNP. Conclusion: This result indicates that the RORA rs11071547 SNP does not play a role in diurnal preference.

CACNA1A Gene Polymorphism is Associated with Hypertension in Korean Population

  • Kim, Hye-Kyung;Leem, Kang-Hyun;Lee, Se-Na;Hong, Mee-Sook;Jung, Kyung-Hee;Kim, Su-Kang;Kang, Sung-Wook;Chung, Joo-Ho
    • Biomedical Science Letters
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    • v.15 no.1
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    • pp.31-35
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    • 2009
  • High blood pressure (BP) is the most frequent risk factor among metabolic syndrome components. The control of hypertension is very important to prevent the cardiovascular risk in metabolic syndrome. The dysfunction of calcium channel is responsible in the regulation of the vascular muscle contribution to hypertension. Calcium channel, voltage-dependent, P/Q type, alpha-1A subunit (CACNA1A) gene is located in brain and known to control the intracranial hypertension. In this study, we investigate whether the polymorphisms of CACNA1A gene is associated with hypertension. The 49 CACNA1A genotypes were determined using the Affymetrix Genotyping chip array in 92 hypertension and 279 control individuals from a Korean population. Logistic and multiple regression models were employed to analyze the genetic contributions of polymorphisms. Out of 49 polymorphisms, six SNPs (rs12611029, rs16035, rs7259944, rs10419472, rs17777900, and rs4926294) showed a significant association with hypertension in three alternative models (codominant, dominant, and recessive models; P<0.05 after adjusting for age and sex). Our results suggest that the CACNA1A gene may be associated with hypertension in the Korean population.

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NF-κB-dependent Regulation of Matrix Metalloproteinase-9 Gene Expression by Lipopolysaccharide in a Macrophage Cell Line RAW 264.7

  • Rhee, Jae-Won;Lee, Keun-Wook;Kim, Dong-Bum;Lee, Young-Hee;Jeon, Ok-Hee;Kwon, Hyung-Joo;Kim, Doo-Sik
    • BMB Reports
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    • v.40 no.1
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    • pp.88-94
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    • 2007
  • Matrix metalloproteinase-9 (MMP-9) plays a pivotal role in the turnover of extracellular matrix (ECM) and in the migration of normal and tumor cells in response to normal physiologic and numerous pathologic conditions. Here, we show that the transcription of the MMP-9 gene is induced by lipopolysaccharide (LPS) stimulation in cells of a macrophage lineage (RAW 264.7 cells). We provide evidence that the NF-$\kappa$B binding site of the MMP-9 gene contributes to its expression in the LPS-signaling pathway, since mutation of NF-$\kappa$B binding site of MMP-9 promoter leads to a dramatic reduction in MMP-9 promoter activation. In addition, the degradation of l$\kappa$B$\alpha$;, and the presences of myeloid differentiation protein (MyD88) and tumor necrosis factor receptor-associated kinase 6 (TRAF6) were found to be required for LPS-activated MMP-9 expression. Chromatin immunoprecipitation (ChIP) assays showed that functional interaction between NF-$\kappa$B and the MMP-9 promoter element is necessary for LPS-activated MMP-9 induction in RAW 264.7 cells. In conclusion, our observations demonstrate that NF-$\kappa$B contributes to LPS-induced MMP-9 gene expression in a mouse macrophage cell line.

Overexpression of Heat Shock Factor Gene HsfA3 Increases Galactinol Levels and Oxidative Stress Tolerance in Arabidopsis

  • Song, Chieun;Chung, Woo Sik;Lim, Chae Oh
    • Molecules and Cells
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    • v.39 no.6
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    • pp.477-483
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    • 2016
  • Heat shock factors (Hsfs) are central regulators of abiotic stress responses, especially heat stress responses, in plants. In the current study, we characterized the activity of the Hsf gene HsfA3 in Arabidopsis under oxidative stress conditions. HsfA3 transcription in seedlings was induced by reactive oxygen species (ROS), exogenous hydrogen peroxide ($H_2O_2$), and an endogenous $H_2O_2$ propagator, 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB). HsfA3-overexpressing transgenic plants exhibited increased oxidative stress tolerance compared to untransformed wild-type plants (WT), as revealed by changes in fresh weight, chlorophyll fluorescence, and ion leakage under light conditions. The expression of several genes encoding galactinol synthase (GolS), a key enzyme in the biosynthesis of raffinose family oligosaccharides (RFOs), which function as antioxidants in plant cells, was induced in HsfA3 overexpressors. In addition, galactinol levels were higher in HsfA3 overexpressors than in WT under unstressed conditions. In transient transactivation assays using Arabidopsis leaf protoplasts, HsfA3 activated the transcription of a reporter gene driven by the GolS1 or GolS2 promoter. Electrophoretic mobility shift assays showed that GolS1 and GolS2 are directly regulated by HsfA3. Taken together, these findings provide evidence that GolS1 and GolS2 are directly regulated by HsfA3 and that GolS enzymes play an important role in improving oxidative stress tolerance by increasing galactinol biosynthesis in Arabidopsis.