• Molecules and Cells
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• 제40권7호
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• pp.476-484
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• 2017

C-X-C chemokine receptor 4 (CXCR4) stimulates cancer metastasis. NF-${\kappa}B$ regulates CXCR4 expression in cancer cells, and O-GlcNAc modification of NF-${\kappa}B$ promotes its transcriptional activity. Here, we determined whether CXCR4 expression is affected by O-GlcNAcylation of NF-${\kappa}B$ in lung metastasis of cervical cancer. We found elevated levels of O-linked-N-actylglucosamine transferase (OGT) and O-GlcNAcylation in cervical cancer cells compared to those in non-malignant epithelial cells and detected increased expression of NF-${\kappa}B$ p65 (p65) and CXCR4 in cervical cancer cells. Knockdown of OGT inhibited the O-GlcNAcylation of p65 and decreased CXCR4 expression levels in HeLa cells. Thiamet G treatment increased O-GlcNAcylated p65, which subsequently enhanced CXCR4 expression levels. Inhibition of O-GlcNAcylation by 6-Diazo-5-oxo-L-norleucine (DON) treatment decreased p65 activation, eventually inhibiting CXCR4 expression in HeLa cells. Lung tissues from mice engrafted with OGT-knockdown HeLa cells (shOGT) exhibited lower expression of Ki-67 and HPV E6 and E7 oncogenes compared to lung tissues from mice engrafted with control HeLa cells (shCTL). In addition, lung tissues from mice engrafted with shOGT cells exhibited lower p65 and CXCR4 immunoreactivity compared to tissues from mice engrafted with shCTL cells. Taken together, our data suggest that p65 O-GlcNAcylation promotes lung metastasis of cervical cancer cells by activating CXCR4 expression.

• Journal of Genetic Medicine
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• 제7권1호
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• pp.37-44
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• 2010

목적: Kabuki 증후군은 정신 지체를 동반하는 선천성 다발성 기형 증후군이다. 우리나라에서는 현재까지 6례의 Kabuki 증후군 증례가 산발적으로 보고 된 바 있다. 본 연구에서는 저자들이 경험한 Kabuki 증후군 환자 6명의 임상 및 유전학적 특징을 조사하고, 이를 외국 문헌들과 비교 분석해 보고자 하였다. 대상 및 방법: 2003년부터 2009년까지 아주대학교 병원 유전질환 전문센터에서 Kabuki 증후군으로 진단되어 추적 중인 6명의 한국인 여아를 대상으로 하였으며, 의무기록을 후향적으로 검토하여 이들의 임상 및 검사 소견을 수집하고 분석하였다. 결 과: 6명의 환자 모두가 특징적인 얼굴 모습 및 발달 지연 소견을 보이고 있었고, 손끝의 태아 패드 또한 모든 환자에서 확인되었다. 이외에도 대부분의 환자가 생후 성장 지연(83.3 %) 및 근력 저하(83.3%) 소견을 보였다. 안과적인 이상 또한 흔하게 동반되었는데, 특히 사시(83.3%)가 가장 흔한 안과적 이상 소견이었다. 선천성 심장 기형은 50%의 환자에서 동반되었으며, 골격계통의 증상으로는 짧거나(83.3%) 굽은(50%) 5번째 손가락, 관절의 과신전(50%) 및 고관절 탈구(16.7%) 등으로 다양하였다. Kabuki 증후군의 가족력을 가진 환자는 없었으며, 핵형 분석 및 array CGH를 포함한 세포유전학적 분석에서 Kabuki 증후군의 원인으로 생각되는 이상 소견은 발견되지 않았다. 결 론: 한국인 Kabuki 증후군 환자들이 보이는 임상 양상은 매우 광범위하며 다양한 신체 기관을 침범하고 있다. 비록 Kabuki 증후군의 임상적특징들이 비교적잘 알려져 있기는 하지만, 아직질환의 원인으로 추정되는 유전적 이상은 확실히밝혀지지 않았다. 적절한 질환의 관리및 유전상담이 이루어지기 위해서는 Kabuki 증후군의 자연 경과 및 유전적 배경에 대한 추가적인 연구가 필요하다고 생각된다.

• Journal of Animal Science and Technology
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• 제57권5호
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• pp.18.1-18.8
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• 2015

Background: Quantitative real time reverse transcription PCR (qRT-PCR) is one of the most important techniques for gene-expression analysis in molecular based studies. Selecting a proper internal control gene for normalizing data is a crucial step in gene expression analysis via this method. The expression levels of reference genes should be remained constant among cells in different tissues. However, it seems that the location of cells in different tissues might influence their expression. The purpose of this study was to determine whether the source of mesenchymal stem cells (MSCs) has any effect on expression level of three common reference genes (GAPDH, ${\beta}$-actin and ${\beta}2$-microglobulin) in equine marrow- and adipose-derived undifferentiated MSCs and consequently their reliability for comparative qRT-PCR. Materials and methods: Adipose tissue (AT) and bone marrow (BM) samples were harvested from 3 mares. MSCs were isolated and cultured until passage 3 (P3). Total RNA of P3 cells was extracted for cDNA synthesis. The generated cDNAs were analyzed by quantitative real-time PCR. The PCR reactions were ended with a melting curve analysis to verify the specificity of amplicon. Results: The expression levels of GAPDH were significantly different between AT- and BM-derived MSCs (p < 0.05). Differences in expression level of ${\beta}$-actin (P < 0.001) and B2M (P < 0.006.) between MSCs derived from AT and BM were substantially higher than GAPDH. In addition, the fold change in expression levels of GAPDH, ${\beta}$-actin and B2M in AT-derived MSCs compared to BM-derived MSCs were 2.38, 6.76 and 7.76, respectively. Conclusion: This study demonstrated that GAPDH and especially ${\beta}$-actin and B2M express in different levels in equine AT- and BM-derived MSCs. Thus they cannot be considered as reliable reference genes for comparative quantitative gene expression analysis in MSCs derived from equine bone marrow and adipose tissue.

• The Korean Journal of Physiology and Pharmacology
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• 제16권6호
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• pp.413-422
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• 2012

The purpose of this study is to investigated the mechanism of pharmacological action of local anesthetic and provide the basic information about the development of new effective local anesthetics. Fluorescent probe techniques were used to evaluate the effect of lidocaine HCl on the physical properties (transbilayer asymmetric lateral and rotational mobility, annular lipid fluidity and protein distribution) of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex, and liposomes of total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from the SPMV. An experimental procedure was used based on selective quenching of 1,3-di(1-pyrenyl)propane (Py-3-Py) and 1,6-diphenyl-1,3,5-hexatriene (DPH) by trinitrophenyl groups, and radiationless energy transfer from the tryptophans of membrane proteins to Py-3-Py. Lidocaine HCl increased the bulk lateral and rotational mobility of neuronal and model membrane lipid bilayes, and had a greater fluidizing effect on the inner monolayer than the outer monolayer. Lidocaine HCl increased annular lipid fluidity in SPMV lipid bilayers. It also caused membrane proteins to cluster. The most important finding of this study is that there is far greater increase in annular lipid fluidity than that in lateral and rotational mobilities by lidocaine HCl. Lidocaine HCl alters the stereo or dynamics of the proteins in the lipid bilayers by combining with lipids, especially with the annular lipids. In conclusion, the present data suggest that lidocaine, in addition to its direct interaction with proteins, concurrently interacts with membrane lipids, fluidizing the membrane, and thus inducing conformational changes of proteins known to be intimately associated with membrane lipid.

• The Korean Journal of Physiology and Pharmacology
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• 제22권3호
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• pp.311-319
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• 2018

Mitochondrial calcium overload is a crucial event in determining the fate of neuronal cell survival and death, implicated in pathogenesis of neurodegenerative diseases. One of the driving forces of calcium influx into mitochondria is mitochondria membrane potential (${\Delta}{\psi}_m$). Therefore, pharmacological manipulation of ${\Delta}{\psi}_m$ can be a promising strategy to prevent neuronal cell death against brain insults. Based on these issues, we investigated here whether nobiletin, a Citrus polymethoxylated flavone, prevents neurotoxic neuronal calcium overload and cell death via regulating basal ${\Delta}{\psi}_m$ against neuronal insult in primary cortical neurons and pure brain mitochondria isolated from rat cortices. Results demonstrated that nobiletin treatment significantly increased cell viability against glutamate toxicity ($100{\mu}M$, 20 min) in primary cortical neurons. Real-time imaging-based fluorometry data reveal that nobiletin evokes partial mitochondrial depolarization in these neurons. Nobiletin markedly attenuated mitochondrial calcium overload and reactive oxygen species (ROS) generation in glutamate ($100{\mu}M$)-stimulated cortical neurons and isolated pure mitochondria exposed to high concentration of $Ca^{2+}$ ($5{\mu}M$). Nobiletin-induced partial mitochondrial depolarization in intact neurons was confirmed in isolated brain mitochondria using a fluorescence microplate reader. Nobiletin effects on basal ${\Delta}{\psi}_m$ were completely abolished in $K^+-free$ medium on pure isolated mitochondria. Taken together, results demonstrate that $K^+$ influx into mitochondria is critically involved in partial mitochondrial depolarization-related neuroprotective effect of nobiletin. Nobiletin-induced mitochondrial $K^+$ influx is probably mediated, at least in part, by activation of mitochondrial $K^+$ channels. However, further detailed studies should be conducted to determine exact molecular targets of nobiletin in mitochondria.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2000년도 제18회 학술발표회 논문개요집
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• pp.77-77
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• 2000

Titanium oxide (TiO2) thin films have valuable properties such as a high refractive index, excellent transmittance in the visible and near-IR frequency, and high chemical stability. Therefore it is extensively used in anti-reflection coating, sensor, and photocatalysis as electrical and optical applications. Specially, TiO2 have a high dielectric constant of 180 along the c axis and 90 along the a axis, so it is highlighted in fabricating dielectric capacitors in micro electronic devices. A variety of methods have been used to produce patterned self-assembled monolayers (SAMs), including microcontact printing ($\mu$CP), UV-photolithotgraphy, e-beam lithography, scanned-probe based micro-machining, and atom-lithography. Above all, thin film fabrication on $\mu$CP modified surface is a potentially low-cost, high-throughput method, because it does not require expensive photolithographic equipment, and it produce micrometer scale patterns in thin film materials. The patterned SAMs were used as thin resists, to transfer patterns onto thin films either by chemical etching or by selective deposition. In this study, we deposited TiO2 thin films on Si (1000 substrateds using titanium (IV) isopropoxide ([Ti(O(C3H7)4)] ; TIP as a single molecular precursor at deposition temperature in the range of 300-$700^{\circ}C$ without any carrier and bubbler gas. Crack-free, highly oriented TiO2 polycrystalline thin films with anatase phase and stoichimetric ratio of Ti and O were successfully deposited on Si(100) at temperature as low as 50$0^{\circ}C$. XRD and TED data showed that below 50$0^{\circ}C$, the TiO2 thin films were dominantly grown on Si(100) surfaces in the [211] direction, whereas with increasing the deposition temperature to $700^{\circ}C$, the main films growth direction was changed to be [200]. Two distinct growth behaviors were observed from the Arhenius plots. In addition to deposition of THe TiO2 thin films on Si(100) substrates, patterning of TiO2 thin films was also performed at grown temperature in the range of 300-50$0^{\circ}C$ by MOCVD onto the Si(100) substrates of which surface was modified by organic thin film template. The organic thin film of SAm is obtained by the $\mu$CP method. Alpha-step profile and optical microscope images showed that the boundaries between SAMs areas and selectively deposited TiO2 thin film areas are very definite and sharp. Capacitance - Voltage measurements made on TiO2 films gave a dielectric constant of 29, suggesting a possibility of electronic material applications.

• 한국습지학회지
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• 제17권2호
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• pp.139-145
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• 2015

본 연구는 국내에 자생하는 두릅나무과(Araliaceae) 식물 8속 12종을 대상으로 종자의 외부형태적 특성을 조사하고 측정된 형질을 이용하여 다변량분석을 통해 종들간의 유연관계를 밝히기 위해 수행하였다. 종자의 형태적 특성 조사는 각 종의 종자를 20립씩 무작위 선별하여 광학현미경으로 관찰하고 이미지분석시스템을 사용하여 크기를 측정하였다. 측정형질은 종자 크기, 표면 무늬, 광택 유무 등 총 11개 항목이며 이를 이용해 수리분석을 실시하였다. 주성분 분석결과, 전체 변수들 중 제 1주성분과 제 2주성분의 누적기여도는 65.47%이었으며, 제1주성분과 제2주성분에 의해 크게 두릅나무속(Aralia), 오갈피나무속(Eleutherococcus), 인삼속(Panax), 그리고 나머지 기타 속 총 4그룹으로 구분되었다. 군집분석 결과 역시 4그룹으로 구분되어 주성분분석과 동일한 결과를 보였다. 가시오갈피나무(Eleutherococcus senticosus)는 섬오갈피나무(E. gracilistylus)와 유연관계가 가까운 것으로 나타났으며 황칠나무속(Dendropanax)은 오갈피나무속(Eleutherococcus)과 근연관계로 나타났다.

• 한국동물위생학회지
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• 제25권3호
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• pp.245-257
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• 2002

The gene encoding the HN protein from the CBP-1 strain, a heat stable Newcastle disease virus (NDV) isolated from diseased pheasants in Korea, was characterized by reverse transcriptase- polymerase chain reaction(RT-PCR) and the nucleotide and amino acid sequences were analyzed following cloning of the HN gene. In all of the NDV strains studied, a 1.75 kb size cDNA fragment for the HN gene was generated by RT-PCR and smaller specific band sizes harboring the internal portions of the HN gene were also detected by using four pairs of primers. The RT-PCR was sensitive enough to detect viral transcripts when the virus titer was above 25 hemagglutination units. The amplified 1.75 kb cDNA was cloned into a BamHI site of the pVL1393 Baculo transfer vector. The nucleotide sequences of the 1,758 bp HN gene from the CBP-1 strain were determined by the dye terminator cyclic sequencing method. The gene sequences were compared among the strains of CBP-1, Texas GB, Beaudette C, LaSota, B1 and Ulster. The homology of the CBP-1 HN gene to other HN variants was 97.8% to Texas GB, 98.4% to Beaudette C, 95.4% to LaSota, 95.6% to B1 and 90.2% to Ulster. As the deduced 577 amino acid sequences were compared among the strains, the homology for CBP-1 HN appeared to be 96.7% to Texas GB, 97.9% to Beaudette C, 95.5% to LaSota, 95.5% to B1 and 92.7% to Ulster. It was evident that the amino acid sequences included 5 sites for N-asparagine linked glycosylation and 12 cysteine residues. The three conserved leucine residues within the predicted transmembrane domain of the HN protein are amino acid 30, 37 and 44. The three antigenic sites on the HN protein of NDV are amino acids 347(Glu), 481(Asn) and 495(Glu). These data indicate that the genotype of the CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than it is for the LaSota, B1 and Ulster strains.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.14-40
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• 2002

15-Deoxy- Δ$\^$12,14/-prostaglandin J$_2$ (15-deoxy-PGJ$_2$), a naturally occurring ligand activates the peroxisome proliferator-activated receptor-${\gamma}$ (PPAR-${\gamma}$). Activation of PPAR-y has been found to induce cell differentiation such as adipose cell and macrophage. Here it was investigated whether 15-deoxy-PGJ$_2$ has neuronal cell differentiation and possible underlying molecular mechanisms. Dopaminergic differentiating PC 12 cells treated with 15-deoxy-PGJ$_2$ (0.2 to 1.6 ${\mu}$M) alone showed measurable neurite extension and expression of neurofilament, markers of cell differentiation. However much greater extent of neurite extension and expression of neurofilament was observed in the presence of NGF (50 ng/$m\ell$). In parallel with its increasing effect on the neurite extension and expression of neurofilament, 15-deoxy-PGJ$_2$ enhanced NGF-induced p38 MAP kinase expression and its phosphorylation in addition to the activation of transcription factor AP-1 in a dose dependent manner. Moreover, pretreatment of SD 203580, a specific inhibitor of p38 MAP kinase inhibited the promoting effect of 15-deoxy-PGJ$_2$ (0.8 ${\mu}$M) on NGF-induced neurite extension. This inhibition correlated well with the ability of SB203580 to inhibit the enhancing effect of 15-deoxy-PGJ$_2$ on the expression of p38 MAP kinase and activation of AP-1. The promoting ability of 15-deoxy-PGJ$_2$ did not occur through PPAR-${\gamma}$, as synthetic PPAR-${\gamma}$ agonist and antagonist did not change the neurite promoting effect of 15-deoxy-PGJ$_2$. In addition, contrast to other cells (embryonic midbrain and SK-N-MC cells), PPAR-${\gamma}$ was not expressed in PC-12 cells. Other structure related prostaglandins, PGD$_2$ and PGE$_2$ acting via a cell surface G-protein-coupled receptor (GPCR) did not increase basal or NGF-induced neurite extension. Moreover, GPCR (EP and DP receptor) antagonists did not alter the promoting effect of 15-deoxy-PGJ$_2$ on neurite extension and activation of p38 MAP kinase, suggesting that the promoting effect of 15-deoxy-PGJ$_2$ may not be mediated GPCR. These data demonstrate that activation of p38 MAP kinase in conjunction with AP-1 signal pathway may be important in the promoting activity of 15-deoxy-PGJ$_2$ on the differentiation of PC12 cells.

• 생명과학회지
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• 제21권2호
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• pp.235-241
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• 2011

본 연구의 목적은 두유 curd를 형성하는 미생물을 분리하는 것이다. 두유 curd를 형성하는 미생물은 채소를 젖산균으로 발효시킨 전통적인 한국의 음식, 김치로부터 분리하였다. 분리 균주 196개 중 10개의 균주(strain No. 2-2-2, 2-15-2, 2-18-1, 2-19-2, 3-4-1, 3-4-2, 3-8-1, 3-8-3, 3-17-1, 4-39-5)가 단단한 두유 curd를 형성하였고 분자생물학적 생화학적 분석법에 의해 동정되었다. 분리균주로부터 추출한 genomic DNA는 16S rDNA 지역의 PCR 증폭을 위한 주형으로 사용하였다. GenBank 데이터로 16S rDNA 염기서열을 비교한 결과, 분리 균주들은 Leuconostoc mesenteroides group과 Lactobacillus sakei group으로 동정되었다. 두유 curd를 형성하는 균주들의 계통 발생학적 위치와 분류군은 neighbor-joining 방법을 이용하여 확인하였다. 또한, L. mesenteroides group은 생화학적 특성에 의해 L. mesenteroides subsp. dextranicum으로 동정되었다. 하지만 L. sakei group은 생화학적 특성 비교시 다양성을 보여 Lactobacillus sp.로 명명하였다.