Association of Extracellular Cleavage of E-Cadherin Mediated by MMP-7 with HGF-Induced in vitro Invasion in Human Stomach Cancer Cells

Lee, K.H.,Choi, E.Y.,Hyun, M.S.,Jang, B.I.,Kim, T.N.,Kim, S.W.,Song, S.K.,Kim, J.H.,Kim, J.-R. S. Karger AG 2007 European surgical research Vol.39 No.4

<P><I>Background:</I> Proteolytic shedding of the ectodomain of a variety of transmembrane proteins, including cell-to-cell adhesion molecules, has been observed in solid cancers. We have investigated whether extracellular cleavage of E-cadherin mediated by matrix metalloproteinase-7 (MMP-7) is involved in hepatocyte growth factor (HGF) induced in vitro invasion in stomach cancer cells. <I>Methods:</I> The effects of HGF on the expression of E-cadherin/β-catenin and MMP-7 at both the protein and mRNA levels were assessed in stomach cancer cells, NUGC-3 and MKN-28, and in cells in which the expression of MMP-7 was downregulated by transfection with a MMP-7 short hairpin RNA plasmid. <I>Results:</I> Treatment with HGF increased the extracellular cleavage of E-cadherin and the release of MMP-7 and reduced the level of E-cadherin in a dose- and time-dependent manner. HGF treatment repressed the phosphorylation of β-catenin in a Triton-soluble fraction, but enhanced this phosphorylation in a Triton-insoluble fraction. The association of E-cadherin with β-catenin was decreased by HGF treatment in the Triton-soluble fraction. In addition, treatment of MMP-7 short hairpin RNA transfected NUGC-3 cells with HGF resulted in no extracellular cleavage of E-cadherin and also decreased the in vitro cell invasion. <I>Conclusions:</I> These results suggest that incubation with HGF mediated the release of MMP-7, resulting in extracellular cleavage of E-cadherin from stomach cancer cells. This might be a key mechanism in HGF-induced in vitro invasion and metastasis.</P><P>Copyright © 2007 S. Karger AG, Basel</P>

Direct analysis of site-specific N-glycopeptides of serological proteins in dried blood spot samples

Choi, N. Y.,Hwang, H.,Ji, E. S.,Park, G. W.,Lee, J. Y.,Lee, H. K.,Kim, J. Y.,Yoo, J. S. Springer Science + Business Media 2017 Analytical and Bioanalytical Chemistry Vol.409 No.21

<P>Dried blood spot (DBS) samples have a number of advantages, especially with respect to ease of collection, transportation, and storage and to reduce biohazard risk. N-glycosylation is a major post-translational modification of proteins in human blood that is related to a variety of biological functions, including metastasis, cell-cell interactions, inflammation, and immunization. Here, we directly analyzed tryptic N-glycopeptides from glycoproteins in DBS samples using liquid chromatography-tandem mass spectrometry (LC-MS/MS) without centrifugation of blood samples, depletion of major proteins, desalting of tryptic peptides, and enrichment of N-glycopeptides. Using this simple method, we identified a total of 41 site-specific N-glycopeptides from 16 glycoproteins in the DBS samples, from immunoglobulin gamma 1 (IgG-1, 10 mg/mL) down to complement component C7 (50 mu g/mL). Of these, 32 N-glycopeptides from 14 glycoproteins were consistently quantified over 180 days stored at room temperature. The major abundant glycoproteins in the DBS samples were IgG-1 and IgG-2, which contain nine asialo-fucosylated complex types of 16 different N-glycopeptide isoforms. Sialo-non-fucosylated complex types were primarily detected in the other glycoproteins such as alpha-1-acid glycoprotein 1, 2, alpha-1-antitypsin, alpha-2-macroglobulin, haptoglobin, hemopexin, Ig alpha 1, 2 chain C region, kininogen-1, prothrombin, and serotransferrin. We first report the characterization of site-specific N-glycoproteins in DBS samples by LC-MS/MS with minimal sample preparation.</P>

Analysis of fucosylation in liver-secreted N-glycoproteins from human hepatocellular carcinoma plasma using liquid chromatography with tandem mass spectrometry

Ji, E. S.,Hwang, H.,Park, G. W.,Lee, J. Y.,Lee, H. K.,Choi, N. Y.,Jeong, H. K.,Kim, K. H.,Kim, J. Y.,Lee, S. Springer Science + Business Media 2016 Analytical and Bioanalytical Chemistry Vol.408 No.27

<P>Fucosylation of N-glycoproteins has been implicated in various diseases, such as hepatocellular carcinoma (HCC). However, few studies have performed site-specific analysis of fucosylation in liver-secreted proteins. In this study, we characterized the fucosylation patterns of liver-secreted proteins in HCC plasma using a workflow to identify site-specific N-glycoproteins, where characteristic B- and/or Y-ion series with and without fucose in collision-induced dissociation were used in tandem mass spectrometry. In total, 71 fucosylated N-glycopeptides from 13 major liver-secreted proteins in human plasma were globally identified by LC-MS/MS. Additionally, 37 fucosylated N-glycopeptides were newly identified from nine liver-secreted proteins, including alpha-1-antichymotrypsin, alpha-1-antitrypsin, alpha-2-HS-glycoprotein, ceruloplasmin, alpha-1-acid glycoprotein 1/2, alpha-2-macroglobulin, serotransferrin, and beta-2-glycoprotein 1. Of the fucosylated N-glycopeptides, bi- and tri-antennary glycoforms were the most common ones identified in liver-secreted proteins from HCC plasma. Therefore, we suggest that this analytical method is effective for characterizing fucosylation in liver-secreted proteins.</P>

Allogeneic hematopoietic cell transplantation for myelodysplastic syndrome: prognostic significance of pre-transplant IPSS score and comorbidity

Lee, J-H,Lee, J-H,Lim, S-N,Kim, D-Y,Kim, S H,Lee, Y-S,Kang, Y-A,Kang, S-I,Jeon, M J,Seol, M,Seo, E-J,Chi, H S,Park, C J,Jang, S,Yun, S-C,Lee, K-H Macmillan Publishers Limited 2010 BONE MARROW TRANSPLANTATION -BASINGSTOKE- Vol.45 No.3

We analyzed the clinical significance of pre-transplant International Prognostic Scoring System (IPSS) score and comorbidity in 68 patients who underwent allogeneic hematopoietic cell transplantation (HCT) for myelodysplastic syndrome (MDS) (n=48) or acute myeloid leukemia evolved from MDS (n=20) between December 1995 and January 2008 in a single institute. During a median follow-up period of 41.0 months (range, 3.2–132.0 months), 27 patients died, and 7 relapsed. The 5-year probabilities of overall survival (OS) and event-free survival (EFS) were 60.0 and 57.4%, respectively, and the 5-year cumulative incidences of non-relapse mortality (CINRM) and relapse were 32.7 and 9.9%, respectively. OS, EFS, and CINRM were significantly different according to pre-transplant IPSS score and presence of pre-transplant comorbidity, which were independent risk factors along with Karnofsky performance score in multivariate analyses. In conclusion, pre-transplant IPSS score and comorbidity may stratify the risk of post transplant outcomes in MDS.

How do extreme wet events affect rice quality in a changing climate?

Lee, M.S.,Kang, B.M.,Lee, J.E.,Choi, W.J.,Ko, J.,Choi, J.E.,An, K.N.,Kwon, O.D.,Park, H.G.,Shin, H.R.,Lee, I.,Kim, J.K.,Kim, H.Y. Elsevier 2013 Agriculture, ecosystems & environment Vol.171 No.-

Despite the growing body of evidence that ongoing warming modifies the cereal quality of several crop species, little is known about how it interacts with other climate change drivers. Here, we examined how elevated [CO<SUB>2</SUB>], warming, and extreme wet events (EWEs) would affect the grain quality of a temperate rice (Oryza sativa L.). In 2009 and 2010, plants were grown at two levels of [CO<SUB>2</SUB>] (c. 390 and 650μll<SUP>-1</SUP>) and three levels of warming (ambient, +1.2<SUP>o</SUP>C and +2.2/2.4<SUP>o</SUP>C) over the seasons in six independent temperature gradient field chambers (three each for ambient and elevated [CO<SUB>2</SUB>]). From 20 days after heading, plants were exposed to EWE with a simulated rainfall of 30mm a day and cloud cover by shading (c. 18% of full sunlight) for 10 consecutive days. There were also control plots, which were non-treated (NT) in terms of EWE. We found that warming and EWE, both independently and in their interaction, lead to a significant reduction in the percentages of sound grains, whereas elevated [CO<SUB>2</SUB>] does not seem to do so. Not only warming and EWE, but also elevated [CO<SUB>2</SUB>] resulted in a significant increase of immature chalky grains (ICGs), which mostly consisted of milky-white chalky grains, degrading the grain quality. In addition, warming interacted positively with elevated [CO<SUB>2</SUB>] on ICGs, whereas the warming sensitivity of IGCs was moderated in EWEs: e.g., on average, the % of ICGs in ambient vs. elevated [CO<SUB>2</SUB>] were 8.5% vs. 9.4%, 10.7% vs. 18.1% and 17.8% vs. 29.8% at ambient (24.7<SUP>o</SUP>C), +1.2<SUP>o</SUP>C and +2.4<SUP>o</SUP>C, respectively, whereas they in NT vs. EWE were 1.2% vs. 16.6%, 7.7% vs. 21.1% and 19.4% vs. 28.2%. Our results tightly confirmed that elevated [CO<SUB>2</SUB>] leads to a significant reduction in rice grain protein content (PC). While warming had little effect on the PC, EWE had a significant positive effect, and the EWE-induced increase in PC was smaller in elevated (+3.3%) than in ambient [CO<SUB>2</SUB>] (+10.9%). Given the rice quality predictor of ICGs and PC, our findings imply that expected climate change and weather extremes have a great potential to degrade the quality of rice, in terms of its milling, cooking, eating, and nutritional quality, as well as market value.

Transplantation of Neural-Induced Mesenchymal Stem Cells for Spinal Cord Injury in Dogs

E. Y. Kim,J. H. Lee,K. J. Kim,K. S. Park,Y. B. Park,Y. N. Ha,M. Kim,K. W. Han,X. X Li,J. Yu,D. S. Lee,J. Y. Park,M. K. Kim 한국동물생명공학회(구 한국동물번식학회) 2011 발생공학 국제심포지엄 및 학술대회 Vol.2011 No.1

Cell transplantation therapy using adult stem cells has recently been identified as a potential treatment for spinal cord injury (SCI). But, recovery after traumatic SCI is very limited. As dogs are physiologically much more similar to human compared with other traditional mammalian models in disease presentation and clinical responses, a number of researches demonstrated canis familiaris is a suitable model for human diseases. This study investigated the effect of transplantation of canine Mesenchymal Stem Cells (cMSC) and neural-induced cMSC (nMSC) to understand how these cells improve neurological function in canine SCI model. The differentiation of cMSC into neural precursor cells was induced in dulbecco’s modified eagle’s medium supplemented with N2-supplement, dibutyryl cyclic adenosine monophosphate, and butylated hydroxyanisole. SCI was induced between T1 and T2 by surgical hemi-section in adult dogs, and then assigned to two groups according to the applied cell types (cMSC vs nMSC). Pelleted cMSC or nMSC were transplanted directly into the injured site after SCI, respectively. Analysis of motor function after transplantation was evaluated by modified Olby score. Magnetic resonance imaging (MRI), histological and immunohistichemical analysis were also performed. Functional recovery in group of cMSC was increasing gradually after transplantation and was higher than nMSC. In MRI, we could not confirm any difference between the cMSC and nMSC experimental groups. Immunohistochemically, beta3-tubuline and nestin were observed in injury site of two experimental groups with the expression level close to non-injured groups. Transplantation of mesenchymal stem cells could promote neuronal reconstruction and repair motor function in SCI. These showed mesenchymal stem cells could be a great candidate as a therapeutic tools in degeneration disease, and dogs could be used to explore human regenerative medicine as a promising animal model. This research was supported by iPET (Grants 110056032CG000), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Electrical properties and radiation detector performance of free-standing bulk n-GaN

Lee, I.-H.,Polyakov, A.Y.,Smirnov, N.B.,Govorkov, A.V.,Kozhukhova, E.A.,Zaletin, V.M.,Gazizov, I.M.,Kolin, N.G.,Pearton, S.J. American Vacuum Society 2012 Journal of vacuum science and technology. material Vol.30 No.2

Detection of mechanical complications related to the potential risk of sudden cardiac death in patients with pulmonary arterial hypertension by computed tomography

Lee, S.E.,Im, J.H.,Sung, J.M.,Cho, I.J.,Shim, C.Y.,Hong, G.R.,Chung, N.,Jung, J.W.,Chang, H.J. Elsevier/North-Holland Biomedical Press 2017 INTERNATIONAL JOURNAL OF CARDIOLOGY Vol.243 No.-

Background: We explored the value of cardiac computed tomography (CT) for the detection and prediction of mechanical complications related to the risk of sudden cardiac death (SCD) in pulmonary arterial hypertension (PAH) patients. Methods: PAH patients (n=60, mean age 47+/-15, 31.7% male) with pulmonary artery (PA) enlargement (≥40mm) by echocardiography were studied with cardiac CT. Complications explored were the presence of left main coronary artery (LM) compression, airway compression, PA dissection and PA thrombosis in relation to diameters of main PA (MPA) which were measured in (1) axial plane (MPA<SUB>Ax</SUB>) and (2) LM oblique view (MPA<SUB>LMobq</SUB>). Results: Mechanical complications were found in 21 patients (35.0%): LM compression in 20 patients; airway compression in 3 patients; and PA thrombosis in 4 patients. Patients with complications had more dilated MPA<SUB>LMobq</SUB> than patients without complication (59.4+/-13.0mm vs. 42.4+/-7.0mm, p<0.001). The area under the receiver operating characteristic curve for MPA<SUB>LMobq</SUB> was 0.889 (95% confidence interval: 0.795 to 0.983, p<0.001) with the highest discriminating sensitivity and specificity being 90.5% and 69.2%, respectively at MPA<SUB>LMobq</SUB> of 45mm. MPA<SUB>Ax</SUB> failed to predict the presence of mechanical complications (p>0.05). Conclusion: MPA<SUB>LMobq</SUB>≥45mm was significantly associated with the presence of mechanical complications of PAH. Evaluation with CT should be considered in PAH patients with dilated MPA.

돼지 대장에서 분리한 E.coli의 중금속 내성에 관한 연구

조창현,정욱진,이영수,최인실,강희옥,박남규,김명화,변미경,이현숙 경상대학교 환경보전연구소 1993 環境保全硏究所報 Vol.1 No.1

돼지의 대장에서 서식하는 466개의 대장균 균주들을 분리하여, 중금속인 Ag, Cd, Cu, Hg, Ni 및Pb에 대한 저항성을 조사 하였다. 거의 대부분의 균주들이 이들 여섯가지 중금속 모두에 강한 저항성을 보였다. 이것은 우리 주위환경의 중금속 오염 정도가 심각하다는 것을 간접적으로 나타내는 것이다. Ag, Pb 및 Hg를 함유하고 있는 고체 배지에서 균주를 성장시켰을때, colony 색깔이 중금속 자체의 광택과 같은 색깔을 나타내는 점으로 보아, 이들 중금속에 대한 저항기작은 유해한 중금속 이온을 세포내 에서 무해한 금속 형태로 전환시켜 세포내에 축적시키는 기작임을 시사하였다. 그 중 가장 높은 저항성을 나타내는 isolate 385를 Ag와 Pb를 함유한 액체배지에서 각각 배양한 뒤 세포내 Ag와 Pb 축적량을 조사한 결과, 건조세포 무게당 0.72g Ag 및 0.23g Pb를 세포내에 축적하고 있었다. 따라서, 이 균주를 유전자 조작 등의 방법으로 개발 한다면 산업 폐수내에 존재하는 이 중금속들의 제거에 효율적으로 사용할 수 있을 것이라 사료된다. We isolated E.coli from porcine intestines and examined the resistances to various heavy-metals, Ag, Cd, Cu, Hg, Ni, and Pb. The 466 isolates were resistant to the heavy-metals. Among them, 72.1% was survived in 1 mM AgNO₃, 9.3% in 80 mM AgNO₃, 95.9% in 0.6 mM Cd(NO₃)₂, 5.6% in 3 mN Cd(NO₃)₂. 95.9% in Cu(NO₃)₂, 48.5% in 0.2 mM HgCl₂, 3.4% in 0.6 mM HgCl₂, 64.4% in 5 mM NiCl₂ and 67.4% in 10 mM Pb(NO₃)₂. The isolate 385 was most resistant to silver and lead ions and the MICs of the ions were 80 mM and 11 mM, respectively. These resistances were inducible by Ag^+ and Pb^2+ ions. When isolate 385 grew in LB-agar plates containing AgNO₃ or Pb(NO₃)₂, the colony colors were changed from light yellow to deep brown. This change to brown color suggests that the resistances of 385 cells to Ag^+ and Pb^2+ ions were due to the reducing mechanism which converted them into the elementary metals(Ag^0, Pb^0) after the uptake of the ions into the cells. The resistant cells accumulated 0.72gr of Ag^0 and 0.23gr of Pb^0 per cells dry wt.

Adipose Natural Killer Cells Regulate Adipose Tissue Macrophages to Promote Insulin Resistance in Obesity

Lee, B.C.,Kim, M.S.,Pae, M.,Yamamoto, Y.,Eberle, D.,Shimada, T.,Kamei, N.,Park, H.S.,Sasorith, S.,Woo, J.,You, J.,Mosher, W.,Brady, Hugh J.M.,Shoelson, Steven E.,Lee, J. Cell Press 2016 Cell metabolism Vol.23 No.4

<P>Obesity-induced inflammation mediated by immune cells in adipose tissue appears to participate in the pathogenesis of insulin resistance. We show that natural killer (NK) cells in adipose tissue play an important role. High-fat diet (HFD) increases NK cell numbers and the production of proinflammatory cytokines, notably TNF alpha, in epididymal, but not subcutaneous, fat depots. When NK cells were depleted either with neutralizing antibodies or genetic ablation in E4bp4(+/-) mice, obesity-induced insulin resistance improved in parallel with decreases in both adipose tissue macrophage (ATM) numbers, and ATM and adipose tissue inflammation. Conversely, expansion of NK cells following IL-15 administration or reconstitution of NK cells into E4bp4(-/-) mice increased both ATM numbers and adipose tissue inflammation and exacerbated HFD-induced insulin resistance. These results indicate that adipose NK cells control ATMs as an upstream regulator potentially by producing proinflammatory mediators, including TNF alpha, and thereby contribute to the development of obesity-induced insulin resistance.</P>