• Title, Summary, Keyword: cell migration

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Constitutive Expression of Arylsulfatase from Pseudoalteromonas carageenovora in E. coli and Its Application to Preparation of Agarose (E. coli에서 Pseudoalteromonas carageenovora 유래 Arylsulfatase의 구성적 발현과 Agarose 제조에의 응용)

  • Kim, Mi-Jin;Jang, Yhon-Hwa;Sung, Moon-Hee;Kim, Yeon-Hee;Nam, Soo-Wan
    • Microbiology and Biotechnology Letters
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    • v.35 no.1
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    • pp.11-16
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    • 2007
  • The arylsulfatase gene (astA, 984 bp ORF) from Pseudoalteromonas carrageenovora genome was amplified by PCR and subcloned into the pHCE-IA vector, in which the hyper consitutive expression (HCE) promoter from the D-amino acid aminotransferase (D-AAT) gene of Geobacillus toevii was employed. The transformant cell, Escherichia coli BL21 (DE3)/pHCE-AST, on LB agar plate containig 4-methylumbelliferyl sulfate, showed an intense fluorescence at 360 nm, indicating that 4-methylumbelliferone was liberated by desulfatate activity. When BL21 (DE3)/pHCE-AST was grown on LB media containing 0.4% glucose or 0.4% glycerol, the arylsulfatase activity was higher at glycerol rather than at glucose. On 2% glycerol medium, the arylsulfatase activity reached 15.0 unit/ml, which was 2.6-fold higher expression level than that with 1% glycerol. The DNA ladder in agarose prepared from agar by this recombinant enzyme revealed similar resolution and migration patterns with a commercial agarose. This results suggests that arylsulfatase overexpressed in E. coli could be applicable to the economic production of electrophoretic-grade agarose.

Characteristics of (Ca,Sr)-doped LaCrO3 Coating Layer for Ceramic Interconnect of Solid Oxide Fuel Cell (고체산화물 연료전지용 (Ca,Sr)도핑된 LaCrO3계 세라믹 연결재 코팅층의 특성 연구)

  • Lee, Gil-Yong;Peck, Dong-Hyun;Song, Rak-Hyun
    • Journal of the Korean Electrochemical Society
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    • v.8 no.4
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    • pp.162-167
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    • 2005
  • Using Pechini method, we synthesized the $La_{0.6}Ca_{0.41}CrO_3$ (LCC41) and $La_{0.8}Sr_{0.05}Ca_{0.15}CrO_3$ (LSCC) powders for slurry dip coating, and $La_{0.75}Ca_{0.27}CrO_3$ (LCC27) powder for air plasma spray coating. The sintering property of the powders and their coating properties were investigated. The average particle sizes of the LCC41, LSCC, LCC27 were 0.6, 0.9, $1.5{\mu}m$, respectively. The relative density of LCC41 bulk was to be found about 98%. The LSCC coating on anode support prevented Ca migration of the coated LCC41 on the anode some or less, which was confirmed from EDS result. The air plasma spray-coated LCC27 with the dip-coated LCC41 were more dense and showed better electrical conductivity than those of the air plasma spray-coated LCC27 and the dip-coated LSCC and LSCC41. The LCC41 and LCC27 showed good electrical conductivities, but the LSCC had a poor electrical conductivity probably due to low sinterability

Development of a Continuous Electrolytic System for pH-control with Only One Discharge of Electrolytic Solution by Using Non-equilibrium Steady State Transfer of Ions across Ion Exchange Membranes (이온 교환막에서 이온의 비 평형 정상상태 이동을 이용한 단일 전해액의 배출만을 가지는 pH 조절용 연속식 전해 반응기 개발)

  • Kim Kwang-Wook;Lyu Je-Wook;Kim In-Tae;Park Geun-Il;Lee Eil-Hee
    • Proceedings of the Korean Radioactive Waste Society Conference
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    • pp.101-109
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    • 2005
  • In order to produce only a pH-controlled solution without discharging any unused solution, this work has developed a continuous electrolytic system with a pH-adjustment reservoir being placed before an ion exchange membrane-equipped electrolyzer, where as a target solution was fed into the pH-adjustment reservoir, some portion of the solution in the pH-adjustment reservoir was circulated through the cathodic or anodic chamber of the electrolyzer depending on the type of the ion exchange membrane used, and some other portion of the solution in the pH-adjustment reservoir was discharged from the electrolytic system through other counter chamber with its pH being controlled as acid or base. The phenomena of the pH being controlled in the system could be explained by the electro-migration of the ion species in the solution through the ion exchange membrane under a cell potential difference between anode and cathode and its consequently-occurring non-charge equilibriums and electrolytic water- split reactions in the anodic and cathodic chambers.

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Smad4 Mediated TGF-β/BMP Signaling in Tooth Formation Using Smad4 Conditional Knockout Mouse (치아 발생과정에서 Smad4의 역할)

  • Yoon, Chi-Young;Baek, Jin-A;Cho, Eui-Sic;Ko, Seung-O
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.35 no.2
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    • pp.73-81
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    • 2013
  • Purpose: Smad4 is a central mediator for transforming growth factor-${\beta}$/bone morphogenetic protein ($TGF-{\beta}/BMP$) signals, which are involved in regulating cranial neural crest cell formation, migration, proliferation, and fate determination. Accumulated evidences indicate that $TGF-{\beta}/BMP$ signaling plays key roles in the early tooth morphogenesis. However, their roles in the late tooth formation, such as cellular differentiation and matrix formation are not clearly understood. The objective of this study is to understand the roles of Smad4 in vivo during enamel and dentin formation through tissue-specific inactivation of Smad4. Methods: We generated and analyzed mice with dental epithelium-specific inactivation of the Smad4 gene (K14-Cre:$Smad4^{fl/fl}$) and dental mesenchyme-specific inactivation of Smad4 gene (Osr2Ires-Cre:$Smad4^{fl/fl}$). Results: In the tooth germs of K14-Cre:$Smad4^{fl/fl}$, ameloblast differentiation was not detectable in inner enamel epithelial cells, however, dentin-like structure was formed in dental mesenchymal cells. In the tooth germs of Osr2Ires-Cre:$Smad4^{fl/fl}$ mice, ameloblasts were normally differentiated from inner enamel epithelial cells. Interestingly, we found that bone-like structures, with cellular inclusion, were formed in the dentin region of Osr2Ires-Cre:$Smad4^{fl/fl}$ mice. Conclusion: Taken together, our study demonstrates that Smad4 plays a crucial role in regulating ameloblast and odontoblast differentiation, as well as in regulating epithelial-mesenchymal interactions during tooth development.

Fucoxanthin derivatives from Sargassum siliquastrum inhibit matrix metalloproteinases by suppressing NF-κB and MAPKs in human fibrosarcoma cells

  • Nguyen, Van-Tinh;Qian, Zhong-Ji;Lee, Bonggi;Heo, Soo-Jin;Kim, Kil-Nam;Jeon, You-Jin;Park, Won Sun;Choi, Il-Whan;Jang, Chul Ho;Ko, Seok-Chun;Park, Sun-Joo;Kim, Yong-Tae;Kim, GeunHyung;Lee, Dae-Sung;Yim, Mi-Jin;Je, Jae-Young;Jung, Won-Kyo
    • ALGAE
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    • v.29 no.4
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    • pp.355-366
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    • 2014
  • Fucoxanthin is known to be an effective cell proliferation inhibitor with anti-tumor and anti-angiogenic activities. However, there is a lack of data regarding the biological effects of cis isomers of fucoxanthin. To assess the potential therapeutic properties of 9'-cis-(6'R) fucoxanthin (FcA), and 13-cis and 13'-cis-(6'R) fucoxanthin complex (FcB) isolated from Sarggassum siliquastrum, we investigated their inhibitory effects on matrix metalloproteinases (MMPs) in phorbol 12-myristate 13-acetate (PMA)-induced human fibrosarcoma (HT1080) cells. FcA and FcB reduced MMP-2 and MMP-9 protein and mRNA levels, as well as the migration of these cells, in a dose-dependent manner. Additionally, FcA and FcB increased levels of MMPs inhibition factors such as tissue inhibitor of metalloproteinase-1. FcA and FcB significantly inhibited the transcriptional activity of nuclear factor ${\kappa}B$ (NF-${\kappa}B$) and by inhibiting c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinases. Our results demonstrate that suppression of the NF-${\kappa}B$, JNK, and p38 signaling pathways may inhibit PMA-induced MMP-2 and MMP-9 activity. Therefore, FcA and FcB may be useful in noninvasive therapeutic strategies against fibrosarcoma metastasis.

Development of Scar Improving Materials using Enkephalin Derivatives (엔케팔린 유도체를 이용한 흉터 개선 소재 개발)

  • Kim, Yang Woo;Kim, Hyoung Shik;Kim, Soo-Yun;Choi, Yun-Hee;Moh, Sang Hyun;Cheon, Young Woo
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.8
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    • pp.5336-5342
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    • 2015
  • Although demand for scar treatment has been rising as our quality of life is improved, most scar treatment products rely on importation. Enkephalin is one of the neuropeptides secreted from neuronal ends. As both skin and neuron are derived from the exoderm during the development process, skin cells express opioid receptors as neuronal cells do. Opioid receptors are categorized into three types, mu(m)-, delta(d)-, and kappa(k)- opioid receptors, all of which are directly involved in the wound healing process. In this study, enkephalin derivatives are synthesized by Alanin Scan and their efficacy was evaluated and compared. In vitro wound healing effects, stimulatory effects of collagen synthesis, and skin hydration effects were also evaluated and confirmed. Among Enkephalin derivatives, AS13 showed highest wound healing effect.

Tumor-Derived Transforming Growth Factor-β is Critical for Tumor Progression and Evasion from Immune Surveillance

  • Li, Zheng;Zhang, Li-Juan;Zhang, Hong-Ru;Tian, Gao-Fei;Tian, Jun;Mao, Xiao-Li;Jia, Zheng-Hu;Meng, Zi-Yu;Zhao, Li-Qing;Yin, Zhi-Nan;Wu, Zhen-Zhou
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.13
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    • pp.5181-5186
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    • 2014
  • Tumors have evolved numerous mechanisms by which they can escape from immune surveillance. One of these is to produce immunosuppressive cytokines. Transforming growth factor-${\beta}$(TGF-${\beta}$) is a pleiotropic cytokine with a crucial function in mediating immune suppression, especially in the tumor microenvironment. TGF-${\beta}$ produced by T cells has been demonstrated as an important factor for suppressing antitumor immune responses, but the role of tumor-derived TGF-${\beta}$ in this process is poorly understood. In this study, we demonstrated that knockdown of tumor-derived TGF-${\beta}$ using shRNA resulted in dramatically reduced tumor size, slowing tumor formation, prolonging survival rate of tumor-bearing mice and inhibiting metastasis. We revealed possible underlying mechanisms as reducing the number of myeloid-derived suppressor cells (MDSC) and $CD4^+Foxp3^+$ Treg cells, and consequently enhanced IFN-${\gamma}$ production by CTLs. Knockdown of tumor-derived TGF-${\beta}$ also significantly reduced the conversion of na$\ddot{i}$ve $CD4^+$ T cells into Treg cells in vitro. Finally, we found that knockdown of TGF-${\beta}$ suppressed cell migration, but did not change the proliferation and apoptosis of tumor cells in vitro. In summary, our study provided evidence that tumor-derived TGF-${\beta}$ is a critical factor for tumor progression and evasion of immune surveillance, and blocking tumor-derived TGF-${\beta}$ may serve as a potential therapeutic approach for cancer.

Effects of enamel matrix derivatives on the proliferation and the release of growth factors of human periodontal ligament cells (법랑기질유도체가 인간 치주인대세포의 증식 및 성장인자 발현에 미치는 영향)

  • Jung, Gyu-Un;Pang, Eun-Kyoung
    • The Journal of Korean Academy of Prosthodontics
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    • v.54 no.3
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    • pp.203-209
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    • 2016
  • Purpose: Stimulating the proliferation and migration of periodontal ligament cells (PDLCs) has become the main goal of periodontal regeneration. To accomplish this goal, regeneration procedures have been developed, but results have not been predictable. Recently, tissue engineering using enamel matrix derivatives (EMDs) and growth factors has been applied to periodontal regeneration; however, the mechanism of EMDs is largely unknown. The aim of this study was to investigate the effects of EMDs on the proliferation and release of growth factors from PDLCs. Materials and methods: Human PDLCs were removed from individually extracted 3rd molars of healthy young adults, and cultured in the media containing EMDs (Emdogain, Biora, Malmo, Sweden) at concentration of 0, 12.5, 25, 50, 100, and $200{\mu}g/mL$ each. Cell proliferation and ALP (alkaline phosphatase) activity were measured. The evaluation of growth factors released by PDLCs was also performed by one-way analysis of variance (ANOVA) and Bonferroni's multiple comparison test. Results: Significantly increased proliferation and ALP activity were observed in PDLCs treated with over $25{\mu}g/mL$ and $50{\mu}g/mL$ EMDs, respectively. Additionally, treatment of PDLCs with $50{\mu}g/mL$ resulted in significantly increased release of vascular endothelial growth factor (VEGF) and transforming growth factor $(TGF)-{\beta}$ after 24 h and 48 h, respectively. Conclusion: EMDs enhance the proliferation and ALP activity of PDLCs, and promote the release of growth factors, including VEGF and $TGF-{\beta}$, from PDLCs. Therefore EMDs could be one of the effective methods for periodontal regeneration.

Supplementation of Indigenous Green Microalga (Parachlorella sp.) to Pre-starter Diet for Broiler Chickens (초기 육계 사료내 토착미세조류(Parachlorella sp.) 첨가에 따른 성장 및 면역반응 변화)

  • An, Su Hyun;Joo, Sang Seok;Lee, Hyo Gun;Kim, Z-Hun;Lee, Chang Soo;Kim, Myunghoo;Kong, Changsu
    • Korean Journal of Poultry Science
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    • v.47 no.1
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    • pp.49-59
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    • 2020
  • The present study determined the effect of dietary cultivated microalgae (Parachlorella sp.) on the growth and immune responses of pre-starter broilers. A total of 320 one-day-old birds (Ross 308) were allocated to 4 treatments with 8 blocks in a randomized complete block design. The four experimental diets consisted of a corn-soybean meal-based control diet, and three diets contained 0.5%, 1.0%, and 1.5% microalgae powder at the expense of cornstarch in the control diet. After feeding the experimental diets for 7 days, the body weight and feed intake of all birds were measured, and 8 birds were randomly selected from each treatment. Peripheral blood mononuclear cells (PBMCs) and serum were harvested for immune profile assessment, including cytokines and cell migration receptors. No differences in growth performance were observed among the treatments. The birds that were fed diets containing graded levels of microalga showed a linear increase in the mRNA expression of cytokine genes in PBMCs, including that of IL2, IL1β, and IL18 (P<0.05). With respect to the chemokine receptor genes in PBMCs, mRNA expression of CCR2, CCR9, and ITGA4 changed quadratically (P<0.05), but that of CCR7 increased linearly (P<0.01). Cytokine protein secretion in blood, including that of IL-1β and IL-6, increased linearly (P<0.01) with an increase in the microalgal content. Overall, the present results show that the indigenous microalgae powder used in this study could stimulate immunity with no detrimental effects on the growth performance of pre-starter broiler chickens.

Nonstructural NS5A Protein Regulates LIM and SH3 Domain Protein 1 to Promote Hepatitis C Virus Propagation

  • Choi, Jae-Woong;Kim, Jong-Wook;Nguyen, Lap P.;Nguyen, Huu C.;Park, Eun-Mee;Choi, Dong Hwa;Han, Kang Min;Kang, Sang Min;Tark, Dongseob;Lim, Yun-Sook;Hwang, Soon B.
    • Molecules and Cells
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    • v.43 no.5
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    • pp.469-478
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    • 2020
  • Hepatitis C virus (HCV) propagation is highly dependent on cellular proteins. To identify the host factors involved in HCV propagation, we previously performed protein microarray assays and identified the LIM and SH3 domain protein 1 (LASP-1) as an HCV NS5A-interacting partner. LASP-1 plays an important role in the regulation of cell proliferation, migration, and protein-protein interactions. Alteration of LASP-1 expression has been implicated in hepatocellular carcinoma. However, the functional involvement of LASP-1 in HCV propagation and HCV-induced pathogenesis has not been elucidated. Here, we first verified the protein interaction of NS5A and LASP-1 by both in vitro pulldown and coimmunoprecipitation assays. We further showed that NS5A and LASP-1 were colocalized in the cytoplasm of HCV infected cells. NS5A interacted with LASP-1 through the proline motif in domain I of NS5A and the tryptophan residue in the SH3 domain of LASP-1. Knockdown of LASP1 increased HCV replication in both HCV-infected cells and HCV subgenomic replicon cells. LASP-1 negatively regulated viral propagation and thereby overexpression of LASP-1 decreased HCV replication. Moreover, HCV propagation was decreased by wild-type LASP-1 but not by an NS5A binding-defective mutant of LASP-1. We further demonstrated that LASP-1 was involved in the replication stage of the HCV life cycle. Importantly, LASP-1 expression levels were increased in persistently infected cells with HCV. These data suggest that HCV modulates LASP-1 via NS5A in order to regulate virion levels and maintain a persistent infection.