• Title/Summary/Keyword: FITC

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Inhibitory effects of Nardostachys Jatamansi on the maturation of dendritic cells (감송향이 수지상세포 성숙에 미치는 영향)

  • O, Kwang-Woo;Jeong, Ji-Hye;Cheong, Hyun-Cheol;Cho, Han-Baek;Kim, Song-Baeg;Choe, Chang-Min
    • The Journal of Korean Obstetrics and Gynecology
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    • v.23 no.3
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    • pp.14-25
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    • 2010
  • Purpose: The purpose of this study is to investigate inhibitory effect on the maturation of dendritic cells from aqueous extract from Nardostachys Jatamansi(NJ). Methods: I examined the phenotypic maturation(class II MHC, CD40, CD86), expression of pro-inflammatory cytokine(TNF-$\alpha$, IL-6, IL-12) and endocytosis of FITC-Dextran in the LPS-induced bone marrow-derived dendritic cells(BMDCs) of mice. Furthermore, the Western-blot analysis reveals the mechanism of inhibitory effect. Results: 1. The NJ extract inhibited the phenotypic maturation of BMDCs in a dose-dependent manner. 2. The NJ extract inhibited the LPS induced cytokine production of BMDCs in a dose-dependent manner. 3. The NJ extract enhanced the endocytosis of Dex-FITC in LPS treated DC. 4. The NJ extract inhibited the activation of JNK and p38 phosphorylation, but not ERK phosphorylation of MAPK family and doesn't inhibit Ik-Ba degradation in LPS-stimulated BMDCs. Conclusion: These results suggest that NJ extract is able to attenuate the inflammation and maturation in BMDCs and may inhibit proliferation of T cells. In conclusion, this experiment suggests that NJ extract may be useful in hypersensitivity disease including autoimmune disease.

Phosphorylation of $Ser^{246}$ Residue in Integrin-linked Kinase 1 by Serum- and Glucocorticoid-induced Kinase 1 is Required to Form a Protein-protein Complex with 14-3-3

  • Chun, Jae-Sun;Kang, Sang-Sun
    • Animal cells and systems
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    • v.9 no.3
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    • pp.161-171
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    • 2005
  • Integrin-linked kinase 1 (ILK1) regulates several protein kinases, including PKB/Akt kinase and glycogen synthase kinase ${\beta}$. ILK1 is also involved distinctively in the cell morphological and structural functions by interacting with the components of the extracellular matrix or integrin. According to the information of serum- and glucocorticoid-induced kinase 1 (SGK1) substrate specificity (R-X-R-X-X(S/T)-${\phi};{\phi}$ indicates a hydrophobic amino acid), two putative phosphorylation sites, $Thr^{181}\;and\;Ser^{246}$, were found in ILK1. We showed that ILK1 fusion protein and two fluorescein-labeled ILK1 peptides, $FITC-^{174}RTRPRNGTLN^{183}$ and $FITC-^{239}CPRLRIFSHP^{248}$, were phosphorylated by SGK1 in vitro. We also identified that 14-3-3 ${\theta}\;{\varepsilon}\;and\;{\xi}$, among several 143-3 isotypes $({\beta},\;{\gamma},\;{\varepsilon},\;{\eta},\;{\sigma},\;{\theta},\;{\tau}\;and\;{\xi})$ formed protein complex with ILK1 in COS-1 cells. Furthermore, the phosphorylation of $Ser^{246}$ by SGK1 induced the binding with 14-3-3. It was also demonstrated that 14-3-3-bound ILK1 has reduced kinase activity. Thus, these data suggest that SGK1 phosphorylates $Thr^{181}\;and\;Ser^{246}$ of ILK1 and the phosphorylation of its $Ser^{246}$ makes ILK1 bind to 14-3-3, resulting in the inhibition of ILK1 kinase activity.

Preparation and Characterization of Multilayer Microcapsules using Biocompatible Polymers (생체적합성 고분자를 사용한 다층 조립 구조 캡슐의 제조와 특성)

  • Jeon, Woohong;Kim, Gwang Yeon;Kim, Gue-Hyun;Ha, Chang-Sik
    • Korean Chemical Engineering Research
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    • v.48 no.2
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    • pp.178-184
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    • 2010
  • The aim of this work is the fabrication of polyelectrolyte microcapsules composed of biocompatible polymers such as chitosan, heparin and alginate, to encapsulate the fluorescein isothiocyanate(FITC)-albumin, and to investigate the protein release behavior therefrom. Polyelectrolyte capsules with 4-layer structures could be prepared with biocompatible materials by oppositely charged adsorption using melamin-foramide as a template. Transmission electron microscope(TEM), scanning electron microscope(SEM) and optical microscope confirmed hollow capsule structures. Protein release before and after encapsulation was monitored with a UV-Vis spectrometer. Microcapsules have different behaviors depending on the kind of polyelectrolyte polymers, chitosan-heparin capsules or chitosan-alginate capsules. In conclusion, the polyelectrolyte multilayer shells can be switched between an open and closed state by means of tuning the pH value.

High-fat Diet Accelerates Intestinal Tumorigenesis Through Disrupting Intestinal Cell Membrane Integrity

  • Park, Mi-Young;Kim, Min Young;Seo, Young Rok;Kim, Jong-Sang;Sung, Mi-Kyung
    • Journal of Cancer Prevention
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    • v.21 no.2
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    • pp.95-103
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    • 2016
  • Background: Excess energy supply induces chronic low-grade inflammation in association with oxidative stress in various tissues including intestinal epithelium. The objective of this study was to investigate the effect of high-fat diet (HFD) on intestinal cell membrane integrity and intestinal tumorigenesis in $Apc^{Min/+}$ mice. Methods: Mice were fed with either normal diet (ND) or HFD for 12 weeks. The number of intestinal tumors were counted and biomarkers of endotoxemia, oxidative stress, and inflammation were determined. Changes in intestinal integrity was measured by fluorescein isothiocyanate (FITC)-dextran penetration and membrane gap junction protein expression. Results: HFD group had significantly higher number of tumors compared to ND group (P < 0.05). Blood total antioxidant capacity was lower in HFD group, while colonic 8-hydroxy-2'-deoxyguanosine level, a marker of oxidative damage, was higher in HFD group compared to that of ND group (P < 0.05). The penetration of FITC-dextran was substantially increased in HFD group (P < 0.05) while the expressions of membrane gap junction proteins including zonula occludens-1, claudin-1, and occludin were lower in HFD group (P < 0.05) compared to those in ND group. Serum concentration of lipopolysaccharide (LPS) receptor (CD14) and colonic toll-like receptor 4 (a LPS receptor) mRNA expression were significantly higher in HFD group than in ND group (P < 0.05), suggesting that significant endotoxemia may occur in HFD group due to the increased membrane permeability. Serum interleukin-6 concentration and myeloperoxidase activity were also higher in HFD group compared to those of ND group (P < 0.05). Conclusions: HFD increases oxidative stress disrupting intestinal gap junction proteins, thereby accelerating membrane permeability endotoxemia, inflammation, and intestinal tumorigenesis.

Effect of Fertilization Promoting Peptide (FPP) on the Acrosome Status of Cryopreserved Human Sperm (Fertilization Promoting Peptide (FPP)가 동결 보존 사람정자의 첨체 상태에 미치는 영향)

  • Park, Se Pill;Shin, Hyun Ah;Kim, Eun Young;Lee, Won Don;Lim, Jin Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.149-154
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    • 2005
  • 연구목적: 정자의 첨체상태는 수정능과 상관관계가 있다. 본 연구는 사람 정자의 동결보존 시 Fertilization promoting peptide (FPP) 처리가 첨체 유지에 효과가 있는지를 알아보고자 실시하였다. 연구재료 및 방법: 사람 정자는 정액검사를 의뢰한 시료를 사용하였으며, 적정농도를 조사하기 위하여 25, 50, 100 nM FPP를 신선정자에 처리한 뒤 시간별로 첨체의 변화를 조사하였다. 또한 적정화된 50 nM FPP를 정자의 동결-융해 시에 처리한 뒤 첨체 변화를 조사하였다. 첨체 변화는 FITC - pisum sativum lectin (PSA) 염색방법을 이용하여 조사하였다. 결 과: FPP 농도 변화와 처리시간에 따른 사람 정자의 첨체 변화를 조사하였던 바, 50 nM FPP 처리군에서 대조군보다 높은 온전한 첨체비율을 얻을 수 있었다. 정자의 동결-융해 시, 동결액과 융해액에 50 nM FPP 첨가가 온전한 첨체를 유지하는 비율을 조사하였던 바, 신선 정자의 결과보다는 유의하게 낮지만 무 처리군보다 유의적으로 높은 온전한 첨체를 얻을 수 있는 것을 알 수 있었다. 또한 동결액에만 또는 융해액에만 50 nM FPP 처리를 하더라도 무 처리군보다 유의하게 높은 온전한 첨체 비율을 획득할 수 있음을 알 수 있었다 (p<0.001). 결 론: 사람 정자의 동결보존 시 50 nM FPP 첨가는 자발적으로 발생하는 첨체반응을 억제하고, 온전한 첨체를 유지할 수 있어 수정능 보유에 기여할 수 있을 것으로 사료된다.

Effect of Defined KSOM Medium on the Development of 1-antitrypsin Transgenic Nuclear Transfer Bovine Embryos

  • M.M.U. Bhuiyan;J.K. Cho;G. Jang;Park, E.S.;S.K. Kang;Lee, B.C.;W.S. Hwang
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.74-74
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    • 2002
  • Production of u 1-antitrypsin ($\alpha$AT) in transgenic cows has a great value in the field of medicine. The present study was conducted to determine the effect of chemically defined KSOM media on in vitro development of bovine transgenic nuclear transfer (NT) embryos. An expression plasmid for human $\alpha$AT was constructed by inserting a bovine beta-casein promoter, a green fluorescent protein (GFP) marker gene, and a human $\alpha$AT target gene into a pcDNA3 plasmid. Cumulus cells as donor nuclei in NT were collected from a Holstein cow and transfected by lipid-mediated method using FuGene6 (Roche Molecular Biochemicals, USA) as reagent. GFP expressed cumulus cells were introduced into recipient oocytes under DIC microscopy equipped with FITC filter set. After electrical fusion and chemical activation, reconstructed embryos were cultured in 1) SOF + 0.8% BSA, 2) KSOM + 0.8% BSA, 3) KSOM + 10% FBS and 4) KSOM +0.01% PVA for 192 h at 39$^{\circ}C$ with 5% $CO_2$, 5% $O_2$ and 90% $N_2$in humidified condition. The development of the embryos was recorded and the GFP expression in blastocyst was determined under FITC filter. The average fusion rate was 73.8% (251/340; n=8). The development rates to 2-4 cells, morula, blastocysts and expression rates in blastocysts varied from 70.3 to 76.5%, 30.2 to 33.8%, 25.4 to 33.8% and 11.8 to 15.6%, respectively. The difference in development and expression rates of embryos among 4 culture groups was not significant (P>0.05). This study indicates that chemically defined KSOM medium is also able to support development of bovine transgenic NT embryos at similar rate of SOF or KSOM supplemented with BSA or serum.

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Increasing Effects of Apoptosis When Co-treated Scutellaria barbata D. Don. with Anti-cancer Drugs (반지련(半枝蓮)과 항암제 병용 투여에 의한 암세포 성장 저해에 관한 연구)

  • Nam, Ju-Young;Sung, Jung-Suk;Jun, Hyun-Ik;Lee, Jeong-Won;Kwon, Su-Kyung;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.22 no.1
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    • pp.125-139
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    • 2009
  • Purpose: This experiment was designed to find out increasing effects of S. barbata. co-treatment with anti-cancer drugs at cancer cell's growth inhibition effect. Methods: Divergent observational study of the S. barbata. co-treatment with Cisplatin treatment on HeLa cell. Cell viability using MTT assay, Cell Culture and Cytotoxicity Studies, Cell Cycle Analysis, Annexin V-FITC/PI assay, Cell morphological assessment, PARP cleavage using Western blotting analysis when HeLa cell were co-treated with Cisplatin and Scutellaria Barbata extracts. Results: When HeLa cell were co-treated with Cisplatin and Scutellaria Barbata extracts, we found out viability of HeLa cell, changing in the distribution of cell cycle, Annexin V-FITC staining, DAPI staining, PARP clavage protein assay by Western-blot. So Scutellaria Barbata extracts have increased apoptosis Conclusion: When co-treated Scutellaria Barbata extracts with anti-cancer drugs, the anti-cancer effects were increased. We still not sure which constituent apoptosis at cancer cells and activates anti-cancer effects suppressing, but we believe that it'll be revealed here after with following experiments.

Inhibitory effects of honokiol on LPS and PMA-induced cellular responses of macrophages and monocytes

  • Lee, Sang-Yeol;Cho, Jae-Youl
    • BMB Reports
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    • v.42 no.9
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    • pp.574-579
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    • 2009
  • The regulatory effects of honokiol on the cellular responses of macrophages and monocytes were evaluated. Specifically, we investigated the effects of honokiol with respect to lipopolysaccharide (LPS)-induced cytotoxicity, LPS- or phorbol-12-myristate-13-acetate (PMA)-mediated morphological changes, and relevant events (FITC-dextran-induced phagocytic uptake). Honokiol blocked the LPS-induced cytotoxicity of RAW264.7 cells in a dose-dependent manner. In addition, honokiol appeared to block the production of cytotoxic cytokines such as interleukin (IL)-$1{\beta}$ and tumor necrosis factor (TNF)-$\alpha$, nitric oxide (NO), and reactive oxygen species (ROS). Moreover, honokiol strongly prevented the morphological changes in RAW 264.7 and U937 cells that were induced by LPS and PMA. The surface levels of marker proteins, which are up-regulated under the morphological changes of RAW264.7 and U937 cells, were also diminished. The data presented here strongly suggest that the honokiol modulates various cellular responses managed by macrophages and monocytes.

The Comparative Studies on the Lectins from Kintoki Bean and Taro Tuber (팥콩 Lectin과 토란 Lectin의 특성 비교)

  • Young-Ju Seo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.23 no.3
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    • pp.515-519
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    • 1994
  • The ComparativThe comperisons of Kintoki bean lectin (KBL) and Taro tuber lectin (TTL) which have been studied in our laboratory are summerized. The recoveries of pure lectins are 0.12% and 0.014%, respectively. They seems to have slight differences in isoelectric points(pH) ; 5.19~5.67 for KBL and 6.41~7.42 for TTL. The minimum concentrations of HA are $2.8\mu\textrm{g}/ml\;and\;21.6\mu\textrm{g}/ml$. The enzymatic modification on HA, growth inhibition, inhibition of nutritional absorption and binding capacities (FITC, $^3H$) of KBL are demonstrated to be much greater than those of TTL.

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Inhibitory effect of FLOS CHRYSANTHEMI on the maturation of dendritic cells (감국의 수지상세포의 성숙 억제 효과)

  • Park, Sung-Joo;Choi, Byung-Min;Song, Ho-Joon
    • The Korea Journal of Herbology
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    • v.22 no.3
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    • pp.127-132
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    • 2007
  • Objectives : FLOS CHRYSANTHEMI (FC) has been reported to possess a variety of pharmacological activities. However, the effect of FC on the dendritic cells has not been determined. Methods : To examine the effect of FC on the immune response, we used several methods such as flow cytometric analyses, enzyme-linked immunosorbent assay. Results : 1. FC inhibited lipopolysacchride (LPS)-induced maturation of bone marrow-derived dendritic cells (BMDC) such as down-regulation of MHC class II and CD40. 2. FC also inhibited uptake of FITC-Dextran in BMDC stimulated with LPS. 3. Furthermore, FC inhibited several kinds of cytokine production such as TNF-a, IL-6 and IL-12 in BMDC. Conclusions : These results suggest that FC plays pivotal role m the development of inflammatory diseases.

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