• Title/Summary/Keyword: MC3T3-E1

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Determination of Electron Spin Relaxation Time of the Gadolinium-Chealted MRI Contrast Agents by Using an X-band EPR Technique (EPR을 통한 상자성 자기공명 조영제의 전자스핀 이완시간의 결정)

  • Sung-wook Hong;Yongmin Chang;Moon-jung Hwang;Il-su Rhee;Duk-Sik Kang
    • Investigative Magnetic Resonance Imaging
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    • v.4 no.1
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    • pp.27-33
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    • 2000
  • Purpose: To determine the electronic spin relaxation times, $T_{le}$, of three commercially available Gd-chelated MR contrast agents, Gd-DTPA, Gd-DTPA-BMA and Gd-DOTA, using Electron Paramagnetic Resonance(EPR) technique. Material and Methods: The paramagnetic MR contrast agents, Gd-DTFA(Magnevist) , Gd-DTFA-BMA(OMNISCAN) and Gd-DOTA(Dotarem), were used for this study, The EPR spectra of these contrast agents, which were prepared 2:1 methanol/water solution, were obtained at low temperatures, from $-160^{\circ}C~20^{\circ}C$. The glassy-state EPR spectra for these contrast agents were then fitted by the simulation spectra generated with different zero-field splitting (ZFS) parameters by a computer simulation program 'GEN', which generates the EPR powder spectrum using a given ZFS in $3{\times}3$ tensor. Finally, the spin relaxation times of the contrast agents were then determined from the $T_{2e}$, D, and E values of the best simulation spectra using the McLachlan's theory of average relaxation rate. Results: The electronic transverse spin relaxation times, $T_{2e}'s$, of Gd-DTPA, Gd-DTPA-BMA and Gd-DOTA were 0.113ns, 0.147ns and 1.81ns respectively. The g-values were 1.9737, 1.9735 and 1.9830 and the electronic spin relaxation times, $T_{1e}'s$, were 18.70ns, 33.40ns and $1.66{\mu}s$, respectively. Conclusion: The results of these studies reconfirm that the paramagnetic MR contrast agents with larger ZFS parameters should have shorter $T_{1e}'s$. Among three contrast agents used for this study, Gd-DOTA chelated with cyclic ligand structure shows better electronic property then the others with linear structure. Thus, it is concluded that the exact determination of ZFS parameters is the important factor in evaluating relaxation enhancement effect of the agents and in developing new contrast agents.

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Expression of taurine transporter and taurine uptake in mouse osteoblast cell lines

  • Naomi Ishido;Nakashima, Emi Nakashima;Kang, Yonug-Sook
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.124.2-124.2
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    • 2003
  • Taurine is present in a variety of tissue and exhibits many important physiological functions in the cell. Although it is known that many tissues mediate taurine transport, its functions of taurine transport in bone have not been identified yet. In the present study, we investigated the expression of taurine transporter (TauT) and taurine uptake using mouse stromal ST2 cells and osteoblast-like MC3T3-E1 cells, which is bone related cells. Detection of TauT MRNA expression in these cells were performed by reverse transcription polymerase chain reaction (RT-PCR). (omitted)

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Taxonomic Studies on Spider Mites (Tetranychidae : Acarina) of Korea II. Spider Mites, Parasitic on Wild Plants (한국산 응애과(거미강 : 진드기목) 의 계통 분류학적 연구 II. 야생 식물에 기생하는 응애류)

  • 이원구;이병훈;김영진;강동원
    • Animal Systematics, Evolution and Diversity
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    • v.3 no.2
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    • pp.64-116
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    • 1987
  • The purpose of the present paper is to record taxonomic notes on several Korean spider mites. The specimens on which this study is based were collected by the authors during the period from May, 1986 to June, 1987. The following species are the spider mites concerned in this paper. 1. Panonychus citri (McGregor), 2. Eotetranychus populi Koch, 3. Eotetranychus smithi Pritchard and Baker, 4. Shizotetranychus bambusae Reck, 5. Schizotetranychus celarius (Banks), 6. Shizotetranychus leguminosus Ehara, 7. Oligonychus orthis Rimando, 8. Tetranychus ranzawai Kishida, 9. Tetranychus phaselus Ehara, 10. Tetranychus trancatus Ehara , 11. Tetranychus vienensis Zacher, 12. Tetranychus urticae Koch . Among them, eight species with asterisk are newly recorded from Korea . In addition, empodiums of pretarsus were carefully observed and enzyme patterns by electrophoresis were compared on some species in order induce their systematic inter-relationship at either species or generic level.

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Anticancer effect of methanol extract of Erigeron bonariensis on oral cancer cells (구강암 세포주에서 실망초 추출물의 항암효능)

  • Jang, Boon-Sil;Lim, Sun-A
    • Journal of Korean society of Dental Hygiene
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    • v.20 no.5
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    • pp.763-771
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    • 2020
  • Objectives: Erigeron bonariensis is a type of Erigeron found throughout the tropical and subtropical areas as one of the perennial plants or pioneer plants. It is known to show detoxifying, antipyretic, and anticancer effects for various cancers. However, there are no reports on the anticancer effect of E. bonariensis on oral cancer cells. In the present study, we investigated the effects of the methanol extract of Erigeron bonariensis (MEEB) on the inhibition of cell growth and induction of apoptosis in mucoepidermoid carcinoma (MEC) cell lines, including the MC3 and YD15 oral cancer cells. Methods: MC3 Cells were treated by dimethyl sulfoxide (DMSO) or methanol extracts of 20 various natural products 20 ㎍/mL for 48 hours and cell viability were analyzed as Trypan blue exclusion assay. The effects of MEEB treatment on the cell viability of MC3 and YD15 cells, for 48 h, were analyzed by Trypan blue exclusion assay. The anticancer efficacy and apoptosis of oral cancer cell lines were analyzed by western blot analysis. The statistical significance of differences between groups was analyzed by Student's two-tailed t-test. A value of P<0.05 compared to the vehicle control was considered statistically significant. Results: Among 20 different naturally derived products, MEEB significantly inhibited cell viability and increased cleaved poly [ADP-ribose] polymerase 1 (PARP) protein in the MC3 and YD15 cells in a concentration-dependent manner. Conclusions: These results suggest that MEEB can be used as a natural anticancer drug for the treatment of human oral cancer.

Anti-Proliferative Activities of Solid-State Fermented Medicinal Herbs Using Phellinus baumii against Human Colorectal HCT116 Cell (장수상황버섯 균사체를 이용한 고체 발효한약재의 대장암 세포성장 억제 활성)

  • Sohn, Ho-Yong;Shin, Yong-Kyu;Kim, Jong-Sik
    • Journal of Life Science
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    • v.20 no.8
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    • pp.1268-1275
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    • 2010
  • This study was carried out to investigate the anti-proliferative activity of solid-state fermented medicinal herbs which include Phellinus baumii. Methanol extracts were prepared from 36 different medicinal herbs and their fermented counterparts. These extracts were used to treat human colorectal HCT116 cell, human embryonic kidney cell HEK-293, pre-adipocyte cell 3T3-L1, and pre-osteoblast cell MC3T3-E1 for 24 hr. At a concentration of 100 ${\mu}g/ml$, the extracts of Amomum villosum, Cnidium officinale Makino, Dendrobium moniliforme, Dictamnus dasycarpus, Diospyros kaki Thunb, Eucommia ulmoides Oliv, Ginkgo biloba L, Magnolia denudata Desrousseaux, Orostachys japonicus, Panax notoginseng, Pharbitis nil Choisy, Polygala tenuifolia and Trichosanthes kirilowii (seed) led to a < 50% decrease in cell proliferation, and mycelium of P. baumii showed a 46.3% decrease in cell proliferation. Meanwhile, the extracts of the 25 fermented herbs showed similar anti-proliferative activities compared to those of individual non-fermented herbs. However, the extracts of the fermented Drynaria fortunei Kunze (1), Lycium chinense Mill (2), Fritillaria thunbergii Miquel (3) and Prunus persica showed increased anti-proliferative activity. The $IC_{50}s$ of (1), (2) and (3) were especially decreased to 28, 85 and 80 ${\mu}g/ml$ from 394, 917 and 149 ${\mu}g/ml$, respectively. Furthermore, the cytotoxicity of the extracts of fermented (1), (2) and (3) against HEK-293, 3T3-L1, and MC3T3-E1was negligible up to 200 ${\mu}g/ml$. These results suggest that solid-state fermentation using the mycellium of P. baumiiproduce potential anti-cancer agents or strengthen the bioactivity of medicinal herbs.

Effects of Glycyrrhiza inflata Batal Extracts on Adipocyte and Osteoblast Differentiation (감초추출물의 지방세포와 조골세포에 대한 분화효과)

  • Seo, Cho-Rong;Byun, Jong Seon;An, Jae Jin;Lee, JaeHwan;Hong, Joung-Woo;Jang, Sang Ho;Park, Kye Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.7
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    • pp.1015-1021
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    • 2013
  • Glycyrrhiza inflata Batal, an important species of licorice, is one of the most widely used medicinal plants for over 4000 years. Glycyrrhiza plant species has been well known for its various therapeutic activities such as anti-inflammatory, anti-allergic, and anti-ulcer. The purpose of this study was to determine the effects of Glycyrrhiza inflata Batal ethanol extracts (GBE) on adipocyte and osteoblast differentiation. Mesenchymal C3H10T1/2 cells were treated with sub-cytotoxic doses of GBE, and its effects on adipocyte differentiation were assessed. We found that GBE dose-dependently increased lipid accumulation and also induced the expression of adipocyte markers, such as $PPAR{\gamma}$ and its target genes, aP2, and adiponectin, in C3H10T1/2 cells. Consistently, similar effects of GBE on lipid accumulation were also observed in preadipocyte 3T3-L1 cells that further supports the pro-adipogenic activities of GBE. We also investigated the effects of GBE on osteoblast differentiation of mesenchymal C3H10T1/2 cells. As a results, we found that GBE increased the activity of alkaline phosphatase in a dose-dependent manner and also promoted the expression of osteoblast markers, such as ALP and RUNX2, during osteoblast differentiation of C3H10T1/2 cells. Similar pro-osteogenic effects of GBE were also observed in preosteoblast MC3T3-E1 cells. Finally, our data show that a major bioactive compound found in Glycyrrhiza inflata Batal, licochalcone A (LA) but not glycyrrhizic acid (GA), can mediate the pro-adipogenic and pro-osteogenic effects of GBE. Taken together, this study provides data to show the possibility of GBE and its bioactive component LA as putative strategies for type 2 diabetes and bone diseases.

Expression and Purification of Recombinant Human Bone Morphogenetic Protein-7 (rhBMP-7) in Bacillus subtilis (고초균을 이용한 재조합 인간 골 형성 단백질-7의 발현과 정제)

  • Kim, Chun-Kwang;Oh, Sung-Duk;Rhee, Jong-Il
    • KSBB Journal
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    • v.25 no.3
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    • pp.257-264
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    • 2010
  • Bone morphogenetic protein-7 (BMP-7) is one of important growth factors for skeletal development and bone growth. In this work, BMP-7 was efficiently expressed in recombinant Bacillus subtilis. The mature BMP-7 protein indicated molecular weight of 15.4 kDa by Western blot assay and was secreted into culture medium with 0.35 ng/mL. The extracellular and intracellular rhBMP-7 proteins were purified by using a FPLC system with an ion exchange column and a gel filtration column. The extracellular and intracellular rhBMP-7 proteins had finally a 57.1% purity and a 36.2% purity, respectively. The purified rhBMP-7 proteins showed an intact biological activity which stimulated alkaline phophatase (ALP) activity in MC3T3-E1 cells.

Preparation of Sulfobetaine Chitosan, Silk Blended Films, and Their Properties (설포베타인 키토산의 실크 블렌드 필름의 제조 및 그들의 성질)

  • Koo, Ja-Sung;Cha, Jae-Ryung;Oh, Se-Heang;Gong, Myoung-Seon
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.54-61
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    • 2014
  • Water-soluble sulfobetaine chitosan (SCs) was prepared for a blending film with Bombyx mori silk fibroin (SF) by reacting chitosan with 1,3-propanesultone. A series of SF/SCs blended films were successfully prepared by mixing aqueous solutions of B. mori SF and SCs. The SF/SCs blended films were examined through spectroscopic and thermal analysis to determine the morphological changes of SF in the SCs. The effects of the SF/SCs blend ratios on physical and mechanical properties were investigated to discover the feasibility of using these films as biomedical materials such as artificial skin and wound dressing. X-ray analysis showed good compatibility between the two biopolymers. The in vitro degradation behavior of the SF/SCs blended films was systematically investigated for up to 8 weeks in phosphate buffered saline solution at $37^{\circ}C$ and showed a mass loss of 46.4% after 8 weeks. All films showed no cytotoxicity by MC3T3-E1 assay. After 3 days of culture, the relative cell number on all the SF/SCs films was slightly lower than that of an optimized tissue culture plastic.

The Effect of ${\Delta}^{12}PGJ2$ and $PPAR{\Gamma}$ Agonist on the Proliferation and Differentiation of Osteoblast ((${\Delta}^{12}PGJ_2$ 및 PPAR 감마 길항체가 조골세포의 증식 및 분화에 미치는 효과)

  • Heo, Jeong-Mi;Kim, Kyoung-Wha;Chung, Kyoung-Wook;Lee, Hye-Joon;Rhyu, In-Chul;Ku, Young;Chung, Chong-Pyoung
    • Journal of Periodontal and Implant Science
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    • v.35 no.1
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    • pp.133-152
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    • 2005
  • 1. 목적 Prostaglandin은 치주질환과 관련된 국소적 골 대사에 중요한 역할을 한다. ${\Delta}^{12}PGJ_2$는 생체 내에서 혈장의 존재 하에 형성되는 천연 $PGD_2$ 대사산물이며 peroxisome- proliferator에 의해 활성화되는 감마 수용체 (PPAR ${\Gamma}$)에 대해 높은 친화성을 갖는 리간드로서 핵 수용체군에 속하는 전사조절인자이다. 이 연구의 목적은 골화 과정에서 ${\Delta}^{12}PGJ_2$의 역할을 규명하기 위해, 조골세포주의 증식과 분화에 미치는 영향과 그에 관련된 세포기전을 조사하는 데에 있다. 2. 방법 인간 골육종세포주인 Saos-2 (ATCC.HTB 85)와 쥐의 조골세포주 (MC3T3-E1)를 배양한 후 실험군에 농도가 각각 $10^{-5}$, $10^{-6}$, $10^{-7}$, $10^{-8}$, $10^{-9}$ 몰인 ${\Delta}^{12}PGJ_2$와 ciglitazone (합성 PPAR 감마 길항체)를 첨가하였다. 조골세포에서 PPAR 감마의 발현을 관찰하기 위해 역전사효소-중합효소연쇄반응(RT-PCR)을 특정한 primer를 이용하여 시행하였다. 세포 증식은 1일, 2일, 3 일째에 MIT 분석법으로 측정하였고, 2 일째에 알칼리성 인산효소 (ALPase) 생산을 측정하였다. 위의 결과에서 얻은 적정한 농도에서 다양한 조골세포 분화의 표지자들-제 1 형 교원질, 알칼리성 인산효소, osteopontin 및 bone sialoprotein-에 대한 간이 정량적 역전사효소-중합효소연쇄반응 (semiquantitative RT-PCR)을 실시하였으며 골결절 형성에 대한 효과를 알아보고자 석회화 분석도 시행하였다. 3. 결과 ${\Delta}^{12}PGJ_2$와 ciglitazone 모두 Saos-2 세포주의 증식을 촉진시켰다 .$10^{-8}$ 몰의 ${\Delta}^{12}PGJ_2$$10^{-6}$몰의 ciglitazone을 첨가한 실험군을 대조군과 비교했을 때, 시간에 비례하여 세포 증식률이 증가되었다. 알칼리성 인산효소의 활성화 검사에서도 증식률에서와 유사한 결과를 보여주었다. 간이 정량적 RT-PCR에서는 ${\Delta}^{12}PGJ_2$로 처리한 군의 경우 제 1 형 교원질, 알칼리성 인산효소, osteopontin, 그리고 bone sialoprotein의 상대적 mRNA 수준이 유의하게 높았다. 석회화 분석에서는 MC3T3-E1 세포를 $10^{-6}$ 몰의 ${\Delta}^{12}PGJ_2$로 처리한 군과 $10^{-5}$ 몰의 ciglitazone으로 처리한 군에서 현저한 골결절 형성을 보였다. 이러한 결과들은 ${\Delta}^{12}PGJ_2$가 유용한 골 유도물질이 될 수 있으며 또한 그 작용기전이 PPAR 감마-의존형 경로와 연관되어 있음을 보여준다.

Inhibitory Effect of Conditioned Medium of Silk Fibroin-Treated Osteoblasts in Osteoclast Differentiation (실크피브로인을 처리한 MC3T3-E1 조골세포 조건배양액의 파골세포 분화억제효과)

  • Yeo, Joo-Hong;Park, Kyung-Ho;Ju, Won-Chul;Lee, Jin-Ah;Lee, Kwang-Gill;Woo, Soon-Ok;Han, Sang-Mi;Kweon, Hae-Yong;Kim, Sung-Su;Cho, Yun-Hi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.8
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    • pp.992-997
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    • 2008
  • In this study, we investigated the indirect effect of silk-fibroin on osteoclastic differentiation of RAW264.7 cells. The conditioned medium were collected from MC3T3-E1 osbeoblasts treated with $0.001\;mg/mL{\sim}0.1\;mg/mL$ silk fibroin for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of osteoclastic cytokines in the conditioned medium, the protein expression of osteoprotegerin (OPG) with silk-fibroin was not significantly different. However, the protein expression of interleukin (IL)-$1{\beta}$ was specifically lower in a dose dependent manner. In RAW264.7 cells, the conditioned medium with silk-fibroin inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrate-resistant alkaline phosphatase (TRAP)-positive osteoclasts in a dose dependent manner. Taken together, we demonstrated that the conditioned medium of silk-fibroin treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with inhibiting selective expression of IL-$1{\beta}$.