• Title/Summary/Keyword: SF3B4

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The Uptake of 2-deoxy-D-glucose (2dGlc) by the Endogenous Sugar Transporter(s) of Spodoptera frugiperda Clone 21-AE Cells and the Inhibition of 2dGIc Transport in the Insect Cells by Fructose and Cytoc halasin B

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.177-181
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    • 2003
  • The baculovirus/Spodoptera frugiperda (Sf) cell system has become popular for the production of large amounts of the human erythrocyte glucose transporter, GLUT1, heterologously. However, it was not possible to show that the expressed transporter in insect cells could actually transport glucose. The possible reason for this was that the activity of the endogenous insect glucose transporter was extremely high and so rendered transport activity resulting from the expression of exogenous transporter very difficult to detect. Sf21-AE cells are commonly employed as the host permissive cell line to support the baculovirus AcNPV replication and protein synthesis. The cells grow well on TC-100 medium that contains 0.1 % D-glucose as the major carbon source, strongly suggesting the presence of endogenous glucose transporters. However, unlike the human glucose transporter, very little is known about properties of the endogenous sugar transporter(s) in insect cells. Thus, the uptake of 2-deoxy-D-glucose (2dGlc) by Sf21-AE cells and the inhibition of 2dGlc transport in the insect cells by fructose and cytochalasin B were investigated in the present work. The binding assay of cytochalasin B was also performed, which could be used as a functional assay for the endogenous glucose transporter(s) in the insect cells. Sf21-AE cells were infected with the recombinant virus AcNPV-GT or no virus, at a multiplicity of infection (MOI) of 5. Infected cells were resuspended in PBS plus and minus 300 mM fructose, and plus and minus 20 $\mu$M cytochalasin B for use in transport assays. Uptake was measured at 28$^{\circ}C$ for 1 min, with final concentration of 1 mM deoxy-D-glucose, 2-[1,2-$^3$H]- or glucose, L-[l,$^3$H]-, used at a specific radioactivity of 4 Ci/mol. The results obtained demonstrated that the sugar uptake in uninfected cells was stereospecific, and was strongly inhibited by fructose but only poorly inhibitable by cytochalasin B. It is therefore suggested that the Sf21-AE glucose transporter has very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

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Acupuncture and Herbal Medicine Improved the Quality of life of Obese Women (한방치료를 받은 여성 비만환자의 삶의 질 연구)

  • Jung, Soon-Kwan;Yeom, Seung-Ryong;Kwon, Young-Dal
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.4
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    • pp.1034-1038
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    • 2007
  • To examine the difference of health-related quality of life (QoL) of obese patients between before and after treatment of Korean traditional medicine. This study was assessed in 18 obese-women (age 29.72${\pm}$7.38) treated by acupuncture and herbal medicine using SF-36 quality of life questionnaire between January and July 2006, a 36 item instrument yielding 8 dimensions (Social functioning, Role limitation-Physical, Role limitation-Emotion, Mental health, Vitality, Bodily pain, and General health) and a total score. Their weight was measured by Inbody 3.0 (Biospace co. Korea). QoL and body composition was assessed twice at baseline (B) and 4 weeks after treatment. Auricular acupuncture and electrical needle stimulation to abdomen, thigh, and arm fat was used twice a week and complex herb formula was prescribed according to their constitution and symptoms for 4 weeks. Herb medication was taken 2 times a day. Respective variables were analyzed with Wilcoxon signed ranks test and the level set for statistical significance was p <0.05. The mean of the body weight (P=0.000;B64,68${\pm}$6.86, 4 WKs 60.47${\pm}$5.69), fat percentage (P=0.000;B33.14${\pm}$4.86, 4 WKs 30.16${\pm}$5.34), body mass index (P=0.000;B25.18${\pm}$2.44, 4 WKs 23.46${\pm}$2.09) and fat weight (P=0.000;B21.66${\pm}$5.06, 4 WKs 18.41${\pm}$4.57) of some obese patients decreased significantly between before and after treatment for 4 weeks. Physical functioning (P=0.48;B27.06${\pm}$3.17, 4 WKs 28.00${\pm}$1.71), Mental health (P=0.01 ;B18.83${\pm}$5.25, 4 WKs 22.00${\pm}$3.73), Vitality (P=0.028;B13.89${\pm}$3.03, 4 WKs 15.44${\pm}$2.53), Bodily pain (P=.0014;B8.84${\pm}$1.75, 4 WKs 10.15${\pm}$1.78), and the total scores (P=0.001;B104.99${\pm}$12.60, 4 WKs 114.58${\pm}$11.11) of SF-36 were increased significantly after treatments. These data show some differences in QoL and BMI between before and after treatment on obesity and suggest that the treatment with acupuncture and herbal medicine have a positive impact on several domains of QoL of some obese patients. Further randomized clinical trials (RCTs) including follow-up are needed to examine whether acupuncture and herb medicine have a positive effect on QoL of treatment group compared with control group.

Photoaffinity Labelling of the Human Erythrocyte Glucose Transporters Expressed in Spodoptera frugiperda Clone 9 (Sf9) Cells

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.8 no.4
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    • pp.211-215
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    • 2002
  • The baculovirus/Sf9 cell expression can be employed as a powerful system for producing large amounts of the human erythrocyte glucose transporter, GLUT1 heterologously In order to exploit the system further, it is necessary to develop a convenient method for demonstrating that the transporter expressed in insect cells is biologically active. To achieve this, we have expressed the human CLUT1 in insect cells and photolabelled the expressed protein with [$^3$H] cytochalasin B, a potent inhibitor of the human erythrocyte glucose transporter. Subsequently, the labelled proteins were analysed by SDS-polyacrylamide gel electrophoresis (SDS-PAGE). Membranes labelled with [$^3$H] cytochalasln B in the presence of L-Glucose yielded a single sharp peak of labelling of apparent $M_r$ 45,000 on SDS/polyacrylamide gels. The mobility of this peak corresponded exactly to that of the band detected by anti-glucose transporter antibodies on Western blots of membranes prepared from insect cells infected with recombinant virus. In addition, the sharpness of the radioactive peak provides further evidence for the conclusion that the expressed protein is much less heavily and heterogeneously glycosylated than its erythrocyte counterpart. No peak of labelling was seen with the membranes prepared from non-infected Sf9 cells. Furthermore, the incorporation of label into this peak was completely inhibited by the presence of 500 mM-D-Glucose during tile photolabelling procedure, showing the stereoselectivity of the labelling. These evidences clearly show that human glucose transporter expressed in insect cells exhibits native-like biological activity, and that photolabelling with [$^3$H] cytochalasin B can be a convenient means for analysing the biological activity of the transport protein expressed in insect cells.

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Effect of Truncation of 38 Amino Acids in N-terminal Region of ErmSF, a MLSB Antibiotic Resistance Factor Protein, on Enzymatic Activity (MLSB 항생제 내성인자인 ErmSF의 N-terminal 38개 아미노산 제거가 항생제 내성 효소활성에 미치는 영향)

  • Lee, Hak Jin;Jin, Hyung Jong
    • Korean Journal of Microbiology
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    • v.50 no.3
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    • pp.239-244
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    • 2014
  • ErmSF is one of the four antibiotic resistance factor proteins expressed by Streptomyces fradiae, antibiotic tylosin producer, which renders $MLS_B$ (macrolide-lincosamide-streptogramin B) antibiotic resistance through dimethylating A2058 of 23S rRNA, thereby reducing the affinity of antibiotic to ribosome. Unlike other Erm proteins, ErmSF harbors long N-terminal end region. To investigate its role in enzyme activity, mutant ErmSF deleted of 1-38 amino acids was overexpressed and activity in vivo and in vitro was observed. In vitro enzymatic assay showed that mutant protein exhibited reduced activity by 20% compared to the wild type enzyme. Due to the reduced activity of the mutant protein, cells expressing mutant protein showed weaker resistance to erythromycin than cells with wild type enzyme. Presumably, the decrease in enzyme activity was caused by the hindrance in substrate binding and (or) product release, not by defect in the methyl group transfer occurred in active site.

Clinical Outcomes and Prognosis of Patients with Stent Fracture after Successful Drug-Eluting Stent Implantation (관상동맥 약물 방출 스텐트 삽입 후 스텐트 골절에 대한 임상결과 및 예후)

  • Kim, In-Soo;Han, Jae-Bok;Jang, Seong-Joo
    • Journal of radiological science and technology
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    • v.37 no.2
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    • pp.109-116
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    • 2014
  • Many studies have suggested that in the era of Drug-Eluting Stents(DES) are one of the causes of In-Stent Restenosis(ISR) of Stent Fracture(SF). The present study sought to evaluate clinical characteristics of patients with stent fracture after successful DES implantation. The 4,701 patients were selected for analysis who underwent a follow-up coronary angiography irrespective of ischemic symptoms. The overall incidence of SF was 32 patients(male:female=19:13, Av. age $62.44{\pm}9.8$year, 0.68%). Fractures of Sirolimus-Eluting Stents(SES), Paclitaxel-Eluting Stents(PES), Biolimus A9-Eluting Stents(BES), Everolimus-Eluting Etents(EES), Endothelial Progenitor Cell Capture Stent(EPC) and Zotarolimus-Eluting Stents(ZES) are accounted for 19(59.4%), 9(28.1%), 2(6.3%), 1(3.1%), 1(3.1%) and 0(0%) respectively. SF developed in the left Anterior Dscending(LAD) artery in 16 patients(50%) and in complex(type B2, C) lesions in 25 patients(69.4%). Ten patients were treated with heterogenous DES, the rest being treated with either homogenous DES(3 patients), plain old balloon angioplasty(3 patients), or conservative medical treatment(17 patients). None of the patients with SF suffered from cardiac death during a follow-up period of $32.9{\pm}12.4$ months. The overall rate of DES fracture over up to 3.7 years of follow-up was 0.68% with higher incidence in SES than in PES. SF frequently occurred in the LAD artery and in complex lesions. Of the patients with SF, coronary intervention was performed only when the binary restenosis lesion was significant. During the follow-up, patients with SF have continued on combination antiplatelet therapy. There is a very low rate of major adverse cardiac events(post-detection of SF), especially cardiac death associated with SF.

Nonsaponin fractions of Korean Red Ginseng extracts prime activation of NLRP3 inflammasome

  • Han, Byung-Cheol;Ahn, Huijeong;Lee, Jiseon;Jeon, Eunsaem;Seo, Sanghoon;Jang, Kyoung Hwa;Lee, Seung-Ho;Kim, Cheon Ho;Lee, Geun-Shik
    • Journal of Ginseng Research
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    • v.41 no.4
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    • pp.513-523
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    • 2017
  • Background: Korean Red Ginseng extracts (RGE) have been suggested as effective immune modulators, and we reported that ginsenosides possess anti-inflammasome properties. However, the properties of nonsaponin components of RGE have not been well studied. Methods: To assess the roles of nonsaponin fractions (NS) in NLRP3 inflammasome activation, we treated murine macrophages with or without first or second inflammasome activation signals with RGE, NS, or saponin fractions (SF). The first signal was nuclear factor kappa-light-chain-enhancer of activated B cells (NF-${\kappa}B$)-mediated transcription of pro-interleukin (IL)-$1{\beta}$ and NLRP3 while the second signal triggered assembly of inflammasome components, leading to IL-$1{\beta}$ maturation. In addition, we examined the role of NS in IL-6 production and IL-$1{\beta}$ maturation in mice. Results: NS induced IL-$1{\beta}$ and NLRP3 transcription via toll-like receptor 4 signaling, whereas SF blocked expression. During the second signal, SF attenuated NLRP3 inflammasome activation while NS did not. Further, NS-injected mice presented increased IL-$1{\beta}$ maturation and IL-6 production. Conclusion: SF and NS of RGE play differential roles in the NLRP3 inflammasome activation. Hence, RGE can be suggested as an NLRP3 inflammasome modulator.

Effect of Relative Levels of Mineral Admixtures on Strength of Concrete with Ternary Cement Blend

  • Mala, Kanchan;Mullick, A.K.;Jain, K.K.;Singh, P.K.
    • International Journal of Concrete Structures and Materials
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    • v.7 no.3
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    • pp.239-249
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    • 2013
  • In the present scenario to fulfill the demands of sustainable construction, concrete made with multi-blended cement system of OPC and different mineral admixtures, is the judicious choice for the construction industry. Silica fume (SF) and fly ash (FA) are the most commonly used mineral admixtures in ternary blend cement systems. Synergy between the contributions of both on the mechanical properties of the concrete is an important factor. This study reports the effect of replacement of OPC by fly ash (20, 30, 40 and 50 % replacement of OPC) and/or silica fume (7 and 10 %) on the mechanical properties of concrete like compressive strength and split tensile strength, with three different w/b ratio of 0.3, 0.4 and 0.45. The results indicate that, as the total replacement level of OPC in concrete using ternary blend of OPC + FA + SF increases, the strength with respect to control mix increases up to certain replacement level and thereafter decreases. If the cement content of control mixes at each w/b ratio is kept constant, then as w/b ratio decreases, higher percentage of OPC can be replaced with FA + SF to get 28 days strength comparable to the control mix. A new method was proposed to find the efficiency factor of SF and FA individually in ternary blend cement system, based on principle of modified Bolomey's equation for predicting compressive strength of concrete using binary blend cement system. Efficiency factor for SF and FA were always higher in ternary blend cement system than their respective binary blend cement system. Split tensile strength of concrete using binary and ternary cement system were higher than OPC for a given compressive strength level.

A Study on Etching of Si3N4 Thin Film and the Exhausted Gas Using C3F6 Gas for LCD Process (LCD 공정용 C3F6 가스를 이용한 Si3N4 박막 식각공정 및 배출가스에 관한 연구)

  • Jeon, S.C.;Kong, D.Y.;Pyo, D.S.;Choi, H.Y.;Cho, C.S.;Kim, B.H.;Lee, J.H.
    • Journal of the Korean Vacuum Society
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    • v.21 no.4
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    • pp.199-204
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    • 2012
  • $SF_6$ gas is widely used for dry etching process of semiconductor and display fabrication process. But $SF_6$ gas is considered for typical greenhouse gas for global warming. So it is necessary to research relating to $SF_6$ alternatives reducing greenhouse effect in semiconductor and display. $C_3F_6$ gas is one of the promising candidates for it. We studied about etch characteristics by performing Reactive Ion Etching process of dry etching and reduced gas element exhausted on etching process using absorbent Zeolite 5A. $Si_3N_4$ thin film was deposited to 500 nm with Plasma Enhanced Chemical Vapor Deposition and we performed Reactive Ion Etching process after patterning through photolithography process. It was observed that the etch rate and the etched surface of $Si_3N_4$ thin film with Scanning Electron Microscope pictures. And we measured and compared the exhausted gas before and after the absorbent using Gas Chromatograph-Mass Spectrophotometry.

고전압 펄스 시스템 '천둥'을 이용한 N2, SF6 및 혼합기체에서의 전기 방전 현상 연구

  • Byeon, Yong-Seong;Song, Gi-Baek;Hong, Yeong-Jun;Han, Yong-Gyu;Eom, Hwan-Seop;Choe, Eun-Ha
    • Proceedings of the Korean Vacuum Society Conference
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    • 2010.08a
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    • pp.102-102
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    • 2010
  • 수 Tera Watt급의 가속기 및 펄스파워 시스템은 다수의 스위치를 사용하고 있으며, 이와 같은 가속기 및 시스템의 성능은 기체방전 스위치의 성능에 직접적으로 관련되어 있다. 일반적으로 이와 같은 기체방전, 액체방전 고출력 스위치는 다목적으로 많은 연구와 개발에 응용되고 있다. 예를 들어 천둥 펄스전자빔 발생장치는 12개의 Marx gap 및 3개의 100 kV 펄스충전 전기트리거 gap을 가지고 있다. 기체 방전 또는 액체 방전 펄스 충전 갭 스위치의 음극에 펄스 고전압이 인가되면 이로 인하여 음극에서 전자빔이 발생한다. 내부에는 전자빔이 양극과 충돌하는 순간 양극표면에 플라스마가 형성된다. 이와 같은 플라스마 sheath는 축 방향 이극관 안에서 양극충전 에서 음극으로 팽창하면서 전파하며, 또한 거의 동시에 음극표면에도 플라스마가 형성되어 음극에서 양극으로도 팽창하여 전파하게 된다. 이와 같은 펄스충전 고출력 갭 스위치 안에서 발생되는 방전 플라스마의 특성에 관한 갭 breakdown 과정에 대한 특성연구를 한다. 고출력스위치의 특성 조건으로는 방전전압, 방전시간, jitter 등이 있다. 본 연구에서는 최대전압 600 KV, 최대전류 88 KA, 펄스 폭 60 ns의 특성을 가지는 고전압펄스 시스템 '천둥'을 이용하여 방전 챔버에 고전압 펄스를 인가하고 N2와 SF6 혼합기체 종류와 압력에 따른 방전 현상을 연구하였다. 전극은 구리텅스텐 합금재질의 표준전극을 사용하였고, 전극 간격은 20 mm로 고정하였다. 방전 챔버 압력을 100 torr에서 4 기압까지 변화시켜가며 실험을 진행하였고, N2에 대한 SF6의 혼합비율을 0%~100%까지 변화시키며 실험을 진행하였다. 방전 챔버에는 C-dot probe와 B-dot probe를 설치하여 전압과 전류를 측정하였고, C-dot probe 와 B-dot probe는 각각 Northstar사의 10000:1 고전압 probe와 rogowiski coil을 이용하여 시준 하였다. 실험결과 방전전압은 압력이 증가함에 따라 증가하다가 2 기압 이상에서는 완만히 증가하는 경향을 보였고, SF6 혼합비율은 0~10%까지 급격히 증가하고, 그 이상의 혼합비율에서는 완만히 증가하였다. 방전개시시간은 혼합기체 압력에 따라 증가하며 1기압 이상에서는 급격히 증가 하였다. SF6 혼합비율에 따라서는 1 기압 조건까지는 큰 차이가 없었으나 2 기압부터는 급격히 증가하였다. 안정성을 나타내는 jitter는 SF6 100%일 때 가장 컸으나 혼합기체의 변화에 따른 큰 차이는 없었다.

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Performance of self-compacting geopolymer concrete with and without GGBFS and steel fiber

  • Al-Rawi, Saad;Taysi, Nildem
    • Advances in concrete construction
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    • v.6 no.4
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    • pp.323-344
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    • 2018
  • The study herein reports the impact of Steel Fiber (SF) and Ground Granulated Blast Furnaces slag (GGBFS) content on the fresh and hardened properties of fly ash (FA) based Self-Compacting Geopolymer Concrete (SCGC). Two series of self-compacting geopolymer concrete (SCGC) were formulated with a constant binder content of $450kg/m^3$ and at an alkaline-to-binder (a/b) ratio of 0.50. Fly ash (FA) was substituted with GGBFS with the replacement levels being 0%, 25%, 50%, 75%, and 100% by weight in each SCGC series. Steel fiber (SF) wasn't employed in the assembly of the initial concrete series whereas, within the second concrete series, an SF combination was achieved by a constant additional level of 1% by volume. Fresh properties of mixtures were through an experiment investigated in terms of slump flow diameter, T50 slump flow time, V-funnel flow time, and L-box height ratio. Moreover, the mechanical performance of the SCGCs was evaluated in terms of compressive strength, splitting tensile strength, and fracture toughness. Furthermore, a statistical analysis was applied in order to judge the importance of the experimental parameters, like GGBFS and SF contents. The experimental results indicated that the incorporation of SF had no vital impact on the fresh characteristics of the SCGC mixtures whereas GGBFS aggravated them. However, the incorporation of GGBFS was considerably improved the mechanical properties of SCGCs. Moreover, the incorporation of SF with the total different quantity of GGBFS replacement has considerably increased the mechanical properties of SCGCs, by close to (65%) for the splitting strength and (200%) for compressive strength.