• 제목/요약/키워드: LC2

검색결과 2,501건 처리시간 0.032초

단백체 분석을 위한 일차원 및 이차원 역상크로마토그래피의 비교 (Comparison of 2-D RP-RP MS/MS with 1-D RP MS/MS for Proteomic Analysis)

  • 문평곤;조영은;백문창
    • 약학회지
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    • 제54권5호
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    • pp.377-386
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    • 2010
  • Single-dimensional (1-D) and two-dimensional (2-D) LC methods were utilized to separate peptides from various sources followed by MS/MS analysis. Two-dimensional ultra-high performance liquid chromatography is a useful tool for proteome analysis, providing a greater peak capacity than 1-D LC. The most popular 2-D LC approach used today for proteomic research combines strong cation exchange and reversed-phase LC. We have evaluated an alternative mode for 2-D LC of peptides using 2-D RP-RP nano UPLC Q-TOF Mass Spectrometry, employing reversed-phase columns in both separation dimensions. As control experiments, we identified 129 proteins in 1-D LC and 322 proteins in 2-D LC from E. coli extract peptides. Furthermore, we applied this method to rat primary hepatocyte and a total of 170 proteins were identified from 1-D LC, and 527 proteins were identified from all 2-D LC system. The in-depth protein profiling established by this 2-D LC MS/MS from rat primary hepatocyte could be a very useful reference for future applications in regards to drug induced liver toxicity.

Transglutaminase 2 Promotes Autophagy by LC3 Induction through p53 Depletion in Cancer Cell

  • Kang, Joon Hee;Lee, Seon-Hyeong;Cheong, Heesun;Lee, Chang Hoon;Kim, Soo-Youl
    • Biomolecules & Therapeutics
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    • 제27권1호
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    • pp.34-40
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    • 2019
  • Transglutaminase 2 (TGase 2) plays a key role in p53 regulation, depleting p53 tumor suppressor through autophagy in renal cell carcinoma. We found that microtubule-associated protein 1A/1B-light chain 3 (LC3), a hallmark of autophagy, were tightly associated with the level of TGase 2 in cancer cells. TGase 2 overexpression increased LC3 levels, and TGase 2 knockdown decreased LC3 levels in cancer cells. Transcript abundance of LC3 was inversely correlated with level of wild type p53. TGase 2 knockdown using siRNA, or TGase 2 inhibition using GK921 significantly reduced autophagy through reduction of LC3 transcription, which was followed by restoration of p53 levels in cancer cells. TGase 2 overexpression promoted the autophagy process by LC3 induction, which was correlated with p53 depletion in cancer cells. Rapamycin-resistant cancer cells also showed higher expression of LC3 compared to the rapamycin-sensitive cancer cells, which was tightly correlated with TGase 2 levels. TGase 2 knockdown or TGase 2 inhibition sensitized rapamycin-resistant cancer cells to drug treatment. In summary, TGase 2 induces drug resistance by potentiating autophagy through LC3 induction via p53 regulation in cancer.

Pseudomonas sp.의 Cellulase 유전자의 대장균에의 클로닝 및 발현 (Molecular Cloning and Expression of Cellulase of Gene of Pseudomonas sp. in Escherichia coli)

  • 정영철;김양우;노종수;성낙계;강신권
    • 한국미생물·생명공학회지
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    • 제18권6호
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    • pp.633-639
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    • 1990
  • Cellulase 복합체와 xylanase를 동시에 분비하는 Pseudomonas sp. LBC 505와 CYC 10의 cellulase 유전자를 pUC19를 사용하여 E.coli에 클로닝시켰다. Congo red 염색시 노란색 환을 형성하는 대장균 형질전환에서 7.0Kb-와 4.6Kb-HindIII 단편을 함유한 재조합 플라스미드 pLC1과 pLC2를 가각 분리하였다. DNA hybridization 실험에서 pLC1 과 pLC2는 Pseudomonas sp. LBC 505와 CYC 10 유래임이 각각 밝혀졌고, Immunoassay 실험에서도 유사성이 인정되었다. pLC1을 함유하고 있는 대장균은 cellulas의 24를 세포외로 분비하였고, 효소활성은 모균주에 비해 1.4배 증가하였다. pLC1과 pLC2의 효소학적 성질도 모균주와 동일하였으며, 기질특이성과 HPLC로 유리당을 분석한 결과, 클로닝된 유전자는 endo type인 것으로 나타났다.

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Electro-optical Properties of Twisted Nematic Liquid Crystal Cell with Silver Nanowire Network Electrodes

  • Jang, Kyeong-Wook;Han, Jeong-Min;Shon, Jin-Geun
    • Journal of Electrical Engineering and Technology
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    • 제12권1호
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    • pp.284-287
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    • 2017
  • This paper introduces liquid crystal (LC) alignment and its electro-optical properties in the LC cells with silver nanowire (AgNW) networks. The AgNW network was used as an electrode of LC cell as a substitute for an indium-tin-oxide (ITO) film. LC alignment characteristics in the LC cell using AgNW networks, which have two different sheet resistances of $60{\Omega}/m^2$ and $80{\Omega}/m^2$, were observed. The LC alignment characteristics including pretilt angle, LC alignment state, and thermal stability are similar irrespective of sheet resistance of AgNW network. However, twisted-nematic (TN)-LC cell normally operated when using AgNW network with sheet resistance of $80{\Omega}/m^2$. Electrooptical properties of TN-LC cell exhibited competitive performance compared to those of TN-LC cell based on conventional ITO electrode, which allow new approaches to replace conventional ITO electrode in display technology.

인동등(忍冬藤)이 인간 유래 악성 흑색종 세포의 유전자 발현에 미치는 영향 (Effects of Lonicerae Caulis (LC) on Gene Expression of Human melanoma cells)

  • 김대수;최정화;김종한;박수연;강성인
    • 한방안이비인후피부과학회지
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    • 제22권1호
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    • pp.11-32
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    • 2009
  • Objective : This study was designed to investigate anti-cancer and whitening activities (LC). So it was investigated the effects of LC on proliferation rates of melanoma genetic profile by LC. Methods : The genetic profile for the effect of LC on human derived melanoma cell, SK-MEL-2, was measured using microarray technique, and the functional analysis on these genes were conducted. Total 441 genes were up-regulated and 830 genes down-regulated in cells treated with LC. Genes induced or suppressed by LC were all mainly concerned with basic signalling pathways, which are involved in cell growth, differentiation and migration. Especially, many genes, which are related in apoptosis and cell cycle arrest were up-regulated by treatment with LC, and genes related in cell cycle were down-regulated. Result : The network of total protein interactions were identified by using cytoscape program, and some key molecules, such as BCL2L1, SIN3A, SMAD2 and c-myc that can be used for elucidation of therapeutical mechanism of medicine in the future. Conclusion : These results suggest possibility of LC as addition drug and whitening cosmetics. In addition, it was also suggested that related mechanisms are involved in BCL2L1, SIN3A, SMAD2 and c-myc related signalling pathways.

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Src Family Kinase Inhibitor PP2 Induces LC3 Conversion in a Manner That is Uncoupled from Autophagy and Increases Apoptosis in Multidrug-Resistant Cells

  • Kim, Yun-Ki;Ahn, Jun-Ho;Lee, Mi-Chael
    • Biomolecules & Therapeutics
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    • 제20권4호
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    • pp.393-398
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    • 2012
  • Recently, we reported that defective autophagy may contribute to the inhibition of the growth in response to PP2 (4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine), a selective SFK inhibitor, in multidrug-resistant v-Ha-ras-transformed NIH 3T3 cells (Ras-NIH 3T3/Mdr). In this study, we demonstrated that PP2 induces LC3 conversion via a mechanism that is uncoupled from autophagy and increases apoptosis in Ras-NIH 3T3/Mdr cells. PP2 preferentially induced autophagy in Ras-NIH 3T3 cells rather than in Ras-NIH 3T3/Mdr cells as determined by LC3-I to LC3-II conversion and GFP-LC3 fluorescence microscopy. Beclin 1 knockdown experiments showed that, regardless of drug resistance, PP2 induces autophagy via a Beclin 1-dependent mechanism. PP2 induced a conformational change in Beclin 1, resulting in the enhancement of the pro-autophagic activity of Beclin 1, in Ras-NIH 3T3 cells. Further, PI3K inhibition induced by wortmannin caused a significant increase in apoptosis in Ras-NIH 3T3 cells, as demonstrated by flow cytometric analysis of Annexin V staining, implying that autophagy inhibition through PI3K increases apoptosis in response to PP2 in Ras-NIH 3T3 cells. However, despite the fact that wortmannin abrogates PP2-induced GFP-LC3 punctae formation, some LC3 conversion remains in Ras-NIH 3T3/Mdr cells, suggesting that LC3 conversion may occur in an autophagy-independent manner. Taken together, these results suggest that PP2 induces LC3 conversion independent of PI3K, concomitant with the uncoupling of LC3 conversion from autophagy, in multidrug-resistant cells.

야관문(夜關門)이 토끼의 혈관과 음경해면체 수축에 미치는 영향 (Effect of Lespedezea Cuneata on the Contraction of Rabbit Common Carotid Artery and Corpus Cavernosum)

  • 박선영;김호현
    • 동의생리병리학회지
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    • 제27권6호
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    • pp.809-817
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    • 2013
  • The aim of this study was to evaluate the mechanism of vasodilation of Lespedezea cuneata(LC) in rabbit common carotid artery and cavernosal smooth muscle. LC relaxed arterial strips precontracted with norepinephrine and cavernosal strips precontracted with phenylephrine. The arterial relaxation effects of LC was endothelium-dependent. $N{\omega}$-nitro-L-arginine(L-NNA), NOS inhibitor, methylene blue(MB), cGMP inhibitor, indomethacin(IM), cyclo-oxygenase inhibitor and tetraethylammonium chloride(TEA), KCa-channel blocker attenuate the relaxation responses of LC in arterial strips. In $Ca^{2+}$-free krebs-ringer solution, pretreatment of LC extract significantly reduced the contraction induced by addition $Ca^{2+}$. L-NNA reduced LC extract-induced relaxation in cavernosal strips, but IM, TEA and MB didn't affect LC extract-induced relaxation. When LC extract was applicated on human umbilical vein endothelial cell, the nitric oxide concentration was increased. We conclude that in rabbit common carotid artery, LC may suppress influx of extra-cellular $Ca^{2+}$ through the release of endothelium derived relaxing factor including nitric oxide, prostacyclin, endothelium derived hyperpolarizing factor. And LC exerts a relaxing effect on corpus cavernosum through activating the NO.

출력 신호의 진폭 제어 회로를 가진 10 GHz LC 전압 제어 발진기 (10 GHz LC Voltage-controlled Oscillator with Amplitude Control Circuit for Output Signal)

  • 송창민;장영찬
    • 전기전자학회논문지
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    • 제24권4호
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    • pp.975-981
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    • 2020
  • 위상 잡음을 개선하기 위한 출력 신호의 진폭을 제어하는 회로를 가진 10 GHz LC 전압 제어 발진기(VCO : voltage-controlled oscillator)가 제안된다. 제안된 LC VCO를 위한 진폭 제어 회로는 피크 검출 회로, 증폭기, 그리고 전류원 회로로 구성된다. 피크 검출 회로는 2 개의 diode-connected NMOSFET과 하나의 커패시터로 구성되어 출력 신호의 최젓값을 감지함으로 수행된다. 제안하는 진폭 제어 회로를 가진 LC VCO는 1.2 V 공급 전압을 사용하는 55 nm CMOS 공정에서 설계된다. 설계된 LC VCO의 면적은 0.0785 ㎟이다. 제안된 LC VCO에 사용된 진폭 제어 회로는 기존 LC VCO의 출력 신호에서 발생되는 242 mV의 진폭 변화를 47 mV로 줄인다. 또한, 출력 신호의 peak-to-peak 시간 지터를 8.71 ps에서 931 fs로 개선한다.

LC/MS와 1H NMR을 이용한 화장품속의 글리세린 비교분석 (Comparative analysis of glycerin in cosmetics by LC/MS and 1H NMR)

  • 박교범;박찬조;이석근
    • 분석과학
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    • 제20권5호
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    • pp.400-405
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    • 2007
  • 액체크로마토그래피/질량분석법(LC/MS) 및 핵자기공명분광분석법 $^1H(NMR)$을 이용하여 화장품에 들어있는 글리세린을 동시 비교분석하였다. 화장품 시료를 물에 용해시키고 수산화나트륨을 첨가하여 시료용액을 강알칼리 상태로 유지시킨 후, 시료용액 중의 글리세린을 benzoyl chloride로 유도체화 반응 시키고 유도체화된 글리세린을 pentane으로 추출하여 LC/MS로 정량분석 하였다. $^1H$ NMR 분석은 시료를 전처리 없이 $D_2O$ 용매에 직접 용해시키고, 글리세린을 ERETIC(Electronic REference To access In vivo Concentrations) 방법을 이용하여 $^1H$ NMR로 직접 정량분석 하였다. LC/MS 및 NMR 분석결과 LC/MS의 검량선은 $0.1-10{\mu}g/mL$ 농도범위에서 $r^2=0.9991$ 이었고 $^1H$ NMR의 검량선은 $25-500{\mu}g/mL$ 농도범위에서 $r^2=1$의 상관계수를 갖는 좋은 직선성을 얻었다.

Effect of limestone calcined clay cement (LC3) on the fire safety of concrete structures

  • Gupta, Sanchit;Singh, Dheerendra;Gupta, Trilok;Chaudhary, Sandeep
    • Computers and Concrete
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    • 제29권4호
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    • pp.263-278
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    • 2022
  • Limestone calcined clay cement (LC3) is a low carbon alternative to conventional cement. Literature shows that using limestone and calcined clay in LC3 increases the thermal degradation of LC3 pastes and can increase the magnitude of fire risk in LC3 concrete structures. Higher thermal degradation of LC3 paste prompts this study toward understanding the fire performance of LC3 concrete and the associated magnitude of fire risk. For fire performance, concrete prepared using ordinary Portland cement (OPC), pozzolanic Portland cement (PPC) and LC3 were exposed to 16 scenarios of different elevated temperatures (400℃, 600℃, 800℃, and 1000℃) for different durations (0.5 h, 1 h, 2 h, and 4 h). After exposure to elevated temperatures, mass loss, residual ultrasonic pulse velocity (rUPV) and residual compressive strength (rCS) were measured as the residual properties of concrete. XRD (X-ray diffraction), TGA (thermogravimetric analysis) and three-factor ANOVA (analysis of variance) are also used to compare the fire performance of LC3 with OPC and PPC. Monte Carlo simulation has been used to assess the magnitude of fire risk in LC3 structures and devise recommendations for the robust application of LC3. Results show that LC3 concrete has weaker fire performance, with average rCS being 11.06% and 1.73% lower than OPC and PPC concrete. Analysis of 106 fire scenarios, in Indian context, shows lower rCS and higher failure probability for LC3 (95.05%, 2.22%) than OPC (98.16%, 0.22%) and PPC (96.48%, 1.14%). For robust application, either LC3 can be restricted to residential and educational structures (failure probability <0.5%), or LC3 can have reserve strength (factor of safety >1.08).