• Title/Summary/Keyword: LC2

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

  • Moon, Pyong-Gon;Cho, Young-Eun;Baek, Moon-Chang
    • YAKHAK HOEJI
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    • v.54 no.5
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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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    • v.27 no.1
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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.

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

  • 정영철;김양우;노종수;성낙계;강신권
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.633-639
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    • 1990
  • The genes for cellulases of Pseudomonas sp. LBC505 and CYC10, potent cellulase complex-producing strains, were cloned in Escherichia coli with pUC19. Recombinant plasmids pLCl and pLC2 were isolated from transformants producing cellulase by Congo red staining, and their genes cloned were 0.7 kb and 4.6 kb HindIII fragments, respectively. The inserts of pLCl and pLC2 were hybridized to chromosomal DNAs digested with HindIII from Pseudomona~ sp. LBC505 and CYC10, respectively. Immunodiffusion assays revealed that pLC1-and pLC2-encoded cellulase showed similarity with that of host strains. About 24% of cellulase activity was observed in the extracellular fraction of E. coli carrying pLC1, and its activity was higher about 1.4 times than that of LBC505. The enzymatic properties of pLC1 and pLC2 encoded cellulase were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases were the same as those of cellulase from host strains. HPLC analysis and substrate specificity showed that cellulases cloned were endocellulase.

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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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    • v.12 no.1
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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.

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

  • Kim, Dae-Su;Choi, Jeong-Hwa;Kim, Jong-Han;Park, Soo-Yeon;kang, Seong-In
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.22 no.1
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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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    • v.20 no.4
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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.

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

  • Park, Sun Young;Kim, Ho Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.6
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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.

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

  • Song, Changmin;Jang, Young-Chan
    • Journal of IKEEE
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    • v.24 no.4
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    • pp.975-981
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    • 2020
  • A 10 GHz LC voltage-controlled oscillator (VCO), which controls an amplitude of output signal, is proposed to improve the phase noise. The proposed amplitude control circuit for the LC VCO consists of a peak detector, an amplifier, and a current source. The peak detector is performed detecting the lowest voltage of the output signal by using two diode-connected NMOSFET and a capacitor. The proposed 10 GHz LC VCO with an amplitude control circuit for output signal is designed using a 55 nm CMOS process with a supply voltage of 1.2 V. Its area is 0.0785 ㎟. The amplitude control circuit used in the proposed LC VCO reduces the amplitude variation 242 mV generated in the output signal of the conventional LC VCO to 47 mV. Furthermore, it improves the peak-to-peak time jitter from 8.71 ps to 931 fs.

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

  • Park, Gyo-Beom;Park, Chan Jo;Lee, Sueg-Geun
    • Analytical Science and Technology
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    • v.20 no.5
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    • pp.400-405
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    • 2007
  • The comparative analysis of glycerin in cosmetic samples was carried out by LC/MS and $^1H$ NMR spectrometry. For the LC/MS analysis, aqueous solution was controlled in strong basic condition with sodium hydroxide, and benzoyl chloride was added to the solution for the derivatization of glycerin. The derivative was extracted using pentane and analyzed by the LC/MS. For the $^1H$ NMR analysis, sample was directly dissolved in $D_2O$ solvent without pretreatment. The quantitative analysis of glycerin was done by $^1H$ NMR ERETIC method. The analysis results of LC/MS and $^1H$ NMR showed that the calibration curves were a good linearity with $r^2=0.9991$ in the range of 0.1 to $10{\mu}g/mL$ and $r^2=1$ in the range of 25 to $500{\mu}g/mL$, respectively.

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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    • v.29 no.4
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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).