• Title/Summary/Keyword: time-dependent effect

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Characterization of Insulin-like Growth Factor-free Interaction between Insulin-like Growth Factor Binding Protein 3 and Acid Labile Subunit Expressed from Xenopus Oocytes

  • Choi, Kyung-Yi;Kyung, Yoon-Joo;Lee, Chul-Young;Lee, Dong-Hee
    • BMB Reports
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    • v.37 no.2
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    • pp.153-158
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    • 2004
  • The acid-labile subunit (ALS) is known to interact with the IGF binding protein (IGFBP) in the presence of insulin-like growth factors (IGFs). Studies, however, indicate that ALS forms a doublet with IGFBP3, independent of IGFs. To characterize the structural domain required for the IGF-free ALS-IGFBP3 interaction, seven recombinant human IGFBP3 mutants were generated: three deletion mutants and four site-specific mutants that had altering N-terminal regions of IGFBP3. ALS and IGFBP3 mRNAs were co-injected into Xenopus oocytes, and their products were cross-linked and immunoprecipitated using antisera against ALS or IGFBP3. Among the deletion mutants, the mutant of D40 (deleted in 11-40th amino acids) exerted no effect in the interaction with ALS, while D60 (${\Delta}11$-60) demonstrated a moderate reduction. D88 (${\Delta}11$-88), however, showed a significant decrease. In the case of site-specific mutants, the mutation that alterated the IGF binding site (codons 56 or 80) exerted a significant reduction in the interaction, whereas codons 72 or 87 showed no significant change in the interaction with ALS. The stability of the ALS-IGFBP3 interaction was analyzed according to a time-dependent mode. Consistent with the binding study, mutants on the IGF binding sites (56 or 80) consistently show a weakness in the ALS-IGFBP3 interaction when compared to the mutants that covered the non-IGF binding sites (72 or 87). This study suggests that the N-terminal of IGFBP3, especially the IGF binding site, plays an important role in interacting with ALS as well as in stabilizing the dual complex, independent of IGFs.

Pyruvate Protection against Endothelial Cytotoxicity Induced by Blockade of Glucose Uptake

  • Chung, Se-Jin;Lee, Se-Hee;Lee, Yong-Jin;Park, Hyoung-Sook;Bunger, Rolf;Kang, Young-Hee
    • BMB Reports
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    • v.37 no.2
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    • pp.239-245
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    • 2004
  • We have previously demonstrated that the redox reactant pyruvate prevents apoptosis in the oxidant model of bovine pulmonary artery endothelial cells (BPAEC), and that the anti-apoptotic mechanism of pyruvate is mediated in part via the mitochondrial matrix compartment. However, cytosolic mechanisms for the cytoprotective feature of pyruvate remain to be elucidated. This study investigated the pyruvate protection against endothelial cytotoxicity when the glycolysis inhibitor 2-deoxy-D-glucose (2DG) was applied to BPAEC. Millimolar 2DG blocked the cellular glucose uptake in a concentration- and time-dependent manner with >85% inhibition at $\geq$5 mM within 24 h. The addition of 2DG evoked BPAEC cytotoxicity with a substantial increase in lipid peroxidation and a marked decrease in intracellular total glutathione. Exogenous pyruvate partially prevented the 2DG-induced cell damage with increasing viability of BPAEC by 25-30%, and the total glutathione was also modestly increased. In contrast, 10 mM L-lactate, as a cytosolic reductant, had no effect on the cytotoxicity and lipid peroxidation that are evoked by 2DG. These results suggest that 2DG toxicity may be a consequence of the diminished potential of glutathione antioxidant, which was partially restored by exogenous pyruvate but not L-lactate. Therefore, pyruvate qualifies as a cytoprotective agent for strategies that attenuate the metabolic dysfunction of the endothelium, and cellular glucose oxidation is required for the functioning of the cytosolic glutathione/NADPH redox system.

Effect of Hydrogen Addition on Autoignited Methane Lifted Flames (자발화된 메탄 부상화염에 대한 수소 첨가의 영향)

  • Choi, Byung-Chul;Chung, Suk-Ho
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.36 no.1
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    • pp.75-81
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    • 2012
  • Autoignited lifted flames in laminar jets with hydrogen-enriched methane fuels have been investigated experimentally in heated coflow air. The results showed that the autoignited lifted flame of the methane/hydrogen mixture, which had an initial temperature over 920 K, the threshold temperature for autoignition in methane jets, exhibited features typical of either a tribrachial edge or mild combustion depending on fuel mole fraction and the liftoff height increased with jet velocity. The liftoff height in the hydrogen-assisted autoignition regime was dependent on the square of the adiabatic ignition delay time for the addition of small amounts of hydrogen, as was the case for pure methane jets. When the initial temperature was below 920 K, where the methane fuel did not show autoignition behavior, the flame was autoignited by the addition of hydrogen, which is an ignition improver. The liftoff height demonstrated a unique feature in that it decreased nonlinearly as the jet velocity increased. The differential diffusion of hydrogen is expected to play a crucial role in the decrease in the liftoff height with increasing jet velocity.

Boussinesq Modeling of a Rip Current at Haeundae Beach (Boussinesq 방정식 모형을 이용한 해운대 이안류 수치모의)

  • Choi, Jun-Woo;Park, Won-Kyung;Yoon, Sung-Bum
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.23 no.4
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    • pp.276-284
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    • 2011
  • The rip current occurred at Haeundae beach was numerically investigated under directional random wave environment. The numerical simulation was performed using a fully nonlinear Boussinesq equation model, FUNWAVE which is capable of simulating nearshore circulation since it includes the effect of wave-induced momentum flux and horizontal turbulent mixing. The results of numerical simulation show the time-dependent evolution of the wave-induced nearshore circulation system (including rip current) that are caused by nonlinear transformation of directional irregular waves due to unique topography of Haeundae. From the results, it was found that rip current is well generated and developed where relatively lower wave height and relatively deeper water depth along the longshore direction, and sudden and strong events of rip current were observed.

Responses and Stresses of Structural Vibration of a Camshaft (캠축의 구조 진동 응답 및 응력)

  • Choi, Myung-Jin
    • The Journal of the Acoustical Society of Korea
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    • v.32 no.3
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    • pp.208-213
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    • 2013
  • To get vibration responses, a camshaft is modelled as an unbalanced multiple rotor bearing system. Because of complex geometry and complicated load conditions, the finite element method is used. After the finite element equation of the system is constructed, Newmark's method is used to get the vibration responses. Whirl vibration responses of a V-8 engine camshaft are estimated and compared with measured responses. After the fluctuating stresses are obtained, fatigue analysis is performed based upon the modified Goodman's equation. Stress concentration effects are considered. In the whirl vibration of camshafts, the bending effect is dominant, and the bending deformation is dependent upon the span length between the adjacent bearing journals. For high speeds, the fluctuations of excitation forces are large, and it is known that nonlinear time varying bearing coefficients should be used for analysis.

Imipramine Inhibits A-type Delayed Rectifier and ATP-Sensitive $K^{+}$ Currents Independent of G-Protein and Protein Kinase C in Murine Proximal Colonic Myocytes

  • Choi, Seok;Parajuli, Shankar Prasad;Lim, Geon-Han;Kim, Jin-Ho;Yeum, Cheol-Ho;Yoon, Pyung-Jin;Jun, Jae-Yeoul
    • Archives of Pharmacal Research
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    • v.29 no.11
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    • pp.998-1005
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    • 2006
  • The effects of imipramine on A-type delayed rectifier $K^{+}$ currents and ATP-sensitive $K^{+}\;(K_{ATP)$ currents were studied in isolated murine proximal colonic myocytes using the whole-cell patch-clamp technique. Depolarizing test pulses between-80 mV and +30 mV with 10 mV increments from the holding potential of-80 mV activated voltage-dependent outward $K^{+}$ currents that peaked within 50 ms followed by slow decreasing sustained currents. Early peak currents were inhibited by the application of 4-aminopyridine, whereas sustained currents were inhibited by the application of TEA. The peak amplitude of A-type delayed rectifier $K^{+}$ currents was reduced by external application of imipramine. The half-inactivation potential and the half-recovery time of A-type delayed rectifier $K^{+}$ currents were not changed by imipramine. With 0.1 mM ATP and 140 mM $K^{+}$ in the pipette and 90 mM $K^{+}$ in the bath solution and a holding potential of -80 mV, pinacidil activated inward currents; this effect was blocked by glibenclamide. Imipramine also inhibited $K_{ATP}$ currents. The inhibitory effects of imipramine in A-type delayed rectifier $K^{+}$ currents and $K_{ATP}$ currents were not changed by guanosine 5-O-(2-thiodiphosphate) ($GDP{\beta}S$) and chelerythrine, a protein kinase C inhibitor. These results suggest that imipramine inhibits A-type delayed rectifier $K^{+}$ currents and $K_{ATP}$ currents in a manner independent of G-protein and protein kinase C.

Influence of Quinine on Catecholamine Release Evoked by Cholinergic Stimulation and Membrane Depolarization from the Rat Adrenal Gland

  • Jang, Suk-Jung;Kim, Jong-In;Lim, Dong-Yoon
    • Archives of Pharmacal Research
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    • v.24 no.3
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    • pp.240-248
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    • 2001
  • The present study was attempted to investigate the effect of quinine on secretion of catecholamines (CA) etroked by cholinergic stimulation and membrane depolarization from the isolated perfused rat adrenal gland. The perfusion of quinine (15-150${\mu}$M) into an adrenal vein for 60 min produced dose- and time-dependent inhibition in CA secretion evoked by ACh ($5.32{\times}10^{-3}M$), high $K^{+}5.6{\times}10^{-2}M$, DMPP ($10^{-4}M$ for 2 min), McN-A-343 ($10^{-4}M$ for 2 min), cyclopiazonic acid ($10^{-5}$ for 4 min) and Bay-K-8644 ($10^{-5}$ M for 4 min). Also, under the presence of pinacidil ($10^{-4}$ M), which is also known to be a selective potassium channel activator, CA secretory responses evoked by ACh, high potassium, DMPP McN-A-343, Bay-K-8644 and cyclopiazonic acid were also greatly reduced. When preloaded along with quinine ($5{\times}10^{-5}M$) and glibenclamide ($10^{-6}$ M), a specific blocker of ATP-regulated potassium channels, CA secretory responses evoked by ACh, high potassium, DMPP McN-A-343, Bay-K-8644 and cyclopiazonic acid were recovered as compared to those of quinine-treatment only. taken together, these results demonstrate that quinine inhibits CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by membrane depolarization through inhibiting influx of extracellular calcium and release in intracellular calcium in the rat adrenmodullary chromaffin cells. These findings suggest that activation of potassium channels may be involved at least in inhibitory action of quinine on CA secretion from the rat adrenal gland.

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Effects of Phosphate Complex Treatment on the Quality of Dried Garlic Flakes (인산염 처리가 건마늘 플레이크의 품질에 미치는 영향)

  • Kim, Hyun-Ku;Jo, Kil-Suk;Shin, Dong-Hwa;Kim, Il-Hwan
    • Korean Journal of Food Science and Technology
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    • v.19 no.1
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    • pp.75-80
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    • 1987
  • In order to prevent browning during drying of garlic flakes, the effect of dipping in phosphate complex solution prior to drying on the development of brown pigment in the flakes were studied. The browning color development during drying of the flakes was greatly dependent on phosphate complex concentration and their dipping time. A reciprocal relationship between browning color formation and reducing sugar content of dried garlic flakes was found. The browning color development during drying of the flakes was effectively impeded by dipping in 0.3% phosphate complex solution for 3 minute. The reduction of pyruvate content during drying of the flakes was greatly influenced by phosphate complex concentration and drying temperature, and a maximum retention was acieved by 3 minute dipping in 0.2-0.3% phosphate complex solution. The dried yield of the flakes dipped in phosphate complex solution was 1.62-1.75% lower than control and it was mainly caused by the dissolution of soluble solids during dipping.

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Effects of Citrus sunki Peel Extract on Matrix Metalloproteinase-1 Expression (진귤 과피 추출물의 MMP-1 발현조절 효과)

  • Han, Gu-Seul;Lee, Sun-Ryung
    • Journal of Life Science
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    • v.23 no.12
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    • pp.1553-1556
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    • 2013
  • Flavonoids are one of the major components found in the peels of citrus fruits. Present evidence has suggested that polymethoxyflavonoids, including nobiletin and tangeretin isolated from Citrus sunki, have many biological properties, such as anti-inflammatory, anti-oxidant, and anti-obesity capabilities. Here, we investigated the effect of Citrus sunki peel extract and its possible mechanisms on oxidative stress-induced MMP-1 expression, a major marker of skin photoaging. $H_2O_2$ induced MMP-1 expression in a dose- and time-dependent manner. Extract of Citrus sunki peel (1-25 ${\mu}g/ml$) dose-dependently decreased MMP-1 mRNA levels. When $H_2O_2$ was combined with Citrus sunki peel extract, the phosphorylation of ERK was further decreased compared to a single treatment with $H_2O_2$ alone. Moreover, U0216, an MEK inhibitor, markedly prevented the production of MMP-1. These data suggest that Citrus sunki peel extract has demonstrated protective activity against oxidative damage on MMP-1 expression, and ERK MAP kinase may be involved.

The Role of Kif4A in Doxorubicin-Induced Apoptosis in Breast Cancer Cells

  • Wang, Hui;Lu, Changqing;Li, Qing;Xie, Jun;Chen, Tongbing;Tan, Yan;Wu, Changping;Jiang, Jingting
    • Molecules and Cells
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    • v.37 no.11
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    • pp.812-818
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    • 2014
  • This study was to investigate the mechanism and role of Kif4A in doxorubicin-induced apoptosis in breast cancer. Using two human breast cancer cell lines MCF-7 (with wild-type p53) and MDA-MB-231 (with mutant p53), we quantitated the expression levels of kinesin super-family protein 4A (Kif4A) and poly (ADP-ribose) Polymerase-1 (PARP-1) by Western blot after doxorubicin treatment and examined the apoptosis by flow cytometry after treatment with doxorubicin and PARP-1 inhibitor, 3-Aminobenzamide (3-ABA). Our results showed that doxorubicin treatment could induce the apoptosis of MCF-7 and MDA-MB-231 cells, the down-regulation of Kif4A and upregulation of poly(ADP-ribose) (PAR). The activity of PARP-1 or PARP-1 activation was significantly elevated by doxorubicin treatment in dose- and time-dependent manners (P < 0.05), while doxorubicin treatment only slightly elevated the level of cleaved fragments of PARP-1 (P > 0.05). We further demonstrated that overexpression of Kif4A could reduce the level of PAR and significantly increase apoptosis. The effect of doxorubicin on apoptosis was more profound in MCF-7 cells compared with MDA-MB-231 cells (P < 0.05). Taken together, our results suggest that the novel role of Kif4A in doxorubicin-induced apoptosis in breast cancer cells is achieved by inhibiting the activity of PARP-1.