• Title/Summary/Keyword: TMB

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Synthesis and physicochemical characterization of NixZnx-Fe2O4/MWCNT nanostructures as enzyme mimetics with peroxidase-like catalytic activity

  • Salarizadeh, Navvabeh;Sadri, Minoo;Hosseini, Hassan;Sajedi, Reza. H.
    • Carbon letters
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    • v.24
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    • pp.103-110
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    • 2017
  • Carbon-based magnetic nanostructures in several instances have resulted in improved physicochemical and catalytic properties when compared to multi-wall carbon nanotubes (MWCNTs) and magnetic nanoparticles. In this study, magnetic MWCNTs with a structure of $Ni_xZn_xFe_2O_4/MWCNT$ as peroxidase mimics were fabricated by the one-pot hydrothermal method. The structure, composition and morphology of the nanocomposites were characterized with X-ray diffraction (XRD), Fourier transform infrared spectroscopy and transmission electron microscopy. The magnetic properties were investigated with a vibrating sample magnetometer. The peroxidase-like catalytic activity of the nanocomposites was investigated by colorimetric and electrochemical tests with 3,3',5,5'-tetramethylbenzidine (TMB) and $H_2O_2$ as the substrates. The results show that the synthesis of the nanocomposites was successfully performed. XRD analysis confirmed the crystalline structures of the $Ni_xZn_xFe_2O_4/MWCNT$ nanohybrids and MWCNTs. The main peaks of the $Ni_xZn_xFe_2O_4/MWCNT$s crystals were presented. The $Ni_{0.25}Zn_{0.25}Fe_2O_4/MWCNT$ and $Ni_{0.5}Zn_{0.5}Fe_2O_4/MWCNT$ nanocatalysts showed nearly similar physicochemical properties, but the $Ni_{0.5}Zn_{0.5}Fe_2O_4/MWCNT$ nanocatalyst was more appropriate than the $Ni_{0.25}Zn_{0.25}Fe_2O_4/MWCNT$ nanocatalyst in terms of the magnetic properties and catalytic activity. The optimum peroxidase-like activity of the nanocatalysts was obtained at pH 3.0. The $Ni_{0.5}Zn_{0.5}Fe_2O_4/MWCNT$ nanocatalyst exhibited a good peroxidase-like activity. These magnetic nanocatalysts can be suitable candidates for future enzyme-based applications such as the detection of glucose and $H_2O_2$.

MORPHOLOGICAL ANALYSIS OF DEVELOPMENTAL CHANGES IN SOMA AREA OF DIGASTRIC MOTONEURONS IN THE RAT TRIGEMINAL MOTOR NUCLEI (흰쥐의 두힘살근 운동신경세포의 발달과정에 관한 형태학적 분석)

  • Kim, Jae-Hyun;Park, Mi-Hwa;Paik, Sang-Kyoo;Ma, Su-Kyung;Baek, Sang-Heum;Cha, Du-Won
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.2
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    • pp.137-142
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    • 2005
  • To analyze the developmental changes in soma diameters of digastric motoneurons, wheat-germ agglutinin conjugated horseradish peroxidase (WGA-HRP) was injected into the digastric muscle and visualized the retrogradely HRP-labeled motoneurons through tungstate/tetramethylbenzidine (TMB) and following diaminobenzidine (DAB) reactions. The results obtained from Sprague-Dawley rats at postnatal days 1 (P1), 10 (P10) and 30 (P30) indicated as follows: firstly, soma diameters of digastric motoneurons showed unimodal distribution in all postnatal days examined; secondly, the period of P1 to P10 (period 1) showed about 2 times faster growth rate than that of P10 to P30 (period 2); thirdly, the smallest soma examined in each postnatal day exhibited slower growth rate with that of the largest one (increase ratio in soma diameters from P1 to P30, smallest vs. largest = 1.62 : 1.93); Finally, relative growth rates a day showed again that period 1 had faster growth rate than that of period 2. Consequently, developmental changes in soma diameters of digastric motoneurons resulted in very different growth rates between both periods. This implies that the growth of the soma is almost completing within P10 and thereafter growing slowly. The period 1 and 2 are corresponding to sucking and sucking/masticatory period, respectively. Therefore present study providing morphological changes in soma diameters of digastric motoneurons suggests that both periods and their different growth rates of the motoneurons in each period may closely be related with each other.

Biofiltration of soil Vapor Extraction Off-gas from Gasoline Contaminated Soil Using a Compost (퇴비를 이용한 가솔린 오염토양증기추출 배가스의 바이오필터 처리)

  • 남궁완;박준석
    • Journal of Korea Soil Environment Society
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    • v.4 no.3
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    • pp.25-33
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    • 1999
  • The recent emergence of biofiltration as a cost effective waste-gas control technology has stimulated in European countries and the USA. Biofiltration of soil vapor extraction off-gas from gasoline contaminated site was simulated in lab-scale in this study. A filling material used was a compost. This study was conducted to evaluate biofiltration characteristics of the compost material for gasoline off-gas. TPH elimination capacity at the gas loading rate of about 50g/$\textrm{m}^3$((filling material)/hr was circa 40g/$\textrm{m}^3$((filling material)/hr. Removal rate of xylene was the highest among BTEX. while it was the lowest in case of bezene. The maximum elimination capacity of the compost was about 1.5g/$\textrm{m}^3$((filling material)/hr for benzene. More than 95% of trimethylbenzene and naphtalene were removed below the loading rate of 0.7g/㎥(filling material)/hr. About 80% of total TPH and BTEX were removed by biodegradation.

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Fabrication of a influenza A H1N1 sensor using ZnO nanostructure (산화아연 나노구조를 이용한 H1N1 인플루엔자 A 바이러스 센서 제작)

  • Jang, Yun-Seok;Park, Jung-Il;Nam, Yoon-Kyung;Pak, Jung-Ho
    • Proceedings of the KIEE Conference
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    • 2011.07a
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    • pp.1690-1691
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    • 2011
  • 본 논문에서는 H1N1 인플루엔자 A 바이러스(influenza A H1N1 virus) 검출을 위한 산화아연 나노구조(zinc oxide nano structure) 기반의 전기화학적 면역센서를 제작하고 그 특성을 분석하였다. H1N1 인플루엔자 A 바이러스는 빠른 전파 속도 때문에 정확하고 빠른 검출이 필요하다. 먼저, 2 $mm^2$의 표면적을 갖는 패턴된 금 전극 위에 열수방식(hydrothermal method)으로 성장시킨 산화아연 나노구조가 선택적으로 형성되도록 리프트-오프(lift-off) 방법을 사용하였다. 0.01 M phosphate buffered saline(pH 7.4)에서 2 ${\mu}g$/mL 농도의 1차 항체를 정전기력에 의해 산화아연 나노구조에 고정화한 후, 10 pg/mL ~ 5ng/mL 농도의 H1N1 항원을 적용하여 포획 항체에 결합시키고 HRP(horseradish peroxidase) 효소가 결합된 검출 항체를 항원에 결합시키는 샌드위치 ELISA법을 이용하였다. HRP와 반응하는 TMB(3,3', 5,5'-tetramethylbenzidine)와 과산화수소가 포함된 acetate buffered 용액(pH 5)을 전해질로 사용하고 순환전압전류 측정법(cyclic voltammetry)으로 센서의 특성을 분석하였다. 측정된 순환전압전류그래프(cyclic voltammogram)에서 H1N1 항원 농도 10 pg/mL ~ 5 ng/mL의 응답 전류는 276.47 ${\pm}$ 21.72 nA (평균 ${\pm}$ 표준편차, n=4) ~ 478.89 ${\pm}$ 6.21 nA로 측정되었고, logarithmic하게 증가하는 응답 전류 특성을 보였다.

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Biomimetic sequestration of $CO_2$ and reformation to $CaCO_3$ using bovine carbonic anhydrase immobilized on SBA-15 (생체모방공학을 이용한 bovine carbonic anhydrase를 SBA-15에 고정화하여 이산화탄소분리와 재구성된 $CaCO_3$ 연구)

  • Vinoba, Mari;Kim, Dae-Hoon;Lim, Kyoung-Soo;Jeong, Soon-Kwan;Alagar, Muthukaruppan
    • Proceedings of the KAIS Fall Conference
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    • 2010.11a
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    • pp.495-499
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    • 2010
  • The biocatalytic capture of $CO_2$, and its precipitationas $CaCO_3$, over bovine carbonic anhydrase (BCA) immobilized on a pore-expanded SBA-15 support was investigated. SBA-15 was synthesized using TMB as a pore expander, and the resulting porous silica was characterized by XRD, BET, IR, and FE-SEM analysis. BCA was immobilized on SBA-15 through various approaches, including covalent attachment (BCA-CA), adsorption (BCA-ADS), and cross-linked enzyme aggregation (BCA-CLEA). The immobilization of BCA on SBA-15 was confirmed by the presence of zinc metal in the EDXS analysis. The effects of pH, temperature, storage stability, and reusability on the biocatalytic performance of BCA were characterized by examining para-nitrophenyl acetate (p-NPA) hydrolysis. The $K_{cat}/K_m$ values for p-NPA hydrolysis were 740.05, 660.62, and $680.11M^{-1}s^{-1}$, respectively, where as $K_{cat}/K_m$ for free BCA was $873.76M^{-1}s^{-1}$. The amount of $CaCO_3$ precipitate was measured quantitatively using anion-selective electrode and was found to be 12.41, 11.82, or 11.28 mg $CaCO_3$/mg for BCA-CLEA, BCA-ADS, or BCA-CA, respectively. The present results indicate that the immobilized BCA-CLEA, BCA-ADS, and BCA-CA are green materials, and are tunable, reusable, and promising biocatalysts for $CO_2$ sequestration.

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Effects of Ichungwhan on the Aging Process (이정환(二精丸)이 노화과정에 미치는 영향)

  • Jeong, Ji-Cheon;Hyun, Min-Kyung
    • The Journal of Internal Korean Medicine
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    • v.26 no.2
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    • pp.379-389
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    • 2005
  • Objectives: It is well known that aging and aging-related diseases are linked to the increased level of oxidative stress caused by reactive oxygen species(ROS) and reactive nitrogen species(RNS). Nonprotein-SH decreases during aging, while substances such as ROS, nitric oxide(NO), peroxynitrite($ONOO^-$), myeloperoxidase(MPO), and dityrosine show a significant increase. This study investigated the effect of Ichungwhan on the aging process by examining its effect on the generation of the above-mentioned substances. Methods: Four comparison groups of SD rats were used. They were 6 month-old rats, 24 month-old rats, and 24 month-old rats fed with food containing 0.1% and 0.3% of Ichungwhan extract. The amount of NO, $ONOO^-$, and ROS in the rats' kidneys were examined using a fluorescence microplate reader. The reagents used for this purpose include: dihydrorhodamine 123 (DHR 123), 2',7' -dichlorodihydrofluorescein, diacetate(DCFDA), and 4,5-diaminofluorescein(DAF-2). A spectrophotometer was used to investigate the reactivity of nonprotein-SH and myeioperoxidase(MPO), using reagents such as trichloroacetic acid(TCA) and tetramethylbenzidine(TMB). The amounts of MPO protein and dityrosine were measued by western blot. Results: The observed effects of Ichungwhan on rats were as follows: increase of nonprotein-SH; effective decrease of RNS level by suppression of the generation system of $ONOO^-$ and NO; decrease of ROS level; decrease of the MPO reactivity and the subsequent reduction of amount of MPO protein; retardation of dityrosine formation. It can be hypothesized, therefore, that Ichungwhan affects both the earlier and later phases of the molecular inflammatory process, and retards the aging process. Conclusions: Empirical evidence in this study supports a role for Ichungwhan in generation mechanisms of aging process-linked substances ROS, NO, $ONOO^-$, nonprotein-SH, MPO and dityrosine. Affects contrary to the aging process observed in rats beg further empiricism to investigate potential application of Ichungwhan as a medication for age-related diseases in humans.

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Performance Evaluation of Semi-Active Tuned Mass Damper for Elastic and Inelastic Seismic Response Control (준능동 동조질량감쇠기의 탄성 및 비탄성 지진응답 제어성능 평가)

  • Lee, Sang-Hyun;Chung, Lan;Woo, Sung-Sik;Cho, Seung-Ho
    • Journal of the Earthquake Engineering Society of Korea
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    • v.11 no.2 s.54
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    • pp.47-56
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    • 2007
  • In this study, tile performance of a passive tuned mass damper (TMD) and a semi-active tuned mass damper (STMD) was evaluated in terms of seismic response control of elastic and inelastic structures under seismic loads. First, elastic displacement spectra were obtained for the damped structures with a passive TMD, which was optimally designed using the frequency and damping ratio presented by previous study, and with a STMD proposed in this study. The displacement spectra confirm that STMD provides much better control performance than passive md with less stroke. Also, the robustness or the TMD was evaluated by off-tuning the frequency of the TMD to that of the structure. Finally, numerical analyses were conducted for an inelastic structure of which hysteresis was described by Bouc-Wen model and the results indicated that the performance of the passive TMD of which design parameters were optimized for a elastic structure considerably deteriorated when the hysteretic portion or the structural responses increased, while the STMD showed about 15-40% more response reduction than the TMD.

Studies on Secretion of Catecholamine Evoked by Caffeine from the Isolated Perfused Rat Adrenal Gland

  • Lim, Dong-Yoon;Lee, Jang-Hee;Kim, Won-Shik;Kim, Soo-Bok;Lee, Eun-Hwa;Lee, Byeong-Joo;Ko, Suk-Tai
    • Archives of Pharmacal Research
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    • v.14 no.1
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    • pp.55-67
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    • 1991
  • The influence of caffeine on secretion of catecholamines (CA) was examined in the isolated perfused rat adrenal gland. Caffeine (0.3 mM) perfused into an adrenal vein of the gland produced a marked increase in secretion of CA. This secretory effect of CA evoked by perfusion of caffeine for one minute was considerably prolonged, lasting for more than 90 minutes. The tachyphylaxis to releasing effect of CA induced by caffeine was observed by repeated perfusion of this drug. The caffeine-evoked CA secretion was markedly inhibited by pretreatment with ouabain, trifluoperazine, TMB-8 and perfusion with calcium-free Krebs solution containing 5 mM EGTA, but was not affected by perfusion of calcium-free Krebs solution without other addition. CA secretion evoked by caffeine was not reduced significantly by pretreatment with chlorisondamine but after the first collection of perfusate for 3 min was clearly inhibited. Interestingly, the caffeine-evoked CA secretion was considerably potentiated by pretreatment with atropine or pirenzepine, but after the first collection for 3 min it was markedly decreased. These experimental results suggest that caffeine causes a marked increase in secretion of CA from the isolated perfused rat adrenal gland by an extracellular calcium-independent exocytotic mechanism. The secretory effect of caffeine may be mainly due to mobilization of calcium from an intracellular calcium pool in the rat chromaffin cells and partly due to stimulation of both muscarinic and nicotinic receptors.

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Mechanism of Apoptosis Induced by Diazoxide, a $K^{+}$ Channel Opener, in HepG2 Human Hepatoma Cells

  • Lee, Yong-Soo
    • Archives of Pharmacal Research
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    • v.27 no.3
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    • pp.305-313
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    • 2004
  • The effect of diazoxide, a $K^{+}$channel opener, on apoptotic cell death was investigated in HepG2 human hepatoblastoma cells. Diazoxide induced apoptosis in a dose-dependent manner and this was evaluated by flow cytometric assays of annexin-V binding and hypodiploid nuclei stained with propidium iodide. Diazoxide did not alter intracellular $K^{+}$concentration, and various inhibitors of $K^{+}$channels had no influence on the diazoxide-induced apoptosis; this implies that $K^{+}$channels activated by diazoxide may be absent in the HepG2 cells. However, diazoxide induced a rapid and sustained increase in intracellular $Ca^{2+}$ concentration, and this was completely inhibited by the extracellular $Ca^{2+}$ chelation with EGTA, but not by blockers of intracellular $Ca^{2+}$ release (dantrolene and TMB-8). This result indicated that the diazoxide-induced increase of intracellular $Ca^{2+}$ might be due to the activation of a Ca2+ influx pathway. Diazoxide-induced $Ca^{2+}$ influx was not significantly inhibited by either voltage-operative $Ca^{2+}$ channel blockers (nifedipinen or verapamil), or by inhibitors of $Na^{+}$, $Ca^{2+}$-exchanger (bepridil and benzamil), but it was inhibited by flufenamic acid (FA), a $Ca^{2+}$-permeable nonselective cation channel blocker. A quantitative analysis of apoptosis by flow cytometry revealed that a treatment with either FA or BAPTA, an intracellular $Ca^{2+}$ chelator, significantly inhibited the diazoxide-induced apoptosis. Taken together, these results suggest that the observed diazoxide-induced apoptosis in the HepG2 cells may result from a $Ca^{2+}$ influx through the activation of $Ca^{2+}$-permeable non-selective cation channels. These results are very significant, and they lead us to further suggest that diazoxide may be valuable for the therapeutic intervention of human hepatomas.

Measuring abutment convergence angles using stereovision dental image processing system

  • Yang, Hong-Seok;Park, Ji-Man;Han, Jung-Suk;Lee, Jai-Bong;Kim, Sung-Hun;Yeo, In-Sung
    • The Journal of Advanced Prosthodontics
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    • v.6 no.4
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    • pp.259-265
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    • 2014
  • PURPOSE. The purpose of this study was to develop a dental image processing system using a three-dimensional (3D) camera and stereovision technology. The reliability of the system for measuring axial wall convergence angles was evaluated. MATERIALS AND METHODS. The new system predicted 3D coordinate points from 2D images and calculated distances and angles between points. Two examiners measured axial wall convergence angles for seven artificial abutments using a traditional tracing-based method (TBM) and the stereovision-based method (SVBM). Five wax abutment models of simplified abutment forms were made and axial wall convergence angles of wax models were measured by both methods. The data were statistically analyzed at the level of significance, 0.05. RESULTS. Intraclass correlation coefficients showed excellent intra-examiner and inter-examiner reliabilities for both methods. Bland-Altman plots and paired t-tests showed significant differences between measurements and true values using TBM; differences were not significant with SVBM. CONCLUSION. This study found that the SVBM reflected true angle values more accurately than a TMB and illustrated an example of 3D computer science applied to clinical dentistry.