• Title/Summary/Keyword: Cu2+ ions

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Purification and Characterization of Mitochondrial Mg2+-Independent Sphingomyelinase from Rat Brain

  • Jong Min Choi;Yongwei Piao;Kyong Hoon Ahn;Seok Kyun Kim;Jong Hoon Won;Jae Hong Lee;Ji Min Jang;In Chul Shin;Zhicheng Fu;Sung Yun Jung;Eui Man Jeong;Dae Kyong Kim
    • Molecules and Cells
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    • v.46 no.9
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    • pp.545-557
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    • 2023
  • Sphingomyelinase (SMase) catalyzes ceramide production from sphingomyelin. Ceramides are critical in cellular responses such as apoptosis. They enhance mitochondrial outer membrane permeabilization (MOMP) through self-assembly in the mitochondrial outer membrane to form channels that release cytochrome c from intermembrane space (IMS) into the cytosol, triggering caspase-9 activation. However, the SMase involved in MOMP is yet to be identified. Here, we identified a mitochondrial Mg2+-independent SMase (mt-iSMase) from rat brain, which was purified 6,130-fold using a Percoll gradient, pulled down with biotinylated sphingomyelin, and subjected to Mono Q anion exchange. A single peak of mt-iSMase activity was eluted at a molecular mass of approximately 65 kDa using Superose 6 gel filtration. The purified enzyme showed optimal activity at pH of 6.5 and was inhibited by dithiothreitol and Mg2+, Mn2+, Ni2+, Cu2+, Zn2+, Fe2+, and Fe3+ ions. It was also inhibited by GW4869, which is a non-competitive inhibitor of Mg2+-dependent neutral SMase 2 (encoded by SMPD3), that protects against cytochrome c release-mediated cell death. Subfractionation experiments showed that mt-iSMase localizes in the IMS of the mitochondria, implying that mt-iSMase may play a critical role in generating ceramides for MOMP, cytochrome c release, and apoptosis. These data suggest that the purified enzyme in this study is a novel SMase.

Purification and Biochemical Characterization of a Novel Fibrinolytic Enzyme from Streptomyces sp. P3

  • Cheng, Guangyan;He, Liying;Sun, Zhibin;Cui, Zhongli;Du, Yingxiang;Kong, Yi
    • Journal of Microbiology and Biotechnology
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    • v.25 no.9
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    • pp.1449-1459
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    • 2015
  • A novel proteolytic enzyme with fibrinolytic activity, FSP3, was purified from the recently isolated Streptomyces sp. P3, which is a novel bacterial strain isolated from soil. FSP3 was purified to electrophoretic homogeneity by ammonium sulfate precipitation, anion exchange, and gel filtration. FSP3 is considered to be a single peptide chain with a molecular mass of 44 kDa. The maximum activity of the enzyme was observed at 50℃ and pH 6.5, and the enzyme was stable between pH 6 and 8 and below 40℃. In a fibrin plate assay, FSP3 showed more potent fibrinolytic activity than urokinase, which is a clinical thrombolytic agent acting as a plasminogen activitor. The activity was strongly inhibited by the serine protease inhibitor PMSF, indicating that it is a serine protease. Additionally, metal ions showed different effects on the activity. It was significantly suppressed by Mg2+ and Ca2+ and completely inhibited by Cu2+, but slightly enhanced by Fe2+. According to LC-MS/MS results, its partial amino acid sequences are significantly dissimilar from those of previously reported fibrinolytic enzymes. The sequence of a DNA fragment encoding FSP3 contained an open reading frame of 1287 base pairs encoding 428 amino acids. FSP3 is a bifunctional enzyme in nature. It hydrolyzes the fibrin directly and activates plasminogen, which may reduce the occurrence of side effects. These results suggest that FSP3 is a novel serine protease with potential applications in thrombolytic therapy.

d10 Metal Complexes of a Tripodal Amine Ligand

  • Choi, Kyu-Seong;Kang, Dong-hyun;Lee, Ji-Eun;Seo, Joo-beom;Lee, Shim-Sung
    • Bulletin of the Korean Chemical Society
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    • v.27 no.5
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    • pp.747-750
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    • 2006
  • Research on tripodal complexes has grown in recent decades and has been subject of numerous reports.1-11 The reasons for this interest include their relevance to model functions of metalloenzymes1-3 and their potential applications in catalysis.13-17 The ligand system used most in this category has been tren, the tripodal tetraamine N(CH2CH2NH2)3, and its derivatives.4 The bz3tren is a versatile tetradentate ligand, known to form stable complexes not only with transition metals5-11 including Cu2+, Zn2+ and Co2+ but also anion species.12 However, only few results on the d10 metal complexes with bz3tren have been reported by us10 and others.6,7 As a part of on going efforts, we therefore focus our attention to extend other d10 system that includes heavy metal ions.

Mössbauer Study on the Variation in Magnetic Properties of CuO Induced by 57Fe Addition (57Fe 이온이 CuO에 미치는 효과에 관한 Mössbauer 분광 연구)

  • Park, Jae-Yun;Kim, Kwang-Joo
    • Journal of the Korean Magnetics Society
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    • v.19 no.3
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    • pp.113-119
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    • 2009
  • $^{57}Fe_xCu_{1-x}O$(x = 0.0, 0.02) powders were prepared by sol-gel method and their crystallographic and magnetic hyperfine properties have been studied using X-ray diffraction and $M{\ddot{o}}ssbauer$ spectroscopy (MS). The crystal structure of the samples is found to be monoclinic without any secondary phases and their lattice parameters increase with increasing annealing temperature ($T_A$), which is attributed to an increase in oxygen-vacancy content. MS measurements at room temperature indicate that $Fe^{3+}$ ions substitute $Cu^{2+}$ sites and ferromagnetic phase grow with increasing $T_A$. Magnetic hyperfine and quadrupole interactions of $^{57}Fe_{0.02}Cu_{0.98}O$ ($T_A=500^{\circ}C$) in the antiferromagnetic state at 17 K have been studied, yielding the following results: $H_{hf}=426.94\;kOe$, ${\Delta}E_Q=-3.67\;mm/s$, I.S.=0.32 mm/s, ${\theta}=65^{\circ}$, ${\phi}=0^{\circ}$, and ${\eta}=0.6$.

Studies on Heat Sensitivity of Egg Albumen II. Effects of pH and/or the Addition of Metal ions on Heat Sensitivity of Egg Albumen (난백의 숙감수성에 관한 연구 II. 금속염의 첨가와 pH가 난백의 열감수성에 미치는 영향)

  • 유익종;이성기;김영붕
    • Korean Journal of Poultry Science
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    • v.16 no.1
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    • pp.17-22
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    • 1989
  • In order to dull heat sensitivity of egg albumen, metal ions (aluminium, ferric, ferrous, copper) were added and functional properties or egg albumen were determined before and after heat treatment at $60^{\circ}C$ for 5 minutes. Effect of pH on heat sensitivity of aluminium salt added egg albumen was also determined. Addition of metal ions increased turbidity of egg albumen before and after the heat treatment. Changes of the turbidity were minimized by addition of aluminium salt. The foaming power was markedly increased by addition of ferric salt before the heat treatment and increased by addition of aluminium, ferric and copper salt after the heat treatment. Before the heat treatment the foam was stable by addition of ferric and ferrous salt but after the heat treatment it was stable by addition of aluminium and ferric salt. The turbidity and foaming property of the egg albumen with aluminium salt were not largely changed after the heat treatment at pH range 7 to 8.5. Over pH 9 the turbidity and foaming power were not decreased, but the foam stability was increased before and after the heat treatment. Salmonella typhimurium ATCC 14028 (10$^{6}$ cells/$m\ell$) inoculated in egg albumen at pH range 7 to 8.5 was destructed by the heat treatment.

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Selective catalytic reduction of NO by hydrocarbons over $Cu/Al_2O_3$ catalysts

  • Nam, Chang-Mo;Bernard M. Gibbs
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.4 no.4
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    • pp.201-208
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    • 2000
  • The reduction of NO by hydrocarbons was investigated over Cu/Al$_2$O$_3$catalysts using a stainless steel flow reactor under highly oxidising diesel exhaust conditions(up to 15%). Three different Cu loadings(1,5 and 10wt.%) on an $Al_2$O$_3$support were prepared and characterized using spectroscopic techniques. The catalytic activity tests show that different Cu loadings as well as temperature, oxygen, and hydrocarbon concentration levels significantly influence the NO reduction. Increasing Cu loadings up to 5 and 10wt.% decreases the catalytic activities for NO reduction due to the formation of a bulk crystalline CuO phase, as observed from XRD and SEM images. In particular, the visualization of the copper dispersion on the surface using the SEM-BEI technique provides information on the extent of copper saturation, particle size, and the effects on NO reduction. However, the lower Cu loading(1 wt.%) increases the catalytic activity with a temperature window of 720-810K, thereby favoring the formation of well dispersed isolated Cu species, e.g. Cu(sup)2+ ions, which is related to selective NO reduction. The effects of other reaction parameters, such as oxygen, the hydrocarbon level and type, and byproduct emissions are further discussed.

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Preferential Flow as Tested by Breakthrough Curves of Cl- and Cu2+ from Saturated Undisturbed Soil Core Samples under Steady Flow Conditions (포화 불교란 토양시료의 Cl- 및 Cu2+ 출현곡선에 의한 preferential flow의 검증)

  • Yoo, Sun-Ho;Han, Kyung-Hwa;Ro, Hee-Myong;Han, Gwang-Hyun
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.2
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    • pp.71-78
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    • 2000
  • Preferential flow has recently been the subject of increasing interest because these phenomena contribute to solute transport in soils. Commonly, preferential flow paths are associated with macropores or highly structured soils. We presented an analysis of the measured breakthrough curves (BTCs) of $Cl^-$ and $Cu^{2+}$ ions to test the occurrence of preferential flow in soils using miscible displacement technique under steady flow conditions. We also analyzed soil water retention curves and from this curves induced cumulative pore size distribution of undisturbed soils, which sampled from Ap1, B1, and C horizons of Songjeong series soils (the fine loamy, mesic family of Typic Hapludults). In this study, miscible displacement experiment on C horizon was excluded, because it is structureless sandy loam with saturated hydraulic conductivity of $5.2cmhr^{-1}$. The saturated hydraulic conductivity of Ap1 horizon was $2.0cmhr^{-1}$, which was about 7 times higher than that of B1 horizon ($0.27cm hr^{-1}$). Cumulative pore size distribution predicted that Ap1 horizon had more macropores (pore diameter larger than $49{\mu}m$, equivalent to -6 kpa of soil matric potential) than B1 horizon. The hydrodynamic dispersion coefficient from chloride BTCs was estimated as $1.3cm^2hr^{-1}$ for B1 and $34cm^2hr^{-1}$ for Ap1 horizon. However the retardation factors of B1 and Ap1 horizon were significantly different, i.e. 1 and 0.6, respectively, which means that there was distinct partition between mobile water and immobile phase in Ap1 horizon. The copper retardation effect of Ap1 horizon was less than that of B1 horizon, even though cation exchange capacity of Ap1 horizon was higher than that of B1 horizon. Thus, breakthrough curves of $Cl^-$ and $Cu^{2+}$ obviously showed the probability that preferential flow would occur in Ap1 horizon.

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Effect of Basic Oxygen Furnace Slag used as Structural Filling Materials on the Subsurface Environment (성·복토용으로 사용된 전로제강슬래그가 주변 토양환경에 미치는 영향)

  • Lee, Hosub;Nam, Taekwoo;Jho, Eun Hea;Nam, Kyoungphile
    • Journal of Soil and Groundwater Environment
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    • v.21 no.3
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    • pp.6-13
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    • 2016
  • The effect of blast oxygen furnace (BOF) slag used as filling materials on the soil environment was studied using column tests that simulated the flow of the BOF slag leachate through the soil layer. The Cu, Mn, Zn, Ni, and F contents of the leachate affected soil were similar to that of the controls (i.e., soils that were not affected by the leachate). The As, Cd, and Pb contents were lower in the leachate affected soils than the controls. The changes in these contaminants contents can be attributed to the interactions between anions such as alkalinity generating anions (e.g., CO32−, HCO3, OH) or calcium ions with heavy metals or F, which consequently affected the fate of heavy metals and F in the leachate affected soils. The germination and growth of Spinapis alba in the soils affected by the leachate and the controls were also similar. However, the proportion of alkalophilic bacteria in the soils affected by the leachate significantly increased, and this can be explained by the increased soil pH due to the alkaline leachate. Overall, this study shows that the alkalinity of the BOF slag leachate, rather than the presence of heavy metals and F in the leachate, needs to be considered when the BOF slag is to be reused as structural filling materials.

Fabrication of 3D Paper-based Analytical Device Using Double-Sided Imprinting Method for Metal Ion Detection (양면 인쇄법을 이용한 중금속 검출용 3D 종이 기반 분석장치 제작)

  • Jinsol, Choi;Heon-Ho, Jeong
    • Clean Technology
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    • v.28 no.4
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    • pp.323-330
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    • 2022
  • Microfluidic paper-based analytical devices (μPADs) have recently been in the spotlight for their applicability in point-of-care diagnostics and environmental material detection. This study presents a double-sided printing method for fabricating 3D-μPADs, providing simple and cost effective metal ion detection. The design of the 3D-μPAD was made into an acryl stamp by laser cutting and then coating it with a thin layer of PDMS using the spin-coating method. This fabricated stamp was used to form the 3D structure of the hydrophobic barrier through a double-sided contact printing method. The fabrication of the 3D hydrophobic barrier within a single sheet was optimized by controlling the spin-coating rate, reagent ratio and contacting time. The optimal conditions were found by analyzing the area change of the PDMS hydrophobic barrier and hydrophilic channel using ink with chromatography paper. Using the fabricated 3D-μPAD under optimized conditions, Ni2+, Cu2+, Hg2+, and pH were detected at different concentrations and displayed with color intensity in grayscale for quantitative analysis using ImageJ. This study demonstrated that a 3D-μPAD biosensor can be applied to detect metal ions without special analysis equipment. This 3D-μPAD provides a highly portable and rapid on-site monitoring platform for detecting multiple heavy metal ions with extremely high repeatability, which is useful for resource-limited areas and developing countries.

Flotation-Spectrophotometric Determination of Ag(I) at the 10-7 mol L-1 Level Using Iodide and Ferroin as an Ion-associate

  • Hosseini, Mohammad Saeid;Hashemi-Moghaddam, Hamid
    • Bulletin of the Korean Chemical Society
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    • v.26 no.10
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    • pp.1529-1532
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    • 2005
  • A simple and cost effective method for separation and preconcentration of Ag(I) at the $10^{-7}\;mol\;L^{-1}$ level in the environmental and mineral samples is present. The method is based on the flotation of Ag(I)-iodide complex as an ion-associate with ferroin in pH of 4 from a large volume of an aqueous solution (500 mL) using nheptane. The floated layer was then dissolved in dimethylsulfoxide (DMSO) for the subsequent spectrophotometric determination. Beer's law was obeyed over a range of 2.0 ${\times}$ $10^{-7}$-4.0 ${\times}$ $10^{-6}$ mol $L^{-1}$ with the apparent molar absorptivity of 2.67 ${\times}$ $10^5$ L $mol^{-1}\;cm^{-1}$. The detection limit (n = 5) was 4 ${\times}$ $10^{-8}$ mol $L^{-1}$, and RSD (n = 5) obtained for 2.0 ${\times}$ $10^{-6}$ mol $L^{-1}$ of Ag(I) was 2.2%. The interference effects of a number of elements was studied and found that only $Hg^{2+}$ at low concentration, and $Pb^{2+}$, $Cd^{2+}$, $Cu^{2+}$, and $Fe^{3+}$ ions at moderately high concentrations were interfered. To overcome on these interference effects, the solution was treated with EDTA at a buffering pH of 4 and passed through a column containing Amberlite IR-120 ionexchanger resin, just before the flotation process. The proposed method was applied to determine of Ag(I) in a synthetic waste water, a photographic washing sample and a geological sample and the results was compared with those obtained from the flame atomic absorption spectrometry. The results were satisfactorily comparable with together, so that the applicability of the proposed method was confirmed in encountering with the real samples.