• 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.

• The Korean Journal of Physiology
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• v.20 no.2
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• pp.218-224
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• 1986

The characteristics of anion exchange with internal $HCO_3\;^{-}\;(or\;OH^-)$ was studied by determining the time course of hemolysis in isoosmotic ammonium salt solution in human erythrocytes. The effects of inhibitors, pH and temperature on the exchange between internal $HCO_3\;^-\;(or\;OH^-)$ and external $Cl^-$ were observed and the permeabilities of various organic and inorganic anions were also measured. The results were compared with data previously reported from the experiments using radioisotopes. The results are as follows; 1) SITS $H_2DIDS$ and furosemide inhibited the hemolysis of erythrocytes in isoosmotic $NH_4Cl$ solution in a dose·dependent manner, and the concentrations for lengthening twice the time for $half-hemloysis(t_{1/2})\;were\;2.3{\times}10^{-7},\;1.3{\times}10^{-7}\;and\;2.5{\times}10^{-5}M$, respectively. 2) Acetazolamide also shifted the time-dependent hemolytic curve to the right in a dose-dependent manner, and the concentrations for lengthening twice $t_{1/2}\;was\;2.4{\times}10^{-5}M$. 3) The time-dependent hemolysis was delayed by decreasing pH from 7.0 to 6.2, but w·as not affected by the change of pH in the range of 7.0 to 8.2. 4) The time for $half-hemloysis(t_{1/2})$ showed a temperature-dependency and Arrhenius plot exhibited a break point at $20^{\circ}C$. The apparent activation energy calculated from this plot was 18.1 kcal/mol between $2^{\circ}C-20^{\circ}C$ and 11.2 kcal/mol between $20^{\circ}C-37^{\circ}C$, respectively. 5) The apparent permeabilities of various inorganic anions based on $t_{1/2}$ were in the order of $Cl^->NO_{3}\;^->SCN^->SO_4\;^{2-}>SSO_3\;^{2-}>HPO_4\;^{2-}$. which was similar with the previous reports based on the experiment using radioisotopes. The results Obtained from this study are comparable with the previous data reported from the experiments using radioisotopes. This indicates that the hemolysis of erythrocytes in isoosmotic ammonium salt solution can be used as a simple and good method for the study of anion exchange in erythrocyte membrane.

• Journal of Environmental Science International
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• v.4 no.2
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• pp.223-232
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• 1995

New two macrocyclic compounds using as carriers of liquid emulsion menbrame, have been synthesized. These reuslts provide evidance for the usefulness of the theory in designing the systems. The efficiency of selective transport for heavy metal ions have been discussed from the membrane systems that make use of $SCN^-$,<>,$I^-$,CN- and $Cl^-$ ion as co-anions in source phase and make use of $S_2O_3^{2-}$ and $P_2O_7^{4-}$ ion as receiving phase, respectively. The transport rate of M(II) was highest when a maximum amount of the M(II) in the source phase was present as$Cd(SCN)_2$$(P[SCN^-]= 0.40M)$, $Hg(SCN)_2([SCN^-]=0.40M)$ and Pd(CN)$([CN^-]= 0.40M)$. The Cd(II) and Pb(II) over each competitive cations were well transprted with 0.3M-S2032- and 0.3M-P2O74-, respectively in the receiving phase. Results of this study indicate that two criteria must be met in order to have effective macrocycle-mediated transport in these emulsion system. First one must effective extraction of the $M^{n+}$ into the toluene systems. The effectiveness of this extraction is the greatest if locK for $M^{n+}$macrocycle interaction is large and if the macrocycle is very insoluble in the aqueous phase. Second, the ratio of the locK values (or Mn+-receiving phase ($S_2O_3^{2-}$- or $P_2O_7^{4-}$) to $M^{n+}$-macrocycle (($L_1$이나 $L_2$) interaction must be large enough to ensure quantitative stripping of Mn+(($Cd^{2+}$,$Pb^{2+}$)at the toluene receiving Phase interface. $L_1$(3.5-benzo-10,13,18,21-tetraoxa-1,7,diazabicyclo(8,5,5) eicosan) forms a stable ($Cd^{2+}$ and >,$Pb^{2+}$ complexes and $L_1$ is very insoluble in water and its $Cd^{2+}$ and >,$Pb^{2+}$ complex is considerably less stable than $Cd^{2+}$-(S2O3)22- and $Pd^{2+}-P_2O_7^{4-}$ complexes. On the other hand, the stability of the $Hg^{2+}$)+-$L_1$( complex exceed that of the $Hg^{2+}$- (S2O3)22- and Hg2+-P2O74-, and the distribution coefficient of $L_2$(5,8,15,18,23,26-hexaoxa-1,12- diazabicyclo-(10,8,8) octacosane) is much smaller than that of $L_1$. Therefore, the partitioning of Lr is favored by the aqueous receiving Phase, and little heavy metal ions transport is seen despite the large logK for $Hg^{2+}$+-$L_1$ and $Mn^+$($Cd^{2+}$+, $Pb^{2+}$+ and $Hg^{2+}$)-$L_2$ interactions. Key Words : macrocycles, transport, heavy metal, co-anion, source phase, receiveing, complex separation, interaction, destribution coefficient.

• Journal of Environmental Science International
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• v.18 no.6
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• pp.645-654
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• 2009

Lab-scale Electrodialysis(ED) system with different membranes combined with before or after pyroma process were carried out to remove nitrate from two pickling acid wastewater containing high concentrations of $NO_3\;^-$(${\approx}$150,000 mg/L) and F($({\approx}$ 160,000 mg/L) and some heavy metals(Fe, Ti, and Cr). The ED system before Pyroma process(Sample A) was not successful in $NO_3\;^-$ removal due to cation membrane fouling by the heavy metals, whereas, in the ED system after Pyroma process(Sample B), about 98% of nitrate was removed because of relatively low $NO_3\;^-$ concentration (about 30,000 mg/L) and no heavy metals. Mono-selective membranes(CIMS/ACS) in ED system have no selectivity for nitrate compared to divalent-selective membranes(CMX/AMX). The operation time for nitrate removal time decreased with increasing the applied voltage from 10V to 15V with no difference in the nitrate removal rate between both voltages. Nitrate adsorption of a strong-base anion exchange resin of $Cl\;^-$ type was also conducted. The Freundlich model($R^2$ > 0.996) was fitted better than Langmuir mode($R^2$ > 0.984) to the adsorption data. The maximum adsorption capacity ($Q^0$) was 492 mg/g for Sample A and 111 mg/g for Sample B due to the difference in initial nitrate concentrations between the two wastewater samples. In the regeneration of ion exchange resins, the nitrate removal rate in the pickling acid wastewater decreased as the adsorption step was repeated because certain amount of adsorbed $NO_3\;^-$ remained in the resins in spite of several desorption steps for regeneration. In conclusion, the optimum system configuration to treat pickling acid wastewater from stainless-steel industry is the multi-processes of the Pyroma-Electrodialysis-Ion exchange.

• Resources Recycling
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• v.24 no.5
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• pp.33-39
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• 2015

A basic study was conducted to effectively recover nitric acid from a waste solder stripper by diffusion dialysis using anion exchange membranes. The effects of flow rate, flux ratio, nitrate concentration, and metallic ion types and concentration on the recovery percentage of nitric acid were investigated. The recovery percentage of nitric acid was decreased with the increase of flow velocity. But the recovery percentage of nitric acid was increased as the increase of flux ratio(W/F) and showing a recovery percentage of nitric acid of about 99% at a flux ratio of 1.5 or more. As the increase of nitric acid concentration in feed solution, the recovery percentage of nitric acid was increased up to 3.0M, but in case of greater than 3.0M, the recovery percentage gradually was decreased. Leakage percentage of metallic ions through the membrane were in the order of Pb, Na and Cu but Fe and Sn did not leakaged. As a result of diffusion dialysis using real waste solder stripper at a flow rate of $0.9L/hr-m^2$, W/F = 1.3, a recovery percentage of nitric acid of approximately 94% was gained.

• Resources Recycling
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• v.33 no.4
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• pp.36-46
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• 2024

We investigated a hydrometallurgical process of nickel recovery from Inconel 713C scrap. The process proceeded with a series of i) comminution of pyrometallurgical treated scrap, ii) sulfuric acid leaching, iii) solvent extraction of unreacted acid, molybdenum, aluminum, and precipitation of chromium, iv) crystallization of nickel sulfate by vacuum evaporation, and v) nickel electrowinning. The nickel-aluminum intermetallic compound, Ni₂Al₃, was formed by the pyrometallurgical pretreatment readily grounded under 75 ㎛. Sulfuric acid leaching was done for 2 hours in 2 mol/L, 20 g/L solid/liquid ratio, and 80 ℃. It revealed that over 98 % of nickel and aluminum was dissolved, whereas 28 % of molybdenum was. A nickel sulfate solution with 2.34 g/L for the crystallization of nickel sulfate hydrate was prepared via solvent extraction and precipitation. Over 99 % of molybdenum and aluminum and 93 % of chromium was removed. Nickel metal with 99.9 % purity was obtained by electrowinning with the nickel sulfate monohydrate in the cell equipped with anion exchange membranes for catholyte pH control. The membrane did not work well, resulting in a low current efficiency of 73.3 %.

• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.305-312
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• 1996

Cell volume regulatory mechanisms are usually disclosed by exposure of cell to anisotonic media. If a cell is suddenly exposed to hypotonic media, it swells initially like an osmometer but within minutes regains its original cell volume. This behavior has been labelled as regulatory cell volume decrease(RVD). RVD is believed to result from the loss of permeable ions through the membrane. In this study, we examined hypotonically induced changes in the membrance currents involved in RVD by using whole cell voltage clamp technique in the unfertilized hamster egg. At -40mV of the holding potential, the stationary current was maintained in the hamster egg exposed to isotonic solution composed of, mainly, 115mM NaCl and 40mM mannitol. Hypotonic solution was prepared by removing mannitol. Therefore, the concentrations of $Na^+$ and $Cl^-$ in this hypotonic media were the same as those in the isotonic solution. Following 30 to 60 sec after applying the hypotonic media to the egg, the inward current was evoked. This inward current was eliminated by $100{\mu}M$ 4-acetamido-4'-isothiocyanostil-bene-2,2'-disulfonic acid(SITS), an anion channel blocker, leaving the small outward current component. Further addition of 2mM $Ba^{2+}$, a broad $K^+$ channel blocker, completely abolished the small outward current left even in the presence of SITS during hypotonic stress. These results suggest that $K^+$ and $Cl^-$ move out of cells, resulting in RVD. To test the involvement of $Na^+$ in RVD, 20mM Na-isethionate was substituted for mannitol in isotonic media(135mM $Na^+$) and Na-isethionate (20mM) was freed the hypotonic solution. Only $Cl^-$ concentration in both isotonic and hypotonic media was kept constant at 115mM, whereas concentration of $Na^+$ was lowered in hypotonic solution to 115mM from 135mM in isotonic solution. Hypotonic medium induced the outward current in the egg equilibrated isotonically. This current was reduced by $100{\mu}M$ SITS but was augmented by 2 mM $Ba^{2+}$. In terms of RVD, these results imply that $Cl^-$ efflux is coupled with $K^+$, maybe for electroneutrality during hypotonic stress and/or with $Na^+$ via unknown transport mechanism(s). From the overall results, the hypotonic stress facilitates the movement of $Cl^-$ and $K^+$ out of the hamster egg to regain cellular volume with electroneutrality. If there exist a difference in $[Na^+]_0$ between isotonic and hypotonic solution, another transport mechanism concerned with $Na^+$ may, at least partly, participate in regulatory volume decrease.