• Korean Journal of Biological Psychiatry
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• v.5 no.2
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• pp.235-242
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• 1998

Objectives : Alcohol-induced oxidative stress has been known to injure various tissues or organs. This stress is related with free radicals which are produced as the result of long-term alcohol consumption. Malonyldialdehyde(MDA) is produced by the interaction of free radicals and cell membrane lipids, and indicates the degree of lipid peroxidation indirectly. The purpose of this study was to investigate the relationship between red blood cell(RBC) membrane lipid peroxidation by free radicals, and associated hepatic injuries and hematologic changes. Methods : Thirty-three subjects diagnosed as alcohol dependence according to DSM-IV diagnostic criteria were evaluated within 72 hours after discontinuing alcohol drinking. Clinical characteristics were evaluated by CAGE questionnaire and Korean Michigan Alcoholism Screening Test(MAST). RBC membrane MDA level was measured as the marker of RBC membrane lipid peroxidation. Aspartate aminotransferase(AST), alanine aminotransferase(ALT) and gamma-glutamyltransferase(GGT) were used as the biochemical markers of liver damage due to alcohol ingestion. The alcohol-induced hematologic change was assessed by mean corpuscular volume(MCV). Results : The results were as follows. Clinical characteristics were not different between two groups having normal and abnormal levels of AST, ALT, GGT or MCV. The levels of MDA were not correlated with the clinical characteristics and serum levels of AST, ALT and GGT. However, there was a significant correlation between the levels of MDA and the value of MCV(p=0.017). Conclusions : These findings suggest that oxidative stress in alcohol dependence may not be reflected in liver enzyme markers such as AST, ALT and GGT, but may be reflected in MCV.

• Korean Journal of Veterinary Research
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• v.35 no.4
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• pp.713-727
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• 1995

The present study was investigated fatty acid composition of the phospholipid in the RBC membrane, liver and microsomal fraction after vitamin E and/or insulin treatment to evaluate the effect of vitamin E on the oxidative stress in STZ-treated rat and BB rat. Results obtained through the experiments were summarized as follows; 1. Effect of vitamin E and/or insulin treatment in STZ-treated rat 1) In the insulin treated group and the combination treated groups of vitamin E with insulin, body weights were increased compared to STZ-treated rat(STZ control group). Especially it was more significantly increased in the combination treated group of high dose vitamin E with insulin. 2) The composition of fatty acids of the phospholipid in RBC membrane, liver and microsomal fractions was shown a decreased C16:1, C18:1, C20:4 and an increased C16:0, C18:0, C18:2 in STZ control group compared to normal control group. In RBC membrane, liver and microsomal fractions after vitamin E with insulin treatment in STZ-treated rat, effect on the composition of fatty acids of the phospholipid was shown the result of a decreased C16:0, C18:0, C18:2 and an increased C16:1, C18:1, C20:4. 3) Hemolysis rate of the RBC to $H_2O_2$ was increased in the STZ control group and it was decreased below the hemolysis level of normal control group by vitamin E treatment. 2. Effect of vitamin E and/or insulin treatment in BB rat 4) Only in microsomal fraction, fatty acid composition was different between insulin treatment group and vitamin E with insulin treatment group. It was increased C16:0 and C18:1, and decreased C18:0 and C18:2 in vitamin E with insulin treatment group: But C20:4 was not different in two groups. These results suggest that the combination treatment of vitamin E and insulin could prevent the oxidative change of fatty acids in P-lipid of the RBC membrane, liver and microsomal fraction in STZ-treated rats and BB rats.

• The Korean Journal of Physiology
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• v.23 no.1
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• pp.13-21
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• 1989

The present study was designed to investigate i) the action of various nucleotides on membrane permeability of rat red blood cell and hepatocyte for $Na^{+}$ and $Rb^{+}$ ii) the characteristics of purinoceptors on these cell membranes. Blood from Sprague-Dawley rats was obtained by carotid arterial cannulation. Red blood cells were then washed 3 times with saline at $4{\circ}C$. Hepatic parenchymal cells were isolated from rat livers by using a modification of the Berry and Friend (1969) method. For the $Na^{+}$ influx studies, isolated RBC and hepatocyte were incubated in incubation medium containing $^{22}Na^{+}0.2\;{\mu}Ci/ml$ at $37^{\circ}C$. After various time intervals samples were removed from the incubation flask and washed out 3 times with ice-cold washing solutions. Cells were destroyed by adding Triton X-100 and TCA solution. After centrifugation, the supernatants were assayed for $^{22}Na^{+}$ by gamma counter. $^{86}Rb^{+}$ was used to simulate $K^{+}$ in these $K^{+}efflux$ studies. Isolated hepatocytes were incubated for 60 min in the loading solution containing $^{86}Rb^{+}\;10\;{\mu}Ci/ml$ at $37^{\circ}C$. After loading, the cells washed out 3 times by centrifugation with washing solution. The cells were incubated in buffer solution at $37^{\circ}C$. At intervals thereafter, samples were removed and centrifuged. The supernatants were analyzed for $^{86}Rb^{+}$ by liquid scintillation counter. The main results of the experiments were: 1) ATP and ATPP increased in both $^{22}Na^{+}$ influx and $^{86}Rb^{+}$ efflux in the red blood cell. Although ADP showed a tendency to increase in RBC membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$, the changes were not significantly different from the control. 2) The Significant changes in $^{22}Na^{+}$ and $^{86}Rb^{+}$ flux by ATP were also demonstrated in hepatocyte. ATPP and ADP showed a tendency to increase in hepatocyte membrane permeability for both ions. 3) Other nucleoside triphosphates-ITP, GTP and CTP-did not change in membrane permeability for $^{22}Na^{+}$ and $^{86}Rb^{+}$ in RBC and hepatocyte. In conclusion, not only ATP but also ATPP activate purinoceptors and change in membrane permeability for $Na^{+}$ and $K^{+}$. In order to activate purinoceptors on the cell membrane, the nucleotides have to possess intact adenine moiety and three phosphates or more in its molecule.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.1
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• pp.39-47
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• 2009

Gaegurin 4(GGN 4), an antimicrobial peptide isolated from a Korean frog, is five times more potent against Gram-positive than Gram-negative bacteria, but has little hemolytic activity. To understand the mechanism of such cell selectivity, we examined GGN4-induced $K^+$ efflux from target cells, and membrane conductances in planar lipid bilayers. The $K^+$ efflux from Gram-positive M. luteus(2.5 ${\mu}g/ml$) was faster and larger than that from Gram-negative E. coli(75 ${\mu}g/ml$), while that from RBC was negligible even at higher concentration(100 ${\mu}g/ml$). GGN4 induced larger conductances in the planar bilayers which were formed with lipids extracted from Gram-positive B. subtilis than in those from E. coli(p<0.01), however, the effects of GGN4 were not selective in the bilayers formed with lipids from E. coli and red blood cells. Addition of an acidic phospholipid, phosphatidylserine to planar bilayers increased the GGN4-induced membrane conductance(p<0.05), but addition of phosphatidylcholine or cholesterol reduced it(p<0.05). Transmission electron microscopy revealed that GGN4 induced pore-like damages in M. luteus and dis-layering damages on the outer wall of E. coli. Taken together, the present results indicate that the selectivity of GGN4 toward Gram-positive over Gram-negative bacteria is due to negative surface charges, and interaction of GGN4 with outer walls. The selectivity toward bacteria over RBC is due to the presence of phosphatidylcholine and cholesterol, and the trans-bilayer lipid asymmetry in RBC. The results suggest that design of selective antimicrobial peptides should be based on the composition and topology of membrane lipids in the target cells.

• Journal of Chest Surgery
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• v.25 no.7
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• pp.693-701
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• 1992

Extracorporeal cardiopulmonary bypass[CPB] has been associated with a wide variety of hematologic derangements, including a transient deformation and hemolysis of red blood cells[RBCs], which is supposed to be due to mechanical trauma and/or metabolic alterations. Since membrane integrity is, in part, maintained by energy requiring process, inadequate function of erythrocyte glycolytic pathway, which is inevitalble during CPB, may cause depletion of high energy phosphate pool and result in hemolysis. The authors performed an investigation to assess whether administration of Fructose-l, 6-diphsphate [FDP], which has been known to enhance intracellular glycolytic activities, could counteract erythrocyte hemolytic events caused by CPB. Sixty pateints with cyanotic congenital heart diseases, who underwent open heart surgery under CPB longer than 60 minutes, were randomly divided into two groups depending on whether use of FDP[Group FDP] or not[Group Control]. The age, sex, CPB time, preoperative hemoglobin level, disease entities were all similar[Table 1], and membrane type oxygenators were used in all patients. In Group, FDP, a dose of 250mg/kg body weight of FDP was administered by intravenous dripping every 12 hours from the morning of the operation to postoperative 48 hours, To demonstrate the degree and pattern of hemolysis of erythrocyte, reticulocyte count, indirect /direct bilirubin, haptoglobin, plasma hemoglobin, lactate dehydrogenase were measured every 12 hours from the time of cessation of CPB to 48 hours and RBC morphologic study, osmotic fragility test were done every 24 hours. All parameters revealed less hemolytic in group FDP [Fig. 1~5], though the differences between two groups were not significant, except plasma hemoglobin, lactate dehydrogenase changes. A pattern of sequential changes of plasma hemoglobin, lactate deh-ydrogenase showed the highest level at the time of CPB stop and abrupt decrease in following 24 hours in both groups, and statistically significant differences were demonstrated in group FDP at least for the first 12 hours postoperatively[p<0.05]. The authors conclude that they can expect the benificial effect of FDP on the maintenance of membrane stability of RBC probably by energy enhancement during the shock status of CPB, but FDP could not completely prevent the damaging effect on RBC by cardiopulmonary bypass

• Proceedings of the Korean Magnestics Society Conference
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• 2015.05a
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• pp.161-163
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• 2015

The micro device, coil, and channel for the biosensor integrated with the GMR-SV device based on the antiferromagnetic IrMn layer was fabricated by the light lithography process. When RBCs coupled with several magnetic beads with a diameter of $1{\mu}m$ passed on the micro channel, the movement of RBC + ${\mu}Beads$ is controlled by the electrical AC input signal. The RBC + ${\mu}Beads$ having a micro-magnetic field captured above the GMR-SV device is changed as the output signals for detection status. From these results, the GMR-SV device having the width magnitude of a few micron size can be applied as the biosensor for the analysis of a new magnetic property as the membrane's deformation of RBC coupled to magnetic beads.

• The Korean Journal of Zoology
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• v.37 no.4
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• pp.597-604
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• 1994

Membranes obtained from the normal human RBC population were separated by continuous sucrose density gradient centrifugation and the density-fractionated membranes were then examined for changes in molecular markers. This study focuses on changes of (i) the membrane protein profile, (ii) differences in membrane-associsted enzyme activities, and (iii) the amount of autologous IgG bound. The following observations were made: (i) ratios for band 4. la over the sum of bands (4. la + 4.Ib) ranged from 0.58 to 0.79 for membranes of lowest density; (ii) significant changes in bound glyceraldehyde-3-phosphate dehydrogenase and acetvlcholinesterase activities were found; (iii) the amounts of autolosous IgG's attached to the red blood cells was highest in the membrane fraction of lowest density.

• Proceedings of the Korean Vacuum Society Conference
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• 2016.02a
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• pp.208.2-208.2
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• 2016

Many researchers have paid attention to the studies on the interaction between non-thermal plasma and aqueous solutions for biomedical applications. The gas composition in the plasma is very important. Oxygen and nitrogen are the main gases of interest in biological applications. Especially, we focus on the oxygen concentration. In this experiment, we studied the role of oxygen concentration in plasma induced chemical reactions in solution. At first, the amount of ions are measured according to changing the oxygen concentration. And we checked the relationship between these ions and pH value. Secondly, when the oxygen concentration is changed, it identified the type and amount of radical generated by the plasma. In order to confirm the effect of these chemical property change to biological material, hemoglobin and RBCs are chosen. RBCs are one of the common basic biological cells. Thirdly, when plasma treated according to oxygen concentration in nitrogen feeding gas, oxidation of hemoglobin and RBC is checked. Finally, membrane oxidation of RBC is measured to examine the relation between hemoglobin oxidation and membrane damage through relative hemolysis and Young's modulus. Our results suggest that reactive species generated by the plasma differsdepending on the oxygen concentration changes. The pH values are decreased when oxygen concentration increased. OH decrease and NO increase are also observed. These reactive species makes change of chemical properties of solution. We also able to confirm that the difference in these reactive species to affect the oxidation of the Hb and RBCs. The Hb and RBCs are more oxidized with the high oxygen concentration conditions. But membrane is damaged more by plasma treatment with only nitrogen gas. It is shown that red blood cells membrane damage and oxidation of hemoglobin are not directly related.

• Reproductive and Developmental Biology
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• v.36 no.1
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• pp.55-64
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• 2012

Peroxiredoxin II (Prdx II; a typical 2-Cys Prdx) has been originally isolated from erythrocytes, and its structure and peroxidase activity have been adequately studied. Prdx II has been reported to protect a wide range of cellular environments as antioxidant enzyme, and its dysfunctions may be implicated in a variety of disease states associated with oxidative stress, including cancer and aging-associated pathologies. But, the precise mechanism is still obscure in various aspects of aging containing ovarian aging. Identification and relative quantification of the increased proteins affected by Prdx II deficiency may help identify novel signaling mechanisms that are important for oxidative stress-related diseases. To identify the increased proteins in Prdx $II^{-/-}$ mice, we performed RBC comparative proteome analysis in membrane fraction and cytosolic fractions by nano-UPLC-$MS^E$ shotgun proteomics. We found the increased 86 proteins in membrane (32 proteins) and cytosolic (54 proteins) fractions, and analyzed comparative expression pattern in healthy RBCs of Prdx $II^{+/+}$ mice, healthy RBCs of Prdx $II^{-/-}$ mice, and abnormal RBCs of Prdx $II^{-/-}$ mice. These proteins belonged to cellular functions related with RBC lifespan maintain, such as cellular morphology and assembly, cell-cell interaction, metabolism, and stress-induced signaling. Moreover, protein networks among the increased proteins were analyzed to associate with various diseases. Taken together, RBC proteome may provide clues to understand the clue about redox-imbalanced diseases.

• Proceedings of the KSME Conference
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• 2004.11a
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• pp.1505-1509
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• 2004

The suspension of hardened red blood cells (RBCs) differs from the suspension of normal RBCs with respect to their rheological behavior. The deformability of normal and hardened RBCs (obtained by heating blood at $49^{\circ}C$ or by incubating RBCs in a solution of hydrogen peroxide) was measured with a slit diffractometer and RBC suspension viscosity was measured with a rotational viscometer. The peroxide-treated RBCs showed a significant decrease of the deformability and their suspension viscosity increased over a range of shear rates. The suspension viscosity of the heated RBCs, however, where the deformability is even lower than that of the peroxide-treated RBCs, was slightly higher than that of the normal RBC suspension in the high shear rates. The present study found that not all rigid cells cause an increase of blood viscosity at high shear rate, and therefore that decreased membrane deformability is not predictive of high-shear blood viscosity.