• Journal of Physiology & Pathology in Korean Medicine
• /
• v.27 no.5
• /
• pp.631-636
• /
• 2013

This study was to evaluate the antioxidant properties of the leaf extracts of Diospyros lotus (DLE) on the chemical-induced free radical and rat red blood cell (RBC) oxidative damage in vitro. DLE were prepared by extracting with water. DLE showed the high antioxidant activities on the scavenging of 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid)-induced radicals. An antioxidant activities of DLE was similar to the reference antioxidant butylated hydroxytoluene (BHT) and (${\pm}$)6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid (Trolox). Reducing power of $1,000{\mu}g/mL$ DLE also was similar to the vitamin C. In RBC, oxidative hemolysis induced by the aqueous peroxyl radical generator (2,2'-Azobis (2-methylpropionamidine) dihydrochloride (AAPH)) were significantly suppressed by DLE in a dose-dependent manner. Furthermore, DLE prevented the depletion of cytosolic antioxidant glutathione in RBC damaged with AAPH. These results suggest that DLE may has value as natural product with its high quality antioxidant properties against oxidative stress.

• Clinical and Experimental Pediatrics
• /
• v.59 no.2
• /
• pp.100-103
• /
• 2016

Patients with hemolytic-uremic syndrome (HUS) can rapidly develop profound anemia as the disease progresses, as a consequence of red blood cell (RBC) hemolysis and inadequate erythropoietin synthesis. Therefore, RBC transfusion should be considered in HUS patients with severe anemia to avoid cardiac or pulmonary complications. Most patients who are Jehovah's Witnesses refuse blood transfusion, even in the face of life-threatening medical conditions due to their religious convictions. These patients require management alternatives to blood transfusions. Erythropoietin is a glycopeptide that enhances endogenous erythropoiesis in the bone marrow. With the availability of recombinant human erythropoietin (rHuEPO), several authors have reported its successful use in patients refusing blood transfusion. However, the optimal dose and duration of treatment with rHuEPO are not established. We report a case of a 2-year-old boy with diarrhea-associated HUS whose family members are Jehovah's Witnesses. He had severe anemia with acute kidney injury. His lowest hemoglobin level was 3.6 g/dL, but his parents refused treatment with packed RBC transfusion due to their religious beliefs. Therefore, we treated him with high-dose rHuEPO (300 IU/kg/day) as well as folic acid, vitamin B12, and intravenous iron. The hemoglobin level increased steadily to 7.4 g/dL after 10 days of treatment and his renal function improved without any complications. To our knowledge, this is the first case of successful rHuEPO treatment in a Jehovah's Witness child with severe anemia due to HUS.

• Journal of Nutrition and Health
• /
• v.29 no.2
• /
• pp.232-241
• /
• 1996

This study was designed to compare the effect of different dietary fats on plasma lipids, the degree of lipid peroxidation and the activity of antioxidant enzymes in RBC and liver rats treated with or wighout 1, 2-dimethylhydrazing (DMH). Male Sprague Dawley rats, at 7 weeks-old, were divided into control and DMH-treated grous, and each group was again subdivided into four were perilla oil (PO), blend fat (BF) containing ten different kinds of dietary oil, beef tallow (BT), corn oil (CO). At the same time, each rat was injected intramusculary with saline(for control) or DMH twice a week for 6 weeks to give total dose of 180 mg/kg body weight. Compared with BT feeding, BF reduced plasma total choesterol level and PO and Co reduced plasma TG levels (p<0.05). DMH injection decreased plasma cholesterol in all dietary groups. However, PO decreased tocopherol levels and increased TBARS levels in RBC compared to BT. The degree of hemolysis in PO group was higher than that of BT group (p<0.05 only in control group. Fatty acid composition of hepatic microsome was reflected by dietary fatty acid profile. The peroxidizability index and TBARS level in hepatic micorsome were significantly increased but tocopherol level was lowered in PO group compared to BT group. Activites of superoxide dismutase and glutathione peroxidase in RBC and hepatic cytosol were not influenced y dietary fats and DMH treatment(p<0.05). Overall, perilla oil rich in $\omega$3 $\alpha$-linolenic acid could be a very important dietary source in reducing plasma lipids and blend fat was also good dietary oil mixture in reducing plasma cholesterol. However, the degree of lipid peroxidation was greater in tissue by perilla oil feeding and it is very difficult to use only perilla oil as oil source for meal preparation, so that it could be suggested to use more perilla oil and fish to give an equal effect of blend fat in order to reduce the risk factors against cardiovascular disease.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
• /
• v.12 no.2
• /
• pp.20-52
• /
• 1999

In order to investigate the effect of Naetakchungumsankamibang(NTCGS) water extract on the skin tumor induced by 3-MCA and immunological responses in mice, the cytotoxicity against SK-MEL-2 cells and total number of tumors induced by 3-MCA were measured. The numbers of WBC, platelets and RBC, plaque forming cells, hemagglutinin titer, hemolysis titer, carbon clearance, proliferation of splenocyte by thymidine uptake assay, splenic leukocyte by FACS analysis and $TNF-{\alpha}$ were also measured for the evaluation of the immunological responses. The results were obtained as follows: 1. In cytotoxicity against SK-MEL-2 cells, concentration inhibiting cell growth up to below $20\%$ of control was recognized at 1mg/ml of NTCGS. 2. In Inhibitory effect on the skin tumor induced by 3-MCA, the results showed a strong inhibitory effect of NTCGS. 3. In hematological changes in the tumor bearing mice, the numbers of WBC decreased significantly in NTCGS treated group as compared with control. 4. In hematological changes in the tumor bearing mice, the numbers of platelets increased significantly in NTCGS treated group as compared with control. 5. In hematological changes in the tumor bearing mice, the numbers of RBC increased with no significance in NTCGS treated group as compared with control. 6. Effects of the plaque forming cells in the tumor bearing mice, NTCGS treated group exhibited a significant effect compared with control. 7. In terms of the effects on hemagglutinin titer, NTCGS treated group showed higher level than control, without significance. 8. In terms of the effects on hemolysis titer, NTCGS treated group showed higher level than control, without significance. 9. In terms of the effects on phagocytic index K in Balb/C mice, NTCGS treated group showed significant difference from control. 10. In terms of the effects on proliferation of splenocyte by thymidine uptake assay, NTCGS showed significant effect at the concentration of 0.5mg/ml. 11. In terms of the effects on splenic leukocyte of Balb/C mice by FACS analysis, NTCGS treated group showed significantly higher level of helper T cell, B cell and macrophage than in control. 12. In terms of the effects on the secretion of $TNF-{\alpha}$, the treated group showed significant effect at the concentration of 1mg/ml of NTCGS. Based on the results summarized above, NTCGS is considered to have antitumor activity and immunological responses against skin tumor, and to be usable fur the treatment.

• Journal of Oriental Neuropsychiatry
• /
• v.10 no.2
• /
• pp.71-83
• /
• 1999

This study was to investigate the effects of SUNGSIMJIHWANGTANG(SSJHT) on the blood and brain tissues of aged rats. For the experiment, the aged rats were divided into three groups : Non treated group(NC), distilled water fed group(PC), SSJHT fed group(SSJHT). Each group was fed for ten days before administration of scopolamine. Then, we injected scopolamine intraperitoneally to PC and SSJHT group for 5 days. We observed the changes of their blood cell(WBC, RBC, platelet), blood serum(BUN, creatinine, glucose, uric acid), erythrocyte hemolysis, and the activities of cholinesterase and measured the amounts of malondialdehyde in the serum, catalase, and SOD in the brain tissue. The main results of this investigation are as follows. 1. In respect of the number of WBC, SSJHT group exhibited significant increase in comparison with PC. 2. In respect of the amount of creatinine and uric acid in the blood serum, SSJHT group exhibited significant decrease in comparison with PC. 3. In respect of erythrocyte hemolysis, SSJHT group exhibited significant suppression in comparison with PC. 4. In respect of the activity of cholinesterase in the serum, SSJHT group exhibited significant activation in comparison with PC. 5. In respect of the amounts of malondialdehyde in the serum, SSJHT group exhibited significant decrease in comparison with PC. 6. In respect of the activity of catalase in brain tissue, SSJHT group didn't exhibit significant change in comparison with PC. 7. In respect of the activity of SOD in brain tissue, SSJHT group exhibited significant activation in comparison with PC. As a result of this study, SSJHT is expected to have antiaging effect by suppressing the formation of free radicals, the accumulation of antioxidants and further study needs to be carried on about SSJHT.

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

• Korean Journal of Pharmacognosy
• /
• v.43 no.1
• /
• pp.66-71
• /
• 2012

Cirsium japonicum var. ussuriense is often used in treatment of human disease such as hemorrhage, blood congestion and inflammation. This study was accomplished to evaluate the antioxidant properties of the leaf (CLE) and flower (CFE) extracts of C. japonicum var. ussuriense to protect normal human red blood cells (RBC) and plasma samples against oxidative damage in vitro. CLE and CFE were prepared by extracting with hot water. In red blood cells and plasma, oxidative hemolysis and lipid peroxidation induced by the aqueous peroxyl radical generator [2,2'-azobis (2-amidinopropane) dihydrochloride, AAPH] were significantly suppressed by CLE or CFE in a dose-dependent manner at the same time. CLE and CFE also prevented the depletion of cytosolic antioxidant glutathione (GSH) in RBC. These results suggest that the leaves and flowers of C. japonicum var. ussuriense may have the antioxidant properties.

• Journal of Applied Biological Chemistry
• /
• v.59 no.3
• /
• pp.221-225
• /
• 2016

Tolaasin is a pore-forming peptide toxin produced by Pseudomonas tolaasii and causes a brown blotch disease by disrupting membrane structures of cultivated mushrooms. The mechanism and characteristics of tolaasin pore formation are not known in detail; however, tolaasin pores have been demonstrated in the artificial lipid bilayer. Since the tolaasin pore appeared less frequently and unstable in lipid bilayer, a mismatch between the length of tolaasin pore and the thickness of lipid membrane had been suggested. Therefore, tolaasin-induced hemolyses were measured by the additions of phospholipids composed of various fatty acids with different carbon numbers. When phosphatidylethanolamines made with two decanoic acids (C10:0, 1,2-didecanoyl-sn-glycero-3-phosphoethanolamine; DDPE), myristic acids (C14:0, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine), and stearic acids (C18:0, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine) were added to the buffer containing RBCs and tolaasin peptides, DDPE facilitated the tolaasin-induced hemolysis while the other two phospholipids showed no effects. At various concentrations of DDPE, the tolaasin-induced hemolysis was stimulated as a dose-dependent manner. The phospholipids composed of mediumchain fatty acids stabilize the tolaasin pore probably by binding between the pore structure and membrane phospholipids and making the membrane thickness thinner around the pore. These results showed that tolaasin molecules make more stable pores in the membrane made with phospholipids composed of medium length fatty acids, suggesting that the length of tolaasin pore is a little shorter than the thickness of RBC membrane.

• Journal of Life Science
• /
• v.28 no.1
• /
• pp.99-104
• /
• 2018

Streptococcus pneumoniae is one of the major pathogens in community-acquired diseases, and it contains several factors that promote its pathogenesis, including pneumolysin (PLY). PLY is a member of the cholesterol-dependent cytolysin family, which attacks cholesterol-containing membranes, thereby forming ring-shaped pores. Thus, it is a major key target for vaccines against pneumococcal disease. We cloned the PLY gene from S. pneumoniae D39 and inserted it into the pQE-30 vector. Recombinant PLY (rPLY) was overexpressed in Escherichia coli M15 and purified by $Ni^{2+}$ affinity chromatography. Similarly, a PLY-EGFP fusion gene was produced by inserting the EGFP gene at the 3' end of the PLY gene in the same vector, and the recombinant protein was purified. Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) showed that both recombinant proteins were purified. rPLY exhibited significant hemolytic activity against 1% human red blood cells (RBCs). Complete hemolysis was obtained at 500 ng/ml, and 50% hemolysis was found with a 240 ng/ml concentration. In contrast, rPLY-EGFP did not show hemolytic activity. However, rPLY-EGFP did bind the RBC membrane, indicating that rPLY-EGFP lost hemolytic activity via EGFP fusion, while retaining its membrane-binding ability. These data suggest that PLY's C terminus is important for its hemolytic activity. Therefore, these two recombinant proteins can be extremely useful for investigating the toxin mechanism of PLY and cell damage during pneumonia.

• Journal of Life Science
• /
• v.25 no.4
• /
• pp.425-432
• /
• 2015

This study was performed to develop high-value-added biomaterials for health and beauty products. Extracts of ethanol and hot water and their subsequent organic solvent fractions were prepared from Lees of Wookukseng (LW), a commercialized Korean traditional rice wine. We investigated their activities on blood coagulation, platelet aggregation, hemolysis against human red blood cells (hRBCs), and anti-oxidation. The water content, pH and brix of the LW were 80.3%, 3.94 and 13.0°, respectively. The yield of ethanol extraction (6.62%) was 3.15 times higher than that of hot-water extraction (2.1%), and the ethyacetate fraction (EAF) of ethanol extract showed the highest content of total polyphenol (128 mg/g) among the various fractions. In anticoagulation activity assay, the EAF of ethanol extract showed a 15-fold extension in TT, PT, and aPTT, indicating that the EAFs contain various inhibitory substances against thrombin, prothrombin and coagulation factors. In anti-platelet aggregation activity assay, the butanol fraction and water residue of ethanol extract showed significant inhibition activity. The activities were comparable to aspirin, a commercial anti-thrombosis agent. The above extracts and fractions did not show hemolysis activity against hRBC up to 5 mg/ml, and had radical scavenging activity against DPPH anion, ABTS cation and nitrite. Our results suggest that the active fractions prepared from LW, which has no specific usage until now, have a high potential as novel resources for anti-thrombosis agents.