• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.205-212
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• 1999

The present study was designed to examine the effects of selenium(Se), vitamin E(Vit. E) and recombinant Bovine Somatotropin(rBST) administration on the selenium and vitamin E concentrations of blood in Hanwoo sires Hanwoo sires were randomly assigned to five groups; 1. control, 2. rBST, 0.09mg/kg body weight (BW) 3. Vit E, 1,500IU/kg BW, 4. Se 0.lmg/kg BW, 5. Vit E, 1.500IU plus Se 0.1mg/kg BW. rBST, Vit. E and Se for each experimental group were given 6 times at 15 days interval by intramuscular injection. Blood samples were collected 10 times for experimental periods, separated the serum by centrifugation, and stored at －7$0^{\circ}C$. Se and Vito E concentrations in blood were measured by fluorophotometer and HPLC. Se concentrations of blood in control, rBST, Vito E, Se and Se plus Vito E groups were 64.55, 65.50, 68.15, 73.11 and 74.09 ppb/$m\ell$, respectively. Se concentration in Vit. E plus Se group was significantly higher than in control and rBST groups (P＜0.05), but Vito E group was not significantly different in control and rBST groups(P＞0.05). The Vit. E concentrations of blood in control, rBST, Vit. E, Se and Se plus Vit. E groups were 2.27, 2.32, 2.80, 2.58 and 2.75 ppm/$m\ell$, respectively. Vit. E and Vit. E plus Se groups were slightly higher than those of any other groups, but not significantly difference in 외 I experimental groups(P＜0.05). These results indicate that Se and Vit. E concentrations of blood were slightly increased with the injection of Se and Vit. E in Hanwoo sires.

• Clinical and Experimental Pediatrics
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• v.46 no.6
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• pp.548-553
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• 2003

Purpose : Matrix metalloproteinase(MMP)-9 is known to breakdown the blood-brain barrier by degrading the extracellular matrix of the subendothelial basement membrane in meningitis. Tissue inhibitor of metalloproteinase(TIMP)-1, a known inhibitor of MMP-9, has been postulated to inhibit the proteolytic activity of MMP-9 by bindng to MMP-9, but their interaction has not been fully understood yet. So far, there have been some reports on the relationship of MMP-9 and TIMP-1 in bacterial meningitis, but few reports in viral meningitis. Furthermore, there has been no report on this in Korea. We investigated the concentrations of MMP-9 and TIMP-1 in cerebrospinal fluid (CSF) and serum of patients with viral meningitis and control subjects, and evaluated their relationship with other clinical parameters of meningitis. Methods : CSF and blood were obtained from 25 subjects with viral meningitis and 14 control subjects. After centrifugation, supernatants were stored at $-20^{\circ}C$ and we assayed concentrations of MMP-9 and TIMP-1 by the sandwich ELISA method. Results : Concentrations of CSF MMP-9 and TIMP-1 were significantly elevated in patients with viral meningitis, when compared with those in control subjects. Their serum levels showed no differences between the two groups. MMP-9 levels were closely correlated with TIMP-1 levels in the CSF($r_s=0.42$, P<0.05). CSF MMP-9/TIMP-1 ratios were significantly higher in patients with viral meningitis than those in the control subjects(P<0.05). Both CSF MMP-9 and TIMP-1 levels positively correlated with CSF total leukocyte counts($r_s=0.43$, P<0.05, $r_s=0.48$, P<0.05). TIMP-1 levels positively correlated with total protein concentrations in the CSF($r_s=0.43$, P<0.05). Conclusion : MMP-9 and TIMP-1 may play an important role in the breakdown and maintenance of BBB in viral meningitis, respectively.

• The Journal of Korean Society for Radiation Therapy
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• v.19 no.2
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• pp.83-90
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• 2007

Purpose: In this study, it was investigated whether commercially produced beer is able to prevent a lymphocyte from radiation induced apoptosis. Materials and Methods: Whole blood samples were acquired from 5 healthy volunteers (male, 26$\sim$38 years old) and the lymphocyte were isolated by density gradient centrifugation. Radiation induced apoptosis of the lymphocyte were investigated by 0.5 Gy, 1.0 Gy, 2.0 Gy, 3.0 Gy to 5.0 Gy irradiation. In some experiments, the donor drunk beer and then blood samples were collected. In other experiments, melatonin or glycine betain was added to lymphocyte culture medium. Treated or untreated lymphocytes were cultured for 60 hours and radiation induced apoptosis of the lymphocyte was analyzed by annexin-V staining through flow cytometery. Results: Relative radiation induced apoptosis ratio of the untreated lymphocytes is 1.22$\pm$1.1, 1.22$\pm$1.1, 1.38$\pm$1.0, 1.47$\pm$1.1, 1.50$\pm$1.2 by radiation dose of 0.5 Gy, 1.0 Gy, 2.0 Gy, 3.0 Gy and 5.0 Gy respectively. Relative radiation induced apoptosis ratio of lymphocytes is isolated from beer drunken donors is 0.97$\pm$1.0, 0.99$\pm$1.0, 1.11$\pm$0.9, 1.29$\pm$1.1, 1.15$\pm$1.1 by radiation doses respectively which are reduced 21.5% compared with untreated lymphocyte. Relative radiation induced apoptosis ratio of the lymphocytes is isolated from non-alcohol beer drunken donors is 1.22$\pm$1.1, 1.17$\pm$1.1, 1.13$\pm$1.3, 1.38$\pm$1.2, 1.32$\pm$1.1 by radiation dose of 0.5 Gy, 1.0 Gy, 2.0 Gy, 3.0 Gy and 5.0 Gy respectively which are reduced 10.8% compared with the untreated lymphocyte. Conclusion: As a result, it is suggested that beer may protect the lymphocyte from radiation damage and inhibit apoptosis.

• Food Science and Preservation
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• v.16 no.3
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• pp.405-411
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• 2009

The manufacturing process and quality properties of Lactococcus lactis-fermented yakchobugak (LFY) containing some colored agro-food products were investigated to develop a good organoleptic bugak from the leaf of the medicinal herb. The de-oiling rate of LFY by centrifugation was 37.6%, which was significantly different to the 3.5% value obtained after standing and the 8.9% value obtained with the beating method (p<0.05). Reducing power (RS) increased with addition of increasing levels of ethanol extracts from agro-food powders. The RS of green tea extract-supplemented LFY had the highest value of 0.97 at $500{\mu}g/mL$, and this was significantly different to values obtained using other extracts (p<0.05). At 83.84%, the DPPH (2,2-diphenyl-1-picrylhydrazyl) free-radical scavenging activity of green tea extract-supplemented LFY was the greatest of all samples tested, followed by LFY with woolgeom at 39.48%, LFY with black rice at 28.45%, and LFY with bokbunja at 22.24% all ethanol extracts were added at $50{\mu}g/mL$ (p<0.05). Acid values of green tea and bokbunja LFYs stored in transparent PE bags at $60^{\circ}C$ for 7 days were 1.82% and 2.03%, respectively. Textural hardness values of LFYs were less than 250.62, except for LFYs with black rice and woolgeom, but these values increased $2{\sim}3$-fold after lactic acid fermentation. Carbohydrate and protein content of LFYs were in the ranges $58.95{\sim}64.63%$ and $7.50{\sim}8.68%$, respectively. Lipid and calorie contents of woolgeom LFY had the lowest values of 22.02% and 490 Kcal, respectively.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.6 no.1
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• pp.81-97
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• 2000

Objectives: This experimental study was carried out to evaluate the effects of aqueous and methanol extracts of Hedyotis diffusa which has long been used for cancer treatment in oriental medicines on the induction of apoptotic cell death in human lymphoid leukemia cell line, HL-60. Methods: Cells were treated with various concentrations (200 to $0.4{\mu}g$) and periods (6 to 30 hr) of $H_2O$ and methanol extracts of Hedyotis diffusa. Then, cells were tested for viability by MTT assay. Cells wrere treated with $200{\mu}g/ml$ of methanol extract fork various periods. Genomic DNA was isolated, separated, on 1.5% agarose gels, stained with ethidium bromide and visualized under UV light. Cells were treated with $200{\mu}g/ml$ of each extract for 16 hr. Then, cells were treated with Hoechst dye 33342 and observed by fluorescence microscopy. Cells were treated with various doses of each for 12 hr and $100{\mu}g/ml$ of methanol extract for various periods. Lysate from the cells used to measure the activity of Caspase-1 and-3 proteases by using fluorogenic peptide substrates including acetyl-YVAD-AMC and acetyl-DEVD-AMC, respectively. Cells were treated with $200{\mu}g/ml$ of each extract for various periods. Cell lysates were immunoprecipated with anti-JNKl antibodies. The immune complex was reacted with $32^p-ATP$ and c-Jun as a substrate. The phosphotransferase activity of JNKI was measured by using PhosphoImage analyzer (Fuji Co., Japan). Nuclear extracts were isolated and incubated with oligonucleotide probe of $NF-{\kappa}B$. Transcriptional activation of ${\kappa}B$ was measured by using EMSA and visualized by PhosphoImage analyzer (Fuji Co, Japan). Cell lysates were prepared and analyzed by Western blotting with anti-Bc12 antibodies and anti-Bax antibodies. Cells were pretreated with various doses of methanol extract for 2 hr. Then, the extract was removed by centrifugation. Cells were resuspended with RPMI-1640 media containing 0.3% agarose, 10% FBS, overlayred onto bottom layer agarose and incubated at $CO_2$ incubator for 6 days. The number of colony was counted under light microscopy ($\time100$). Results: The death of HL-60 cells was markedly induced by the addition of methanol extract of Hedyotis diffusa in a dose and time-dependent manners. The apoptotic characteristic ladder pattern of DNA strand break was observed in death of HL-60 cells. In addition, it was shown nucleus chromatin condensation and fragmentation under Hoechst staining. Therefore, Hedyotis diffusa extract-induced death of HL-60 cells is mediated by apoptotic signaling processes. The activity of Caspase 3-like proteases remained in a basal level in HL-60 cells treated with aqueous extract of Hedyotis diffusa. However, it was markedly increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. In addition, the phosphotransferase activity of JNKl was increased in HL-60 cells treated with methanol extract of Hedyotis diffusa. Furthermore, the activation of transcriptional activator, $NF-{\kappa}B$ was markedly induced by methanol extract of Hedyotis diffusa. Anti-apoptotic Bc12 was cleaved into 23Kda fragment by treatment of methanol extract of Hedyotis diffusa. However, expression of proapoptotic Bax protein was increased by treatment of methanol extract of Hedyotis diffusa in a time-dependent manner. Furthermore, methanol extract markedly inhibited the colony forming efficiency of HL-60 cells in semisolid agar culture. Conclusions: Above results suggest that methanol extract of Hedyotis diffusa induces the apoptotic death of human leukemic HL-60 cells via activations of Caspase-3 proteases, JNKI, transcriptional activator $NF-{\kappa}B$, In addition, our results also suggest that methanol extract of Hedyotis diffusa reduces the malignant potential of HL-60 cells via down regulation of colony forming effciency through cleavage of Bc12 as well as induction of Bax.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.3
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• pp.454-458
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• 2014

In this study, we examined the ionization rate and permeability of nanocalcium prepared from oyster shells with various particle sizes. Four particle sizes of the calcium samples were prepared by centrifugation according to their density disparity in alcoholic solution: NC (normal calcium), C-1 (supernatant of 1,000 rpm), C-2 (supernatant of 2,000 rpm), and C-3 (supernatant of 3,000 rpm). Particle sizes of NC, C-1, C-2, and C-3 were $2,280.3{\pm}64.3nm$, $521.3{\pm}83.3nm$, $313.9{\pm}29.5nm$, and $280.0{\pm}3.4nm$, respectively. C-3 showed a slight increase in ionization rate compared with the other calcium samples, but their differences were not significant. Dialysis membrane-employed analysis showed that nanocalcium permeability increased as its particle size smaller; 32% of C-3 nanocalcium was transported to the outside of the membrane, whereas C-1 showed a 25% transport rate. We determined the permeability of the nanocalciums by using rat intestinal sacs, in order to provide different intestinal environments depending on pH level. Nanocalcium generally showed a higher permeability at pH 7, which represents an ileum environments compared to the duodenum and jejunum environments at pH 4.2 and pH 6.2, respectively. However, C-3 calcium showed the highest permeability, followed by C-2, C-1 and NS calciums. This result shows that the size of calcium positively affected its permeability in the intestinal sac. Taken together, nano-sized calcium derived from discarded oyster shell shows improved permeability in intestinal environments.

• Restorative Dentistry and Endodontics
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• v.27 no.5
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• pp.463-472
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• 2002

Bacterial lipopolysaccharide (LPS) plays a major role in stimulating the synthesis and release of the principal osteoclast-activating cytokines, namely, interleukin 1 and tumor necrosis factor-$\alpha$ from immune cells. Although rnonocytes/macrophages are the main producers of these cytokines, recent evidence has indicated that polymorphonuclear leukocytes (PMN) have the ability to release IL-1 and TNF-$\alpha$. Calcium hydroxide has been shown to be an effective medicament in root canal infections, reducing the microbial titre within the canal. It has been proposed that the therapeutic effect of Ca(OH)$_2$ may also be the result of direct inactivation of LPS. The purpose of this study was to investigate whether treatment of Porphyromonas endodontalis LPS with calcium hydroxide alters its biological action as measured by human PMN secretion of IL-1 and TNF-$\alpha$, and it was compared with Escherichia coli LPS. P. endodontalis ATCC 35406 was cultured in anaerobic condition, and LPS was extracted using the hot-phenol water extraction method and purified. Purchased E. coli LPS was also purified. 100 $\mu\textrm{g}$/ml of each LPS in pyrogen free water were incubated with 25mg/ml Ca(OH)$_2$ at 37$^{\circ}C$ for 7 days. The supernatants were subjected to ultrafiltration, and the isolates were lyophilized and weighed. PMNs were obtained from peripheral blood by centrifugation layered over Lymphoprep. The cells were resuspended (4$\times$10$^6$ cells/ml) in RPMI 1640 followed by treatment with various concentrations of LPS (0, 0.1, 1, 10$\mu\textrm{g}$/ml) for 24 hours at 37$^{\circ}C$ in 5% $CO_2$ incubator. The supernatants of cells were collected and the levels of IL-1$\alpha$, IL=1$\beta$ and TNF-$\alpha$ were measured by enzyme-linked immunosorbent assay. The results were as follows ; 1. The levels of IL-1$\alpha$, IL-1$\beta$, TNF-$\alpha$ from PMN treated with each LPS were significantly higher than those released from unstimulated PMN of the control group (p<0.05). 2. The levels of all three cytokines released from PMN stimulated with each calcium hydroxide treated LPS were significantly lower than those released from PMN stimulated with each untreated LPS (p<0.05), while they were not significantly different from those released from unstimulated PMN of the control group (p>0.05) 3. The levels of secretion for all three cytokines were affected in a dose-dependent manner in PMN stimulated with each LPS (p<0.05), but not in PMN stimulated with each calcium hydroxide treated LPS (p>0.05). 4. The levels of all three cytokines released from PMN stimulated with p. endodontalis LPS were significantly lower than those released from PMN stimulated with E coli LPS (p<0.05).

• The Korean Journal of Pharmacology
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• v.27 no.1
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• pp.33-43
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• 1991

To study the age dependent change of Na, K-ATPase in the erythrocyte of hypertensive rat, 1-kidey 1-clip hypertensive rat was made by the removal of right kidney and partial ligation of left renal artery. After 4 weeks, aged erythrocyte fraction was separated by density gradient centrifugation, and Na, K-ATPase activity and $^3H-ouabain$ binding with ghost cell membrane and ouabain sensitive Rb-uptake with whole cell were measured. 1) In the hypertensive rats, blood pressure was significantly increased to 165.5/119.0 mmHg (systolic/diastolic). Mean corpuscular volume and membrane protein(mg) per $10^9RBC$ were decreased and hemoglobin content was increased in the aged erythrocyte. 2) Na, K-ATPase activity in the solution containing 110 mM NaCl and 10 mM KCI, was decreased in hypertensive rat, and decreased in aged erythrocyte of both group. 3) Ouabain sensitive Rb-uptake by low RbCl concentration(4 mM) was slightly decreased in aged erythrocyte compared to that in young erythrocyte of each group, but slightly increased in young erythrocyte in hypertensive rat compared to that in normotensive rat. 4) Ouabain sensitive Rb-uptake by high RbCl concentration(16 mM) was decreased about 30% to 50 % in aged erythrocyte in both group. And in hypertensive rat, especially in young erythrocyte it was significantly decreased compared to that in normotensive rats. 5) $^3H-ouabain$ binding at 0.13 or $1{\times}10^-6M$ ouabain concentration was slightly decreased in aged erythrocyte of normotensive rat, and significantly decreased in aged erythrocyte of hypertensive rats. 6) $^3H-ouabain$ binding at 6 or $64{\times}10^-6M$ ouabain concentration is slightly decreased in aged erythrocyte of both group, but significantly decreased in young and aged erythrocyte of hypertensive rats compared to that of normotensive rats. The present results suggest that (1) in the young erythrocyte of hypertensive rat, the alterations of Na-pump activity that slightly increased in weak stimulation and inhibited in strong stimulation, may be related to increased molecular activity and the decrease in the number of low affinity site without change in high affinity site, (2) in the aged erythrocyte of normotensive rat, inhibited Na-pump may be related to the change in molecular activity of pump. (3) And in the aged erythrocyte of hypertensive rat, it may be related to the decrease in the number of high and low affinity site as well as the change in molecular activity

• The Korean Journal of Nuclear Medicine Technology
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• v.15 no.1
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• pp.113-116
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• 2011

Purpose: Acetylcholine receptor antibodies cause acetylcholine receptor loss, which is responsible for failure of the neuromuscular junction in the acetylcholine receptor autoantibody. The disease characterized by muscle weakness and fatigue, myasthenia gravis(MG) occurs when the body inappropriately produces antibodies against acetylcholine receptors, and thus inhibits proper acetylcholine signal transmission. And this reason, the measurement of acetylcholine receptor antibodies can be of considerable value in disease diagnosis. Methods: From 2010. August to September, we tested orderd AchRAb 19 samples to get the results. 1. Pipette $5{\mu}{\ell}$ undiluted patient sera and kit control and add 125I AChR $50{\mu}{\ell}$ and incubate at R.T for 2 hours. 2. Pipette $50{\mu}{\ell}$ of anti-human IgG into each tube, and incubate at $2{\sim}8^{\circ}C$ for 2 hours. 3. Pipette $25{\mu}{\ell}$ precipitation enhancer into each tube and add 1mL washing solution into all tubes. 4. Centrifuge each tube for 20minutes at $2{\sim}8^{\circ}C$ at 1500g. 5. Aspirate or decant the supernatant. 6. Pipette 1 mL washing solution into all tubes and resuspend the pellet and repeat centrifugation. 7. Aspirate or decant the supernatant and count all tubes on a gamma counter. Results: Cut off value is 0.2 nmol/L and the results taken below 0.2 nmol/L are negative, the results above that identified as being positive values. We assayed the 19 patients samples and got 7 positive results. Of which, 6 patients were diagnosed as MG.(85.7%). Conclusions: Acetylcholine Receptor autoantibody test is intended for use by persons only for the quantitative determination of it in human serum. Even if measurement of the antibodies is not a routine test, it can be of considerable value in disease diagnosis.

• Korean Journal of Food Science and Technology
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• v.36 no.6
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• pp.893-899
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• 2004

Method for sample preparation and quantitative analysis of 19 permitted and non-permitted synthetic colors in foods was developed based on reversed-phase ion-pairing high performance liquid chromatography. For color extraction of samples, deionized water was added, and pH was appropriately adjusted with 1% ammonia water. Any undissolved matters were extracted with 50% ethanol or 70% methanol. Lipid in snacks was first removed using n-hexane with centrifugation, water was added to extract colors, followed by clean-up and concentration using Sep-Pak $C_{18}$ cartridge. Recovery efficiencies at known concentrations of 19 standard food colors spiked into foods were in 90.3-97.9% range far soft drink, 79.2-101.9% for candy, 84.1-103.4% for jelly, 86.4-100.8% for chewing gum, 83.5-103.4% for ice cream, and 78.5-95.6% for snack.