• The Korean Journal of Mycology
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• v.19 no.1
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• pp.32-40
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• 1991

Mitochondria in Pleurotus ostreatus were isolated and purified by the stepped sucrose density gradient centrifugation, to investigate the effects of the light on the enzymatic activity of the mitochondrial ATP synthase. This enzyme, which was illuminated by the light ranging from 400 nm to 700 nm, showed that the specific activity was stimulated at 490 nm for 15 sec. Effects of organic compounds on the mitochondrial ATP synthase were also investigated at the optimum conditions; The activities of this enzyme were increased to 168 percent by the addition of 2,6-dichlo­rophenol indophenol(DCPIP), 224 percent by phenazine methosulfate(PMS), but inhibited 91 per­cent by oligomycin, 14 percent by 2-heptyl-4-hydroxyquinoline-N-oxide(HQNO) and 75 percent by 2,4-dinitrophenol (DNP), respectively. Effects of metal ions of the mitochondrial ATP synthase were investigated at the optimum conditions. The activities of the enzyme were inhibited 35 percent by $Ca^{2+}$, 14 percent by $Co^{2+}$ and 73 percent by $Mn^{2+}$. For effects of anions, the activities of this enzyme were inhibited 80 percent by $CN^{-}$, 52 percent by $SO_{4}\;^{2-}$, 28 percent by each of $CO_{3}\;^{2-}$­and $NO_{3}\;^{-}$, respectively.

• Journal of Life Science
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• v.17 no.5 s.85
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• pp.658-663
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• 2007

Bacillus thuringiensis is a well-known species of entomophathogenic bacteria that is widely used as a biopesticide against many insect pests. It produces parasporal crystals ($\delta$-endotoxin) and endospores during sporulation. In this report, the $\delta$-endotoxin produced by Bacillus thuringiensis BT-1 and BT-2 were characterized by Scanning Electron Microscope(SEM), Transmission Electron Microscope(TEM), SDS-PACE, and solubilization activity by alkaline solution. BT-1, BT-2 were cultured in the GBY medium, and the $\delta$-endotoxin of them was purified with discontinuous sucrose density gradient centrifugation. Their $\delta$-endotoxin was observed by SEM and TEM. Morphologically, the $\delta$-endotoxin of BT-1 was a square and flat type, whose size was $1.73{\mu}m{\times}0.7{\mu}m$, and the $\delta$-endotoxin of the BT-2 was spherical form whose size was $1.1{\mu}m{\times}0.9{\mu}m$ determined by SEM and TEM. The $\delta$-endotoxin of the BT-1 was composed of 28 kDa and 21 kDa, however, it of the BT-2 was composed of 50 kDa, 35 kDa, and 22 kDa bands determined by SDS-PACE. The purified crystals of BT-1 and BT-2 were dissolved gradually in alkaline solution as time goes by, and it was perfectly dissolved after 3 hours. It is supposed that the $\delta$-endotoxin of crystal was converted to a state of activation in the course of time in the intestines of insect.

• Journal of Sericultural and Entomological Science
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• v.25 no.2
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• pp.1-31
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• 1984

Flacherie, as one of the most prevalent silkworm diseases, causes severe economic damage to sericultural industry and its pathogens have been proved to be flacherie virus (FV) and densonucleosis virus (DNV). Multiplications of the viruses in the larvae of the silkworm, Bombyx mori, were studied by the sucrose density gradient centrifugation and electron microscopy. The quantitative and qualitative changes of nucleic acids and proteins were investigated from the midgut and hemolymph in the silkworm larvae infected separately with FV and DNV. The histopathological changes of epithelial cells of infected midgut also were examined by an electron microscope. 1. Purified fractions of FV or DNV in a sucrose density gradient centrifugation yielded one homogenous and sharp peak without a shoulder, suggesting no heterogenous materials in the preparation. Electron microscopy also revealed that FV and DNV were spherical particles, 27nm and 21nm in diameter, respectively. 2. Silkworm larvae showed a decrease in body weight on the 6th day and in midgut weight on the 3rd day after inoculation with FV or DNV. 3. DNA content was higher in the midgut when infected with FV or DNV, but the hemolymph of the infected larvae showed no difference during first 6 days after inoculation, after which DNA concentration declined rapidly. 4. RNA synthesis of silkworm larvae infected separately with FV and DNV was stimulated in the midgut, but RNA content was reduced in the hemolymph at the early stage of virus multiplication. At the late stage of virus multiplication, however, it was extremely reduced in both midgut and hemolymph. 5. The concentration of protein in the midgut and hemolymph of silkworm larvae infected separately with FV and DNV showed no difference from that of the healthy larvae at the early stage of virus multiplication, but it was significantly reduced at the late stage of virus multiplication. 6. There was no difference in the electrophoretic patterns of RNAs extracted from the midgut of healthy or virus-infected larvae. 7. The electrophoresis of proteins extracted from the midgut infected with FV or DNV, when carried out on the 1st and 5th day after virus inoculation, showed no difference from that of the healthy larvae. But, there was an additional band with medium motility in the proteins on the 8th day after virus inoculation, while a band with low mobility shown in the proteins of healthy larvae disappeared in the infected larvae. However, a band with high mobility in the healthy larvae was separated into two fractions in the infected larvae. 8. The electrophoretic pattern of hemolymph proteins of the silkworm larvae infected separately with FV and DNV was similar to that of the healthy larvae, but the concentration of hemolymph proteins in the infected larvae was lower than that of the healthy larvae at the late stage. 9. Two types of inclusion bodies were shown by the double staining of pyronin-methyl green in the columnar cell of the midgut on the 8th day after FV inoculation. 10. Electron microscopy of the infected midgut revealed that the 'cytoplasmic wall' of the goblet cell thickened on the 5th day after FV inoculation and several types of the cytopathogenic structures, such as virus$.$specific vesicles, virus particles, linear structures, tubular structures, and high electron-dense matrices were observed in the cytoplasm of the goblet cell. The virus particles were also observed in the microvilli and the structures similar to spherical virus particles were observed around the virus-specific vesicles, suggesting the virus assembly in the cytoplasm. 11. Fluorescence micrograph of the infected midgut stained with acridine orange showed that the nucleus, the site of DNV multiplication in the columnar cell, enlarged on the 5th day after virus inoculation. 12. Electron microscopic examination of DNV infected midgut revealed that the nucleolus of the columnar cell was broken into granules and those granules dispersed into apical region of the nucleus on the 5th day after virus inoculation. On the 8th day after inoculation, it was also observed that the nucleus of the columnar cell was full with the high electron-dense virogenic stroma which were similar to virus particles. These facts suggest that the virogenic stroma were the sites of virus assembly in the process of DNV multiplication.

• 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.

• 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