• Korean Journal of Microbiology
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• v.24 no.4
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• pp.341-351
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• 1986

R-plasmid(pSBK203, 2.5Mdal) conferring chloramphenicol resistance was isolated from mutiple antibiotic resistant Staphylococcus aureus D-H-1. Bacillus subtilis BD170 was transformed by this plasmid and restriction enzyme clevage sites of this plasmid were mapped for the cloning of chloramphenicol resistance gene. Taq I partial digested fragment of pSBK203(1.3kb) inserted into Cla I site of pBD9 appears to have both regulatory region for induction and structural gene for chloramphenicol resistance whereas Rsa I fragment (1.3kb, both ends are staggered away 0.1Kb from those of Taq I fragment) inserted into Sca I site of pBR322 showed constitutive expression in E. coli. Hinf I, Taq I, and Bgl II restriction enzyme recognition sites are found in both Rsa I fragment and Taq I fragment. Among these, Bgl II recognition site was associated with chloramphenicol resistance.

• Applied Microscopy
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• v.26 no.4
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• pp.431-446
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• 1996

This experiment was performed to study the ultrastructural changes of the juxtaglomerular cell of mice following subcutaneous injection of heavy metallic agents. Male mice were divided into normal and experimental groups. The mice were subcutaneouly injected with $HgCl_2$ (2mg, 5mg or 10 mg/Kg/BW) or with $K_{2}Cr_{2}O_7$(5 mg, 10 mg or 20 mg/Kg/BW). Mice were sacrificed on 6 hours, 3 days and 14 days after the injection. Kidneys were fixed in the 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by refixation in the 1% osmium tetroxide solution. Dehydrated blocks were embedded in araldite mixture. The sections were cut on a LKB-V ultratome, and ultrathin sections stained with uranyl acetate and lead citrate were observed with JEM 100CX II electron microscope. The results were as follow: 1. Juxtaglomerular cell of the experimental groups showed some alterations, especially in the structures of protein synthesis including dilations and degradations of granular endoplasmic reticula, atrophy of Golgi complex, and numerous free ribosomes in the cytoplasm. 2. Juxtaglomerular cells treated groups showed a number of vacuoles, protogranules and some myelin figures in the cytoplasm, especially in the earlier groups. 3. Juxtaglomerular cells of treated groups, contained a large number of secretory granules showing variable electron densities and pleomorphism in later groups (2 weeks). From the above results, it was concluded that, the mercuric chloride or potassium bichromate induces acute renin release from juxtaglomerular cells of the mice, but many juxtaglomerular cells may secrete prematured secretory granules, or the synthetic system of the cell can not perform normal function.

• Journal of the Korean earth science society
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• v.42 no.5
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• pp.536-555
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• 2021

Sea-surface wind is an important variable in ocean-atmosphere interactions, leading to the changes in ocean surface currents and circulation, mixed layers, and heat flux. With the development of satellite technology, sea-surface winds data retrieved from scatterometer observation data have been used for various purposes. In a complex marine environment such as the Korean Peninsula coast, scatterometer-observed sea-surface wind is an important factor for analyzing ocean and atmospheric phenomena. Therefore, the validation results of wind accuracy can be used for diverse applications. In this study, the sea-surface winds derived from ASCAT (Advanced SCATterometer) mounted on MetOp-A/B (METeorological Operational Satellite-A/B) were validated compared to in-situ wind measurements at 16 marine buoy stations around the Korean Peninsula from January to December 2020. The buoy winds measured at a height of 4-5 m from the sea surface were converted to 10-m neutral winds using the LKB (Liu-Katsaros-Businger) model. The matchup procedure produced 5,544 and 10,051 collocation points for MetOp-A and MetOp-B, respectively. The root mean square errors (RMSE) were 1.36 and 1.28 m s^-1, and bias errors amounted to 0.44 and 0.65 m s^-1 for MetOp-A and MetOp-B, respectively. The wind directions of both scatterometers exhibited negative biases of -8.03° and -6.97° and RMSE values of 32.46° and 36.06° for MetOp-A and MetOp-B, respectively. These errors were likely associated with the stratification and dynamics of the marine-atmospheric boundary layer. In the seas around the Korean Peninsula, the sea-surface winds of the ASCAT tended to be more overestimated than the in-situ wind speeds, particularly at weak wind speeds. In addition, the closer the distance from the coast, the more the amplification of error. The present results could contribute to the development of a prediction model as improved input data and the understanding of air-sea interaction and impact of typhoons in the coastal regions around the Korean Peninsula.

• Herbal Formula Science
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• v.24 no.3
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• pp.163-174
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• 2016

Objectives : Rheum undulatum Linne and Glycyrriza uralensis Fischer are widely used herbal medicine. In this study, anti-oxidant and liver protective effects of R. undunlatum extract (RUE) and G. uralensis extract (GUE) were investigated in HepG2 cells, respectively. Oxidative stress and liver fibrosis were induced by arachidonic acid (AA) and iron, and CCl₄.Methods : MTT assay was assessed for cell viability, and immunoblotting analysis was performed to detect expression of apoptosis related proteins. In addition, reactive oxygen species (ROS) and mitochondrial dysfunction were measured. In vivo, BALB/c mouse were orally administrated with the aqueous extract of 10 mg/kg RUE and 100 mg/kg GUE for 3 days and then, injected with CCl₄ 0.5 ml/kg body weight to induce acute liver damage. Serum ALT level was measured, and histological change was observed in Harris's hematoxylin and eosin stainResults : RUE and GUE pre-treatment increased relative cell viability in concentration dependent manner and altered the expression levels of apoptosis-related proteins such as procaspase 3, PARP and Bcl-xL. RUE and GUE also inhibited the mitochondrial dysfunction and excessive reactive oxygen species (ROS) production induced by AA and iron. In addition, RUE and GUE activated liver kinase B1 (LKB1), by increasing phosphorylation. Moreover, RUE and GUE treatment decreased liver injuries induced by CCl₄, as evidenced by decreases in histological liver damage as well as serum alanine amino transferase (ALT) level.Conclusions : These data suggest that RUE and GUE has anti-oxidant and liver protective effects against AA and iron-induced oxidative stress and CCl₄-induced liver injury.

• Journal of the Korean Society of Food Science and Nutrition
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• v.20 no.5
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• pp.455-460
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• 1991

In order to identify hard distinct 12 fish species(shiba shrimp Metapenaeus joyneri, fleshy shrimp Penaeus orientalis, ridgetail prawn Palaemon carinicauda, yellow croaker Pseudosciaena manchurica, croaker Niber albiflora, Colichthyes fragilis, brown sole Limanda herzensteini, frog flounder Pleuronichthys cornutrs, Areliscus rhomaleus, stone flounder Kareius bicoloratus, harvest fish Pampus argenteus, flag fish Goniistius zonatus) by seeing with naked eye in Kunsan coastal area, sarcoplasmic protein in the supernatant was used for isoelectric focusing. For getting supernatant, fish muscle tissue was blended with two times deionized water and centrifuged (at $4^{\circ}C$, 12,000rpm for 15min). Isoelectric focusing of sarcoploasmic protein carried out on a LKB Multiphor II using polyacrylamide gel plate (2mm thickness, pH $3.5~10^{\circ}C$, pH 5~8 gradient, at $10^{\circ}C$ for 1.5, 3 hours). In case of uncertain protein pattern, pH gradient was modified to narrow pH gradiet, and excuted 2-D electrophoresis using conventional polyacrylamide gel electrophoresis. Most of fishes except yellow croaker and Collichthyes fragilis were distingushed by isoelectric focusing. The protein maps of 2-D electrophoresis for analyzing two protein bands at aimilar positions(pH 5, 6) between the two fish species showed the diffeences of the estimated molecular weights, 11,700(pH5.0) and 87,000(pH6.0)

• Applied Microscopy
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• v.24 no.2
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• pp.63-77
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• 1994

This experiment was performed to study the morphological responses of the epidermis of the rat scalp, following X-ray irradiation. Male rats were divided into normal and experimental groups. Rats anesthetized with sodium thiopental, were exposed only on their head areas with a single dose of 3,000rads or 6,000rads, respectively. Radiation was produced by Mitsubishi Linea Accelerator ML-4MV at the speed of 200rads/min. The target distance was 80cm. Animals were sacrificed on six hours, two days and six days following irradiation. By the perfusion fixation through the heart, rats were fixed with 1% glutaraldehyde-1% paraformaldehyde solution. Pieces of the tissue taken from the scalp were refixed in 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide, and embedded within araldite mixture. The sections were cut on a LKB-V ultratome, stained with uranyl acetate and lead citrate, and were observed with JEM 100CX-II electron microscope. The results were as follow; 1. Six hours after exposure to 3,000rads of X-ray. Disrupted intercellular spaces, within which some amorphous materials were filled, disrupted mitochondria, and vacuoles in the keratinocytes were frequently observed, but six days after exposure to 3,000rads of X-ray, Morphology of the keratinocytes was generally restored. 2. Many of the morphological changes were seen on the six days after exposure to 6,000rads of X-ray. 3. Widened intercellular spaces and thickened dense plaques of the desmosomes were frequently observed after exposure to 6,000rads of X-ray. 4. In the experimental groups, the Langerhans and the Merkel cells were damaged, similarly to the keratinocyte. Above results suggest that head irradiation with the dose of 3,000rads temporarily damaged the epidermis of the scalp, though most of the structures recover within six days, whereas with the dose of 6,000rads it severely damaged the epidermis without showing any recovering tendency.

• Applied Microscopy
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• v.20 no.1
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• pp.36-50
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• 1990

This experiment was performed to study the morphological responses of the enterochromaffin cells in the duodenal mucosa of rabbit following bile duct ligation. Adult male rabbits were divided into normal, sham operation and experimental groups. Bile duct ligation was performed under ether anesthesia and animals were sacrificed on the 1st, 3rd, 5th, 7th and 14th day after operation. Mucosal specimens from the duodenum were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde(0.1M Millonig's phosphate buffer, pH 7.3), followed by postfixation with 1% osmium tetroxide(0.1M Millonig's phosphate buffer, pH 7.3), and embedded within araldite mixture. The sections were cut on a LKB-V ultratome, and observed under a JEM 100CX II electron miroscope. The results were as follow; 1. Irregularities of the nuclei of the enterochromaffin cells were noticed from the 1st day after bile duct ligation, and concentration of the chromatin in the nucleus was more frequently observed on the 7th and the 14th day. 2. The granular endoplasmic reticulum and Golgi complex of the enterochromaffin cell were more developed than those of the normal on the 1st day after bile duct ligation, but they showed poor organization from the 3rd day. 3. Amount of the microfilaments in the enterochromaffin cell was significantly increased from the 5th day after bile duct ligation and they were more frequently observed in the vicinity of the nucleus. 4. Vacuoles with various electron densities in the enterochromaffin cell were increased in number from the 3rd day after bile duct ligation. 5. Glycogen-like particles in the enterochromaffin cell were frequently observed on the 3rd and the 5th day after bile duct ligation. 6. In the early stage of the ligation of bile duct, in the enterochromaffin cells, well developed intracellular organelles, such as granular endoplasmic reticulum and Golgi apparatus were pronounced. But, in the later stage, the cells contained poor organelles, with the some structural sign of necrotic change.

• Herbal Formula Science
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• v.29 no.2
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• pp.71-80
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• 2021

Objectives : This study investigated cellular-protective effects of Nardotidis seu Sulculii Concha water extract (NSCE) against oxidative stress induced by arachidonic acid (AA)+iron or tert-butylhydroperoxide (tBHP). Methods : In vitro, MTT assay was assessed for cell viability, and immunoblotting analysis was performed to detect expression of AMP-activated kinase (AMPK) signaling pathway and autophagy related proteins. In vivo, mice were orally administrated with the aqueous extract of NSCE of 500 mg/kg for 3 days, and then injected with CCl₄ 0.5 mg/kg body weight to induce acute damage. The level of liver damage was measured by serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) analysis. Results : Treatment with NSCE inhibited cell death induced by AA+iron and tBHP. NSCE induced the phosphorylation of AMPK, and this compound also induced the phosphorylation of LKB1, an upstream kinase of AMPK, and Acetyl-CoA carboxylase (ACC), a primary downstream target of AMPK. NSCE increased the protein levels of autophagic markers (LC3II and beclin-1) and decreased the phosphorylation of mammalian target of rapamycin (mTOR) and simultaneously increased the phosphorylation of unc-51-like kinase-1 (ULK-1) in time-dependent manner. Conclusions : NSCE has the ability 1) to protect cells against oxidative stress induced by AA+iron or tBHP. NSCE 2) to activate AMP-activated protein kinase (AMPK), and 3) to regulate autophagy, an important regulator in cell survival.

• Journal of Korean Medicine Rehabilitation
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• v.32 no.2
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• pp.19-36
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• 2022

Objectives This study is to investigate the effects and mechanisms of Imyo-san (IMS) on the obese mice model induced by high-fat diet. Methods Antioxidative capacity was measured by in vitro method. C57BL/6 mice were randomly assigned into 5 groups (n=7). Normal group was fed general diet (Normal). The other 4 groups were fed high fat diet (HFD) with water (Control), with Garcinia gummi-gutta (GG, Garcinia gummi-gutta 200 mg/kg), with low-dose IMS (IMSL, Imyo-san 0.54 g/kg) and with high-dose IMS (IMSH, Imyo-san 1.08 g/kg). Results IMS showed high radical scavenging activity. After 6 week experiment, body weight, food intake, food efficiency ratio (FER), epididymal fat and liver weight, triglyceride (TG), total cholesterol (TC), high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol, very low density lipoprotein (VLDL) cholesterol, sterol regulatory element-binding protein-1 (SREBP-1), phospho-acetyl-CoA carboxylase (p-ACC), fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1), SREBP-2, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), phospho-liver kinase B1 (p-LKB1), phospho-AMP-activated protein kinase (p-AMPK), peroxisome proliferator-activated receptor 𝛼 (PPAR𝛼), peroxisome proliferator-activated receptor 𝛾 coactivator-1𝛼 (PGC-1𝛼), uncoupling protein-2 (UCP-2), carnitine palmitoyltransferase 1A (CPT-1A), and histology of liver and epididymal fat were measured and analysed. Body weight gain, FER, liver and epididymal fat weight of IMS groups were significantly decreased. There were significant improvements in blood lipids with less TG, TC, LDL-cholesterol, VLDL-cholesterol and more HDL-cholesterol. Proteins associated with lipid synthesis (SREBP-1, p-ACC, FAS, SCD-1) and cholesterol (SREBP-2, HMGCR) was improved. Factors regulating lipid synthesis and lipid catabolism (p-LKBI, p-AMPK, PPARα, PGC-1α, UCP-2, CPT-1A) were increased. In histological examinations, IMS group had smaller fat droplets than control group. All results increased depending on concentration. Conclusions It can be suggested that IMS has anti-obesity effects with improving lipid metabolism.

• Applied Microscopy
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• v.24 no.2
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• pp.78-92
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• 1994

Lysozyme has been reported to be present in the secretory granules of the Paneth cell, and lysozyme immunoreactivity has been detected by immunogold method in Paneth cells of the intestine of human, mouse and rat. The present study was aimed at clarifying the intracellular distribution and changes of the lysozyme immunoreactivity in rabbit Paneth cell after common bile duct ligation of rabbit, using the electron microscope immunogold technique. Healthy adult rabbits weighing about 2kg body weight were divided into normal and bile duct ligated groups. Common bile duct ligation was performed aseptically under ether anesthesia. Experimental animals were sacrificed on the 1st, the 3rd, the 5th, the 7th and the 14th day after the operation. Mucosal specimens from the intestinal gland of ileum were fixed in 2.5% glutaraldehyde-1.5% paraformaldehyde, followed by 1% osmium tetroxide, embedded in araldite mixture, cut with LKB-V ultratome. Ultrathin sections were placed on parlodion coated nickel grids (200mesh). The section-bearing grids were floated upside down on the added substance in a moist chamber at room temperature except for the primary antibody step, which was at $4^{\circ}C$. Sections were etched with a saturated solution of sodium m-periodate for 60min. After etching, sections were pretreated with 0.02M tris buffered saline (TBS), pH 8.4, with 1% bovine serum albumin (BSA, Sigma) for 60min, then treated polyclonal rabbit anti-human lysozyme (Dakipatts) diluted 1 : 50 in TBS with 0.1% BSA for 20hr. Subsequently, grids were incubated 60min in biotinylated goat anti rabbit IgG (Amersham) diluted 1 : 100 in TBS with 0.1% BSA. After this, sections were incubated 60min on streptavidin gold G10 (Amersham) diluted 1 : 50 in TBS with 0.1% BSA. After each step, the grids were briefly rinsed with TBS with 0.1% BSA. After the strepavidin gold step, the sections were jet washed with distilled water. Counterstain of the sections performed by uranyl acetate and lead citrate, and observed with JEM 100 CX II electron microscope. Observed results were as follow; 1. Secretory granules of mouse Paneth cells have a lysozyme immunoreactivity and also eosinophil leucocyte of rabbit applied for the positive-control stain, are well labeld with gold particles. 2. Normal rabbit Paneth cells have a lysozyme immunoreactivity restricted on the secretory granules. 3. Amount lysosomes containing myelin figures in the Paneth cells were significantly increased from 5th day after the common bile duct ligation. 4. Immunoreactivity of Paneth cell secretory granules were more activated on the 3rd day after the common bile duct ligation as compared with those of the normal animal. But the lysozyme immunoreactivity were decreased from the 5th day after the common bile duct ligation. 5. Considering the above finding, lysozyme contained Paneth cell are affected following of common bile duct ligation, whereas lysosomes containing myelin-figure do not exhibit any immunoreactive relationship with those of secretory granules.