• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.245-252
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• 2002

This study was to evaluate the in vitro survival of bovine enucleated MII (eMII) oocytes according to minimum volume cooling (MVC) freezing method and activation timing, and their in vitro development after somatic cell nuclear transfer (SONT). in vitro matured bovine oocytes for 20 h were stained with 5 $\mu\textrm{g}$/$m\ell$ Hoechst, and their 1st polar body and MII plate were removed by enucleation micropipette under UV filter. Also, eMII oocytes were subjected to activation after (group II) and before (group III) vitrification in 5 ${\mu}{\textrm}{m}$ ionomycin added CRlaa medium for 5 min. For vitrification, eMll oocytes were pretreated with EG10 for 5 min, exposed to EG30 for 30 sec and then directly plunged into L$N_2$. Thawing was taken by 4-step procedures at 37$^{\circ}C$. Survived eMII oocytes were subjected to SONT with cultured adult bovine ear cells. Reconstructed oocytes were cultured in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide and 2.5 $\mu\textrm{g}$/$m\ell$ of cytochalasin D added CRlaa medium for 1 h, and then in 10 $\mu\textrm{g}$/$m\ell$ of cycloheximide added CRlaa medium for 4 h. Subsequently, the reconstructed oocytes were incubated for 2 days and cleaved embryos were further cultured on cumulus-cell monolayer drop in CRlaa medium for 6 days. Survival rates of bovine vitrified-thawed eMII oocytes in group II (activation after vitrification and thawing) and III (activation before vitrification) were 81.0% and 84.9%, respectively. Fusion rates of cytoplasts and oocytes in group II and III were 69.0% and 70.0%, respectively, and their results were not different with non-frozen NT group (control, 75.2%). Although their cleaved rates (53.4% and 58.4%) were not different, cytoplasmic fragment rate in group II (32.8%) was significantly higher than that in group III (15.6%)(P<0.05). Also, subsequent development rate into >morula in group II (8.6%) was low than that in group III(15.6%). However, in vitro development rate in group III was not different with that in control (24.8%). This result suggested that MVC method was appropriate freezing method for the bovine eMII oocytes and vitrified eMII oocytes after pre-activation could support in vitro embryonic development after SONT as equally well as fresh oocytes.

• The Korean Journal of Physiology
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• v.29 no.1
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• pp.69-80
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• 1995

Renal proximal tubular hypertrophy and hyperfunction are known to be early manifestations of experimental and human diabetes. As the hypertrophy and hyperfunction have been suggested to be central components in the progression to renal failure, an understanding of their underlying causes is potentially important for the development of therapy. A primary rabbit kidney proximal tubule cell culture system was utilized to evaluate the possibility that the renal proximal tubular hypertrophy and hyperfunction observed in vivo in diabetes mellitus, can be attributed to effects of elevated glucose levels on membrane transport systems. Primary cultures of rabbit proximal tubules, which achieved confluence at 10 days, exhibited brush-border characteristics typical of proximal tubular cells. Northern analysis indicated $2.2{\sim}2.3$ and 2.0 kb Na/glucose cotransporter RNA species appeared in fresh and cultured proximal tubule cells after confluence, repectively. The cultured cells showed reduced Na/glucose cotransporter activity compared to fresh proximal tubules. Primary cultured proximal tubule cells incubated in medium containing 20 mM glucose have reduced ${\alpha}-MG$ transport compared to cells grown in 5 mM glucose. In the proximal tubule cultures incubated in medium containing 5 mM or 20 mM glucose, phlorizin at 0.5 mM inhibited 0.5 mM ${\alpha}-MG$ uptake by 84.35% or 91.85%, respectively. The uptake of 0.5 mM ${\alpha}-MG$ was similarly inhibited by 0.1 mM ouabain (41.97% or 48.03% inhibition was observed, respectively). In addition, ${\alpha}-MG$ uptake was inhibited to a greater extent when $Na^{+}$ was omitted from the uptake buffer (81.86% or 86.73% inhibition was observed, respectively). In cell homogenates derived from the primary cells grown in 5 mM glucose medium, the specific activity of the Na/K-ATPase $(6.17{\pm}1.27\;{\mu}mole\;Pi/mg\;protein/hr)$ was 1.56 fold lower than the values in cell homogenates treated with 360 mg/dl D-glucose, 20 mM $(9.67{\pm}1.22\;{\mu}mole\;Pi/mg\;protein/hr)$. Total $Rb^{+}$ uptake occurred at a significantly higher rate (1.60 fold increase) in primary cultured rabbit kidney proximal tubule cell monolayers incubated in 20 mM glucose medium $(10.48{\pm}2.45\;nM/mg\;protein/min)$ as compared with parallel cultures in 5 mM glucose medium. $Rb^{+}$ uptake rate in 5 mM glucose medium was reduced by 28% when the cultures were incubated with 1 mM ouabain. The increase of the $Rb^{+}$ uptake by rabbit kidney proximal tubule cells in 20 mM glucose could be attributed primarily to an increase in the rate of ouabain-sensitive $Rb^{+}$ uptake $(5\;mM\;to\;20\;mM;\;4.68{\pm}0.85\;to\;8.38{\pm}1.37\;nM/mg\;protein/min)$. In conclusion, the activity of the renal proximal tubular Na,K-ATPase is elevated in high glucose concentration. In contrast, the activity of the Nafglucose cotransport system is inhibited.

• Korean Journal of Microbiology
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• v.37 no.3
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• pp.197-203
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• 2001

An ubiquitous bacterium, Pseudomonar cepacia KH410 was isolated from fresh water plant root and identified. Adsorption of heavy metals of lead, cadmium and copper by this strain was investigated. Optimal conditions foradsorption was 1.0 dry g-biomass, at pH 4.0 and temperature of $40^{\circ}C$. Adsorption equilibrium reached max-imum after 120 min in 1000 mg/l metal solutions. The adsorption capacity (K) of lead was 5.6 times higher thancadmium and 4.0 times higher than that of copper. Adsorption of lead was applicable for Langmuir modelwhereas Freundlich model for cadmium and copper, respectively. Adsorption strength (1/n) of heavy metal ionswere in the order of lead>copper>cadmium. Uptake capacity of lead, cadmium and copper by dried cell was83.2,42.0,65.2 mg/g-biomass, respectively. Effective desorption was induced 0.1 M HCI for lead and 0.1 $HNO_3$ for cadmium and copper. Pretreatment to increase ion strength was the most effective with 0.1 M KOH.Uptake by immobilized cell was 77.8,58.5,71.2 mg/g-biomass for lead, cadmium and copper, respectively. Theimmobilized cell was more effective than ion exchange resin on removal of heavy metals in solution containinglight metals.

• Korean Journal of Microbiology
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• v.49 no.4
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• pp.309-312
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• 2013

Fungal cell walls consist of various glucans and chitin. Fungi produce chitinases for their growth and development. The inky cap, Coprinellus congregatus, produces at least two different chitinases during its life cycle. Chitinase 1 (chi1) is expresses throughout its life cycle while chitinase 2 (chi2) is expressed at the mushroom autolysing phase. The cloned cDNA of chi1 is successfully expressed as a fusion protein with c-myc in Pichia pastoris, and purified by the affinity chromatography. The optimum pH and temperature of Chi1 was pH 8.0 and $35^{\circ}C$, respectively when 4-nitrophenyl N,N',N"-triacetyl-${\beta}$-D-chitotrioside was used as the substrate. The $K_m$ value and $V_{max}$ for the substrate was 0.780 mM and 0.10 OD $min^{-1}unit^{-1}$, respectively. The addition of purified Chi1 resulted in total growth inhibition against several plant pathogenic fungi such as Alternaria alternata, Fusarium graminearum and Trichoderma harzianum at the concentration of 60 ${\mu}g/ml$.

• Membrane and Water Treatment
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• v.6 no.5
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• pp.423-437
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• 2015

Polyvinylidene fluoride/fullerene nanoparticle (PVDF/$C_{60}$) composite microfiltration (MF) membranes were fabricated by a non-solvent induced phase separation (NIPS) using N, N-dimethylacetamide (DMAc) as solvent and deionized water (DI) as coagulation solution. Polyvinylpyrrolidone (PVP) was added to the casting solution to form membrane pores. $C_{60}$ was added in increments of 0.2% from 0.0% to 1.0% to produce six different membrane types: one pristine PVDF membrane type with no $C_{60}$ added as control, and five composite membrane types with varying $C_{60}$ concentrations of 0.2, 0.4, 0.6, 0.8 and 1.0%, respectively. The mechanical strength, morphology, pore size and distribution, hydrophilicity, surface property, permeation performance, and fouling resistance of the six membranes types were characterized using respective analytical methods. The results indicate that membranes containing $C_{60}$ have higher surface porosity and pore density than the pristine membrane. The presence of numerous pores on the membrane caused weaker mechanical strength, but the water flux of the composite membranes increased in spite of their smaller size. Initial flux and surface roughness reached the maximum point among the composite membranes when the $C_{60}$ concentration was 0.6 wt.%.

• Proceedings of the Korean Society of Food Science and Nutrition Conference
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• 2004.11a
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• pp.53-60
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• 2004

The pro-apoptotic effect of phenethyl isothiocyanate (PEITC) and the role of glutathione (GSH) in sulforaphane (SFN)-induced antioxidant response element-dependent gene expression were investigated. The caspase-3 and caspase-9 activities were stimulated by PEITC. The release of cytochrome c was time- and dose- dependent. SP600125 suppressed apoptosis induced by PEITC. Similarly, this JNK inhibitor attenuated both cytochrome c release and caspase-3 activation induced by PEITC. SFN is converted to the glutathione conjugate by glutathione S-transferases (GSTs). It was accumulated in mammalian cells by up to several hundred-fold over the extracellular concentration, by conjugation with intracellular GSH. The induction of ARE by SFN was 8.6-fold higher than by SFN-NAC. The decrease in ARE expression at higher concentrations of SFN and SFN-NAC was correlated with the accelerated apoptotic cell death, with a dose-dependent activation of caspase 3 activity by SFN. Upon addition of extracellular GSH within 6 hr of treatment with SFN, the effect on ARE expression was blocked almost completely.

• Journal of Convergence Korean Medicine
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• v.5 no.2
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• pp.11-16
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• 2023

Objectives: The purpose of this study was to investigate the anti-obesity effects of the aqueous extract of Schizandra chinensis (SC) in menopausal mice. Methods: To induce menopausal obesity, female mice were ovariectomized (OVX) and fed a high-fat diet (HFD; 60% fat, 28% carbohydrates, 14% protein) for 12 weeks. The mice were divided into 6 groups (n = 8): NOR (sham-operated and vehicle-treated), HFD+OVX (vehicle-treated), E2 (17-beta estradiol 50 ㎍/kg-treated), SC1 (1 mg/kg SC-treated), SC10 (10 mg/kg SC-treated), and SC100 (100 mg/kg SC-treated). Samples were orally administered for 6 weeks, after which all experimental mice were sacrificed. Body weight, feeding efficiency, white adipose tissue weight, adipocyte diameter, and fat vacuoles in liver were analyzed. Results: By treating with SC extract, the body weight and feeding efficiency of mice were significantly decreased. The weight of visceral fat tissues was decreased in the SC10 and SC100 groups. Histopathology showed that fat cell diameters of white adipose tissue were also decreased in the SC10 and SC100 groups. Additionally, SC extract regenerated the hepatocyte damage and decreased the size and number of follicular adipocytes Conclusion: In summary, these results suggest that SC has inhibitory effects against menopausal obesity. Schizandra chinensis may be a potential alternative for obesity among female menopausal diseases.

• Korean Chemical Engineering Research
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• v.52 no.1
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• pp.88-91
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• 2014

In this study, oil as a source of biodiesel from Nannochloropsis oceanica was extracted using organic solvent. The oil extraction yield and efficiency from dry and wet microalgae were investigated. The initial fatty acids content of the N. oceanica was 317.8 mg/g cell showing a high oil content over 30%. The yield from dry microalgae was higher than that from wet microalgae due to the inhibition of water. The yield by chloroform-methanol was the highest and the yield by hexane was the lowest. However, the total fatty acids contents with the chloroform-methanol were 678.7 and 778.2 mg/g oil under dry and wet conditions, respectively. The high oil extraction yield by chloroform-methanol reflected the fact that the extracted oil contained a high level of impurity. The hexane-methanol extraction from dry N. oceanica showed high oil extraction efficiency, 82.6%. The chloroform-methanol extraction under wet condition also showed high efficiency, 88.0%. While the hexane-methanol extraction from dry microalgae is desirable under low drying cost, the chloroform-methanol extraction from wet microalgae is desirable under high drying cost.

• Korean Journal of Veterinary Pathology
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• v.2 no.2
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• pp.95-106
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• 1998

The prevalence of primary liver carcinoma (PLC) is relatively high in Clonorchis sinensis (CS) endemic areas in Korea. PLC is a malignant tumor which can be subclassified into hepatocellular carcinoma and cholangiocarcinoma(CC). CC has been associated with clonorchiasis, but it is unclear whether clonorchiasis is associated with hepatocarcinogenesis. This experiment was designed to investigate relationships between clonorchiasis and early changes of hepatocarcinogenesis. Sixteen Sprague-Dawley rats weighing 150g were divided into two groups of 8 rats in each. All rats were fed choline-devoid(CD) diet for 4 weeks. Group 1 was given 0.015-0.020% diethylnitrosamine(DEN) as drinking water for 1 week. After one week, the rats were treated orally with 1% N-acetylaminofluorene(AAF) (5 times per week for 2 weeks). Group 2 was treated equally to group 1 except for CS infection during AAF treatment. Two rats in each group were sacrificed at 4th, 5th, 6th and 7th week of the experiment. Livers were stained with OV -6, proliferating cell nuclear antigen(PCNA) and GST-p. Results were as follows: Group 2 livers showed more oval cell proliferation in parenchyma and portal areas at the 4th, 5th, 6th and 7th weeks than did livers of group 1 (p＜0.01). PCNA was mostly localized in oval cell populations, rather than hepatocytes and biliary cells. The ratio of oval cells to hepatocytes was much higher in group 2 than in group l(p＜0.01 The ratio of hepatocytes to biliary cells is higher in group 2 than in group 1 (p＜0.05), More group 2 acidophilic foci reacted to GST-p monoclonal antibody than in the noninfected group. It appeared that CS infection promoted potentially precancerous acidophilic foci and oval cell proliferation.

• Journal of Korean Neurosurgical Society
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• v.29 no.10
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• pp.1289-1295
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• 2000

Purpose : Recent evidence suggests a possible role for leukocytes in brain injury following ischemia and reperfusion. This study examined the temporal profile of ischemic tissue damage and leukocyte response after transient middle cerebral artery occlusion(MCAO) with reperfusion in the mouse. Methods : Focal cerebral ischemia was made by temporary occluding of the stem of the proximal MCA. Two groups of the mouse were investigated : (1) sham operation(n＝10), and (2)those having the arterial occlusion released after 90 minute(n＝20). By 4 hours(n＝10) and 24 hours(n＝10) after the onset of ischemia-reperfusion, fluorescein videoimages were under-taken in the pial venules of the mouse using a closed cranial window technique. Rhodamine 6G was administered as a $80-100{\mu}l/min$ i.v. loading dose and a $30-40{\mu}l/min$ i.v. maintenance dose in saline to selectively label circulating leukocytes. Neuropathologic evaluation for brain injury was accomplished using the histochemical stain 2,3,5-triphen-yltetrazolium chloride(TTC) and hematoxylin and eosin(H & E) stain. Results : The mean number of adherent leukocytes to cerebral venules in the 90 minutes MCAO and 24 hours reperfusion group were $306{\pm}24$ compared with $72{\pm}8$ in the sham operation group. In the TTC staining method, the cortical infarct affecting 34.8% of hemispheric volume were created in all of animals (n＝10) undergoing 90 minute MCAO with 24 hours reperfusion, but the infarcted area were not found in the other(sham operation and 90 minute MCAO with 4 hours reperfusion)groups. In the H & E stain, the brain tissue following 90 minute MCAO with 4 hours reperfusion revealed only a pyknosis of the nuclei with shrunken cytoplasm, but infiltrated leukocytes were not observed. After 24 hours of reperfusion, a many leukocytes were infiltrated within parenchyma and blood vessles. Conclusions : These findings demonstrate the feasiblity of continous in vivo monitoring of leukocyte adherence in cerebral venules and suggest that reperfusion induced leukocyte adherence to venular endothelium may contribute to tissue injury following focal cerebral ischemia.