• Journal of the Korean Applied Science and Technology
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• v.32 no.4
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• pp.702-711
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• 2015

This study was conducted to determine the feasibility of using Gastrodia elata Blume as a cosmetic raw material by investigating the physiological activities of its extracts, varying the concentration, solvent, and fermentation method (non-fermentation and fermentation using lactic acid bacteria and effective microorganisms). Of the extracts in three different solvents-water, EtOH, and 70% EtOH-at four different concentrations (0.725, 1.25, 2.5, and 5 mg/mL), the EtOH extracts demonstrated the highest contents of antioxidants (flavonoids, polyphenols, and DPPH free radical scavengers). The DPPH free radical scavenging activity in the EtOH extracts of EM-fermented Gastrodia elata Blume increased from $27.08{\pm}0.5%$ at 1.25 mg/mL to $35.89{\pm}0.8%$ at 2.5 mg/mL. The tyrosinase inhibitory activity test was performed to measure skin-whitening capacity and revealed the LB-fermented EtOH extracts to be the most efficacious ($39.1{\pm}0.4%$ at 0.725 mg/mL, $62.8{\pm}1.5%$ at 2.5 mg/mL). Viability was found to exceed 85% in RAW 264.7 cells treated with all extracts (water, EtOH, 70% EtOH at 10, 25, $50{\mu}L$, fermented and non-fermented), thus proving that Gastrodia elata Blume extracts do not cause inflammation. When RAW 264.7 cells were stimulated with lipopolysaccharide as positive controls under the same conditions to determine the antioxidant activity in the presence of reactive oxygen species (ROS), EM-fermentation was found to impart excellent antioxidant capacity. This study verified the physiological activities of fermented Gastrodia elata Blume extracts that are best suited for cosmetic ingredients, such as antioxidants, tyrosinase inhibitors and anti-inflammatory agents.

• Journal of the Korea Organic Resources Recycling Association
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• v.11 no.2
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• pp.55-65
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• 2003

The combined effect of bioaugmentation of dechlorinating bacterial cultures and addition of iron powder($Fe^0$ on reductive dechlorination of tetrachloroethylene(PCE) and other chlorinated ethylenes in a artificially contaminated soil slurry(60micromoles PCE/kg soil). Two different anaerobic bacterial cultures, a pure bacterial culture of Desulfitobacterium sp. strain Y-51 capable of dechlorinating PCE to cis-1,2-dechloroethylene(cis-DCE) and the other enrichment culture PE-1 capable of dechlorinating PCE completely to ethylene, were used for the bioaugmentation test. Both treatments introduced with the strain Y-51 and PE-1 culture (3mg dry cell weight/kg soil) showed conversion of PCE to cis-DCE within 40days. The treatments added with $Fe^0$(0.1-1.0%) alone to the soil slurry resulted in extended PCE dechlorination to ethylene and ethane and the dechlorination rate depended on the amount of $Fe^0$ added. The combined use of the bacterial cultures with $Fe^0$(0.1-1.0%)) showed the higher PCE dechlorination rate than the separated application and the pattern of PCE dechlorination and end-product formation was different from those of the separated application. When 0.1% of $Fe^0$ was added with the cultures, the treatments with the strain Y-51 and $Fe^0$ resulted in cis-DCE accumulation from PCE dechlorination, but the treatment with the enrichment culture and $Fe^0$ showed the more extended dechlorination via cis-DCE. These results suggested that the combined application of and the bactrial culture, specially the complete dechlorinating enrichment culture, is practically effective for bioremediation of PCE contaminated soil.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.79-89
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• 2001

Steroid hormones control the expression of many cellular regulators, and a role thor estrogen in mouse oocytes has been well documented. The preovulatory $E_2$increment is generally accepted as the endocrine process regulating induction of in vivo oocyte maturation To address whether the activity of the T-type $Ca^{2+}$ channel is altered by 17 beta-estradiol ( $E_2$), we examined the actions of $E_2$on the calcium channel of mouse oocytes and early embryos. Oocrtes were collected from the oviduct of mice treated with pregnant mare's serum gonadotropin (PMSG) and human choronic gonadotropin (hCG). Whole cell voltage clamp technique and confocal microscopy were used to examine that $E_2$increase intracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{i}$ ) via voltage dependent $Ca^{2+}$ channel (VDC) and estrogen receptor (FSR), and $E_2$concentration by the use of radioimmunoassay (RIA) were examined in mouse. The results obtained were as follows: The peak of $Ca^{2+}$ current induced by $E_2$increased 122% to 1.50$\pm$0.03 nA from 1.23$\pm$0.21 nA (n=15) in the presence of 5 mM extracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{o}$ ). The increased $Ca^{2+}$ current was temporally associated with $Ca^{2+}$ transients. The intracellular $Ca^{2+}$ level increased 207%~30 s following the addition of 1${\mu}{\textrm}{m}$ $E_2$(relative fluorescence intensity: 836.4$\pm$131.2 for control, n=10, 1736.4$\pm$192.0 in the presence of $E_2$, n=10). $E_2$increased amplitude of $Ca^{2+}$ current and [C $a^{2+}$]$_{i}$ . $E_2$-induced $Ca^{2+}$ current and $E_2$concentration in blood were showed difference on the stage of embryo. These results suggest that $E_2$modulate $Ca^{2+}$ channel to increase $Ca^{2+}$ influx.$Ca^{2+}$ influx.

• Journal of Veterinary Clinics
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• v.26 no.6
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• pp.556-562
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• 2009

Skin tumors and mammary gland tumors have been shown to be the most common neoplasia in most of the strains of dogs. The risk for tumor development increases significantly with age and the prevalence and distribution are various according to individual tumors. The aim of this study is to classify histopathologically the skin and mammary gland tumors for recent two years, 2005 and 2006. A total of 128 skin and 240 mammary gland samples of dogs were selected that were submitted to National Veterinary Research and Quarantine Service and Kangwon National University from January 1, 2005 to December 31, 2006. The excised tissue were fixed in 10 percent neutral buffered formalin and processed routinely to paraffin wax. Sections were cut at $3{\mu}m$, stained with haematoxylin and eosin. The slides were examined based on the morphological criteria of M. H. Goldschmidt and W. Misdorp under a light microscope. The age of the dogs ranged from 1 to 19 years with a median of 8.7 years. The mean age of the skin and mammary gland tumors was 7.4 and 9.3 years. 47 (12.8%) were males and 259 (70.4%) were female with a male to female ratio of 0.18. Yorkshire terrier and maltese were more susceptible breeds, accounting for 44.3% of skin and mammary gland tumors. In skin tumors, epithelial, adnexal, and mesenchymal origin tumors were 18 (14.1%), 53 (41.4%), and 57 cases (44.5%), repectively. Among the epithelial, adenexal, and mesenchymal origin tumors, basal cell tumor (8.6%), sebaceous adenoma (15.6%), and histiocytoma (25.0%) were predominant in the incidence rate, respectively. In case of mammary gland tumors, 201 (83.8%) were benign and 39 (16.3%) were malignant with a benign to malignant ratio of 5.15. The most frequent mammary gland tumor was benign mixed tumor (35.0%) followed by mammary adenoma-complex type (31.7%).

• Korean Journal of Veterinary Research
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• v.15 no.2
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• pp.147-152
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• 1975

Although considerable research has been done on the blood picture of the horse, hot-blooded and cold-blooded, little work has been made of the blood picture of the army pack horse, Jeju horse. The object of the present investigation was to make good this deficiency, and to suggest standard for the blood picture of army pack horses kept under the regular military training and the ideal feeding in the heart of a mountain. Blood samples were drawn from the jugular vein through a 15-gauge bleeding needle from 41 males and 28 females, aging 3 to 9 years old. It was taken between seven and nine o'clock in the morning. Animals were handled as quietly as possible to avoid any excitation. No restraint other than a halter was used. Enumeration of erythrocyte, total and differential leukocyte count, determination of hemoglobin in blood, and the value of packed cell volume were male in the usual manner, and erythrocytic constant was calculated by the method of Wintrobe. Erythrocyte count was $7.83{\pm}0.20(4.95{\sim}11.05){\times}10^6/mm^3$(SE). This value was much lower than hot-horses, but slightly higer than the values of cold-horse reported from foreign country. Concentration of hemoglobin in blood was $13.0{\pm}0.33(9.5{\sim}17.8)g/100ml$. This value was much higher than that of cold-horses observed by the other authors, approaching to the values of hot-horses. Packed cell volume was $32.1{\pm}0.92(22{\sim}42)ml/100ml$. This vague was a little higher than that of the other cold-horses. Mean corpuscular volume was $41.5{\pm}1.20(26.6{\sim}59.3){\mu}m^3$. This value matched so well with the other results recorded by various investigators. Mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration were $16.9{\pm}0.43(12.3{\sim}25)$ pg and $41.0{\pm}0.45(29.1{\sim}51.1)g/100ml$, respectively. These values were significantly higher than the values found by the other investigations. Total leukocyte enumeration was $10.5{\pm}0.41(5.6{\sim}17.9){\times}10^3/mm^3$, being considered as normal. And differential leukocyte count of neutrophil was $44.5{\pm}2.23(15{\sim}76)%$, $5,527{\pm}234(2,231{\sim}9,144)/mm^3$, of lymphocyte $50.5{\pm}1.19(19{\sim}77)%$, $4,307{\pm}125(1,456{\sim}11,098)/mm^3$, of monocytel (0~4)%, $105(0{\sim}352)/mm^3$, of eosiophil 3.2(0~14)%, $340(0{\sim}1,232)/mm^3$ and of basophil 0.25(0~3)%, $23(0{\sim}236)/mm^3$. The percentage of the differential count obtained from the present work showed a good agreement with the results of various authors. Of the horses examined monocyte was found from 42 horses, eosinophil from 62 horses and basophil from 10 horses. No significant differences recognized between male and female horses, and the effect of age was not observed between three to nine years old. Judging from the blood picture of the present investigation, it could be stated that the army pack horses on training were kept better than the average farming conditions.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.

• Journal of Life Science
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• v.19 no.11
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• pp.1598-1604
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• 2009

In order to determine chemical components of onion flesh and peel, general nutrients, vitamin C, and total flavonoids were measured. Onion peel showed less moisture (14.3%) and no vitamin C compared to onion flesh. Onion peel contained more amounts of total flavonoids compared to onion flesh. In addition, the inhibitory effects of solvent extracts from onion flesh and peel on $H_2O_$-induced oxidative stress and growth of cancer cell lines (AGS human gastric adenocarcinoma and HT-29 human colon cancer cells) were investigated. Acetone with methylene chloride (A+M) and methanol (MeOH) extracts from onion flesh and peel appeared to significantly reduce the levels of intracellular reactive oxygen species (ROS) (p<0.05) and a greater antioxidant effect was observed in onion peel. Among fractions, 85% aq. methanol showed a higher protective activity against oxidative stress in both flesh and peel and there was no effect in the water and hexane fractions. The growth of cancer cells exposed to medium containing extracts and fractions from onion flesh and peel was inhibited dose-dependently. The growth of AGS was inhibited more in both flesh and peel compared to HT-29, and onion peel was more effective than onion flesh. Among fractions, 85% aq. methanol showed the greatest effect on growth inhibition in both flesh and peel. $IC_{50}$ values of 85% aq. methanol fraction from onion flesh and peel on AGS were 0.04 and 0.03 mg/ml, respectively, while those on HT-29 were 0.23 and 0.04 mg/ml. From our results, 85% aq. methanol fraction had an inhibitory effect against oxidative stress and growth of cancer cells, suggesting that it may contain biological active compounds.

• Journal of Life Science
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• v.19 no.8
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• pp.1073-1080
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• 2009

A number of studies have demonstrated that the regular use of nonsteroidal anti-inflammatory drugs (NSAIDs) can reduce the risks of colorectal, oesophageal and lung cancers. NSAIDs have been shown to exert their anti-cancer effects through inducing apoptosis in cancer cells. The susceptibility of tumor cells to anti-tumor drug-induced apoptosis appears to depend on the balance between pro-apoptotic and anti-apoptotic programs such as nuclear factor kB (NF-kB), phosphatidylinositol 3-kinase (PI3K)-Akt/protein kinase B (PKB) and MEK1/2-ERK1/2 pathways. We examined the effects of pro-survival PI3K and ERK1/2 signal pathways on cell cycle arrest and apoptosis in response to NSAIDs including sulindac sulfide and NS398. We show that simultaneous inhibition of the Akt/PKB and ERK1/2 signal cascades could synergistically enhance the potential pro-apoptotic activities of sulindac sulfide and NS398. Similar enhancement was observed in cells treated with sulindac sulfide or NS398 and 100 ${\mu}$M genistein, an inhibitor of receptor tyrosine kinases (RTKs) that are upstream of PI3K and MEK1/2 signaling. We further demonstrate that NAG-1 is induced and plays a critical role(s) in apoptosis by NSAIDs-based combined treatment. In sum, our results show that combinatorialtreatment of sulindac sulfide or NS398 and genistein results in a highlysynergistic induction of apoptotic cell death to increase the chemopreventive effects of the NSAIDs, sulindac sulfide and NS398.

• Journal of Periodontal and Implant Science
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• v.23 no.3
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• pp.577-594
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• 1993

The ultimate goal of periodontal therapy is the regeneration of the periodontium that have been destroyed as a result of periodontal disease. This study were done in order to determine the healing status of periodontium under Polytetrafluoroethylene and millipore fillter combined with fibrin and the effect of the guided tissue regeneration procedures were performed as follows : 1) flap operation using PTFE membrane(control group) 2) flap operation using PTFE membrane which was fixed with fibrin(experimental group 1) 3) flap operation using millipore filter which was fixed with suture(experimental group II) 4) flap operation using millipore filter which was fixed with fibrin(experimental group II) After 1, 2, 4, 8, 12 weeks, dogs were sacrificed by perfusion technique and tissue block was excised including the tooth and prepared for light microscope with H-E & Masson’s trichrome staining. The result were as follows : In control and experimental group, there is no siginificant difference on epithelial cell down growth within 1st week, but more epithelial cell downgrowth in millipore or millipore combined with fibrin group. In this experiment, there were no significant difference in new cementum and alveolar bone formation whether PTFE membrane was fixed with suture or fibrin. In control and each experimental group, bone maturation appeared in 4 weeks, bone width increased bucco-lingually in control and experimental 1 group especially. Both control group and experimental group showed mild mew cementum formation on root surface and irregular arrangement of collagen fiber at 4 weeks, that showed obvious increased cementum formation at 8 weeks, and that was observed the functional arrangement of collagen fiber between new cementum and new alveolar bone at 12 weeks.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.3
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• pp.267-273
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• 2009

Antioxidative, antimicrobial activities and Raw 264.7 cell viability as cytotoxicity of various solvent extracts from leaf of Eriobotrya japonica Lindl. dried by different methods were investigated for processing as functional ingredient. In DPPH radical scavenging activity, RLE (80% EtOH extract of raw leaf) and FLE (80% EtOH extract of freeze-dried leaf) exhibited strong scavenging effect on $300{\mu}M$ DPPH radical solution (1.71 mg/mL and 2.11 mg/mL for RLE $SC_{50}$ and FLE $SC_{50}$). Also in nitric oxide scavenging activity, RLE and FLE showed strong activities (83.9% and 82.2% in 5 mg/mL sample concentration). Total phenolic compound contents of each extracts were found to be $73.7{\sim}215.4$ mg/g and RLE was showed the highest phenolic compound content. Also, total flavonoid contents were found to be $24.85{\sim}110.3$ mg/g and RLE was showed the highest flavonoid content. In antimicrobial activity, RLE was showed higher growth inhibition effect against all microbial strains. RLE, RLW (hot water extract of raw leaf), and FLW (hot water extract of freeze-dried leaf) exhibited strong antimicrobial activities against MRSA and S. aureus. In measurement of cytotoxicity by MTT assay, Raw 264.7 cell viabilities of 80% EtOH extracts showed better effect than water extracts. Especially viability of RLE was found be over 100% in every tested sample concentration.