• Korean Journal of Microbiology
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• v.22 no.1
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• pp.49-56
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• 1984

Formation and regeneration of protoplasts by Novozym 234 from Kluyveromyces fragilis N100 and Candida pseudotropicalis CBS607 were studied. This enzyme was more effective on cells grown at exponential phase than those at stationary one to convert intact cells into protoplasts. As osmotic stabilizer, ammonium sulphate was suitable for not only protoplast formation but also regeneration in K. fragilis as well as in C. pseudotropicalis. Optimal enzyme concentration was 3mg per ml in K. fragilis and 1~3mg per ml in C. pseudotropicalis, respectively. After the exposure of K. fragilis cells to 3mg per ml of enzyme for 3hr at 30$^{\circ}C$ , approximately 95% of protoplast formation of all observed cells was obgained, while about 100% from C. pseudotropicalis under the same condition was produced. The regeneration frequency of protoplasts by this enzyme was much lower than that by snail enzyme(Glusulase) although Novozym 234 converted cells from above two species into protoplasts free of cell debris effectively, compared with Glusulase. Novozym 234 appears to be suitable for subcellular fractionation to obtain nuclei or other organelles rather than protoplast regeneration.

• The Journal of the Korean Society for Microbiology
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• v.21 no.4
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• pp.407-415
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• 1986

This study investigated whether a correlation exists between environmental physical and biochemical factors and adherence of Candida albicans to human buccal epithelial cells by using normal and UV-irradiated strains. The results were as follows: 1. The percentage of germ tube forming activities of normal Candida albicans was 91.5% and UV-irradiated Candida albicans was 15.0%. The $LD_{50}$ of normal strains in mice were $1.0{\times}10\;cells/ml$, but could not be observed in the UV-irradiated strains even with $1.0{\times}10\;cells/ml$. It demonstrated that the virulence is decreased in the UV-irradiated strain. 2. The adherence of normal Candida albicans to human buccal epithelial cells($166{\pm}29{\sim}207{\pm}17\;cells$/100 epithelial cells) was significantly greater than UV-irradiated Candida albicans($99{\pm}21{\sim}131{\pm}25\;cells$/100 epithelial cells). 3. Candida albicans cultured at $37^{\circ}C$ adhered to buccal epithelial cells($166{\pm}16{\sim}207{\pm}17\;cells$/100 epithelial cells) in greater numbers than cultured at $25^{\circ}C$($80{\pm}15{\sim}143{\pm}22\;cells$/100 epithelial cells). 4. On comparison of the adherence of viable and nonviable(heat-killed) Candida albicans to human buccal epithelial cells, the nonviable Candida albicans demonstrated poorer adherence than viable Candida albicans. 5. Adherence in vitro of Candida albicans to human epithelial cells appeared to be effected by the pH. The adherence ability was maximum increased at pH 7.0($187{\pm}22\;cells$/100 epithelial cells) other than experimental pH. 6. The adherence was proportional to the incubation time and the Candida cell concentration in the suspension. 7. A strong correlation was shown between germ tube forming activity and increased adherence of Candida albicans to human epithelial cells, indicating that germ tube forming activity were responsible for candidal virulence.

• The Korea Journal of Herbology
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• v.29 no.5
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• pp.83-90
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• 2014

Objectives : This research has been done to investigate the anti-inflammatory effect of Coptidis Rhizoma extracts. Method : Coptidis Rhizoma was extracted by $100^{\circ}C$ water. The extract (CC : Extract of Coptis chinensis rhizome) was used to examine its effects on the cell viability of mouse macrophage Raw 264.7 cell line. Also the production of nitric oxide (NO), the c-jun N-terminalkinase (JNK) activation and the production of cytokines such as (IL)-5 were evaluated in lipopolysaccharide (LPS)-stimulated Raw 264.7 cells. After the CC and LPS were applied to Raw 264.7 cells which were cultured for 24 hours, the MTT assay was performed. Result : The CC extracts didn't affect the viability of macrophage cells. However, the extracts inhibited the NO production and the JNK activation significantly in LPS-stimulated macrophage cells treated with 100 and $200{\mu}g/mL$ concentrations. The CC extract, also, impeded the production of inflammation-related factors and cytokines such as KC, VEGF, MCP-1, GM-CSF, IL-$1{\alpha}$, IL-5, IL-6, and IL-12p40 in LPS-stimulated macrophage cells at the concentration higher than $25{\mu}g/mL$. The production of basic-FGF concentration of 50 and $100{\mu}g/mL$, the production of IP-10 at $100{\mu}g/mL$, and the production of IFN-${\gamma}$ at $25{\mu}g/mL$, respectively. Conclusion : The CC prepared using $100^{\circ}C$ water showed the significant anti-inflammatory effect such as the inhibition not only on the production of NO, KC, VEGF, MCP-1, GM-CSF, IL-$1{\alpha}$, IL-5, IL-6, and IL-12p40 in LPS-stimulated macrophage cells at or higher than the concentration of $25{\mu}g/mL$, but also on the JNK activation at 100 and $200{\mu}g/mL$.

• Journal of Nutrition and Health
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• v.36 no.10
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• pp.1022-1029
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• 2003

This study was undertaken to evaluate the antitumor activities of Cordyceps militaris of silkworm pupa (CMP) and silkworm larva (CML), as compared with the effect of cordycepin, an active compound found in Cordyceps militaris. Antiproliferation effect of the test materials were evaluated in the sarcoma-180 cells using the MTT test. For the in vivo study, ICR mice were inoculated i.p. with 1.0 ${\times}$ 10$^{6}$ sarcoma-180 cells/mouse on Day 0, and were again i.p. injected with one of the following substances from Day 1 to Day 10 : saline (control group), 50 mg/kg (CMP50, CML50) ,100 ma/kg (CMP100, CML100), or 200 mg/kg (CMP200, CML200) of Cordyceps militaris water extracts, or 1 mg/kg (C1), 2 mg/kg (C2), or 4 mg/kg (C4) of cordycepin. Pretreatment of the sarcoma-180 cells with 100 mg/ml, 500 mg/ml, and 1000 mg/ml of CML (60.1$\pm$2.5％, 49.8$\pm$3.7％, and 45.4$\pm$0.1％ of the value for untreated control cells, respectively) or CMP (68.3$\pm$2.1％, 55.1$\pm$0.9％, and 51.4$\pm$3.5％ of the value for control cells, respectively) for 48 hrs significantly decreased the survival rate (proliferation) of tumor cells (p<0.05). Body weight of the control mice bearing ascites tumor and injected with saline was 1.4 times of the value for normal animals at day 18. Mice bearing ascites tumor and injected with cordycepin (1, 2, or 4 mg/kg) exhibited a significantly lighter body weight compared with the control mice, while animals injected with CMP or CML (50, 100, or 200 mg/kg) showed a significantly lighter body weight compared with the mice injected with cordycepin. Mice injected with CMP50, CMP100, or CMP200 mg/kg (or CML50, CML100, or CML200 mg/kg) showed a 133％ (or 90％), 80％ (or 62％), and 68％ (or 52％) longer mean survival time, and those treated with C1, C2, or C4 exhibited a 54％, 91％ and 80％ longer survival time compared to the value for control mice injected with saline. These results indicate that the hot-water extracts of Cordyceps militaris of both silkworm pupa and silkworm larva have an anti-proliferation effect of tumor cells as well as the life prolongation effect in mice bearing ascites tumor, which are superior to the activities of cordycepin.

• YAKHAK HOEJI
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• v.34 no.5
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• pp.365-373
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• 1990

The effects of the protein fraction of Panax ginseng on primary cultured chicken embryonic brain cells and DRG cultured with a deficient medium were studied. The protein fraction was further fractionated into four groups according to the molecular weight; larger than 10,000 dalton(fraction A), between 5,000 and 10,000 daltons(fraction B), between 1,000 and 5,000 daltons(fraction C), between 500 and 1,000 daltons(fraction D). All four protein fractions at the concentration of $100\;{\mu}g/ml$ significantly increased the number of the brain cells which promoted the neurite outgrowth. The activity of PDHC in the brain cells was elevated significantly by the protein fraction B at the concentration of $100\;{\mu}g/ml$. It was noted that $100\;{\mu}g/ml$ protein fraction C and D significantly enhanced the synthesis of protein in the brain cells. At the concentration of $100\;{\mu}g/ml$, the protein fraction B enhanced RNA synthesis and the protein fraction A significantly enhanced DNA synthesis in the brain cells. The protein fractions B, C, and D significantly promoted the neurite outgrowth of DRG at the concentration of $100\;{\mu}g/ml$.

• Archives of Pharmacal Research
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• v.18 no.5
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• pp.301-307
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• 1995

In the particulate fraction obtained from PKC-down regulated K562 cells by treatment for 24 h with 200nM TPA, phosphorylation of two proteins with molecular weight, 100 kDa and 23 kDa (designated p100 and p23, respectvely) was depleted and addition of exogenous purified PKC to this fraction failed to testore their phosphorylation. However, in the soluble fraction, all of phosphoproteins abolished by long-term treatment with TPA were restored by exogenously added PKC. Phosphorylation of two proteins was increased by short-term tretment (20 min), and diminished with the persistant exposure to TPA as well as at a concentration as low as 1nM. When K562 cells were treated with 1 nM and 200 nM TPA for 24 h, phosphorylation of p100 was restored with or without exogenous PKC on 2-3day and 6day after removal of treated TPA, respectively. Two-dimensional electrophoresis of phosphoproteins revealed that phosphorylated p100 (pl=5.9) and p66 species were completely absent from the particulate fraction of K562 cells treated with 200nM TPA for 24 h. These results suggest that the interaction of sensitive endogenous substrates, p100 and p23 with the plasma membrane might be regulated by PKC-down regulation without displacement to the cytosol and the interaction of p100 with the membrane might be reveersible.

• Journal of Periodontal and Implant Science
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• v.40 no.3
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• pp.111-118
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• 2010

Purpose: The aim of this study was to investigate whether vitrification in the cryopreservation of periodontal ligament (PDL) cells could be useful for tooth banking. Methods: In step 1, primary cultured human PDL cells were cryopreserved in 100% conventional cryopreservation media and 100% vitrification media (ESF40 media) in different temperatures for 2 weeks. In step 2, a series of modified vitrification formulae named T1 (75% vitrification media + 25% F-media), T2 (50% vitrification media + 50% F-media) and T3 (25% vitrification media + 75% F-media) were used to store PDL cells for 2 weeks and 4 weeks in liquid nitrogen. MTT assay was performed to examine the viability of PDL cells. Results: Maximum cell viability was achieved in cells stored in 100% conventional cryopreservation media at $-196^{\circ}C$ (positive control group) in step 1. Compared to the positive control group, viability of the cells stored in 100% vitrification media was very low as 10% in all test conditions. In step 2, as the percentage of vitrification media decreased, the cell viability increased in cells stored for 2 weeks. In 4-week storage of cells in step 2, higher cell viability was observed in the T2 group than the other vitrification formulae while the positive control group had the highest viability. There was no statistically significant difference in the cell viability of 2-week and 4-week stored cells in the T2 group. Conclusions: These observations indicate 100% vitrification media is not successful in PDL cell cryopreservation. Conventional cryopreservation media is currently the most appropriate media type for this purpose while T2 media would be interesting to test for long-term storage of PDL cells.

• Fisheries and Aquatic Sciences
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• v.8 no.2
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• pp.76-82
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• 2005

We analyzed the paralytic shellfish poisoning (PSP) toxins of the toxic marine dinoflagellate Alexandrium tamarense collected from Dadaepo and Gaduck-do in Busan and from Sujeong-ri in Jinhae Bay, Korea, in April 2003. We also analyzed the PSP toxin of mussels (Mytilus galloprovincialis) and oysters (Crassostrea gigas) collected around Busan and Jinhae Bay. PSP toxin analyses were conducted by high performance liquid chromatography (HPLC). Fifteen cultured A. tamarense isolates contained 2.78 to 57.47 fmol/cell, with nearly identical toxin profiles: major components C2, GTX4; minor components C1, GTX1, NEO; and trace components GTX2, GTX3, STX. PSP toxin contents were 0 to $492\;\mu{g}$ STXeq/100 g in mussels and 0 to $48\;\mu{g}$ STXeq/100 g in oysters. Mussels at Gijang and Sujeong-ri contained the most PSP toxin contents ($492\;\mu{g}\;STXeq/100\;g\;and\;252\;\mu{g}\;STXeq/100\;g,\;respectively$), exceeding the quarantine level ($80\;\mu{g}$ STXeq/100 g). Their dominant toxin components were C2, C1, GTX2, and GTX3; the minor components GTX1, GTX4, GTX5, and NEO were sporadically detected. Phytoplankton contained 0.774 fmol/L seawater and 1.228 fmol/L seawater at Gijang and Sujeong-ri in April. At that time, Alexandrium cells were present in the water column at Gijang at 2,577 cells/mL and at Sujeong-ri at 6,750 cells/mL. Overall, we found the high and similar PSP toxin contents in AZexandrium isolates and mussels, and a correlation between occurrence of toxic Alexandrium cells in the water column and mussel intoxification. High densities of toxic Alexandrium cells in the water column immediately preceded shellfish intoxification at Gijang and Sujeong-ri in April.

• Korean Journal of Veterinary Research
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• v.44 no.3
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• pp.379-387
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• 2004

When an organism is exposed to various toxicants chronically, reactive oxygen species(ROS) are accumulated and eventually result in several biological effects from gene expression to cell death. In the present study we investigated the oxidative damage of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin(TCDD) and/or benzo(a)pyrene (B(a)P) in C100 cells. C100 cells treated with TCDD(30 nM) and B(a)P($3{\mu}M$) underwent diverse oxidative stress as determined through thiobarbituric acid-reactive substances(TBARS) formation, DNA fragmentation, DNA single strand break(SSB) assay, immunohistochemical staining of 8-hydroxy-2'-deoxyguanosine(8-OHdG), and mRNA expressions of antioxidant enzymatic genes such as Cu/Zn-SOD gene, GPx(glutathione peroxidase 5) gene, and catalase gene. Lipid peroxidation in C100 cells was determined through measuing the formation of TBARS. For theat, the cells were pretreated with TCDD(30 nM) and/or B(a)P($3{\mu}M$) for 0.5, 1, 2 and 4 days. TBARS formation was increased in TCDD(30 nM) and B(a)P($3{\mu}M$) and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) and positive control treatment groups comparing to the controls. Mixture treatment induced more DNA fragmentation than the single treatment group at day 6. Also, SSB in all treatment groups was clearly observed when compared with the negative control group. As with the expression of antioxidant enzyme, GPx 5mRNA, B(a)P alone and mixture($30nM\;TCDD+3{\mu}M\;B(a)P$) treatment were higher comparing to those of the negative control and TCDD treatment groups. Our results suggest that exposure of C100 cells to mixture of TCDD and B(a)P leads to significant oxidative damage comparing to the exposures to the individual chemicals. Mechanisms of action are discussed. Additional studies are needed to elucidate the detailed mechanism of mixture-induced toxicity.

• Journal of the Korean Ceramic Society
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• v.40 no.2
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• pp.159-166
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• 2003

The preparation method for composite membranes of high temperature operation above $100^{\circ}C$ for Polymer Electrolyte Membrane Fuel Cells (PEMFCs ) was presented, using perfluorosulfonylfluoride Nafion resin and mordenite, in addition to the physical properties, proton conductivity and single cells performance for it. The composite membranes were fabricated via melting of Nafion resin with various mordenite content. As the increase of mordenite content, at high temperature range, proton conductivity of the composite membrane increased due to the late dehydration rate of existent water in the mordenite. Also, from the result of the current-voltage relationship for single cells under $130^{\circ}C$ operation condition, the composite membrane cell with l0 wt% mordenite content showed better performance than that of the others over the entire current density range. This result indicated that the existent water in the composite membrane with l0 wt% mordenite content was higher than that with the others, thereby maintains its conductivity. Based upon the results of experiments, therefore, a Nafion/mordenite composite membrane prepared by this work is thought to be a satisfactory polymer electrolyte membrane for PEMFC operation above $100^{\circ}C$.