• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.73-73
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• 2003

본 연구는 돼지 체세포의 clonal cell lines을 효율적으로 확립할 수 있는 배양방법을 제시하기 위하여 실시하였다. 50일령의 돼지태아로부터 섬유아세포를 회수하여 2회 계대배양 후 단일 세포를 96-well plate와 4-well dish에서 배양하여 배양액내의 FBS농도(10, 30, 50%), 첨가제(catalase, $\beta$-mercaptoethanol; BME), 세포의 크기(<16, 16~20, 20<) 및 형태(smooth, rough)에 따른 cell line 확립 효율을 검토하였다. FBS 농도에 따른 clonal line 확립 효율과 PD를 검토한 결과, 효율에 있어서 30% FBS 처리구(5.1%)가 10%(0.3%) 및 50%처리구(2.1%)보다 비교적 높게 나타났으나 유의적 차이는 없었으며, PD의 경우는 10, 30, 50%처리구에서 각각 36.7, 29.4, 26.3 시간으로 FBS 농도가 증가할수록 PD 시간이 유의적으로 짧아졌다(p<0.05). 배양액 첨가제에 있어서 BME 가시 무처리구나 catalase 첨가시보다 유의적으로 높은 효율을 보였으며(11.4%, 3.2%, 3.2%; p<0.05), PD 시간은 짧게 나타났다(23.6, 25.5, 28.1; p<0.05). 그러나 catalase는 cell line 확립 효율에 영향을 미치지 않는 것으로 나타났다. 세포의 크기에 따른 cell line 확립효율은 5.2~8.2%로 크기에 따른 차이는 보이지 않았으며, PD 또한 23.7~27.9 시간으로 세포 크기에 따른 차이는 없었다. 세포의 형태에 따른 세포의 부착율은 smooth(55.8%)구가 rough(73.0%)구보다 유의적으로 낮게 나타났으나(p<0.05), clonal line 확립 효율(7.7%n vs. 6.6%) 및 PD(23.5h vs. 24.0h)에서는 차이가 없었다. 본 연구의 결과, 세포의 형태에 따른 cell line 확립 효율은 큰 차이가 없었으나, 세포 배양액내에 30% FBS와 BME의 첨가로 clonal cell line 확립 효율을 향상시킬 수 있음을 확인할 수 있었다. 또한 cell line 확립 효율이 높아질수록 PD 시간이 짧아지는 경향을 볼 수 있었다.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.181-191
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• 1995

The ability of fibroblasts attach to teeth is of paramount imporance in re-establishing the lost connective tissue attachment after periodontal therapy. Tobacco contains a complex mixture of substances including nicotine. various nitrousamines, trace elements. and a variety of poorly characterized substances. The effects of nicotine on fibroblasts have reported an altered morphology and attachment of fibroblasts to substrates and disturbances in protein synthesis and secretion. This study examined the effect of nicotine, a major component of the particulate phase of tobacco smoke, on human gingival fibroblasts and periodontal ligament cells attachment to tissue culture surfaces and cellular activity of human gingival fibroblasts and periodontal ligament cells. Pooled human gingival fibroblasts made from extraction of 3rd molar were utilized between passage 4 and 5 and plated in 96 well plate at 20,000 cells per well. Cell number were determined using 3-(4,5-dimethylthiazole-2-y)2,5-diphenyltetrazolium bromide(MTI) , which is reflection of mitochondrial dehydrogenase activity. The concentration of nicotine used were 0.025, 0.05, 0.1, 0.2 and $0.4{\mu}M$, the average serum concentration for a smoker being approximately $0.1{\mu}M$. The results were as follows : 1. Attachment effects of nicotine on human gingival fibroblasts and periodontal ligament cells Excepts of $0.4{\mu}M$, the effects on attachment with increasing numbers of cells attaching with increasing nicotine concentrations, compared to control group. But over the 60min, return to control value. 2. The effect of cellular activity on human gingival fibroblasts and periodontal ligament cells. The cellular activity of human gingival fibroblasts and periodontal ligament cells were similar or decrease to control value at 1st incubation day. At 2nd incubation day, 0.05, 0.1, 0.2, $0.4{\mu}M$ concentrations were statistically different from control value on gingival fibroblasts group. But at 3rd incubation day, cellular activities of all experimental group were significantly decrease than control group.

• Korean Journal of Head & Neck Oncology
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• v.17 no.1
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• pp.3-7
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• 2001

Objectives: New photosensitizer 9-hydroxypheophorbide-$\alpha$(9-HpbD-$\alpha$) was derived from chlorophyll in water with peak absorption at 660nm. 9-HpbD-$\alpha$ was tested with 660 nm diode laser for the anticancer effect of photodynamic therapy. Materials and Methods: Human SNU 1041 cells were seeded into 96 well plate at a density of $$ cells/well for 24 hours. Cells were washed with media containing various concentration of 9-HpbD-$\alpha$ ranging from $0{\mu}g/ml\;to\;3.75{\mu}g/ml$. Then, laser treatment was done with 660nm diode laser ($10mW/cm^2$) at various time setting (0, 30, 60, 90, 120 minutes) and with various time interval (0, 1, 4, 6, 18 hours). The treated cells were incubated 48 hours and MTT assay was done to measure the viability of cells. Results: The viability of cells was more than 90% after laser treatment in control group. The viability of cells was decreased with increasing concentration of 9-HpbD-$\alpha$ and laser treatment time in experimental groups. The viability of cells was decreased significantly as the interval time between addition of 9-HpbD-$\alpha$ and laser irradiation was increased. Conclusion: This study shows the anticancer effect of photodynamic therapy using 9-HpbD-$\alpha$ and 660nm Diode laser on carcinoma cell line. 9-HpbD-$\alpha$ is considerd as one of new photo sensitizers in the field of photodynamic therapy.

• Korean Journal of Agricultural Science
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• v.48 no.3
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• pp.557-565
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• 2021

Interest in research on various medicinal plants has increased globally over the last few decades, possibly due to their possible antibacterial and antioxidant activities. The present study was conducted to verify the antioxidant effects, antibacterial activity, and collagen synthesis and cell viability outcomes of adipocytes upon exposure to Abeliophyllum distichum Nakai (AdN). Antibacterial activity was measured through the Disc diffusion method to compare the growth ability of pathogenic microorganisms (E.coli, Salmonella). The absorbance was measured at 560 nm to calculate the active oxygen scavenging ability. Fibroblasts were dispensed in a 96-well plate at a density of 1 × 10⁵ cells·well^-1. The amount of procollagen was measured in each case using a procollagen type 1 C-peptide EIA KIT. The cytotoxicity of the Abeliophyllum distichum Nakai extract against animal adipocytes (Hanwoo backfat cells) was determined using a 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay, a method that measures the conversion of MTS to Formazan by means of mitochondrial dehydrogenases. The concentrations of the samples were made to be 0.0125, 0025, 0.05, 0.1, and 0.5% and all were -completely absorbed into the disc in an incubator at 37℃ for 24 to 36 hours. For the 0.125 mg·disc^-1, effects of Abeliophyllum distichum Nakai on the antioxidant effect, antibacterial activity, and cell viability of adipocytes were found. However, Abeliophyllum distichum Nakai had no effect on collagen synthesis, thus suggesting that AdN extracts may be useful for the prevention and/or treatment of obesity.

• Journal of Life Science
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• v.20 no.10
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• pp.1519-1524
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• 2010

Apigenin (4', 5, 7-trihydroxyflavone), a common dietary flavonoid abundantly present in fruits and vegetables, has shown remarkable anti-proliferative effects against various malignant cell lines. To observe the anti-proliferative effects, oral cavity cancer cell lines, $6{\times}10^3$ cells/well (96 well plate) of KB oral cavity tumor cells were plated and 24 hr later treated with apigenin for one day, after which MTT assay was performed. Apigenin induced cell death in a dose-dependent manner after incubation. Cell viability was significantly decreased in the group treated with 100 ${\mu}M$ apigenin for 24 hr (p<0.05) compared to the control group. To assess apoptosis, the nuclei of KB cells were stained with DAPI. The presence of chromatin condensation in the apigenin treated cells was detected on a fluorescent microscope (${\times}200$). We investigated the in vivo growth inhibitory effects of apigenin on oral cavity cancer KB tumor xenograft subcutaneously implanted in male nude mice. Apigenin was administered to mice by gavage at doses of 25 and 50 mg/kg/day in 0.2ml of PBS. Tumor volume was significantly decreased in 25 and 50 mg/kg apigenin-administration groups compared to the control group. For apoptosis analysis, TUNEL staining was performed. A significant increase in TUNEL positive cells was found in the 25 mg/kg apigenin administration group compared to the non- apigenin administration group. Histopathological changes were not observed. These results indicate that apigenin inhibits oral cavity cancer cell growth through the induction of apoptosis.

• Journal of Life Science
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• v.15 no.5 s.72
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• pp.715-725
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• 2005

Oral streptococci are major constituents of dental plaques, and their prevalence is closely linked with various pathologic symptoms, such as dental caries. To develop natural anticaries agent, we prepared 309 kinds of plant extracts from 215 species of edible or medical plants, and antibacterial activity of the extracts against Streptococcus mutans JC-2 were evaluated based on 96 well microtiter plate assay and disk paper method, subsequently. Among the tested plant extracts, Ailanthus altissima, Paeonia lactiflora, Rubus phoenicolasius, Aralia continentalis, Quercus acutissima, Persicaria hydropiper and Agrimonia pilosa extracts showed strong antimicrobial activity. Determination of minimal inhibitory concentration (MIC) of the selected seven plant extracts showed that Ailanthus altissima, Persicaria hydropiper and Quercus acutissima extracts ($MIC=25\∼30[\mu}g/ml$) has potential as a source of natural anticaries agents.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.4
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• pp.273-278
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• 2015

The purpose of this study was to evaluate the in-vitro efficacy of PDT using red light emitting diode (LED) with Radachlorin for biofilm inhibition of clinical Candida albicans isolates. The suspensions containing C. albicans at $9{\times}10^8CFU/mL$ were prepared on yeast nitrogen base containing 5% glucose. The biofilm formation was grown for 3 h after seeding suspensions each 100 ul on a 96-well plate and then supernatant was discarded. Each well was treated with $0.39{\mu}g/mL$ from $50{\mu}g/mL$ concentrations of Radachlorin on adherent biofilm. After a 30-minute incubation, light was irradiated for 30, 60, or 90 minutes using the following light source of wavelength 630 nm LED, at energy densities of 14, 29, and $43J/cm^2$. Afterwards, all supernatant was removed and dried. Adherent cells were stained with safranin O and dried. The cell viability was measured using a microplate reader at 490 nm. Also, a fluorescent signal on C. albicans was observed by saturation of a photosensitizer. In conclusion, a significant inhibition of 72.5% was observed to C. albicans on biofilm at the Radachlorin dose of $50{\mu}g/mL$ with 630 nm LED. The Photosensitizer (Radachlorin) was adequate at 30 minuttes for C. albicans. Overall, the results showed that inhibition of biofilm formation was Radachlorine dose-dependent. The results suggest that PDT, using Radachlorin with 630 nm LED, is able to decrease biofilm formation of C. albicans.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.39-42
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• 2012

A new method for the high throughput screening of antagonistic substances against rice pathogens using rice leaf explants was developed. This method can be used to confirm the activities of any compound or mixture suppressing rice bacterial blight (BB) before field tests. Xanthomonas oryzae pv. oryzae (Xoo) culture medium was distributed in 96 well plates with equally sized explants and the active compounds were added to the wells. The strength suppressing BB was converted into an area percent of the lesion on the rice explants. The explants under BB suppressing activity remained uninfected maintaining their actual green color, while infected explants exhibited pale yellow-colored lesions. Based on the results, this method seems to be faster and easier, dose-dependent, and can be performed all-at-once with a small amount of unspecified compounds. This method also has the potential to be applied to inspection activities for the suppression of other waterborne crop diseases.

• Restorative Dentistry and Endodontics
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• v.27 no.6
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• pp.600-611
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• 2002

본 연구의 목적은 클로로필린이 치주인대 세포 생활력에 미치는 영향을 평가하는데 있다. 치주인대 세포는 건강한 사람에서 발치한 소구치의 치주인대 조직으로부터 채취하여 배양하였다. 비교 기준을 설정하기 위하여, HBSS 내에서 $37^{\ circ}C$로 보관한 치주인대 세포수의 50%가 생존하는데 소요되는 시간을 MTT 분석법에 의해 측정하였다. 그 결과는 6시간이었다. HBSS에 클로로필린 10, 100, 500 nM이 각각 첨가된 3군의 실험 보존액과 F-medium, ViaSpan, Likorol 등 모두 6군의 실험 보존액을 96-well plate에 접종한 후, 치주인대 세포를 동일한 수로 분주하였다. 이를 6시간동안 보관한 후, 각 실험군 보존액에서의 세포 생활력을 MTT 분석법으로 측정하였다 또한, HBSS와 F-medium 및 클로로필린 500 nM이 첨가된 HBSS군의 세포들에 대해 유식세포 계측을 시행하여 각각의 세포주기를 분석하였다. 실험 결과, 클로로필린 500 nM이 첨가된 HBSS에서 보관한 세포들이 가장 높은 생활력을 나타내었으며, 클로로필린이 첨가되지 않은 HBSS에 비해서 유의하게 양호한 세포 생활력 유지 능력을 보였다. 클로로필린으로 처치한 세포들은 클로로필린의 농도가 커짐에 따라 세포 생활력이 증가하는 양상을 나타내었다. 유식세포 계측 결과, HBSS와 F-medium 및 클로로필린 500 nM이 첨가된 HBSS에서 보관한 세포의 77~80%가 G0-G1 단계의 세포 주기로 측정되어, 대부분의 세포들이 안정된 세포 대사 상태를 보이는 것으로 나타났다. 결론적으로, 클로로필린 처치는 치주인대 세포의 생활력 유지에 도움이 되는 것으로 사료된다.

• Korean journal of food and cookery science
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• v.25 no.2
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• pp.189-198
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• 2009

This study was conducted to analyse middle school students' perceptions of seafood as well as their eating experiences and favorite cooking methods for seafood. It also examined the frequency and use of seafood on school lunch menus. Questionnaires were distributed to 739 students in 5 middle schools in the Busan area. On a scale of 5.00, the average scores for perceptions toward seafood were as follows: 3.67 for health & nutrition, 3.53 for taste, 2.53 for smell, 2.77 for convenience. The male students had significantly_(p<0.05) higher perception scores for health & nutrition, taste, and smell than the female students. Among 38 types of seafoods, the highest percentages for eating experiences included seaweed_(99.2%), anchovies_(99.1%), fried fish paste_(98.6%), shrimp_(98.4%), crab_(98.2%), squid_(97.9%), sea mustard_(97.8%), mackerel_(97.7%), dried squid_(97.3%), and hair tail_(96.9%). However, the majority of students had eaten a limited amount of seafood. The degree of seafood intake at home was significantly_(p<0.01) higher than at school. The students' favorite cooking methods for seafood were in the order of grilling, stir-frying, steaming, deep-fat frying, and soup. However, the middle school lunch programs primarily served as soup, seasoning, stir-fried, braised, deep-fat fried, and grilled, respectively. The main reasons for leaving seafood on a plate were inconvenience due to the fishbones_(62.7%), tasteless_(45.4%), and disliking the smell_(35.7%). Improvements that students wanted for seafood on school lunch menus included removal of fishbone for convenience_(67.0%), taste_(52.0), and increased variety of seafood types_(38.2%).