• Journal of Veterinary Clinics
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• v.27 no.3
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• pp.232-239
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• 2010

Two cell lines derived from spontaneous canine mammary gland tumors were established and characterized. Mammary gland tumors from 9 years old pug and 9 years old toy-poodle dogs were collected by aseptic surgical resection and primary culture was performed. The histopathologic examination of tumors revealed adenocarcinoma and complex carcinoma and two dogs died from metastasis of the tumors. The tumor cells were subcultured over 60 times for more than 1 year and morphological consistency maintained. Light microscopic examination, growth curve, doubling time calculation, xenotransplantation to female nude mice, immunohistochemistry for wide spectrum keratin, vimentin, $\alpha$-smooth muscle actin and cytokeratin 8 was performed for characterization. The cell lines exhibited polygonal, elongated cell shape and cytoplasmic bridge and doubling time of 47.1 hrs and 18.6 hrs, respectively. Subcutaneous xenotransplantation to nude mice of the cells produced localized palpable mass within 4 weeks in 4 of 5 and 5 of 5 nude mice, respectively. In immunohistochemical examination one cell line showed strong positive against wide spectrum keratin and cytokeratin 8 and the other cell line showed strong positive against smooth muscle actin and cytokeratin 8. Additional characterization would be possible by investigator's needs and the cell lines may be useful for in vivo and in vitro studies of canine mammary tumor and adjuvant therapies.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2018.04a
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• pp.21-21
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• 2018

본 연구는 북방계 식물에 속하는 두메우드풀[Woodsia ilvensis (L.) R.Br.]의 전엽체 증식 및 포자체 형성을 위한 적정방법을 구명하고자 수행되었다. 식물재료는 경상북도 의성군 일대에서 채취한 후 청주의 일반하우스에 이식하여 재배하였다. 포자 성숙기인 8월에 채종하여 발아시켰으며, 포자로부터 획득한 전엽체를 8주간 계대하면서 실험재료로 사용하였다. 전엽체 300mg을 메스로 다진 후 배지농도 및 종류(Knop, 1/4, 1/2, 1 및 2MS)를 달리하여 8주간 배양하였다. 배양실의 온도는 $25{\pm}1.0^{\circ}C$, 광도는 $30{\pm}1.0{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$(16/8h)로 조절되었다. 연구의 결과, 배지구성물질의 농도가 높았던 2MS배지에서 전엽체의 생체중이 12.4g으로 41배 증가하여 가장 높은 증식율을 나타냈다. 또한 배지구성물질의 농도가 적을수록 생체중의 증가폭도 함께 감소하는 경향을 보였다. 포자체 형성에 적합한 토양조건을 확인하고자, 원예상토, 피트모스, 펄라이트 및 마사토의 비율을 달리한 5종류의 혼합토양을 조성하여 사각분($7.5{\times}7.5{\times}7.5cm$)에 충진하였다. 다음으로 전엽체 1g을 핸드블랜더로 10초간 분쇄 후 충진된 사각분 위에 고르게 분주하여 14주간 재배하였다. 재배환경은 온도 $25{\pm}1.0^{\circ}C$, 광도 $43{\pm}2.0{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$(16/8h) 및 습도 $72{\pm}2.0%$로 조절되었다. 지베렐린 수용액이 포자체 형성에 미치는 영향을 조사하고자, 선행연구에서 선발된 토양조건을 기준으로 침지농도(0, 50, 100, $200mg{\cdot}L^{-1}$)를 조절하여 1시간 침지 후 상기와 동일한 방법으로 재배하였다. 연구의 결과, 원예상토와 펄라이트 또는 마사토를 2:1(v:v)로 혼용한 토양에서 사각분 당 109.5, 128.8개의 포자체가 형성되었다. 한편, 피트모스가 첨가된 처리는 포자체의 형성이 억제되는 경향을 나타냈다. 지베렐린 50과 $100mg{\cdot}L^{-1}$ 수용액에 침지한 처리에서 무처리에 비해 많은 사각분 당 각 177.0, 181.7개의 포자체가 형성되었으며, 생육도 양호하였다.

• Journal of Dental Rehabilitation and Applied Science
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• v.37 no.2
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• pp.88-94
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• 2021

Purpose: The purpose of this study was to evaluate the anti-inflammatory effects of non-thermal atmospheric pressure plasma (NTP) on human gingival fibroblasts (HGFs) for clinical application of periodontal treatment. Materials and Methods: HGFs were treated with Porphyromonas gingivalis (Pg) lipopolysaccharide (LPS). Customized NTP device was developed for periodontal in vitro study. Cell viability was evaluated with cell counting kit-8. The levels of inflammatory cytokines, including interleukin (IL)-8 and 6, were determined by enzyme-linked immunosorbent assay. Results: When NTP was applied, the cell viability did not change significantly, and there was no difference for 6 h and 24h. When Pg LPS was treated to HGFs, the secretion of IL-8 and IL-6 was increased compared to the control group. But when the NTP was applied, the secretion of them was significantly decreased. Conclusion: NTP did not affect cell viability of HGFs. And it inhibited the LPS-induced production of IL-8 and IL-6.

• Journal of Chest Surgery
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• v.39 no.7 s.264
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• pp.511-519
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• 2006

Background: Loss of cardiomyocytes in the myocardial infarction leads to regional contractile dysfunction, and necrotized cardiomyocytes in infracted ventricular tissues are progressively replaced by fibroblasts forming scar tissue. Although cardiomyoplasty, or implantation of ventricular assist device or artificial heart was tried in refractory heart failure, the cardiac transplantation was the only therapeutic modality because these other therapeutic strategies were not permanent. Cell transplantation is tried instead of cardiac transplantation, especially bone marrow is the most popular donated organ. But because bone marrow aspiration procedure is invasive and painful, and it had the fewer amounts of cellular population, the adipose tissue is recommended for harvesting of mesenchymal stem cells. Material and Method: After adipose tissues were extracted from abdominal subcutaneous adipose tissue and intra-abdominal adipose tissue individually, the cellular components were obtained by same method. These cellular components were tried to transformation with the various titers of 5-azacytidine to descript the appropriate concentration of 5-azacytidine and possibility of transformation ability of adipose tissue. Group 1 is abdominal subcutaneous adipose tissue and Group 2 is intra-abdominal adipose tissue-retroperitoneal adipose tissue and omentum. Cellular components were extracted by collagenase and $NH_4Cl$ et al, and these components were cultured by non-induction media - DMEM media containing 10% FBS and inducted by none, $3{\mu}mol/L,\;6{\mu}mol/L,\;and\;9{\mu}mol/L$ 5-azacytidine after the 1st and 2nd subculture. After 4 weeks incubation, tile cell blocks were made, immunostaining was done with the antibodies of CD34, heavy myosin chain, troponin T, and SMA. Result: Immunostaining of the transformed cells for troponin T was positive in the $6{\mu}mol/L\;&\;9{\mu}mol/L$ 5-azacytidine of Group 1 & 2, but CD34 and heavy myosin chain antibodies were negative and SMA antibody was positive in the $3{\mu}mol/L\;&\;6{\mu}mol/L$ 5-azacytidne of Group 2. Conclusion: These observations confirm that adult mesenchymal stem cells isolated from the abdominal subcutaneous adipose tissues and intra-abdominal adipose tissues can be chemically transformed into cardiomyocytes. This can potentially be a source of autologous cells for myocardial repair.

• Tuberculosis and Respiratory Diseases
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• v.66 no.3
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• pp.178-185
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• 2009

Background: Epigallocatechin-3-gallate (EGCG) is the major catechin in green tea, and has shown antiproliferative, antiangiogenic, antimetastatic and cell cycle pertubation activity in various tumor models. Hypoxia can be induced because angiogenesis is insufficient for highly proliferating cancer. Hypoxia-inducible factor-1$\alpha$ (HIF-1$\alpha$) and its downstream target, vascular endothelial growth factor (VEGF), are important for angiogenesis, tumor growth and metastasis. The aim of this study was to determine how hypoxia could cause changes in the cellular phenomena and microenvironment in a non-small cell culture system and to examine the effects of EGCG on a HIF-1$\alpha$ and VEGF in A549 cell line. Methods: A549 cells, a non-small cell lung cancer cell line, were cultured with DMEM and 10% fetal bovine serum. A decrease in oxygen tension was induced using a hypoxia microchamber and a $CO_2-N_2$ gas mixture. Gas analysis and a MTT assay were performed. The A549 cells were treated with EGCG (0, 12.5, 25, 50 ${\mu}mol/L$), and then examined by real-time-PCR analysis of HIF-1$\alpha$, VEGF, and $\beta$-actin mRNA. Results: Hypoxia reduced the proliferation of A549 cells from normoxic conditions. EGCG inhibited HIF-1$\alpha$ transcription in A549 cells in a dose-dependent manner. Compared to HIF-1$\alpha$, VEGF was not inhibited by EGCG. Conclusion: HIF-1$\alpha$ can be inhibited by EGCG. This suggests that targeting HIF-1$\alpha$ with a EGCG treatment may have therapeutic potential in non-small cell lung cancers.

• Restorative Dentistry and Endodontics
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• v.30 no.5
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• pp.409-422
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• 2005

The aim of this study was to identify the bacteria isolated from acute endodontic lesions by cell culture and 16S rDNA sequencing. The necrotic pulpal tissue was collected from 17 infected root canals, which were diagnosed as being either an acute pulpitis or acute periapical abscess. Samples were collected aseptically from the infected pulpal tissue of the infected root canals using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ul of 1 XPBS. The sample solution was briefly mixed and plated onto a BHI-agar plate containing $5\%$ sheep blood. The agar plates were incubated in a $37^{\circ}C$ anaerobic chamber for 7 days. The bacteria growing on the agar plate were identified by 16S rRNA coding gene (rDNA) cloning and sequencing at the species level. Among the 71 colonies grown on the agar plates, 56 strains survived and were identified. In dental caries involving the root canals, Streptococcus spp. were mainly isolated. Actinomyces, Clostridia, Bacteroides and Fusobacteria were isolated in the periapical lesion without dental caries. Interestingly, two new Actinomyces spp. (ChDC B639 and ChDC B631) were isolated in this study. These results showed that there was diversity among the species in endodontic lesions, This suggests that an endodontic infection is a mixed infection with a polymicrobial etiology. These results may offer the bacterial strains for pathogenesis studies related to an endodontic infection.

• Korean Journal of Soil Science and Fertilizer
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• v.17 no.4
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• pp.415-427
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• 1984

This experiment was carried out to investigate the distribution of associative nitrogen fixers, Azospirillum spp. and their nitrogenase activities measured by ARA in the rhizosphers of rice, soybean, and weed grown in the rice paddy field at ear formation stage of rice. Nitrogenase activities produced by Azospirillum spp. enriched from histophere ranged from 16 to 53 n mole/tube/hr.. High nitrogen fixing activities more than 30 n mole/tube/hr. were observed in the histophere of the Echinochloa crus-galli L., Finbristylis miliacea L., and Monochoria vaginalis var.. Nitrogen fixing activities of Azospirillum spp. obtained from single colonies which originated from the rhizoplane of rice (pot-kwang var.), Finbris tylis miliacea L., Monochoria vaginaliz var., Glycine max L. were higher over 100 n mole/tube/hr. than those histophere. Genus of Azopsirillum isoltated from roots of the Graminease (Oriza sativa L., $C_3$-plant, Echinochloa crus=galli L., $C_4$-plant, Cyperus difforuis L.. $C_4$-plant), and Aeschynomene indica L. (Leguminosae, $C_3$-plant) was identified as A. brasilense. However, both strains, A. lipoferum and A. brasilense ($nir^-$ or $nir^+$ strain) were isolated from other plant roots, Both $nir^-$ and $nir^+$ strains of A. brasilense were associated with the same host plant.

• Microbiology and Biotechnology Letters
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• v.8 no.3
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• pp.155-166
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• 1980

Studies have teen undertaken to investigate the degree of microbial contamination in the subsidiary materials which have been known as an important source of microorganisms associated with spoilage of fish sausage and fish paste products. Twenty hinds of food ingredients including starch, spices and condiments, 59 samples in total collected from commercial fish sausage processing plants and supermarket in the period of July to October 1979, were examined for standard plate count, coliform and fecal coliform, mold and yeast, thermoduric microorganisms, aerobic sporeformers (mesophilic and thermophilic), anaerobic sporeformers (mesophilic and thermophilic) and sulfide spoilage anaerobes. The results obtained are summarized as follows. 1. Among the food ingredients examined, corn starch, black pepper, hot pepper, onion, garlic, ginger, beef extract and frank marked high bacterial contamination with general and sporeforming microorganisms. And bacterial content of marked samples were generally higher than that of the samples from plants. 2. The high standard plate count caused by high content of these bacteria like thermoduric, mesophilic or thermophilic sporeforming aerobes. 3. Bacterial content of food ingredients such as black pepper and beef extract being used in plants, and black pepper, hot pepper, onion and garlic from the market were exceeded the bacterial standards being enforced in Japan and U. S. A. 4. Average standard plate count was in the range of 10$^4$to 10$^{5}$ /g for black pepper, wheat flour, onion and garlic collected from plants, and 10$^{5}$ to 10$^{7}$ /g for black pepper, hot pepper, onion and garlic from market. No plate count was observed in pepper essence and coloring material. 5. Coliform organism was detected in starch, black pepper, hot pepper, onion, garlic, ginger and gluten that showed high standard plate but no fecal coliform in the samples except black pepper and hot pepper. 6. Average mold and yeast count was 140 to 460/g for corn starch, wheat flour and black pepper from plants, and 10$^3$/g for black pepper and hot pepper from market. No count was observed in the other ingredients. 7. Sulfide spoilage sporeforming anaerobes boiled for 5 min. at 10$0^{\circ}C$ and incubated at 55$^{\circ}C$ was not detected in all the samples examined.

• The Journal of the Korean Society for Microbiology
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• v.14 no.1
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• pp.27-37
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• 1979

Ninety-five strains of Shigella, 70 of Salmonella paratyphi A, and 230 of Salmonella typhi were tested for their resistance to drugs. Also studied was the inhibition and elimination of drug resistance. All except one strain of Shigella consisted of 79 Sh. flexneri and 16 Sh. sonnei were multiply resistant to chloramphenicol, tetracycline, streptomycin, and splfisomidine. Among them, 70 strains were resistant to ampicillin and carbenicillin, 80 to trimethoprim-sulfamethoxazole, 22 to nalidixic acid, and one to kanamycin, but strain resistant to gentamicin, cephaloridine, and rifampin was not encountered. All strains of S. paratyphi A and S. typhi were susceptible to drugs tested, except sulfisomidine and rifampin, for which all S. paratyphi A were slightly resistant to sulfisomidine and the majority of S. paratyphi A and S. typhi were slightly resistant to rifampin. Approximately 80% of multiply drug-resistant Shigella transferred their resistance to E. coli by conjugation, and the resistance was considered to be mediated by R plasmids. The frequency of transfer of drug resistance varied by donor strains and recipients, but not by selecting drugs. Resistance to nalidixic acid was not transferred by conjugation to the recipients. Drug-resistant Shigella strains successively subcultured in nutrient agar stabs contained clones resistant to drugs and those susceptible to drugs, but the ratio of resistant and susceptible clones varied by strains. The multiply drug-resistant S. typhi and Shigella strains were found to not lose completely their drug resistance by subculture in media. Acriflavine has some effect on the elimination of drug resistance mediated by R plasmids, but the effect varied markedly by strains. Atabrine has no effect among strains tested. The combination of drugs increased the drug actions in majority of cases with synergistic or additive effects.

• KSCE Journal of Civil and Environmental Engineering Research
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• v.29 no.3B
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• pp.303-310
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• 2009

Natural zeolite is widely used as sorbents and bio-media materials because it is cheap as well as it has efficient porous structures and large cation exchange. In this study, the effect of metal cations $(Na^+,\;Ca^{2+},\;Mg^{2+},\;Al^{3+})$ adsorbed to natural zeolite on the microorganism attachment was investigated. Metal-modified zeolites (MMZ) were prepared with 0.01 M, 0.02 M and 0.1 M NaCl, $CaCl_2$, $MgCl_2$ and $AlCl_3$ solutions respectively, which concentrations were equivalent to 10%, 20% and 100% of cation exchange capacity (CEC) of natural zeolite. Pseudomonas putida was used as microorganism which was cultivated in Beef Extract Medium at $26^{\circ}C$. The microorganism attachment to MMZ was increased more than natural zeolite. The amount of bacterial adhesion to MMZ and natural zeolite were $Mg^{2+}>natural>Na^+>Al^{3+}>Ca^{2+}$ under 10% of CEC, $Mg^{2+}>Ca^{2+}>Al^{3+}>natural>Na^+$ under 20% of CEC and $Ca^{2+}>Mg^{2+}>natural>Al^{3+}>Na^+$ under 100% of CEC. Especially, Mg-modified zeolite (Mg-MZ) showed the highest amount of bacterial adhesion, which increased the microorganism attachment 60% higher than natural zeolite under 10% of CEC. However, the amount of bacterial adhesion was decreased as the concentration of metal cations modified to zeolite were increased, showing that the increased amounts were 60% under 10% of CEC, 50% under 20% of CEC and 10% under 100% of CEC in Mg-MZ. Additionally, the effect of $Mg^{2+}$ in solution on the bacterial adhesion was investigated in order to compare it with the effect of $Mg^{2+}$ adsorbed to zeolite. The maximum quantity of bacterial adhesion to Mg-MZ was not different from the amount of microorganism attachment to the natural zeolite when $Mg^{2+}$ solution was added.