• Biomolecules & Therapeutics
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• v.26 no.5
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• pp.474-480
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• 2018

Most angiogenesis assays are performed using endothelial cells. However, blood vessels are composed of two cell types: endothelial cells and pericytes. Thus, co-culture of two vascular cells should be employed to evaluate angiogenic properties. Here, we developed an in vitro 3-dimensional angiogenesis assay system using spheroids formed by two human vascular precursors: endothelial colony forming cells (ECFCs) and mesenchymal stem cells (MSCs). ECFCs, MSCs, or ECFCs+MSCs were cultured to form spheroids. Sprout formation from each spheroid was observed for 24 h by real-time cell recorder. Sprout number and length were higher in ECFC+MSC spheroids than ECFC-only spheroids. No sprouts were observed in MSC-only spheroids. Sprout formation by ECFC spheroids was increased by treatment with vascular endothelial growth factor (VEGF) or combination of VEGF and fibroblast growth factor-2 (FGF-2). Interestingly, there was no further increase in sprout formation by ECFC+MSC spheroids in response to VEGF or VEGF+FGF-2, suggesting that MSCs stimulate sprout formation by ECFCs. Immuno-fluorescent labeling technique revealed that MSCs surrounded ECFC-mediated sprout structures. We tested vatalanib, VEGF inhibitor, using ECFC and ECFC+MSC spheroids. Vatalanib significantly inhibited sprout formation in both spheroids. Of note, the $IC_{50}$ of vatalanib in ECFC+MSC spheroids at 24 h was $4.0{\pm}0.40{\mu}M$, which are more correlated with the data of previous animal studies when compared with ECFC spheroids ($0.2{\pm}0.03{\mu}M$). These results suggest that ECFC+MSC spheroids generate physiologically relevant sprout structures composed of two types of vascular cells, and will be an effective pre-clinical in vitro assay model to evaluate pro- or anti-angiogenic property.

• Journal of the Korean Society of Food Science and Nutrition
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• v.46 no.7
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• pp.896-902
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• 2017

In this study, we investigated citrus Kombucha (CK) produced by three different bacteria strains (Gluconacetobacter xylinus, Gluconacetobacter medellinensis, and Gluconobacter oxydans; named as CK-MOX) identified from traditional Kombucha. During fermentation, the pH level of CK-MOX was gradually reduced, and total acidity slightly increased. Antioxidant activity, measured by DPPH, ABTS, and oxygen radical absorbance capacity assays, markedly increased after fermentation. Moreover, fermented CK-MOX (Day15) exhibited anti-proliferative and anti-migratory activities against EJ human bladder carcinoma cells. Western immunoblot assays showed that treatment with CK-MOX significantly up-regulated phospho-extracellular signaling kinase (ERK) levels. To distinguish whether or not up-regulation of phospho-ERK is the cause or effect, we investigated the viability of EJ cells in the presence of U0126, a mitogen activated protein kinase/ERK kinase 1/2 inhibitor. Pre-treatment with U0126 rescued cells from CK-MOX-induced cell death, which indicates phospho-ERK may be a key regulator in the mechanism of CK-MOX-induced apoptosis of EJ bladder cancer cells. In conclusion, CK-MOX, fermented by a defined composition of bacterial starters, shows antioxidant capacity and anti-cancer activity against EJ bladder cancer cells.

• Korean Journal of Microbiology
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• v.39 no.3
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• pp.135-140
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• 2003

Bacillus sp. secretes high levels of extracellular enzymes into the culture medium. The signal recognition particle (SRP) and the SRP receptor play a central role in targeting pre secretory proteins to the translocase. By the analysis of the DNA microarray of B. lentimorbus WJ5, it was detected that WJ5m12, antifungal activity deficient mutant induced by gamma radiation, had a down-regulated expression of the SRP receptor gene (B. subtitis srb homologue, srbL). To determine the relationship of SRP receptor to antifungal activity, srbL of B. lentimorbus WJ5 was amplified by PCR and ligated into pQE30 vector, and then transferred into WJ5m12. The transformant, WJ5m12::srbL, recovered the antifungal activity. From the 2-DE analysis, the several presecretory proteins accumulated in the mutant cell and decreased to a level of the wild type in WJ5m12::srbL. It seems that the srbL could play an important role in the secretion of the antifungal activity related proteins of B. lentimorbus WJ5.

• Korean Journal of Veterinary Service
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• v.37 no.4
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• pp.253-261
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• 2014

Porcine circovirus disease (PCVD) is a major problem of swine industry worldwide, and diagnosis of PCV2, causal agent of PCVD, has been doing in clinical laboratories of pig disease by polymerase chain reaction (PCR) methods. But the PCR analyses have a serious problem of misdiagnosis by contamination of DNA, in particular, from carryover contamination with previously amplified DNA or extracted DNA from field samples. In this study, an uracil DNA glycosylase (UNG)-based direct PCR (udPCR) without DNA extraction process and DNA carryover contamination was developed and evaluated on PCV2 culture and field pig samples. The sensitivity of the udPCR combined with dPCR and uPCR was same or better than that of the commercial PCR (cPCR) kit (Median diagnostics, Korea) on PCV2-positive serum, lymph node and lung samples of the pigs. In addition, the udPCR method confirmed to have a preventing ability of mis-amplification by contamination of pre-amplified PCV2 DNA from previous udPCR. In clinical application, 170 pig samples (86 tissues and 84 serum) were analysed by cPCR kit and resulted in 37% (63/170) of positive reaction, while the udPCR was able to detect the PCV2 DNA in 45.3% (77/170) with higher sensitivity than cPCR. In conclusion, the udPCR developed in the study is a time, labor and cost saving method for the detection of PCV2 and providing a preventing effect for DNA carryover contamination that can occurred in PCR process. Therefore, the udPCR assay could be an useful alternative method for the diagnosis of PCV2 in the swine disease diagnostic laboratories.

• The korean journal of orthodontics
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• v.39 no.3
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• pp.177-184
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• 2009

Objective: The aim of this study was to investigate the occurrence rate of bacteremia following toothbrushing with toothpastes composed of several antibacterial agents and compare the results with the conventional oral hygiene maintaining methods in orthodontic patients. Methods: This clinical study included 100 adult orthodontic patients who were divided into 4 groups. Each group comprised of 25 patients, wearing fixed orthodontic appliances. In the first group, bacteremia was assessed after toothbrushing without using any toothpaste. In the second group, a 0.2% chlorhexidine gluconate mouthrinse was used before brushing with no toothpaste. In the third group, subjects brushed with a commonly used toothpaste which did not include an additional antimicrobial agent. The fourth experimental group used toothpaste which included tea tree oil, clove oil, peppermint oil and bisabolol as antimicrobial elements. Pre- and post-brushing blood samples were obtained using a strict aseptic technique. All samples were microbiologically evaluated using blood culture bottles. Results: Toothbrushing in orthodontic patients yielded to an increase in the occurrence rate of bacteremia when using normal toothpaste or no toothpaste at all. Conclusions: The use of chlorhexidine mouthwash before toothbrushing, and brushing with antimicrobial toothpaste did not show a statistically significant difference in preventing the occurrence of bacteremia (p > 0.05).

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.8
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• pp.1289-1295
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• 2014

The aim of this study was to select the most suitable starter cultures for production of fermented sausages. A total of 27 strains isolated from Korean fermented foods and natural substances were characterized with respect to their physicochemical properties in a fluid (submerged) model system modified according to the special conditions of fermented sausages. Three of these strains were pre-selected for testing as potential cultures based on their ability to grow fast and initiate rapid acidification. The selected strains were identified by API and partial sequence analysis of 16S rRNA. The results exhibited sequence similarity to known sequences of Staphylococcus warneri, Staphylococcus epidermidis and Lactobacillus plantarum. Among them, relatively good growth properties and nitrite reduction activities were detected for S. epidermidis and L. plantarum and low pH values and high total acidities were observed in the model system fermented with these isolates compared with reference strains.

• Journal of Family Resource Management and Policy Review
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• v.20 no.3
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• pp.95-113
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• 2016

This study investigates the potential for multi-cultural youths, given their innate bilingual and dual culture, to be trained as the competent ODA professionals. For the purpose of collecting information, we organized two different kinds of FGI in 2015. The first FGI was conducted with a group of 10 expert participants from August 17 to September 25. The second FGI was conducted four times, from October 5 to October 23, with 13 adolescents from multi-cultural families from Russia, Mongolia, Vietnam, India, Indonesia, Japan, and China. The semi-structured interview items were passed through a formal review process of the Institutional Ethics Committee. Furthermore, with the participants' pre-consent to recording, all recorded data were categorized through the transcription process. The results are as follows. While the potential for nourishing bilingual and dual cultural capabilities are high for the adolescent children of multi-cultural families, the expert group also emphasized the negative aspects of these capabilities being suppressed due to the relatively oppressed family environment. The expert group therefore suggests the following: building parent awareness of ODA human resources, developing an educational career transition roadmap, and providing institutional incentives for ODA. In addition, to some extent, the researchers were able to detect some positive self-esteem among the adolescent children of multi-cultural families as a result of the bilingual and dual cultural competencies required to be ODA personnel. In sum, the research provides insight into the need for national social support for the adolescent children of multi-cultural families, so that their strengths are not extinguished, in order that they might grow up as competent ODA personnel.

• Korea Journal of Hospital Management
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• v.22 no.3
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• pp.144-156
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• 2017

Purpose: Crisis is inevitable to every organization and therefore, successful crisis management is critical to the organizations' survival and prosperity. With the understanding, this study aims to draw propositions for successful crisis management of hospitals when facing infectious disease outbreak. For the purpose, a case of a small and medium sized hospital's experience of crisis management during 2015 Middle East Respiratory Syndrome outbreak was analyzed. Methodology/Approach: The detailed internal circumstances and experiences of the hospital during the MERS outbreak were identified by in-depth interview as well as the extensive material review, and analyzed under the view of the theories of accident, error, and crisis in relation of organization management Findings: Overall, nine propositions are drawn by the phase of crisis. In pre-crisis phase, for example, 'the hospital preparedness has positive influence on the effective responding to the crisis'. In detection phase, 'the mindfulness of the hospital organizations' as well as the individuals' has positive influence on detecting the crisis signals'. In crisis phase, for example, 'improvising naturally occurs in crisis by the unknown disease, therefore, a component site supervisor coordinating such improvision is important'. Lastly, in post-crisis phase, 'successful crisis responding experience facilitates the positive hospital culture'. Practical implication: From the experience of a small and medium size hospital, it is suggested that proactive system approach oriented by safety is beneficial for effective crisis management.

• YAKHAK HOEJI
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• v.51 no.6
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• pp.415-423
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• 2007

This study was to evaluate the genotoxic effects of airborne particulate matters using single cell gell elec trophoresis (comet assay) in A549 human lung carcinoma cells. The total suspended particulate (TSP) was collected on back-up filter in Chuncheon, Kangwon Do, South Korea from April, 2003 to February, 2005. The concentrations of TSP, B(a)p and most of heavy metals seemed to be higher in spring and winter, and lower in summer. And they showed higher concentration in the commercial areas and the residential area having more traffics than in the rural area. It was found that A549 cells interacting with the organic extract of TSP showed more DNA single-strand breaks compare to untreated cells. The genotoxicity of the organic extract of TSP was increased with the pre-treatment of S-9 mixture during the culture or with the treatment of endonuclease after cell lysis. The DNA damage by the organic extract of TSP was higher in winter and the commercial area than in summer and the rural area. This study suggests that TSP, heavy metals and B(a)P analyzed showed significant variation depend on the seasons and the areas which are correlated with the DNA damage evaluated by Comet assay, indicating that genotoxic biomarker is useful for toxicological evaluation of air quality.

• The Korea Journal of Herbology
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• v.23 no.3
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• pp.53-60
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• 2008

Objectives : Oxidative stress has a critical role in neurodegenerative diseases. In this study, we investigated the antioxidant and neuroprotective effects of the ethanolic extract of Banryong-hwan (BRHE) in SH-SY5Y cells and primary rat mesencephalic dopaminergic neurons. Methods : To assess the antioxidant effects, we carried out 1,1-diphenyl-2-picrylhydrazyl(DPPH) free radical scavenging assay, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)(ABTS) radical cation decolorization assay, and determination of total polyphenolic content. We evaluated the effect of BRHE treatment on neuroprotection against 6-hydroxydopamine(6-OHDA) toxicity using thiazolyl blue tetrazolium bromide(MTT) assay, nitric oxide(NO) assay, reactive oxygen species(ROS) assay in SH-SY5Y cells and tyrosine hydroxylase(TH) immunocytochemistry in primary rat mesencephalic dopaminergic neurons. Results : BRHE showed IC50 values of 328.10 ${\mu}g/mL$ and 43.12 ${\mu}g/mL$ in DPPH assay and in ABTS assay, respectively. Total polyphenolic content was 180.76 ${\mu}g/mL$. In SH-SY5Y cells, BRHE significantly attenuated the toxicity induced by 6-OHDA at the concentrations of 25-100 ${\mu}g/mL$ pre- and post- treatment in MTT assay. While 6-OHDA increased the NO and ROS contents, BRHE decreased them in a dose dependent manner. Moreover, in primary dopaminergic neuron culture, BRHE significantly protect-ed the dopaminergic cell loss against 6-OHDA toxicity up to 136% at the concentration of 75 ${\mu}g/mL$. Conclusions : These results demonstrate that BRHE has neuroprotective effect against 6-OHDA induced neurotoxicity through decreasing NO and ROS generation.