• Korean Journal of Food Science and Technology
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• v.38 no.2
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• pp.176-182
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• 2006

Sensitive and specific monoclonal antibody (MAb) was produced from hybridoma (1H11-5) obtained by fusion of myeloma cell (V653) and spleen cell isolated from mouse immunized sulfamthazine (SMZ)-HG-KLH. Direct competitive ELISA was developed for rapid detection of SMZ in milk samples using MAb against SMZ with optimized conditions between MAb and SMZ-HG-HRP conjugate, and applicable conditions for analysis of milk samples were established. Detection range of immunoassay was 0.1 to 100 ppb. Recoveries from spiked raw milk and processed milk samples averaged 82.1-120.7 and 82.1-97.1%, respectively.

• Korean Journal of Food Science and Technology
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• v.35 no.4
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• pp.556-560
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• 2003

It was determined whether the sandwich ELISA using specific anti-CP4 EPSPS polyclonal and monoclonal antibodies, developed in the previous study, could be applied to detect GM soybean or not. The soybeans (47 imported and 20 domestic soybeans) were analyzed by a sandwich ELISA. The results of imported soybeans were divided into two groups which were high contents $(39.1{\pm}13.5\;{\mu}g/g,\;n=33)$ and low contents of CP4 EPSPS $(2.6{\pm}1.2\;{\mu}g/g,\;n=14)$. The ratio of GM in imported soybeans was about 70.2%. One the other hand, the contents of CP4 EPSPS in domestic soybeans was very low $(0.9{\pm}0.5\;{\mu}g/g,\;n=20)$ which determined to be non-GM soybeans. In case of soybean sprouts, the contents of CP4 EPSPS in soybean sprouts were different between GM and non-GM soybean sprout. The CP4 EPSPS in cotyledon of GM soybeans sprout was higher than that in root hair. The contents of CP4 EPSPS in soybeans sprout of domestic soybeans were very low. Thus, it was possible to determine that the soybeans sprout was made of GM or non-GM soybeans. Also, PCR experiment showed that the sandwich ELISA was accurate to distinguish the soybeans to be GM or non-GM. These results showed the sandwich ELISA could determine the soybeans were GM or non-GM, rapidly and simply.

• Korean Journal of Food Science and Technology
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• v.44 no.4
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• pp.488-492
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• 2012

Two strains, traditionally referred to as rock flower (Bawhi-kot) and buckwheat flower (Memil-kot or Chile-Kot), were isolated from stored traditional soy sauce and were identified by using the 18S ITS1/4 region sequences. The rock flower strain showed 99% of similarity with Cladosporium sp. and buckwheat flower strain was 99% identical with yeast Sterigmatomyces halophilus. Both strains were tentatively named Cladosporium sp. NK1 and Sterigmatomyces halophilus NK2, respectively. The optimal growth pHs and temperatures of both strains in a YPD broth medium were in the range of pH 5.0 to 7.0 and 22 to $27^{\circ}C$. Both strains were able to grow in more than 20% of NaCl. In the enzyme activity assay, high protease activity of Cladosporium sp. NK1 and S. halophilus NK2 were obtained in YPD containing 10% of NaCl. High amylase activities of both stains were in 15% and 5% of NaCl, respectively. Lipase activity was, however, not detected in both strains.

• Journal of Life Science
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• v.24 no.9
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• pp.995-1000
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• 2014

The studies of marine viruses in terms of viral isolation and detection have been limited due to the high mutation rate and genetic diversity of marine viruses. Of the modern methods currently used to detect marine viruses, serological methods based on enzyme-linked immunosorbent assay (ELISA) are the most common. They depend largely on the quality of the antibodies and on highly purified suitable antigens. Recently, a new experimental system for using viral capsid protein as an antigen has been developed using the yeast surface display (YSD) technique. In the present study, the capsid protein gene of the red-spotted grouper nervous necrosis virus (RGNNV) was expressed and purified via YSD and HA-tagging systems, respectively. Two regions of the RGNNV capsid protein gene, RGNNV1 and RGNNV2, were individually synthesized and subcloned into a yeast expression vector, pCTCON. The expressions of each RGNNV capsid protein in the Saccharomyces cerevisiae strain EBY100 were indirectly detected by flow cytometry with fluorescently labeled antibodies, while recognizing the C-terminal c-myc tags encoded by the display vector. The expressed RGNNV capsid proteins were isolated from the yeast surface through the cleavage of the disulfide bond between the Aga1 and Aga2 proteins after ${\beta}$-mercaptoethanol treatment, and they were directly detected by Western blot using anti-HA antibody. These results indicated that YSD and HA-tagging systems could be applicable to the expressions and purification of recombinant RGNNV capsid proteins.

• Molecules and Cells
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• v.40 no.3
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• pp.222-229
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• 2017

Adipose-derived stem cells (ADSCs) were previously considered to have an anti-inflammatory effect, and Interleukin-$1{\beta}$ ($IL-1{\beta}$) was found to be a pro-inflammatory factor in chondrocytes, but the mechanism underlying ADSCs and $IL-1{\beta}$ is unclear. In this study, we investigate whether P2X7 receptor (P2X7R) signalling, regulated by microRNA 373 (miR-373), was involved in the ADSCs and $IL-1{\beta}$ mediated inflammation in osteoarthritis (OA). Chondrocytes were collected from 20 OA patients and 20 control participants, and ADSCs were collected from patients who had undergone abdominal surgery. The typical surface molecules of ASDCs were detected by flow cytometry. The level of nitric oxide (NO) was determined by Griess reagent. Concentrations of prostaglandin E2 (PGE2), interleukin 6 (IL-6), matrix metallopeptidase 3 (MMP-3) were detected by enzyme-linked immunosorbent assay (ELISA). The expressions of IL-6, MMP-3, miR-373 and P2X7R were determined by real-time polymerase chain reaction (PCR), and Western blot was used to detect the protein expression of P2X7R. The typical potential characters of ADSCs were verified. In chondrocytes or OA tissues, the miR-373 expression level was decreased, but the P2X7R expression was increased. $IL-1{\beta}$ stimulation increased the level of inflammatory factors in OA chondrocytes, and ADSCs co-cultured with $IL-1{\beta}$-stimulated chondrocytes decreased the inflammation. OA chondrocytes transfected with the miR-373 inhibitor increased the inflammation level. The miR-373 mimic suppressed the inflammation by targeting P2X7R and regulated its expression, while its effect was reversed by overexpression of P2X7R. $IL-1{\beta}$ induced inflammation in OA chondrocytes, while ADSCs seemed to inhibit the expression of P2X7R that was regulated by miR-373 and involved in the anti-inflammatory process in OA.

• Journal of Preventive Medicine and Public Health
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• v.40 no.4
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• pp.285-290
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• 2007

Objectives : Cases of human brucellosis in Korea have recently increased due to the increasing incidence of bovine brucellosis. The authors conducted this study to elucidate the status of brucellosis through seroepidemiologic study. Methods : We selected our study population from a high risk group. We conducted a questionnaire survey and obtained blood samples to determine the seroprevalence of brucellosis antibodies for 10 days in February, 2005. The titers of brucellosis were measured by the combination of standard tube agglutination test (STA) and enzyme-linked immunosorbent assay (ELISA) test. Results : Our study subjects comprised 1,075 cases: 971 livestock workers, 51 veterinarians, and 53 artificial inseminators. In the STA test, 27 cases (2.5%) had titers of greater than or equal to 1:20. Of 1,068 cases (7 cases were excluded due to previous brucellosis), 7 cases of brucellosis were diagnosed with titers of 1:160, giving a seroprevalence of brucellosis of 0.66%. The seroprevalence in the male group was 0.95%, and that of livestock workers, veterinarians, and artificial inseminators was 0.52%, 4.17%, and 0.00%, respectively. The Spearman's correlation coefficient between the positive rate of bovine brucellosis per capita and household and human brucellosis was 0.806 and 0.744, respectively. The concordance rate between the Korea National Institute of Health and the Gyeongsangbuk-do Institute of Health and Environment by the STA and ELISA tests was 94.7% and 100.0%, respectively. Conclusions : The study results indicated in higher seroprevalence rate among veterinarians than among livestock workers and artificial inseminators. Because veterinarians may be exposed to this high risk, effective working guidelines for veterinarians to guard against brucellosis must be developed. Moreover, more extensive epidemiologic research for laboratory workers and meat handlers is needed.

• The Korean Journal of Mycology
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• v.26 no.1 s.84
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• pp.8-15
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• 1998

The present research was undertaken to investigate the activity of ligninolytic enzymes and the decolorization capability of some dyes with Irpex zonatus BN2, isolated from nature and identified. For the assay of enzyme activities, the isolate did not produce lignin peroxidase (LiP) and veratryl alcohol oxidase (VAO), but laccase and manganese dependent peroxidase (MnP). While the activity for MnP was low $(61.6\;nmol/mg{\cdot}protein)$, its laccase activity was very high $(1185.9\;nmol/mg{\cdot}protein)$. Moreover, laccase had appeared earlier than MnP. When the isolate was incubated with each dye for 10 days, the decolorization rates of azo dyes, such as orange II, orange G, tropaeolin O and congo red were 98.0%, 97.4%, 99.0% and 95.3%, respectively. In case of heterocyclic dyes, eosin Y, toludine blue, methyl blue and azur B were 97.4 %, 98.7%, 99.9% and 94.0% respectively. Finally the results of triphenylmethane dye such as basic fuchsin, malachite green and crystal violet were 98.5%, 95.7% and 99.4%, respectively. The results suggest that laccase of Irpex zonatus BN2 should be played an important role in the decolorization of the dyes.

• Tuberculosis and Respiratory Diseases
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• v.75 no.5
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• pp.205-209
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• 2013

Background: We investigated whether prunetin significantly affects tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced MUC5AC mucin gene expression, production, inhibitory kappa B ($I{\kappa}B$) degradation and nuclear factor kappa B (NF-kB) p65 translocation in human airway epithelial cells. Methods: Confluent NCI-H292 cells were pretreated with prunetin for 30 minutes and then stimulated with TNF-${\alpha}$ for 24 hours or the indicated periods. MUC5AC mucin gene expression and mucin protein production were measured by reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The effect of prunetin on TNF-${\alpha}$-induced degradation of $I{\kappa}B$ and translocation of NF-${\kappa}B$ p65 was investigated by western blot analysis. Results: We found that incubation of NCI-H292 cells with prunetin significantly inhibited mucin production and down-regulated the MUC5AC gene expression induced by TNF-${\alpha}$. Prunetin inhibited TNF-${\alpha}$-induced degradation of $I{\kappa}B$ and translocation of NF-${\kappa}B$ p65. Conclusion: This result suggests that prunetin inhibits the NF-${\kappa}B$ signaling pathway, which may explain its role in the inhibition of MUC5AC mucin gene expression and production regulated by the NF-${\kappa}B$ signaling pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.39 no.10
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• pp.1418-1424
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• 2010

This study was performed to assess the antioxidant activities and whitening effects of Sorbus commixta HEDL cortex on melanin synthesis. The whitening effects of Sorbus commixta HEDL cortex water extracts were examined by in vitro mushroom tyrosinase assay and B16BL6 melanoma cells. We assessed inhibitory effects of Sorbus commixta HEDL cortex water extracts on expression of melanogenic enzyme proteins including tyrosinase, tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2 (TRP-2) in B16BL6 cells. Inhibitory effects of Sorbus commixta HEDL cortex onto free radical generation were determined by measuring DPPH and hydroxyl radical scavenging activities. Our results indicated that Sorbus commixta HEDL cortex water extracts effectively inhibited free radical generation. In DPPH radical scavenging activity, Sorbus commixta HEDL cortex water extracts had a potent anti-oxidant activity in a dose-dependent manner. They significantly inhibited tyrosinase activity in vitro and in B16BL6 melanoma cells. Also, Sorbus commixta HEDL cortex suppressed the expression of tyrosinase, TRP-1 and TRP-2 in B16BL6 melanoma cells. These results show that Sorbus commixta HEDL cortex inhibited melanin production on the melanogenesis. The underlying mechanism of Sorbus commixta HEDL cortex on whitening activity may be due to the inhibition of tyrosinase activity and tyrosinase, TRP-1, TRP-2 expression. We suggest that Sorbus commixta HEDL cortex may be contain new natural active ingredients for antioxidant and whitening cosmetics.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1308-1316
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• 2008

Propionibacterium acnes is known to playa pivotal role in the pathogenesis of acne vulgaris. CBT-SL5 is one of the antimicrobial peptides from Enterococcus faecalis SL5, and it has shown antimicrobial activity against P. acnes. The aim of this study was to investigate the anti-inflammatory effect of CBT-SL5 on the inflammation induced by P. acnes in cultured human keratinocyes. Cultured human keratinocytes derived from neonatal foreskin were treated with heat-killed P. acnes to induce inflammation, and then various concentrations of CBT-SL5 were added to the P. acnes-treated keratinocytes. The mRNA expression and protein secretion of interleukin (IL)-8, an inflammation marker, was analyzed by real-time reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. We also analyzed the nuclear factor-kappa B (NF-$\kappaB$) p65 translocation by performing immunofluorescent staining. P. acnes treatment up regulated the IL-8 mRNA expression in the keratinocytes, and this was brought about through both toll-like receptor (TLR)2 and TLR4. At the concentrations of 10, 50, and 100 ng/ml, CBT-SL5 significantly down regulated the P. acnes-induced IL-8 mRNA expression and protein production (p<0.05). At 6 hand 12 h of the treatment, CBT-SL5 significantly suppressed the P. acnes-induced IL-8 mRNA expression. Secretion of IL-8 protein was significantly reduced at 24 h. The functional inhibitory activity of CBT-SL5 was shown by CBT-SL5 suppressing the P. acnes-induced NF-$\kappaB$ translocation from the cytoplasm to the nucleus. These results demonstrated that CBT-SL5 suppressed the P. acnes-induced IL-8 expression in keratinocytes. Therefore, CBT-SL5 may be a novel anti-inflammatory treatment for acne.