• Korean Journal of Clinical Laboratory Science
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• v.49 no.4
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• pp.390-394
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• 2017

Unexpected antibody screening and identification tests are highly important in the prevention of hemolytic transfusion reactions. Therefore, it is highly recommended to perform unexpected antibody screening test in all transfusion candidates. Here, the frequency and distribution of unexpected antibodies identified in Jeju for the past 3 years were evaluated. Between Jan 2014 and Dec 2016, unexpected antibody screening test was performed for 10,360 sera of transfusion candidates in Jeju general hospital using a column agglutination method with the Ortho BioVue system (Ortho-clinical Diagnostics, Raritan, NJ, USA). Eighty-seven (0.84%) of 10,360 cases that underwent unexpected antibiotics screening showed positive results. Among them, unexpected antibodies were identified in 41 cases (0.40%). Unidentified antibodies were detected in 8 cases (19.51%) and autoantibodies were detected in 3 cases (7.32%). The anti-E antibody included in warm antibodies were detected most frequently in 8 cases (19.51%); 6 cases (14.63%) of anti-E + anti-c antibody and 3 cases (7.32%) of $anti-Le^a+anti-Le^b$. $Anti-Le^a$ and $anti-Le^b$ antibodies were detected in 2 cases (4.88%), respectively. The anti-D, $anti-Di^a$, $anti-Fy^b$, $anti-Jk^a$, $anti-Jk^b$, anti-M and anti-P1 were detected in 1 case (2.44%). Complex antibodies were detected in 1 case (2.44%) in anti-C+anti-D and anti-E+anti-c+$anti-Jk^b$, respectively. In this study, we analyzed the frequency and distribution of unexpected antibodies in one general hospital for the past 3 years. However, there has been a general increase in multicultural families and foreign workers in Jeju, and it would be a meaningful study to compare the frequency and distribution of unexpected antibodies.

• Journal of Life Science
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• v.27 no.2
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• pp.202-210
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• 2017

As an effort to find a potential biopreservative, we isolated bacterial strains producing bacteriocin from fermented foods. A strain was finally selected and characteristics of the bacteriocin were investigated. The selected strain was identified as Bacillus subtilis E9-1 based on the 16S rRNA gene analysis. The culture supernatant of B. subtilis E9-1 showed antimicrobial activity against Gram-positive bacteria. Subtilisin A, ${\alpha}$-chymotrypsin, trypsin and proteinase K inactivated the antimicrobial activity, which means its proteinaceous nature, a bacteriocin. The bacteriocin activity was fully retained at the pH range from 2.0 to 8.0 and stable at up to $100^{\circ}C$ for 60 min. Solvents such as ethanol, isopropanol and methanol had no effect on the antimicrobial activity at the concentration of 100% but acetone and acetonitrile reduced the activity at up to 100% concentration. Cell growth of four indicator strains was dramatically decreased in dose-dependent manner. Listeria monocytogenes was the most sensitive, but Enterococcus faecium was the most resistant. Bacillus cereus and Staphylococcus aureus showed the medium sensitivity. The bacteriocin showed its antimicrobial activity against B. cereus and L. monocytogenes via bactericidal action. The number of viable cells of L. monocytogenes started to reduce after addition of bacteriocin to the minced beef. The bacteriocin was purified through acetone concentration, gel filtration chromatography and RP-HPLC. The whole purification step led to a 6.82 fold increase in the specific activity and 6% yield of bacteriocin activity. The molecular weight of the purified bacteriocin was determined to be 3.3 kDa by MALDI-TOF/TOF mass spectrometry.

• Journal of Life Science
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• v.27 no.11
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• pp.1276-1289
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• 2017

This study isolated and purified the antimicrobial peptide McSSP-31 from an acidified hemocyte extract of a Mytilus coruscus. The antimicrobial peptide was purified by using a $C_{18}$ reversed-phase high-performance liquid chromatography (HPLC). The peptide was determined to be 3330.549 Da by matrix assisted-laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS). The N-terminus of a 14 amino-acid sequence was identified as P-S-P-T-R-R-S-T-S-R-S-K-S-R by Edman degradation method. The acquired sequence showed a 93% similarity with the sperm-specific protein Phi-1, which is from M. californianus. The identified open-reading frame (ORF) of peptide was 306 bp encoding 101 amino acids, which was analyzed by rapid amplification of cDNA ends (RACE), cloning and sequencing analysis. We compared the full sequence with other known proteins that reveal the sperm-specific protein Phi-1 (93.5%) of M. californianus. Synthesized antimicrobial peptide (McSSP-31) showed antibacterial activity against gram-positive bacteria including B. subtilis, S. mutans, S. aureus and gram-negative bacteria including E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa and fungi, C. albicans. Also, synthesized peptide showed strong antibacterial activity against antibiotic-resistant strains, including S. aureus. The cytotoxicity of the peptide was determined by using the HUVEC human cell line. The peptide did not exhibit any significant cytotoxic effects on the normal human cell line, and it had very low hemolytic activity with flounder hemoglobin. The results demonstrated that peptide purified from the hemocyte of a M. coruscus exhibits antibacterial activity against various bacteria and has the potential to be an alternative antibiotic agent.

• Journal of Life Science
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• v.22 no.11
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• pp.1456-1462
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• 2012

Sorafenib is a multikinase inhibitor and an oral anticancer drug approved for the treatment of patients with advanced renal cell carcinoma and those with unresectable hepatocellular carcinoma. The purpose of this study was to develop an efficient method of the determination of sorafenib in human plasma using tandem mass spectrometry coupled with liquid chromatography (LC/MS/MS) and validate the method by the guidelines of the Korean Food and Drug Administration (KFDA). Plasma samples ($100{\mu}l$) were added with chlorantraniliprole as an internal standard and then mixed with the 0.1% formic acid-containing extraction solution composed of isopropyl alcohol and ethyl acetate (1:4, v/v). After centrifugation, the supernatant was concentrated at $45^{\circ}C$ under negative pressure and centrifugal force. The residue was reconstituted with a mobile phase and injected into the HPLC instrument using a reverse phase Waters XTerra$^{TM}$ C18 column (particle size $3.5{\mu}m$). Liquid chromatography was carried out within the run time of 5 min using a mobile phase composed of buffer (0.1% formic acid and 10 mM ammonium formate), methanol, and acetonitrile (1:6:3, v/v/v). The analytes were monitored by tandem mass spectrometry in the multiple reaction monitoring method programmed to detect sorafenib at 'm/z 465.2 ${\rightarrow}$ 252.5' and chlorantraniliprole at 'm/z 484.4 ${\rightarrow}$ 286.2' with positive electrospray ionization mode ($ES^+$). The result showed the proper linearity ($r^2$ > 0.99) over the range of 2,000-5,000 ng/ml with good accuracy (90.7-103.9%) and precision (less than 10%). The newly developed method using LC/MS/MS was validated by the guideline of KFDA and identified as more sensitive compared to the previous methods.

• Journal of the Korean Vacuum Society
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• v.17 no.4
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• pp.322-330
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• 2008

The properties of discharge, luminance, and spectroscopy are investigated in a longitudinal and transverse discharge fluorescent lamps with tube of outer diameter 4 mm. The sample lamps are prepared to be three kinds of gas composition such as mercury lamps of Ne(95%)+Ar(5%)+Hg(2 mg), the mercury-free lamps of Xe 100% and Ne+Xe(4%). The gas pressure is in the range of $5{\sim}300\;Torr$. In the mercury lamps, the longitudinal discharge having a positive column is high in luminance and efficiency, while the transverse discharge is no luminance at all. In the Xe-lamps, the transverse discharge shows relatively good in efficiency as compared with the longitudinal discharge which has a high discharge voltage and a low luminance and efficiency. In the transverse discharge of relatively high efficiency, a pure Xe(100%) gas discharge has a higher efficiency than the mixture gas of Ne+Xe(4%). Through these experiments, the properties of mercury and xenon lamps are verified. In the mercury lamps, the longitudinal discharge of tubular fluorescent lamps is high in luminance and efficiency, while the transverse discharge of flat panel fluorescent lamps are low in luminance efficiency. In the mercury-free lamps, the flat fluorescent lamps of transverse discharge having a high pressure ${\sim}100\;Torr$ with the pure Xe-gas are verified to be suggestable.

• Food Science and Preservation
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• v.24 no.1
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• pp.93-99
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• 2017

Collagen synthesis is decreased and matrix metalloproteinase-1 (MMP-1) levels are increased in naturally aged human skin, and these alterations cause changes such as skin wrinkling and decreased elasticity. As a part of our ongoing search for bioactive ingredients, MMP-1 inhibitory and type-1 procollagen synthesis inducing activities of aqueous methanolic extract of manufactured gambir product from Uncaria gambir were investigated in in vitro bioassay systems. In addition, total phenolic contents were quantified using a spectrophotometric method. Among tested samples, 40% MeOH eluate from 80% methanolic extract of manufactured U. gambir using open column chromatography packed with Diaion HP-20 resin showed significant MMP-1 inhibitory activities with an $IC_{50}$ value of $15.6{\pm}1.3{\mu}g/mL$. Furthermore, type-1 procollagen synthesis promoting property of 40% MeOH eluate ($IC_{50}$ value; $6.9{\pm}0.7{\mu}g/mL$) from 80% methanolic extract of manufactured gambir was higher than other eluates. Additionally, the present investigation revealed that 40% MeOH eluate of manufactured gambir product contained a high level of total phenolic compounds. The result suggests a distinct relationship between anti-wrinkle activity and total phenolic contents, and manufactured gambir product could be considered a new effective source of natural bioactive ingredients. Systematic investigation of manufactured gambir product will be performed for further development of its biological properties.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.4
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• pp.502-508
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• 2009

Antimicrobial activity of Sargassum thunbergii was determined by paper disc assay and minimum concentration inhibitor (MIC) test. A water extract of S. thunbergii did not show the antimicrobial activity, but an ethanol extract of S. thunbergii (SHE) inhibited Serratia liquefaciens, Salmonella Typhimurium, Pseudomonas aerogenosa and all of the tested gram-positive bacteria at 4 mg/mL. Especially, Bacillus subtilis, Clostridium perfringens and Listeria monocytogenes were susceptible to SHE. As the results of MIC test, SHE inhibited the growth of B. subtilis, Staphylococcus aureus and Listeria monocytogenes at concentration of $0.1{\sim}0.3%$, and inhibited C. perfringens at 0.01%. In the thermal and pH stability test for SHE, antibacterial activities of SHE were maintained when the SHE was treated at $121^{\circ}C$ for 15 minutes or under pH $2{\sim}8$. SHE was partitioned in the order of n-hexane, chloroform, ethyl acetate and butanol. As the results of the MIC test for each obtained fraction, no fraction exhibited higher antibacterial activity than that of the crude SHE. However, a mixture of chloroform, ethylacetate and ethanol fractions showed higher antibacterial activity than SHE.

• Journal of Food Hygiene and Safety
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• v.29 no.2
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• pp.131-140
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• 2014

The objective of this study was to develop a simultaneous method of 8 penicillin antibiotics including amoxicillin, ampicillin, cloxacillin, dicloxacillin, nafcillin, oxacillin, penicillin G and penicillin V in meat using LC-MS/MS. The procedure involves solid phase extraction with HLB cartridge and subsequent analysis by LC-MS/MS. To optimize MS analytical condition of 8 compounds, each parameter was established by multiple reaction monitoring in positive ion mode. The chromatographic separation was achieved on a $C_{18}$ column with a mobile phase of 0.05% formic acid and 0.05% formic acid in acetonitrile at a flow rate of 0.2 mL/min for 20 min with a gradient elution. The developed method was validated for specificity, linearity, accuracy and precision in beef, pork and chicken. The recoveries were 71.0~106%, and relative standard deviations (RSD) were 4.0~11.2%. The limit of detection (LOD) and the limit of quantification (LOQ) were 0.003~0.008 mg/kg and 0.01~0.03 mg/kg, respectively, that are below maximum residue limit (MRL) of the penicillins. This study also performed survey of residual penicillin antibiotics for 193 samples of beef, pork and chicken collected from 9 cities in Korea. Penicillins were not found in all the samples except a sample of pork which contained cloxacillin (concentration of 0.08 mg/kg) below the MRL (0.3 mg/kg).

• Journal of Life Science
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• v.28 no.11
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• pp.1301-1315
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• 2018

We purified an antimicrobial peptide from the acidified hemocyte extract of Mytilus coruscus by $C_{18}$ reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was 4041.866 Da based on matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS) and the 25 amino acids of the N-terminus sequence were identified. Comparison of this sequence of the purified peptide with the N-terminus sequences of other antimicrobial peptides revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We also identified a 312 bp open-reading frame (ORF) encoding 103 amino acids based on the obtained amino acid residues. The nucleotide sequence of this ORF and the amino acid sequence also revealed 100% identity with the mytilin B precursor of Mytilus coruscus. We synthesized two antimicrobial peptides with an alanine residue in the C-terminus, and designated them mytilin B1 and B2. These two antimicrobial peptides showed antimicrobial activity against gram-positive bacteria, including Bacillus cereus and Streptococcus parauberis (minimal effective concentration, MECs $41.6-89.7{\mu}g/ml$), gram-negative bacteria, including Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, and Vibrio ichthyoenteri (MECs $7.4-39.5{\mu}g/ml$), and the fungus Candida albicans (MECs $26.0-31.8{\mu}g/ml$). This antimicrobial activity was stable under heat and salt conditions. Furthermore, the peptides did not exhibit significant hemolytic activity or cytotoxic effects. These results suggest that mytilin B could be applied as alternative antibiotic agent, and they add to the understanding of the innate immunity of hard-shelled mussels.

• Herbal Formula Science
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• v.31 no.1
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• pp.1-10
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• 2023

Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.