• Title/Summary/Keyword: Normal phase HPLC

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The Antimicrobial Characteristics of McSSP-31 Purified from the Hemocyte of the Hard-shelled Mussel, Mytilus coruscus (참담치(Mytilus coruscus) 혈구(hemocyte)에서 분리한 McSSP-31의 항균 특성 분석)

  • Oh, Ryunkyoung;Lee, Min Jeong;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Joo-Won;Park, Jung-Youn;Seo, Jung-Kil;Kim, Dong-Gyun
    • 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.

A study of relationship between stomach cancer and selenoproteins in Korean human blood serum (한국인 혈청에서의 셀레노 단백질과 위암과의 상관관계 연구)

  • Park, Myungsun;Pak, Yong-Nam
    • Analytical Science and Technology
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    • v.28 no.6
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    • pp.417-424
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    • 2015
  • In this study, the relationship between selenoprotein concentrations in blood and stomach cancer have been searched for Korean. The concentration of each selenoprotein in blood serum was analyzed and the correlation between the concentration and stomach cancer was studied to find a potential for using Selenium as a biomarker. In concentration determination, a simple calibration curve method was used with the monitoring of m/z 78 without the use of solid phase extraction. This is a lot more simple than the method using SPE with post column isotope dilution. The result obtained from the analysis of CRM BCR-637, 72.20±3.35 ng·g−1, showed similar value of reference value (81±7 ng·g−1). The total concentration of Se for the controlled group, cardiovascular patients group, was 105.70±21.20 ng·g−1. This value was the same as normal healthy person reported earlier. Each selenoprotein concentration of GPx, SelP and SeAlb was 26.12±7.84, 65.15±14.50, 14.43±6.99 ng·g−1, respectively. The distribution of each selenoprotein was 24.7%, 61.6%, and 13.7%, which was similar to the normal person. The result of stomach cancer patients, the total concentration of Se was 76.11±28.12 ng·g−1 and each concentration of GPx, SelP and SeAlb was 15.41±9.01, 50.83±17.91, and 9.87±5.21 ng·g−1, respectively. The total and each selenoprotein concentration level showed significant decrease for the stomach cancer patients. The level of decrease was 41.0% for GPx, 22.0% for SelP, and 31.6% for SeAlb. However, the distribution of each selenoprotein was not much different. Either total Selenium or each selenoprotein could be used as a possible index for the diagnosis of cancer. However, in age group study, it is shown that young age group (30's-40's) did not show much difference.

Influence of UV Irradiation Procedures on the Concentration of Vitamin $D_{3}$ and 25-Hydroxyvitamin $D_{3}$ in the Liver and Skeleton of Broiler Chicks (자외선 조사방법이 육계 병아리의 간장과 골격중 Vitamin $D_{3}$ 및 25-Hydroxyvitamin $D_{3}$ 농도에 미치는 영향)

  • ;;M.F.Holick
    • Korean Journal of Poultry Science
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    • v.21 no.3
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    • pp.157-168
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    • 1994
  • This study was carried out to elucidate the time course variation of vitamin $D_{3}$ (V$D_{3}$) and 25-hydroxyvitamin $D_{3}$ [25(OH)$D_{3}$] contents in the liver and skeleton of 3-wk old broiler chicks when treated with different UV irradiation procedure. Day-old Hubbard chicks were fed vitamin D deficient diet for 3 wk and exposed to medium wave ultraviolet(UVB) light with different irradiation procedures. Procedure I was 30 min continuous irradiation(O.204 mJ /$cm^{2}$) and Procedure II was three seperate 10 min irradiation with 12 h intervals, and Procedure III was three seperate 10 min irradiation with 24 h intervals. The liver and skeleton samples were collected at 10 different times between 0000~2400 h after the last irradiation. The V$D_{3}$ and 25(OH)$D_{3}$ fractions wereseparated by Sep-Pak silica cartridge and the concentrations were determined by normal phase HPLC. The mean content of V$D_{3}$ in the liver of the birds treated by Procedure II was 6.68 ng /g, which was higher than 5.60 and 5.30 ng /g from Procedure I and Ill, respectively(P<.O5). With regard to the effect of elapsed time after UVB irradiation on the V$D_{3}$ concentration of the liver, 96 h treatment showed the highest value(13.08 ng/g)(P<.05). There was a significant(P$D_{3}$ were significantly(P$D_{3}$ in the skeleton of tibia and femur, there were no significant differences among Procedure I, II and III, but significant differences were found among those from various elapsed time after irradiation, The highest value was shown at 96 h(O.99 ng /g) treatment, and interaction between irradiation procedure and elapsed time was not significant. With regard to the mean content of 25(OH)$D_{3}$ in bone, the Procedure II(18.79 ng /g) and III(17.73 ng /g) showed higher values than Procedure I did (P<.05), and the 12 h elapsed time showed the highest value(31.17 ng /g) among 10 treatments (P<.05), however, there was no significant interaction between exposing procedure and elapsed time. In conclusion, the Procedure II would he more desirable than Procedure I or III to produce more V$D_{3}$ and 25(OH)$D_{3}$ in the liver and skeleton of birds by exposing to the UVB light. Also, it was verified that 25(OH)$D_{3}$ increases more quickly than V$D_{3}$ in both tissues tested and is utilized more quickly to recover from the rickets of chicks.

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Antioxidant Compounds and Activities of Methanolic Extracts from Oat Cultivars (품종별 귀리 메탄올 추출물의 항산화 성분 및 항산화 활성)

  • Ham, Hyeonmi;Woo, Koan Sik;Lee, Byongwon;Park, Ji-Young;Sim, Eun-Yeong;Kim, Byung Joo;Lee, Choonwoo;Kim, Si-Ju;Kim, Wook Han;Lee, Junsoo;Lee, Yu-Young
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1660-1665
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    • 2015
  • Oats (Avena sativa L.) were evaluated in recent years as a promising crop for increasing the nutritional quality of foods, due to their abundance in many bioactive compounds. The objectives of this study were to determine antioxidant compounds and antioxidant activities of nine oat cultivars and investigate the relationships between antioxidant compounds and antioxidant activities in oat cultivars. The contents of total polyphenolics and vitamin E in oat cultivars were analyzed by spectrophotometric methods and normal phase HPLC, respectively. 2,2'-Azino-bis-(3-ethylbenzothiazoline- 6-sulphonic acid) (ABTS) radical and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities, reducing power, and chelating effects were used to compare the relative antioxidant activities of oat cultivars. In this study, total polyphenolic and vitamin E contents of oat cultivars ranged from 83.41 to 130.59 mg gallic acid equivalents/100 g and 0.54 to 1.90 mg/100 g, respectively. High levels of ABTS radical (116.14 mg Trolox equivalent antioxidant capacity (TEAC)/100 g) and DPPH radical (38.58 mg TEAC/100 g) scavenging activities and reducing power ($A_{700}=0.922$) were found in Daeyang. Significant correlation was evaluated between total polyphenolic contents and their antioxidant activities, namely ABTS and DPPH radical scavenging activities and reducing power. A low correlation was observed between vitamin E contents and their antioxidant activities. The results of this study could have an impact on oat consumption by enhancing consumer awareness of the health benefits of oats.