• 한국환경성돌연변이발암원학회지
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• 제21권2호
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• pp.89-94
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• 2001

Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

• 한국미생물·생명공학회지
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• 제26권4호
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• pp.316-322
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• 1998

N-$\varepsilon$-aminocaproyl-$\alpha$-D-galactopyranosylamine-sepharose를 담체로 하는 affinity chromatography에 의한 해바라기씨 유래 $\alpha$-galactosidase($\alpha$-D-galactoside galactohydrolase EC 3. 2. 1. 22)의 정제방법과 정제효소에 대한 효소화학적 성질을 규명하였다. N-$\varepsilon$-aminocaproyl-$\alpha$-D-galactopyranosylamine의 흡착제를 합성하여 sepharose에 coupling하였다. 기질 p-nitrophenyl $\alpha$-D-galactopyranoside에 대한 정제효소의 비활성은 291.66 units/mg였고, 조효소와 비교하여 115배의 정제 배율을 나타내었다. 정제효소의 순도는 SDS-polyacryl amide gel전기 영동법 에 의해 단일 band를 나타내었으며, 분자량은 42,000으로 추정되었다. 정제효소의 최적 pH와 온도는 4.5, 55$^{\circ}C$이며, pH 4-5, 30-55$^{\circ}C$의 범위에서 pH와 온도 안정성을 나타내었다. 또한 정제효소는 Ag$^{2+}$, Hg$^{2+}$, CO$^{2+}$의 금속에 의해 70%이상의 저해효과를 나타내었다. 정제효소는 melibiose, raffinose 및 copra galactomannan에 대한 galactose의 유리를 TLC에 의해 확인하였고, 각 기질에 대한 galactose의 가수분해 속도를 HPLC에 의해 비교하였다.

• Asian-Australasian Journal of Animal Sciences
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• 제25권6호
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• pp.895-902
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• 2012

The Korean native chickens (Woorimotdak$^{TM}$, KNC) and commercial broilers (Ross, CB) show obvious differences in meat flavor after cooking. To understand the contribution of protein and peptide for meat flavor, 2-dimensional (2-D) gel electrophoresis and matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry was performed. A total of 16 protein spots were differentially expressed in the breast and thigh meat between the two breeds. A total of seven protein spots were represented by different levels between KNC and CB for breast meat. Among them three protein spots (TU39149, TU40162 and TU39598) showed increases in their expressions in KNC while other four protein spots (BU40125, BU40119, BU40029 and BU39904) showed increases in CB. All nine protein spots that were represented by different levels between KNC and CB for thigh meat showed increases in their expression in KNC. Phosphoglucomutase 1 (PGM 1), myosin heavy chain (MyHC), heat shock protein B1 (HSP27), cytochrome c reductase (Enzyme Q), Glyoxylase 1, DNA methyltransferase 3B (DNA MTase 3) were identified as the main protein spots by MALDI-TOF mass spectrometry. These results can provide valuable basic information for understanding the molecular mechanism responsible for breed specific differences in meat quality, especially the meat flavour.

• Korean Journal of Acupuncture
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• 제37권2호
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• pp.88-96
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• 2020

Objectives : The purpose of this study was to investigate the anti-oxidant and anti-aging effect of the seed of Euphorbia lathyris L. extracted by supercritical CO₂. Methods : Human dermal fibroblast cells dosed with the extract from Euphorbia lathyris L. were harvested and the intracellular proteome was analyzed to examine the expression of proteins related collagen synthesis pathway, metalloproteinases (MMPs), extracellular matrix (ECM)-cell interaction, cytokines, and antioxidant enzymes by 2-dimensional gel electrophoresis. Results : Fatty acid analysis of the extract from Euphorbia lathyris L. showed oleic acid was 84% and linoleic acid was 4.1%. Antioxidative effect was about 53% by beta carotene bleaching assay. In 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis, fifteen protein changes in five mechanisms which were collagen synthesis pathway, MMPs, ECM-cell interaction, cytokines, and antioxidant enzymes were analyzed. Conclusions : This study suggests the supercritical extraction from the seed of Euphorbia lathyris L. could be used as anti-oxidant substances for pharmacopuncture.

• Journal of Microbiology and Biotechnology
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• 제5권5호
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• pp.269-273
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• 1995

A thermotolerable restriction endonuclease. Svil, found in Streptomyces violochromogenes D2-5 was purified. For the purification, streptomycin sulfate and ammonium sulfate precipitation was used. Ph osphocellulose P-ll, DEAE-Cellulose and Sephacryl-S200 HR colum chromatography were also performed. The purified enzyme was found to be homogeneous and the molecular weight of the enzyme estimated by polyacrylamide gel electrophoresis containing 0.1$%$ SDS was about 32, 000 daltons. The recognition sequence and cleavage site of the enzyme were determined to be $5^1$-$TT\downarrow CGAA$-$3^1$ which is the same sequence as that of Asull. Unlike Asull, however, the Svil shows high thermal stability.

• 대한전기학회:학술대회논문집
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• 대한전기학회 2003년도 학술회의 논문집 정보 및 제어부문 B
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• pp.898-902
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• 2003

In this paper we proposes a new technique for identification of breast cancer by classification of proteome pattern generated from 2-D polyacrylamide gel electrophoresis (2-D PAGE) and development of cancer diagnosis system : HABIT. Proteome patterns reflect the underlying pathological state of a human organ and it is believed that the anomalies or diseases of human organs are identified by the analysis or classification of the patterns. Proteome patterns consist of quantitative information of the spots such as their size, position, and density in the proteome image produced from 2-D PAGE, for the Image mining of proteome pattern, SVM(support vector machine) and GA(genetic algorithm) are used to generate a decision model for the identification of breast cancer The decision model was then used to classify an independent set of test proteome patterns into the affecter and unaffecter classes. The proposed technique was tested by actual clinical test samples and showed a good performance of a hit ratio of 90%.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Molecular and Cellular Response to Toxic Substances
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• pp.196-196
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• 2002

p16INK4a tumor suppressor gene transfer in the non-small cell lung cancer cells by transduction of recombinant adenovirus (Ad5CMV-p16) resulted in significant inhibition of cancer cell growth (Anticancer Res., 1998, 18:3257-3261). As a safety concern, we have investigated gene and protein expression after transduction of adenoviral vector (Ad5CMV-p16) in human non-small cell lung cancer (A549) cells by using microarray and 2D gel electrophoresis/ MALDI-TOF.(omitted)

• 한국미생물·생명공학회지
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• 제24권2호
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• pp.197-205
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• 1996

The $\beta$-xylosidase was purified 99- fold from the culture supernatant of Pseudo onas sp. CB-33 by ammonium sulfate precipitation, PEI precipita- tion, DEAE-Sephadex column chromatography, Sephadex G-75 gel filtration chromatography and preparative disc gel electrophoresis. Molecular weight of the enzyme was estimated to be 44,000 by SDS polyacrylamide gel electrophoresis. The enzyme has a pH optimum for activity at 7.0 and is stable over pH 6.5-9.0. The optimal temperature of the enzyme was 45$\circ$C, and its enzymatic activity was completely inactivated at 55$\circ$C for 30 min. Km value of the enzyme for p-nitrophenyl-$\beta$-D-xylopyranoside was calculated to be 4.6 mM. The effect of various reagents on the $\beta$-xylosidase activity was investigated. The enzyme activity was completely inhibited by Hg$^{2+}$, Cu$^{2+}$ and Zn$^{2+}$. The $\beta$-xylosidase was inactivated by tryptophan-specific reagent, N-bromosuccinimide and tyrosine-specific reagent, iodine. The enzyme could degrade xylo-oligosaccharides to xylose and the enzyme was competitively inhibited by xylose. The $\beta$-xylosidase and endoxylanase from Psedomonas sp. CB-33 hydrolized xylan synergically. The purified enzyme also showed $\alpha$-L-arabinofuranosidase activity.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
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• pp.58-58
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• 2006

• 약학회지
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• 제54권1호
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• pp.32-38
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• 2010

The aim of this study was to evaluate the effect of galangin towards hydrogen peroxide-induced DNA damage. The calf thymus DNA and Chinese Hamster Lung (CHL) cells were used to measure 8-hydroxy-2'-deoxyguanosine(8-OH2'dG) as an indicator of DNA oxidative damage using high performance liquid chromatography with electrochemical detection. Hydrogen peroxide in the presence of Fe(II) ion induced the formation of 8-OH2'dG in both calf thymus DNA and CHL cells. The DNA damage effects were enhanced by increasing the concentration of Fe(II) ion and inhibited by galangin. In the single cell gel electrophoresis (Comet assay), galangin and dl-a-tocopherol showed an inhibitory effect in CHL on hydrogen peroxide induced DNA single strand breaks. Galangin showed more potent activity than dl-$\alpha$-tocopherol under our experimental conditions. These results indicate that galangin can modify the action mechanisms of the oxidative DNA damage and may act as chemopreventive agents against oxidative stress.