• Title/Summary/Keyword: rapid screening

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The Rapid Detection of Antioxidants from Safflower Seeds (Carthamus tinctorius L.) Using Hyphenated-HPLC Techniques (Hyphenated-HPLC 기술을 활용한 홍화씨의 항산화 성분 분석)

  • Kim, Su-Jin;Kim, Sang-Min;Kang, Suk-Woo;Um, Byung-Hun
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.414-419
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    • 2010
  • Hyphenated-HPLC techniques combine the separation power of HPLC with the structural and bioactivity information provided by NMR, ESI/MS, and an on-line antioxidant screening system. The major advantages over the traditional off-line techniques are rapidity and efficiency. In this study, we used hyphenated HPLC techniques including online HPLC-ABTS, LC-NMR, and LC-MS todirectly identify the major antioxidants of safflower (Carthamus tinctorius L.) seeds. The results demonstrated that the major antioxidant compounds from on-line HPLC-ABTS analysis were identified as 8'-hydroxyarcgenin-4'-O-$\beta$-D-glucoside, N-(p-coumaroyl) serotonin, and N-feruloylserotonin. Among them, N-feruloylserotonin accounted for almost 50% of the ABTS radical scavenging activity of the total extract. The results demonstrate that HPLC hyphenated techniques can be used to rapidly screen and structurally identify antioxidants from crude plant extracts.

Establishment of the Physicochemical and Radiological Database of Raw Materials and By-Products in Domestic Distribution (국내 유통중인 원료물질 및 공정부산물의 물리화학적 및 방사선적 특성 데이터베이스 구축)

  • Lim, Chung-Sup;Lim, Jong-Myoung;Park, Ji-Young;Chung, Kun Ho;Kim, Chang-Jong;Chang, Byung-Uck;Ji, Young-Yong
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.14 no.4
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    • pp.331-341
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    • 2016
  • To evaluate the physicochemical and radiological properties of raw materials and by-products in domestic distribution, about 220 samples with 16 species were prepared. We measured the energy spectrum and the chemical content, such as U, Th, and K, using a $LaBr_3$ scintillation detector and ED-XRF. In addition, HPGe detector was used to analyze the radioac-tivity of $^{234}Th$, $^{234}mPa$, and $^{214}Bi$ in uranium decay series and $^{228}Ac$, $^{212}Pb$, and $^{208}Tl$ in thorium decay series, and $^{40}K$. The correlation between characteristic variables, such as the count rate in several ROIs, chemical content, and radioactivity, was assessed to infer the radioactivity of natural radionuclides through a rapid screening method. Based on the results, a characteristic database for raw material and by-product in domestic distribution was established and it will provide useful information in the analysis procedure and improve the accuracy and reproducibility in the analysis of natural radionuclides.

Distinguishing the Korean Silage Corn Varieties through Development of PCR-Based SNP Marker (SNP마커 개발을 통한 사료용 옥수수 품종판별)

  • Kim, Sang Gon;Lee, Jin-Seok;Bae, Hwan Hee;Kim, Jung-Tae;Son, Beom-Young;Baek, Seong-Bum
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.37 no.2
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    • pp.168-175
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    • 2017
  • Single nucleotide polymorphisms (SNP) markers allow rapid screening of crop varieties in early growth stages. We developed a modified SNP PCR procedure for assaying SNPs in maize. For SNP marker development, we chosen 200 SNP sites from MaizeGDB database, and designed two base pair mismatch primers based on putative SNP site of B73 genome sequence. PCR products size was from 200 to 500 bp or was not shown in the case of SNP site existing in Korean silage corns. Using previously discovered 16 primer sets, we investigated distinctness of 50 silage F1 hybrid corns including 10 Korean silage corns developed by RDA such as Gangdaok, Kwangpyeongok, Dapyeongok, Andaok, Yanganok, Singwangok, Jangdaok, Cheongdaok, Pyeonggangok, and Pyeonganok as well as 40 foreign commercial silage corns. From cluster analysis, we confirmed that 10 Korean silage F1 hybrid corns were clearly distinguished except for Singwangok, P1395, and several foreign commercial corns, and selected minimum SNP primer combination for Gangdaok, Jangdaok, Pyeonggangok, and Pyeonganok. Therefore, development of SNP marker sets might be faster, cheaper, and feasible breed discrimination method through simple PCR and agarose gel electrophoresis.

Thermostable $\alpha$-Amylase Production by Thermophilic Bacillus sp. TR-25 lsolated from Extreme Enviroment (극한환경에서 분리한 고온성 Bacillus sp. TR-25에 위한 내열성 $\alpha$-amylase의 생산)

  • 노석범;손홍주;이종근
    • Journal of Life Science
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    • v.7 no.1
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    • pp.30-38
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    • 1997
  • For screening thermostable $\alpha$-amylase from thermophiles, various samples from extreme environments such as hot spring and sewage near them, and compoat, wereexamined microbial growth in enrichment culture medium at 55$\circ$C on the assumption that enzymes from thermophiles are inevitable thermostable. One strain showing higher $\alpha$-amylase activity was pure cultured and designated as Bacillus sp. TR-25 from the results of morphological, cultural and physiological characteristics. The most important carbon sourses for the enzyme production were soluble starch, dextrin, potato starch and corn starch. Glucose and fructose had a catabolite repression on the enzyme production. The good nitrogen sources for the enzyme production were yeat extract, nutrient broth, tryptone, corn steep liquor and ammonium sulfate. The enzyme production was accelerated by addition of CaCl$_{2}$. $\cdot $ H$_{2}$O. The optimal medium composition for the enzyme production was soluble starch 2.0%, yeast extract 0.55, CaCl$_{2}$ $\cdot $ 2H$_{2}$O 0.015, Tween 80 0.001%, pH8.0, respectively. In jar fermenter culture, this strain shows a rapid growth and required cheaper carbon and nitrogen source. These properties are very useful to fermentation industry. The $\alpha$-amylase of this strain demonstrated a maximum activity at 80$\circ$C, pH 5.0, respectively. And calcium ion did not improve thermostability of the enzyme. At 10$0^{\circ}C$, this enzyme has 235 of relative activity. Transformation was carried out by thermophilic Bacillus sp. TR-25 genomic DNA. As a result, the transformant has increased thermostable $\alpha$-amylase activity.

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Study on the Characteristics of DNA Comet Assay for Irradiated Vegetables and Grains (방사선조사된 채소류 및 곡물류의 DNA Comet Assay 특성 연구)

  • Seo, Jung-Eun;Oh, Se-Wook;Kim, Yun-Ji;Lee, Nam-Hyouck;Hong, Sang-Pill;Kim, Young-Ho
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.4
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    • pp.472-476
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    • 2008
  • The possibility of DNA comet assay as a rapid method for screening the irradiated vegetables and grains was evaluated. Vegetables (spring onion, garlic, and tomato) irradiated at $0{\sim}3$ kGy and grains (rice flour and black soybean) irradiated at $0{\sim}9$ kGy were used as samples. Optimum DNA comet assay conditions, such as elution, sedimentation of suspension, and lysis time of cell, were established. The optimum conditions for vegetables were 10 min for the elution time, 0 min for the sedimentation time, and 5 min for the lysis time. The optimum conditions for grains were 15 min for the elution time, 60 min for the sedimentation time, and 30 min for the lysis time. For the food application of DNA comet assay, it was possible to screen various food samples irradiated at the following doses: spring onion at 2 kGy, garlic at 3 kGy, tomato at 1 kGy, rice flour at 9 kGy, and black soybean at 3 kGy. Each sample showed different forms and sizes in DNA comet. Therefore, further studies on various methods using comet shape, concentration, or area in DNA comet assay are necessary.

Expression of Anthrax Lethal Factor, a Major Virulence Factor of Anthrax, in Saccharomyces cerevisiae (Yeast내에서 탄저병 원인균인 Bacillus anthracis의 치사독소인 Lethal Factor 단백질 발현)

  • Hwang Hyehyun;Kim Joungmok;Choi Kyoung-Jae;Chung Hoeil;Han Sung-Hwan;Koo Bon-Sung;Yoon Moon-Young
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.275-280
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    • 2005
  • Anthrax is an infectious disease caused by the gram-positive bacterium, Bacillus anthracis. Anthrax toxin is a tripartite toxin comprising of protective antigen (PA), lethal factor (LF) and edema factor (EF). PA is the receptor-binding component, which facilitates the entry of LF or EF onto the cytosol. LF is a zinc-dependent metalloprotease, which is a critical virulence factor in cytotoxicity of infected animals. Therefore, it is of interest to develop its potent inhibitors for the neutralization of anthrax toxin. The first step to identify the inhibitors is the development of a rapid, sensitive, and simple assay method with a high-throughput ability. Much efforts have been concentrated on the preparation of powerful assays and on the screening of inhibitors using these system. In the present study, we have tried to construct anthrax lethal factor in yeast expression system to prepare cell-based high-throughput assay system. Here, we have shown the results covering the construction of a new vector system, subcloning of LF gene, and the expression of target gene. Our results are first trial to express LF gene in eukaryote and provide the basic steps in design of cell-based assay system.

Analysis on the Relationships among the Total Cholesterol, Fasting Blood Sugar, Hypertension and Ischemic Heart Disease on EKG Findings (혈청 총콜레스테롤, 공복시 혈당, 혈압의 분포와 심전도상 허혈성 심질환과의 관련성)

  • Kim, Soo-Keun;Roh, Sang-Chul;Son, Jung-Il;Choi, Bo-Youl
    • Journal of Preventive Medicine and Public Health
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    • v.29 no.4 s.55
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    • pp.705-719
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    • 1996
  • Since the 1960s, Korea has achieved rapid economic growth, longer life expectancy, accelerated urbanization and a westenization of diet. Cardiovascular disease has become the leading cause of death; however the prevalence of ischemic heart disease(IHD) remains low. A cross-sectional multiphasic screening service for 18,426 persons aged $30\sim64$ years in a Medical Aid Program in Kyonggi-do Province was conducted from 1991 to 1993. Total cholesterol(TC), fasting blood glucose(FBG), blood pressure(BP), and electrocardiographic(EKG) data were collected. The result as follows ; 1. On the EKG findings, the age-adjusted prevalence of myocardial ischemia and myocardial infarction was 1.45% in men and 2.06% in women. 2. The mean blood pressure was 122.9/78.8mmHg. The age-adjusted prevalence of hypertension was 11.05% in men and 9.02% in women. The prevalence of hypertension showed increasing tendency according to age increase. In all age group, the prevalence of hypertension was higher in men than women. 3. The mean total cholesterol level was 184.4mg/dl in men and 189.2mg/dl in women. The age-adjusted prevalence of hypercholesterolemia was 4.88% in men and 5.67% in women. The total cholesterol level showed increasing tendency according to age increase, except $55\sim64$ age group in men. 4. The prevalence of hyperglycemia is 5.8%. The age-adjusted prevalence of hyperglycemia is 6.72% in men and 4.50% in women. The prevalence of hyperglycemia showed increasing tendency according to age increase. 5. On the EKG findings, the prevalence of myocardial ischemia and myocardial infarction was higher in hypertension than normal, in all age group of men and women less than 40 years-old. Only in women more t]fan 40 years-old, the prevalence of myocardial ischemia and myocardial infarction was higher in hypercholesterolemia and hyperglycemia. Nevertheless there is not statistical association between ischemia heart disease and previous risk factors in other age group, the prevalence of myocardial ischemia and myocardial infarction was higher in hypertension, hypercholesterolemia and hyperglycemia than normal. The result of this study suggest that relationships between major risk factor of ischemic heart disease and ischemic heart disease is similar to existing theory.

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Effective Screening Methods for Lipoxygenase Isozymes in Soybean Seeds (콩 lipoxygenase 효소의 효율적인 검정법)

  • Kim, Young Jin;Park, Tae Il;Cho, Sang Kyun;Oh, Young Jin;Kim, Tae Soo;Kim, Jung Gon
    • Korean Journal of Breeding Science
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    • v.40 no.1
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    • pp.26-30
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    • 2008
  • Normal soybean seed contains three lipoxygenase isozymes called L-1, L-2, and L-3, respectively, which are responsible for the generation of undesirable grassy-beany flavors. Simple and effective methods for the detection of lipoxygenase isozymes were developed in soybean seeds. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) has been tried in separating these isozymes. It was done effectively on 7.5% separating gel and 4.5% stacking gel. However, no reliable method has been developed specifically for separating L-3, L-13 and L-23. Visual judging methods were based on the bleaching activities of lipoxygenase in contact with methylene blue and ${\beta}$-carotene. Sodium linoleate bleaching method was adopted to determine L-1 and L-2. Carotene bleaching and spectrophotometric methods were used to determine L-3. These systems were very rapid within one minute, furthermore only required a small piece of cotyledon (below 10 mg) and the other part could be used for generation advance after analysis. It was demonstrated that 200 seed samples could be analyzed per day by one laboratory assistant. The combination of visual judging methods and electrophoresis is suitable for breeding programs. It took 6.5 hours for analysis of 100 seed samples by one person.

Investigation of the Molecular Diagnostic Market in Animals (동물 분자 진단 시장의 동향)

  • Park, Chang-Eun;Park, Sung-Ha
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.1
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    • pp.26-33
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    • 2019
  • Recently, the rapid growth of the companion animal market has led to the development of animal disease diagnosis kits. Therefore, the utility of the introduction of biomarkers for the development of animal molecular diagnostics is being reevaluated. A good biomarker should be precise and reliable, distinguish between normal and diseased states, and differentiate between different diseases. Recently reported genetic markers, tumor markers (cell free DNA, circulating tumor cells, granzyme, and skin tumors), and others (brucellosis, programmed death recovery-1, symmetric dimethylarginine, periostin, and cysteinyl leukotrien) have been developed. The biomarkers are used for risk prediction or for the screening, diagnosis, and monitoring of disease progression. The most important criteria for related biomarkers are disease specificity. Many potential biomarkers have emerged from laboratory and test studies, but they have not been validated in independent or large-scale clinical studies. Candidate biomarkers evaluate disease associations, verify the effectiveness of biomarkers for early detection and disease progression, and incorporate them into humans and animals. In the future, it will be necessary to reevaluate the utility of well-structured biomarker-based research and study the development of kits that can be used in on-site tests in accordance with the trends introduced in the diagnosis of animal diseases.

Establishment of a Nondestructive Analysis Method for Lignan Content in Sesame using Near Infrared Reflectance Spectroscopy (근적외선분광(NIRS)을 이용한 참깨의 lignan 함량 비파괴 분석 방법 확립)

  • Lee, Jeongeun;Kim, Sung-Up;Lee, Myoung-Hee;Kim, Jung-In;Oh, Eun-Young;Kim, Sang-Woo;Kim, MinYoung;Park, Jae-Eun;Cho, Kwang-Soo;Oh, Ki-Won
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.67 no.1
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    • pp.61-66
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    • 2022
  • Sesamin and sesamolin are major lignan components with a wide range of potential biological activities of sesame seeds. Near infrared reflectance spectroscopy (NIRS) is a rapid and non-destructive analysis method widely used for the quantitative determination of major components in many agricultural products. This study was conducted to develop a screening method to determine the lignan contents for sesame breeding. Sesamin and sesamolin contents of 482 sesame samples ranged from 0.03-14.40 mg/g and 0.10-3.79 mg/g with an average of 4.93 mg/g and 1.74 mg/g, respectively. Each sample was scanned using NIRS and calculated for the calibration and validation equations. The optimal performance calibration model was obtained from the original spectra using partial least squares (PLS). The coefficient of determination in calibration (R2) and standard error of calibration (SEC) were 0.963 and 0.861 for sesamin and 0.875 and 0.292 for sesamolin, respectively. Cross-validation results of the NIRS equation showed an R2 of 0.889 in the prediction for sesamin and 0.781 for sesamolin and a standard error of cross-validation (SECV) of 1.163 for sesamin and 0.417 for sesamolin. The results showed that the NIRS equation for sesamin and sesamolin could be effective in selecting high lignan sesame lines in early generations of sesame breeding.