• 제목/요약/키워드: Indicator assay

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Soil Pollution Assessment Based on Ecotoxicological Methods (생태독성학적 기법을 이용한 토양오염평가 방안)

  • An Youn-Joo;Jeong Seung-Woo
    • Journal of Soil and Groundwater Environment
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    • v.10 no.6
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    • pp.56-62
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    • 2005
  • Chemical analyses are generally used to assess contaminated soils and to monitor the efficiency of soil remediation. In this study, the ecotoxicological methods was suggested to evaluate soil pollution by using a battery of bioassay. Plant assay and earthworm assay were conducted to evaluate ecotoxicity o soils contaminated by heavy metals (cadmium and copper) and oil (BTEX compounds, toluene). Test plants were Zea may, Triticum aestivum, Cucumis sativus, and Sorghum bicolor. The presence of heavy metals decreased the seedling growth. Cucumis sativus and Sorghum bicolor seemed to be good indicator plants which are sensitive to heavy metal pollution as well as BTEX contamination. An earthworm bioassay was performed to predict the ecotoxicity in toluene-contaminated soils, based on a simple contact method. Perionyx excavatus was adopted as a test earthworm species, and the severity of response increased with increasing toluene concentration. The present study demonstrated that ecotoxicological methods could be a quantitative approach to evaluate contaminated soils.

Characterization of Antimicrobial Substance Produced by Lactobacillus paraplantarum KNUC25 Isolated from Kimchi (김치로부터 분리된 Lactobacillus paraplantarum KNUC25가 만드는 항균 물질의 특성)

  • Kim, Ma-Rie;Lee, Su-Jin;Seul, Keyung-Jo;Park, Yu-Mi;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.24-32
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    • 2009
  • The KNUC25 strain isolated from over-fermented whole Chinese cabbage kimchi was examined for its physiological characteristics using API 50 CHL system assay and identified as Lactobacillus paraplantarum by analysis of whole-cell protein SDS-PAGE pattern assay and similarity of 16S rDNA sequence. L. paraplantarum KNUC25 had a broad antimicrobial activity spectrum from Gram positive to Gram negative bacteria. Scanning electron micrograph analysis showed that KNUC25 might attack to cell surface of indicator cells and destruction can lead to inhibition of the cell growth. The antimicrobial substance of the KNUC25 strain was stable to various degrading enzymes and at high temperature and not a plasmid-born matter. Resistance to proteolytic enzymes showed that an antimicrobial activity of KNUC25 might not be caused by proteinous substance. Maximum production of antimicrobial substance was the exponential growth phase at $30^{\circ}C$.

Increased Frequency of Micronuclei in Binucleated Lymphocytes among Occupationally Pesticide-exposed Populations: A Meta-analysis

  • Yang, Hai-Yan;Feng, Ruo;Liu, Jing;Wang, Hai-Yu;Wang, Ya-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.16
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    • pp.6955-6960
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    • 2014
  • Background: The cytokinesis-block micronucleus (CBMN) assay is a standard cytogenetic tool employed to evaluate chromosomal damage subsequent to pesticide exposure. Objectives: To evaluate the pooled levels of total micronuclei (MN) and binucleated cells with micronuclei (MNC) in 1000 binucleated lymphocytes among population occupationally exposed to pesticides and further determine the more sensitive biomarker of CBMN. Materials and Methods: A meta-analysis on the pooled levels of MN and MNC in binucleated lymphocytes among occupationally pesticide-exposed populations was conducted using STATA 10.0 software and Review Manager 5.0.24 in this study. Results: We found significant differences in frequencies of MN and MNC in 1000 binucleated lymphocytes between pesticide-exposed groups and controls, and the summary estimates of weighted mean difference were 6.82 [95% confidence interval (95% CI): 4.86-8.78] and 5.08 (95% CI: 2.93-7.23), respectively. However, when we conducted sensitivity analyses further, only the MN remained statistically different, but not the MNC, the summary estimates of weight mean difference were 2.86 (95% CI: 2.51-3.21) and 0.50 (95% CI: -0.16-1.17), respectively. We also observed pesticide-exposed subjects had significantly higher MN frequencies than controls among smokers and nonsmokers, male and female populations, and American, Asian and European countries in stratified analyses. Conclusions: The frequency of MN in peripheral blood lymphocytes might be a more sensitive indicator of early genetic effects than MNC using the CBMN assay for occupationally pesticide-exposed populations.

Antioxidant, antinociceptive activity and general toxicity study of Dendrophthoe falcata and isolation of quercitrin as the major component

  • Hasan, Md Shihab;Ahmed, Md Iqbal;Mondal, Sukla;Masud, Mohammad Methedi;Sadhu, Samir Kumar;Ishibashi, Masami;Uddin, Shaikh Jamal
    • Advances in Traditional Medicine
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    • v.6 no.4
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    • pp.355-360
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    • 2006
  • Plants have been used in traditional medicinal system for centuries. Bangladeshi medicinal plants have received considerable attention from the researchers for evaluation of their bioactivity. As a part of our ongoing research of screening the Bangladeshi medicinal plants, the ethanolic extract of Dendrophthoe falcata have been chosen for the present study. The ethanolic extract of the leaves of the plant have been assessed for their antioxidant, antinociceptive, and general toxicity. The extract showed potent antioxidant activity ($IC_{50}5.1{\mu}g/ml$) using DPPH radical scavenging assay, which is comparable to the standard ascorbic acid ($IC_{50}4.6{\mu}g/ml$). The extract significantly and dose dependently inhibited the acetic acid induced writhing in mice (71.2%, P < 0.001 and 28.0%, P < 0.05 for 500 and 250 mg/kg body weight, respectively). A general toxicity was assessed by a simple and low cost assay using brine shrimp lethality as an indicator. The extract showed low level of toxicity ($LC_{50}100{\mu}g/ml$). Using different chromatographic techniques, quercitrin (quercetin 3-O-${\alpha}$-rhamnoside) was separated as the major component from the extract. The structure was elucidated by detailed 1D and 2D NMR and mass spectral analysis.

Quality Changes of Fresh Vegetable and Fruit Juice by Various Juicers (주스제조 장치에 따른 채소 및 과일 주스의 품질 변화)

  • Choi, Moon-Hee;Kim, Min-Joo;Jeon, Young-Jin;Shin, Hyun-Jae
    • KSBB Journal
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    • v.29 no.3
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    • pp.145-154
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    • 2014
  • A fresh vegetable and fruit juice has become a new healthy food available for detoxification, dieting and health. This paper presents the useful information about the quality changes of fresh juice according to different juicer. Quality of fresh juice could be evaluated by several factors such as juice yield, enzyme activity, antioxidant activity, polyphenol contents, and anti-inflammatory activity. The juice yields of 12 different vegetables and fruits were compared using 6 different juicers and it was observed that the yield of slow juicer was better than that of conventional blender. Among 12 samples, the juice yield of grape is the best and the pH of the juice was in the acidic range of 3 and 4. Kiwi and grapefruit were the best in terms of protease enzyme activities by Hemoglobin units on the tyrosine basis and Spectrophotometric acid protease unit and papain units on the tyrosine basis of KFDA protocols. The total polyphenol contents were also high in kiwi and grapefruit. The antioxidant activity by diphenyl-1-picrylhydrazyl (DPPH), superoxide dismutase (SOD), and radical scavenging assay were high in the order of kiwi, grapefruit, grape, tomato, and orange. Anti-inflammatory activities were also assay for 12 samples with 6 juicers. It can be concluded that of fresh fruit and vegetable juice provides a source of antioxidant components and enzymes with high activity. And the enzyme activities could be used as one of the quality indicator of fresh juice. Concerning the juicers used in this study, slow juicer could be recommended to prepare the fresh juice in terms of the juice quality.

DNA Repair Capacity in Peripheral Blood Lymphocytes Predicts Efficacy of Platinum-based Chemotherapy in Patients with Gastric Cancer

  • Zhang, Yi-Yin;Gu, Kang-Sheng
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5507-5512
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    • 2013
  • Objective: To investigate the correlation between ERCC1 expression levels in tumor tissue and peripheral blood lymphocytes (PBL) from patients with gastric cancer and assess the relationship between PBL DNA repair rate (DRR) and the efficacy of platinum chemotherapy. Methods: A total of 53 patients with gastric cancer receiving surgery and 20 controls were studied. ERCC1 protein expression in tumour tissue and PBL were determined by immunohistochemical staining. The PBL DRRs of 47 advanced patients and 20 controls were estimated by comet assay. Results: The positive expression rates of ERCC1 were 67. 9%, 56. 6% and 10.0% in tumour tissues, PBLs of gastric cancer patients, and PBLs of the control group. PBL ERCC1 expression correlated with that in tissue (${\chi}^2$=15. 463, p=0.000). Pearson contingency coefficient=0.475). DRRs of cancer patients by tail length (TL) (Z=4. 662, p=0.000) and tail moment (TM) (Z=3. 827, p=0.000) were significantly lower than that of control group. When TL was applied as an indicator, the correlation between DRR and chemotherapy efficacy was significant (Spearman rank correlation r=0.327, p=0.032). Patients with low levels of DRR in PBL presented better short-term efficacy of chemotherapy than those with high levels of DRR. Conclusions: The ERCC1 expression in PBLs may indirectly reflect ERCC1 expression in gastric cancer tissues. Compared with non-cancer populations, patients with gastric cancer may have lower DNA repair capacity. DRR in PBL may predict the short-term efficacy of platinum-based chemotherapy for patients with advanced gastric cancer.

Zinc Status Assessment by Analysis of Mononuclear Cell Metallothionein mRNA Using Competitive-Reverse Transcriptase-Polymerase Chain Reaction

  • Lee, Soo-Lim;Yoon, Jin-Sook;Kwon, Chong-Suk;Beattie, John H.;Kwun, In-Sook
    • Preventive Nutrition and Food Science
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    • v.9 no.3
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    • pp.276-282
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    • 2004
  • Marginal Zn deficiency is prevalent through the world and yet human zinc status has not been properly assessed due to the lack of a reliable diagnostic indicator. One potential possibility for zinc status assessment using Zn-binding protein, metallothionein (MT)-mRNA, has been proposed. The purpose of the present study was aimed to show whether measurement of mononuclear cell (MNC) MT mRNA, using a competitive-reverse transcriptase-polymerase chain reaction (competitive-RT-PCR) assay, could indicate zinc status in human subjects. In this study, MNC MT-mRNA expression was measured using a competitive-RT-PCR to compare before and after 14 days of zinc supplementation (50 mg Zn/das zinc gluconate). RT-PCR oligonucleotide primers which were designed to amplify both a 278 bp segment of the human MT-2A cDNA and a 198 bp mutant competitor cDNA template from MNCs, were prepared. MT-2A mRNA was normalized by reference to the housekeeping gene, $\beta$-actin, mRNA for which was also measured by competitive-RT-PCR. There was considerable inter-individual variation in MT-mRNA concentration and yet, the mean MT-2A mRNA level increased 4.7-fold after Zn supplementation, as compared to before Zn supplementation. This MT-2A mRNA level was shown as the same pattern and, even more sensitive assay, compared to the conventional plasma and red blood cells (RBCs) Zn assessment in which plasma and RBCs zinc levels increased 2.3- and 1.2-fold, respectively (p<0.05). We suggest that MT competitive-RT-PCR can be a useful assessment tool for evaluating human zinc status.

VALIDATION AND UTILIZATION OF THE SKINTEXTM SYSTEM

  • Gordon, V.C.;Realica, B.;Tolstrup, K.;Puls, B.
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.17 no.1
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    • pp.64-80
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    • 1991
  • The SKINTEX Method is based on a two-compartment physico-chemical model which includes a Biomembrane Barrier in compartment one and an organized macromolecular matrix in compartment two. Test samples absorb onto or permeate through the keratin/collagen Biomembrane Barrier and then can interact with the organized macromolecular matrix. Changes in the integrity of the barrier release a dye indicator: Changes in the matrix can alter its transparency. The sum of these two responses is read spectrophotometrically at 470nm. An early investigation of 950 chemicals and formulations in the SKINTEX System produced results which were 89% concordance to in vivo Draize dermal irritation results obtained with 24-hour occluded application of test samples with-out abrasion and standard scoring. Alkaline materials were analyzed in a specialized SKINTEX AMA Protocol. In this early study, the model did not distinguish nonirritant test materials and formulation with PDII(Primary Dermal Irritation Index)in the range from 0 to 1.2, A High Sensitivity Assay Protocol(HSA)was developed to amplify the changes in both compartments of this model and provide more accurate calibration of these changes. A study of 60 low irritation test samples including cosmetics, household products, chemicals and petro-chemicals distinguished nonirritants with PDII $\leq$ 0.7 for 26 of 30 nonirritants. A second protocol was developed to evaluate the SKINTEX model predictability with respect to human irritation. The Human Response Assay (HRA )has been optimized based on differences in penetration and irritation responses in humans and rabbits. An additional 32 test materials with different mechanisms and degrees of dermal toxicity were evaluated by the HRA. These in vitro results were 86% concordant to human patch test results. In order to further evaluate this model, a Standard Chemical Labelling (SCL) Protocol was developed to optimize this system to predict Draize dermal irritation results after a 4-hour application of the test material. In a study of 52 chemicals including acids, bases, solvents, salts, surfactants and preservatives, the SCL results demonstrated 85% concordance to Draize results for a 4-hour application of test samples on non-abraded rabbit skin. The SKINTEX System, including three specialized protocols, provided results which demonstrated good correlation to the endpoint of dermal irritation in man and rabbits at different application times.

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Genetic and Epigenetic Biomarkers on the Personalized Nutrition

  • An Sung-Whan
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
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    • 2004.11a
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    • pp.271-274
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    • 2004
  • Nutritional genomics is a new field of study of how nutrition interacts with an individual's genome or individual responds to individual diets. Systematic approach of nutritional genomics will likely provide important clues about responders and non-responders. The current interest in personalizing health stems from the breakthroughs emerging in integrative technologies of genomics and epigenomics and the identification of genetic and epigentic diversity in individual's genetic make-up that are associated with variations in many aspects of health, including diet-related diseases. Microarray is a powerful screen system that is being also currently employed in nutritional research. Monitoring of gene expression at genome level is now possible with this technology, which allows the simultaneous assessment of the transcription of tens of thousands of genes and of their relative expression of pathological cells such tumor cells compared with that of normal cells. Epigenetic events such as DNA methylation can result in change of gene expression without involving changes in gene sequence. Recent developed technology of DNAarray-based methylation assay will facilitate wide study of epigenetic process in nutrigenomics. Some of the areas that would benefitfrom these technologies include identifying molecular targets (Biomarkers) for the risk and benefit assessment. These characterized biomarkers can reflect expose, response, and susceptibility to foods and their components. Furthermore the identified new biomarker perhaps can be utilized as a indicator of delivery system fur optimizing health.

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Development of an Agar Diffusion Method to Measure Elastase Inhibition Activity Using Elastin-Congo Red

  • Jung Kyung-Hwan;Kim Hyun-Joo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1320-1324
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    • 2006
  • The pancreatic and neutrophil elastases are associated with several illnesses including lung and vascular diseases, various cancers, and pancreatitis. The development of a potent and specific inhibitor to the elastases could lead to new therapies. In this study, an agar diffusion method was modified to include a substrate-dye conjugate (Elastin-Congo red) as a substrate of elastase and an indicator of elastase inhibitory activity. The Elastin-Congo red agar plates consisted of 0.1 % Elastin-Congo red and 2.5% agar. The elastase and elastase inhibitors were simultaneously loaded into wells, ultimately resulting in halo formations in which the halo diameter decreased as the concentration of elastase inhibitor increased. The concentration of elastase inhibitor in the samples, therefore, was inversely proportional to the halo diameters. This simplified method provided an excellent correlation with the standard microplate technique, which uses a chromogenic substrate. The concentration of elastase inhibitor obtained from the culture supernatant of a recombinant elastase inhibitor produced by the yeast Pichia pastoris was easily determined. This study has established a simple modified and inexpensive agar diffusion method that is potentially useful for the identification, quantification, and screening of new elastase inhibitors.