• Title/Summary/Keyword: Biological screening

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Anti-inflammatory and Anti-oxidative Activities of Flavonoids Extracted from Dendranthema indicum Flowers in Jeju Island (제주 자생 감국 꽃 추출물 유래 flavonoid 화합물의 항산화 및 항염 활성)

  • Hyun, Ju Mi;Jo, Yeon Jeong;Kim, Yun Beom;Park, Sung-Min;Yoon, Kyung-Sup;Lee, Nam Ho
    • Journal of the Korean Applied Science and Technology
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    • v.36 no.4
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    • pp.1259-1267
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    • 2019
  • Anti-inflammatory and anti-oxidative activities were examined on the extract of Dendranthema indicum (D. indicum) flowers. The flowers were extracted two times for 24 h each with 70% ethanol. Upon the biological activities screening, the ethanol extract exhibited potent free radical scavenging activities and inhibited the production of nitric oxide on LPS-induced RAW264.7 macrophages effectively without causing cell toxicity. Further purification by medium pressure liquid hromatography (MPLC) and identification of the isolates led to identification of cynaroside (1) and apigetrin (2). The chemical structures of the isolated compounds were elucidated based on spectroscopic data including nuclear magnetic resonance (NMR) spectra, as well as comparison of the data to the literature values. Also, the quantitative analysis of the compounds was perfromed by high-performance liquid chromatography (HPLC). The isolates 1 and 2 were determined to inhibite the nitric oxide (NO) production dose-dependently. Based on these results, it was suggested that D. indicum extract could be useful as anti-inflammatory agents in cosmetics applications.

Isolation and Characterization of Zymomonas mobilis DNA Fragments Showing Promoter Activity in Escherichia coli (Escherichia coli에서 Promoter 활성을 보이는 Zymomonas mobilis DNA 조각의 분리와 분석)

  • Kim, Eun-Joon;Yoon, Ki-Hong;M.Y. Pack
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.600-605
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    • 1989
  • For the purpose of isolation of the Zymomonas mobilis DNA fragments showing promoter activity in Escherichia coli, a promoter screening vector, PCMT215 was constructed by transferring a promoterless chloramphenicol acetyltransferase (CAT) gene of pYEJ001 into pMT21 which contains $\beta$-lactamase gene and multiple cloning sites. A library of Z, mobilis Sau3AI DNA fragments was constructed in E. coli using the newly constructed pCMT215. Fourteen clones showing resistance to chloramphenicol ranging in concentration from 30 to 750 $\mu$g/$m\ell$ were selected. From five clones of them, the Z. mobilis DNA fragments expressing CAT gene of the recombinant plasmids were sequenced and then sites of transcriptional initiation were identified. The nucleotide sequences of the cloned DNA shared AT rich regions, poly A's or T's stretches and palindromic regions. The positions of transcriptional initiation for CAT gene occurred at more than one site spaced over by 4 to 190 base pairs on the cloned fragments in E. coli.

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Screening of the Genes Expressed in Pichia pastoris Grown in Phosphate-Limited Chemostat Culture (인산제한상태에서 발현되는 Pichia pastoris 유래 유전자 탐색)

  • Hong, Ji-Yeon;Ahn, Jung-Oh;Park, Myoung-Soo;Choi, Soon-Yong;Choi, Eui-Sung;Jung, Joon-Ki;Lee, Hong-Weon
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.272-277
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    • 2007
  • The physiological responses of microorganisms to specific nutrient limitation can be regulated at the transcriptional levels. In this study, in order to develop the Pichia pastoris-derived promoter inducible by nutrient-limited condition, we constructed cDNA libraries using RT-PCR of total RNA from P. pastoris in steady-states of phosphate-limited chemostat with different dilution rates. Various genes were detected from cDNA library. Among these genes, the gene encoding putative sodium/phosphate ($Na^+$/Pi) symporter (NPS), high affinity transporter of phosphate, was detected. It was observed that expression of NPS increased in a manner specific to phosphate-limited condition through Northern blot. Therefore, it is thought that the promoter from NPS gene may have the potential as auto-inducible promoter by phosphate-limited culture condition without inducer.

Production of Monoclonal Antibody Against Listeria monocytogenes and Its Application to Immunochromatography Strip Test

  • Shim, Won-Bo;Choi, Jin-Gil;Kim, Ji-Young;Yang, Zheng-You;Lee, Kyu-Ho;Kim, Min-Gon;Ha, Sang-Do;Kim, Keun-Sung;Kim, Kwang-Yup;Kim, Cheol-Ho;Ha, Kwang-Soo;Eremin, Sergei A.;Chung, Duck-Hwa
    • Journal of Microbiology and Biotechnology
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    • v.17 no.7
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    • pp.1152-1161
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    • 2007
  • An immunochromatography (ICG) strip test based on a monoclonal antibody for the rapid detection of L. monocytogenes in meat and processed-meat samples was developed in this study. A monoclonal antibody (MAb) specific to L. monocytogenes was produced from cloned hybridoma cells (FKLM-3B12-37) and used to develop an ICG strip test. The antibody showed a stronger binding to L. monocytogenes than other Listeria species, and a weak cross-reaction to S. aureus based on an ELISA. The detection limit of the ICG strip test was $10^5\;cell/ml$. In total, 116 meat and processed-meat samples were collected and analyzed using both the ICG strip test and a PCR. The ICG strip test and PCR indicated L. monocytogenes contamination in 34 and 27 meat samples, respectively. The 7 meat samples not identified as L. monocytogenes positive by the PCR were also tested using an API kit and found to be contaminated by Listeria species. In conclusion, the ICG strip test results agreed well with those obtained using the PCR and API kit. Thus, the developed ICG has potential use as a primary screening tool for L. monocytogenes in various foods and agricultural products, generating results within 20 min without complicated steps.

Screening Test(I) of Several Antipsychotic Agents on NO Formation (수종 정신병치료제들의 NO형성에 대한 검색(I))

  • Lee, Jong-Hwa;EI-fakahany, Esam E.
    • The Korean Journal of Pharmacology
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    • v.30 no.3
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    • pp.343-349
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    • 1994
  • A number of neurological syndromes(e.g. tardive dyskinesia) are developed as a consequence of chronic treatment with neuroleptics or antipsychotic agents. Despite the long and succesful use of phenothiazine derivatives and related agents in the treatment of certain states of mental disease, the mechanisms of these drugs are still poorly understood. One current hypothesis from extensive reviews is that these compounds might significantly interfere with the cyclic nucleotide system in brain (Levin and Weiss, 1977; Nowicki et al., 1991; Haley et al., 1992). Nitric oxide (NO), one of an interesting messenger molecule and aberrant transmitter, is believed to play a important role in biological functions of cyclic nucleotides in nervous system. It has been reported that calcium-dependent NO synthesis in endothelial cytosol is mediated by calmodulin which is supposed to be tightly related to pharmacological actions of antipsychotic agents. In the present study, the effect of several antipsychotic agents on the activity of NO synthesis and formation of cyclic GMP were investigated. These agents inhibited both the formation of $[^3H]L-citrulline$ and that of $[^3H]cyclic$ GMP by concentration-dependent manner, and their inhibiting patterns are so similar to that of calmodulin antagonist.

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Feature Analysis of Different In Vitro Antioxidant Capacity Assays and Their Application to Fruit and Vegetable Samples (In Vitro 항산화능 측정법에 대한 특징 분석과 채소.과일 시료에 대한 적용 사례 고찰)

  • Kim, Min-Jung;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.7
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    • pp.1053-1062
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    • 2011
  • Reactive oxygen species (ROS), including singlet oxygen (${O_2}^1$), superoxide anion radical ($O_2{\cdot}^-$), hydroxyl radical ($HO{\cdot}$), peroxyl radical ($ROO{\cdot}$), hydrogen peroxide ($H_2O_2$), and hypochlorous (HOCl), are generated as byproducts of normal cellular metabolism. ROS induce damage to many biological molecules, such as lipids, proteins, carbohydrates, and DNA. It is widely believed that some degenerative diseases caused by ROS can be prevented by the high intake of fruits and vegetables due to their antioxidant activities. Recently, research on natural antioxidants has become increasingly active in various fields. Several assays have been developed to measure the total antioxidant capacity of antioxidants in fruits and vegetables in vitro. These assays include those for DPPH radical scavenging activity, SOD-like activity, total polyphenol content, oxygen radical absorbance capacity, reducing power, trolox equivalent antioxidant capacity (ABTS assay), single-cell gel electrophoresis (comet assay), and a cellular antioxidant activity assay. Because different antioxidant compounds may act through different mechanisms in vitro, no single assay can fully evaluate the total antioxidant capacity of foods. Due to the complexity of the composition of foods, it is important to be able to measure antioxidant activity using biologically relevant assays. In this review, recently used assays were selected for extended discussion, including a comparison of the advantages and disadvantages of each assay and their application to fruits and vegetables.

The Role of Serum Pepsinogen and Gastrin Test for the Detection of Gastric Cancer in Korea (한국인 위암 진단에 있어 혈청 펩시노겐과 혈청 가스트린 검사의 역할)

  • Kim, Na-Young
    • Journal of Gastric Cancer
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    • v.9 no.3
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    • pp.78-87
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    • 2009
  • Serum pepsinogen (sPG) is a marker of gastric mucosal atrophy, a condition that has been associated with an increased risk of gastric neoplasia. A low sPGI level and a low PG I/II ratio have been associated with severe gastric atrophy, and are frequently found in gastric cancer. Because the prevalence of gastric cancer is high in Korea, it would be convenient if a good biomarker for gastric cancer were developed. Two studies recently investigated the efficacy of sPG along with Helicobacter pylori (H. pylori) as a screening tool for gastric cancer. In these studies, sPG was measured using a Latex enhanced Turbidimetric Immunoassay. We found that H. pylori IgG status, age and gender were associated with serum pepsinogen levels. Thus, to increase the ability of the PG I/II ratio to detect atrophic gastritis, the cutoff value for the PG I/II ratio should be stratified according to the H. pylori IgG status. In addition, a PG I/II ratio ($\leq3.0$), which has been widely used as an international standard for gastric cancer, was found to be a reliable marker for the detection of gastric dysplasia or gastric cancer, especially of the intestinal type. The efficacy of the test in Korea was lower than the efficacy in Japan. However, the detecting power of a PG I/II ratio ($\leq3.0$) was significantly increased in the presence of H. pylori. The ratio together with H. pylori psotivitiy could provide a means of identifying persons at high risk of developing gastric cancer in Korea.

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Eleocharis kuroguwai Ohwi Ameliorates LPS-mediated Inflammation by Suppressing MAPKs Signaling (LPS로 유도된 염증 상태 내에서 MAPKs 세포신호 전달체계를 저해하는 올방개의 항염증 효과)

  • Kim, Kang-Hoon;Bae, Eun Young;Lim, Hyun Goo;Kim, Mi Heui;Oh, Tae-Seok;Lee, Seung Woong
    • Journal of Life Science
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    • v.30 no.5
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    • pp.476-482
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    • 2020
  • In rice agriculture, Eleocharis kuroguwai Ohwi (olbanggae) is a target for herbicidal intervention as a problem weed although it has also long been used clinically as a traditional medicine for jaundice, fever, and blood flow. E. kuroguwai has been evaluated in many clinical trials, but its molecular biological advantages are still unknown. Here, we investigate the anti-inflammatory effects of E. kuroguwai 80% ethanol extracts by screening NO production in LPS-induced macrophage activation. To find the most effective fractions, we partitioned five sub-fractions using HP20 column chromatography, namely 20%, 40%, 60%, 80%, and 100%. Of these, the 60% and 80% sub-fractions were found to significantly inhibit NO production; there were no toxicological effects at any concentration. In addition, the 80% sub-fraction inhibited significantly the iNOS and the mRNA of the pro-inflammatory mediators IL-6, TNF-α, and IL-1β by inhibiting the phosphorylation of JNK and ERK pathways associated with MAPKs signaling. Our results suggest that the 80% E. kuroguwai sub-fraction has the most significant anti-inflammatory effects of inhibiting iNOS and pro-inflammatory mediators and suppressing the phosphorylation of JNK and ERK. Therefore, the 80% sub-fraction of E. kuroguwai extract may be a therapeutic candidate for inflammatory diseases associated with the overexpression of MAPKs.

Screening of Nine Herbs with Biological Activities on ACE Inhibition, HMG-CoA Reductase Inhibition, and Fibrinolysis (9종의 허브류로부터 ACE 저해활성, HMG-CoA reductase 저해활성 및 혈전용해활성에 대한 검색)

  • Kwon, Eun-Kyung;Kim, Young-Eon;Lee, Chang-Ho;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.38 no.5
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    • pp.691-698
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    • 2006
  • The purpose of this study was to investigate how herb extracts may improve blood circulation. Twenty-six extracts from nine different herbs (marjoram, lavender, dill, rosemary, hyssop, rose, lemon balm, pineapple sage, and echinacea) were evaluated for their anti-hypertensive effects via angiotensin I converting enzyme (ACE) inhibition. Their cholesterol-lowering effects via hydroxy-methyl-glutaryl coenzyme A (HMG-CoA) reductase inhibition and their fibrinolytic activity via fibrin-plate method were also evaluated. Both water extraction of rose flowers and 70% EtOH extraction of pineapple sage leaves effectively reduced the ACE activity with inhibition rates of 133.8% and 91.2%, respectively. Similarly, both water and 70% EtOH extracts of rose flowers strongly inhibited the enzymatic activity of HMG-CoA reductase by 48.9% and 80.5%, respectively. Water and 70% EtOH extracts of rose flowers also showed relatively high fibrinolytic activity. Based on these observations, rose flower extracts can be developed as a functional tool for use in the improvement of blood circulation.

The Effects of Cesium, Strontium and Cobalt on Cell Toxicity in the 2D and 3D Cell Culture Platforms (단층 및 입체 세포배양환경에서 세슘, 스트론튬 및 코발트가 세포 독성에 미치는 영향 분석)

  • Kim, Gi Yong;Kang, Sung-Min;Jang, Sung-Chan;Huh, Yun Suk;Roh, Changhyun
    • Korean Journal of Environmental Biology
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    • v.34 no.2
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    • pp.107-115
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    • 2016
  • Currently, there are 442 operating nuclear power plants in the world, and 62 more are under construction. According to this reasoning, the treatment of radioactive waste is important to prevent the environmental ecosystem including humans, animals, and plants. Especially, a leakage of radioactive waste causes not only regional problem but also serious global one. In this study, we demonstrate the effect of radioisotopes (e.g., cesium, strontium, and cobalt) on a 3D culture cell. To develop the 3D cell culture system, we used a 96-well-culture plate with biocompatible agarose hydrogel. Using this method, we can perform the 3D cell culture system with three different cell lines such as HeLa, HepG2, and COS-7. In addition, we conducted a cell viability test in the presence of radioisotopes. Interestingly, the 3D morphological cells showed 42% higher cell viability than those on the 2D against cesium. This result indicates that the 3D platform provides cells morphological and physiological characteristic similar to in vivo grown tissues. Moreover, it overcomes the limitation of conventional cell culture system that can't reflect in vivo systems. Finally, we believe that the proposed approach can be applied a new strategy for simple high-throughput screening and accurate evaluation of metal toxicity assay.