• Title/Summary/Keyword: Binding Potential

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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.

Anti-Obese Activity of HPJ Extract on High Fat Diet-Induced Obese Mice (고지방 식이로 유도된 비만 쥐에서 HPJ 추출물의 항비만 효과)

  • Yuan, Hai-Dan;Quan, Hai-Yan;Zhang, Ya;Kim, Sung-Jib;Shin, Dae-Hee;Lim, Bang-Ho;Chung, Sung-Hyun
    • YAKHAK HOEJI
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    • v.53 no.5
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    • pp.286-292
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    • 2009
  • In this study, we investigated the anti-obese activity of HPJ extract in C57BL/6J mice. The C57BL/6J mice were randomly divided into five groups: normal control group (Con), high fat diet control group (HFD), treatment groups with HPJ at 125 mg/kg (HPJ125), 250 mg/kg (HPJ250), or 500 mg/kg (HPJ500). To induce an obesity, mice were fed by a high fat diet for 6 weeks, and mice were administered with HPJ extract once a day for 8 weeks. At the end of treatment, we examined the effect of HPJ extract on body weight, plasma lipid, and lipogenic enzymes. HPJ extract was found to lower whole body and epididymal adipose tissue weights and lowered plasma levels of glucose, insulin, triglyceride (TG), total cholesterol (TC), non-esterified fatty acid (NEFA) and leptin, compared to those in HFD group. Histological analyses of the liver and fat tissues of mice treated with HPJ extract revealed significantly decreased number of lipid droplets and decreased size of adipocytes compared to the HFD group. In addition, HPJ extract preserved the morphological integrity of pancreatic islets. To elucidate an action mechanism of HPJ extract, Western blot and RT-PCR were performed using epididymal adipose tissues. HPJ extract up-regulated the levels of phosphorylated adenosine monophosphate-activated protein kinase (AMPK) and its substrate, acetyl-CoA carboxylasse (ACC). HPJ extract also attenuated lipogenic gene expressions of sterol regulatory element-binding protein $1{\alpha}$ (SREBP$1{\alpha}$), fatty acid synthase (FAS), sterol-CoA desaturase 1 (SCD1) and glycerol-3-phosphate acyltransferase (GPAT) in dose-dependent manners. In contrast, expressions of lipolytic genes such as peroxisome proliferator-activated receptor-$\alpha$ (PPAR-${\alpha}$) and CD36, and fatty acid $\beta$-oxidation gene, carnitine palmitoyltransferase-1 (CPT-1) were increased. These results suggest that HPJ extract ameliorates obesity through inhibiting synthesis of lipogenic enzymes as well as stimulating fatty acid oxidation resulting from activation of AMPK, and HPJ extract could be developed as a potential therapeutic agent for obese patients.

Solid Phase Synthesis of Lysine-exposed Peptide-Polymer Hybrids by Atom Transfer Radical Polymerization (ATRP를 이용한 Lysine 말단기를 가진 펩타이드-고분자 하이브리드 합성)

  • Ha, Eun-Ju;Kim, Mijin;Kim, Jinku;An, Seong Soo A.;Paik, Hyun-Jong
    • Polymer(Korea)
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    • v.38 no.4
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    • pp.550-556
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    • 2014
  • Recently, the peptide(or protein)-polymer hybrid materials (PPs) were sought in many research areas as potential building blocks for assembling nanostructures in selective solvents. In PPs, the facile routes of preparing well-defined peptide-polymer bio-conjugates and their specific activities in various applications are important issues. Our strategy to prepare the peptide-polymer hybrid materials was to combine atom transfer radical polymerization (ATRP) method with solid phase peptide synthesis. The standard solid phase peptide synthesis method was employed to prepare the PYGK (proline-tyrosine-glycine-lysine) peptide. PYGK is an analogue peptide, PFGK (proline-phenylalanine-glycine-lysine), which interacted with plasminogen in fibrinolysis. The peptide and the peptide-initiator were characterized with MALDI-TOF mass spectrometry and $^1H$ NMR spectrometer. The peptide-polymer, pSt-PYGK was characterized by GPC, IR, $^1H$ NMR spectrometer and TLC. Spherical micellar aggregates were determined by TEM and SEM. Current synthesis methodology suggested opportunities to create the well-defined peptide-polymer hybrid materials with specific binding activity.

A Study on the Role of Service Design in Creating Resident-driven Safe Community (주민주도형 안전 공동체 조성에 있어 서비스디자인의 역할 탐색)

  • Jeon, Young-Ok
    • Journal of Digital Convergence
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    • v.15 no.6
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    • pp.407-414
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    • 2017
  • With an increase in urban crimes in various forms, this study is intended to analyze the effectiveness of the service design that presented a new model resolving crime risks through differentiated thinking paradigm and problem approaches. The empirical case addressed in this study is 'the project to create resident-driven safe community in Duryu-dong, Dalseo-gu, Deagu though service design'. This project is evaluated as having prepared a prevention-oriented local safety system through a preemptive and resident-centered process. The project was promoted as a 'natural monitoring capacity building program' for residents to prevent local crimes, a 'social role expansion program' for local safety, and 'crime prevention environment design', which provides comprehensive solutions for residents' safety. Here, designers act as exerts in designing a task-based platform that can be driven by residents rather than a visual environment improver, and reorganizing the local ecosystem by expanding the opportunities for residents to interact. This case identifies the role of service design as binding the solidarity of local residents beyond the improvement of the crime environment and giving them the potential capacity to maintain a safe living space in relation to a safety issues of community.

The absorption and excretion times of carp, Cyprinus carpio, treatment with oxolinic acid (향어에서의 oxolinic acid 흡수와 배설속도)

  • Jeong, Hyun-Do;Ha, Jai-Yi;Huh, Min-Do;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.5 no.2
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    • pp.135-142
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    • 1992
  • The absorption and excretion times of oxolinic acid(OX) used in farms as new aquatic antibiotics commonly were evaluated with determination of the effects of water temperature and feeding to parameters by using the bioassay technique. On the same time, antibacterial activity and the complex formation of oxolinic acid with serum proteins of two different fishes were compared to those oxytetracycline(OTC). With more than 10 times lower MIC values than those of OTC in the strains among 13 analyzed fish pathogens. OX did not show the decresed antibacterial activity by the binding of serum proteins in carp and tilapia. It implies more powerful potential of OX as aquatic medicine OTC. The serum concentration of OX after different administrations the oral, i.m., i.v and dipping methods were compared. The higher beginning concentration in serum and faster excretion times were obserbed in i.m. and dipping methods respectively. In the oral and i.m. administration, peak serum concentration after 24-48 hrs and slow excretion times demonstrated in both methods. These pharmacokinetic characteristics similar at $30^{\circ}C$ and $20^{\circ}C$ water temperature conditions, however, beginning serum concentration of OX in fish dipped in $50mg/\ell$ sol after starvation for 2 wks was appeared lower than those of fed fish. It suggests the importance of biological condition of the gill or skin for absorption of antibiotics after dipping administration.

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Optimized Internal Control and Gene Expression Analysis in Epstein-Barr Virus-Transformed Lymphoblastoid Cell Lines

  • Nam, Hye-Young;Kim, Hye-Ryun;Shim, Sung-Mi;Lee, Jae-Eun;Kim, Jun-Woo;Park, Hye-Kyung;Han, Bok-Ghee;Jeon, Jae-Pil
    • Genomics & Informatics
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    • v.9 no.3
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    • pp.127-133
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    • 2011
  • The Epstein-Barr virus-transformed lymphoblastoid cell line (LCL) is one of the major genomic resources for human genetics and immunological studies. Use of LCLs is currently extended to pharmacogenetic studies to investigate variations in human gene expression as well as drug responses between individuals. We evaluated four common internal controls for gene expression analysis of selected hematopoietic transcriptional regulatory genes between B cells and LCLs. In this study, the expression pattern analyses showed that TBP (TATA box-binding protein) is a suitable internal control for normalization, whereas GAPDH (glyceraldehyde-3-phosphate dehydrogenase) is not a good internal control for gene expression analyses of hematopoiesis-related genes between B cells and LCLs at different subculture passages. Using the TBP normalizer, we found significant gene expression changes in selected hematopoietic transcriptional regulatory genes (downregulation of RUNX1, RUNX3, CBFB, TLE1, and NOTCH2 ; upregulation of MSC and PLAGL2) between B cells and LCLs at different passage numbers. These results suggest that these hematopoietic transcriptional regulatory genes are potential cellular targets of EBV infection, contributing to EBV-mediated B-cell transformation and LCL immortalization.

Levels of common salivary protein 1 in healthy subjects and periodontal patients

  • Heo, Seok-Mo;Lee, Sol;Wang, HongTao;Jeong, Jeong Hyeok;Oh, Sang Wook
    • Journal of Periodontal and Implant Science
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    • v.46 no.5
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    • pp.320-328
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    • 2016
  • Purpose: Human saliva, as a vital part of the immune defense system, contains a number of distinct proteins and peptides. Recently human common salivary protein 1 (CSP1) has been identified as an abundant salivary protein and may play a role in promoting the binding of cariogenic bacteria to salivary pellicles. However, nothing else is known regarding the role of CSP1 in periodontology. The aim of this study was to quantify and compare CSP1 levels between healthy subjects and periodontal patients. Methods: This controlled clinical study was conducted in periodontally healthy individuals and patients with chronic periodontitis Chonbuk National University Hospital, with Institutional Review Board approval. Whole saliva samples were collected from 36 healthy subjects and 33 chronic periodontitis patients and analyzed. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immune blotting were conducted to ensure that anti-CSP1 monoclonal antibody (mAb) binds to CSP1 in human saliva. A sandwich enzyme-linked immunosorbent assay (ELISA) system was house-fabricated using mAb-hCSP1#14 and mAb-hCSP1#4 as a capture and a detector mAb, respectively. The CSP1 concentrations in saliva from 36 healthy subjects and 33 periodontal patients were quantified using the CSP1 sandwich ELISA system, and the results were analyzed using the Student's t-test. Results: Immunoblot analysis using mAb-hCSP1 as a probe confirmed that CSP1 in human saliva existed as a single band with a molecular weight of approximately 27-kDa. The quantification of CSP1 concentrations by CSP1 ELISA showed that the median values (25th to 75th percentiles) of periodontal patients and healthy subjects were 9,474 ng/mL (range, 8,434.10,139 ng/mL) and 8,598 ng/mL (range, 7,421.9,877 ng/mL), respectively. The Student's t-test indicated the presence of a statistically significant difference between the 2 groups (P=0.024). Conclusions: The presence of a significant difference in CSP1 levels between healthy subjects and periodontal patients suggests that CSP1 may be a potential biomarker for the detection or screening of periodontitis patients.

BRI3 associates with SCG10 and attenuates NGF-induced neurite outgrowth in PC12 cells

  • Gong, Yanhua;Wu, Jing;Qiang, Hua;Liu, Ben;Chi, Zhikai;Chen, Tao;Yin, Bin;Peng, Xiaozhong;Yuan, Jiangang
    • BMB Reports
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    • v.41 no.4
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    • pp.287-293
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    • 2008
  • In a yeast two-hybrid screen, we identified the microtubule-destabilizing protein SCG10 as a potential effector protein of $BRI_3$. The association was verified using GST pull-down, Co-IP, and their perinuclear co-localization. The analysis of in vitro microtubule polymerization/depolymerization showed that the binding of $BRI_3$ to SCG10 effectively blocked the ability of SCG10 to induce microtubule disassembly, as determined by turbidimetric assays. In intact PC12 cells, $BRI_3$ exhibited the ability to stabilize the microtubule network and attenuate the microtubule-destabilizing activity of SCG10. Furthermore, co-expression of $BRI_3$ with SCG10 attenuated SCG10-mediated PC12 cell neurite outgrowth induced by NGF. These results identify a novel connection between a neuron-specific BRI protein and the cytoskeletal network, suggesting possible roles of BRI3 in the process of neuronal differentiation.

Deubiquitinase Otubain 1 as a Cancer Therapeutic Target (암 치료 표적으로써 OTUB1)

  • Kim, Dong Eun;Woo, Seon Min;Kwon, Taeg Kyu
    • Journal of Life Science
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    • v.30 no.5
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    • pp.483-490
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    • 2020
  • The ubiquitin system uses ligases and deubiquitinases (DUBs) to regulate ubiquitin position on protein substrates and is involved in many biological processes which determine stability, activity, and interaction of the target substrate. DUBs are classified in six groups according to catalytic domain, namely ubiquitin-specific proteases (USPs); ubiquitin C-terminal hydrolases (UCHs); ovarian tumor proteases (OTUs); Machado Joseph Disease proteases (MJDs); motif interacting with Ub (MIU)-containing novel DUB family (MINDY); and Jab1/MPN/MOV34 metalloenzymes (JAMMs). Otubain 1 (OTUB1) is a DUB in the OTU family which possesses both canonical and non-canonical activity and can regulate multiple cellular signaling pathways. In this review, we describe the function of OTUB1 through regulation of its canonical and non-canonical activities in multiple specifically cancer-associated pathways. The canonical activity of OTUB1 inhibits protein ubiquitination by cleaving Lys48 linkages while its non-canonical activity prevents ubiquitin transfer onto target proteins through binding to E2-conjugating enzymes, resulting in the induction of protein deubiquitination. OTUB1 can therefore canonically and non-canonically promote tumor cell proliferation, invasion, and drug resistance through regulating FOXM1, ERα, KRAS, p53, and mTORC1. Moreover, clinical research has demonstrated that OTUB1 overexpresses with high metastasis in many tumor types including breast, ovarian, esophageal squamous, and glioma. Therefore, OTUB1 has been suggested as a diagnosis marker and potential therapeutic target for oncotherapy.

Tyrosinase Inhibitory Activities of Meso-dihydroguaiaretic Acid from Machilus thunbergii (후박나무에서 분리한 Meso-dihydroguaiaretic acid의 tyrosinase 저해활성)

  • Kwon, Hyun Sook;Lee, Kyung Dong;Kim, Su Cheol;Cho, Soo Jeong
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1298-1303
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    • 2015
  • Machilus thunbergii (Lauraceae) is an evergreen tree cultivated in Korea and Japan. M. thunbergii has long been used as a traditional medicine in Korea, China, and Japan to treat various diseases, including edema, abdominal pain, and abdominal distension. In this study, dried stem bark of M. thunbergii extracted in methanol and extract was partitioned into n-hexane, CHCl3, and BuOH. The CHCl3-soluble extracts chromatographed on silica gel column using a CHCl3/acetone and n-hexane/EtOAc mixture to afford Compound 1 and 2. Two dibenzylbutane lignans, macelignan (1) and meso-dihydroguaiaretic acid (2), were isolated from the CHCl3-soluble extract of M. thunbergii stem bark. The structures of 1 and 2 were determined by 1H- and 13C-NMR spectroscopic data analyses and a comparison with literature data. The tyrosinase inhibitory activity of the isolated compounds was evaluated. Among these compounds, Compound 2 strongly inhibited the monophenolase (IC50=10.2 μM) activity of tyrosinase. A kinetic analysis showed that Compound 2 was a competitive inhibitor. The apparent inhibition constant (Ki) for Compound 2 binding to free enzyme was 4.8 μM. Based on these results, it can be concluded that meso-dihydroguaiaretic acid (2) is a potential candidate for the treatment of melanin biosynthesis-related skin diseases.