• Title/Summary/Keyword: in vitro detection

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Assessment of Environmental Pollution with Tradescantia Bioassays (자주달개비 생물검정 기법을 이용한 환경오염 평가)

  • Kim Jin Gyu;Sin Hae Sik
    • Proceedings of the Korea Society of Environmental Biology Conference
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    • 2004.05a
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    • pp.1-15
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    • 2004
  • Higher plants can be valuable genetic assay systems for monitoring environmental pollutants and evaluating their biological toxicity. Two assays are considered ideal for in situ monitoring and testing of soil, airborne and aqueous mutagenic agents; the Tradescantia stamen hair assay for somatic cell mutations and the Tradescantia micronucleus assay for chromosome aberrations. Both assays can be used for in vivo and in vitro testing of mutagens. Since higher plant systems are now recognized as excellent indicators and have unique advantages over in situ monitoring and screening, higher plant systems could be accepted by regulatory authorities as an alternative first-tier assay system for the detection of possible genetic damages resulting from the pollutants or chemicals used and produced by industrial sectors. It has been concluded that potential mutagen and carcinogen such as the heavy metals among indoor air particulates, volatile compounds in the working places, soil, and water pollutants contribute to the overall health risk. This contribution can be considerable under certain circumstances. It is therefore important to identify the level of genotoxic activity in the environment and to relate it to the biomarkers of a health risk in humans. The results from the higher plant bioassays could make a significant contribution to assessing the risks of pollutants and protecting the public firom agents that can cause mutation anuor cancer. The plant bioassays, which are relatively inexpensive and easy to handle, are recommended for the scientists who are interested in monitoring pollutants and evaluating their environmental toxicity to living organisms.

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Expression and phosphorylation analysis of soluble proteins and membrane-localised receptor-like kinases from Arabidopsis thaliana in Escherichia coli

  • Oh, Eun-Seok;Eva, Foyjunnaher;Kim, Sang-Yun;Oh, Man-Ho
    • Journal of Plant Biotechnology
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    • v.45 no.4
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    • pp.315-321
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    • 2018
  • Molecular and functional characterization of proteins and their levels is of great interest in understanding the mechanism of diverse cellular processes. In this study, we report on the convenient Escherichia coli-based protein expression system that allows recombinant of soluble proteins expression and cytosolic domain of membrane-localised kinases, followed by the detection of autophosphorylation activity in protein kinases. This approach is applied to regulatory proteins of Arabidopsis thaliana, including 14-3-3, calmodulin, calcium-dependent protein kinase, TERMINAL FLOWER 1(TFL1), FLOWERING LOCUS T (FT), receptor-like cytoplasmic kinase and cytoplasmic domain of leucine-rich repeat-receptor like kinase proteins. Our Western blot analysis which uses phospho-specific antibodies showed that five putative LRR-RLKs and two putative RLCKs have autophosphorylation activity in vitro on threonine and/or tyrosine residue(s), suggesting their potential role in signal transduction pathways. Our findings were also discussed in the broader context of recombinant expression and biochemical analysis of soluble and membrane-localised receptor kinases in microbial systems.

Characteristic Effects of Dangnyo-hwan for Diabetes Control Studied Using LC-MS/MS and ICP (LC-MS/MS 및 ICP를 이용한 당뇨환의 화학적 특성 분석)

  • In, Jeong-do;Im, Dai-sig;Moon, Seung-ho;Ki, Won-ill
    • The Journal of Internal Korean Medicine
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    • v.36 no.3
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    • pp.217-227
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    • 2015
  • Objectives: The primary aim of this study was to identify bioactive compounds in Dangnyo-hwan, a Korean herbal medicine, through instrumental analysis using LC-MS/MS and ICP, and investigate its potential use in diabetes treatment. Methods: The extract of Dangnyo-hwan has 12 medicinal herbs; these were compared with 18 marker substances selected from literature survey. Results: LC-MS/MS analysis could detect 9 of the 18 bioactive compounds: citruline, catalpol, berberine, ginsenoside Rb1, ginsenoside Rg1, oleanolic acid, β-sitosterol, mangiferin, and schizandrin. While harmful heavy metals such as As, Pb, Cd, Hg, Ni, and Cu were not present in high concentrations, Zn concentration was 4.2 mg in 100 g Dangnyo-hwan. Conclusions: Instrumental analysis such as LC-MS/MS and ICP was successfully used to identify bioactive compounds in Dangnyo-hwan. Detection of 9 bioactive substances and Zn from the herb medicine is a valuable finding, and suggests that Dangnyo-hwan is a candidate medicine for diabetes. Further investigations like in vitro assay, percent GPR 119 activity, and percent human DGAT-1 inhibition are underway.

Study on Sex Determination and Detection of Male Specific DNA Band in Bovine IVF Embryos Using Polymerase Chain Reaction (PCR 기법에 의한 소 수정란의 웅성 특이적 DNA Band 출현과 성 판별에 관한 연구)

  • 김현종;오성종;김성우;최화식;윤종택;정구민;임경순
    • Journal of Embryo Transfer
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    • v.11 no.3
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    • pp.283-289
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    • 1996
  • This study was carried out to determine the sex of genomic and embryonic DNA using polymerase chain reaction(PCR). Bovine specific(216bp) and Y chromosome speicific DNA primers(l4lbp) were synthesized and tested for sexing. Bovine embryos used in this study were produced by in vitro fertilization. Few blastomeres for PCR were bisected by nicromanipulator and demi -embryos were cultured in TCM 199 medium containing 0.1% of solcoseryl. The results obtained were as follows; 1. Average optical density of genomic DNA extracted from blood of Hanwoo was 1.79$\pm$ 0.14. 2. 2. The ratio of the demi-embryos developed to blastocyst was 62.1 and 81.9% in morula and blastocyst, respectively. 3. When DNA of 2~4, 5~10 and more than 11 blastomeres was amplified with Y chromosome specific DNA primer by PCR, appreance rate of Y specific DNA band was 16.7, 46.2 and 40.0%, respectively. At least 5 to 10 blastomeres were required to determine the sex of embryos. 4. The rate of demi-embryos developed to blastocyst was 73.3% in TCM 199 medium supplemented with 0.1% solcoceryl. but 55.6% in control.

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Detection of Hydrogen Peroxide in vitro and in vivo Using Peroxalate Chemiluminescent Micelles

  • Lee, Il-Jae;Hwang, On;Yoo, Dong-Hyuck;Khang, Gil-Son;Lee, Dong-Won
    • Bulletin of the Korean Chemical Society
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    • v.32 no.7
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    • pp.2187-2192
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    • 2011
  • Hydrogen peroxide plays a key role as a second messenger in the normal cellular signaling but its overproduction has been implicated in various life-threatening diseases. Peroxalate chemiluminescence is the light emission from a three component reaction between peroxalate, hydrogen peroxide and fluorophores. It has proven great potential as a methodology to detect hydrogen peroxide in physiological environments because of its excellent sensitivity and specificity to hydrogen peroxide. We developed chemiluminescent micelles composed of amphiphilic polymers, peroxalate and fluorescent dyes to detect hydrogen peroxide at physiological concentrations. In this work, we studied the relationship between the chemiluminescence reactivity and stability of peroxalate by varying the substitutes on the aryl rings of peroxalate. Alkyl substitutes on the aryl ring of peroxalate increased the stability against water hydrolysis, but diminished the reactivity to hydrogen peroxide. Chemiluminescent micelles encapsulating diphenyl peroxalate showed significantly higher chemiluminescence intensity than the counterpart encapsulating dimethylphenyl or dipropylphenyl peroxalate. Diphenyl peroxalate-encapsulated micelles could detect hydrogen peroxide generated from macrophage cells stimulated by lipopolysaccharide (LPS) and image hydrogen peroxide generated during LPS-induced inflammatory responses in a mouse.

Effects of Static Magnetic Fields of Earphones and Headphones on Pacemakers and Implantable Cardioverter Defibrillators (이어폰 및 헤드폰의 정자기장이 인공심장 박동기 및 이식형 제세동기에 미치는 영향)

  • Chung, J.W.;Choi, S.B.;Park, J.S.;Kim, D.W.
    • Journal of Biomedical Engineering Research
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    • v.36 no.1
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    • pp.31-36
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    • 2015
  • In this study, we evaluated the effects of static magnetic fields of earphones and headphones on pacemakers and implantable cardioverter defibrillators(ICDs). Five pacemakers and three ICDs were subjected to in-vitro test with three headphones which were an in-ear earphone, clip-on headphone, and closed-back headphone. Each implantable device was placed in close proximity(within 3 mm) to the ear-pad of each of the earphone and headphones for 3 min. As a result, no effects were observed on the pacemakers for the earphone and headphones during the test, but an effect was observed on one ICD for the clip-on and closed-back headphone during the test. When the ICD was placed in close proximity to the headphones, the ICD temporarily suspended functions of tachyarrhythmia detection and therapy. The effect was not observed in this study when the headphones were at least 2 cm from the ICD. Based on these findings, patients with ICDs should be advised to keep earphones and headphones at least 2 cm apart from their ICDs.

Large Scale Directed Assembly of SWNTs and Nanoparticles for Electronics and Biotechnology

  • Busnaina, Ahmed;Smith, W.L.
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2011.10a
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    • pp.9-9
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    • 2011
  • The transfer of nano-science accomplishments into technology is severely hindered by a lack of understanding of barriers to nanoscale manufacturing. The NSF Center for High-rate Nanomanufacturing (CHN) is developing tools and processes to conduct fast massive directed assembly of nanoscale elements by controlling the forces required to assemble, detach, and transfer nanoelements at high rates and over large areas. The center has developed templates with nanofeatures to direct the assembly of carbon nanotubes and nanoparticles (down to 10 nm) into nanoscale trenches in a short time (in seconds) and over a large area (measured in inches). The center has demonstrated that nanotemplates can be used to pattern conducting polymers and that the patterned polymer can be transferred onto a second polymer substrate. Recently, a fast and highly scalable process for fabricating interconnects from CMOS and other types of interconnects has been developed using metallic nanoparticles. The particles are precisely assembled into the vias from the suspension and then fused in a room temperature process creating nanoscale interconnect. The center has many applications where the technology has been demonstrated. For example, the nonvolatile memory switches using (SWNTs) or molecules assembled on a wafer level. A new biosensor chip (0.02 $mm^2$) capable of detecting multiple biomarkers simultaneously and can be in vitro and in vivo with a detection limit that's 200 times lower than current technology. The center has developed the fundamental science and engineering platform necessary to manufacture a wide array of applications ranging from electronics, energy, and materials to biotechnology.

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A Monoclonal Antibody That Specifically Binds Chitosan In Vitro and In Situ on Fungal Cell Walls

  • Schubert, Max;Agdour, Siham;Fischer, Rainer;Olbrich, Yvonne;Schinkel, Helga;Schillberg, Stefan
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1179-1184
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    • 2010
  • We report the generation of the first monoclonal antibody that specifically binds to the polysaccharide chitosan. Mice were immunized with a mixture of chitosans, and hybridoma clones were screened for specific binders, resulting in the isolation of a single clone secreting a chitosan-specific IgM, mAbG7. In ELISAs, the antibody could bind to chitosans of varying composition, but demonstrated the highest affinity for chitosans with lower degrees of acetylation (DA) and very poor binding to chitin. We tested the ability of the antibody to bind to chitosan in situ, using preparations of fungal cell walls. Immunofluorescence microscopy confirmed that the antibody bound strongly to the cell walls of fungi with high levels of chitosan, whereas poor staining was observed in those species with cell walls of predominantly chitin or cellulose. The potential use of this antibody for the detection of fungal contamination and the protection of plants against fungal pathogens is discussed.

Uterine infarction in a patient with uterine adenomyosis following biochemical pregnancy

  • Lee, Jae-Yeon;Hwang, Kyu-Ri;Won, Kyu-Hee;Lee, Da-Yong;Jeon, Hye-Won;Moon, Min-Hwan
    • Clinical and Experimental Reproductive Medicine
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    • v.41 no.4
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    • pp.174-177
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    • 2014
  • Adenomyosis is a common gynecological disorder characterized by the presence of endometrial glands and stroma deep within the myometrium associated with myometrial hypertrophy and hyperplasia. Focal uterine infarction after IVF-ET in a patient with adenomyosis following biochemical pregnancy has not been previously reported, although it occurs after uterine artery embolization in order to control symptoms caused by fibroids or adenomyosis. We report a case of a nulliparous woman who had uterine adenomyosis presenting with fever, pelvic pain and biochemical abortion after undergoing an IVF-ET procedure and the detection of a slightly elevated serum hCG. Focal uterine infarction was suspected after a pelvic magnetic resonance imaging demonstrated preserved myometrium between the endometrial cavity and inner margin of the necrotic myometrium. This case demonstrates that focal uterine infarction should be considered in the differential diagnosis of acute abdominal pain, vaginal bleeding and infectious signs in women experiencing biochemical abortion after an IVF-ET procedure.

Production and Evaluation of Monoclonal Antibodies Against Recombinant Coat Protein of Lily mottle virus for Western Blotting and Immono-blot Analysis

  • Chung, Bong-Nam;Yoon, Ju-Yeon;Choi, Gug-Sun
    • The Plant Pathology Journal
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    • v.25 no.3
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    • pp.225-230
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    • 2009
  • Lily mottle virus (LMoV) causes flower quality reduction in Lilium spp. The coat protein gene was RT-PCR-amplified from total RNA extracted from infected lily leaves and the amplified fragment was cloned into the pRSET expression vector tagged with a His-MBP. The plasmid of recombinant coat protein was used to transform an Escherichia coli strain pLysS and was expressed. The coat protein was purified by affinity chromatography using a Ni-NTA resin. The identity of the purified protein was confirmed by SDS-PAGE. The in vitro-expressed protein was used for immunization of mice. The polyclonal and monoclonal antibodies reacted specifically for the detection of LMoV in lily extracts in Western blot. Moreover the monoclonal antibodies reacted with lily extracts in DAS-ELISA with no unspecific or heterologous reactions against other non-serologically related viruses, but the polyclonal antibodies revealed a weak reaction against both infected lily and healthy control.