• Title/Summary/Keyword: Molecular biological monitoring

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Real-time FRET imaging of cytosolic FAK signal on microwavy patterned-extracellular matrix (ECM) (미세파상 패턴 ECM 에서 세포질 FAK 신호의 실시간 FRET 이미징)

  • Suh, Jung-Soo;Jang, Yoon-Kwan;Kim, Tae-Jin
    • Journal of Biomedical Engineering Research
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    • v.40 no.1
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    • pp.1-6
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    • 2019
  • Human mesenchymal stem cells (hMSC) are multipotent stromal cells that have great potential to differentiate into a variety of cell types such as osteocytes, chondrocytes, and myocytes. Although there have been many studies on their clinical availability, little is known about how intracellular signals can be modulated by topographic features of the extracellular matrix (ECM). In this study, we investigated whether and how microwavy-patterned extracellular matrix (ECM) could affect the signaling activity of focal adhesion kinase (FAK), a key cellular adhesion protein. The fluorescence resonance energy transfer (FRET)-based FAK biosensor-transfected cells are incubated on microwavy-patterned surfaces and then platelet derived growth factor (PDGF) are treated to trigger FAK signals, followed by monitoring through live-cell FRET imaging in real time. As a result, we report that PDGF-induced FAK was highly activated in cells cultured on microwavy-patterned surface with L or M type, while inhibited by H type-patterned surface. In further studies, PDGF-induced FAK signals are regulated by functional support of actin filaments, microtubules, myosin-related proteins, suggesting that PDGF-induced FAK signals in hMSC upon microwavy surfaces are dependent on cytoskeleton (CSK)-actomyosin networks. Thus, our findings not only provide new insight on molecular mechanisms on how FAK signals can be regulated by distinct topographical cues of the ECM, but also may offer advantages in potential applications for regenerative medicine and tissue engineering.

Investigation of fish community structure and species diversity in two river estuary ecosystems, the Taehwa River and Changwon Stream, based on conventional survey and eDNA metabarcoding (어구조사 및 환경 DNA 메타바코딩을 이용한 태화강, 창원천 하구 생태계의 어류 군집 구조 및 종 다양성 평가)

  • Hee-kyu Choi;Yu Rim Kim;Soon Young Hwang;Yeounsu Chu;Pyoungbeom Kim;Hyuk Je Lee
    • Korean Journal of Environmental Biology
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    • v.41 no.4
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    • pp.637-656
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    • 2023
  • River estuaries are dynamic and productive ecosystems with high regional biodiversity. Environmental DNA (eDNA) has become a useful approach to assessing biodiversity in aquatic ecosystems. This study was conducted to investigate fish community characteristics and species diversity in two river estuary ecosystems, the Taehwa River and Changwon Stream. We further compared conventional and eDNA metabarcoding analyses of the fish communities. The conventional survey was performed in May, July, and October 2022, while the eDNA analysis was conducted only in May. We observed various fish species with different life histories, including carp, goby, and marine fish. We also found that migratory fish, such as dace Tribolodon hakonensis, sweetfish Plecoglossus altivelis, and eel Auguilla japonica, occurred in the Taehwa River, suggesting high river connectivity. Marine fish species were predominant in the Changwon Stream, as this river is located close to the sea. The diversity indices showed that the Taehwa River generally had higher species richness, evenness, and diversity values than the Changwon Stream. A total of 9-19 species were detected in the conventional survey for the three sites, whereas 11-18 species were found from eDNA analysis. The findings indicate that the sensitivity of eDNA was similar to or higher than that of the conventional method. Our study findings suggest the efficiency and efficacy of eDNA-based fish community monitoring, although with some shortcomings in applying the genetic marker to Korean fish, including no clear-cut distinction for Korean endemic species and/or genetically closely related species groups.

Molecular Imaging of Arthritis in the Angiogenic Vasculature Using A 123I-Vascular Endothelial Growth Factor Receptor Antibody

  • Kim, Sung-Min;Choi, Na-Eun;Song, Young-Kyu;Cho, Gyung-Goo;Bang, Jeong-Kyu;Kim, Sang-Mi;Lee, Sang-Hoon;Ryu, Eun-Kyoung
    • Bulletin of the Korean Chemical Society
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    • v.33 no.6
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    • pp.1890-1894
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    • 2012
  • Vascular endothelial growth factor (VEGF) and its receptor (VEGFR) have been implicated in the pathogenesis of rheumatoid arthritis, which is angiogenesis dependent. Antibody-based molecular imaging improves targeting, and antibody radiolabeling is useful for monitoring biological events $in$ $vivo$ $via$ PET or SPECT. We investigated the potential of molecular imaging to diagnose arthritis with VEGFR-2 $in$ $vivo$. The $^{123}I$-VEGFR-2 antibody was prepared by the iodogen tube method. The radioligand was injected into arthritic mice, and micro SPECT/CT was performed. The arthritic mice were examined by 4.7-T MRI and immunohistochemistry. The $^{123}I$-VEGFR-2 antibody showed high uptake in the arthritic region at 1 h postinjection on SPECT/CT but no uptake in the control animals after radioligand injection. In MR images, the arthritic tissue of the mice was correlated with regions labeled by the $^{123}I$-VEGFR-2 antibody. Immunohistochemical localization showed markedly increased expression of VEGFR-2 in the endothelial cells, fibroblasts, and macrophages of the arthritic mice.

Ttrosine Hydroxylase in Japanese Medaka (Oryzias latipes): cDNA Cloning and Molecular Monitoring of TH Gene Expression As a Biomarker (송사리 Tyrosine Hydroxylase: cDNA 클로닝 및 생물지표로서의 TH 유전자 발현의 분자생물학적 추적)

  • Shin, Sung-Woo;Kim, Jung-Sang;Chon, Tae-Soo;Lee, Sung-Kyu;Koh, Sung-Cheol
    • Environmental Analysis Health and Toxicology
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    • v.15 no.4
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    • pp.131-137
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    • 2000
  • The release of hazardous waste materials into the environment poses serious risks in humans and ecosystems. The risk assessment of environmental pollutants including hazardous chemicals requires a comprehensive measurement of hazard and exposure of the chemicals that can be achieved by toxicity evaluation using a biological system such as biomarkers. In this report we have tried to develop a biomarker used to elucidate a molecular basis of, and to monitor abnormal behaviors caused by diazinon in Japanese medaka (Oryzias latipes) as a model organism. First, an attempt was made to clone tyrosine hydroxylase gene from Japanese medaka that would be a candidate for a biomarker for neuronal modulations and behaviors. For monitoring experiments at behavioral and molecular biological levels, the fish were treated under different sublethal conditions of diazinon and their behavioral responses were observed . In this study we have successfully cloned a partial TH gene from the medaka fish through PCR screening of an ovary cDNA library. DNA sequencing analysis revealed that the amplified fragment was 327 bp encoding 109 amino acids. Comparing the DNA sequence of medaka TH with other species, TH gene revealed the DNA sequence was completely identical to that of rat TH. In the RT-PCR, 330 Up of mRNA was consistently amplified in all the treated samples including control There were no significant differences in the TH expression level regardless of treating concentrations (1∼5,000 ppb) and time (0∼48 hr) The reason appeared to be that RT-PCR was not performed using through a quantitative analysis normalized against an actin gene expression. Organ or tissue - specific detection of TH activity and mRNA as biomarkers will be a useful monitoring tool for neurobehavioral changes in fish influenced by toxic chemicals. Furthermore, quantitative analysis of locomotive patterns and its correlation with the neurochemical and molecular data would be highly useful in measuring toxicity and hazard ofvarious environmental pollutants.

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Molecular Monitoring of Eukaryotic Plankton Diversity at Mulgeum and Eulsukdo in the Lower Reaches of the Nakdong River (낙동강 하류 물금과 을숙도 수환경의 진핵 플랑크톤 종조성에 대한 분자모니터링)

  • Lee, Jee Eun;Lee, Sang-Rae;Youn, Seok-Hyun;Chung, Sang Ok;Lee, Jin Ae;Chung, Ik Kyo
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.17 no.3
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    • pp.160-180
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    • 2012
  • We have studied the eukaryotic plankton species diversity to compare the community structure of fresh and brackish waters in the lower reaches of the Nakdong River using metagenomic methods. We constructed 18S rDNA clone libraries of total DNAs extracted from environmental water samples collected at Mulgeum (MG100929, fresh) and Eulsukdo bridge (ES, brackish). Through the steps of colony PCR, PCR-RFLP, sequencing and similarity analysis, we discovered the diverse species composition of eukaryotic plankton. Total 338 clones (170 at MG100929 and 168 at ES) were analyzed, and then we found 74 phylotypes (49 for MG100929 and 25 for ES). From the phylogenetic analysis, we confirmed various eukaryotic plankton of broad range of taxonomic groups, including Stramenopiles, Cryptophyta, Viridiplantae, Alveolata, Rhizaria, Metazoa, and Fungi. We also found several unreported species in Korea and candidates of new taxonomic entities at levels higher than genus. Especially, the cryptic species diversity including unreported phylotypes of Pirsonia (Stramenopiles) and Perkinsea (Alveolata) suggests that the molecular monitoring method can produce new informative biological data in monitoring the changes in the Nakdong River Mouth ecosystem.

Forisome based biomimetic smart materials

  • Shen, Amy Q.;Hamlington, B.D.;Knoblauch, Michael;Peters, Winfried S.;Pickard, William F.
    • Smart Structures and Systems
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    • v.2 no.3
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    • pp.225-235
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    • 2006
  • With the discovery in plants of the proteinaceous forisome crystalloid (Knoblauch, et al. 2003), a novel, non-living, ATP-independent biological material became available to the designer of smart materials for advanced actuating and sensing. The in vitro studies of Knoblauch, et al. show that forisomes (2-4 micron wide and 10-40 micron long) can be repeatedly stimulated to contract and expand anisotropically by shifting either the ambient pH or the ambient calcium ion concentration. Because of their unique abilities to develop and reverse strains greater than 20% in time periods less than one second, forisomes have the potential to outperform current smart materials as advanced, biomimetic, multi-functional, smart sensors or actuators. Probing forisome material properties is an immediate need to lay the foundation for synthesizing forisomebased smart materials for health monitoring of structural integrity in civil infrastructure and for aerospace hardware. Microfluidics is a growing, vibrant technology with increasingly diverse applications. Here, we use microfluidics to study the surface interaction between forisome and substrate and the conformational dynamics of forisomes within a confined geometry to lay the foundation for forisome-based smart materials synthesis in controlled and repeatable environment.

Development of EST-SSR markers for the Korean endemic species Chrysosplenium aureobracteatum (Saxifragaceae)

  • SHIN, Jae-Seo;KIM, Bo-Yun;KIM, Yong-In;LEE, Jung-Hoon;KIM, Young-Dong
    • Korean Journal of Plant Taxonomy
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    • v.50 no.1
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    • pp.22-26
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    • 2020
  • Chrysosplenium aureobracteatum Y. I. Kim & Y. D. Kim (Saxifragaceae) is a recently described endemic species growing in the central part of the Korean peninsula. It requires constant monitoring for conservation due to its limited distributions. There is also a need for molecular markers for proper assessments of the genetic differentiation of C. aureobracteatum from species morphologically similar to it. In this study, we developed microsatellite markers that can be used to evaluate the genetic diversity of this species, representing fundamental data with which to conserve the natural populations of the species. A total of 17 expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed by the Illumina pair-end sequencing of the transcriptomes of C. aureobracteatum. These markers were successfully applied to populations of C. aureobracteatum and to its most closely related species, C. barbatum, revealing high polymorphism in both species. The EST-SSR markers developed in this study were proven to be useful not only to monitor the population genetic structure of C. aureobracteatum for conservation purposes but also to study the genetic delimitation of the species from species closely related to it.

Phytomonitoring of the Genotoxicity of Environmental Pollutants: An Application to Armenian Nuclear Power Plant

  • Kim, Jin Kyu;Aroutiounian, Rouben M.;Nebish, Anna A.;Kim, Jin-Hong
    • Journal of Radiation Industry
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    • v.9 no.4
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    • pp.181-185
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    • 2015
  • Today the biosafety evaluation, a common problem of vital importance, is based on internationally proved test-systems, standards and techniques. The paradigm of biosafety includes multidisciplinary approach, a combination of physical, chemical and biological tests to monitor the environmental level of pollutants and needs to be improved by modern approaches. The genetic risk of environmental pollutions has long been studied by many researchers. In this study, used was the known sensitive plant test-system, clones of plant Tradescantia (spiderwort) able to detect gene mutations (frequency of mutational events and formation of micronuclei) in combination with chemical and, in some instances, with radiological measurements. In addition, male gametophyte generation of fruit trees was applied as bioindicators of genotoxicity. The obtained results did not show any significant increase along with wind direction. As for the male gametophyte assay, the fertility of the investigated fruit-trees near to NPP did not significantly differ from that of the control point. The influence of the NPP on the male generative system of the investigated taxa of fruit trees for the investigated year was not revealed. The system described needs to be expanded by species of interest (human) as there is a difficulty to transfer the revealed dose correlations to humans. The development of this idea includes various levels: population (epidemiological studies), individual, cellular, molecular (DNA), etc.

Review of Nanoparticles in Drinking Water: Risk Assessment and Treatment (나노입자의 현황조사 및 처리방안 마련을 위한 문헌연구)

  • Kim, Seung-Hyun;Hong, Seung-kwan;Yoon, Je-Yong;Kim, Doo-Il;Lee, Sang-Ho;Kweon, Ji-Hyang;Kim, Hyung-Soo;ko, Seok-Dock;Kuk, Ji-Hoon
    • Journal of Korean Society of Water and Wastewater
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    • v.25 no.2
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    • pp.201-212
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    • 2011
  • Nanotechnology is the applied science which develops new materials and systems sized within 1 to 100 nanometer, and improves their physical, chemical, and biological characteristics by manipulating on an atomic and molecular scale. This nanotechnology has been applied to wide spectrum of industries resulting in production of various nanoparticles. It is expected that more nanoparticles will be generated and enter to natural water bodies, imposing great threat to potable water resources. However their toxicity and treatment options have not been throughly investigated, despite the significant growth of nanotechnology-based industries. The objective of this study is to provide fundamental information for the management of nanoparticles in water supply systems through extensive literature survey. More specifically, two types of nanoparticles are selected to be a potential problem for drinking water treatment. They are carbon nanoparticles such as carbon nanotube and fullerene, and metal nanoparticles including silver, gold, silica and titanium oxide. In this study, basic characteristics and toxicity of these nanoparticles were first investigated systematically. Their monitoring techniques and treatment efficiencies in conventional water treatment plants were also studied to examine our capability to mitigate the risk associated with nanoparticles. This study suggests that the technologies monitoring nanopartilces need to be greatly improved in water supply systems, and more advanced water treatment processes should be adopted for better control of these nanoparticles.

Development of a Reporter System Monitoring Regulated Intramembrane Proteolysis of the Transmembrane bZIP Transcription Factor ATF6α

  • Kim, Jin-Ik;Kaufman, Randal J.;Back, Sung Hoon;Moon, Ja-Young
    • Molecules and Cells
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    • v.42 no.11
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    • pp.783-793
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    • 2019
  • When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of $ATF6{\alpha}$ by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human $ATF6{\alpha}$ N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-$hATF6{\alpha}$ deletion variant was cleaved to liberate active transcription activator encompassing GV-$hATF6{\alpha}$ fragment which could translocate into the nucleus. The translocated GV-$hATF6{\alpha}$ fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-$hATF6{\alpha}$(333) represents an innovative tool to investigate regulated intramembrane proteolysis of $ATF6{\alpha}$. It can substitute active pATF6(N) binding motif-based reporter cell lines.