• Title/Summary/Keyword: In vivo detection

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Detection of Fish Rhabdoviruses using a Diagnostic Fish Rhabdovirus DNA Chip

  • Kim, Young-Ju;Lee, Myung-Suk
    • Fisheries and Aquatic Sciences
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    • v.8 no.3
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    • pp.185-187
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    • 2005
  • We tested the in vivo ability of a DNA chip to detect virus-specific genes from virus-infected olive flounder Paralichthys olivaceus and rainbow trout Oncorhynchus mykiss. Target cDNA was obtained from total RNA of virus infected cell lines by reverse transcription (RT) and was labeled with fluorescent dye (Cy5-dUTP). The results show the successful detection of infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicaemia virus (VHSV) genes in the virus-infected fishes.

Fluorescent and bioluminescent nanoprobes for in vitro and in vivo detection of matrix metalloproteinase activity

  • Lee, Hawon;Kim, Young-Pil
    • BMB Reports
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    • v.48 no.6
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    • pp.313-318
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    • 2015
  • Matrix metalloproteinases (MMPs) are zinc-dependent endopeptidases that degrade the extracellular matrix (ECM) and regulate the extracellular microenvironment. Despite the significant role that MMP activity plays in cell-cell and cell-ECM interactions, migration, and differentiation, analyses of MMPs in vitro and in vivo have relied upon their abundance using conventional immunoassays, rather than their enzymatic activities. To resolve this issue, diverse nanoprobes have emerged and proven useful as effective activity-based detection tools. Here, we review the recent advances in luminescent nanoprobes and their applications in in vitro diagnosis and in vivo imaging of MMP activity. Nanoprobes with the purpose of sensing MMP activity consist of recognition and detection units, which include MMP-specific substrates and luminescent (fluorescent or bioluminescent) nanoparticles, respectively. With further research into improvement of the optical performance, it is anticipated that luminescent nanoprobes will have great potential for the study of the functional roles of proteases in cancer biology and nanomedicine. [BMB Reports 2015; 48(6): 313-318]

Clinical In Vivo Bio Assay of Glucose in Human Skin by a Tattoo Film Carbon Nano Tube Sensor

  • Ly, Suw Young;Lee, Chang Hyun
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.595-601
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    • 2017
  • In vivo assay of glucose detection was described using a skin tattoo film electrode (STF), and the probe was made from carbon nano tube paste modification film paper. Here in the square-wave stripping anodic working range obtained of $20-100mgL^{-1}$ within an accumulation time of 0 seconds only in sea water electrolyte solutions of pH 7.0. The relative standard deviations of 50 mg glucose that were observed of 0.14 % (n=12), respectively, using optimum stripping accumulation of 30 sec, the low detection limit (S/N) was pegged at 15.8 mg/L. The developed results can be applied to the detect of in vivo skin sensing in real time. Which confirms the results are usable for in vitro or vivo diagnostic clinical analysis.

Functional Imaging of the Multidrug Resistance In Vivo (기능적 영상술을 이용한 다약제 내성의 체내 진단)

  • Lee, Jea-Tae
    • 대한핵의학회:학술대회논문집
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    • 2001.05a
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    • pp.66-75
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    • 2001
  • Although diverse mechanisms are involved in multidrug resistance for chemotherapeutic drugs, the development of cellular P-glycoprotein(Pgp) and multidrug-resistance associated protein (MRP) are important factors in the chemotherapy failure to cancer. Various detection assays provide information about the presence of drug efflux pumps at the mRNA and protein levels. However these methods do not yield information about dynamic function of Pgp and MRP un vivo. Single photon emission tomography (SPECT) and positron emission tomography (PET) are available for the detection of Pgp and MRP-mediated transport. $^{99m}Tc$-sestaMIBl and other $^{99m}Tc$-radiopharmaceuticals are substrates for Pgp and MRP, and have been used in clinical studies for tumor imaging, and to visualize blockade of Pgp-mediated transport after modulation of Pgp pump. Colchicine, verapamil and daunorubicin labeled with $^{11}C$ have been evaluated for the quantification of Pgp-mediated transport with PET in vivo and reported to be feasible substrates with which to image Pgp function in tumors. Leukotrienes are specific substrates for MRP and N-$[^{11}C]$acetyl-leukotriene E4 provides an opportunity to study MRP function non-invasively in vivo. Results obtained from recent publications are reviewed to confirm the feasibility of using SPECT and PET to study the functionality of MDR transporters in vivo.

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Diagnostic Assay of Toxic Zinc in an Ex Vivo Cell Using Voltammetry

  • Ly, Suw-Young;Yoo, Hai-Soo
    • Toxicological Research
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    • v.28 no.2
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    • pp.123-127
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    • 2012
  • Voltammetric detection of the toxic Zn ion was investigated using a fluorine-doped graphite pencil electrode (FPE). It is notable from the study that pencils were used as reference and working electrodes. In all the experiments, a clean seawater electrolyte solution was used to yield good results. The analytical working range was attained to 10 ${\mu}gL^{-1}$. The optimized voltammetric condition was examined to maximize the effect of the detection of trace Zn. The developed sensor was applied to an earthworm's tissue cell. It was found that the methods can be applicable to in vivo fluid or agriculture soil and plant science.

Assay of In Vivo Chromium with a Hollow-fiber Dialysis Sensor

  • Ly, Suw-Young;Yoo, Hai-Soo;Jung, Min-Ki;Ko, Kwang-Hee;Kim, Byung-Jin;Lee, Ki-Chul;Choi, Byung-Min
    • Toxicological Research
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    • v.26 no.3
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    • pp.233-236
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    • 2010
  • The analytical in vivo chromium ion was searched for using a voltammetric hollow-fiber dialysis sensor via square wave stripping voltammetry (SW), cyclic voltammetry (CV), and chronoamperometry. Under optimum parameters, the analytical results indicated linear working ranges of 50~400 mg/l CV and $10{\sim}80\;{\mu}g/l$ SW within a 30-sec accumulation time. The analytical detection limit (S/N) was $6.0\;{\mu}g/l$. The developed method can be applied to in vivo tissues and in ex vivo toxicity assay, as well as to other materials that require chromium analysis.

Fluorescence Detection of Cell Death in Liver of Mice Treated with Thioacetamide

  • Kang, Jin Seok
    • Toxicological Research
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    • v.34 no.1
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    • pp.1-6
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    • 2018
  • The purpose of this study was to detect cell death in the liver of mice treated with thioacetamide (TAA) using fluorescence bioimaging and compare this outcome with that using conventional histopathological examination. At 6 weeks of age, 24 mice were randomly divided into three groups: group 1 (G1), control group; group 2 (G2), fluorescence probe control group; group 3 (G3), TAA-treated group. G3 mice were treated with TAA. Twenty-two hours after TAA treatment, G2 and G3 mice were treated with Annexin-Vivo 750. Fluorescence in vivo bioimaging was performed by fluorescence molecular tomography at two hours after Annexin-Vivo 750 treatment, and fluorescence ex vivo bioimaging of the liver was performed. Liver damage was validated by histopathological examination. In vivo bioimaging showed that the fluorescence intensity was increased in the right upper part of G3 mice compared with that in G2 mice, whereas G1 mice showed no signal. Additionally ex vivo bioimaging showed that the fluorescence intensity was significantly increased in the livers of G3 mice compared with those in G1 or G2 mice (p < 0.05). Histopathological examination of the liver showed no cell death in G1 and G2 mice. However, in G3 mice, there was destruction of hepatocytes and increased cell death. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed many cell death features in the liver of G3 mice, whereas no pathological findings were observed in the liver of G1 and G2 mice. Taken together, fluorescence bioimaging in this study showed the detection of cell death and made it possible to quantify the level of cell death in male mice. The outcome was correlated with conventional biomedical examination. As it was difficult to differentiate histological location by fluorescent bioimaging, it is necessary to develop specific fluorescent dyes for monitoring hepatic disease progression and to exploit new bioimaging techniques without dye-labeling.

In vivo molecular and single cell imaging

  • Hong, Seongje;Rhee, Siyeon;Jung, Kyung Oh
    • BMB Reports
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    • v.55 no.6
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    • pp.267-274
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    • 2022
  • Molecular imaging is used to improve the disease diagnosis, prognosis, monitoring of treatment in living subjects. Numerous molecular targets have been developed for various cellular and molecular processes in genetic, metabolic, proteomic, and cellular biologic level. Molecular imaging modalities such as Optical Imaging, Magnetic Resonance Imaging (MRI), Positron Emission Tomography (PET), Single Photon Emission Computed Tomography (SPECT), and Computed Tomography (CT) can be used to visualize anatomic, genetic, biochemical, and physiologic changes in vivo. For in vivo cell imaging, certain cells such as cancer cells, immune cells, stem cells could be labeled by direct and indirect labeling methods to monitor cell migration, cell activity, and cell effects in cell-based therapy. In case of cancer, it could be used to investigate biological processes such as cancer metastasis and to analyze the drug treatment process. In addition, transplanted stem cells and immune cells in cell-based therapy could be visualized and tracked to confirm the fate, activity, and function of cells. In conventional molecular imaging, cells can be monitored in vivo in bulk non-invasively with optical imaging, MRI, PET, and SPECT imaging. However, single cell imaging in vivo has been a great challenge due to an extremely high sensitive detection of single cell. Recently, there has been great attention for in vivo single cell imaging due to the development of single cell study. In vivo single imaging could analyze the survival or death, movement direction, and characteristics of a single cell in live subjects. In this article, we reviewed basic principle of in vivo molecular imaging and introduced recent studies for in vivo single cell imaging based on the concept of in vivo molecular imaging.

Novel Vectors for the Convenient Cloning and Expression of In Vivo Biotinylated Proteins in Escherichia coli

  • Cho, Eun-Wie;Park, Jung-Hyun;Na, Shin-Young;Kim, Kil-Lyong
    • BMB Reports
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    • v.32 no.5
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    • pp.497-501
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    • 1999
  • Biotinylation of recombinant proteins is a powerful tool for the detection and analysis of proteins of interest in a large variety of assay systems. The recent development of in vivo biotinylation techniques in E. coli has opened new possibilities for the production of site-specifically biotinylated proteins without the need for further manipulation after the isolation of the recombinantly expressed proteins. In the present study, a novel vector set was generated which allows the convenient cloning and expression of proteins of interest fused with an N-terminal in vivo biotinylated thioredoxin (TRX) protein. These vectors were derived from the previously reported pBIOTRX vector into which was incorporated part of the pBluescript II+phagemid multiple cloning site (MCS), amplified by PCR using a pair of sophisticated oligonucleotide primers. The functionality of these novel vectors was examined in this system by recombinant expression of rat transforming growth factor-$\beta$. Western-blot analysis using TRX-specific antibodies or peroxidase-conjugated streptavidin confirmed the successful induction of the fusion protein and the in vivo conjugation of biotin molecules, respectively. The convenience of molecular subcloning provided by the MCS and the effective in vivo biotinylation of proteins of interest makes this novel vector set an interesting alternative for the production of biotinylated proteins.

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Detection of 1270 nm Emission from Singlet Oxygen due to Photodynamic Therapy in vitro and in vivo.

  • Hirano, Toru;Kohno, Eiji;Ito, Toshiaki;Okazaki, Shigetoshi;Hirohata, Toru;Niigaki, Minoru;Kageyama, Kazumi;Miyaki, Sueo
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.515-517
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    • 2002
  • Photodynamic therapy (PDT) is a cancer treatment modality which utilizes the cytotoxicity of the active singlet oxygen derived from irradiation of a tumor accumulated photosensitizer. As the oxygen in the singlet state radiates an emission of 1270nm wavelength when it decays to the triplet state, detection of the emission helps us to understand the mechanism of PDT or to evaluate photosensitizers. We detected the 1270nm emission from photosensitizers Photofrin and ATX-SI0 in vitro and in vivo by means of high sensitive NIR detectors. We obtained the maximum amount of singlet oxygen at irradiation wavelength of 665-670nm from a HeLa tumor in a nude mouse which is injected with ATX-S10.

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