• Title/Summary/Keyword: In-vitro diagnosis

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Usefulness of Twinkling Artifacts in Color Doppler Ultrasonography (컬러 도플러 초음파에서 Twinkling artifacts의 유용성)

  • Sim, Hyun-Sun;Kwon, Kyung-Tae
    • The Journal of the Korea Contents Association
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    • v.16 no.10
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    • pp.291-298
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    • 2016
  • The aim of the study was to investigate the diagnostic value of the color Doppler twinkling artifact in disease of urinary system. The intensity of twinkling artifact(TA) with color Doppler was classified into 3 levels, 0(non-TA) to 3(distinct TA). In the in vitro study, acorn jelly with various materials on top was examined using color Doppler at B-mode sonography in a water bath for TA. 31 patients with diagnosis of urinary calculi(renal stones 16, urinary stones 15) based on B-mode sonography were studied in vivo for TA. The materials with rough surfaces such as salt, screw and cubics at B-mode sonography with color Doppler contributed to causing TA. At B-mode sonography without color Doppler 37% of renal stones and 60% of ureter stones were detected. but at B-mode sonography with color Doppler TA was demonstrated for all cases. Superficial roughness of materials affected occurrence of TA at B-mode sonography with color Doppler. Therefore, TA at B-mode sonography without color Doppler could play a role in confident diagnosis of the disease of urinary system.

Antibacterial activity of florfenicol composite nanogels against Staphylococcus aureus small colony variants

  • Liu, Jinhuan;Ju, Mujie;Wu, Yifei;Leng, Nannan;Algharib, Samah Attia;Luo, Wanhe
    • Journal of Veterinary Science
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    • v.23 no.5
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    • pp.78.1-78.13
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    • 2022
  • Background: Florfenicol might be ineffective for treating Staphylococcus aureus small colony variants (SCVs) mastitis. Objectives: In this study, florfenicol-loaded chitosan (CS)-sodium tripolyphosphate (TPP) composite nanogels were prepared to allow targeted delivery to SCV infected sites. Methods: The formulation screening, the characteristics, in vitro release, antibacterial activity, therapeutic efficacy, and biosafety of the florfenicol composite nanogels were studied. Results: The optimized formulation was obtained when the CS and TPP were 10 and 5 mg/mL, respectively. The encapsulation efficiency, loading capacity, size, polydispersity index, and zeta potential of the optimized florfenicol composite nanogels were 87.3% ± 2.7%, 5.8% ± 1.4%, 280.3 ± 1.5 nm, 0.15 ± 0.03, and 36.3 ± 1.4 mv, respectively. Optical and scanning electron microscopy showed that spherical particles with a relatively uniform distribution and drugs might be incorporated in cross-linked polymeric networks. The in vitro release study showed that the florfenicol composite nanogels exhibited a biphasic pattern with the sustained release of 72.2% ± 1.8% at 48 h in pH 5.5 phosphate-buffered saline. The minimal inhibitory concentrations of commercial florfenicol solution and florfenicol composite nanogels against SCVs were 1 and 0.25 ㎍/mL, respectively. The time-killing curves and live-dead bacterial staining showed that the florfenicol composite nanogels were concentration-dependent. Furthermore, the florfenicol composite nanogels displayed good therapeutic efficacy against SCVs mastitis. Biological safety studies showed that the florfenicol composite nanogels might be a biocompatible preparation because of their non-toxic effects on the renal tissue and liver. Conclusions: Florfenicol composite nanogels might improve the treatment of SCV infections.

Preimplantation Genetic Diagnosis in Inborn Error Metabolic Disorders (유전성 대사질환의 착상전 유전진단)

  • Kang, Inn Soo
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.5 no.1
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    • pp.94-107
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    • 2005
  • Prenatal diagnosis (PND) such as amniocentesis or chorionic villi sampling has been widely used in order to prevent the birth of babies with defects especially in families with single gene disorderor chromosomal abnormalities. Preimplantation genetic diagnosis (PGD) has already become an alternative to traditional PND. Indications for PGD have expanded beyond those practices in PND (chromosomal abnormalities, single gene defects), such as late-onset diseases with genetic predisposition, and HLA typing for stem cell transplantation to affected sibling. After in vitro fertilization, the biopsied blastomere from the embryo is analyzed for single gene defect or chromosomal abnormality. The unaffected embryos are selected for transfer to the uterine cavity. Therefore, PGD has an advantage over PND as it can avoid the risk of pregnancy termination. In this review, PGD will be introduced and application of PGD in inborn error metabolic disorder will be discussed.

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Diagnosis of human genetic mutations based on DNA microarray technology

  • Park, Hyun-Gyu
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.17-17
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    • 2005
  • In this presentation, we will discuss several recent achievements developed in my laboratory for microarray-based diagnosis of human genetic mutations including HNF-1 and BRCA1 mutations. To determine the presence of the genetic mutations in a human sample, we prepared allele-specific oligonucleotide chips from selected mutation sites and generated target probes using a tow-step method for Cy-3 DNA $samples^{1)}$ or in vitro transcription of promoter-tagged PCR products for Cy-3 RNA $samples^{2)}$. Hybridization of the target probes to the chips successfully identified all of the genotypes for the tested sites. For more reliable diagnosis, we also employed single base extension (SBE) reaction and zip-code microarray technique for our strategy. Particularly we developed an efficient PNA zip-code microarray for the detection of $HNF-1{\alpha}$ $mutations^{3)}$. Using multiplex SBE reactions and zip-code strategy, we were able to correctly diagnose several mutation sites in exon 2 of $HNF-1{\alpha}$ with a wild-type and mutant including a MODY3 patient. These works represent successful applications of DNA microarray technology for the diagnosis of human genetic mutations.

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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]

In Vitro and Cell Imaging-Based Analysis of Protease Activity Using Nanoparticles (나노입자를 활용한 In vitro 및 세포이미징 기반 단백질분해 효소활성 분석법)

  • Kim, Gae Baik;Kim, Young-Pil
    • Ceramist
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    • v.21 no.3
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    • pp.204-215
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    • 2018
  • Proteases are one of the most abundant classes of enzymes in living organisms and have been considered major targets for drug development. However, despite the ability to specifically cleave their substrates, many attempts to assay protease activity have generally relied upon the use of gel zymography or fluorophore-labeled peptide substrates, which is limited in rapid and multiplex analysis. Here we review the recent advances in nanoparticle (NP)-utilized assays of protease activity focused on in vitro and cell imaging-based approaches. Owing to large surface area and unprecedented physical properties of NPs, these approaches are anticipated to facilitate many applications related to protease activity-based disease diagnosis and drug discovery.

Ginsenoside Rg1 Epigenetically Modulates Smad7 Expression in Liver Fibrosis via MicroRNA-152

  • Rongrong Zhang ;Xinmiao Li ;Yuxiang Gao ;Qiqi Tao;Zhichao Lang;Yating Zhan;Chunxue Li;Jianjian Zheng
    • Journal of Ginseng Research
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    • v.47 no.4
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    • pp.534-542
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    • 2023
  • Background: Ginsenoside Rg1, a bioactive component of Ginseng, has demonstrated anti-inflammatory, anti-cancer, and hepatoprotective effects. It is known that the epithelial-mesenchymal transition (EMT) plays a key role in the activation of hepatic stellate cells (HSCs). Recently, Rg1 has been shown to reverse liver fibrosis by suppressing EMT, although the mechanism of Rg1-mediated anti-fibrosis effects is still largely unclear. Interestingly, Smad7, a negative regulator of the transforming growth factor β (TGF-β) pathway, is often methylated during liver fibrosis. Whether Smad7 methylation plays a vital role in the effects of Rg1 on liver fibrosis remains unclear. Methods: Anti-fibrosis effects were examined after Rg1 processing in vivo and in vitro. Smad7 expression, Smad7 methylation, and microRNA-152 (miR-152) levels were also analyzed. Results: Rg1 significantly reduced the liver fibrosis caused by carbon tetrachloride, and reduced collagen deposition was also observed. Rg1 also contributed to the suppression of collagenation and HSC reproduction in vitro. Rg1 caused EMT inactivation, reducing Desmin and increasing E-cadherin levels. Notably, the effect of Rg1 on HSC activation was mediated by the TGF-β pathway. Rg1 induced Smad7 expression and demethylation. The over-expression of DNA methyltransferase 1 (DNMT1) blocked the Rg1-mediated inhibition of Smad7 methylation, and miR-152 targeted DNMT1. Further experiments suggested that Rg1 repressed Smad7 methylation via miR-152-mediated DNMT1 inhibition. MiR-152 inhibition reversed the Rg1-induced promotion of Smad7 expression and demethylation. In addition, miR-152 silencing led to the inhibition of the Rg1-induced EMT inactivation. Conclusion: Rg1 inhibits HSC activation by epigenetically modulating Smad7 expression and at least by partly inhibiting EMT.

Evaluation of the Anti-Tumor Effects of Paclitaxel-Encapsulated pH-Sensitive Micelles

  • Han, Jong-Kwon;Kim, Min-Sang;Lee, Doo-Sung;Kim, Yoo-Shin;Park, Rang-Woon;Kim, Kwang-Meyung;Kwon, Ick-Chan
    • Macromolecular Research
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    • v.17 no.2
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    • pp.99-103
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    • 2009
  • We evaluated the efficacy of pH-sensitive micelles, formed by methoxy poly(ethylene glycol)-b-poly($\beta$)-amino ester) (PEG-PAE), as carriers for paclitaxel (PIX), a drug currently used to treat various cancers. PTX was successful encapsulated by a film hydration method. Micelles encapsulated more than 70% of the PTX and the size of the PTX-encapsulated micelles (PTX-PM) was less than 150 nm. In vitro experiments indicated that the micelles were unstable below pH 6.5. After encapsulation of PTX within the micelles, dynamic light scattering (DLS) studies indicated that low pH had a similar demicellization effect. An in vitro release study indicated that PTX was slowly released at pH 7.4 (normal body conditions) but rapidly released under weakly acidic conditions (pH 6.0). We demonstrated the safety of micelles from in vitro cytotoxicity tests on HeLa cells and the in vivo anti-tumor activity of PTX-PM in B16F 10 tumor-bearing mice. We concluded that these pH-sensitive micelles have potential as carriers for anti-cancer drugs.

A Paddling Based Locomotive Mechanism for Capsule Endoscopes

  • Park Suk-Ho;Park Hyun-Jun;Park Sung-Jin;Kim Byung-Kyu
    • Journal of Mechanical Science and Technology
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    • v.20 no.7
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    • pp.1012-1018
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    • 2006
  • Diagnosis and treatment using the conventional flexible endoscope in gastro-intestinal tract are very common since advanced and instrumented endoscopes allow diagnosis and treatment by introducing the human body through natural orifices. However, the operation of endoscope is very labor intensive work and gives patients some pains. As an alternative, therefore, the capsule endoscope is developed for the diagnosis of digestive organs. Although the capsule endoscope has conveniences for diagnosis, it is passively moved by the peristaltic waves of gastro-intestinal tract and thus has some limitations for doctor to get the image of the organ and to diagnose more thoroughly. As a solution of these problems, various locomotive mechanisms for capsule endoscopes are introduced. In our proposed mechanism, the capsule-type microrobot has synchronized multiple legs that are actuated by a linear actuator and two mobile cylinders inside of the capsule. For the feasibility test of the proposed microrobot, a series of in-vitro experiments using small intestine without incision were carried out. From the experimental results, our proposed microrobot can advance along the 3D curved and sloped path with the velocity of about $3.29\sim6.26mm/sec$ and $35.1\sim66.7%$ of theoretical velocity. Finally, the proposed locomotive mechanism can be not only applicable to micro capsule endoscopes but also effective to advance inside of gastro-intestinal tract.

Production, Cryopreservation and Transfer of Bovine Embryos Cultured in Serum-Free Medium

  • Hoshi, Hiroyoshi
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.3-7
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    • 2002
  • In vitro embryo culture techniques provide significant contributions not only for a basic research of fertilization and early embryogenesis, but also for a low cost mass production of bovine embryos for transfer, embryo diagnosis, nuclear cloning and the production of transgenic cows. This presentation introduces newly developed serum-free media (IVD101 and IVMD101) that are effective far high yields of transferable embryos of excellent quality from in vitro-matured and fertilized oocytes. Both serum-free media are superior to a conventional serum-containing medium on the increased rates of blastocyst formation, post-thaw embryo viability, and pregnancy after transfer. Furthermore, reduced risks of calf mortality and large calf syndrome are also observed for the serum-free-derived embryos. Serum-derived embryos contain a large number of lipid droplets and immature mitochondria in their cytoplasm that may account for the lower production of transferable embryos and poor embryo quality.

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