• Title/Summary/Keyword: Biomedical Laboratory

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Development of an Escherichia coli Biofilm Model on Transwell®

  • Kim, Bok Yung;Thyiam, General;Kang, Ji-Eun;Lee, Seung-Hwan;Park, Sang-Hee;Kim, Jung-Sun;Abraham, Marion
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.3
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    • pp.112-117
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    • 2012
  • Escherichia coli biofilm, reported to be produced in the human intestine causing a significant health risk, was successfully grown on transwell$^{(R)}$. This biofilm layer was identified by crystal violet staining and prepared for the in vitro E. coli biofilm system which can be used to screen for inhibitors. The biofilm formation did not show a change in transepithelial electrical resistance values. Furthermore, rhodamine 123 staining showed that the dye did not pass through the membrane once biofilm was formed.

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Microplate hybridization assay for detection of isoniazid resistance in Mycobacterium tuberculosis

  • Han, Hye-Eun;Lee, In-Soo;Hwang, Joo-Hwan;Bang, Hye-Eun;Kim, Yeun;Cho, Sang-Nae;Kim, Tae-Ue;Lee, Hye-Young
    • BMB Reports
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    • v.42 no.2
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    • pp.81-85
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    • 2009
  • Early and accurate detection of drug resistant Mycobacterium tuberculosis can improve both the treatment outcome and public health control of tuberculosis. A number of molecular-based techniques have been developed including ones using probe molecules that target drug resistance-related mutations. Although these techniques are highly specific and sensitive, mixed signals can be obtained when the drug resistant isolates are mixed with drug susceptible isolates. In order to overcome this problem, we developed a new drug susceptibility test (DST) for one of the most effective anti-tuberculosis drug, isoniazid. This technique employed a microplate hybridization assay that quantified signals from each probe molecule, and was evaluated using clinical isolates. The evaluation analysis clearly showed that the microplate hybridization assay was an accurate and rapid method that overcame the limitations of DST based on conventional molecular techniques.

In-Situ Observation of New Extra-Vascular Threadlike Structure of Mouse Using a Fluorescence Stereoscopic Microscope

  • Sung, Baeck-Kyoung;Lee, Ja-Woong;Lee, Byung-Cheon;Johng, Hyun-Min;Baik, Ku-Youn;Nam, Tae-Jeong;Park, Dae-Hun;Soh, Kyeong-Sun;Soh, Kwang-Sup
    • Journal of Pharmacopuncture
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    • v.7 no.3
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    • pp.73-76
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    • 2004
  • We report the new threadlike structures outside the blood vessels of mice. For this, we developed an in-situ searching method of the structure by vital staining with the dye of acridine orange and using a fluorescent stereomicroscope designed specifically for this purpose. We consider that the newly found threadlike structure might be rediscovery of the extra-vascular Bonghan duct which was reported in 1963 by Bonghan Kim.

Inhibitory Effects of Epigallocatechin-3-Gallate on Microsomal Cyclooxygenase-1 Activity in Platelets

  • Lee, Dong-Ha;Kim, Yun-Jung;Kim, Hyun-Hong;Cho, Hyun-Jeong;Ryu, Jin-Hyeob;Rhee, Man Hee;Park, Hwa-Jin
    • Biomolecules & Therapeutics
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    • v.21 no.1
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    • pp.54-59
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    • 2013
  • In this study, we investigated the effect of (-)-epigallocatechin-3-gallate (EGCG), a major component of green tea catechins from green tea leaves, on activities of cyclooxygenase (COX)-1 and thromboxane synthase (TXAS), thromboxane $A_2$ ($TXA_2$) production associated microsomal enzymes. EGCG inhibited COX-1 activity to 96.9%, and TXAS activity to 20% in platelet microsomal fraction having cytochrome c reductase (an endoplasmic reticulum marker enzyme) activity and expressing COX-1 (70 kDa) and TXAS (58 kDa) proteins. The inhibitory ratio of COX-1 to TXAS by EGCG was 4.8. These results mean that EGCG has a stronger selectivity in COX-1 inhibition than TXAS inhibition. In special, a nonsteroid anti-inflammatory drug aspirin, a COX-1 inhibitor, inhibited COX-1 activity by 11.3% at the same concentration ($50{\mu}M$) as EGCG that inhibited COX-1 activity to 96.9% as compared with that of control. This suggests that EGCG has a stronger effect than that of aspirin on inhibition of COX-1 activity. Accordingly, we demonstrate that EGCG might be used as a crucial tool for a strong negative regulator of COX-1/$TXA_2$ signaling pathway to inhibit thrombotic disease-associated platelet aggregation.

Real-time Nucleic Acid Sequence Based Amplification (Real-time NASBA) for Detection of Norovirus

  • Lee, In-Soo;Choi, Dong-Hyuk;Lim, Jae-Won;Cho, Yoon-Jung;Jeong, Hye-Sook;Cheon, Doo-Sung;Bang, Hye-Eun;Jin, Hyun-Woo;Choi, Yeon-Im;Park, Sang-Jung;Kim, Sung-hyun;Lee, Hye-Young;Kim, Tae-Ue
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.191-196
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    • 2011
  • Noroviruses (noroV) are the major cause of nonbacterial gastroenteritis in humans worldwide. Since noroV cannot yet be cultured in vitro and their diagnosis by electron microscopy requires at least $10^6$ viral particles/g of stool a variety of molecular detection techniques represent an important step towards the detection of noroV. In the present study, we have applied real-time nucleic acid sequence-based amplification (real-time NASBA) for simultaneous detection of NoroV genogroup I (GI) and genogroup II (GII) using standard viral RNA. For real-time NASBA assay which can detected noroV GI and GII, a selective region of the genes encoding the capsid protein was used to design primers and genotype-specific molecular beacon probes. The specificity of the real-time NASBA using newly designed primers and probes were confirmed using standard viral RNA of noroV GI and GII. To determine the sensitivity of this assay, serial 10-fold dilutions of standard viral RNA of noroV GI and GII were used for reverse transcription polymerase chain reaction (RT-PCR) and real-time NASBA. The results showed that while agarose gel electrophoresis could detect RT-PCR products with 10 pg of standard viral RNA, the real-time NASBA assay could detect 100 fg of standard viral RNA. These results suggested that the real-time NASBA assay has much higher sensitivity than conventional RT-PCR assay. This assay was expected that might detect the viral RNA in the specimens which could have been false negative by RT-PCR. There were needed to perform real-time NASBA with clinical specimens for evaluating accurate sensitivity and specificity of this assay.

Efficacy of glycine powder air-polishing in supportive periodontal therapy: a systematic review and meta-analysis

  • Zhu, Mengyuan;Zhao, Meilin;Hu, Bo;Wang, Yunji;Li, Yao;Song, Jinlin
    • Journal of Periodontal and Implant Science
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    • v.51 no.3
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    • pp.147-162
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    • 2021
  • Purpose: This systematic review and meta-analysis was conducted to assess the effects of glycine powder air-polishing (GPAP) in patients during supportive periodontal therapy (SPT) compared to hand instrumentation and ultrasonic scaling. Methods: The authors searched for randomized clinical trials in 8 electronic databases for relevant studies through November 15, 2019. The eligibility criteria were as follows: population, patients with chronic periodontitis undergoing SPT; intervention and comparison, patients treated by GPAP with a standard/nozzle type jet or mechanical instrumentation; and outcomes, bleeding on probing (BOP), patient discomfort/pain (assessed by a visual analogue scale [VAS]), probing depth (PD), gingival recession (Rec), plaque index (PI), clinical attachment level (CAL), gingival epithelium score, and subgingival bacteria count. After extracting the data and assessing the risk of bias, the authors performed the meta-analysis. Results: In total, 17 studies were included in this study. The difference of means for BOP in patients who received GPAP was lower (difference of means: -8.02%; 95% confidence interval [CI], -12.10% to -3.95%; P<0.00001; I2=10%) than that in patients treated with hand instrumentation. The results of patient discomfort/pain measured by a VAS (difference of means: -1.48, 95% CI, -1.90 to -1.06; P<0.001; I2=83%) indicated that treatment with GPAP might be less painful than ultrasonic scaling. The results of PD, Rec, PI, and CAL showed that GPAP had no advantage over hand instrumentation or ultrasonic scaling. Conclusions: The findings of this study suggest that GPAP may alleviate gingival inflammation more effectively and be less painful than traditional methods, which makes it a promising alternative for dental clinical use. With regards to PD, Rec, PI, and CAL, there was insufficient evidence to support a difference among GPAP, hand instrumentation, and ultrasonic scaling. Higher-quality studies are still needed to assess the effects of GPAP.

Soybean Peptides Induce Apoptosis in HeLa Cells by Increasing Oxidative Stress

  • Sung, Ho Joong;Jeong, Yu Jin;Kim, Jihyun;Jung, Eunsun;Jun, Jin Hyun
    • Biomedical Science Letters
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    • v.21 no.2
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    • pp.77-83
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    • 2015
  • Soy proteins have been extensively studied because of its multiple health benefits. However, the effects of soy proteins on human cervical cancer cells are still unclear. Therefore, this study investigated the effects of soy proteins on HeLa cells and human fibroblasts by using soybean peptides (SPs). SPs selectively increased the generation of reactive oxygen species and apoptosis in HeLa cells but not in fibroblasts. In addition, SPs suppressed the migration of HeLa cells. Although the molecular mechanisms underlying the effects of SPs on human cervical cancer cells need to be investigated further, our findings provide insights on the therapeutic effects of soy protein on cervical cancer.

Parkin-induced Decrease of ${\beta}$-catenin is Mediated by Protein Kinase C in TNF-${\alpha}$-treated HeLa Cells

  • Lee, Min Ho;Jung, Byung Chul;Kim, Sung Hoon;Lee, Juyeon;Jung, Dongju;Cho, Jang-Eun;Rhee, Ki-Jong;Kim, Yoon Suk
    • Biomedical Science Letters
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    • v.19 no.2
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    • pp.83-89
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    • 2013
  • Parkin is a protein known to have tumor suppressive functions. In a previous study, we determined that Parkin expression restores susceptibility to TNF-${\alpha}$-induced death in HeLa cells. ${\beta}$-catenin is a key protein in the Wnt signaling pathway and excessive activation of the ${\beta}$-catenin pathway can promote cancer development. In this study, we found that ${\beta}$-catenin levels decreased dramatically in Parkin over-expressing HeLa cells treated with TNF-${\alpha}$. We used chemical inhibitors of cell signaling pathways to identify the signaling molecules involved in ${\beta}$-catenin down-regulation. Our results indicate that the PKC inhibitor (RO-31-7549) blocked parkin-induced down-regulation of ${\beta}$-catenin. We also show that Parkin-induced decrease in cell viability in TNF-${\alpha}$-treated HeLa cells is alleviated upon treatment with a PKC inhibitor. Taken together, these results suggest the possibility that ${\beta}$-catenin reduction may be associated with Parkin-induced decrease of cell viability in TNF-${\alpha}$ treated HeLa cells.

The Progression of SARS Coronavirus 2 (SARS-CoV2): Mutation in the Receptor Binding Domain of Spike Gene

  • Sinae Kim;Jong Ho Lee;Siyoung Lee;Saerok Shim;Tam T. Nguyen;Jihyeong Hwang;Heijun Kim;Yeo-Ok Choi;Jaewoo Hong;Suyoung Bae;Hyunjhung Jhun;Hokee Yum;Youngmin Lee;Edward D. Chan;Liping Yu;Tania Azam;Yong-Dae Kim;Su Cheong Yeom;Kwang Ha Yoo;Lin-Woo Kang;Kyeong-Cheol Shin;Soohyun Kim
    • IMMUNE NETWORK
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    • v.20 no.5
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    • pp.41.1-41.11
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    • 2020
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is a positive-sense single-stranded RNA (+ssRNA) that causes coronavirus disease 2019 (COVID-19). The viral genome encodes twelve genes for viral replication and infection. The third open reading frame is the spike (S) gene that encodes for the spike glycoprotein interacting with specific cell surface receptor - angiotensin converting enzyme 2 (ACE2) - on the host cell membrane. Most recent studies identified a single point mutation in S gene. A single point mutation in S gene leading to an amino acid substitution at codon 614 from an aspartic acid 614 into glycine (D614G) resulted in greater infectivity compared to the wild type SARS-CoV2. We were interested in investigating the mutation region of S gene of SARS-CoV2 from Korean COVID-19 patients. New mutation sites were found in the critical receptor binding domain (RBD) of S gene, which is adjacent to the aforementioned D614G mutation residue. This specific sequence data demonstrated the active progression of SARS-CoV2 by mutations in the RBD of S gene. The sequence information of new mutations is critical to the development of recombinant SARS-CoV2 spike antigens, which may be required to improve and advance the strategy against a wide range of possible SARS-CoV2 mutations.

Cardamonin Inhibits the Expression of Inducible Nitric Oxide Synthase Induced by TLR2, 4, and 6 Agonists

  • Kim, Ah-Yeon;Shim, Hyun-Jin;Kim, Su-Yeon;Heo, Sung-Hye;Youn, Hyung-Sun
    • Biomedical Science Letters
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    • v.24 no.2
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    • pp.102-107
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    • 2018
  • Toll-like receptors (TLRs) play an important role for host defense against invading pathogens. The activation of TLRs signaling leads to the activation of $NF-{\kappa}B$ and the expression of pro-inflammatory gene products such as cytokines and inducible nitric oxide synthase (iNOS). To evaluate the therapeutic potential of cardamonin, which is a naturally occurring chalcone from Alpinia species (zingiberaceous plant species), $NF-{\kappa}B$ activation and iNOS expression induced by MALP-2 (TLR2 and TLR6 agonist) or LPS (TLR4 agonist) were examined. Cardamonin inhibited the activation of $NF-{\kappa}B$ induced by MALP-2 or LPS. Cardamonin also suppressed the iNOS expression induced by MALP-2 or LPS. These results suggest that cardamonin has the specific mechanism for anti-inflammatory responses by regulating of TLRs signaling pathway.