• Journal of Microbiology and Biotechnology
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• v.31 no.12
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• pp.1716-1721
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• 2021

Chikungunya fever is an arboviral disease caused by the Chikungunya virus (CHIKV). The disease has similar clinical manifestations with other acute febrile illnesses which complicates differential diagnosis in low-resource settings. We aimed to develop a rapid test for CHIKV detection based on the nucleic acid lateral flow immunoassay technology. The system consists of a primer set that recognizes the E1 region of the CHIKV genome and test strips in an enclosed cassette which are used to detect amplicons labeled with FITC/biotin. Amplification of the viral genome was done using open-source PCR, a low-cost open-source thermal cycler. Assay performance was evaluated using a panel of RNA isolated from patients' blood with confirmed CHIKV (n = 8) and dengue virus (n = 20) infection. The open-source PCR-NALFIA platform had a limit of detection of 10 RNA copies/ml. The assay had a sensitivity and specificity of 100% (95% CI: 67.56% - 100%) and 100% (95% CI: 83.89% - 100%), respectively, compared to reference standards of any positive virus culture on C6/36 cell lines and/or qRT-PCR. Further evaluation of its performance using a larger sample size may provide important data to extend its usefulness, especially its utilization in the peripheral healthcare facilities with scarce resources and outbreak situations.

• Biomolecules & Therapeutics
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• v.30 no.4
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• pp.368-379
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• 2022

Hyaluronic acid (HA), a ligand of CD44, accumulates in some types of tumors and is responsible for tumor progression. The nuclear factor erythroid 2-like 2 (NRF2) regulates cytoprotective genes and drug transporters, which promotes therapy resistance in tumors. Previously, we showed that high levels of CD44 are associated with NRF2 activation in cancer stem like-cells. Herein, we demonstrate that HA production was increased in doxorubicin-resistant breast cancer MCF7 cells (MCF7-DR) via the upregulation of HA synthase-2 (HAS2). HA incubation increased NRF2, aldo-keto reductase 1C1 (AKR1C1), and multidrug resistance gene 1 (MDR1) levels. Silencing of HAS2 or CD44 suppressed NRF2 signaling in MCF7-DR, which was accompanied by increased doxorubicin sensitivity. The treatment with a HAS2 inhibitor, 4-methylumbelliferone (4-MU), decreased NRF2, AKR1C1, and MDR1 levels in MCF7-DR. Subsequently, 4-MU treatment inhibited sphere formation and doxorubicin resistance in MCF7-DR. The Cancer Genome Atlas (TCGA) data analysis across 32 types of tumors indicates the amplification of HAS2 gene is a common genetic alteration and is negatively correlated with the overall survival rate. In addition, high HAS2 mRNA levels are associated with increased NRF2 signaling and poor clinical outcome in breast cancer patients. Collectively, these indicate that HAS2 elevation contributes to chemoresistance and sphere formation capacity of drug-resistant MCF7 cells by activating CD44/NRF2 signaling, suggesting a potential benefit of HAS2 inhibition.

• Journal of Sensor Science and Technology
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• v.30 no.4
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• pp.229-235
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• 2021

This study presents a simple approach for the assembly of a free-standing conductive electronic nanofilm of single-walled carbon nanotubes (SWNTs) suitable for enzymatic electrochemical biosensors. A large-scale SWNT electronic film was successfully produced by the dialysis of p-Terphenyl-4,4''-dithiol (TPDT)-treated SWNTs. Furthermore, Horseradish peroxidase (HRP) was immobilized on the TPDT-SWNT electronic film, and the enzymatic detection of hydrogen peroxide (H₂O₂) was demonstrated without mediators. The detection of H₂O₂ in the negative potential range (-0.4 V vs. Ag/AgCl) was achieved by direct electron transfer of heme-based enzymes that were immobilized on the TPDT-SWNT electronic film. The SWNT-based biosensor exhibited a wide detection range of H₂O₂ from 10 µM to 10 mM. The HRP-doped SWNT electronic film achieved a high sensitivity of 342 ㎛A/mM·cm² and excellent selectivity against a variety of redox-active interfering substances, such as ascorbic acid, uric acid, and acetaminophen.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.27 no.4
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• pp.236-245
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• 2024

Purpose: The prevalence of nonalcoholic steatohepatitis (NASH) is increasing with the increasing prevalence of childhood obesity. Although NASH has a high risk of progression to liver fibrosis and cirrhosis, few studies have reported noninvasive markers for predicting hepatic fibrosis in children. This study aimed to evaluate and compare the diagnostic accuracies of serologic biomarkers and scoring systems for hepatic fibrosis in obese children with NASH. Methods: A total of 96 children were diagnosed with NASH based on liver biopsy findings and divided into two groups according to the degree of liver fibrosis: mild (stage 0-1) or advanced (stage 2-4). Clinical and laboratory parameters and serum levels of hyaluronic acid and type IV collagen were measured. The aspartate aminotransferase/platelet ratio index (APRI) and fibrosis-4 (FIB-4) score were calculated. Results: Among the noninvasive markers, only serum type IV collagen level and FIB-4 were significantly different between the two groups. The area under the receiver operating curve of each biomarker and scoring system was 0.80 (95% confidence interval [CI]: 0.70-0.90) for type IV collagen at an optimal cutoff of 148 ng/mL (sensitivity 69.8%, specificity 84.6%), followed by 0.69 (95% CI: 0.57-0.83) for APRI, 0.68 (95% CI: 0.56-0.80) for FIB-4, and 0.65 (95% CI: 0.53-0.77) for hyaluronic acid. Conclusion: Type IV collagen as a single noninvasive serologic biomarker for hepatic fibrosis and FIB-4 as a hepatic fibrosis score are beneficial in predicting advanced hepatic fibrosis and determining proper diagnosis and treatment strategies before fibrosis progresses in obese children with NASH.

• Journal of Radiation Industry
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• v.17 no.2
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• pp.173-181
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• 2023

3D printing is a technology that can transform and process computerized data obtained through modeling or 3D scanning via CAD. In the medical field, studies on customized 3D printing technology for clinical use or patients and diseases continue. The importance of research on filaments and molding methods is increasing, but research on manufacturing methods and available raw materials is not being actively conducted. In this study, we compare the characteristics of each material according to the manufacturing method of the phantom manufactured with 3D printing technology and evaluate its usefulness. We manufactured phantoms of the same size using poly methyl meta acrylate (PMMA), acrylonitrile butadiene styrene (ABS), and Poly Lactic Acid (PLA) based on the international standard phantom of aluminum step wedge. We used SITEC's radiation generator (DigiRAD-FPC R-1000-150) and compared the shielding rate and line attenuation coefficient through the average after shooting 10 times. As a result, in the case of the measured dose transmitted through each phantom, it was confirmed that the appearance of the dose measured for phantoms decreased linearly as the thickness increased under each condition. The sensitivity also decreased as the steps increased for each phantom and confirmed that it was different depending on the thickness and material. Through this study, we confirmed that 3D printing technology can be usefully used for phantom production in the medical field. If further development of printing technology and studies on various materials are conducted, it is believed that they will contribute to the development of the medical research environment.

• Journal of Microbiology and Biotechnology
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• v.34 no.6
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• pp.1222-1228
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• 2024

Protein-specific antibodies are essential for various aspects of protein research, including detection, purification, and characterization. When specific antibodies are unavailable, protein tagging is a useful alternative. Small epitope tags, typically less than 10 amino acids, are widely used in protein research due to the simple modification through PCR and reduced impact on the target protein's function compared to larger tags. The 2B8 epitope tag (RDPLPFFPP), reported by us in a previous study, has high specificity and sensitivity to the corresponding antibody. However, when attached to the C-terminus of the target protein in immunoprecipitation experiments, we observed a decrease in detection signal with reduced immunity and low protein recovery. This phenomenon was not unique to 2B8 and was also observed with the commercially available Myc tag. Our study revealed that C-terminal tagging of small epitope tags requires the addition of more than one extra amino acid to enhance (restore) antibody immunities. Moreover, among the amino acids we tested, serine was the best for the 2B8 tag. Our findings demonstrated that the interaction between a small epitope and a corresponding paratope of an antibody requires an extra amino acid at the C-terminus of the epitope. This result is important for researchers planning studies on target proteins using small epitope tags.

• Journal of fish pathology
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• v.10 no.2
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• pp.125-135
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• 1997

The possibility of identification of families of antibacterial agent residues in fish tissue was studied by disk assay using three test organisms, Bacillus subtilis BGA, Micrococcus luteus ATCC 9341, and Bacillus cereus var. mycoides ATCC 11778. In the present method, a simple clean-up procedure was performed to obtain the aqueous solution from homogenized flounder muscle sample(10g) in Mcilvaine buffer. Then, aqueous solution was fractionated into A and B to be used in disk assay by choloroform and Sep-Pak $C_{18}$ cartridge column after being defatted in hexane. The chloroform layer of fraction A was used for the analysis of macrolide antibiotics(ML), sulfa drugs(SA), chloramphenicol(CP), and quinolone antibiotics(QN). Adsorbed materials to Sep-Pak $C_{18}$ of fraction B were also employed for the analysis of penicillins(PC), tetracyclines(TC), and nitrofuran derivatives(NF) Minimun-detectable concentrations by the present method were, $0.1{\mu}g$/g for oxytetracycline, tetracycline, doxycycline, spiramycin and ciprofloxacin, $0.025{\mu}g$/g for erythromycin and ampicillin, $1.0{\mu}g$/g for sodium nifurstyrenate and florfenical, $0.25{\mu}g$/g for sulfamonomethoxie and sulfadimethoxine, $2.5{\mu}g$/g for oxolinic acid and flumequine, and $15{\mu}g$/g for piromidic acid, respectively. Three test organisms showed different sensitivity patterns for each family of antibacterial agent. Sensitivity patterns were B. cereus > B. subtilis > M. luteus for TC and NF, M. luteus > B, subtilis > B. cereus for ML and PC, B. cereus = B. subtilis > M. luteus for CP and QN, and B. subtilis > B. cereus＝M. luteus for SA. The present method utilizing these characteristics could be useful as a routine screening test for the determination of family of antibacterial agent residues in fish tissue.

• Journal of Chest Surgery
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• v.43 no.4
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• pp.345-355
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• 2010

Background: Extracellular and intracellular pH ($pH_o$ and $pH_i$), which can be changed in various pathological conditions such as hypoxia, affects vascular contractility. To elucidate the mechanism to alter vascular contractility by pH, the effects of pH on reactivity to vasocontracting agents, intracellular $Ca^{2+}$ influx, and $Ca^{2+}$ sensitivity in vascular smooth muscle were examined. Material and Method: Isometric contractions in rat superior mesenteric arteries (SMA) were observed. Intracellular $Ca^{2+}$ concentration ($[Ca^{2+}]_i$) was recorded by microfluorometer using Fura-2/acetoxylmethyl ester in muscle cells. $pH_o$ was increased from 7.4 to 7.8 or decreased to 6.9 or 6.4. $pH_i$ was decreased by applying $NH_4^+$ or propionic acid or modulated by changing $pH_o$ after increasing membrane permeability using $\beta$-escin. Result: Decreases in $pH_o$ from 7.4 to 6.9 or 6.4 shifted concentration-response curve by norepinephrine (NE) or serotonin (SE) to the right and significantly increased half maximal effective concentration (EC50) to NE or SE. Increase in $pH_o$ from 7.4 to 7.8 shifted concentration-response curve by norepinephrine (NE) or serotonin (SE) to the left and significantly reduced EC50 to NE or SE. NE increased $[Ca^{2+}]_i$ in cultured smooth muscle cells from SMA and the increased $[Ca^{2+}]_i$ was reduced by decreases in $pH_o$. NE-induced contraction was inhibited by $NH_4^+$, whereas the resting tension was increased by $NH_4^+$ or propionic acid. When the cell membrane of SMA was permeabilized using ${\beta}$-escin, SMA was contracted by increasing extracellular $Ca^{2+}$ concentration from 0 to $10{\mu}M$ and the magnitude of contraction was decreased by a decrease in $pH_o$ and vice versa. Conclusion: From these results, it can be concluded that a decrease in $pH_o$ might inhibit vascular contraction by reducing the reactivity of vascular smooth muscle to vasoactive agents, $Ca^{2+}$ influx and the sensitivity of vascular smooth muscle to $Ca^{2+}$.

• Childhood Kidney Diseases
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• v.11 no.2
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• pp.220-228
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• 2007

Purpose : We studied the value of clinical signs, laboratory findings and $^{99m}technetium$ dime-rcaptosuccinic acid(DMSA) scan in predicting the presence of vesicoureteral reflux(VUR) in children with first febrile urinary tract infection(UTI). Methods : A retrospective analysis of 84 hospitalized children with first febrile UTI was performed. They underwent DMSA scan and voiding cystourethrography(VCUG) during the acute phase, and were divided into three groups according to the results of the VCUG: absence of VUR, mild(grade I-II) and severe VUR(grade III-V). We studied the relation of VUR to age, gender, fever duration, causative organism, white blood cell count, serum C-reactive protein and result of DMSA scan. Results : Among 84 patients, 6 had mild and 17 had severe VUR. Thirty-eight had abnormal DMSA scan. results Patients with VUR were older than those without VUR(P<0.01). There was a lower probability of infection with Escherichia coli in patients with severe VUR than in those with mild and absent VUR(P<0.01). An abnormal DMSA scan correlated with the presence and severity of VUR(P<0.05). Severe VUR was present in 10.9% of patients with normal DMSA scan. The sensitivity, specificity, positive and negative predictive values of the DMSA scan in predicting the presence of VUR were 69.6%, 63.9%, 42.1%, and 84.8%, respectively. Conclusion : An abnormal DMSA scan correlated to the presence and severity of VUR, but the sensitivity, specificity and positive predictive value of the DMSA scan were low. There-fore, patient with an abnormal DMSA scan requires a VCUG. In order to prevent missing the 10.9% of patients with severe VUR but normal DMSA scans, a VCUG should be performed in patient with normal DMSA scan.

• Analytical Science and Technology
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• v.11 no.5
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• pp.347-352
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• 1998

A study was carried out on the elelctrochemical characteristics of chemically modified electrodes (CMEs) by cyclic voltammetry. Fabrication of CMEs was made by coating with mixed valence (mv) inorganic-metal polymeric films on the glassy carbon electrode surface by potential cycling. Anodic oxidation behavior of methanol and L-ascorbic acid was studied by using CMEs working electrode. Deposition of films such as mv ruthenium oxo/ruthenium cyanide film (mv Ru-O/CN-Ru), mv ruthenium oxo/ferrocyanide film (mv Ru-O/$Fe(CN)_6$), and mv ruthenium oxo/ruthenium cyanide/Rhodium film (mv Ru-O/CN-Ru/Rh) was obtained to coat by scan rate of 50 mV/sec within the specified potential range (-0.5V ~ +1.2V). Film thickness was controlled by the repeat of the potential cycling. Anodic oxidation behavior of methanol was as follow. Calibration graph by using mv Ru-O/CN-Ru film showed linearly from 10 mM to 80 mM MeOH with slope factor of $-7.552{\mu}A/cm^2$. Although slope factor by using mv Ru-O/$Fe(CN)_6$ film was $-5.13{\mu}A/cm^2$, yet linear range of calibration graph could be extended from 10 mM to 100 mM MeOH. Anodic oxidation behavior of L-ascorbic acid was studied by mv Ru-O/CN-Ru film on the glassy carbon electrode and the glassy carbon electrode with Rh film, Glassy carbon electrode modified with Ru polymeric film was showed better sensitivity than the Rh-glassy carbon modified electrode (mv Ru-O/CN-Ru/Rh). Calibration graph was linear from 0.1 mM to 5 mM L-ascorbic acid by using glassy carbon electrode modified with Ru polymeric film. Solpe factor and relative coefficient are $-84.78{\mu}A/mM$ and 0.998, respectively.