• Journal of Applied Biological Chemistry
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• 제58권3호
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• pp.227-232
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• 2015

A microarray study has been employed to understand changes of gene expression in E. coli KD43162 resistant to ampicillin, ampicillin-sulbactam, piperacillin, piperacillin-tazobactam, cefazolin, cefepime, aztreonam, imipenem, meropenem, gentamicin, tobramycin, ciprofloxacin, levofloxacin, moxifloxacin, fosfomycin, and trimethoprim-sulfamethoxazole except for amikacin using disk diffusion assay. Using Sodium dodecyl sulphate-polyacrylamide gel electrophoresis and MALDI-TOF MS analyses, 36 kDa of outer membrane proteins (OMPs) was found to be deleted in the multidrug resistant E. coli KD 43162. Microarray analysis was used to determine up- and down-regulated genes in relation to multidrug resistant E. coli KD43162. Among the up-regulated genes, these genes were corresponded to express the proteins as penicillin-binding proteins (PBPs), tartronate semialdehyde reductase, ethanolamine utilization protein, shikimate kinase I, allantoinase, predicted SAM-dependent methyltransferase, L-glutamine: D-fructose-6-phosphate aminotransferase (GFAT), phospho-glucosamine mutase, predicted N-acetylmannosamine kinase, and predicted N-acetylmannosamine-6-P epimerase. Up-regulation of PBPs, one of primary target sites of antibiotics, might be responsible for the multidrug resistance in E. coli with increasing amount of target sites. Up-regulation of GFAT enzyme may be related to the up-regulation of PBPs because GFAT produces N-acetylglucosamine, a precursor of peptidoglycans. One of GFAT inhibitors, azaserine, showed a potent inhibition on the growth of E. coli KD43162. In conclusion, up-regulation of PBPs and GFATs with the loss of 36 kDa OMP refers the multidrug resistance in E. coli KD 43162.

• Journal of Microbiology and Biotechnology
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• 제30권11호
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• pp.1640-1650
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• 2020

Colorectal cancer (CRC) is the leading cause of common malignant neoplasm worldwide. Many studies have analyzed compositions of gut microbiota associated with various diseases such as inflammatory bowel diseases (IBD) and colon cancer. One of the most representative bacteria involved in CRC is enterotoxigenic Bacteroides fragilis (ETBF), a species belonging to phylum Bacteroidetes. We used ETBF colonized mice with azoxymethane (AOM)/dextran sulphate sodium (DSS) and zerumbone, a compound with anti-bacterial effect, to determine whether zerumbone could restore intestinal microbiota composition. Four experimental groups of mice were used: sham, ETBF colonized AOM/DSS group, ETBF colonized AOM/DSS group zerumbone 60 mg kg-1 (ETBF/AOM/DSS + Z (60)), and only zerumbone (60 mg kg^-1)-treated group. We performed reversible dye terminators-based analysis of 16S rRNA gene region V3-V4 for group comparison. Microbiota compositions of ETBF/AOM/DSS + Z (60) group and ETBF colonized AOM/DSS group not given zerumbone were significantly different. There were more Bacteroides in ETBF/AOM/DSS + Z (60) group than those in ETBF colonized AOM/DSS group, suggesting that B. fragilis could be a normal flora activated by zerumbone. In addition, based on linear discriminant analysis of effect size (LEfSe) analysis, microbial diversity decreased significantly in the ETBF colonized AOM/DSS group. However, after given zerumbone, the taxonomic relative abundance was increased. These findings suggest that zerumbone not only influenced the microbial diversity and richness, but also could be helpful for enhancing the balance of gut microbial composition. In this work, we demonstrate that zerumbone could restore the composition of intestinal microbiota.

• Bulletin of the Korean Chemical Society
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• 제24권12호
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• pp.1775-1780
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• 2003

A continuous flow system has been developed to determine trace Cu(II), Mn(II), Ni(II) and Zn(II) in a large volume of water samples by a solvent sublation technique. The mixed solution of 1,10-phenanthroline(phen) and thiocyanate ion was used as ligands for the formation of their ternary complexes. The continuous system was constructed in this laboratory with a peristaltic pump, a mini shaker, three mixing bottles and a flotation cell by connecting each part with a polyethylene tube. The flotation conditions such as the flow rate of sample solution and the injection rates of ligand, buffer and surfactant solutions have been investigated to obtain the best sublation efficiencies. Each solution flowed into the flotation cell through each polyethylene tube by the peristaltic pumps. The ternary complexes were floated and extracted into MIBK in a flotation cell of 2 L by bubbling a nitrogen gas. The absorbances of extracted analytes in MIBK were directly measured by graphite furnace-AAS. The concentrations of 1,10-phenanthroline and thiocyanate ion were $2.6\;{\times}\;10^{-3}$ M and $2.3\;{\times}\;10^{-2}$ M in the mixed solution, respectively. The pH of sample solution was adjusted to 5.0 with a buffer solution and 1%(m/v) sodium lauryl sulphate solution was added as a surfactant to support the effective flotation of the complexes. The $N_2$ gas was bubbled at 30 mL/min for 90 minutes for 20 L of sample. Reproducible results of less than 10% RSD and recoveries of 80-120% could be obtained in real samples.

• 한국축산식품학회지
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• 제39권5호
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• pp.844-861
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• 2019

Over the last few years, marine environment was found to be a source of surplus natural products and microorganisms with new bioactive secondary metabolites of interest which can divulge nutritional and biological impact on the host. This study aims to assess the possible, inherent and functional probiotic properties of a novel probiotic strain Enterococcus durans F3 (E. durans F3) isolated from the gut of fresh water fish Catla catla. Parameters for evaluating and describing the probiotics described in FAD/WHO guidelines were followed. E. durans F3 demonstrated affirmative results including simulated bile, acid and gastric juice tolerance with exhibited significant bactericidal effect against pathogens Staphylococcus aureus, Salmonella Typhi, Escherichia coli and Pseudomonas aeruginosa. This can be due to the enterocin produced by E. durans F3 strain, which was resolute by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) gel with amplification of the anticipated fragment of a structural gene; enterocin A, followed by antibiotic susceptibility assessment. Effective antioxidant potentiality against ${\alpha}$-diphenyl-${\alpha}$-picrylhydrazyl free radicals including lipase, bile salt hydrolase activity with auto-aggregation and cell surface hydrophobicity was similarly observed. Results are proving the potentiality of E. durans F3, which can also be used as probiotic starter culture in dairy industries for manufacturing new products that imparts health benefits to the host. Finding the potent and novel probiotic strains will also satisfy the current developing market demand for probiotics.

• 한국축산식품학회지
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• 제41권6호
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• pp.1049-1059
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• 2021

The objective of this study was to evaluate the nutritional content of bullfrog meat from different parts of the animal, including fore-chest, thigh and calf. Bullfrog meat was found to be a rich source of proteins, essential amino acids and minerals, but with a low fat content, compared with other aquatic meat products. There was no significant difference (p>0.05) between thigh and calf in mineral content (K, P, Na, Mg, Ca, Zn, Fe, Cu, and Mn), but the contents of K, P, and Mg were higher in thigh and calf than in the fore-chest (p<0.05). The salt-soluble, water-soluble and insoluble protein bands in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis, from fore-chest, thigh and calf were similar, with the most abundant bands being 35 kDa (salt-soluble protein), 35-48 kDa (water-soluble protein) and 48 kDa (insoluble protein). The results showed that the insoluble protein content in the fore-chest meat was higher than that in the thigh meat and calf meat, but the salt-soluble protein fraction was the most abundant in thigh meat. These results showed that the nutrients in different parts of bullfrog meat were different.

• 농업과학연구
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• 제45권4호
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• pp.615-622
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• 2018

This study was done to determine the optimum sowing time for spring sowing wheat in the southern region of the entire Korean peninsula. Jokyoung and Keumkangmil were sown four times at one-week intervals starting from Feb. 14, 2013. The thousand grain weights of the two wheat varieties were the highest on February 14 with the seed weights ranging from 36.6 to 40 g and significantly decreasing as the seeding time was delayed. The unmatured grain percentage increased sharply when both cultivars were sown starting on March 7. The grain yields were the highest for Keumkangmil with 3.07 MT/ha when sowed on February 21 and 3.37 MT/ha for Jokyoung when sowed on February 14. In both cultivars, the grain yield decreased drastically when they were sown on March 7. Ash content did increase when the sowing date was delayed. The flour gluten index was the highest at 96.2 in Keumkangmil with a February 28 sowing, and the sodium dodecyl sulphate (SDS) segmentation was the lowest ranging from 63.8 to 65.3 mL with the February 28 sowing. The falling number tended to increase with the delay of the sowing period. The flour milling rate was gradually decreased with the delay of sowing, and the bran gradually decreased. When both cultivars were sown after February 28, the grain and flour yields sharply decreased. In the southern region, the optimum time for the spring sowing of wheat is from February 14 to February 21.

• Journal of Electrochemical Science and Technology
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• 제13권2호
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• pp.292-307
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• 2022

The study presents kinetics of degradation and mineralization of an anti-epileptic drug Lamotrigine (LAM) in the aqueous matrix by electrochemical advanced oxidation process (EAOP) on Ti/DSA (Ta₂O₅-Ir₂O₅) and Stainless Steel (SS) anodes using sodium sulphate as supporting electrolyte. On both the anodes, kinetic behaviour was pseudo-first-order for degradation as well as mineralization of LAM. On Ti/DSA anode, maximum LAM degradation of 75.42% was observed at an associated specific charge of 3.1 (Ah/litre) at a current density of 1.38 mA/cm² and 100 ppm Na₂SO₄ concentration. Maximum mineralization attained was 44.83% at an associated specific charge of 3.1 (Ah/litre) at a current density of 1.38 mA/cm² and 50 ppm concentration of Na₂SO₄ with energy consumption of 2942.71 kWh/kgTOC. Under identical conditions on SS anode, a maximum of 98.92% LAM degradation was marked after a specific charge (Q) of 3.1 (Ah/litre) at a current density of 1.38 mA/cm² and 100 ppm concentration of Na₂SO₄. Maximum LAM mineralization on SS anode was 98.53%, marked at a specific charge of 3.1 (Ah/litre) at a current density of 1.38 mA/cm² and 75 ppm concentration of Na₂SO₄, with energy consumption of 1312.17 kWh/kgTOC. Higher Mineralization Current Efficiency (MCE) values were attained for EAOP on SS anode for both degradation and mineralization due to occurrence of combined electro-oxidation and electro-coagulation process in comparison to EAOP on Ti/DSA anode due to occurrence of lone electro-oxidation process.

• Membrane and Water Treatment
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• 제14권2호
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• pp.85-93
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• 2023

Membrane separation is widely used for several applications such as water treatment, membrane reactors and climate change. Cross-linked organic-inorganic hybrid polyvinylidene fluoride (PVDF) / Tetraethyl orthosilicate (TEOS) was adopted for the preparation of optimized hollow membrane (HFM) for membrane distillation or other low pressure separators for mechanical properties and permeability under varying pretreatment schemes. HFMs were prepared on semi-pilot membrane fabrication system. Novel adopted post-treatment schemes involved soaking in glycerol, magnesium sulphate (MgSO₄), sodium hypochlorite (NaOCl), and isopropanol for different durations. All fibers were characterized for morphology using a scanning electron microscope (SEM), surface roughness using atomic force microscope (AFM), elemental composition by examining Energy Dispersive Spectroscopy (EDS), water contact angle (CA°) and porosity. The performance of the fibers was evaluated for pure water permeation flux (PWF). Post-treatment with MgSO₄ gave the highest both tensile modulus and flux. Assessment of properties and performance revealed comparable results with other organic-inorganic separators, HF or flat. In spite of few reported data on post treatment using MgSO₄ in presence of TEOS, this proves the potential of low cost treatment without negative impact on other membrane properties. The flux is also comparable with hypochlorite which manifests substantial precaution requirements in actual industrial use.The relatively high values of flux/bar for sample treated with TEOS, post treated with MgSO₄ and hypochlorite are 88 and 82 LMH/bar respectively.

• Anatomy and Cell Biology
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• 제56권3호
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• pp.382-393
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• 2023

Cell clusters are a histological hallmark feature of intervertebral disc degeneration. Clusters arise from cell proliferation, are associated with replicative senescence, and remain metabolically, but their precise role in various stages of disc degeneration remain obscure. The aim of this study was therefore to investigate small, medium, and large size cell-clusters. For this purpose, human disc samples were collected from 55 subjects, aged 37-72 years, 21 patients had disc herniation, 10 had degenerated non-herniated discs, and 9 had degenerative scoliosis with spinal curvature <45°. 15 non-degenerated control discs were from cadavers. Clusters and matrix changes were investigated with histology, immunohistochemistry, and Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Data obtained were analyzed with spearman rank correlation and ANOVA. Results revealed, small and medium-sized clusters were positive for cell proliferation markers Ki-67 and proliferating cell nuclear antigen (PCNA) in control and slightly degenerated human discs, while large cell clusters were typically more abundant in severely degenerated and herniated discs. Large clusters associated with matrix fissures, proteoglycan loss, matrix metalloproteinase-1 (MMP-1), and Caspase-3. Spatial association findings were reconfirmed with SDS-PAGE that showed presence to these target markers based on its molecular weight. Controls, slightly degenerated discs showed smaller clusters, less proteoglycan loss, MMP-1, and Caspase-3. In conclusion, cell clusters in the early stages of degeneration could be indicative of repair, however sustained loading increases large cell clusters especially around microscopic fissures that accelerates inflammatory catabolism and alters cellular metabolism, thus attempted repair process initiated by cell clusters fails and is aborted at least in part via apoptosis.

• BMB Reports
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• 제38권6호
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• pp.683-689
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• 2005

Antibody to hepatitis B surface antigen (HBsAb) is the important serological marker of the hepatitis B virus (HBV) infection. Conventionally, the hepatitis B surface antigen (HBsAg) obtained from the plasma of HBV carriers is used as the diagnostic antigen for detection of HBsAb. This blood-origin antigen has some disadvantages involved in high cost, over-elaborate preparation, risk of infection, et al. In an attempt to explore the suitable recombinant HBsAg for the diagnostic purpose, the HBV S gene was expressed in Pichia pastoris and the product was applied for detection of HBsAb. Hepatitis B virus S gene was inserted into the yeast vector and the expressed product was analyzed by sodium dodecyl sulphate polyacrolamide gel electrophoresis (SDS-PAGE), immunoblot, electronic microscope and enzyme linked immunosorbent assay (ELISA). The preparations of synthesized S protein were applied to detect HBsAb by sandwich ELISA. The S gene encoding the 226 amino acid of HBsAg carrying ahexa-histidine tag at C terminus was successfully expressed in Pichia pastoris. The His-Tagged S protein in this strain was expressed at a level of about 14.5% of total cell protein. Immunoblot showed the recombinant HBsAg recognized by monoclonal HBsAb and there was no cross reaction between all proteins from the host and normal sera. HBsAb detection indicated that the sensitivity reached 10 mIu (micro international unit)/ml and the specificity was 100% with HBsAb standard of National Center for Clinical Laboratories. A total of 293 random sera were assayed using recombinant S protein and a commercial HBsAb ELISA kit (produced by blood-origin HBsAg), 35 HBsAb positive sera and 258 HBsAb negative sera were examined. The same results were obtained with two different reagents and there was no significant difference in the value of S/CO between the two reagents. The recombinant HBV S protein with good immunoreactivity and specificity was successfully expressed in Pichia pastoris. The reagent for HBsAb detection prepared by Pichia pastoris-derived S protein showed high sensitivity and specificity for detection of HBsAb standard. And a good correlation was obtained between the reagent produced by recombinant S protein and commercial kit produced by blood-origin HBsAg in random samples.