• Journal of Microbiology and Biotechnology
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• v.31 no.11
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• pp.1481-1489
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• 2021

Early and accurate detection of pathogens is important to improve clinical outcomes of bloodstream infections (BSI), especially in the case of drug-resistant pathogens. In this study, we aimed to develop a culture-independent digital PCR (dPCR) system for multiplex detection of major sepsis-causing gram-negative pathogens and antimicrobial resistance genes using plasma DNA from BSI patients. Our duplex dPCR system successfully detected nine targets (five bacteria-specific targets and four antimicrobial resistance genes) through five reactions within 3 hours. The minimum detection limit was 50 ag of bacterial DNA, suggesting that 1 CFU/ml of bacteria in the blood can be detected. To validate the clinical applicability, cell-free DNA samples from febrile patients were tested with our system and confirmed high consistency with conventional blood culture. This system can support early identification of some drug-resistant gram-negative pathogens, which can help improving treatment outcomes of BSI.

• Journal of Life Science
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• v.25 no.12
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• pp.1458-1469
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• 2015

Many organic solvent-tolerant bacteria have been isolated from all environments such as soil, waste-water, even deep sea after first isolation report of organic solvent-tolerant bacterium. Most organic solvent- tolerant isolates have been determined to be Gram-negative bacteria, because Gram-negative bacteria have inherent tolerance property toward hostile organic solvents more than Gram-positive bacteria. The mechanisms of organic solvent tolerance have been elucidated extensively using mainly organic solvent-tolerant Gram-negative bacteria. The solvent-tolerance mechanisms in Gram-positive bacteria can be found in comparatively recent research. Organic solvents exhibited different toxicity depending on the solvent, and the tolerance levels of organic solvent-tolerant bacteria toward organic solvents were also highly changeable among species and strains. Therefore, organic solvent-tolerant bacteria could coped with solvent toxicity and adapted to solvent stress through the multifactorial and multigenic adaptative strategies. They could be survived even in the hyper concentrations of organic solvents by mechanisms which include: changes in cell morphology and cell behaviour, cell surface modifications, cell membrane adaptations, solvent excretion pumps, chaperones and anti-oxidative response. The aim of this work is to review the representative solvent tolerant bacteria and the adaptative and tolerance strategies toward organic solvents in organic solvent-tolerant bacteria, and their potential industrial and environmental impact.

• Journal of Microbiology and Biotechnology
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• v.28 no.5
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• pp.671-678
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• 2018

Papiliocin, isolated from the swallowtail butterfly (Papilio xuthus), is an antimicrobial peptide with high selectivity against gram-negative bacteria. We previously showed that the N-terminal helix of papiliocin (PapN) plays a key role in the antibacterial and anti-inflammatory activity of papiliocin. In this study, we measured the selectivity of PapN against multidrug-resistant gram-negative bacteria, as well as its anti-inflammatory activity. Interactions between Trp2 of PapN and lipopolysaccharide (LPS), which is a major component of the outer membrane of gram-negative bacteria, were studied using the Trp fluorescence blue shift and quenching in LPS micelles. Furthermore, using circular dichroism, we investigated the interactions between PapN and LPS, showing that LPS plays critical roles in peptide folding. Our results demonstrated that Trp2 in PapN was buried deep in the negatively charged LPS, and Trp2 induced the ${\alpha}$-helical structure of PapN. Importantly, docking studies determined that predominant electrostatic interactions of positively charged arginine residues in PapN with phosphate head groups of LPS were key factors for binding. Similarly, hydrophobic interactions by aromatic residues of PapN with fatty acid chains in LPS were also significant for binding. These results may facilitate the development of peptide antibiotics with anti-inflammatory activity.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.54 no.4
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• pp.377-387
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• 2021

Fecal contamination levels of discharge water from inland pollution sources were investigated in Iwon-myeon (Taean-gun), South Korea. Gram-negative bacteria were isolated during the investigation and the antimicrobial resistance patterns of the isolates were examined to estimate their impact on the coastal environment. The ranges of total coliform and fecal coliform of 12 samples from four major inland pollution sources were 79-490,000 MPN/100 mL and 2.0-490,000 MPN/100 mL, respectively, with the highest level of fecal contamination at Station No. 3. A total of 137 strains (14 genus) were isolated, of which 86 strains (62.8%) were Enterobacteriaceae. The identified isolates were as follows: Pseudomonas spp. (35 strains), Klebsiella spp. (20 strains), Serratia spp. (20 strains), and Escherichia spp. (19 strains). The isolated Gram-negative bacteria showed the highest antimicrobial resistance to ampicillin (81.8%), followed by amoxicillin/clavulanic acid (64.2%), ceftiofur (61.3%), and cefoxitin (59.1%). Antimicrobials in which less than 10% of isolates showed antimicrobial resistance were ciprofloxacin (3.6%) and gentamicin (2.2%). Resistance to one or more antimicrobials was observed in 121 strains (88.3%) and 84 strains (61.3%) showed a tendency for multiple antimicrobial resistance.

• KSBB Journal
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• v.26 no.6
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• pp.543-551
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• 2011

Bacterial antibiotic resistance is a real and growing problem for both Gram positive and Gram negative bacterial pathogens in the hospital setting. Among Gram negative bacteria, the ubiquitous bacterium Pseudomonas aeruginosa is a particular concern in immunocompromised and burn patients. The present study evaluated antibacterial activity and efficacy of a Korean herbal medicine against eight multi-drug resistant clinical isolates of P. aeruginosa (0225, 0254, 0347, 0826, 1113, 1378, 1731, and 2492) isolated at Daegu Catholic University Hospital. Methanol extracts of Galla rhois (5 and 10 mg/mL) displayed inhibition diameters for isolate 2492 of 10 and 12 mm, respectively, in a conventional disc diffusion assay. In seven kinds of Korean herbal medicines, increased inhibitory power of Lonicera japonica, Gardenia jasminoides, Galla rhois, and Scultellaria baicalensis was evident with the fermentation of six kinds of lactic acid bacteria. Three lactic acid bacteria (Lactobacillus plantarum subsp. plantarum KCTC 3108, L. casei KCTC 3109, and L. fermentum KCTC 3112) were identified as excellent strains for the production of antibacterial materials. In the six Korean herbal medicine extracts, strong inhibitory activity of fermented Forsythia suspensa, Glycyrrhizae radix, Lycium chinense, Platycodon grum, and Schizandra chinensis with five kinds of lactic acid bacteria was evident for seven multi-drug resistant P. aeruginosa isolates.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.307-308
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• 2005

Bioconverted hydroxy fatty acid from ${\gamma}-linolenic$ acid showed antibacterial activity against Gram-positive bacteria such as Bacillus subtilis (ATCC 6633), Listeria monocytogenes (ATCC 19166), Staphylococcus aureus (ATCC 6538) and S. aureus (KCTC 1916) and one Gram-negative bacteria, Pseudomonas aeruginosa (KCTC 2004) with MIC ranging from 250 to $750\;{\mu}g/ml$ against five of eleven bacteria tested.

• Journal of the Korean Magnetic Resonance Society
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• v.19 no.2
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• pp.54-60
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• 2015

Papiliocin, from the swallowtail butterfly, Papilio xuthus, shows high bacterial cell selectivity against Gram-negative bacteria. Recently, we designed a 22mer analog with N-terminal helix from $Lys^3$ to $Ala^{22}$, PapN. It shows outstanding antimicrobial activity against Gram-negative bacteria with low toxicity against mammalian cells. In this study, we determined the 3-D structure of PapN in 300 mM DPC micelle using NMR spectroscopy and investigated the interactions between PapN and DPC micelles. The results showed that PapN has an amphipathic ${\alpha}$-helical structure from $Lys^3$ to $Lys^{21}$. STD-NMR and DOSY experiment showed that this helix is important in binding to the bacterial cell membrane. Furthermore, we tested antibacterial activities of PapN in the presence of salt for therapeutic application. PapN was calcium- and magnesium-resistant in a physiological condition, especially against Gram-negative bacteria, implying that it can be a potent candidate as peptide antibiotics.

• Advances in Traditional Medicine
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• v.7 no.3
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• pp.286-289
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• 2007

Heracleum nepalense D Don. (Umbelliferae) is a small shrub having high glabrescent stem found in stream banks in Sikkim. Various medicinal properties which include antidiarrhoeal, antiseptic, anti-influenzal etc. have been attributed for this plant in the traditional system of medicine in Sikkim. In present investigation the methanol extract of roots of Heracleum nepalense was subjected for its effectiveness against both Gram-positive and Gram-negative bacteria causing diarrhoea. The roots extract was tested for its minimum inhibitory concentration (MIC) against both Gram-positive and Gram-negative organisms causing diarrhoea. Further, the zones of inhibition produced by the crude extract against few sensitive strains was measured and compared with those of standard antibiotic ciprofloxacin. It is evident that the methanol extract is very active against the bacteria causing diarrhoea at low concentrations. The antibacterial efficacy of the root extract was found to decrease in the following order against different tested bacterial strains like Shigella dysenteriae, Escherichia coli, Shigella boydii, Vibrio cholerae, Salmonella typhimurium.

• Natural Product Sciences
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• v.13 no.4
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• pp.279-282
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• 2007

The aim of the present study was to investigate the various chemical components present in the cone volatile oil of Cupressus macrocarpa and also determine its antimicrobial activity. Totally 13 compounds were identified with 99.99% by GC-MS analysis. The major compounds identified were terpinene-4-ol (19.42%), dinopol (15.63%), ${\alpha}$-pinene (13.58%), and ${\beta}$-pinene (12.16%). The antimicrobial activity was carried out for the oil and a 2% cream formulation using cup plate method by measuring the zone of inhibition. The gram positive organisms used were Bacillus subtilis, Staphylococcus aureus, Bacillus megaterium, and Bacillus cogulans. The gram negative organisms used were Escherichia coli, Kleibseilla pneumonia, Pseudomonas aeruginosa and Salmonella typhi. In vitro antifungal studies were also carried out by using organisms, Candida albicans, Aspergillus flavus, Trichoderma lignorum and Cryptococcus neoformans. The standard drugs used were penicillin ($100{\mu}g/mL$), gentamycin ($100{\mu}g/mL$) and griseofulvin ($100{\mu}g/mL$) for gram positive bacteria, gram negative bacteria and fungi respectively. Both oil and cream formulation showed good activity against fungi than bacteria. This study is being reported for the first time on cone volatile oil of this plant.

• Journal of Aquaculture
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• v.14 no.4
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• pp.221-226
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• 2001

The ethanol extract of root bark of Morus alba strongly inhibited the Gram positive bacteria like Streptococcus sp., Lactococcus garvieae and Staphylococcus sp., but weakly the Gram negative bacteria like Listonella anguillarm and Edwardsiella tarda. It was more effective in liquid medium than in solid medium. The minimum inhibition concentration (MIC) of the extract in liquid medium was 19.8 and 790~1185 $\mu\textrm{g}$/$m\ell$ for the Gram positive and Gram negative bacteria, respectively. The extract concentration, at which the growth was totally inhibited, was 67.2~403.0 $\mu\textrm{g}$/$m\ell$ for the Gram positive bacteria but it was as high as 1185 $\mu\textrm{g}$/$m\ell$ for L. anguillarum and almost ineffective against E. tarda. For diet supplementation of the extract, effective soaking duration was 3 minutes. The fish diet soaked in the extract inhibited the growth of all the tested Gram positive strains, but not the Gram negative strains. The relationship between the weight of fish diet and absorption of the extract by the fish diet was Y=7.5757X + 4.6962($R^2$ = 0.9998).