• Journal of Applied Biological Chemistry
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• v.58 no.3
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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.

• Analytical Science and Technology
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• v.8 no.4
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• pp.561-568
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• 1995

Two methods, precipitation and ultrafiltration, were applied in order to recover platinum group metals(PGM) by complexing them with water-soluble polyelectrolytes, e.g., polyethyleneimine [PEl], poly(2-vinylpyridine) [2-PVP], poly (4-vinylpyridine) [4-PVP], and poly (styrene sulfonic acid) [PSSA]. In the precipitation method, the PGM-polyelectrolyte complex that was formed by mixing first with polybase, e.g.,4-PVP at pH 1 was precipitated by further mixing with polyacid, e.g., PSSA. However, the recovery of PGM obtained by this method was not quantitative(less than 70%). The "sandwiching" binding between the metal anions and two polyelectrolytes was examined by X-ray photoelectron spectroscopy(XPS). The XPS studies indicated that the PGM atom was bound with the acdic and basic polyelectrolyte via its oxygen and nitrogen atom, respectively. The recovery of PGM using polyelectrolyte was further studied by ultrafiltration methods as follows : The PGM ions, eomplexed at pH 1 with polyelectrolyte, allowed the applicntion of membrane filtration by virtue of the great differences in molecular weights between PGM and other low molecular weight species. By applying this method, Pd and Pt (ca. $10^{-4}M$) were selectively separated almost quantitatively from coexisting metal ions, e.g., $Cu^{2+}$ and $Ni^{2+}$. The EPR spectra and viscosity measurements indicated that these polyelectrlytes were not bound to $Cu^{2+}$ and $Ni^{2+}$ ions at this pH, which provided the basis for selective separation of PGM(Pd, Pt and Rh) from these coexisting ions.

• Applied Chemistry for Engineering
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• v.24 no.1
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• pp.49-54
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• 2013

We fabricated a composite carbon electrode to remove nitrate ions selectively from a mixed solution of anions. The electrode was fabricated by coating the surface of a carbon electrode with the nitrate-selective anion exchange resin (BHP55, Bonlite Co.) powder. We performed capacitive deionization (CDI) experiments on a mixed solution containing chloride, nitrate, and sulfate ions using a BHP55 cell constructed with the fabricated electrode. The removal of nitrate ions in the BHP55 cell was compared to that of a membrane capacitive deionization (MCDI) cell constructed with ion exchange membranes. The total quantity of ions adsorbed in BHP55 cell was $38.3meq/m^2$, which is 31% greater than that of MCDI cell. In addition, the number of nitrate adsorption in the BHP55 cell was $15.9meq/m^2$ (42% of total adsorption), 2.1 times greater than the adsorption in the MCDI cell. The results showed that the fabricated composite carbon electrode is very effective in the selective removal of nitrate ions from a mixed solution of anions.

• Analytical Science and Technology
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• v.8 no.3
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• pp.221-227
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• 1995

The fiber optic fluorosensor that shows a specific selectivity for calcium ion is studied. This sensor employs protein Calmodulin(CaM) which forms a fluorescent chelate with $Ca^{2+}$. A dialysis membrane is used to entrap a fluorescein isothiocyanate-labeled CaM solution at the common end of a bifurcated fiber optic bundle. The sensing mechanism of this sensor is based on the shifts in the fluorescence spectrum of metal-calmodulin complexes which FCaM forms a chelate with $Ca^{2+}$. Upon binding with $Ca^{2+}$, CaM undergoes a conformational change which induces a change in the fluorescence of FCaM. This change in fluorescence signal which is measured by photomultiflier tube is related to the concentration of $Ca^{2+}$ for calibration curve. Detection limit for $Ca^{2+}$ and the interference effects by $Mg^{2+}$, $Eu^{3+}$ and $La^{3+}$ for this sensor are studied. Response time and life time for this fluorosensor are also investigated.

• Mass Spectrometry Letters
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• v.11 no.2
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• pp.36-40
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• 2020

Untargeted metabolomics is a useful tool for drug development focusing on novel chemotherapeutic and chemopreventative agents against cancer cells. In recent years, quadrupole time of flight liquid chromatography-mass spectrometry (Q-TOF LC/MS)-based untargeted metabolomic approaches have gained importance to evaluate the effect of these agents at the molecular level. The researchers working on cell culture studies still do not apply standardized methodologies on sample preparation for untargeted metabolomics approaches. In this study, the rough and wet lysis techniques performed on MCF-7 breast cancer cells were compared with each other via the Q-TOF LC/MS-based metabolomic approach. The C18 and hydrophilic interaction liquid chromatography (HILIC) columns were used for the separation of the metabolites in MCF-7 cell lysates. 505 peaks were detected through the HILIC column and 551 peaks were found through the C18 column for the wet lysis technique. This situation supported by the base peak chromatograms showed that the wet lysis technique allowed us to extract higher number of non-polar metabolites. Almost equal number of metabolites was found for the C18 and HILIC columns (697 peaks for the HILIC column and 695 peaks for the C18 column) when the rough lysis technique was used. However, the intensities of polar metabolites were higher for the rough lysis technique on base peak chromatograms for both the HILIC and C18 columns. Although cell lysis technique, which is the first step in the sample preparation for cell culture studies, does not cause dramatic differences in the number of the detected metabolite peaks, it affects the polar and non-polar metabolite ratio significantly. Therefore, it must be considered carefully especially for in vitro drug development studies.

• Applied Biological Chemistry
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• v.26 no.1
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• pp.58-64
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• 1983

The fatty acid composition of phospholipid from the leaves of rice plants grown at $28^{\circ}C$ and harvested at the 3-4 leaf stage was determined for 8 cultivars. Change among cultivars observed in the composition has been found to be correlative to the chilling susceptivity of the plants at the given leaf-stage. The chilling-resistant cultivars contain phospholipid with higher degree of unsaturation and larger relative proportion of unsaturated fatty acids than the chilling-sensitive ones. The indices for unsaturation of phospholipid from a cultic-ar are well in accordance with its resistance to cold damage. clearly demonstrating that the fluidity of biomembrane which is generally regarded as the prerequisite for a cell to maintain its membrane-related physiological activity at a low temperature is exclusively controlled by the fatty acid composition of phospholipid. Also identified were the components of phospholipid, which are phosphatidyl serine and phosphatidyl coline as major components and phosphatidyl inositol as minor component plus 3 phospholipids in trace proportion, from every cultivar at the early growth-stage.

• Journal of Microbiology and Biotechnology
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• v.18 no.11
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• pp.1862-1867
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• 2008

Streptochlorin is a small molecule isolated from marine Streptomyces sp. that is known to have antiangiogenic and anticancer properties. In this study, we examined the effects of this compound on reactive oxygen species (ROS) production and the association of these effects with apoptotic tumor cell death, using a human hepatocarcinoma Hep3B cell line. The results of this study demonstrated that streptochlorin mediates ROS production, and that this mediation is followed by a decrease in the mitochondrial membrane potential (MMP, ${\Delta}{\Psi}_m$), activation of caspase-3, and downregulation of antiapoptotic Bcl-2 protein. The quenching of ROS generation by N-acetyl-L-cysteine administration, a scavenger of ROS, reversed the streptochlorin-induced apoptosis effects via inhibition of ROS production, MMP collapse, and the subsequent activation of caspase-3. These observations clearly indicate that ROS are involved in the early molecular events in the streptochlorin-induced apoptotic pathway. Taken together, our data imply that streptochlorin-induced ROS is a key mediator of MMP collapse, which leads to the caspase-3 activation, culminating in apoptosis.

• Proceedings of the Korean Biophysical Society Conference
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• 2003.06a
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• pp.68-68
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• 2003

HP(2-20) (AKKVFKRLEKLEKLFSKIQNDK) derived from the N-terminus of Helicobacter pylori Ribosomal Protein L1 shows potent antimicrobial activity against bacterial, fungi and cancer cells without cytotoxic effect. In order to investigate the relationships between antimicrobial activity and the structures, several analogues have been designed and synthesized. The structures of these peptides in SDS micelles have been investigated using NMR spectroscopy and they revealed that analogue 3 has the longest, well-defined alpha-helix from Val5 to Trp19. NOESY experiments performed on HP and its analogues in nondeuterated SDS micelles show that protons in the indole ring of Trp16 are in close contact with methylene protons of SDS micelles. In order to probe the position of HP and its analogues relative to the SDS micelles, spin-labeled stearate was added. Large effects are observed for the chemical shifts and the intensities of Phe5, Glu9, Phe12, and Trp16 within the helix region by 16-doxylstearate. This result implies that 16-doxylstearate is located in the center of the micelles and the hydrophobic phase of the amphiphilic ${\alpha}$-helix is located in contact with the acyl chains of the micelles. Also, Lys3 and Lys4 at N-terminus and Lys20 at C-terminus may produce an optimal arrangement for electrostatic interactions between the sulfate head groups of the SDS and the positively charged lysyl N$\sub$3/$\^$+/. Interactions between the indole ring of Trp and the membrane, as well as the amphiphilic ${\alpha}$-helical structure of HP induced by Trp at the C-terminus may allow HP to span the lipid bilayer. These structural features are crucial for their potent antibiotic activities.

• Journal of the Korean Chemical Society
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• v.36 no.3
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• pp.393-404
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• 1992

The response properties of continuous automated system using an enzyme reactor for determination of urea were investigated. The enzyme reactor was constructed to packed-bed form which filled with nylon-6 beads (42∼48 mesh), which immobilized urease with glutaraldehyde, in teflon tube (2 mm I.D., 20 cm length). The system was composed of the enzyme reactor, gas dialyzer, and tublar PVC-nonactin membrane ammonium ion-selective electrode as an indicator electrode in serial order. The response characteristics of this system were as follows. That is, the concentration range of linear response, slope of linear response, detection limit, and conversion percentage were $5.5{\times}10^{-6}$∼$2.4{\times}10^{-3}M$, 57.8 mV/decade, $1.5{\times}10^{-6}$, and 80.8%, respectively. The optimum buffer and life time of urease reactor were 0.01M Tris-HCl buffer solution (pH 7.0∼7.8) and 0.01M phosphate buffer solution (pH 6.9∼7.5) and about 150 days, respectively. And the urease reactor had no interferences of the other physiological materials.

• Bulletin of the Korean Chemical Society
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• v.24 no.10
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• pp.1478-1484
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• 2003

CRAMP was identified from a cDNA clone derived from a mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptide. Tertiary structure of CRAMP in TFE/$H_2O$ (1 : 1, v/v) solution has been determined by NMR spectroscopy previously and consists of two amphipathic $\alpha-helices$ from Leu4 to Lys10 and from Gly16 to Leu33. These two helices are connected by a flexible region from Gly11 to Gly16. Analysis of series of fragments composed of various portion of CRAMP revealed that an 18-residue fragment with the sequence from Gly16 to Leu33 (CRAMP-18) was found to retain antibacterial activity without cytotoxicity. The effects of two Phe residues at positions 14 and 15 of CRAMP-18 on structure, antibacterial activity, and interaction with lipid membranes were investigated by $Phe^{14,15}$ ${\rightarrow}$ Ala substitution (CRAMP-18-A) in the present study. Substitution of Phe with Ala in CRAMP-18 caused a significant reduction on antibacterial and membrane-disrupting activities. Tertiary structures of CRAMP-18 in 50% TFE/$H_2O$ (1 : 1, v : v) solution shows amphipathic ${\alpha}$-helix, from $Glu^2{\;}to{\;}Leu^{18}$, while CRAMP-18-A has relatively short amphipathic ${\alpha}$-helix from $Leu^4{\;}to{\;}Ala^{15}$. These results suggest that the hydrophobic property of $Phe^{14}{\;}and{\;}Phe^15$ in CRAMP-18 is essential for its antibacterial activity, ${\alpha}$-helical structure, and interactions with phospholipid membranes.