• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1323-1329
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• 2021

Micro-scale magnetic beads are widely used for isolation of proteins, DNA, and cells, leading to the development of in vitro diagnostics. Efficient isolation of target biomolecules is one of the keys to developing a simple and rapid point-of-care diagnostic. A zinc finger protein (ZFP) is a double-stranded (ds) DNA-binding domain, providing a useful scaffold for direct reading of the sequence information. Here, we utilized two engineered ZFPs (Stx2-268 and SEB-435) to detect the Shiga toxin (stx2) gene and the staphylococcal enterotoxin B (seb) gene present in foodborne pathogens, Escherichia coli O157 and Staphylococcus aureus, respectively. Engineered ZFPs are immobilized on a paramagnetic bead as a detection platform to efficiently isolate the target dsDNA-ZFP bound complex. The small paramagnetic beads provide a high surface area to volume ratio, allowing more ZFPs to be immobilized on the beads, which leads to increased target DNA detection. The fluorescence signal was measured upon ZFP binding to fluorophore-labeled target dsDNA. In this study, our system provided a detection limit of ≤ 60 fmol and demonstrated high specificity with multiplexing capability, suggesting a potential for development into a simple and reliable diagnostic for detecting multiple pathogens without target amplification.

• Journal of the Korean Magnetic Resonance Society
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• v.25 no.2
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• pp.17-23
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• 2021

High mobility group protein B1 (HMGB1) is a highly conserved, non-histone, chromatin associated nuclear protein encoded by HMGB1 gene. HMGB1 proteins may be general co-factors in Hox-mediated transcriptional activation that facilitate the access of Hox proteins to specific DNA targets. It is unclear that the exact binding interface of Hoxc9DBD and HMGB1. To identify the interface and binding affinity of Hoxc9DBD and HMGB1 A-box, the paramagnetic probe, MTSL was used in NMR titration experiment. It is attached to the N-terminal end of HMGB1 A-box by reaction with thiol groups. The backbone assignment of HMGB1 A-box was achieved with 3D NMR techinques. The ¹⁵N-labeled HMGB1 A-box was titrated with MTSL-labeled Hoxc9DBD respectively. Based on the chemical shift changes we can identify the interacting residues and further map out the binding sites on the protein structure. The NMR titration result showed that the binding interface of HMGB1 A-box is around loop-1 between helix-1 and helix-2. In addition, the additional contacts were found in N- and C-terminus. The N-terminal arm region of Hoxc9DBD is the major binding region and the loop between helix1 and helix2 is the minor binding region.

• Nuclear Engineering and Technology
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• v.55 no.1
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• pp.270-277
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• 2023

Exposure to ionizing radiation induces free radicals in human nails. These free radicals generate a radiation-induced signal (RIS) in electron paramagnetic resonance (EPR) spectroscopy. Compared with the RIS of tooth enamel samples, that in human nails is more affected by moisture and heat, but has the advantages of being sensitive to radiation and easy to collect. The fingernail as a biological sample is applicable in retrospective dosimetry in cases of localized hand exposure accidents. In this study, the dosimetric characteristics of fingernails were analyzed in fingernail clippings collected from Korean donors. The dose response, fading of radiation-induced and mechanically induced signals, treatment method for evaluation of background signal, minimum detectable dose, and minimum detectable mass were investigated to propose a fingernail-EPR dosimetry protocol. In addition, to validate the practicality of the protocol, blind and field experiments were performed in the laboratory and a non-destructive testing facility. The relative biases in the dose assessment result of the blind and field experiments were 8.43% and 21.68% on average between the reference and reconstructed doses. The results of this study suggest that fingernail-EPR dosimetry can be a useful method for the application of retrospective dosimetry in cases of radiological accidents.

• Journal of the Korean Chemical Society
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• v.25 no.6
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• pp.367-375
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• 1981

Single crystals of Potassium Sulfate ($K_2SO_4$) have been grown from the saturated solution by the evaporation method at the optimum conditions. Radiation damages in the crystal by ${\gamma}$-irradiation of about $12{\times}10^6$ Roentgen have given rise to paramagnetic centers or paramagnetic defects. Electron spin resonance (ESR) spectra of the centers are obtained with the X-band EPR spectrometer at room temperature. The ESR peaks of the paramagnetic species are found to be anisotropic but the peak of $SO_3-$ radical is an isotropic of Gaussian shape at g = 2.0036. A number of ESR spectra of the crystal for angular variation of the anisotropic peaks are recorded at various orientations of rotation about a, b and c crystallographic axes respectively. The g-values are calculated from the line position between anisotropic peaks and the isotropic one and then principal g-values and its direction cosines of the species are obtained by diagonalization of 9 matrix elements of the corresponding g-values. All the paramagnetic defects are identified by the characteristic principal g-values and its direction cosines.

• Journal of the Korean Magnetics Society
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• v.24 no.3
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• pp.71-75
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• 2014

Ground state energy levels of the $Fe^{3+}$ paramagnetic impurity ion in stoichiometric $LiTaO_3$ and in congruent $LiTaO_3$ single crystals were calculated with electron paramagnetic resonance constants. Energy levels between six energy levels were obtained with spectroscopic splitting parameter g and zero field splitting constant D for $Fe^{3+}$ ion. The energy diagrams of $Fe^{3+}$ ion were different from different magnetic field directions ([100], [001], [111]) when magnetic field increases. The calculated ZFS energies of $Fe^{3+}$ ion in stoichiometric and congruent $LiTaO_3$ single crystals for ${\mid}{\pm}5/2$ > ${\leftrightarrow}{\mid}{\pm}3/2$ > and ${\mid}{\pm}3/2$ > ${\leftrightarrow}{\mid}{\pm}1/2$ > transitions were 12.300 GHz and 6.150 GHz, and 59.358 GHz and 29.679 GHz, respectively. It turns out that energy levels of $Fe^{3+}$ paramagnetic impurity in $LiTaO_3$ crystal are different from different crystal growing condition.

• Progress in Medical Physics
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• v.17 no.2
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• pp.61-66
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• 2006

To evaluate the T1, T2 magnetic relaxation properties of water molecule according to molecular weight of paramagnetic complex. 4-aminomethyicyclohexane carboxylic acid (0.63 g, 4 mmol) was mixed with the suspension solution of DMF (15 ml) and DTPA-bis-anhydride (0.71 g, 2 mmol) to synthesize the ligand. The ligand was then mixed with $Gd_2O_3$ (0.18 g, 0.5 mmol) to synthesize Gd-chelate. For the measurement of magnetic relaxivity of paramagnetic compounds, the compounds were diluted to 1 mM and then the relaxation times were measured at 1.57 (64 MHz). Inversion-recovery pulse sequence was employed for T1 relaxation measurement and CPMG (Carr-Purcell-Meiboon-Gill) pulse sequence was employed for T2 relaxation measurement. In case of inversion recovery sequence, total 35 images with different inversion time(T1)s ranging from 50 msec to 1,750 msec. To estimate the relaxation times, the signal intensity of each sample was measured using region of Interest (ROI) and then fitted by non-linear least square method to yield T1, T2 relaxation times and also R1 and R2. Compared to T1=($205.1{\pm}2.57$) msec and T2=($209.4{\pm}4.28$) msec of Omniscan (Gadodiamide), which is commercially available paramagnetic MR agent, T1 and T2 values of new paramagnetic complexes were reduced along with their molecular weight. That is, T1 value was ranged from $(96.35{\pm}2.04)\;to\;(79.38{\pm}1.55)$ msec and T2 value was ranged from $(91.02{\pm}2.08)\;to\;(76.66{\pm}1.84)$ msec. Among new paramagnetic complexes, there is a tendency that the R1 and R2 increase as the molecular weight is increases. As molecular weight of paramagnetic complex increases, T1 and T2 relaxation times reduce and thus the increase of relaxivity (R1 and R2) Is proportional to molecular weight.

• Proceedings of the KSMRM Conference
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• 2000.11a
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• pp.21-21
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• 2000

• Bulletin of the Korean Chemical Society
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• v.21 no.2
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• pp.251-258
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• 2000

• Korean Journal of Materials Research
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• v.14 no.9
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• pp.659-663
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• 2004

Magnetite was prepared by wet method and oil-based magnetic fluid also was prepared by its magnetite. Ahead of making magnetite, we investigated the variation of physical properties for changing $Fe^{3+}/Fe^{2+}$. Through the variation of the process mixing time, pH, magnetite content, we made magnetic fluid of magnetite. We examined the magnetic and frequency property by a toroidal mold. In result $Fe^{3+}/Fe^{2+}=1.2$, over the minute mixing time property of magnetite, Ms was 61.98 emu/g. Moreover, the B-H loop result indicated that the magnetic fluid followed paramagnetic behavior.

• Journal of the Korean Magnetic Resonance Society
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• v.18 no.1
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• pp.1-4
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• 2014

It is generally understood that protein-protein interactions proceed via transient encounter complexes that rapidly evolve into the functional stereospecific complex. Direct detection and characterization of the encounter complexes, however, been difficult due to their low population and short lifetimes. Recent application of NMR paramagnetic relaxation enhancement first visualized the structures of the encounter complex ensemble, and allowed the characterization of their physicochemical properties. Further, rational protein mutations that perturbed the encounter complex formation provided a clue to the target search pathway during protein-protein association. Understanding the structure and dynamics of encounter complexes will provide useful information on the mechanism of protein association.