• BMB Reports
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• v.39 no.2
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• pp.216-222
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• 2006

In the present study, we compared six different solubilization buffers and optimized two-dimensional electrophoresis (2-DE) conditions for human lymph node proteins. In addition, we developed a simple protocol for 2-D gel storage. Efficient solubilization was obtained with lysis buffers containing (a) 8M urea, 4% CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate), 40 mM Tris base, 65 mM DTT(dithiothreitol) and 0.2% carrier ampholytes; (b) 5M urea, 2M thiourea, 2% CHAPS, 2% SB 3-10 (N-decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate), 40mM Tris base, 65 mM DTT and 0.2% carrier ampholytes or (c) 7M urea, 2M thiourea, 4% CHAPS, 65 mM DTT and 0.2% carrier ampholytes. The optimal protocol for isoelectric focusing (IEF) was accumulated voltage of 16,500 Vh and 0.6% DTT in the rehydration solution. In the experiments conducted for the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), best results were obtained with a doubled concentration (50 mM Tris, 384 mM glycine, 0.2% SDS) of the SDS electrophoresis buffer in the cathodic reservoir as compared to the concentration in the anodic reservoir (25 mM Tris, 192 mM glycine, 0.1% SDS). Among the five protocols tested for gel storing, success was attained when the gels were stored in plastic bags with 50% glycerol. This is the first report describing the successful solubilization and 2D-electrophoresis of proteins from human lymph node tissue and a 2-D gel storage protocol for easy gel handling before mass spectrometry (MS) analysis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.6
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• pp.922-927
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• 1996

In order to investigate heat-induced gelation properties of $\alpha-lactalbumin(\alpha-La),$ total free SH groups, half-cystine, and disulfide bond contents of $\alpha-La$ gels prepared in 0.1M Tris-HCI buffer(pH 8.0) were measured. The samples were heated at $90^{\circ}C$ for 40 minutes under different PH and concentrations of NaCl, $CaCl_2,$ $\alpha-La,$ N-ethylrnaleirnide(NEM), and dithiothreitol(DTT). Total free SH groups were low at high concentrations of $\alpha-La$ and at pH 6.5~8.5, and were $14.72~18.58\mu\textrm{m}ol/g$ and $14.17~16.11\mu\textrm{m}ol/g,$ respectively. Half-cystine contents of NEM-induced gels decreased with increasing concentration of NEM, and were $1.03~39.17\mu\textrm{m}ol/g.$ Disulfide bonds of DTT-induced gels increased with increasing concentration of DTT, and were $70.04~71.80\mu\textrm{m}ol/g$.

• Journal of Environmental Health Sciences
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• v.27 no.2
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• pp.82-91
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• 2001

This study was carried out to elucidate the protective effect of zinc chloride(ZnCl$_2$) and its mechanism against the immuno-cytotoxicity of methylmercury chloide($CH_3$HgCl). This study was observed in the culture of EMT-6 cells which are originated from mammary adenocarcinoma of Balb/c mouse. Cytotoxicity of metals was measured by cell viability and NO$_2$$^{[-10]}$ , and mitochondrial function was evaluated by adenosine triphosohate (ATP) production. $CH_3$HgCl significantly decreased the sythesis of nitric oxide(NO), ATP and glutathione(GSH) in a dose-dependent manner. ZnCl$_2$ significantly increased the synthesis of GSH in a dose-dependent manner, but synthesis of NO and ATP were not changed. The immuno-cytotoxicity of $CH_3$HgCl was not fully protected when combined addition of ZnCl$_2$, whereas ZnCl$_2$ prior to addition of $CH_3$HgCl completly protected the Hg-induced immuno-cytotoxicity. Similarly, intracellular accumulation of mercury significantly decreased by ZnCl$_2$. Degree of diminution of intracellular mercury was larger in ZnCl$_2$ prior to addition of $CH_3$HgCl than in combined addition of ZnCl$_2$ and $CH_3$HgCl.. Dithiothreitol(DTT) or buthionine sulfoximine(BSO) addition at 50$\mu$M or less, which was not toxic to the cells, did not affect synthesis of NO and ATP. DTT increased intracellular GSH level and DTT pretreatment protected toxicity induced by $CH_3$HgCl as shown complete recover in the NO and ATP values. BSO decreased intracellular GSH level and BSO pretreatment exaggerated toxicity induced by $CH_3$HgCl as shown synergistic reduction in the NO and ATP values. These results indicated that the protective effects of zinc against immuno-cytotoxicity of methylmercury associated with increasing cellular level of GSH. Increased intracellular GSH transports methylmercury to out of cells. In accordance with intracellular level of mercury decreased, immuno-cytotoxicity of methylmercury decreased. These result also suggest that the protective mechanism of zinc against the mercury toxicity would be exerted in the immune system in vivo.

• Journal of radiological science and technology
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• v.43 no.6
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• pp.475-480
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• 2020

The purpose of this study was to reconstruct diffusion tensor tractography (DTT) using stem of garlic and asparagus for in vitro phantom of diffusion tensor imaging (DTI), and to compare and evaluate the fractional anisotropy (FA) value and the apparent diffusion coefficient (ADC) value to determine whether it can be used as materials for in vitro phantoms. Among various plant fibers such as stem of garlic, palmae, cotton, asparagus, etc., stem of garlic and asparagus, which are considered to be the most suitable for making phantoms, and whose shape is considered to be the most suitable for making phantoms, were selected and tests were conducted. Holes were made in a plastic bucket at an angle of 0°, 30°, 60°, 90°, and 120°, then tubes were inserted. In the tube, asparagus and stem of garlic were inserted as far in as possible, and the inserted tube was inserted into the center of the heat bathed gelatin to harden. We were able to reproduce DTT images in asparagus and stem of garlic. Fiber tissues of asparagus and stem of garlic did not show complete connectivity, but the reconstructed images of DTT showed good connectivity. The FA values of asparagus in the tubes were 0.198 at 0° (straight), 0.207 at 30°, 0.187 at 60°, 0.231 at 90°, and 0.204 at 120°. In addition, the FA values of stem of garlic in the tubes were 0.235 at 0°, 0.236 at 30°, 0.216 at 60°, 0.218 at 90°, and 0.257 at 120°. The ADC values of asparagus in the tubes were 1.545 at 0°, 1.677 at 30°, 1.629 at 60°, 1.535 at 90°, and 1.725 at 120°. In addition, the ADC values of stem of garlic in the tubes were 1.252 at 0°, 1.396 at 30°, 1.698 at 60°, 1.756 at 90°, and 1.466 at 120°. For the best expressed DTT reconstruction image, it showed the longest connectivity in the straight line as we hypothesized. In addition, when comparing the FA values and ADC values of fiber tissues of stem of garlic and asparagus, FA value was generally higher in stem of garlic and ADC value was slightly higher in asparagus.

• BMB Reports
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• v.34 no.2
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• pp.102-108
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• 2001

A time-dependent folding process was used to determine whether or not protein disulfide isomerase (PDI) plays an important role in the maturation of nascent lactoferrin polypeptides. Interaction between lactoferrin and PDI was analyzed according to the co-immunoprecipitation of the two proteins. The results indicate that lactoferrin folding requires a significant interaction with PDI and its binding is relatively brief compared to other nascent polypeptides. The amount of lactoferrin interacting with PDI increases up to half a minute and sharply decreases beyond this time point. During the refolding process that follows reduction by DTT, lactoferrin polypeptides heavily interact with PDI and the interaction period was extended compared to the normal folding process. In terms of the temperature effect on PDI-lactoferrin interaction, PDI binds to lactoferrin polypeptides longer at a lower temperature (here, $25^{\circ}C$) than $37^{\circ}C$. The lactoferrin-PDI interaction was also studied in vitro. According to the in vitro experiment data, PDI was still functional in cell lysates assisting lactoferrin folding into the mature form. PDI interacts with lactoferrin polypeptides for an extended period during the folding in vitro. During the refolding process in vitro, intermolecular aggregates and refolding oligomers matured into a functional form after PDI binds to the lactoferrin. These results suggest that PDI provides a prolonged chaperoning activity in the refolding processes and that there appears to be a greater requirement for PDI chaperone activity in the refolding of lactoferrin polypeptides.

• BMB Reports
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• v.38 no.6
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• pp.763-765
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• 2005

A new simplified procedure for identifying human plasmin was developed using a DTT copolymerized agarose stacking gel (ASG) system. Agarose (1%) was used for the stacking gel because DTT inhibits the polymerization of acrylamide. Human plasmin showed the lowest activity at pH 9.0. There was a similar catalytically active pattern observed under acidic conditions (pH 3.0) to that observed under alkaline conditions (pH 10.0 or 11.0). Using the ASG system, the primary structure of the heavy chain could be established at pH 3.0. This protein was found to consist of three fragments, 45 kDa, 23 kDa, and 13 kDa. These results showed that the heavy chain has a similar structure to the autolysed plasmin (Wu et al., 1987b) but there is a different start amino acid sequence of the N-termini.

• Microbiology and Biotechnology Letters
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• v.30 no.4
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• pp.320-324
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• 2002

To establish a rapidly immunochemical identification on a dinoflagellate, Cochlodinium polykrikoides, a specific antigenic protein as a maker on the cell membrane was isolated. The cell membranes of C. polykrikoides and Gymnodinium sangineum were harvested by centrifugation after osmotic shock. The membrane proteins of both cells were solubilized in 50 mM Na-carbonate contained 1 mM DTT, and separated the proteins on SDS-PACE. Immune-blot on the solubilized membrane proteins of the both cells was performed with antiserum against the solubilized membrane proteins of C. polykrikoides. A 120 kDa membrane protein of C. polykrikoides had remarkablely different antigenicity from that of G. sangineum.

• Proceedings of the Optical Society of Korea Conference
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• 2000.08a
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• pp.164-165
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• 2000

There has recently been a considerable amount of interest in two-photon(TP) processes because of their applications to 3-D optical data storage and imaging, up-conversion lasing, and optical power limiting. Recently, the role of Dithienothlophene(DTT) as $\pi$-center of two-photon absorption(TPA) molecules was reported to give a large enhancement of TPA, compared with the benzeniods counterparts.$^{[1]}$ In this report, we carried out two experiments to investigate nonlinear optical absorption of the molecules in which central DTT is attached through conjugation to either a D/D or a D/A pair at the ends, forming a D-$\pi$-D(1 and 3) or D-$\pi$-A(2 and 4) sequence(Fig.1). All the samples were prepared in solutions with the same molar concentration. (omitted)

• PNF and Movement
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• v.14 no.1
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• pp.41-47
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• 2016

Purpose: In general, confabulation is defined as confusion of reality with past events without apparent prompting, in association with disruption of the capacity for retrieval and encoding of memory. We report on a patient who showed spontaneous confabulation associated with injury of the Papez circuit following middle cerebral artery (MCA) infarction. Methods: A 67-year-old female patient suffered cerebral infarct resulting from spontaneous MCA territory. After onset of the MCA infarct, she showed severe memory impairment and provoked confabulation. The Papez circuit was reconstructed for evaluation of part of it using diffusion tensor tractography (DTT). Fractional anisotropy (FA), mean diffusivity (MD), and tract volume were measured. Results: The right thalamocingulate tract showed a significant decrement of FA value and tract volume, and an increment of MD value by more than two standard deviations of that of normal control subjects. The tract volume in the left fornix and mammillothalamic tract decreased by more than two standard deviations of that of normal control subjects. Conclusion: Injuries of the Papez circuit were demonstrated in a patient who showed severe memory impairment and provoked confabulation following MCA infarct. We believe that analysis of the Papez circuit tract using DTT is useful in elucidating the cause of provoked confabulation in patients with MCA infarct.

• ETRI Journal
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• v.24 no.3
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• pp.221-225
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• 2002

We performed nonlinear transmission measurements and quantum-chemical calculations on dithienothiophene(DTT)-based molecules to gain insight into the effect of acceptor and donor groups on two-photon absorption(TPA) properties. The TPA intensity showed dispersion characteristics of the single-photon absorption spectrum. When the molecules included an asymmetric donor-acceptor pair, the single- and two-photon absorption maximum wavelengths were red-shifted more than when the molecules had a symmetric donor-donor structure. We interpreted this result as indicating that the $S_2$ state plays the dominating role in the absorption process of molecules with a symmetric structure. The experimental TPA ${\delta}$ values at the absorption peak wavelength showed a dependence on the structural variations. We found the self-consistent force-field theory and Hartree-Fock Hamiltonian with single configuration interaction formalism to be valid for evaluating TPA ${\delta}$. Although the quantum-chemical calculations slightly underestimated the experimental ${\delta}$ values obtained from nonlinear trans -mission measurements, they reasonably predicted the dependence of the ${\delta}$ value on the structural variations. We confirmed the role of molecular symmetry by observing that donor-donor substituted structure gave the highest experimental and theoretical TPA ${\delta}$ values and that the donor-acceptor substituted structure showed a greater red-shift in the TPA absorption maximum wavelength. Overall, the theoretical ${\delta}$ values of DTT-based molecules were in the order of $10^{-46}\;cm^4{\cdot}s{\cdot}photon^{-1}$ and are higher than that of AF-50 by nearly two orders of magnitude.