• BMB Reports
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• 제39권2호
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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.

• 한국식품영양과학회지
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• 제25권6호
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• pp.922-927
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• 1996

본 연구에서는 열처리에 의한 $알파-락트알부민(\alpha-La)겔의$ 특성을 규명하기 위하여 겔형성에 관여하는 인자들 즉, 염의 종류와 농도, pH, $\alpha-La$ 농도, 티올시약(NEM, Dn)의 농도를 각각 변화시켜 $90^{\circ}C에서$ 40분간 가열하여 만든 알파-락트알부민 겔의 총유리 SH그룹, half-cystine 함량, S-S 결합 함량을 측정하였다. 총 유리 SH 그룹, half-cystine 함량, S-S 결합은 첨가된 염의 종류와 농도 변화에는 큰 의존성을 나타내지 않았다. pH 2.5~3.5에서는 SH 그룹의 반응성이 낮아 pH 6.5~8.5에서의 총 유리 SH 그룹 보다 함량이 더 높고 half-cystine 함량은 일정하였으나 형성된 S-S 결합은 더 낮아 pH가 증가할수록 SH산화속도와 겔망상 구조의 형성 이 가속화됨을 확인할 수 있었다. $\alpha-La의$ 농도 증가로 half-cystine 함량은 큰 변화가 없었으나 총 유리 SH 그룹은 약간 감소하고 S-S 결합은 약간 증가하여 $\alpha-La의$ 농도 증가에 따라 겔지지체에서 S-S 결합의 관여가 큼을 나타내었다. NEM첨가의 경우는 총 유리 SH그룹과 half-cystine 함량이 급격히 감소하였으나 S-S 결합은 증가하여 NEM 첨가로 SH그룹이 반응성을 잃어 결국 $\alpha-La의$ 겔형성에 지장을 줌을 확인하였다. DTT 첨가로는 새로운 분자간 티올-이황화물 상호교환반응이 용이하게 되어 총 유리 SH 그룹은 낮고 S-S 결합은 높은 함량을 나타내었다.

• 한국환경보건학회지
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• 제27권2호
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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.

• 대한방사선기술학회지:방사선기술과학
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• 제43권6호
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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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• 제34권2호
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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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• 제38권6호
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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.

• 한국미생물·생명공학회지
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• 제30권4호
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• pp.320-324
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• 2002

우리나라 연안에 주로 발생되는 적조인 Cochlodinium polykrikoides를 빠르고 정밀한 생화학적인 방법으로 측정하기 위한 일종의 maker로서 C. polykrikoides의 세포막에 존재하는 특이 항원성을 가진 막 단백질을 분리하였다. 먼저, C. polykrikoides와 gymnodium sangineum의 세포를 삼투압으로 터뜨린 후 원심분리하여 세포막을 모았다. 그 후 양 적조생물의 세포막은 1 mM DTT가 함유된 50 mM Na-carbonate로 용해하고 SDS-PAGE행하여 용해된 막 단백질을 분리하였다. SDS-PAGE로 분리된 막 단백질은 C. polykrikoides의 세포 막 단백질로 제조한 항혈청을 사용하여 immune-blot한 결과 C. polykrikoides의 120 kDa의 단백질이 G. sangineum의 동일한 크기의 막 단백질과는 서로 다른 항원성을 나타내었다.

• 한국광학회:학술대회논문집
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• 한국광학회 2000년도 하계학술발표회
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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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• 제14권1호
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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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• 제24권3호
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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.