• Molecules and Cells
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• v.39 no.6
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• pp.468-476
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• 2016

PLZF-expressing invariant natural killer T cells and CD4 T cells are unique subsets of innate T cells. Both are selected via thymocyte-thymocyte interaction, and they contribute to the generation of activated/memory-like CD4 and CD8 T cells in the thymus via the production of IL-4. Here, we investigated whether $PLZF^+$ innate T cells also affect the development and function of $Foxp3^+$ regulatory CD4 T cells. Flow cytometry analysis of the thymus and spleen from both CIITA transgenic C57BL/6 and wild-type BALB/c mice, which have abundant $PLZF^+$ CD4 T cells and invariant natural killer T cells, respectively, revealed that $Foxp3^+$ T cells in these mice exhibited a $CD103^+$ activated/memorylike phenotype. The frequency of $CD103^+$ regulatory T cells was considerably decreased in $PLZF^+$ cell-deficient $CIITA^{Tg}Plzf^{lu/lu}$ and $BALB/c.CD1d^{-/-}$ mice as well as in an IL-4-deficient background, such as in $CIITA^{Tg}IL-4^{-/-}$ and $BALB/c.IL-4^{-/-}$ mice, indicating that the acquisition of an activated/ memory-like phenotype was dependent on $PLZF^+$ innate T cells and IL-4. Using fetal thymic organ culture, we further demonstrated that IL-4 in concert with TGF-${\beta}$ enhanced the acquisition of the activated/memory-like phenotype of regulatory T cells. In functional aspects, the activated/ memory-like phenotype of Treg cells was directly related to their suppressive function; regulatory T cells of $CIITA^{Tg}PIV^{-/-}$ mice more efficiently suppressed ovalbumin-induced allergic airway inflammation compared with their counterparts from wild-type mice. All of these findings suggest that $PLZF^+$ innate T cells also augmented the generation of activated/memory-like regulation via IL-4 production.

• Molecules and Cells
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• v.40 no.8
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• pp.567-576
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• 2017

The $Na^+/H^+$ exchanger is responsible for maintaining the acidic tumor microenvironment through its promotion of the reabsorption of extracellular $Na^+$ and the extrusion of intracellular $H^+$. The resultant increase in the extracellular acidity contributes to the chemoresistance of malignant tumors. In this study, the chemosensitizing effects of cariporide, a potent $Na^+/H^+-exchange$ inhibitor, were evaluated in human malignant mesothelioma H-2452 cells preadapted with lactic acid. A higher basal level of phosphorylated (p)-AKT protein was found in the acid-tolerable H-2452AcT cells compared with their parental acid-sensitive H-2452 cells. When introduced in H-2452AcT cells with a concentration that shows only a slight toxicity in H-2452 cells, cariporide exhibited growth-suppressive and apoptosis-promoting activities, as demonstrated by an increase in the cells with pyknotic and fragmented nuclei, annexin V-PE(+) staining, a $sub-G_0/G_1$ peak, and a $G_2/M$ phase-transition delay in the cell cycle. Preceding these changes, a cariporide-induced p-AKT down-regulation, a p53 up-regulation, an ROS accumulation, and the depolarization of the mitochondrial-membrane potential were observed. A pretreatment with the phosphatidylinositol-3-kinase (PI3K) inhibitor LY294002 markedly augmented the DNA damage caused by the cariporide, as indicated by a much greater extent of comet tails and a tail moment with increased levels of the p-histone H2A.X, $p-ATM^{Ser1981}$, $p-ATR^{Ser428}$, $p-CHK1^{Ser345}$, and $p-CHK2^{Thr68}$, as well as a series of pro-apoptotic events. The data suggest that an inhibition of the PI3K/AKT signaling is necessary to enhance the cytotoxicity toward the acidtolerable H-2452AcT cells, and it underlines the significance of proton-pump targeting as a potential therapeutic strategy to overcome the acidic-microenvironment-associated chemotherapeutic resistance.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.419-424
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• 2009

To provide the basis of biosensor based on the lux genes from bioluminescent bacteria of Photobacterium leiognathi and Vibrio harveyi, we test the expression of lux genes in several strains of Escherichia coli. The expression of the recombinant plasmid of PlXba.pT7-3, containing all lux genes requiring for light emission without adding substrate, in E. coli 43R was so strong to see the blue-green light in single colony as well as in the alginate immobilized cell. In addition, the light intensity was decreased by adding heavy metal ion such as cadmium and zinc ions. These result raise the possibility that a biosensor can be developed using the lux genes system.

• Korean Journal of Microbiology
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• v.45 no.4
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• pp.324-331
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• 2009

MK1 strain, an obligate aerobic heterotrophic bacterium isolated from the rotten tissue of Neungee mushroom (Sarcodon aspratus), produces a copious amount of exopolysaccharide (EPS), which could evoke macrophage activation. Investigations on optimal culture conditions of MK1 and physical properties of MK1 EPS were made. Glucose, galactose, fructose, and sucrose supported well growth of MK1, but potato starch and dextrin did not. However, lactose seemed to be a less favorable carbon source. Optimal growth of MK1 was obtained at pH 7.0, $30^{\circ}C$, and 200 rpm with 2% glucose, and 0.2~0.05% $(NH_4)_2SO_4$. $EPS_{opt}$ obtained from an optimal growth condition constituted of carbon (37.1%), nitrogen (2.2%), oxygen (49.3%), and hydrogen (6.4%), but no sulfur. Paper chrogromatogram of the acid-hydrolysate of $EPS_{opt}$ suggested that MK1 EPS seemed to be hetropolysaccharide composed of a few number of monosaccharides including amino- and acidic-sugars. Its molecular mass determined by SDS-polyacrylamide gel electrophoresis varied from 14.8 to 47.9 kDa. Physical properties of $EPS_{glu}$ obtained from cell grown in glucose medium, such as relative viscosity ($_{rel}$) and crystalline morphology were rather affected by pH of the growth medium. Relative viscosity ($_{rel}$) of exopolysaccaride (0.1 g/ml) harvested from cells grown at medium pH ranging from 6.0 and 7.5 was 1.23 and 1.39, respectively. The freeze-dried exopolysaccharide obtained at low pH (6.0 and 6.5) was fine crystaloid and water-soluble, whereas those obtained at high pH (7.0 and 7.5) was rather gluey and less water-soluble.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.277-281
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• 2008

HIV affects many organ systems. Patients with HIV infection have substantially increased risk of developing various cancers, primarily by opportunistic infection with oncogenic viruses due to their immunocompromised status. However, extensive evidence also indicates that the viral protein, Tat itself, may playas a major factor in the development of AIDS-related neoplasms. The molecular mechanism underlying Tat's oncogenic activity may include deregulation of cellular genes. Therefore, in this study, we examined the effect of HIV-l Tat on CD99 as one of the target cellular genes, which is a well-known tumor marker in several cancers. By using established HeLa clones that are stably expressing Tat, we found that CD99 is upregulated by endogenous Tat, whereas STAT3 is down regulated. Upon the screening of genes differentially expressed between Tat-stable cells and the control cells by using the gene fishing technique, DEG, we detected 3 genes which expression is affected by the presence of Tat. Furthermore, the methylation specific PCR analysis of the stably Tat expressing cell lines revealed that the CD99 promoter is de methylated in the presence of Tat. Taken together, these results open a potential role of CD99 in AIDS-related oncogenesis via epigenetic regulation by HIV-1 Tat.

• Journal of the Korean Geotechnical Society
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• v.20 no.1
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• pp.109-120
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• 2004

There has been outstanding research on the soil-water characteristic curves of unsaturated soils over the past several decades. Unfortunately, unsaturated soil mechanics has not been considered as an important factor in Korea. In this paper, laboratory test and numerical analysis(SoilVision Professional ver 3.04) were performed to investigate the prediction method of soil-water characteristic curve and unsaturated permeability coefficient in reclaimed ground. The pressure cell, desiccator, and tensiometor tests were conducted on three types of reclaimed soils(dredged soil, sand, weathered granite soil). Numerical analysis was executed to compare the results with the laboratory test results and also compared with the results of each prediction method. Based on the laboratory test, three different types of soils have shown different soil-water characteristic curves. The hysteresis fir these soils is clearly defined. As a result of numerical analysis, Fredlund & Xing's method and Fredlund & Wilson's model proved to worke out well for reclaimed ground soils in Korea. Also, predicting method based on the soil-water characteristic curves from the particle-size distributions is flirty reliable for estimating unsaturated permeability coefficient.

• Journal of the Korean Geotechnical Society
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• v.28 no.9
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• pp.23-30
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• 2012

This paper is research results of slope stability analysis associated with seepage infiltration for unsaturated soil in debris-flow occurrence slopes. Site investigations were carried out in two slopes, located at Inje in Kangwon province where debris flow occurred in 2006 and at Yangpyung in Kyeunggi province where it occurred in 2010. For unsaturated soil sampled at the zone of debris-flow initiation, soil water characteristic curves with tempe pressure cells and shear strength parameters with newly designed shear strength apparatus were obtained respectively. The commertially available software SEEP/W was used to analyze seepage infiltration in unsaturated soil, based on their properties obtained from test results and the actual rainfall data at the moment of debris flow occurrence, and slope stability analysis with the program of SLOPE/W, associated with results of seepage analysis, was performed to simulate slope failure. As results of this research, seepage infiltration to unsaturated soil due to intensive rainfall was found to cause increase of ground water table as well as degree of saturation. Through this research slope stability analysis for unsaturated soil, considering the actual rainfall characteristic, might be a reasonable method of investigating characteristics of debris flow behavior, in particular, the moment of debris flow occurrence.

• Journal of the Korean Geotechnical Society
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• v.28 no.12
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• pp.41-51
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• 2012

Soils are commonly unsaturated in the near surface. The stiffness of soils is affected by the amount of air and water. The objective of this study is to evaluate the porosity of the unsaturated soils by using the elastic waves including compressional and shear waves. The elastic waves are measured at different degrees of saturation by controlling the matric suction. Thus, the unsaturated soils are characterized at different levels of the matric suction. Shear and compressional waves are measured by using the bender elements and the piezo disk elements, respectively. Both transducers are installed on the walls of the rectangular cell. The unsaturated soils are prepared by using uniform size sands and silts. Test results show that both compressional and shear wave velocities change according to the matric suction. The elastic modulus, the shear modulus, and the Poisson's ratio are estimated based on the measured elastic wave velocities. In addition, the void ratio of the unsaturated soils estimated using elastic wave velocities matches well with the volume based void ratio. This study demonstrates that the elastic waves can be effectively used for the characterization of unsaturated soils.

• KSBB Journal
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• v.21 no.2
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• pp.133-139
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• 2006

In 'solid-phase' PEGylation, the conjugation reaction occurs as the proteins are attached to a solid matrix, and thus it can have distinct advantages over the conventional, solution-phase process. We report a case study: rhIFN-${\alpha}$-2a was first adsorbed to cation exchange resin and then N-terminally PEGylated by aldehyde mPEG of 5, 10, and 20 kD through reductive alkylation. After the PEGylation, salt gradient elution efficiently recovered the mono-PEGylate in a purified form from the unwanted species such as unmodified IFN, unreacted PEG, and others. The mono-PEGylation and its purification were integrated in a single chromatographic step. Depending on the molecular weight of the mPEG aldehyde used, the mono-PEGylation yield ranged 50-64%. We could overcome the major problems of random, or uncontrollable, multi-PEGylation and the post-PEGylation purification difficulties associated with the solution-phase process. N-terminal sequencing and MALDI-TOF MS confirmed that a PEG molecule was conjugated only to the N-terminus. Compared with the unmodified IFN, the mono-PEGylate showed the reduced anti-viral activity as measured by the cell proliferation assay. The bioactivity was reduced more as the higher molecular weight PEG was conjugated. Immunoreactivity, evaluated indirectly by antibody binding activity using a surface plasmon resonance biosensor, also decreased. Nevertheless, trypsin resistance as well as thermal stability was considerably improved.

• Korean Chemical Engineering Research
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• v.45 no.3
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• pp.242-248
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• 2007

For an optimal design of automotive electric system, it is important to have a reliable modeling tool to predict the charge-discharge behaviors of the automotive battery. In this work, a two-dimensional modeling was carried out to predict the charge-discharge behaviors of a 12-V automotive lead-acid battery. The model accounted for electrochemical kinetics and ionic mass transfer in a battery cell. In order to validate the modeling, modeling results were compared with the experimental data of the charge-discharge behaviors of a lead-acid battery. The discharge behaviors were measured with three different discharge rates of C/5, C/10, and C/20 at operating temperature of $25^{\circ}C$. The batteries were charged with constant current of 30A until the charging voltage reached to a predetermined value of 14.24 V and then the charging voltage was kept constant. The discharge and charge curves from the measurements and modeling were in good agreement. Based on the modeling, the distributions of the electrical potentials of the solid and solution phases, the porosity of the electrodes, and the current density within the electrodes as well as the acid concentration can be predicted as a function of charge and discharge time.