• The Journal of the Korean Society for Microbiology
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• v.34 no.6
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• pp.501-512
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• 1999

Gliotoxin, a fungal metabolite, is one of the epipolythiodioxopiperazine classes and has a variety of effects including immunomodulatory and apoptotic agents. This study is designed to evaluate the effect of zinc on gliotoxin-induced death of HL-60 cells. Here, we demonstrated that treatment of gliotoxin decreased cell viability in a dose and time-dependent manner. Gliotoxin-induced cell death was confirmed as apoptosis characterized by chromatin margination, fragmentation and ladder-pattern digestion of genomic DNA. Gliotoxin increased the proteolytic activities of caspase 3, 6, 8, and 9. Caspase-3 activation was further confirmed by the degradation of procaspase-3 and PARP in gliotoxin-treated HL-60 cells. Zinc compounds including $ZnCl_2$ and $ZnSO_4$ markedly inhibited gliotoxin-induced apoptosis in HL-60 cells (from 30% to 90%). Consistent with anti-apoptotic effects, zinc also suppressed the enzymatic activities of caspase-3 and -9 proteases. In addition, cleavage of both PARP and procaspase 3 in gliotoxin-treated HL-60 cells was inhibited by the addition of zinc compounds. We further demonstrated that expression of Fas ligand by gliotoxin was suppressed by zinc compounds. These data suggest that zinc may prevent gliotoxin-induced apoptosis via inhibition of Fas ligand expression as well as suppression of caspase family cysteine proteases-3 and -9 in HL-60 cells.

• Biomolecules & Therapeutics
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• v.10 no.2
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• pp.71-77
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• 2002

Heme oxygenase-1 (HO-1) is the inducible from of the rate-limiting enzyme of heme degradation; it regulates the cellular contents of heme. HO-1 is up-regulated by various stimuli including oxidative stress so that it is thought to participate in general cellular defense mechanisms against oxidative stress in mammalian cells. To investigate the role of the cAMP-dependent protein kinase A (PKA) signaling pathway on nitrogen oxidative stress-induced HO-1 gene expression, RAW 264.7 cell cultures were treated with sodium nitroprusside (SNP). SNP increased the expression of HO-1 mRNA and protein, time- and concentration-dependently. Treatment with H89, PKA inhibitor, but not LY83583, guanylate cyclase inhibitor, significantly diminished the HO-1 expression by SNP, indicating that cAMP plays a crucial role in the induction of HO-1. Incubation with cAMP-elevating agents, such as forskolin or isoproterenol resulted in up-regulation of the expression of HO-1. Forskolin-induced expression of HO-1 was inhibited by H89. Furthermore, propranolol, $\beta$-adrenoceptor blocker, inhibited the isoproterenol-induced HO-1 expression, supporting the importance of cAMP in the induction of HO-1 expression. Higenamine-S, but not higenamineR, enhanced the HO-1 expression induced by SNP. Furthermore, cellular toxicity induced by hydrogen peroxide was attenuated by the presence of SNP, which was further increased by the presence of ZnPPIX, HO-1 inhibitor. Collectively, these results strongly suggest that up-regulation of HO-1 expression in RAW 264.7 cells involves PKA signal pathway.

• Animal cells and systems
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• v.4 no.2
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• pp.157-163
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• 2000

Yeast pheromone a-factor is a 13-amino acid peptide hormone that is synthesized as a part of a larger precursor, prepro-$\alpha$-factor, consisting of a signal peptide and a proregion of 64 amino acids. The carboxy-terminal half of the precursor contains four tandem copies of mature $\alpha$-factor. To investigate the molecular basis of intracellular sorting, proteolytic processing, and storage of the peptide hormone, yeast prepro-$\alpha$-factor precursors were heterologously expressed in rat pituitary $GH_3 cells. When cells harboring the precursor were metabolically labeled, a species of approximately 27 kD appeared inside the cells. Digestion with peptide: N-glycosidase F (PNG-F) shifted the molecular mass to a 19 kD, suggesting that the 27 kD protein was the glycosylated form as in yeast cells. The nascent polypeptide is efficiently targeted to the ER in the $GH_3 cells, where it undergoes cleavage of its signal peptide and core glycosylation to generate glycosylated pro-a-factor. To look at the post ER intracellular processing, the pulse-labelled cells were chased up to 2 hrs. The nascent propeptides disappeared from the cells at a half life of 30 min and only 10-25% of the newly synthesized, unprocessed precursors were stored intracellularly after the 2 h chase. However, about 20% of the pulse-labeled pro-$\alpha$-factor precursors were secreted into the medium in the pro-hormone form. With increasing chase time, the intracellular level of propeptide decreased, but the amount of secreted propeptide could not account for the disappearance of intracellular propeptide completely. This disappearance was insensitive to lysosomotropic agents, but was inhibited at $16^{circ}C or 20^{\circ}C$, suggesting that the turnover of the precursors was not occurring in the secretory pathway to trans Golgi network (TGN) or dependent on acidic compartments. From these results, it is concluded that a pan of these heterologous precursors may be processed at its paired dibasic sites by prohormone processing enzymes located in TGN/secretpry vesicles producing small peptides, and that the residual unprocessed precursors may be secreted into the medium rather than degraded intracellularly.

• Journal of Korean Society of Forest Science
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• v.98 no.5
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• pp.548-557
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• 2009

For centuries, forests have been a key component of rural livelihood. They are important both socially and economically in Nepal. Firewood and fodder are the basic forest products that are extracted daily or weekly basis in most of the rural areas in Nepal. In this study, a field survey of 100 households was conducted to examine the degree of forest dependency and forest resource availability, households' livelihood strategy and their relationship with forest dependency in Chitwan, Nepal. A household' response indexes were constructed, Gini coefficient, Head Count Poverty Index (HCI) and Poverty Gap Index (PGI) were calculated and one way ANOVA test was also performed for data analysis. Data revealed that 82/81% of all households were constantly used forest for firewood and fodder collection respectively while 42% of households were used forest or forest fringe for grazing. The Forest Product Availability Indexes (FPAI) showed a sharp decline of forest resources from 0.781 to 0.308 for a 20-yr time horizon while timber wood was noticeably lowered than the other products. Yet, about 33% of households were below the poverty threshold line with 0.0945 PGI. Income distribution among the household showed a lower Gini coefficient 0.25 than 0.37 of landholdings size. However, mean income was significantly varies with F-statistics=246.348 at P=0.05 between income groups (rich, medium and poor). The extraction of firewood, fodder and other forest products were significantly different between the income group with F-statistics=16.480, 19.930, 29.956 at P=0.05 respectively. Similarly, landholdings size and education were also significantly different between the income groups with F-statistics=4.333, 5.981 at P=0.05 respectively. These findings suggested that income status of households was the major indicator of forest dependency while poor and medium groups were highly dependent on the forests for firewood, fodder and other products. Forest dependency still remains high and the availability of forest products that can be extracted from the remaining forestlands is decreasing. The high dependency of households on forest coupled with other socioeconomic attributes like education, poverty, small landholders and so on were possibly caused the forest degradation in Chitwan.Therefore, policy must be directed towards the poor livelihood supporting agenda that may enhance the financial conditions of rural households while it could reduce the degree of forest dependency inspired with other income generating activities in due course.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.1
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• pp.30-39
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• 2014

HanPoong Leading (HPL)-04 were prepared with different oriental medicines (balk of Kalopanax pictus balk, Chaenomelis Fructus, Angelica gigas root, Zingiber officinale, Raphanus sativus Linne and Saururus chinensis Baill.) to investigate the protective effects of HPL-04 on cartilage degradation in knee osteoarthritis (OA). Rat articular chondrocytes incubated with rhIL-$1{\alpha}$ markedly increased matrix metalloproteinase (MMP)-2 and 9 activities, decreased cell viability and reduced chondrogenic gene expression. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, MMP-2 and 9 activities and real time RT-PCR indicated that HPL-04 counteracted these harmful effects in dose-dependent manner. In addition, for experimental OA in vivo, monosodium iodoacetate (MIA, 0.5 mg/50 ${\mu}L$) was injected into knee joints of rats and administered HPL-04 to rats for 4 consecutive weeks after MIA treatment. The experimental data showed that treatment with HPL-04 significantly prevented of MMP-2 and 9 activities in articular cartilage. Histopathological and micro-CT evaluations of the knee joints also revealed that HPL-04 effectively ameliorated MIA-induced degenerative OA. In conclusion, HPL-04 has potential applicability for the prevention and treatment of degenerative OA.

• Proceedings of the Korean Vacuum Society Conference
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• 2010.02a
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• pp.457-457
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• 2010

Flexible organic light emitting diodes (F-OLEDs) requires excellent moisture permeation barriers to minimize the degradation of the F-OLEDs device. Specifically, F-OLEDs device need a barrier layer that transmits less than $10^{-6}g/m^2/day$ of water and $10^{-5}g/m^2/day$ of oxygen. To increase the life time of F-OLEDs, therefore, it is indispensable to protect the organic materials from water and oxygen. Severe groups have reported on multi-layerd barriers consisting inorganic thin films deposited by plasma enhenced chemical deposition (PECVD) or sputtering. However, it is difficult to control the formation of granular-type morphology and microscopic pinholes in PECVD and sputtering. On the contrary, atomic layer deoposition (ALD) is free of pinhole, highly uniform, conformal films and show good step coverage. Thus, $Al_2O_3/TiO_2$ multi-layer was deposited onto the polyethersulfon (PES) substrate by electron cyclotron resonance atomic layer deposition (ECR-ALD), and the water vapor transmission rates (WVTR) were measured. WVTR of moisture permeation barriers is dependent upon density of films and initial state of polymer surface. A significant reduction of WVTR was achieved by increasing density of films and by applying low plasma induced interlayer on the PES substrate. In order to minimize damage of polymer surface, a 10 nm thick $TiO_2$ was deposited on PES prior to a $Al_2O_3$ ECR-ALD process. High quality barriers were developed from $Al_2O_3$ barriers on the $TiO_2$ interlayer. WVTR of $Al_2O_3$ by introducing $TiO_2$ interlayer was recorded in the range of $10^{-3}g/m^2.day$ at $38^{\circ}C$ and 100% relative humidity using a MOCON instrument. The WVTR was two orders of magnitude smaller than $Al_2O_3$ barriers directly grown on PES substrate without the $TiO_2$ interlayer. Thus, we can consider that the $Al_2O_3/TiO_2$ multi-layer passivation can be one of the most suitable F-OLEDs passivation films.

• Korean Journal of Food Science and Technology
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• v.50 no.1
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• pp.1-7
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• 2018

The MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay is commonly used for analyzing the cell viability. In this study, effects of various solvents, different lights, and zinc protoporphyrin (ZnPP) on the chemical behavior of MTT formazan were investigated. The color response of MTT formazan in NaOH was highly pronounced; the absorbance of MTT formazan in 0.1 N NaOH at 550 nm was >2-fold higher than that in water, dimethyl sulfoxide (DMSO), methanol, and ethanol. MTT formazan in DMSO and NaOH (>0.1 N) was relatively stable under fluorescent and UV light at 365 nm; its rapid degradation was induced under UV light at 254 nm in all solvents. ZnPP degraded MTT formazan under light in a time- and concentration-dependent manner; MTT formazan in 0.1 N NaOH was the most sensitive to ZnPP, followed by DMSO. These results suggest that NaOH and DMSO might be suitable media for MTT formazan for monitoring photosensitizing properties.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.1
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• pp.86-92
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• 2019

Di-ethylhexyl phthalate (DEHP) is an environmental contaminant that is used as a plasticizer. Endometriosis is a complex disease with an unknown etiology that is believed to be associated with exposure to DEHP. The present study examined the potential toxicity of DEHP by exposing endometrial adenocarcinoma cells (Ishikawa cells) to DEHP. In the experiments, the cells were treated with stepwise DEHP concentrations (0, 0.01, 0.1, 1, and $5{\mu}M$) for different exposure times (24, 48, and 72 h). When the relationship between the resulting survival rate of the cells and initial inflammation was examined, the cell viability, expression of $TNF-{\alpha}$, an inflammatory mediator, and the expression of MMP-9, an ECM degradation protein, were increased remarkably when the cells were exposed to $5{\mu}M$ DEHP for 48 and 72 hours. These results suggest that the exposure of Ishikawa cells to certain concentrations of DEHP, estrogen mimics, may cause time-dependent toxicity that affects the cell viability and inflammation, implying a potential role in the etiology of endometriosis. Further research on the effects of endocrine disruptors on the pathogenesis of endometriosis may reveal strategies to prevent this disease.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.47 no.4
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• pp.361-368
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• 2021

The objective of this study was to establish technology for removing bacteria with human- and eco-friendly material. Staphylococcus aureus as an important component for balanced equilibrium among microbiomes, was cultured under various concentrations of phosphate. Experimental observation relating to physical properties was performed in an addition of phosphate buffer. Statistically minimum value of size and hardness using atomic force microscope was observed on the matured biofilm at 5 mM concentration of phosphate. As a result of absorbance for the biofilm tagged with dye, concentration of biofilm was reduced with phophate, too. To identify whether this reduction by phosphate at the 5 mM is caused by counter ion or not, sodium chloride was treated to the biofilm under the same condition. To elucidate components of the biofilm counting analysis of the biofilm using time-of-flight secondary ion mass spectrometry was employed. The secondary ions from the biofilm revealed that alteration of physical properties is consistent to the change of extracellular polymeric substrate (EPS) for the biofilm. Viscoelastic characterization of the biofilm using a controlled shear stress rheometer, where internal change of physical properties could be detected, exhibited a static viscosity and a reduction of elastic modulus at the 5 mM concentration of phosphate. Accordingly, bacteria at the 5 mM concentration of phosphate are attributed to removing the EPS through a reduction of elastic modulus for bacteria. We suggest that the reduction of concentration of biofilm induces dispersion which assists to easily spread its dormitory. In conclusion, it is elucidated that an addition of phosphate causes removal of EPS, and that causes a function of antibiotic.