• Tuberculosis and Respiratory Diseases
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• v.45 no.1
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• pp.36-44
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• 1998

Background: IFN-$\gamma$ is known to activate mononuclear phagocytes and to mediate host defense mechanism against some intracellular microorganisms, but little is known about anti-mycobacterial activity and mechanism of IFN-$\gamma$ in human. In this study, we investigated the role of IFN-$\gamma$ in the pathogenesis of tuberculosis by observing the effect of IFN-$\gamma$ on the phagocytosis of M.tuberculosis(MTB) and on the production of TNF-$\alpha$ by human pulmonary alveolar macrophage. Method: Pulmonary alveolar macrophage(PAM) were prepared with adhesion purification method from bronchoalveolar lavage fluid obtained from 8 persorn without active lung lesion and cultured($1{\times}10^6cells/ml$) with MTB($3{\times}10^7$ bacteria/ml) with or without IFN-$\gamma$(300U/ml), LPS(0.5ug/ml) and autologous serum(10%). After 2 hours, the percentage of PAM-phagocytosed MTB was counted after AFB staining(modified Kynion method). TNF-$\alpha$ production by PAM stimulated by IFN-$\gamma$(300U/ml), MTB($1{\times}10^6bacteria/ml$) and LPS(0.5ug/ml) for 24hours was measured in culture supernatant using ELISA method. The degree of phagocytosis of MTB by PAM stimulated with IFN-$\gamma$(300U/ml) and LPS(0.5ug/ml) for 24hours was also investigated. Results: IFN-$\gamma$ did not influence the phagocytosis of MTB by PAM(percentage of PAM-phagocytosed MTB: control: $22.1{\pm}4.9$, IFN-$\gamma$: $20.3{\pm}5.3$) and did not increase TNF-$\alpha$ production by PAM (control: $21{\pm}38pg/ml$, IFN-$\gamma$: $87{\pm}106pg/ml$), and the degree of phagocytosis of MTB by PAM pre-stimulated with IFN-$\gamma$ for 24 hours, was not increased (control: $24.5{\pm}9.5$, IFN-$\gamma$: $23.4{\pm}10.1$). Conclusion: IFN-$\gamma$ does not influence on the phagocytosis of MTB and TNF-$\alpha$ production by PAM.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.8
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• pp.1086-1091
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• 2011

Anti-oxidative activity and tyrosinase inhibitory activity of various ethanol extracts of Polygoni multiflori radix (PMR) and Cynanchi wilfordii radix (CWR) were compared to identify an anti-oxidant and whitening agent source from nature. We conducted an investigation into the anti-oxidant activities of PMR and CWR ethanol extracts by measuring total polyphenol content, total flavonoid content, and ABTS radical capacity. The total polyphenol contents of PMR and CWR were 17.31${\pm}$0.54 mg GA/eq g, and 2.75${\pm}$0.22 mg GA/eq g, respectively. The total flavonoid contents of PMR and CWR were 6.38${\pm}$0.39 mg naringine/eq g, and 1.34${\pm}$0.09 mg naringine/eq g, respectively. The 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) radical decolorization of PMR and CWR were 96.89${\pm}$0.21% at 1 mg/mL and 93.49${\pm}$0.76% at 50 mg/mL. Melanoma cells were cultured with the PMR and CWR ethanol extracts for 48 hr, and total melanin content as a final product and the activity of tyrosinase, a key enzyme, in melanogenesis, were estimated. The PMR and CWR ethanol extracts increased melanin content and tyrosinase activity in a dose-dependent manner. These results suggest that PMR and CWR ethanol extracts could be useful as a skin whitening agent.

• Korean Journal of Biological Psychiatry
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• v.14 no.2
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• pp.115-121
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• 2007

Objetives : Identification of target genes for ethanol in neurons is important for understanding its molecular and cellular mechanism of action and the neuropathological changes seen in alcoholics. The purpose of this study is to identify of altered gene expression after acute treatmet of ethanol in rat gliom cells. Methods : We used high density cDNA microarray chip to measure the expression patterns of multiple genes in cultured rat glioma cells. DNA microarrays allow for the simultaneous measurement of the expression of several hundreds of genes. Results : After comparing hybridized signals between control and ethanol treated groups, we found that treatment with ethanol increased the expression of 15 genes and decreased the expression of 12 genes. Upregulated genes included Orthodenticle(Drosophila) homolog 1, procollagen type II, adenosine A2a receptor, GATA bindning protein 2. Downregulated genes included diacylglycerol kinase beta, PRKC, Protein phosphatase 1, clathrin-associated protein 17, nucleoporin p58, proteasome. Conclusion : The gene changes noted were those related to the regulation of transcription, signal transduction, second messenger systems. modulation of ischemic brain injury, and neurodengeneration. Although some of the genes were previously known to be ethanol responsive, we have for the most part identified novel genes involved in the brain response to ethanol.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.195-200
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• 2015

The objective of this study was to evaluate the toxicities of permeable cryoprotectants and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Toxicities of permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG), Glycerol, and 1,2-PROH were investigated using a murine embryo model. Female $F-{_1}$ mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to among DMSO, EG, Glycerol, and 1,2-PROH. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO and Glycerol at the 2-cell stage was significantly lower than that were treated with EG ($81.1{\pm}15.1$), 1,2-PROH ($88.0{\pm}21.1$) or the control ($99.9{\pm}21.3$) (p<0.001). On comparison of four cryoprotectant treated groups, the DMSO and Glycerol treated group showed a decreased cell count compared with the EG and 1,2-PROH treated group (p<0.05). Both DMSO ($14.7{\pm}1.3$), EG ($12.1{\pm}1.1$), Glycerol ($15.2{\pm}1.8$), and 1,2-PROH ($11.5{\pm}1.3$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.5{\pm}0.7$, p<0.0001). In addition, the DMSO or Glycerol treated group showed more apoptotic cells than the EG or 1,2-PROH treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to among DMSO, EG, Glycerol, and 1,2-PROH at room temperature. When comparing four permeable cryoprotective agents, EG and 1,2-PROH appeared to be less toxic than DMSO and Glycerol at least in a murine embryo model.

• The Korean Journal of Nuclear Medicine
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• v.35 no.3
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• pp.168-184
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• 2001

Purpose: KR-30035 (KR), a new MDR reversing agent, has been found to produce a similar degree of increased Tc-99m MIBI uptake in cultured tumor cells over-expressing mdr1 mRNA compared to verapamil (VP), with less cardiovascular effects. We assessed the MDR-reversing ability of KR in vivo, and effects of various doses of KR on MIBI uptake un nude mice hearing P-glycoprotein (P-gp) positive (+) and P-gp negative (-) human tumor xenografts. Methods: P-gp (+) HCT15/CL02 colorectal and P-gp (-) A549 non-small cell cancer cells were inoculated in each flank of 120 nude mice (20 mice ${\times}$ 6 groups). Group 1 (Gr1) mice received 10mg/kg KR i.p. 3 times $({\times}3)$; Gr2, 10mg/kg VP i.p. ${\times}3$; Gr3, 10mg/kg KR i.p. ${\times}2$ + 25mg/kg KR i.p. ${\times}1$; Gr4, 10mg/kg KR i.p. ${\times}2$ + 50mg/kg i.p. ${\times}1$; Gr5, 10mg/kg KR i.p. ${\times}2$ + 25mg/kg KR i.v. ${\times}1$, GrC, controls. The mice were then injected with Tc-99m MIBI and sacrificed after 10 min, 30 min, 90 min and 240 min. Tumor uptake of MIBI (TU) in each group was compared. Results: TU in P-gp (+) and (-) tumors were both higher in Gr1 than Gr2. Washout rate between the 10 min and 4 hours was lower in Gr5 of P-gp (+) cell(0.93) than the control. Percentage increases in TU were higher in P-gp (+) than P-gp (-) tumors with all KR doses. Pgp (+) TU were highest at 10 mon (173% of GrC) and persisted up to 240 min (144%) in Gr3. Larger doses of KR resulted in a lesser degree of increase in P-gp (+) TU at 10 min (130% in Gr4 and 117% un Gr5) and 30 min (178%, 129%), but TU increased by time up to 240 min (177%, 196%). Heart and lung uptakes were markedly increased in Gr4 and Gr5 at 10 and 30 min, likely due to cardiovascular effects. No mice died. Conclusion: These data further suggest that KR that has significantly lower cardiovascular toxicity than verapamil can be used as an active inhibitor of MDR. Even a relatively low dose of KR significantly increased Tc-99m MIBI uptake in P-gp (+) tumors in vivo.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.6
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• pp.527-535
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• 2020

The purpose of this study was to establish a simple vitrification protocols to preserve animal cell lines derived from tissues of livestock that could be recultured. Bovine oviduct epithelial cells (BOEC) were used for the vitrification process using a 0.25 ml straw to increase cryopreservation efficiency. BOEC was cultured from the oviduct of 3.5-day estrus state, and the commercially available polyampholyte StemCell Keep^TM was used as a cryoprotective agent. Using different concentrations, the viability rates of BOEC in 5, 10, 25, 50, 75, and 100% in freezing media were investigated. Survivability was determined using a differential staining technique using a trypan blue test and a CYTO-13/PI staining protocol. The viability rates of BOEC in the trypan blue test were 5.6±11.8, 12.5±7.2, 53.0±2.7, 85.1±6.9, 79.8±0.6, and 60.7±6.7% with a respective concentration of StemCell Keep^TM. The viability rates in CYTO-13/PI staining were 4.6±2.5, 30.8±12.1, 58.4±2.5, 85.5±1.2, 79.8±0.6, and 71.2±1.2%, respectively. These results indicate that BOEC could be preserved with StemCell Keep^TM without toxicity in a 0.25-ml straw. The optimal concentration of vitrification solution with StemCell Keep^TM was determined to be 50% and can be considered as a proper preservation method for cryobanking.

• Journal of the Korean Chemical Society
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• v.51 no.3
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• pp.251-257
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• 2007

The Tumor necrosis factor-α, (TNF-α), is involved in the pathogenesis of multiple sclerosis and contributes to the degeneration of oligodendrocytes as well as neurons. Nicotine has been found to have immunosuppressive and inflammation-suppressing effects. Astrocytes, the major glial cells in the CNS, are capable of producing TNF-α at both the mRNA and protein levels in response to interleukin-1 (IL-1) or TNF-α. Nicotine has been shown to influence glial cell functions. To order to explore the role of astrocytes in the production of TNF-α, astrocytes were pretreated with nicotine and are stimulated with IL-1β to determine their effects on TNF-α production. The results are as follows. Cytotoxic effects of nicotine on human fetal astrocytes were noted above 10 μg/ml of nicotine. The effect of IL-1β on TNF-α mRNA expression in primary cultured human fetal astrocytes was maximal at 2 h after IL- 1β(100 pg/ml) treatment. Human fetal astrocytes were pretreated with 0.1, 1, and 10 μg/ml of nicotine and then stimulated with IL-1β (100 pg/ml) for 2 h. The inhibitory effect of nicotine on expressions of TNF-α mRNA in human fetal astrocytes with pretreated 0.1 μg/ml of nicotine is first noted at 8 hr, and the inhibitory effect is maximal at 12 h. The inhibitory effect at 1 μg/ml of nicotine is inhibited maximal at 24 h. Nicotine at 0.1, 1 and 10 μg/ml concentrations significantly inhibits IL-1β-induced NF-κB activation. Collectively, this study indicates that nicotine might inhibit the expression of TNF-α in activated human fetal astrocytes.

• The Korean Journal of Ecology
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• v.17 no.3
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• pp.299-310
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• 1994

Intraspecific clonal interactions have important influences on a population structure of the cellular slime mold (CSM). This study was to investigate whether or not evolutionary change in a population could be induced by clonal competition, and to elucidate how various clones in a population evolve in a homogeneous environment of laboratory culture. The characteristic clones of Polysphondylium pallidum which had different resource consumption rates (RCR) and mating types I and II were selected for study. Investigation was conducted for 4 experimental time interval $(T_0-T_4)$; one experimental time interval took almost 10-14 days from inoculation to havest of fruiting bodies. Two sets of 50 clones were cultured from 50 clones at To, and RCR variations of the population were compared between $(T_0\;and\;T_4)$ for each set of clones. Each clone of the CSM had a diverse resource consumption rate, or growth rate, in a homogeneous and limited Cerophyl agar plate despite the passage of 48-56 generations from the beginning of the experiment. Diverse clones with different growth rate could coexist in one site of the homogeneous agar plate as well as heterogeneous soil microenvironment. When there was high clonal diversity of RCR, a clone in a population had high chances to encounter other clones with resultant increased clonal competition. In one set, 26 of 37 clones of mating type I were changed to mating type Il for the 4 experimental time intervals, which indicated that the rate of competitive exclusion among clones during total experiment from $(T_0\;to\;T_4)$ was 0.703. In another set, 31 of 37 clones of mating type I were changed to mating type II , having the rate of competitive exclusion 0.838. The frequency of each of mat~ng types changed by 0.93-1.29% in each successive generation. The competitive exclusion among clones occurred by 1.26-1.75% when approximately $2.6{\times}10^8$ bacterial cells were provided as food and thereafter one generation of myxamoebae of CSM elapsed at room temperature. This finding implicated that in the vegetative state of P, pallidurn there was 1.26-1.75% probabil~ty of evolutionary change per generation changing from one clone to another clone.

• Journal of the korean academy of Pediatric Dentistry
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• v.27 no.1
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• pp.15-23
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• 2000

Streptococcus salivarius is a normal inhabitant in the human oral cavity. Streptococcus salivarius 119 in this study was isolated from the oral cavity of child and identified, and its action mechanism on the formation of denal plaque by Streptococcus matans was studied. 1. The optical density of absorbance at 550 nm was 0.327 in the culture of Streptococcus mutans in disposable cuvette, whereas being 0.119 in the combined culture of Streptococcus mutans and Streptococcus salivarius 119. 2. The mean weight of produced artificial plaque on the wires in the beaker was 84.7mg in culture of Streptococcus mutans only, whereas being reduced to 12.3mg in the combined culture of Streptococcus mutans and Streptococcus salivarius 119. 3. When Streptococcus mutans was cultured in the media containing culture supernatant of Streptococcus salivarius 119 in BHI broth, the mean weight of produced artificial plaque was 100.5mg on the wires, whereas being reduced to 20.4mg in the media containing culture supernatant of Streptococcus salivarius 119 in BHIS broth. 4. The viable cells of Streptococcus oralis and Streptococcus salivarius 119 were $4.8\times10^7\;and\;7.5\times10^8$ per ml respectively after each culture, wheras being $4.2\times10^7\;and\;5.8\times10^7$ per ml in the combined culture of Streptococcus oralis and Streptococcus salivarius 119. 5. The polymer produced by Streptococcus salivarius 119 was glucan on the thin layer chromatography. 6. The glucan produced by Streptococcus salivarius 119 was water-soluble glucan containing $1\rightarrow6$ linkages as the main linkage on the thin layer chromatography. These results suggested that isolated Streptococcus salivarius 119 inhibited the formation of artificial plaque by the production of water-soluble glucan.

• The Journal of Korean Academy of Prosthodontics
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• v.50 no.1
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• pp.44-52
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• 2012

Purpose: In this paper we tried to evaluate the most appropriate surface for rhBMP-2 coating among 4 rough titanium surfaces. Materials and methods: We used machined surface as a control group and anodized, RBM and SLA surfaces as test groups. We coated rhBMP-2 on the 4 surfaces and with uncoated surfaces for each case, we cultured human mesenchymal stem cells on all 8 surfaces. 24 hours after we measured the stem cell' attachment with SEM, and on 3rd, 7th, and 14th days, we checked the cell proliferation and differentiation by using MTT and ALP activity assay. And on the 7th day after the culture, we performed RT-PCR assay to determine whether the expression levels of Type I collagen, osteocalcin, osteopontin were changed. Results: We observed with SEM that 4 rhBMP-2 coated surfaces exhibited wider and tighter cell attachment and more cell process spreading than uncoated surfaces. The anodized rhBMP-2 surface caused robustest effects. In MTT assay we could not find any meaningful difference. In ALP assay there was a significant increase (P<.05) in the ALP activity of anodized rhBMP-2 coated surface compared with that of the control (3rd and 14th days) and with that of the RBM rhBMP-2 coated surface (14th day). In RT-PCR assay there was increased expressions in the anodized rhBMP-2 coated surface for osteocalcin, and osteopontin. Conclusion: We found that the anodized rhBMP-2 coated surface were most prominent stem cell attachment and differentiation in compared to control and Machined rhBMP-2 coated, RBM rhBMP-2 coated surface.