• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2002.10a
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• pp.18-25
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• 2002

The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.17 no.3
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• pp.262-269
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• 2016

The aim of this study was to identity the death anxiety of elderly people living alone, and the factors that affect their death anxiety. The subjects did not have an impaired cognitive function, and were from one Chungcheongnamdo 2 district in the elderly University. The subjects were 187 elderly people over the age of 65 living alone in the district. The character, communication, and data was collected from February, 2014 to 2 May, 2015 and analyzed using the SPSS 18 program. The death anxiety whole point of elderly people living alone was 2.94 (${\pm}0.32$); it was 3.06 (${\pm}0.32$) points according to the sub-region 'death process anxiety', 2.88 (${\pm}0.51$) points according to 'after-death anxiety', and 2.75 (${\pm}0.43$) points according to 'presence loss anxiety'. The factors affecting the death anxiety were economic status, depression, and spiritual wellbeing. Economic status (${\beta}=-.36$, p= .000) had the largest effect with an overall explanatory power of 20.3%. Therefore, for the death anxiety of elderly people living alone, life needs to be strengthened through social security systems with intervention programs to improve the quality of depression and spiritual wellbeing.

• The Korean Journal of Pharmacology
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• v.28 no.1
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• pp.91-102
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• 1992

In $Ca^{++}$ containing media, arachidonic acid markedly stimulated superoxide and $H_2O_2$ generation and activated NADPH oxidase. In $Ca^{++}$ free media, stimulatory action of arachidonic acid on NADPH oxidase was not detected. Arachidonic acid-stimulated respiratory burst was inhibited by EGTA, TMB-8, verapamil, diltiazem, nifedipine, dibucaine, lidocaine, CCCP, 2,4-dinitrophenol, sodium arsenate, chlorpromazine, theophylline, $HgCl_2$, PCMB and PCMBSA but not affected by tetrodotoxin, tetraethylammonium chloride and procaine. EGTA almost completely inhibited release of ${\beta}-glucuronidase$ by arachidonic acid and verapamil, CCCP and theophylline slightly inhibited it, whereas dibucaine did not show any significant effect. Arachidonic acid induced $Ca^{++}$ release from intact neutrophils and it was decreased by TMB-8. Arachidonic acid-induced elevation of intracellular free $Ca^{++}$ level was inhibited by EGTA and CCCP and slightly inhibited by TMB-8. Amount of intracellular free $Ca^{++}$ increased by either arachidonic acid plus verapamil or arachidonic acid plus dibucaine was greater than that by arachidonic acid alone. These results suggest that various changes of biochemical events may be implicated in the functional expression in neutrophils activated by arachidonic acid. Arachidonic acid appears to elevate cytosolic free $Ca^{++}$ level by stimulating $Ca^{++}$ release from intracellular $Ca^{++}$ storage sites. During activation of neutrophils, $Ca^{++}$ influx and efflux may be accomplished, simultaneously.

• The Korean Journal of Pharmacology
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• v.24 no.1
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• pp.83-92
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• 1988

Although the release of lysosomal enzymes from activated PMN leukocyte can be regulated by intracellular cyclic nucleotide levels, other responses of PMN leukocyte according to the binding of neurotransmitters to either ${\beta}$-adrenergic or muscarinic receptors are still not clarified. In addition, the function of PMN leukocyte mediated by ${\alpha}$-adrenergic receptors is uncertain. Atropine, phentolamine and propranolol inhibited calcium uptake, superoxide generation, NADPH oxidase activity and phagocytic activity in activated PMN leukocyte, whereas carbachol and isoproterenol slightly further stimulated the responses of activated cells. Either carbachol or isoproterenol stimulated superoxide generation was inhibited by their antagonists, atropine and propranolol, respectively. The response of activated PMN leukocyte was inhibited by chlorpromazine, verapamil and dantrolene but slightly stimulated by lithium. On the other hand, chlorpromazine and dibucaine did not affect NADPH oxidase activity. Atropine, phentolamine and propranolol suppressed the calcium dependent phagocytic activity. Thus, the results suggest that atropine, phentolamine and propranolol may inhibit superoxide generation in activated PMN leukocyte by the inhibition of calcium influx and by their direct action on the NADPH oxidase system which is associated with autonomic receptors.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.403-412
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• 2014

To develop an effective skin whitening agent for cosmetics, we isolated cucurbitacin B from Cucumis sativus L. which has been used as traditional skin lighting regimen by the bioactivity-guided fractionation, and investigated the inhibitory effects of cucurbitacin B on melanogenesis. At a non-cytotoxic concentration, cucurbitacin B reduced melanin contents of B16F1 melanoma cells in a dose-dependent manner. Cucurbitacin B did not directly inhibit mushroom tyrosinase activity, but it inhibited intracellular tyrosinase activity in a dose-dependent manner. Its inhibitory mechanism on melanin biosynthesis was further assessed, and we found that cucurbitacin B significantly decreased the protein level of tyrosinase, a major melanogenic enzymes and MITF, a master transcriptional factor of melanogenesis. In addition, cucurbitacin B increased the expression of WW domain-containing oxidoreductase (WWOX) which is known to function as tumor repressor and inhibits $Wnt/{\beta}$-catenin pathway. Collectively, these results suggest that cucuritacin B from C. sativus could be used as an active ingredient for skin whitening.

• Journal of the Institute of Electronics and Information Engineers
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• v.50 no.3
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• pp.146-152
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• 2013

In this paper, we have proposed and experimentally demonstrated an intensity-based self-referencing fiber optic sensor. The proposed fiber optic sensor consists of a broadband light source (BLS), fiber Bragg grating (FBG), tunable Fabry-Perot (F-P) filter, and LabVIEW program. We define the measurement parameter (X) and the calibration parameter (${\beta}$) to determine the transfer function(H) of the self-referencing fiber optic sensor, and the validity of the theoretical analysis is confirmed by experiments. The self-referencing characteristic for the proposed system has been validated by showing that the measurement parameter (X) is invariant for BLS optical power attenuations of 0 dB, 3 dB, and 6 dB. Also, the measured result is irrelevant to the FBGs with different characteristics. This means that the proposed fiber optic sensor offers the flexibility for determining the FBGs needed for implementation. Experimental results for the proposed fiber optic sensor are in good agreement with a theoretical analysis for BLS optical power attenuations and for three FBG pairs with different characteristics. So, the proposed fiber optic sensor has several benefits, including the self-referencing characteristic and the flexibility to determine the FBGs.

• Biomolecules & Therapeutics
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• v.9 no.3
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• pp.167-175
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• 2001

Recent studies have shown that cytokines are capable of modulating cardiovascular function and that some drugs used in the treatment of heart failure variably modulate the production of cytokines. Hige- namine, a positive inotropic isoquinoline alkaloid, has been used traditionally as cardiac stimulant, and reported to reduce nitric oxide (NO) and inducible nitric oxide synthase (iNOS) expression in LPS- and/or cytokine-activated cells in vitro and in vivo. Therefore, we investigated whether higenamine modulates the production of proinflammatory cytokines in myocardial infarction. In addition, effects of higenamine on antioxidant action and antioxidant enzyme expression (MnSOD) were studied. Myocardial infarction (MI) was confirmed by measuring left ventricular (LV) pressure after occlusion of the left anterior descending coronary artery (LAD) for 5 weeks in rats. Treatment of higenamine (10 mg/kg/day) reduced infarct size about 35 %, which accompanied by reduction of production TNF-$\alpha$, IL-6, but not IFN-${\gamma}$ and IL-1$\beta$ in the myocardium. The expression of TNF-$\alpha$ mRNA in infracted myocardium was significantly reduced by higenamine. Although iNOS mRNA was not detected, nitrotyrosine staining was significantly increased in myocardium of Ml compared to higenamine-treated one, Indicating that peroxynitrite-induced damage is evident in MI. Cytochrome c oxidation by peroxynitrite was concentration-dependently reduced by higenamine, an effect which was almost compatible to glutathion. Higenamine treatment did not affect the expression of MnSOD mRNA in myocardial tissues in MI. Taken together, higenamine may be beneficial in oxidative stress conditions such as ischemic-reperfusion injury and MI due to antioxidant action as well as modulation of cytokines.

• Journal of the Korean Chemical Society
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• v.56 no.5
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• pp.556-562
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• 2012

The critical micelle concentration (CMC) and the counter-ion binding constant (B) for the mixed micellizations of TTAB (tetradecyltrimethylammonium bromide) with other surfactants (DTAB, CTAB, Tween-20, Tween-40, and Tween-80) in aqueous solution of 4-chlorobenzoic acid (0.5 mM) at $25^{\circ}C$ were determined as a function of ${\alpha}_1$ (the overall mole fraction of TTAB) by using the spectrophotometric method and the conductivity method. Various thermodynamic parameters ($X_i$, ${\gamma}_i$, $C_i$, $a_i^M$, ${\beta}$, and ${\Delta}H_{mix}$) were calculated for each mixed surfactant system and compared with the other mixed surfactant systems by means of the equations derived from the nonideal mixed micellar model. The results show that TTAB/DTAB mixed system has a great positive deviation from the ideal mixed micellar model and the other mixed systems have great negative deviations from the ideal mixed model.

• Journal of Korea Port Economic Association
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• v.31 no.1
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• pp.1-14
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• 2015

The standard GARCH model imposing symmetry on the conditional variance, tends to fail in capturing some important features of the data. This paper, hence, introduces the models capturing asymmetric effect. They are the EGARCH model and the GJR model. We provide the systematic comparison of volatility models focusing on the asymmetric effect of news on volatility. Specifically, three diagnostic tests are provided: the sign bias test, the negative size bias test, and the positive size bias test. This paper shows that there is significant evidence of GARCH-type process in the data, as shown by the test for the Ljung-Box Q statistic on the squared residual data. The estimated unconditional density function for squared residual is clearly skewed to the left and markedly leptokurtic when compared with the standard normal distribution. The observation of volatility clustering is also clearly reinforced by the plot of the squared value of residuals of export volume and values. The unconditional variance of both export volumes and export value indicates that large shocks of either sign tend to be followed by large shocks, and small shocks of either sign tend to follow small shocks. The estimated export volume news impact curve for the GARCH also suggests that $h_t$ is overestimated for large negative and positive shocks. The conditional variance equation of the GARCH model for export volumes contains two parameters ${\alpha}$ and ${\beta}$ that are insignificant, indicating that the GARCH model is a poor characterization of the conditional variance of export volumes. The conditional variance equation of the EGARCH model for export value, however, shows a positive sign of parameter ${\delta}$, which is contrary to our expectation, while the GJR model exhibits that parameters ${\alpha}$ and ${\beta}$ are insignificant, and ${\delta}$ is marginally significant. That indicates that the asymmetric volatility models are poor characterization of the conditional variance of export value. It is concluded that the asymmetric EGARCH and GJR model are appropriate in explaining the volatility of export volume, while the symmetric standard GARCH model is good for capturing the volatility.

• Development and Reproduction
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• v.2 no.1
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• pp.45-52
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• 1998

An enhncer detector line(EDL) having P[1ArB] insertion in X chromosome with expression of reporter gene (lacZ) in the polar cells and border cell of egg chamber was established and used to monitor the differentiation and migration of border cells during the oogenesis of Drosophila. differentiation of border cell from the anterior polar follicle cells was evident in stage-9 egg chamber of EDL149 which was characterized by migration of columnar follicle cells toward posterior of egg chamber surrounding the oocyte. Migration of border cells was observed in the stage-9 and -10 egg chambers. \beta -galactosidase activities were rapidly increased during the first 4 days after eclosion, and it coincided with the timing of border cell differentiation in the ovary during adult life. Homozygote of EDL149 showed some retardation of border cell migration , resulting absence of migration of some border cells in the anterior part of egg chamber or delayed migration of some border cells in the stage-10 egg chamber. These results suggest that the P[1ArB] of EDL149 is inserted at the locus of the structural gene required for the border cell migration. In addition to the expression in egg chambers, lacZ expression was also detected in the meiotic germ cells of testis and antenna, suggesting the possible requirement of the trapped gene function in these organ. this EDL and enhancer trapped gene might be useful for the study of developmentally regulated cell migration.