• Journal of Life Science
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• v.25 no.6
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• pp.631-637
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• 2015

Streptavidin, a protein produced by Streptomyces avidinii, strongly binds up to four molecules of vitamin H, d-biotin exhibiting the dissociation constant of about 10⁻¹⁵ M. This strong binding affinity has been applied for detection and characterization of numerous biological molecules suggesting expression and purification of functional streptavidin should be very useful for the application of this streptavidin-biotin interaction. To express a soluble streptavidin in Escherichia coli, We synthesized streptavidin genes and cloned into pET-22b plasmid, which uses T7 RNA polymerase/T7 promoter expression systems containing pelB leader for secretion into periplasmic space and six polyhistidine tags at C-terminus for purification of expressed proteins. Although streptavidin is toxic to Escherichia coli due to strong biotin binding property, streptavidin was expressed very sufficiently in a range of 10-20 mg/ml. In SDS-PAGE, the size of purified protein was shown as 17 kDa in denatured condition (boiling) and 68 kDa in native condition (without boiling) suggesting tetramerization of monomeric subunit by non-covalent association. Further analysis by size-exclusion chromatography supported streptavidin’s tetrameric structure as well. In addition, soluble streptavidin detected biotinylated proteins in westernblot indicating its functional activity to biotin. Taken these results together, it concluded that our simple expression system was able to show high yield, homotetrameric formation and biotin binding activity analogous to natural streptavidin.

• Korean Journal of Agricultural and Forest Meteorology
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• v.18 no.1
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• pp.16-29
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• 2016

The total area of paddy field was estimated to be 55 % of the cultivated lands in South Korea, which is approximately 1 million hectares. Organisms inhabiting paddy fields if they are sensitive to environmental changes can be environmental indicator of paddy fields. Biological indicators such as phenology and distributional range are evaluated as intuitive and quantitative method to analyze the impact of climate change. This study aims to estimate flight time change of Hydrophilidae species' based on the RCP 8.5 climate change scenario. Unmanned monitoring systems were installed in Haenam, Buan, Dangjin and Cheorwon relative to the latitudinal gradient. In the three regions excepting Cheorwon, it was able to measure the abundance of flying Hydrochara affinis and Sternolophus rufipes. Degree-day for the flight time was determined based either on field measurement values and estimates of 2020s, 2050s and 2080s from KMA climate change scenario data. As a result, it is found that date of both species of initial flight becomes 15 days earlier, that of peak flight becomes 22 days earlier and that of final flight does 27 days earlier in 2080s compared to 2020s. The climate change impact on flight time is greater in coastal area, rural area and valley than inland area, urban area and plan. H. affinis and S. rufipes can be used as climate change indicator species.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.5
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• pp.389-399
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• 2015

Zinc has been considered as a vital constituent of proteins, including enzymes. Mobile reactive zinc ($Zn^{2+}$) is the key form of zinc involved in signal transductions, which are mainly driven by its binding to proteins or the release of zinc from proteins, possibly via a redox switch. There has been growing evidence of zinc's critical role in cell signaling, due to its flexible coordination geometry and rapid shifts in protein conformation to perform biological reactions. The importance and complexity of $Zn^{2+}$ activity has been presumed to parallel the degree of calcium's participation in cellular processes. Whole body and cellular $Zn^{2+}$ levels are largely regulated by metallothioneins (MTs), $Zn^{2+}$ importers (ZIPs), and $Zn^{2+}$ transporters (ZnTs). Numerous proteins involved in signaling pathways, mitochondrial metabolism, and ion channels that play a pivotal role in controlling cardiac contractility are common targets of $Zn^{2+}$. However, these regulatory actions of $Zn^{2+}$ are not limited to the function of the heart, but also extend to numerous other organ systems, such as the central nervous system, immune system, cardiovascular tissue, and secretory glands, such as the pancreas, prostate, and mammary glands. In this review, the regulation of cellular $Zn^{2+}$ levels, $Zn^{2+}$-mediated signal transduction, impacts of $Zn^{2+}$ on ion channels and mitochondrial metabolism, and finally, the implications of $Zn^{2+}$ in health and disease development were outlined to help widen the current understanding of the versatile and complex roles of $Zn^{2+}$.

• Journal of the Korean Applied Science and Technology
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• v.27 no.4
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• pp.559-567
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• 2010

In this study, the antioxidative effect, cellular protective effect and inhibitory effect on tyrosinase of Cosmos bipinnatus extracts were investigated. The ethyl acetate fraction of Cosmos bipinnatus flower extract ($11.48\;{\mu}g$/mL) showed more excellent free radical (1,1-diphenyl-2-picrylhydrazyl radical, DPPH) scavenging activity (FSC50) than those of leaf and stem extracts ($17.45\;{\mu}g$/mL). Reactive oxygen species (ROS) scavenging activity (OSC50) of Cosmos bipinnatus extracts on ROS generated in $Fe^{3+}$-EDTA/H2O2 system were investigated using the luminol-dependent chemiluminescence assay. The ethyl acetate fraction of Cosmos bipinnatus flower extract ($0.56\;{\mu}g$/mL) showed 3 times more excellent ROS scavenging activity than L-ascorbic acid ($1.50\;{\mu}g$/mL). The protective effects of the ethyl acetate fractions of extracts from different parts of Cosmos bipinnatus on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethyl acetate fractions of leaf and stem extract and flower extracts suppressed photohemolysis in a concentration dependent manner ($10\sim50\;{\mu}g$/mL). The inhibitory effect of ethyl acetate fraction of Cosmos bipinnatus flower extract ($62.75\;{\mu}g$/mL) on tyrosinase was investigated to assess the whitening efficacy. The ethyl acetate fraction of Cosmos bipinnatus flower extract showed 3.5 times higher tyrosinase inhibitory effect than arbutin ($226.88\;{\mu}g$/mL) known as an effective whitening agent. These results indicate that fractions of Cosmos bipinnatus extracts can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O2$ and other ROS, and protect cellular membranes against ROS. Fractions of Cosmos bipinnatus extracts can be applicable to new functional cosmetics for antioxidant and whitening.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.4
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• pp.651-656
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• 2001

Ascorbic acid(AsA) is unevenly distributed throughout all body cells and fluids. Multiactivities of AsA in many biological systems and in various scientific fields were reported. In this study we aimed to clarify the inhibitory action of high concentration of AsA on the cell growth in 3T6 fibroblasts. The cells wee exposed to AsA at various concentration. It showed that 3T6 fibroblasts wee dead by the medium which contained AsA at the concentration higher than 0.5 mM. AsA caused hydrogen peroxide ($H_2O$$_2$) generation in a concentration dependent manner. These results suggested that the $H_2O$$_2$ was formed in the medium by AsA and acted as a cytotoxic gent. Moreover, it is supposed that hydroxyl radical (.OH) induced from $H_2O$$_2$also acetd as actively cytotoxic agent. This lethal effect of AsA causing the cell death was inhibited by the addition of catalase in the medium. Therefore, addition of AsA at the normal concentrations stimulate cell growth, but excess concentrations of AsA induce cell death.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.33 no.2
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• pp.61-67
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• 2007

In this study, the antioxidative effects of Equisetum arvense extracts were investigated. The free radical (1,1 diphenyl-2-picrylhydrazyl, DPPH) scavenging activity ($FSC_{50}$) of extract/fractions of Equisetum arvense was in the order: 50 % ethanol extract ($182.04{\mu}g/mL$) < ethylacetate fraction ($54.50{\mu}g/mL$) < deglycosylated flavonoid aglycone fraction ($14.13{\mu}g/mL$). Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of some Equisetum arvense extracts on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol- dependent chemiluminescence assay. The order of ROS scavenging activity was deglycosylated flavonoid aglycone fraction ($OSC_{50}$, $3.54{\mu}g/mL$) < 50 % ethanol extract ($0.80{\mu}g/mL$) < ethylacetate fraction ($0.006{\mu}g/mL$). Ethylacetate fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Equisetum arvense on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The ethanol extract (50%) suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated aglycone extract exhibited the most prominent celluar protective effect ($\tau_{50}$, 161.10 min at $10{\mu}g/mL$). These results indicate that extract/fractions of Equisetum arvense can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging $^1O_2$ and other ROS, and protect cellular membranes against ROS.

• KSBB Journal
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• v.24 no.4
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• pp.397-402
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• 2009

The Portulaca oleracea (P. oleracea) is a popular herbal medicine in East Asia that was known to possess detoxification, antifebrile and antifungal effects. In the present study, we examined the biological activities of ethanol extracts of P. oleracea under various conditions with NIH3T3, B16F10, and MCF-7 cell line model systems. Extracts of P. oleracea (0.5 mg/ml) showed inhibition of expression of tyrosinase, but does not suppress either of TYRP-1 or DCT expression on B16F10 cells. Extracts of P. oleracea (2 mg/ml) showed anti-inflammatory effects on TNF-$\alpha$-stimulated NIH3T3/$NF{\kappa}B$-Luc cells and increase of the synthesis of collagen on NIH3T3 (wild type) cells. These results suggest that extracts of P. oleracea could be used as a functional biomaterial in developing a skin whitening agent and having the anti-inflammatory, anti-wrinkle, and anti-aging activities.

• Advances in Traditional Medicine
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• v.3 no.3
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• pp.157-162
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• 2003

Various extracts of higher plants have been used in traditional medicine systems for centuries. While tropical and sub-tropical plants have received considerable attention from the researchers for evaluation of their bioactivity, temeperate plants have always been neglected somewhat. Similarly, seeds of the plants have not been considered seriously compared to other plant parts, e.g. leaves, stems, roots, flowers, etc. as a potential source for biologically active compounds. As part of our on-going evaluation of the extracts of the seeds of temperate plants, especially from Scotland, for biological activity, Achillea millefolium, Angelica sylvestris and Phleum pratense have been chosen for the present study. Both A. millefolium and A. sylvestris are well known for their traditional medicinal uses in Europe and also in the orient, but there is no report on any medicinal properties of P. pratense available to date. Extracts of the seeds of these plants have been assessed for their antioxidant and antibacterial potential and also for general toxicity. Both DCM and MeOH extracts of A. millefolium showed the most significant broad spectrum antibacterial activity among the three plants and inhibited the growth of almost all test strains of bacteria. The DCM extracts of all three species were active against methicillin resistant Staphylococcus aureus (MRSA) and Citrobacter freundii $(MIC=6.25{\times}10^{-1}\;mg/mL)$. While the MeOH extracts of A. millefolium and P. pratense were active against C. freundii, that of P. pratense was also active against MRSA. The MeOH extract of A. sylvestris did not show any antibacterial activity against any of the eight bacterial strains at test concentrations. The MeOH extract of P. pratense showed the most prominent antioxidant activity $(IC_{50}=145\;{\mu}g/ml)$ and there was no antioxidant activity observed with the DCM extract of A. millefolium. The DCM extract of P. pratense was the most toxic $(LC_{50}=20\;{\mu}g/ml)$ among the extracts.

• Molecules and Cells
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• v.41 no.8
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• pp.742-752
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• 2018

Parkinson's disease (PD) is a neurodegenerative disease characterized by progressive degeneration of dopaminergic (DAergic) neurons, particularly in the substantia nigra (SN). Although circadian dysfunction has been suggested as one of the pathophysiological risk factors for PD, the exact molecular link between the circadian clock and PD remains largely unclear. We have recently demonstrated that $REV-ERB{\alpha}$, a circadian nuclear receptor, serves as a key molecular link between the circadian and DAergic systems. It competitively cooperates with NURR1, another nuclear receptor required for the optimal development and function of DA neurons, to control DAergic gene transcription. Considering our previous findings, we hypothesize that $REV-ERB{\alpha}$ may have a role in the onset and/or progression of PD. In the present study, we therefore aimed to elucidate whether genetic abrogation of $REV-ERB{\alpha}$ affects PD-related phenotypes in a mouse model of PD produced by a unilateral injection of 6-hydroxydopamine (6-OHDA) into the dorsal striatum. $REV-ERB{\alpha}$ deficiency significantly exacerbated 6-OHDA-induced motor deficits as well as DAergic neuronal loss in the vertebral midbrain including the SN and the ventral tegmental area. The exacerbated DAergic degeneration likely involves neuroinflammation-mediated neurotoxicity. The $REV-erb{\alpha}$ knockout mice showed prolonged microglial activation in the SN along with the over-production of interleukin $1{\beta}$, a pro-inflammatory cytokine, in response to 6-OHDA. In conclusion, the present study demonstrates for the first time that genetic abrogation of $REV-ERB{\alpha}$ can increase vulnerability of DAergic neurons to neurotoxic insults, such as 6-OHDA, thereby implying that its normal function may be beneficial for maintaining DAergic neuron populations during PD progression.

• KSBB Journal
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• v.22 no.6
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• pp.409-413
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• 2007

Polydiacetylene(PDA)-based sensors possess a number of properties that can be successfully applied for label-free detection system. PDA is one of the most attractive color-generating materials, with growing applications as sensors. Here we introduce various PDA-based devices, used as biosensor, chemosensor, thermosensor, and optoelectronics sensor. In general, PDA liposomes and films are closely packed and properly designed for polymerization via 1,4-addition reaction to form an ene-yne alternating polymer chain. PDA-based two/three dimensional structures have been used for colorimetric or fluorescent devices, sensing biological as well as chemical components. This color-generating material also present a very high charge carrier mobility, allowing its application as field-effect transistor (FET). The immobilized PDA structures or films have distinct advantages for the detection of low concentration target molecules over the aqueous solution-based detection systems. In the present review, reported detection methods by using various PDA structures are summarized with updated references.