• Polymer(Korea)
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• v.38 no.1
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• pp.54-61
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• 2014

Water-soluble sulfobetaine chitosan (SCs) was prepared for a blending film with Bombyx mori silk fibroin (SF) by reacting chitosan with 1,3-propanesultone. A series of SF/SCs blended films were successfully prepared by mixing aqueous solutions of B. mori SF and SCs. The SF/SCs blended films were examined through spectroscopic and thermal analysis to determine the morphological changes of SF in the SCs. The effects of the SF/SCs blend ratios on physical and mechanical properties were investigated to discover the feasibility of using these films as biomedical materials such as artificial skin and wound dressing. X-ray analysis showed good compatibility between the two biopolymers. The in vitro degradation behavior of the SF/SCs blended films was systematically investigated for up to 8 weeks in phosphate buffered saline solution at $37^{\circ}C$ and showed a mass loss of 46.4% after 8 weeks. All films showed no cytotoxicity by MC3T3-E1 assay. After 3 days of culture, the relative cell number on all the SF/SCs films was slightly lower than that of an optimized tissue culture plastic.

• Polymer(Korea)
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• v.32 no.5
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• pp.403-408
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• 2008

Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. Keratin is useful as natural protein. We developed the keratin loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds (keratin/PLGA) for the possibility of the application of the tissue engineering using bone marrow mesenchymal (BMSCs). Keratin/PLGA (contents 0%, 10%, 20% and 50% of PLGA weight) scaffolds were prepared by solvent casting/salt leaching method. We characterized porosity, wettability, and water uptake ability, DSC of keratin/PLGA scaffold. We seeded BMSCs isolated from the femurs of rat into the inner core of the hybrid scaffold. Celluar viability were assayed by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) test. We confirmed that keratin/PLGA scaffold is hydrophilic by wettability, and water uptake ability measurement results. In MTT assay results, cell viability in scaffolds impregnated 10 and 20 wt% of keratin were higher than other scaffolds. In conclusion, we suggest that keratin/PLGA scaffold may be useful to tissue engineering using BMSCs.

• The Journal of Korean Obstetrics and Gynecology
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• v.27 no.1
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• pp.65-80
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• 2014

Objectives: The object of this study was to observe the in vitro antibacterial effects of Gagam-seopyoungjeon aqueous extracts (GGSYJ) against Gardnerella vaginalis and the possible synergic combination effects with clindamycin. Methods: Antibacterial activities against Gardnerella vaginalis of GGSYJ were detected using minimal inhibition concentration (MIC), and the effects on the bacterial growth curve were also monitored at MIC and MIC${\times}$2 levels. The combination effects of GGSYJ with clindamycin were observed by checkboard microtiter assay, and the effects of bacterial growth curve treated with GGSYJ MIC+clindamycin MIC, 1/2 MIC and 1/4 MIC, respectively. The effects on the bacterial invasion and intracellular killing of GGSYJ were also observed using human vaginal epithelial (VK2) and murine macrophage (Raw264.7) cells with combination effects with clindamycin after treatment of GGSYJ MIC+clindamycin 1/2 MIC, 1/4 MIC and 1/6 MIC, respectively. Results: The MIC of clindamycin and GGSYJ against Gardnerella vaginalis were detected as $0.012{\pm}0.006$ (0.004~0.016)${\mu}g/ml$ and $1.016{\pm}0.524$ (0.391~1.563) mg/ml, respectively. Clindamycin and GGSYJ were also showed marked dosage-dependent inhibition of bacterial growth, and significant decreases of viable cells were detected in clindamycin MIC+GGSYJ MIC and clindamycin 1/2 MIC+GGSYJ MIC treatment as compared with each of single clindamycin MIC and GGSYJ MIC treatments. And significant decreases of intraepithelial and intra-macrophage viable bacteria numbers were detected in clindamycin 1/2 MIC+GGSYJ 1/2 MIC and clindamycin 1/4 MIC+GGSYJ 1/2 MIC treatment as compared with each of single clindamycin GGSYJ 1/2 MIC treatments, respectively. Conclusions: GGSYJ showed slight antibacterial effects against Gardnerella vaginalis, but they showed dosage-dependent inhibitory effects on the bacterial growth and VK2 epithelial invasions of bacteria with favorable accelerating effects of intracellular killing activities of macrophages. In addition, combination of GGSYJ also increased the inhibitory effects of clindamycin on the epithelial invasions of Gardnerella vaginalis and intracellular killing activities of macrophages against Gardnerella vaginalis as 2-fold higher as compared with clindamycin single treatment, respectively. Therefore, we expected that the clinical dosages of clindamycin can be reduced as 1/2 levels as combination with GGSYJ.

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.161-165
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• 2012

Antimicrobial peptides of insects are found and reported as immune defence system against infectious agents. The peptides are produced by fat body cells and thrombocytoids, a blood cell type. Defensin is 38-45 amino acids long and consists of an ${\alpha}$-helix linked by a loop to an antiparallel ${\beta}$-sheet. Defensin from a bumblebee, Bombus ignitus, is known to comprise 52 amino acid residues. This peptide consists of two ${\alpha}$-helixes; ACAANCLSM and KTNFKDLWDKRF and one ${\beta}$-sheet; GGRCENGVCLCR. We carried out antibacterial activity test by radial diffusion assay against Staphylococcus aureus (Gram positive), Escherichia coli (Gram negative), Pseudomonas syringae (Gram negative), Candida albicans (fungi), MDRPA, MRSA, and VRE (antimicrobial resistant microbes) with synthetic oligopeptides from Peptron (Daejeon, Korea). The predicted curtailment fragment (GGRCEVCLCR-$NH_2$) for ${\beta}$-sheet had strong antibacterial activity when internal amino acids were removed. But, curtailment fragments (ACAANCLSM-$NH_2$ and TNFKDLWDKR-$NH_2$) of ${\alpha}$-helix were not showed antibacterial activity. These synthetic oligopeptides were showed the great activity against Gram positive and negative bacteria.

• Molecules and Cells
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• v.44 no.10
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• pp.770-779
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• 2021

Transgenic Arabidopsis thaliana expressing an anti-rabies monoclonal antibody (mAb), SO57, was obtained using Agrobacterium-mediated floral dip transformation. The endoplasmic reticulum (ER) retention signal Lys-Asp-Glu-Leu (KDEL) was tagged to the C-terminus of the anti-rabies mAb heavy chain to localize the mAb to the ER and enhance its accumulation. When the inaccurately folded proteins accumulated in the ER exceed its storage capacity, it results in stress that can affect plant development and growth. We generated T₁ transformants and obtained homozygous T₃ seeds from transgenic Arabidopsis to investigate the effect of KDEL on plant growth. The germination rate did not significantly differ between plants expressing mAb SO57 without KDEL (SO plant) and mAb SO57 with KDEL (SOK plant). The primary roots of SOK agar media grown plants were slightly shorter than those of SO plants. Transcriptomic analysis showed that expression of all 11 ER stress-related genes were not significantly changed in SOK plants relative to SO plants. SOK plants showed approximately three-fold higher mAb expression levels than those of SO plants. Consequently, the purified mAb amount per unit of SOK plant biomass was approximately three times higher than that of SO plants. A neutralization assay revealed that both plants exhibited efficient rapid fluorescent focus inhibition test values against the rabies virus relative to commercially available human rabies immunoglobulins. KDEL did not upregulate ER stress-related genes; therefore, the enhanced production of the mAb did not affect plant growth. Thus, KDEL fusion is recommended for enhancing mAb production in plant systems.

• Korean Journal of Organic Agriculture
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• v.28 no.4
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• pp.659-677
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• 2020

Leonurus japonicus (L. japonicus) Houtt., a biennial plant in the Lamiaceae family is broadly distributed in Asia such as Korea, China, Japan. The aerial part of L. japonicus is used as a traditional medicine to treat uterine disease including dysmenorrhea, amenorrhea, sterility. In this study, we examined the antioxidant and anti-inflammatory effects of L. japonicus ethanol extracts. The antioxidant activity of L. japonicus was measured by total polyphenol and flavonoid content, and DPPH, ABTS scavenging, reducing power activity, and intracellular ROS expression assay. The anti-inflammatory effects were measured by nitric oxide (NO), cytokines (TNF-α and IL-1β) production and inflammatory protein expression in LPS-induced RAW 264.7 cells. Total polyphenol and flavonoid content of L. japonicus were 51.40 ± 0.47 mg of gallic acid equivalents/g and 73.28 ± 0.10 mg of rutin equivalents/g respectively. DPPH, ABTS radical scavenging activity and reducing power activity tended to increase concentration-dependent and treatment L. japonicus with 400 ㎍/mL reduced ROS production by 69.5%. Furthermore, L. japonicus inhibited NO, TNF-α and IL-1β production in a concentration-dependant manner and reduced the expression levels of inflammatory proteins via regulating NF-κB, MAPK pathway. Therefore, we suggest that L. japonicus could be a natural antioxidants and medicinal source to treat oxidative stress and inflammation-related disease.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.20 no.6
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• pp.430-438
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• 2019

This study is to investigate increase immunomodulatory activity of Chungkukjang added with Dendropanax morbifera extract. There are four groups divided that Control, Dendropanax morbifera extract group, Chungkookjang group and Chungkukjang added to Dendropanax morbifera extract group which of five mouses each group. Their immunological characteristics were compared with either general Chungkukjang or non treated group in the tested male ICR mice. There were no significant difference in body weight and organ weight. In the plaque forming cell assay, Dendropanax morbifera extract 250 and 500 mg/kg groups measured significant increase over the control group. The values of lymphocytes were significantly increased in the Dendropanax morbifera extract 500 mg/kg group campared with the control group. In the general Chungkukjang 400 mg/kg added with Dendropanax morbifera extract 500 mg/kg group, total serum immunoglobulin G concentration was significantly higher than the control group and their spleen tissues observed proliferation of white puls. These results demonstrated that Chungkukjang Added with Dendropanax morbifera extract was provided enhance of immunomodulatory activity and suggest that it can be used as various functional foods, based on foods promote immune system health.

• Journal of Life Science
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• v.28 no.12
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• pp.1432-1437
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• 2018

Clara cell secretory protein (CCSP) plays an important role in protecting the lungs from inflammation. This research focuses on identifying the cis-element for binding the repressor of CCSP gene expression. A DNase I footprinting experiment revealed three protected regions between -812 and -768 bp (45 bp) of the mCCSP promoter. One motif (D3: GCCTGGGAA) was 100% conserved across rat, hamster, and human. The addition of excess amounts of the D3 motif exhibited high competition within that 45 bp range in an electrophoretic mobility shift assay. However, when mutated D3 ($G{\underline{AA}}TG{\underline{TT}}AA$) was used, the competition was significantly reduced. This demonstrates that the D3 motif within that 45 bp region of the mCCSP promoter is an important site for the protein-DNA interaction. Transient transfection assays with -756 Luc resulted in highly decreased expression of CCSP than those with -812 Luc, suggesting that the 45 bp could function as a binding site for the repressor. Co-transfection of KLF4 exhibited significant repression of the -812 Luc but not the -768 Luc which clearly shows that KLF4 might function as a repressor for the CCSP gene and also suggests that the D3 motif is strongly involved in the binding of KLF4. In addition, when anti-KLF4 antibody was added, super-shifted bands were observed. This result demonstrates that KLF4 could function as a repressor by binding to this 45 bp region of the CCSP promoter and that the D3 motif might be involved in the specific binding of KLF4.

• Journal of the Korean Applied Science and Technology
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• v.36 no.2
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• pp.488-497
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• 2019

The purpose of this study was to confirmed anti-inflammatory effect the apple Induced by mutants with ${\gamma}-Ray$ extract. Cell viability was assessed by MTT assay using RAW 264.7 cells. The extracts measured through changes in the levels of reactive oxygen species (ROS), nitric oxide (NO), inflammatory cytokines, NF-kB, and COX-2 on LPS-induced RAW 264.7 cells. All test results were analyzed by ELISA reader, Luminex and RT-PCR. In result, the extracts was not toxic below in 25 ug/ml, and extracts was inhibited the productions nitric oxide, ROS, cytokines (IL-1b, IL-6, TNF-a), NF-kB and COX-2 in LPS-induced RAW 264.7 cells. Also, the expression levels were decreased on mRNA of $NF-{\kappa}B$ and COX-2. In other words, Perilla frutescens var. crispa Induced by mutants with ${\gamma}-Ray$ extracts showed significant anti-inflammatory effect. These results may be developed as a raw material for new health food and therapeutics to ease the related to the above mediators.

• Journal of Life Science
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• v.29 no.9
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• pp.972-976
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• 2019

Polyopes affinis is a kind of red algae found in the South coast and near Jeju Island of Korea. The purpose of this study was to investigate the effects of Polyopes affinis ethanol extract (PAEE) on melanogenesis in ${\alpha}-MSH$ stimulated B16F10 melanoma cells. Melanoma cells were cultured for 72 hr treated with PAEE. Total melanin content and the activity of tyrosinase, a key enzyme in melanogenesis, were measured. When the melanin content in B16F10 melanoma cells was tested, PAEE was decreased in a dose-dependent manner: treatment with 100, 300, and $500{\mu}g/ml$ caused 25%, 30%, and 35% reduction, respectively. Treatment of 100, 300, and $500{\mu}g/ml$ of PAEE caused 6%, 12%, and 21% reduction of tyrosinase activities in B16F10 melanoma cells. Also, PAEE suppressed the expression of tyrosinase, tyrosinase-related protein-1, tyrosinase-related protein-2, and melanocyte-inducing transcription factor in B16F10 melanoma cells. A concentration of $500{\mu}g/ml$ of PAEE showed a greater decrease in tyrosinase activity, melanin content, and melanogenic enzyme protein expression. These results indicate that PAEE inhibits melanin synthesis and tyrosinase activity, and Polyopes affinis ethanol extract could be used as a functional whitening agent.