• Title/Summary/Keyword: ${\beta}-galactosidase$ assay

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Comparative Study of Antimicrobial and Cytotoxic Effects of 1, 2-Octanediol and 1, 2-Octanediol Galactoside (1, 2-Octanediol과 1, 2-Octanediol Galactoside의 항균력 및 세포독성 비교연구)

  • Kim, Jun-Sub;Jin, Hong-Jong;Jung, Kyung-Hwan
    • Journal of the Korean Applied Science and Technology
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    • v.38 no.3
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    • pp.629-637
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    • 2021
  • To develop a safer cosmetic preservative, we carried out a comparative study on characteristics of OD and OD-gal, where OD-gal was synthesized from OD using E. coli β-gal. OD-gal synthesis was confirmed by mass spectrometry analysis as sodium adduct ion (m/z=331.1731) and protonated ion (m/z=309.1926) of OD-gal. To compare the antimicrobial activities of OD and newly synthesized OD-gal, MIC values were investigated using E. coli, S. aureus, C. albicans, and A. niger. As a result, it was observed that there was no remarkable difference between MIC values of OD and OD-gal. In addition, to compare the cytotoxicity of OD-gal and OD, HaCaT cells were treated with OD or OD-gal, and then cell viability was quantified using EZ-Cytox assay. In the case of 1.5% OD, the cell viability was 64% at 24 h and 42% at 48 h compared to the control, and cell viability of 1.5% OD-gal-treated cells showed no significant change at 24 h and was 85% at 48 h. Consequently, the cytotoxicity of OD-gal-treated cells was reduced by more than 40% when compared with that of OD-treated cells. Thus, the newly synthesized OD-gal in this study is safer than the existing OD used as a cosmetic additive. In the future, OD-gal will be applicable as a substitute for OD as a less toxic preservative for the cosmetic industry.

Antioxidant and Anti-aging Effects of Extracts from Leaves of the Quercusaliena Blume on Human Dermal Fibroblast (피부 섬유아세포에서 갈참나무 잎 추출물의 항산화 및 항노화 효능)

  • Choi, Sun-Il;Lee, Jong Seok;Lee, Sarah;Yeo, Joohong;Jung, Tae-Dong;Cho, Bong-Yeon;Choi, Seung-Hyun;Sim, Wan-Sup;Han, Xionggao;Lee, Jin-Ha;Kim, Jong Dai;Lee, Ok-Hwan
    • Journal of Food Hygiene and Safety
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    • v.33 no.2
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    • pp.140-145
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    • 2018
  • The skin of the human body occupies the largest surface area of the body and acts as a protection for the person's internal organs. As such, the skin is a major target of oxidative stressors, and these oxidative stressors are known to contribute to skin aging over the course of time. For the most part, an antioxidant is an effective approach to utilize to prevent symptoms related to the reactive oxygen species (ROS)-induced aging of the skin. Therefore, we investigated the antioxidant and anti-aging activity of the leaves of the Quercusaliena Blume extract (QBE). In our study, we confirmed that the cell viability tested with XTT {2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide innersalt} assay was not affected up to a concentration of $100{\mu}g/mL$. In addition, the cell viability of HDF cells induced by hydrogen peroxide was recovered from 81% to 104% after treatment with QBE, which showed the greater protective effect than that of ascorbic acid. Treatments of QBE dose-dependently inhibited reactive oxygen species (ROS) production in HDF cells induced by hydrogen peroxide, which correlated with their protective effects on cell viability. Since QBE treatment exhibited the suppression effect of skin aging by decreasing the ROS production, QBE could be used as a not only natural anti-aging but also antioxidant resource.

SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT (인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구)

  • Choi, Won-Jae;Ahn, Kang-Min;Hwang, Soon-Jeong;Choung, Pill-Hoon;Kim, Myung-Jin;Kim, Nam-Yeol;Yoo, Sang-Bae;Jahng, Jeong-Won;Kim, Hyun-Man;Kim, Joong-Soo;Kim, Yun-Hee;Kim, Soung-Min;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.3
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.