• Title/Summary/Keyword: Brn-3a

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Construction of a High-Quality Yeast Two-Hybrid Library and Its Application in Identification of Interacting Proteins with Brn1 in Curvularia lunata

  • Gao, Jin-Xin;Jing, Jing;Yu, Chuan-Jin;Chen, Jie
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.108-114
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    • 2015
  • Curvularia lunata is an important maize foliar fungal pathogen that distributes widely in maize growing area in China, and several key pathogenic factors have been isolated. An yeast two-hybrid (Y2H) library is a very useful platform to further unravel novel pathogenic factors in C. lunata. To construct a high-quality full length-expression cDNA library from the C. lunata for application to pathogenesis-related protein-protein interaction screening, total RNA was extracted. The SMART (Switching Mechanism At 5' end of the RNA Transcript) technique was used for cDNA synthesis. Double-stranded cDNA was ligated into the pGADT7-Rec vector with Herring Testes Carrier DNA using homologous recombination method. The ligation mixture was transformed into competent yeast AH109 cells to construct the primary cDNA library. Eventually, a high qualitative library was successfully established according to an evaluation on quality. The transformation efficiency was about $6.39{\times}10^5$ transformants/$3{\mu}g$ pGADT7-Rec. The titer of the primary cDNA library was $2.5{\times}10^8cfu/mL$. The numbers for the cDNA library was $2.46{\times}10^5$. Randomly picked clones show that the recombination rate was 88.24%. Gel electrophoresis results indicated that the fragments ranged from 0.4 kb to 3.0 kb. Melanin synthesis protein Brn1 (1,3,8-hydroxynaphthalene reductase) was used as a "bait" to test the sufficiency of the Y2H library. As a result, a cDNA clone encoding VelB protein that was known to be involved in the regulation of diverse cellular processes, including control of secondary metabolism containing melanin and toxin production in many filamentous fungi was identified. Further study on the exact role of the VelB gene is underway.

Efficient Mechanism for receiver and sink node in Wireless Sensor Networks (무선 센서 네트워크에서 수신 및 싱크 노드를 위한 효율적인 데이터 전송 방법)

  • Jeon, JunHeon
    • The Journal of Korean Association of Computer Education
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    • v.23 no.3
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    • pp.65-70
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    • 2020
  • In wireless sensor networks, data packets are send to the sink node. So traffic increases near the sink node. This causes delay and collision. To solve this problem, the proposed mechanism used half rotation antenna. By using a half rotation antenna, the delay of data packets can be reduced. Also we propose a method to efficiently use the energy of the node using BRN(backup receiver node) and increase the lifetime of the entire networks. Our numerical analysis and simulation results show that our mechanism outperforms RI-MAC protocol in terms of energy consumption and transmission delay.

Combining Ginsenoside F1 with (-)-Epigallocatechin Gallate Synergistically Protects Human HaCaT Keratinocytes from Ultraviolet B-Induced Apoptosis (Ginsenosdie F1과 EGCG의 상승작용에 의한 자외선조사에 의한 세포 사멸 방지)

  • Tae Ryong, Lee;Si Young, Cho;Eun Hee, Lee;Myeong Hoon, Yeom;Ih-Seop, Chang
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.30 no.2
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    • pp.253-261
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    • 2004
  • Ginsenosides and green tea extracts show a variety of biomedical efficacies such as anti-aging, anti-oxidation and anti-tumor-promotion effects. (-)-Epigallocatechin-3-gallate (EGCG) has been reported to inhibit the UVB-induced apoptosis by increasing the Bcl-2-to-Bax ratio. We have previously shown that ginsenoside Fl protects human HaCaT cells from ultraviolet-B (UVB)-induced apoptosis by maintaining constant levels of Bcl-2 and Brn-3a. Here, we investigate the combined effect of ginsenoside Fl and EGCG on the protection of human HaCaT keratinocyte against UVB-induced apoptosis. When treated individually, although 5 ${\mu}$M ginsenoside Fl and 50${\mu}$M EGCG protected cells from UVB-induced apoptosis, 2${\mu}$M ginsenoside Fl or 10${\mu}$M EGCG treatment showed very little protection effect. However, cotreatement of 2${\mu}$M ginsenoside Fl and 10${\mu}$M EGCG successfully protected HaCaT cells from UVB-induced cell death. As expected, combining ginsenoside Fl and EGCG efficiently prevented UVB-induced decrease of Bcl-2 and Brn-3a expression. In addition, cotreatment with ginsenoside F1 and EGCG prevented the dephosphorylation of Rb, whereas individual treatment with ginsenoside Fl or EGCG failed to prevent the dephosphorylation of Rb even at high concentrations.

Regenerative Effect of Adipose Derived Mesenchymal Stem Cells on Ganglion Cells in the Hypoxic Organotypic Retina Culture

  • Meital Ben Dov;Bryan Krief;Moshe Benhamou;Ainat Klein;Shula Schwartz;Anat Loewenstein;Adiel Barak;Aya Barzelay
    • International Journal of Stem Cells
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    • v.16 no.2
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    • pp.244-249
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    • 2023
  • Background and Objectives: To examine whether ischemic retinal ganglion cells (RGCs) will be salvaged from cell death by human adipose-derived mesenchymal stem cells (ADSCs) in an organotypic retina model. Methods and Results: Deprived of arterial oxygen supply, whole mice retinas were cultured as an ex vivo organotypic cultures on an insert membrane in a 24-well plate. The therapeutic potential of ADSCs was examined by co-culture with organotypic retinas. ADSCs were seeded on top of the RGCs allowing direct contact, or at the bottom of the well, sharing the same culture media and allowing a paracrine activity. The number of surviving RGCs was assessed using Brn3a staining and confocal microscopy. Cytokine secretion of ADSCs to medium was analyzed by cytokine array. When co-cultured with ADSCs, the number of surviving RGCs was similarly significantly higher in both treatment groups compared to controls. Analysis of ADSCs cytokines secretion profile, showed secretion of anti-apoptotic and pro-proliferative cytokines (threshold>1.4). Transplantation of ADSCs in a co-culture system with organotypic ischemic retinas resulted in RGCs recovery. Since there was no advantage to direct contact of ADSCs with RGCs, the beneficial effect seen may be related to paracrine activity of ADSCs. Conclusions: These data correlated with secretion profile of ADSCs' anti-apoptotic and pro-proliferative cytokines.

Intraocular Pressure and Neuroprotective Effect of water and ethanol extracts from Leaves of Acer palmatum thumb. of glaucoma in DBA2/J mice (녹내장 동물모델에서 단풍나무 잎 열수 및 에탄올 추출물의 안압 및 신경세포 보호 효과)

  • Oh, Tae Woo;Park, Kwang-Il;Ma, Jin Yeul
    • Herbal Formula Science
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    • v.29 no.2
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    • pp.93-103
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    • 2021
  • Objectives : This study was conducted to evaluate the Intraocular pressure (IOP) and retina neuroprotective effect of Acer palmatum thumb. extract in glaucoma of DBA2/J mice vivo model. Methods : The amount of change in intraocular pressure(IOP) was measured every two weeks for 30 weeks in the DBA2/J glaucoma animal model. The increased IOP were regrouping into drug treatment groups. EW and EE (1, 2 mg/㎖)extract was eye drop administered twice per day for 8 weeks. After administration of the extract, IOP was measured every 1 week at a fixed time for 8 weeks, and protein expression levels and serological changes related to retinal function were investigated. Results : The DBA2/J mice were IOP increased as time dependent, resulting in damage to the retinal function protein related in glaucoma animal model. However, administration of extracts from Leaves of Acer palmatum thumb. prevented an increase IOP, increased for Brn3α, PKCα, and OPN1SW. Additionally, the experssion of Aterocyte maker, GFAP was decreased in DBA2/J mice. Conclusions : These results demonstrate that both IOP and age can be considered in studies using DBA2/J mice and suggest that the extracts from Leaves of Acer palmatum thumb. regulating of IOP and expression of proteins related to retinal function.