• Title/Summary/Keyword: 세포융합기술

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Expression of Recombinant Human Epidermal Growth Factor as a Active Form through Codon Optimization with E. coli and Co-expression of Chaperone (코돈 최적화 및 샤페론 공발현을 통한 활성 형태의 재조합 인간 상피세포성장인자의 발현)

  • Jang, Eun-Bin;Kim, Jun Su;Lee, Woo-Yiel
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.9
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    • pp.559-568
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    • 2020
  • Epidermal growth factor (EGF) is a hormone protein that affects cell growth and proliferation, and has various medical applications. In the present study, the gene of human EGF was codon-optimized with E. coli and the expression vector was constructed by cloning into pRSET. In order to obtain the recombinant human EGF in an active form rather than an inclusion body, chaperone co-expression was attempted along with codon optimization, for the first time. The expressed human EGF was isolated in the pure form by performing Ion Exchange Chromatography in two consecutive runs. ELISA analysis showed that the activity of purified EGF was greater than 99%, which is similar to commercially available EGF. Cell proliferation test confirmed that the recombinant human EGF has the ability to promote cell proliferation of human skin fibroblasts. The human EGF expression system of this study gives a significant amount of protein, and does not require the renaturation step and the additional chromatographic system to remove a fusion contaminant, thereby providing a very useful alternative to conventional expression systems for the preparation of recombinant human EGF.

The Effects of Anti-Inflammatory Activities and Active Fractions Analysis of Ethanol Extract from Red Rose Petals (붉은 장미꽃잎 에탄올 추출물의 활성 분획물 분석 및 항염증 활성 효과)

  • Kim, Hyun Kyoung
    • The Journal of the Convergence on Culture Technology
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    • v.6 no.2
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    • pp.543-551
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    • 2020
  • Red rose petals are usually disposed but they are an abundant source of phenolics and traditionally used as food supplement and as herbal medicine. Of the Various phenolics, they are known to have anticancer, antioxidant, and anti-inflammatory properties. In this study, we investigated the anti-inflammatory effects of red rose ethanolic extracts(GRP) on lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results demonstrated that pretreatment of GRP(500㎍/mL) significantly reduced NO production by suppressing iNOS protein expression in LPS-stimulated cells. Anti-inflammatory effects byred rose petal were observed in the following. Red rose petal inhibited the translocation of NF-κB from the cytosol to the nucleus via the suppression of IκB-α phosphorylation and also inhibited LPS-stimulated NF-κB transcriptional activity. These findings suggest that red rose petal exert anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of red rose petal. Therefore, red rose petal could be regarded as a potential source of natural anti-inflammatory agents.

Antioxidant Activity of Native Korean Halophyte Extracts and Their Anti-biofilmActivity against Acinetobacter baumannii (한국 자생 염생식물 추출물의 항산화 활성 및 다재내성 Acinetobacter baumannii에 대한 항생물막 활성)

  • Eun Seong Lee;Jeong Woo Park;Ki Hwan Moon;Youngwan Seo
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1015-1024
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    • 2023
  • Antibiotics have greatly contributed to the treatment and prevention of bacterial diseases in humans, animals, and fish. However, antibiotic misuse has led to the emergence and spread of multidrug-resistant bacteria. In addition to antibiotic discovery research, efforts are being made to combat such multidrug-resistant bacteria using antimicrobial agents, antioxidants, host immune enhancement, probiotics, and bacteriophages, as well as various symptomatic therapies. To discover novel bioactive compounds, it is crucial to adopt approaches that incorporate fresh ideas, new targets, innovative techniques, and untapped resources. Halophytes are plants that grow in high-salt soils and are known to adapt to salt-induced stress through unique metabolic processes that produce secondary metabolites. This study aimed to investigate the effects of extracts of halophytes native to Korea on oxidative stress and to determine whether they exert inhibitory activity against biofilms, which are major pathogenic factors of infectious bacteria. The Acinetobacter baumannii strain ATCC 17978, a representative drug-resistant bacterium, was used to measure anti-biofilm activity. The results showed that Aster spathulifolius, Carex kobomugi, Rosa rugosa, and Asparagus cochinchiensis exerted strong antioxidant and anti-biofilm effects without affecting bacterial growth itself. The halophytes used in this study are promising candidates for the development of pharmaceutical agents with antioxidant and antimicrobial properties.

Counting Harmful Aquatic Organisms in Ballast Water through Image Processing (이미지처리를 통한 선박평형수 내 유해수중생물 개체수 측정)

  • Ha, Ji-Hun;Im, Hyo-Hyuk;Kim, Yong-Hyuk
    • Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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    • v.6 no.3
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    • pp.383-391
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    • 2016
  • Ballast water provides stability and manoeuvrability to a ship. Foreign harmful aquatic organisms, which were transferred by ballast water, cause disturbing ecosystem. In order to minimize transference of foreign harmful aquatic organisms, IMO(International Maritime Organization) adopted the International Convention for the Control and Management of Ship's Ballast Water and Sediments in 2004. If the convention take effect, a port authority might need to check that ballast water is properly disposed of. In this paper, we propose a method of counting harmful aquatic organisms in ballast water thorough image processing. We extracted three samples from the ballast water that had been collected at Busan port in Korea. Then we made three grey-scale images from each sample as experimental data. We made a comparison between the proposed method and CellProfiler which is a well known cell-counting program based on image processing. Setting of CellProfiler is empirically chosen from the result of cell count by an expert. After finding a proper threshold for each image at which the result is similar to that of CellProfiler, we used the average value as the final threshold. Our experimental results showed that the proposed method is simple but about ten times faster than CellProfiler without loss of the output quality.

Inhibitory effects of ethanol extract from Vicia amoena on LPS(Lipopolysaccharide) induced nitric oxide and prostaglandin E2 production in RAW264.7 macrophage cell (갈퀴나물 에탄올 추출물의 RAW264.7 대식세포에서 LPS(Lipopolysaccharide)로 유도된 nitric oxide 및 prostaglandin E2 생성 저해효과)

  • Nam, Jung-Hwan;Park, Soo-Jin
    • Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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    • v.9 no.6
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    • pp.443-450
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    • 2019
  • Vicia amoena has traditionally been used to treat disease of rheumatism, arthralgia, muscular paralysis, abscess and eczema, and it has anti-inflammatory properties. However, validity of the anti-inflammatory activity has not been scientifically in vestige acted so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of V. amoena using the ethanolic extract. To evaluate the anti-inflammatory effects, we examined the inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2) on RAW264.7 cells. Our results indicated that ethanolic extract of V. amoena significantly inhibited the LPS-induced NO and PGE2 production in RAW264.7 cells. The ethanolic extract of V. amoena has inhibited the PGE2 production by 88.0±0.8 % at the concentration of 40㎍/ml. This results showed that ethanol extract of V. amoena is expected to be a good candidate for development into source of inflammation inhibitor

Effect of Cnidium japonicum Miq. Crude Extracts on UVB-induced Photoaging Damage in Human Keratinocytes (HaCaT 세포에서 UVB로 유도된 광노화에 대한 갯사상자 추출물의 효능)

  • Eun Seong Lee;Jung Hwan Oh;Chang-Suk Kong;Youngwan Seo
    • Journal of Life Science
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    • v.33 no.5
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    • pp.414-421
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    • 2023
  • Cnidium japonicum (C. japonicum) is a type of halophyte that inhabits soil of a high salinity, and according to previous studies, it is known to have antitumor effects. However, the skin's protective effect, particularly against UVB irradiation, has not been revealed. In this study, C. japonicum crude extract was studied to determine its effect on damage to human keratinocytes (HaCaT) induced by UVB irradiation, and ROS assays were performed, the results of which showed that C. japonicum crude extract affects UVB-induced photoaging damage in human keratinocytes. To examine inhibitory effects against the expressions of MMPs, RT-PCR and Western blot assay were performed by treating the crude extract at concentrations of 10, 50, and 100 ㎍/ml by irradiating UVB at 15 mJ/cm2. As a result, it was confirmed that the mRNA and protein expression levels of MMP-1, MMP-3, and MMP-9 decreased in the group treated with C. japonicum crude extract, which also effectively regulated the antioxidant defense mechanism pathway by activating JNK, ERK, and p38. In conclusion, the current study suggested the possibility that C. japonicum could be used as a raw material for anti-photoaging cosmeceuticals in the future.

The Anti-Bacterial Activity and Anti-Inflammatory Effect of Ethanol Complex Extracts of Safflower and Mother Wort (사플라워와 마더워트 등 에탄올복합추출물의 항균활성과 항염증 효과)

  • Hyun Kyoung Kim;Yungi Lee;Subin Choi;DO Wan Kim
    • The Journal of the Convergence on Culture Technology
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    • v.9 no.1
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    • pp.717-724
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    • 2023
  • we are investigated the anti-inflammatory effects of Safflower and Mother wort Ethanol Complex Extracts(SEC) on lipopolysaccharide (LPS)-activated RAW 264.7 cells. The results demonstrated that pretreatment of SEC(500㎍/mL) significantly reduced NO production by suppressing iNOS protein expression in LPS-stimulated cells. Anti-inflammatory effects by Safflower and Mother wort Ethanol Complex Extracts were observed in the following. Safflower and Mother wort Ethanol Complex Extracts inhibited the translocation of NF-κB from the cytosol to the nucleus via the suppression of IκB-α phosphorylation and also inhibited LPS-stimulated NF-κB transcriptional activity. These findings suggest that Safflower and Mother wort Ethanol Complex Extracts exert anti-inflammatory actions and help to elucidate the mechanisms underlying the potential therapeutic values of Safflower and Mother wort Ethanol Complex Extracts. Therefore, Safflower and Mother wort Ethanol Complex Extracts could be regarded as a potential source of natural anti-inflammatory agents.

In-vitro Neuroprotective Effect of Aricumin(Turmeric extract) (아리큐민의 In-vitro 신경보호 효과)

  • Yoon, Nam kyu;Kim, Byung Kwon;Ryu, Hyeon yeol;Seo, Bo Seung;Shin, Chang Ho;Kim, Kwan Kyu;Lee, Han Joo
    • The Journal of the Convergence on Culture Technology
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    • v.8 no.3
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    • pp.291-296
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    • 2022
  • This study was conducted on curcumin which had increased bioavailability as a potential AChE inhibitor for the treatment of neurodegenerative diseases. The purpose of this study is to confirm the in vitro neuroprotective effect on Aricumin (turmeric extract). To confirm the neuroprotective effect, AChE inhibition for Aricumin was evaluated, and cell viability was analyzed for HT-22cell, and oxidative stress (glutamate, H2O2)-induced HT-22 cytotoxicity was evaluated. As a result of the change in the AChE inhibition rate of Aricumin (Turmeric extract), it was confirmed that Aricumin at a concentration of 39.06㎍/ml or higher inhibited AChE activity by about 20% and more. And it was confirmed that the cytotoxicity of HT-22 cells induced by oxidative stress (Gluamate 5 mM and H2O2 500 µM) was significantly inhibited from 0.01 to 0.1 mg/ml concentration (p<005). These results suggest that Aricumin (turmeric extract) have potential neuroprotective effects.

Isolation and Fusion of Solanaceous Species Mesophyll Protoplast (가자과(茄子科)의 엽육조직(葉肉組織) 원형질체(原形質체体)의 분리(分離) 및 융합(融合))

  • Kwon, Oh Sung;Kim, Dal Dng
    • Current Research on Agriculture and Life Sciences
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    • v.2
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    • pp.15-22
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    • 1984
  • This study was conducted to identify the enzyme treatment time, enzyme concentration and plant growth condition for isolation of potato mesopyll, it was also performed to determine the adquate sucrose molarity on purification of protoplasts and to investigate the incubation time, PEG concentration and DMSO effect for potato-petunia protoplast fusion. The results were summarized as follows: The optimal time of incubation in enzyme solution was 3 - 4 hours and high humidity and low light intensity made plants more effective to protoplast releasing enzymes. Our experimental results showed that the pectolyase Y-23 was an ideal agent for isolation from mesophyll cultured in vitro compared with macerozyme. The enzyme solution with 0.5 % macerozyme and 2 % cellulase was very effective and the purity of healthy protoplast was better in 0.4 and 0.5 M sucrose than in others. It was revealed that the rate of potato-petunia fusion according to the incubation time with PEG was effective at 30 min incubation and percentage of protoplast aggregation was increased by high molecular weight and concentration of PEG. Percentage of potato-petunia protoplast heteroplasmic aggregation was increased by 4 to 16 % in PEG 6000 compared with PEG 4000 and PEG 1500. Addition of 5 to 10 % DMSO to the PEG solution increased to the heteroplasmic aggregation of potato-petunia from 2 to 4 %.

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Hormonal Study to Induce Direct Organ Differentiation of Kalanchoe pinnata by Tissue Culture (조직배양으로 Kalanchoe pinnata의 직접기관분화를 유도하기 위한 호르몬 연구)

  • Kim, Donggiun
    • The Journal of the Convergence on Culture Technology
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    • v.7 no.4
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    • pp.721-726
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    • 2021
  • Plant cells have a totipotencial capacity, the ability of each cell to produce a new complete individual through development. By applying this, several technologies are being developed for widespread application of somatic embryogenesis by processing hormones in vitro as a method of propagation of plants. In order to use this technology, in Kalanchoe pinnata, a plant capable of asexual reproduction with more regular cell division, kinetin belonging to cytokinin and picloram among hormones belonging to auxin were added in combination and treated for 8 weeks, and then the typical performance was evaluated. As a result of our experiment, the rooting effect in leaf slices showed a 70% incidence rate at a picloram concentration of 0.1 mg/L. It has been proven that a concentration difference of 1:5-1:10 in the ratio of kinetin and picloram is effective. It is the experimental result that the effect of auxin is essential for the development of Kalanchoe roots. As for the effect of shooting, the incidence rate was 60% at the picloram concentration of 0.5 mg/L. The kinetin concentration from 0.5 and 1.0 mg/L and has a significant effect on development. It has been proven that the ratio of kinetin to picloram is effective with a concentration difference of 1:1-1:2. These results show that the combination of cytokinin and auxin is crucially important for shooting. It is thought that it can be the basis of a technology for inducing mass proliferation in vitro by inducing direct organogenesis with a combination of hormones.