• Title/Summary/Keyword: in vitro/in vivo studies

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Biochemical and Molecular Biological Studies on the DNA Replication of Bacteriophage T7 (Bacteriophage T7의 유전자 복제기작에 관한 생화학적, 분자생물학적 특성 연구)

  • KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.28 no.2
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    • pp.209-218
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    • 1995
  • Bacteriophage T7 gene 2.5 protein, a single-stranded DNA binding protein, has been implicated in T7 DNA replication, recombination, and repair. Purified gene 2.5 protein has been shown to interact with the phage encoded gene 5 protein (DNA polymerase) and gene 4 proteins (helicase and primase) and stimulates their activities. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth. T7 phage that contain null mutants of gene 2.5 were constructed by homologous recombination. These mutant phage $(T7\Delta2.5)$ cannot grow in Escherichia coli. After infection of E. coli with $T7\Delta2.5$, host DNA synthesis is shut off, and $T7\Delta2.5$ DNA synthesis is reduced to less than $1\%$ of wild-type phage DNA synthesis (Kim and Richardson, 1993, Proc. Natl. Aca. Sci. USA, 90, 10173-10177). A truncated gene 2.5 protein $(GP2.5-\Delta21C)$ deleted the 21 carboxyl terminal amino acids was constructed by in vitro mutagenesis. $GP2.5-\Delta21C$ cannot substitute for wild-type gene 2.5 protein in vivo; the phage are not viable and exhibit less than $1\%$ of the DNA synthesis observed in wild-type phage-infected cells. $GP2.5-\Delta21C$ has been purified to apparent homogeneity from cells overexpressing its cloned gene. Purified $GP2.5-\Delta21C$ does not physically into「act with T1 gene 4 protein as measured by affinity chromatography and immunoblot analysis. The mutant protein cannot stimulate T7 gene 4 protein activity on RNA-primed DNA synthesis and primer synthesis. These results suggest that C-terminal domain of gene 2.5 protein is essential for protein-protein interactions.

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Artificial Vision Project by Micro-Bio Technologies

  • Kim Sung June;Jung Hum;Yu Young Suk;Yu Hyeong Gon;Cho Dong il;Lee Byeong Ho;Ku Yong Sook;Kim Eun Mi;Seo Jong Mo;Kim Hyo kyum;Kim Eui tae;Paik Seung June;Yoon Il Young
    • 한국가시화정보학회:학술대회논문집
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    • 2002.04a
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    • pp.51-78
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    • 2002
  • A number of research groups worldwide are studying electronic implants that can be mounted on retinal optic nerve/visual cortex to restore vision of patients suffering from retinal degeneration. The implants consist of a neural interface made of biocompatible materials, one or more integrated circuits for stimuli generation, a camera, an image processor, and a telemetric channel. The realization of these classes of neural prosthetic devices is largely due to the explosive development of micro- and nano-electronics technologies in the late $20^{th}$ century and biotechnologies more recently. Animal experiments showed promise and some human experiments are in progress to indicate that recognition of images can be obtained and improved over time. We, at NBS-ERC of SNU, have started our own retinal implant project in 2000. We have selected polyimide as the biomaterial for an epi-retinal stimulator. In-vitro and in-vivo biocompatibility studies have been performed on the electrode arrays. We have obtained good affinity to retinal pigment epithelial cells and no harmful effect. The implant also showed very good stability and safety in rabbit eye for 12 weeks. We have also demonstrated that through proper stimulation of inner retina, meaning vision can be obtained.

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Anti-cancer Properties and Relevant Mechanisms of Cordycepin, an Active Ingredient of the Insect Fungus Cordyceps spp., (동충하초 유래 cordycepin의 항암 활성 기전 최근 연구 동향)

  • Jeong, Jin-Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.25 no.5
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    • pp.607-614
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    • 2015
  • Cancers are the largest cause of mortality and morbidity all over the world. Cordycepin, an adenosine analog, is a major functional component of the Cordyceps species, which has been widely used in traditional Oriental medicine. Over the last decade, this compound has been reported to possess many pharmacological properties, such as an ability to enhance immune function, as well as anti-inflammatory, antioxidant and anti-cancer effects. Recently, numerous studies have reported interesting properties of cordycepin as a chemopreventive agent as well. There is an accumulating body of experimental evidences suggesting that cordycepin impedes cancer progression by promoting apoptosis, inducing cell cycle arrest, modulating intracellular signaling pathways, and inhibiting invasion and metastasis of cancer cells. In many cancer cell lines, cordycepin inhibits growth and cell cycle progression by inducing arrest of the G2/M phase, resulting from the inhibition of retinoblastoma protein phosphorylation and induction of cyclin-dependent kinase inhibitors. To induce apoptosis, cordycepin activates the extrinsic and intrinsic pathways, which promotes reactive oxygen species generation and the downstream activation of kinase cascades. Cordycepin also can activate alternative pathways to cell death such autophagy. In addition, cordycepin can inhibit the pro-metastatic processes of cancer cell detachment, migration, and invasion through a variety of mechanisms, including the nuclear factor-kappa B and activated protein-1 signaling pathways. In this review, we summarized the variety of action mechanisms by which cordycepin may mediate chemopreventive effects on cancer and discussed the potential of this natural product as a promising therapeutic inhibitor of cancer development.

A Modeling of Web-service for Construction CALS/EC Standard Guideline by using Component Based Development (컴포넌트 기반 개발방법론에 의한 건설 CALS/EC 표준지침 웹서비스의 모델링)

  • 이상호;정용환;김소운
    • Proceedings of the CALSEC Conference
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    • 2003.09a
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    • pp.78-83
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    • 2003
  • 본 연구에서는 건설CALS/EC 기반의 정보화 추진을 위한 정보시스템을 개발할 때 적용되는 기존의 소프트웨어 개발방법론의 문제점을 분석하고, 표준화되지 않은 개발방법론의 적용으로 시스템 개발에 비효율적인 비용을 투자하는 문제점과 특성상 입찰, 구매, 계약 등 분야별로 상이한 기능의 업무에 따라 구축되는 건설산업 정보화에 있어서 상호간에 데이터 및 프로세스를 원활히 통합하지 못하므로 인하여 발생되는 개발의 중복성, 정보 활용의 비효율성 등의 문제점을 개선하고자 하였다. 이를 위하여 최근 새로이 부각되고 있는 방법론인 컴포넌트 기반 개발방법론(CBD: Component-Based Development)을 사용하여 건선CALS/EC 표준지침 웹서비스를 위한 시스템의 업무프로세스를 모델링하여 사용자가 쉽게 재사용가능하고 타업무분야에 확대 가능한 방향을 제시하였다. 본 연구를 위하여 기존의 소프트웨어 개발 방법론의 적용상 문제점 분석을 통하여 컴포넌트 기반 개발방법론의 필요성을 증명하고 비즈니스 컴포넌트 프레임워크를 사용하여 건설 CALS/EC 표준지침 웹서비스의 업무프로세스에 적용할 기술적인 방법론을 고찰하고 업무에 효과적으로 적용할 수 있는 업무프로세스 컴포넌트를 제시하였다. 본 연구에서 도출된 컴포넌트 모델은 향후 타업무분야의 시스템을 개발할 때 사용자요구분석 단계부터 별도의 재 작업이 없이 사용될 수 있으며 시스템을 구현할 때 개발 모듈의 중복방지와 용이한 비즈니스로직의 변경 등이 가능하며, 추가의 업무 프로세스나 연관된 다른 분야의 업무프로세스의 반영 및 추가 시 컴포넌트의 활동모델을 쉽게 수정하여 정의함으로써 쉽게 시스템의 기능을 확장할 수 있다.LE 산정에 관한 지속적인 실험적 연구가 이루어져야 하겠다. 증가할 것이다. 또한 부분육을 이용한 완전제품, 적색육제품, 유기농이나 별미식 제품과 같은 형태의 다양한 포장육 제품이 도입 될 것으로 생각되어진다.e in vitro SPF test method will be able to be used as an alternative method for in vivo SPF in case of lotion and cream. replica. A statistically significant improvement of Star Fruit Leaf Extract BG30-treated site was seen in decreased wrinkles. Star Fruit Leaf Extract BG30 results in clinically visible improvement in wrinkling when used topically for 5 weeks. 또한 관계마케팅, CRM 등의 이론적 배경이 되고 있는 신뢰와 결속의 중요성이 재확인하는 결과도 의의라고 할 수 있다. 그리고 신뢰는 양사 간의 상호관계에서 조성될 수 있는 특성을 가진 반면, 결속은 계약관계 초기단계에서 성문화하고 규정화 할 수 있는 변수의 성격이 강하다고 할 수가 있다. 본 연구는 복잡한 기업간 관계를 지나치게 협력적 측면에서만 규명했기 때문에 많은 측면을 간과할 가능성이 있다. 또한 방법론적으로 일방향의 시각만을 고려했고, 횡단적 조사를 통하고 국내의 한 서비스제공업체와 관련이 있는 컨텐츠 공급파트너만의 시각을 검증했기 때문에 해석에서 유의할 필요가 있다. 또한 타당성확보 노력을 기하였지만 측정도구

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Effects of a Plant Water Extract Mixture(Ixeris sonchifolia Hance, Oenanthe javanica, Fagopyrum esculentum Moench, Hizikia fusiforme, Zingiber officinale Roscoe) on Mouse Immune Cell Activation (식물 혼합(고들빼기, 돌미나리, 메밀, 톳, 생강) 추출물이 마우스 면역 세포 활성에 미치는 영향)

  • Kim, Jung-Hee;Kim, Hyun-Sook;Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.20 no.1
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    • pp.74-78
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    • 2007
  • Ixeris sonchifolia Hance(Godulbaegi), Oenanthe javanica(Dolminari), Fagopyrum esculentum Moench(Buckwheat), Hizikia fusiforme(Seaweed Fusiforme) and Zingiber officinale Roscoe(Ginger) have all been used as one of the traditional remedies as well as food source. There are few studies However, on their immunomodulating effects have been reported. We previously reported that ex vivo supplementation of each of the Ish, Oj, Fem, Hf and Zor water extracts enhanced the splenocytes proliferation compared to the control group. In this study, the combined immunomodulative effects of a plant water extract mixture containing these five food sources(Ish+Oj+Fem+Hf+Zor) was compared to the individual effect of each. The production of cytokine(IL-1${\beta}$, IL-6, and TNF-${\alpha}$), secreted by macrophages stimulated with LPS or without, were detected via ELISA assay using a cytokine kit. After 48hrs of incubation with mitogen(ConA or LPS) stimulation, the mouse splenocyte proliferation in the experimental group had significantly increased at two different concentrations compared to the control group. The results of this study may suggest that the supplementing with a plant water extract mixture could regulate immune function by increasing splenocyte proliferation as well as enhance immune function by regulating the cytokine production capacity activated macrophages in mice.

Anti-oxidant Activities of Punica granatum Extracts (석류 추출물의 항산화 작용)

  • Roh, Bo-Kyung;Kim, Jee-Young;Kim, Ju-Yon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.2 s.51
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    • pp.207-212
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    • 2005
  • Pomegranate (Punica granatum L.) which is very rich in Polyphenols and tannins was recently reported its anti-oxidant activities and phytoestrogenic activities in vivo test and many clinical studies, but the effects of them on the skin have not been reported. The experiments were tarried out in vitro to determine anti-oxidant activities of pomegranate extracts on DPPH radical scavenging assay, NBT/Xanthine Oxidase-superoxide scavenging assay, silica-induced intracellular $H_2O_2$, hydroperoxide and superoxide generation assay in RAW 264.7 cells. It showed that the methanolic extract of dried pomegranate peels have the most significant anti-oxidant activities on free radical scavenging assay and inhibitory activities on silica-induced intracellular free radical generation in RAW 264.7 cells. The concentrated juice of pomegranate showed only DPPH radical scavenging activities and inhibited hyaluronidase activity. Moreover, pomegranate seed oil inhibited specially silica-induced intracellular hydroperoxide generation in RAW 264.7 cells. These results suggest that the methanolic extract of dried pomegranate peels and pomegranate seed oil have more anti-oxidant activity than concentrated juice of pomegranate. Thus the extracts of pomegranate peels and seed oil could be developed cosmetic ingredients for anti-aging.

Layered Double Hydroxide Nanoparticles for Bio-Imaging Applications (LDH 나노입자 기반의 바이오 이미징 소재)

  • Jin, Wenji;Ha, Seongjin;Lee, Dongki;Park, Dae-Hwan
    • Korean Chemical Engineering Research
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    • v.57 no.4
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    • pp.445-454
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    • 2019
  • Layered double hydroxides (LDHs) nanoparticles have emerged as novel nanomaterials for bio-imaging applications due to its unique layered structure, physicochemical properties, and good biocompatibility. Bio-imaging is one of the most important fields for medical applications in clinical diagnostics and therapeutics of various diseases. Enhanced diagnostic techniques are needed to realize new paradigm for next-generation personalized medicine through nanoscale materials. When nanotechnology is introduced into bio-imaging system, nanoparticle probes can endow imaging techniques with enhanced ability to obtain information about biological system at the molecular level. In this review, we summarize structural features of LDH nanoparticles with current issues of bio-imaging system. LDH nanoparticle probes are also discussed through in vitro as well as in vivo studies in various bio-imaging techniques including fluorescence imaging, magnetic resonance imaging (MRI), positron emission tomography (PET), and computed X-ray tomography (CT), which will have the potential in the development of the advanced nanoparticles with high sensitivity and selectivity.

Enhanced γ-aminobutyric acid and sialic acid in fermented deer antler velvet and immune promoting effects

  • Yoo, Jiseon;Lee, Juyeon;Zhang, Ming;Mun, Daye;Kang, Minkyoung;Yun, Bohyun;Kim, Yong-An;Kim, Sooah;Oh, Sangnam
    • Journal of Animal Science and Technology
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    • v.64 no.1
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    • pp.166-182
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    • 2022
  • Deer antler velvet is widely used in traditional medicine for its anti-aging, antioxidant, and immunity-enhancing effects. However, few studies have reported on the discovery of probiotic strains for deer antler fermentation to increase functional ingredient absorption. This study evaluated the ability of probiotic lactic acid bacteria to enhance the concentrations of bioactive molecules (e.g., sialic acid and gamma-aminobutyric acid [GABA]) in extracts of deer antler velvet. Seventeen strains of Lactobacillus spp. that were isolated from kimchi and infant feces, including L. sakei, L. rhamnosus, L. brevis, and L. plantarum, and those that improved the life span of Caenorhabditis elegans were selected for evaluation. Of the 17 strains, 2 (L. rhamnosus LFR20-004 and L. sakei LFR20-007) were selected based on data showing that these strains increased both the sialic acid and GABA contents of deer antler extract after fermentation for 2 d and significantly improved the life span of C. elegans. Co-fermentation with both strains further increased the concentrations of sialic acid, GABA, and metabolites such as short-chain fatty acids and amino acids. We evaluated the biological effects of the fermented antler velvet (FAV) on the antibacterial immune response in C. elegans by assessing worm survival after pathogen infection. The survival of the C. elegans conditioned with FAV for 24h was significantly higher compared with that of the control worm group fed only normal feed (non-pathogenic E. coli OP50) exposed to E. coli O157:H7, Salmonella typhi, and Listeria monocytogenes. To evaluate the protective effects of FAV on immune response, cyclophosphamide (Cy), an immune-suppressing agent was treated to in vitro and in vivo. We found that FAV significantly restored viability of mice splenocytes and immune promoting-related cytokines (interleukin [IL]-6, IL-10, inducible nitric oxide synthase [iNOS], interferon [IFN]-γ, and tumor necrosis factor [TNF]-α) were activated compared to non-fermented deer antlers. This finding indicated the protective effect of FAV against Cy-induced cell death and immunosuppressed mice. Taken together, our study suggests that immune-promoting antler velvet can be produced through fermentation using L. rhamnosus LFR20-004 and L. sakei LFR20-007.

An engineered PD-1-based and MMP-2/9-oriented fusion protein exerts potent antitumor effects against melanoma

  • Wei, Mulan;Liu, Xujie;Cao, Chunyu;Yang, Jianlin;Lv, Yafeng;Huang, Jiaojiao;Wang, Yanlin;Qin, Ye
    • BMB Reports
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    • v.51 no.11
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    • pp.572-577
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    • 2018
  • Recent studies showed that the PD-1/PD-L1 checkpoint blockade is a dramatic therapy for melanoma by enhancing antitumor immune activity. Currently, major strategies for the PD-1/PD-L1 blockade have mainly focused on the use of antibodies and compounds. Seeking an alternative approach, others employ endogenous proteins as blocking agents. The extracellular domain of PD-1 (ePD1) includes the binding site with PD-L1. Accordingly, we constructed a PD-1-based recombinantly tailored fusion protein (dFv-ePD1) that consists of bivalent variable fragments (dFv) of an MMP-2/9-targeted antibody and ePD1. The melanoma-binding intensity and antitumor activity were also investigated. We found the intense and selective binding capability of the protein dFv-ePD1 to human melanoma specimens was confirmed by a tissue microarray. In addition, dFv-ePD1 significantly suppressed the migration and invasion of mouse melanoma B16-F1 cells, and displayed cytotoxicity to cancer cells in vitro. Notably, dFv-ePD1 significantly inhibited the growth of mouse melanoma B16-F1 tumor cells in mice and in vivo fluorescence imaging showed that dFv-ePD was gradually accumulated into the B16-F1 tumor. Also the B16-F1 tumor fluorescence intensity at the tumor site was stronger than that of dFv. This study indicates that the recombinant protein dFv-ePD1 has an intensive melanoma-binding capability and exerts potent therapeutic efficacy against melanoma. The novel format of the PD-L1-blocked agent may play an active role in antitumor immunotherapy.

Evaluation of Biological Activities of Fermented Rice Bran from Novel Black Colored Rice Cultivar SuperC3GHi (슈퍼자미(SuperC3GHi) 미강 발효물의 생리활성)

  • Kim, Dong-Ju;Choi, Su-Min;Kim, Hwa-Young;Kim, Joong-Hark;Ryu, Su-Noh;Han, Sang-Jun;Hong, Seong-Gil
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.4
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    • pp.420-426
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    • 2011
  • Rice bran is by-product from the dehulling process of rice, an important staple food in Korea. Various studies have been reported immune-enhancing effects of rice bran cultured with Lentinus edodes. In particular black rice bran that contains anthocyanin has the beneficial effects on immune system. SuperC3GHi, a recently released black colored rice cultivar, has higher contents of cyanidine-3-glucoside (C3G), a strong antioxidant than those of currently cultivated black rice cultivars. In this study, we investigate the beneficial effects of fermented SuperC3GHi rice bran (C3G-F) with Lentinus edodes. Using in vivo and in vitro model for inflammation and allgergy, anti-inflammatory and anti-allergic effects of C3G-F were determined. C3G-F induced tumor suppressive effect on Sarcoma-180 cell and B16 melanoma cell injected to mice. These results suggest that the capacity of C3G-F seems to act as a potent immunomodulator and with the absence of notable side-effects, C3G-F could be used as a novel functional food ingredient having possible therapeutic effects against immunological disorders.