• Title/Summary/Keyword: Medium Barrier

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Enhanced Production of hCTLA4Ig through Increased Permeability in Transgenic Rice Cell Cultures (형질전환 벼 현탁세포 배양에서 투과성 증진을 통한 hCTLA4Ig의 생산성 증대)

  • Choi, Hong-Yeol;Cheon, Su-Hwan;Kwon, Jun-Young;Lim, Jung-Ae;Park, Hye-Rim;Kim, Dong-Il
    • KSBB Journal
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    • v.31 no.4
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    • pp.277-283
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    • 2016
  • In this system, rice cells were genetically modified to express human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) using RAmy3D promoter induced by sugar depletion. Even though the target protein fused with signal sequence peptide, plant cell wall can be a barrier against secretion of recombinant proteins. Therefore, hCTLA4Ig can be trapped inside cell wall or remained in intracellular space. In this study, to enhance the secretion of hCTLA4Ig from cytoplasm and cell walls into the medium, permeabilizing agents, such as dimethyl sulfoxide (DMSO), Triton X-100 and Tween 20, were applied in transgenic rice cell cultures. When 0.5% (v/v) of DMSO was added in sugar-free medium, intracellullar hCTLA4Ig was increased, on the other hand, the secreted extracellular hCTLA4Ig was lower than that of control. DMSO did not give permeable effects on transgenic rice cell cultures. And Triton X-100 was toxic to rice cells and also did not give enhancing permeability of cells. When 0.05% (v/v) Tween 20 was added in rice cell cultures, however, intracellular hCTLA4Ig was lower than that of control cultures. And the maximum 44.76 mg/L hCTLA4Ig was produced for 10 days after induction, which was 1.4-fold increase compared to that of control cultures. Especially, Tween 20 at 0.05% (v/v) showed the positive effect on the secretion of hCTLA4Ig though the decrease of intracellular hCTLA4Ig. Also, Tween 20 as a non-toxic surfactant did not affect the cell growth, cell viability and protease activity. In conclusion, secretion of hCTLA4Ig could be increased by enhancing permeability of cells regardless of the cell growth, cell viability and protease activity.

Enhanced Skin Permeation of a New Capsaicin Derivative (DA-5018) from a Binary Vehicle System Composed of Isopropyl-myristate and Ethoxydiglycol

  • Cha, Bong-Jin;Lee, Eung-Doo;Kim, Won-Bae;Chung, Suk-Jae;Lee, Min-Hwa;Shim, Chang-Koo
    • Archives of Pharmacal Research
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    • v.24 no.3
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    • pp.224-228
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    • 2001
  • DA-5018, a recently synthesized capsaicin analog, appears to possess potent analgesic activity when administered topically. The objective of this study is to test the feasibility of the topical administration of this compound. Specifically, our goal was to identify vehicle system that permit a reasonable transdermal permeation of the compound in mice. Among the vehicles examined, isopropyl myristate (IPM) showed the largest in vitro permeability across the intact skin (83.6 ${\pm}$ 5.42${\mu}$l/$\textrm{cm}^2$/h ). However, due to the limited solubility of DA-5018 in IPM (0.53 mg/ml), the maximal flux from the IPM medium remained at only 44.3 ${\pm}2.87{\mu}$g/$\textrm{cm}^2$/hr. In order to increase the flux, addition of better solvents for DA-5018 was attempted, under the assumption that flux is the result of both solubility and permeability. Ethoxydiglycol (EG) and oleic acid (OA) were selected as examples of food solvents. The addition of IC or OA to IPM at a 1:1 volume ratio resulted in a comparable increase in the solubility of the compound (i.e., to 61.1 and 50.2 mg/ml for EG and OA, respectively). However, the addition of EG at a 1:1 volume ratio, for example, increased the flux 6.3 fold (i.e., $279{\mu}$g/$\textrm{cm}^2$/hr), while OA, at a 1:1 volume ratio, decreased the flux 5 fold (i.e., $9.26{\mu}$g/$\textrm{cm}^2$//hr). The mechanism of this discrepancy between EG and OA was investigated by measuring the permeabilty of DA-5018 across the stratum corneum-removed skin of the mouse, under the hypothesis that the viable skin layer may serve as a barrier for the permeation of lipophilic substances such as DA 5018. The permeability of DA-5018, from the medium of EG or OA, across the viable skin differed greatly for EG ($0.41{\mu}$l/$\textrm{cm}^2$/hr) and OA ($0.086{\mu}$l/$\textrm{cm}^2$/hr), suggesting that a higher permeability across the viable skin layer is needed for the second solvents. The maximum flux across the intact skin was achieved for DA-5018 when EG was added to IPM at a 1:1 volume ratio. Thus, the use of a binary system appears to be the best approach for realizing the transdermal delivery of DA-5018 at a reasonable rate.

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Hair Revitalization Effects of Streptococcus sp. Strain Culture Medium Filtrate (HAIRCLETM) (Streptococcus sp. 배양 여과물(HAIRCLETM)의 모발 개선 효과)

  • Park, Hye Rim;Kim, Ha Yeon;Kim, Jin Young;Oh, Sinseok;Kang, Pilsun;Lee, Gang Hyuk;Lim, Jung-Hyurk;Shin, Song Seok
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.48 no.2
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    • pp.169-180
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    • 2022
  • Streptococcus sp. is generally known as a strain that produces hyaluronic acid. In this study, we focused on ingredients other than hyaluronic acid among several metabolites produced by Streptococcus sp. during cultivation. The hair experimental sample (HAIRCLETM) was prepared with Streptococcus sp. culture filtrate under the condition that hyaluronic acid was not produced and the effect on the hair was identified. Tensile strength of hair was tested with a universal testing machine (UTM). Surface properties of a hair were examined with an atomic force microscope (AFM). Our results suggest that HAIRCLETM could prevent damage to a hair such as destroyed-cuticle, increased-faction force, and broken-hair. Furthermore, hair-related experiments confirmed that HAIRCLETM could promote the expression of hair growth factors VEGF, HGF, and Wnt10b and mitigate male hair loss by inhibiting androgen receptors and TGF-β2 expression. In addition, it was confirmed that the expression of barrier-related factors (INV, CLDN-1) was increased for oxidative stress, and the COX-2 expression, an inflammatory factor, was inhibited, thereby improving damaged scalp barriers and alleviating induced inflammation.

Rutin alleviated lipopolysaccharide-induced damage in goat rumen epithelial cells

  • Jinshun Zhan;Zhiyong Gu;Haibo Wang;Yuhang Liu;Yanping Wu;Junhong Huo
    • Animal Bioscience
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    • v.37 no.2
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    • pp.303-314
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    • 2024
  • Objective: Rutin, also called vitamin P, is a flavonoids from plants. Previous studies have indicated that rutin can alleviate the injury of tissues and cells by inhibiting oxidative stress and ameliorating inflammation. There is no report on the protective effects of rutin on goat rumen epithelial cells (GRECs) at present. Hence, we investigated whether rutin can alleviate lipopolysaccharide (LPS)-induced damage in GRECs. Methods: GRECs were cultured in basal medium or basal medium containing 1 ㎍/mL LPS, or 1 ㎍/mL LPS and 20 ㎍/mL rutin. Six replicates were performed for each group. After 3-h culture, the GRECs were harvested to detect the relevant parameters. Results: Rutin significantly enhanced the cell activity (p<0.05) and transepithelial electrical resistance (TEER) (p<0.01) and significantly reduced the apoptosis rate (p<0.05) of LPS-induced GRECs. Rutin significantly increased superoxide dismutase, glutathione peroxidase, and catalase activity (p<0.01) and significantly decreased lactate dehydrogenase activity and reactive oxygen species and malondialdehyde (MDA) levels in LPS-induced GRECs (p<0.01). The mRNA and protein levels of interleukin 6 (IL-6), IL-1β, and C-X-C motif chemokine ligand 8 (CXCL8) and the mRNA level of tumor necrosis factor-α (TNF-α) and chemokine C-C motif ligand 5 (CCL5) were significantly increased in LPS-induced GRECs (p<0.05 or p<0.01), while rutin supplementation significantly decreased the mRNA and protein levels of IL-6, TNF-α, and CXCL8 in LPS-induced GRECs (p<0.05 or p<0.01). The mRNA level of toll-like receptor 2 (TLR2), and the mRNA and protein levels of TLR4 and nuclear factor κB (NF-κB) was significantly improved in LPS-induced GRECs (p<0.05 or p<0.01), whereas rutin supplementation could significantly reduce the mRNA and protein levels of TLR4 (p<0.05 or p<0.01). In addition, rutin had a tendency of decreasing the protein levels of CXCL6, NF-κB, and inhibitor of nuclear factor kappa-B alpha (0.05

Reduction of RDX in Ground Water by Bio-Regenerated Iron Mineral: Results of Field Verification Test at a Miliary Shooting Range (생물환원 철광물촉매에 의한 지하수 내 RDX 환원:군사격장 현장적용 실증결과)

  • Gong, Hyo-young;Lee, Kwang-pyo;Lee, Jong-yeol;Kyung, Daeseung;Lee, Woojin;Bae, Bumhan
    • Journal of Soil and Groundwater Environment
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    • v.20 no.6
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    • pp.62-72
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    • 2015
  • This study investigates the in-situ implementation of bio-regenerated iron mineral catalyst to remove explosive compounds in ground water at a military shooting range in operation. A bio-regenerated iron mineral catalyst was synthesized using lepidocrocite (iron-bearing soil mineral), iron-reducing bacteria Shewanella putrefaciens CN32, and electron mediator (riboflavin) in the culture medium. This catalyst was then injected periodically in the ground to build a redox active zone acting like permeable reactive barrier through injection wells constructed at a live fire military shooting range. Ground water and core soils were sampled periodically for analysis of explosive compounds, mainly RDX and its metabolites, along with toxicity analysis and REDOX potential measurement. Results suggested that a redox active zone was formed in the subsurface in which contaminated ground water flows through. Concentration of RDX as well as toxicity (% inhibition) of ground water decreased in the downstream compared to those in the upstream while concentration of RDX reduction products increased in the downstream.

Post Closure Long Term Safely of the Initial Container Failure Scenario for a Potential HLW Repository (고준위 방사성폐기물 처분장 불량 용기 발생 시나리오에 대한 폐쇄후 장기 방사선적 안전성 평가)

  • 황용수;서은진;이연명;강철형
    • Journal of Nuclear Fuel Cycle and Waste Technology(JNFCWT)
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    • v.2 no.2
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    • pp.105-112
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    • 2004
  • A waste container, one of the key components of a multi-barrier system in a potential high level radioactive waste (HLW) repository in Korea ensures the mechanical stability against the lithostatic pressure of a deep geologic medium and the swelling pressure of the bentonite buffer. Also, it delays potential release of radionuclides for a certain period of time, before it is corroded by intruding impurities. Even though the material of a waste container is carefully chosen and its manufacturing processes are under quality assurance processes, there is a possibility of initial defects in a waste container during manufacturing. Also, during the deposition of a waste container in a repository, there is a chance of an incident affecting the integrity of a waste container. In this study, the appropriate Features, Events, and Processes(FEP's) to describe these incidents and the associated scenario on radionuclide release from a container to the biosphere are developed. Then the total system performance assessment on the Initial waste Container Failure (ICF) scenario was carried out by the MASCOT-K, one of the probabilistic safety assessment tools KAERI has developed. Results show that for the data set used in this paper, the annual individual dose for the ICF scenario meets the Korean regulation on the post closure radiological safety of a repository.

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Plant responses to nano and micro structured carbon allotropes: Water imbibition by maize seeds upon exposure to multiwalled carbon nanotubes and activated carbon

  • Dasgupta-Schubert, N.;Tiwari, D.K.;Francis, E. Reyes;Martinez Torres, P.;Villasenor Cendejas, L.M.;Lara Romero, J.;Villasenor Mora, C.
    • Advances in nano research
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    • v.5 no.3
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    • pp.245-251
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    • 2017
  • Multiwalled carbon-nanotubes (MWCNT) and micro-structured carbon, such as biochar or activated carbon (AC), have been seen to significantly increase the growth indices of certain plant species such as maize (Zea mays L.). Seed imbibition is the stage where environmental factors that affect water transport across the seed coat barrier, make a large impact. This work explores the effect on water imbibition by maize seeds when the aqueous environment surrounding the seed is diluted by small concentrations (10 and 20 mg/l) of pristine MWCNT (p-MWCNT), carboxylate functionalized MWCNT (COO-MWCNT) and AC. The degree of sensitivity of the process to (i) large structural changes is seen by utilizing the nano (the MWCNT) and the micro (the AC) allotropic forms of carbon; (ii) to small changes in the purity and morphology of the p-MWCNT by utilizing 95% pure and 99% pure p-MWCNTs of slightly differing morphologies; and (iii) to MWCNT functionalization by using highly pure (97%) COO-MWCNT. Water imbibition was monitored over a 15 hour period by Near Infrared Thermography (NIRT) and also by seed weighing. Seed surface topography was seen by SEM imaging. Analysis of the NIRT images suggests rapid seed surface topological changes with the quantity of water imbibed. While further work is necessary to arrive at a conclusive answer, this work shows that the imbibition phase of the maize seed is sensitive to the presence of MWCNT even to small differences in the purity of the p-MWCNT and to small differences in the physicochemical properties of the medium caused by the hydrophilic COO-MWCNT.

A Release Test for Gels Containing Ketoprofen (겔제제로부터 케토프로펜의 방출특성 시험법)

  • Kim, Ho-Jeong;Yun, Mi-Ok;Lee, Su-Jung;Choi, Hyun-Chul;Kim, Ji-Young;Kim, In-Wha;Shim, Chang-Koo;Kang, Shin-Jung
    • Journal of Pharmaceutical Investigation
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    • v.32 no.2
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    • pp.103-106
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    • 2002
  • A method that describes the determination of the in vitro release of ketoprofen from gels was suggested. The experimental system of the method consists of a Franz diffusion cell, which contains a pH 7.4 phosphate buffer as a receptor medium, and a $70\;{\mu}m$ mesh woven nylon membrane as a diffusion barrier. Under the given condition of the system, the diffusion of ketoprofen across the membrane was rapid enough that the apparent release profile of ketoprofen obtained from the present method could represent the release of the drug from gel preparations. The release of ketoprofen in the present method was reproducible, and the rate increased in proportion to the concentration of ketoprofen in the gel. These suggest that the present method is applicable to the quality evaluation of gel preparations containing ketoprofen.

Wired/Wireless Gateway System Supporting LAN-to-LAN VPN with Multi-Queuing Realtime Traffic Shaping (다중큐잉 실시간 트래픽쉐이핑을 적용한 네트워크간 VPN 지원 유무선공유기 시스템)

  • Yang, Seung Eui;Goh, Byung Oh;Choi, Jong-Kun;Jung, Hoe-kyung
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.19 no.5
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    • pp.1097-1103
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    • 2015
  • In order to build network infrastructure to implement the aforementioned advantages enabling smart device users to work anywhere, professional support and expensive VPN devices are required. This is a barrier to supplying VPN devices to small and medium-sized institutes. To address this issue, this study aims to implement OpenVPN, OpenSSH and iproute based on the OpenWRT platform which is an embedded OS for open networks in affordable open wired/wireless gateway H/W platforms to support the inter-network VPN. In addition, the network environment can be maintained optimal by applying a "multi-queuing real-time traffic shaping technology" to VPN tunnels, although channel quality changes.

Effect of Expression of Genes in the Sphingolipid Synthesis Pathway on the Biosynthesis of Ceramide in Saccharomyces cerevisiae

  • Kim, Se-Kyung;Noh, Yong-Ho;Koo, Ja-Ryong;Yun, Hyun-Shik
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.356-362
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    • 2010
  • Ceramide is important not only for the maintenance of the barrier function of the skin but also for the water-binding capacity of the stratum corneum. Although the exact role of ceramide in the human skin is not fully understood, ceramide has become a widely used ingredient in cosmetic and pharmaceutical industries. Compared with other microorganisms, yeast is more suitable for the production of ceramide because yeast grows fast and is non-toxic. However, production of ceramide from yeast has not been widely studied and most work in this area has been carried out using Saccharomyces cerevisiae. Regulating the genes that are involved in sphingolipid synthesis is necessary to increase ceramide production. In this study, we investigated the effect of the genes involved in the synthesis of ceramide, lcb1, lcb2, tsc10, lac1, lag1, and sur2, on ceramide production levels. The genes were cloned into pYES2 high copy number vectors. S. cerevisiae was cultivated on YPDG medium at $30^{\circ}C$. Ceramide was purified from the cell extracts by solvent extraction and the ceramide content was analyzed by HPLC using ELSD. The maximum production of ceramide (9.8 mg ceramide/g cell) was obtained when the tsc10 gene was amplified by the pYES2 vector. Real-time RT-PCR analysis showed that the increase in ceramide content was proportional to the increase in the tsc10 gene expression level, which was 4.56 times higher than that of the control strain.