• Title/Summary/Keyword: stable cells

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Development of Multiple Beam Optical Tweezers

  • Lee Dong-Jin;LeBrun Thomas W.;Balijepalli Arvind;Gorman JasonJ.;Gagnon Cedric;Hong Dae-Hie;Chang Esthe rH.
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2005.06a
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    • pp.1501-1506
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    • 2005
  • This paper presents the design of a multiple beam optical tweezers instrument used for manipulating micro/nano-sized components. The basic equations used in designing the optical tweezers are derived and the stable and time-sharing multiple beam optical tweezers are constructed with scanning mirrors. The laser beam passes through a series of optical components such as lenses, mirrors, and scanning mirrors, and overfills the entrance aperture of microscope objective, which gives a stable trap. By rotating the laser beam with the scanning mirror, the focal positions are translated in the specimen plane and multiple micro/nano-sized objects can be moved. The constructed optical tweezers is used to manipulate cells and liposomes simultaneously and to trap multiple nano-wires. The experiments prove that the developed optical tweezers can be a very versatile manipulation tool for studying gene therapy and nano device fabrication.

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A Physics-Based Modelling of Multipbase Fluid Phenomena (물리적 모델에 기반한 다상 유체 현상 애니메이션)

  • Song, Oh-Young;Shin, Hyun-Cheol;Ko, Hyeong-Seok
    • Journal of the Korea Computer Graphics Society
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    • v.10 no.3
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    • pp.52-60
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    • 2004
  • This paper presents a physically based technique for simulating complex multiphase fluids. This work is motivated by the "stable fluids" method developed by Stam to handle gaseous fluids. We extend this technique to water, which calls for the development of methods for modeling multiphase fluids and suppressing dissipation. We construct a multiphase fluid formulation by combining the Navier-Stokes equations with the level set method. By adopting constrained interpolation profile (CIP)-based advection, we reduce the numerical dissipation and diffusion significantly. We further reduce the dissipation by converting potentially dissipative cells into droplets or bubbles that undergo Lagrangian motion. Due to the multiphase formulation, the proposed method properly simulates the interaction of water with surrounding air, instead of simulating water in a void space. Moreover. the introduction of the non-dissipative technique means that, in contrast 10 previous methods, the simulated water does not unnecessarily lose mass and its motion is not damped to an unphysical extent. Experiments showed that the proposed method is stable and runs fast. It is demonstrated that two-dimensional simulation runs in real-time.

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Characterization of Proteus vulgaris K80 Lipase Immobilized on Amine-Terminated Magnetic Microparticles

  • Natalia, Agnes;Kristiani, Lidya;Kim, Hyung Kwoun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.10
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    • pp.1382-1388
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    • 2014
  • Proteus vulgaris K80 lipase was expressed in Escherichia coli BL21 (DE3) cells and immobilized on amine-terminated magnetic microparticles (Mag-MPs). The immobilization yield and activity retention were 84.15% and 7.87%, respectively. A homology model of lipase K80 was constructed using P. mirabilis lipase as the template. Many lysine residues were located on the protein surface, remote from active sites. The biochemical characteristics of immobilized lipase K80 were compared with the soluble free form of lipase K80. The optimum temperature of K80-Mag-MPs was $60^{\circ}C$, which was $20^{\circ}C$ higher than that of the soluble form. K80-Mag-MPs also tended to be more stable than the soluble form at elevated temperatures and a broad range of pH. K80-Mag-MP maintained its stable form at up to $40^{\circ}C$ and in a pH range of 5.0-10.0, whereas soluble K80 maintained its activity up to $35^{\circ}C$ and pH 6.0-10.0. K80-Mag-MPs had broader substrate specificity compared with that of soluble K80. K80-Mag-MPs showed about 80% residual relative activity after five recovery trials. These results indicate the potential benefit of K80-Mag-MPs as a biocatalyst in various industries.

Design of a control scheme for applying DC power sources to a distribution system (배전시스템에 DC 전력원을 적용하기 위한 제어 기법 설계)

  • Hwang, Chul-Sang;Kim, Gyeong-Hun;Byeon, Gilsung;Jeon, Jin-Hong;Jo, Chang-Hee;Park, Minwon;Yu, In-Keun
    • Proceedings of the KIEE Conference
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    • 2015.07a
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    • pp.1056-1057
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    • 2015
  • A common DC bus is a useful connection for several DC output sources such as photovoltaic (PV), fuel cells, and batteries. Operation of the common DC power system with more than two DC output sources, especially in a stand-alone mode, requires a control scheme for the stable operation of the system. In this paper, a control scheme has been developed for applying DC power sources to the distribution system. The purpose of the control scheme is to make the best use of the DC power sources. The DC power system consists of PV, two energy storage systems and a DC-AC inverter with the control scheme. A distribution system was modeled in PSCAD/EMTDC. As the results, the control scheme is applied to the DC-AC inverter and the DC-DC converter for transfer operations between the grid-connected and the stand-alone mode to keep the DC bus and the AC voltage constant. The results from the simulation demonstrate the stable operation of a grid connected DC power system.

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Lentivirus Mediated GOLPH3 shRNA Inhibits Growth and Metastasis of Esophageal Squamous Cancer

  • Wang, Qiang;Wang, Xian;Zhang, Can-Bin
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5391-5396
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    • 2013
  • Aim: To investigate the role of Golgi phosphoprotein 3 (GOLPH3) in tumour growth and metastasis of esophageal squamous cancer. Methods: A lentiviral shRNA-vector was utilized to stably knockdown GOLPH3 in Eca-109 esophageal squamous cancer cells. mRNA transcription and protein expression of GOLPH3 were examined by real-time quantitative PCR and Western blotting, respectively. Cell proliferation activity was assessed by MTT assay and invasion and migration potentials by matrigel invasion and transwell motility assays. Results: Stable knockdown in the GOLPH3 cell line was established. PD-A gene expression was significantly suppressed by lentivirus-mediated RNAi, which resulted in reducing the capacity for cell proliferation, migration, invasion and adhesion in vitro. In vivo, GOLPH3 depletion resulted in inhibition of tumour growth, with stable decrease in the expression of GOLPH3 in tumor xenografts. Conclusions: Our findings suggest that lentivirus mediated silencing of the GOLPH3 gene has a significant anti-tumour effect on esophageal squamous cancer in vitro and in vivo. In addition, the results indicate that GOLPH3 might be an effective molecular target for gene therapy in esophageal squamous cancer.

Deciphering Functions of Uncultured Microorganisms (난배양성 미생물의 기능 분석 방법)

  • Kim, Jeong-Myeong;Song, Sae-Mi;Jeon, Che-Ok
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.1-9
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    • 2009
  • Microbes within complex communities show quite different physiology from pure cultured microbes. However, historically the study of microbes has focused on single species in pure culture and most of microbes are unculturable in our labs, so understanding of complex communities lags behind understanding of pure cultured cells. Methodologies including stable isotope probing (SIP), a combination of fluorescence in situ hybridization (FISH) and microautoradiography (MAR), isotope micrarray, and metagenomics have given insights into the uncultivated majority to link phylogenetic and functional information. Here, we review some of the most recent literatures, with an emphasis on methodological improvements to the sensitivity and utilities of these methods to link phylogeny and function in complex microbial communities.

Anti-Cariogenicity of NCS (Non-Cariogenicity Sugar) Produced by Alkalophilic Bacillus sp. S-1013

  • Ryu, Il-Hwan;Kim, Sun-Sook;Lee, Kap-Sang
    • Journal of Microbiology and Biotechnology
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    • v.14 no.4
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    • pp.759-765
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    • 2004
  • The NCS inhibited the activity of glucosyltransferase which was produced by Streptococcus mutans JC-2, and the rate of inhibition at $100\muM<$ and $200\muM$ were 74.0% and 99.8%, respectively. It was stable in alkali condition, but unstable in acid condition. It was also stable up to $80^{\circ}C$. The kinetic study of the inhibition by NCS was carried out by Lineweaver-Burk plot and Dixon plot. It was non-competitive inhibition, determined by the two plots and $K_i$ and $K_i$ values were $15\muM$ and $19.3\muM$ respectively. The NCS did not show cytotoxicity against human gingival cells at $K_i$ ($15\muM$, $150\muM$, $1,500\mu$ M) concentrations. It had less cytotoxicity than chlohexidin, which has usually been used as the agent of anticaries. To evaluate the industrial applicability of the NCS, human pluck tooth was used. The inhibitory rates of tooth calcification and calcium ion elution by the NCS were 41 % and 2.5 times, respectively. These results suggested that NCS from Bacillus sp. S-1013 is an efficient anticaries agent.

Characterization of a Novel Carbohydrase from Lipomyces starkeyi KSM 22 for Dental Application

  • KIM, DOMAN;SU-JIN RYU;SOO-JIN HEO;DO-WON KIM;HO-SANG KIM
    • Journal of Microbiology and Biotechnology
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    • v.9 no.3
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    • pp.260-264
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    • 1999
  • The combined activities of dextranase and amylase(DXAMase) from Lipomyces starkeyi KSM 22 produced from starch fermentation inhibited or prevented dental plaque formation. The activities were stable in commercial mouthwash products; DXAMase activity retained over 93% of original activity after 6 months at 23℃. We examined the effects of enzyme inhibitors and active ingredients in mouthwash on DXAMase activity. The DXAMase was stable with 0.29%(w/v) EDTA, 20% (v/v) ethanol, 0.05% (w/v) fluoride, and 0.05% (w/v) SDS. Among the active ingredients of mouthwash, sodium benzoate (up to 1 %, w/v) had no inhibitory effect on either dextranase or amylase activity. In the case of cetylpyridinium chloride, the addition of 0.05% (w/v) inhibited 6% of dextranase activity and 13% of amylase activity. Propylene glycol (up to 1%, w/v) showed no inhibitory effect on either enzyme activity. DXAMase (5 IU/㎖) in mouthwash could remove pre-formed films of glucan-bound S. mutans cells. The addition of 0.1 IU/㎖ DXAMase in mouthwash prevented the formation of insoluble-glucan. These in vitro properties of L. starkeyi KSM 22 DXAMase are desirable for its application as a dental plaque control agent.

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New Cationic Liposome with Enhanced Stability and Transfection Efficiency for Gene Delivery (안정성 및 Transfection 효율이 우수한 양이온성 리포좀 유전자 전달시스템의 개발)

  • Kim, Kyoung-Mi;Nam, Bang-Hyun;Sohn, Dong-Hwan
    • Journal of Pharmaceutical Investigation
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    • v.28 no.2
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    • pp.93-98
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    • 1998
  • We have developed liposomes which can be easily prepared with inexpensive lipid, have enhanced stability, and can efficiently deliver DNA into the COS-l cells, Liposome formulations were prepared using cationic materials such as dimethyldioctadecyl ammonium bromide (DDAB), cetyltrimethyl ammonium bromide(CTAB), We investigated the effect of cationic liposome formulations on in vitro DNA transfection, DDAB-containing liposomes showed increased transfection efficiency which was 3.2-fold as much as that by $Lipofectin^{\circledR}$, but CTAB-containing liposomes were inactive in gene transfection. The effect of colipid of DDAB-containing liposome was also investegated. As a colipid, dioleylphosphatidylethanolamine(DOPE) and cholesterol did altered the transfection efficiency of DDAB-containing liposomes. And increased DDAB concentration lowered the transfection efficiency. The optimum amount of liposomal formulation was $10\;{\mu}M$ for $1\;{\mu}g$ of DNA. In the experiment of stability, DOPE-containing liposomes formulation showed a broad size distribution and separation of two major peaks on a 5th day of preparation, but liposomes containing cholesterol was stable for 10 days. DDAB-containing liposomal DNA delivery system was prepared easily and was stable.

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Transformation of a Filamentous Fungus Cryphonectria parasitica Using Agrobacterium tumefaciens

  • Park, Seung-Moon;Kim, Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.9 no.3
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    • pp.217-222
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    • 2004
  • As Agrobacterium tumefaciens, which has long been used to transform plants, is known to transfer T-DNA to budding yeast, Saccharomyces cerevisiae, a variety of fungi were subjected to the A. tumefaciens-mediated transformation to improve their transformation frequency and feasibility. The A. tumefaciens-mediated transformation of chestnut blight fungus, Cryphonectria parasitica, is performed in this study as the first example of transformation of a hardwood fungal pathogen. The transfer of the binary vector pBIN9-Hg, containing the bacterial hygromycin B phosphotransferase gene under the control of the Aspergillus nidulans trpC promoter and terminator, as a selectable marker, led to the selection of more than 1,000 stable, hygromycin B-resistant transformants per 1${\times}$10$\^$6/ conidia of C. parasitica. The putative transformants appeared to be mitotically stable. The transformation efficiency appears to depend on the bacterial strain, age of the bacteria cell culture and ratio of fungal spores to bacterial cells. PCR and Southern blot analysis indicated that the marker gene was inserted at different chromosomal sites. Moreover, three transformants out of ten showed more than two hybridizing bands, suggesting more than two copies of the inserted marker gene are not uncommon.