• Title/Summary/Keyword: Linear Cell

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High Concentrated Spore Production of Bacillus thuringliensis by Fed-Batch Processes (유가식 배양공정에 의한 Bacillus thurngiensis의 고농도 포자생산)

  • 박창열;유연우
    • KSBB Journal
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    • v.15 no.3
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    • pp.219-225
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    • 2000
  • Both the production of high spore concentration and high insecticidal activity are required in the production of Bacillus thuringiensis to be used for the bacterial insecticide. In the production of high cell and spore concentrations of B. thuringiensis the continuous fed-batch culture(CFBC) and intermittent fed-batch culture(IFBC) were investigated at $28^{\circ}C$ by maintaining 40% dissolved oxygen concentration. When the final glucose concentration was 50 g/L the maximum viable cell number obtained using the CFBC with linear gradient feeding was $9.37{\times}109$ cells/mL and maximum spore concentration was $8.33{\times}109$ spores/mL which was approximately 84.4% yield of spore formation. When the final glucose concentration was 100 g/L the aximum viable cell and spore concentrations obtained using the IFBC with pH-statb were $1.38{\times}$1010 cells/mL and $1.35{\times}1010$ spores/mL respectively and the yield of spore formation was approximately 97.8%.

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Development of Program for Relative Biological Effectiveness (RBE) Analysis of Particle Beam Therapy

  • Chung, Yoonsun;Ahn, Sang Hee;Choi, Changhoon;Park, Sohee
    • Progress in Medical Physics
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    • v.28 no.1
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    • pp.11-15
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    • 2017
  • Relative biological effectiveness (RBE) of particle beam needs to be evaluated at particle beam therapy centers before the clinical application of the particle beam. However, since RBE analysis is implemented manually, it is useful to have a tool that can easily and effectively handle the data of experiments to generate cell survival curve and to analyze RBE simultaneously. In this work, the development of a program for RBE analysis of particle beam therapy was presented. This RBE analysis program was developed to include two parts; fitting the cell survival curves to linear-quadratic model and calculating the RBE values at a certain endpoint using fitting results. This program was also developed to simultaneously compare and analyze the template results that stored experiment data with photon and particle beam irradiations. The results of the cell survival curve obtained by each irradiation can be analyzed by the user on a desired data after reading the template stored in the easy-to-use excel file. The analysis results include the cell survival curves with error range, which are appeared in the screen and the ${\alpha}$ and ${\beta}$ parameters of linear-quadratic model with 95% confidence intervals, RBE values, and $R^2$ values to evaluate goodness-of-fit of survival curves to model, which are stored in a text cvs file. This software can generate cell survival curve, fit to model, and calculate RBE all at once with raw experiment data, so it helps users to save time for data handling and to reduce the possibility of making error on analysis. As a coming plan, we will create a user-friendly graphical user interface to present the results more intuitively.

Detection of DNA Damage in Carp Using Single-Cell Gel Electrophoresis Assay for Genotoxicity Monitoring

  • Jin, Hai-Hong;Lee, Jae-Hyung;Hyun, Chang-Kee
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.268-275
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    • 2004
  • To investigate the potential application of the single-cell gel electrophoresis (SCGE) assay to carp as an aquatic pollution monitoring technique, gill, liver, and blood cells were isolated from carp exposed to a direct-acting mutagen, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), or indirect mutagen, $benzo[\alpha]pyrene$ $(B[\alpha]P)$, then the DNA strand breakage was analyzed using the assay. Based on testing 5 different cell isolation methods and 6 electrophoretic conditions, the optimized assay conditions were found to be cell isolation by filter pressing and electrophoresis at a lower voltage and longer running time (at 0.4 V/cm for 40 min). In preliminary experiments, gill and liver cells isolated from carp exposed to MNNG in vitro exhibited DNA damage signals even with 0.5 ppb exposure, which is a much higher dose than previously reported. In the gill cells isolated from carp exposed to 0.01-0.5 ppm MNNG in vivo, significant dose-and time-dependent increases were observed in the tail for 4 days. As such, the linear correlation between the relative damage index (RDI) values and time for each dose based on the initial 48-h exposure appeared to provide effective criteria for the genotoxicity monitoring of direct-acting mutagenic pollution. In contrast, the in vivo exposure of carp to 0.25-1.0 ppm of $B[\alpha]P$ for 7 days resulted in dose-and time-dependent responses in the liver cells, in which 24-h delayed responses for metabolizing activation and gradual repair after 48 h were also observed. Thus, the negative-sloped linear correlation between the RDI and time at each dose based on the initial 48 h appeared to provide more effective criteria for the genotoxicity monitoring of indirect mutagenic pollution.

Cellular Uptake and Transfection Efficiency of Plasmid DNA Using Low Molecular Weight Polyethylenimine (저분자량 폴리에틸렌이민을 이용한 유전자 송달 및 발현 유효성 연구)

  • Jeong, Gil-Jae;Park, Kui-Lye;Shin, Ji-Young;Choi, Han-Gon;Oh, Yu-Kyoung
    • Journal of Pharmaceutical Investigation
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    • v.34 no.4
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    • pp.263-267
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    • 2004
  • Branched and linear polyethylenimines (PEIs) have been studied as efficient and versatile agents for gene delivery in vitro and in vivo. PEIs exist in a linear or branched topology and are available in a wide range of molecular weight (Mw). Most studies have been done using PEIs with Mw higher than 10Kd. This study was aimed to test the transfection efficiency and the cell viability following gene delivery using PEI of Mw 2Kd, a relatively lower Mw cationic polymer. We used murine interleukin-2(mIL-2) plasmid DNA complexed with branched PEI 2Kd or 25Kd, and transfected them into a myoblast muscle cell line, C2C12. The cellular uptake of mIL-2 plasmid DNA was determined using quantitative polymerase chain reaction. RNA transcript levels were studied in the myoblast cells. Our results show that PEI 2Kd was as effective as PEI 25Kd in celluar gene delivery and transfection efficiency in C2C12 cells. Moreover, MTT assay indicated that PEI 2Kd/DNA complexes did not significantly reduce the cell viability regardless of N/P ratios. These results suggest that PEI of Mw 2Kd might play a role as effective and low toxic nonviral vector systems for muscular cell lines.

High Cell Density Culture of Anabaena variabilis with Controlled Light Intensity and Nutrient Supply

  • Yoon, Jong-Hyun;Shin, Jong-Hwan;Ahn, Eun-Kyung;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.5
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    • pp.918-925
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    • 2008
  • Controlling the light energy and major nutrients is important for high cell density culture of cyanobacterial cells. The growth phase of Anabaena variabilis can be divided into an exponential growth phase and a deceleration phase. In this study, the cell growth in the deceleration phase showed a linear growth pattern. Both the period of the exponential growth phase and the average cell growth rate in the deceleration phase increased by controlling the light intensity. To control the light intensity, the specific irradiation rate was maintained above $10\;{\mu}mol/s/g$ dry cell by increasing the incident light intensity stepwise. The final cell density increased by controlling the nutrient supply. For the control of the nutrient supply, nitrate, phosphate, and sulfate were intermittently added based on the growth yield, along with the combined control of light intensity and nutrient concentration. Under these control conditions, both final cell concentration and cell productivity increased, to 8.2 g/l and 1.9 g/l/day, respectively.

A Study on Reactions of Palladium Anode Catalyst in Direct Formic Acid Fuel Cells (개미산 연료전지에서 연료극 팔라듐 촉매의 반응에 대한 연구)

  • Han, Jong-Hee;Kim, Jin-Soo;Yoon, Sung-Pil;Nam, Suk-Woo;Lim, Tae-Hoon;Kwon, Yong-Chai
    • Applied Chemistry for Engineering
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    • v.21 no.6
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    • pp.697-701
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    • 2010
  • We investigate the cell performance and characteristics of a direct formic acid fuel cell (DFAFC) using palladium (Pd) as a catalyst for anode. Pd is deposited on the electrolyte using the "direct paint" method. From a continuous three time-test of the polarization curve of the DFAFC, it is found that the catalytic activity of Pd and the cell performance of DFAFC steadily degrade as the tests are proceeded. This behavior may be due to the deactivation of Pd by formate (COOH) and hydroxyl (OH) groups, which are electrochemically dissociated from formic acid solution. Estimations of the degradation, followed by reactivation in activity of Pd catalyst and DFAFC cell performance are implemented by linear voltage sweep tests going in both positive and negative directions. When the maximum voltage of 1.0 V versus DHE is applied to the cell while a linear voltage sweep test going in negative directions, the activity of Pd catalyst and the DFAFC cell performance recover by the rehabilitation in activity of the deactivated Pd.

A Study on Design and Manufacture of an Inchworm Linear Motor System (인치웜 리니어 모터 시스템 설계 및 제작에 관한 연구)

  • Ye Sang Don;Jeong Jae Hoon;Min Byeong Hyeon
    • Journal of the Korean Society for Precision Engineering
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    • v.21 no.12
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    • pp.174-181
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    • 2004
  • Ultra precision positioning mechanism has widely been used on semiconductor manufacturing equipments, optical spectrum analyzer and cell manipulations. Ultra precision positioning mechanism is consisted of several actuators, sensors, guides and control systems. Its efficiency depends on each performance of components. The object of this study is to design, analysis and manufacture all of the inchworm linear motor system, which is one of the equipments embodied in ultra precision positioning mechanism. Inchworm linear motor system is consisted of a controller system and an inchworm linear motor, and its driving form is similar to a motion of spanworm. A design and manufacture of inchworm linear motor, which is consisted of three PZT actuators, a rod, two columns and a guide plate, are performed. Minimizing the von-Mises stress of the hinge using Taguchi method and simulation by FEM software optimizes the structural design in a column of flexure hinge. The designed columns and guide plates are manufactured by a W-EDM and NC-milling. A controller system, which is an apparatus to drive inchworm linear motor, can easily adjust driving conditions by varying resonance frequency and input-output voltage of actuators and amplifiers. The performance of manufactured inchworm linear motor system is verified and valuated. In the future, inchworm linear motor system will be used to make a more precision positioning by reinforcing a sensor and feedback system.

Production of Pyruvate Dehydrogenase Complex-E2 Specific Human Monoclonal Antibody in Fed-batch Culture Systems with High Cell Density Recombinant Escherichia coli (고농도 재조합 대장균의 Fed-batch 배양 시스템을 이용한 Pyruvate Dehydrogenase Complex-E2 특이성 인간 모노클론 항체의 생산)

  • 이미숙;전주미;차상훈;정연호
    • KSBB Journal
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    • v.15 no.5
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    • pp.489-496
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    • 2000
  • Several culture systems including batch, two-stage CSTR, semi-fed batch, and two-stage cyclic fed-batch were investigated for the efficient production of the Fab fraction of PDC-E2 specific human monoclonal antibody using high cell density recombinant E. coli. A two-phase batch system and a two-stage continuous system were examined to overcome plasmid instability problems, by separating the growth and the production stages. The cell density and productivity of the two-stage continuous culture was better than that of the two-phase batch fermentation. In the two-stage continuous culture system with DO-stat, the cell growth and the productivity were superior to those of the system without the DO control. Also, almost total plasmid stability was maintained in the two-stage continuous culture system. Modified M9 medium was selected as an optimum feeding medium for the fed-batch process, and the optimum C/N ratio determined to be 2:3. The optimum feeding rate was $0.6g/\ell/hr$ for a constant feeding strategy in semi-fed batch system. When the feeding medium was fed by pulsing, it was observed that more frequent pulsing resulted in improved cell growth. The linear feeding method was the most efficient of the various feeding methods tested. Finally, high cell density culture using a two-stage cyclic fed batch system with pH-stat was tried because the linear feeding method showed limitations in terms of obtaining high cell densities, and a cell density of $54 g/\ell$ was achieved. It was concluded that the two-stage cyclic fed batch system was the most efficient system for high cell density culture of the systems tested. However, productivity improvements were lower than expected due to the extremely high accumulations of acetate, although the low levels of residual glucose were maintained.

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Non-invasive Methods for Determination of Cellular Growth in Podophyllum hexandrum Suspension Cultures

  • Chattopadhyay, Saurabh;Bisaria, V.S.;Scheper, T.;Srivastava, A.K.
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.6
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    • pp.331-334
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    • 2002
  • Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures of Podophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation In a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the most desirable pH stat conditions.

Efficient Ρ-median approach to GT cell formation (GT 셀 형성을 위한 효율적 Ρ-median 접근법)

  • Won, Youkyung
    • Proceedings of the Korean Operations and Management Science Society Conference
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    • 2000.04a
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    • pp.40-43
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    • 2000
  • This paper is concerned with development of an efficient Ρ-median approach applicable to large cell formation(CF) problems. A two-phase methodology that seeks to minimize the number of exceptional elements is proposed. In phase I, two efficient Ρ-median formulations which contain fewer binary variables than existing Ρ-median formulations are constructed. These make it possible to implement large CF problem within reasonable computer runtime with commercially available linear integer programming codes. Given the initial cell configuration found with the new p-median formulations, in phase II bottleneck machines and parts are reassigned to reduce the number of exceptional elements. This procedure has the flexibility to provide the cell designer with alternative solutions. Test results on large CF problems show a substantial efficiency of the new Ρ-median formulations.

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