• Title/Summary/Keyword: maximal condition

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Characterization of the nar Promoter of Escherichia coli to use as an inducible promoter in Wild-type host Agrobacterium.tumefaciens

  • Lee, Gil-Ho;Jo, Mu-Hwan;Lee, Jong-Won
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.758-761
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    • 2001
  • In this study, the nar promoter of E. coli was characterized to see whether the nar promoter cloned onto pBBR122 can be used as an expression promoter of gram negative microbes. For this purpose, a plasmid with lacZ gene expressing ${\beta}-galactosidase$ instead of the structural genes of nar operon in a gram negative host strain(Agrobacterium.tumefaciens) was used to simplify an assay of induction of the nar promoter. The following effects were investigated to find optimal conditions: methods of inducing the nar promoter, optimal nitrate concentration, maximally inducing the nar promoter, the amount of expressed ${\beta}-galactosidase$ and induction ratio(specific ${\beta}-galactosidase$ activity after maximal induction/specific ${\beta}-galactosidase$ activity before induction). The following results were obtained from the experiments: the growth of Agrobacterium with E.coli nar promoter was not much affected by nitrate concentration in the shake-flask; induction of nar promoter was optimal when Agrobacterium was grown in the presence of 1% nitrate ion at the beginning of culture and when overnight culture was completely grown in the shake-flask before being transferred to other shake-flask; the amount of ${\beta}-galactosidase$ per cell and per medium volume was maximal when Agrobacterium was grown under aerobic condition to $OD_{600}$ of 1.7; then the nar promoter was induced under microaerobic and anaerobic condition made by lowering dissolved oxygen level(DO). After 2-3h of induction in the YEP medium selected as a main culture medium, the specific ${\beta}-galactosidase$ activity became about 17,000 Miller units in the fermentor cluture.

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Production of poly-$\beta$-hydroxybutyric acid(PHB) from Liquefied Natural Gas using an Obligatory Methanotroph Methylosinus trichosporium OB3b (메탄자화균 Methylosinus trichosporium OB3b를 이용한 액화 천연가스로부터 poly-$\beta$-hydroxybutyric acid(PHB)의 생산)

  • 황재웅;박성훈
    • KSBB Journal
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    • v.11 no.2
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    • pp.246-253
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    • 1996
  • An obligatory methanotroph Methylosinus trichosporium OB3b was cultivated for the production of poly-${\beta}$-hydroxybutyric acid(PHB) in shake-flask using liquefied natural gas(LNG) as the sole source of carbon and energy. The maximal specific growth rate decreased by 40% using LNG compared with that obtained with pure methane. This is attributed to the inhibition by ethane and propane presents in the LNG as impurities. For the production of PHB, two-stage culture separating the production stage from the growth stage was carried out. PHB accumulation was observed after switching nutrient-sufficient to nutrient-limited condition of non-carboneous component (NO3-, PO43-, K+, Na+, Fe2+, or Mg2+). The limitation of K+ or Mg2+ resulted in relatively high PHB content, but the highest content was obtained by nitrate limitation. The optimal pH and temperature for PHB accumulation was 7.0 and $30^{\circ}C$. Under the optimal condition the maximal PHB content was about 45% after 4-day cultivation.

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Normal Walking Versus Toe-walking in Healthy Subjects: An Electromyographic Analysis (정상 보행과 발가락 보행의 하지 근육 근 활성도 비교)

  • Kim, Tack-Hoon;Choi, Houng-Sik;Kim, Chang-In;Yi, Jin-Bock
    • Physical Therapy Korea
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    • v.9 no.2
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    • pp.43-50
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    • 2002
  • This study was designed to identify the effects of walking conditions (normal walking vs. toe-walking) on electromyographic (EMG) activity of gastrocnemius, tibialis anterior, and soleus muscle. Seven healthy adult males participated in this study. The exclusion criteria were orthopedic or neurologic disease, congenital anomaly or acquired deformity, or pain in low back or lower extremities. The maximal voluntary isometric contraction for each muscle was used for the reference contraction, and EMG activity of each muscle during normal walking and toe-walking was expressed as a percentage of reference contraction. The gait cycle was determined with two foot switches, and gait was normalized as 100% gait cycle for each condition. The maximal values of EMG activity in terminal stance (30~50% of gait cycle) of each condition were compared for data analysis. No significant differences were found in EMG activity of the tibialis anterior and soleus (p>.05), whereas significant decrement was found in EMG activity of gastrocnemius during toe-walking compared to normal walking (p<.05). There is a limitation to generalize the results of this study, because small number of subjects participated for this study and only EMG was used for data collection. The treatment methods should be developed to improve gait efficiency by substituting the weakened muscles secondary to upper motor neuron, or by strengthening the distal muscles in lower extremity.

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Effect of Muscle Pre-activation Properties on the Magnitude of Joint Torque during Voluntary Isometric Knee Extension (등척성 무릎 토크 발생 시 사전활성화 유형의 차이가 최대 자발적 토크 생성에 미치는 영향)

  • Kim, Jong-Ah;Shin, Narae;Lee, Sungjune;Xu, Dayuan;Park, Jaebum
    • Korean Journal of Applied Biomechanics
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    • v.31 no.2
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    • pp.140-147
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    • 2021
  • Objective: The purpose of this study is to identify the mechanism of changes in maximum voluntary torque with the magnitude and duration of pre-activation torque during voluntary isometric knee extension. Method: 11 male subjects (age: 25.91±2.43 yrs., height: 173.12±3.51 cm, weight: 76.45±7.74 kg) participated in this study. The subjects were required to produce maximal voluntary isometric torque with a particular pre-activation torque condition. The properties of pre-activation torque consisted of the combinations of 1) three levels of magnitude, e.g., 32 Nm, 64 Nm, 96 Nm, and 2) two levels of duration, e.g., 1 sec, and 3 sec; thus, a total of six conditions were given to the subjects. The force and EMG data were measured using the force transducers and wireless EMG sensor, respectively. Results: The results showed that the maximum voluntary torque increased the most with relatively large and fast (96 Nm, 1 sec) pre-activation condition. Similarly, with relatively large and fast (96 Nm, 1 sec) preactivation, it was found that the integrated EMG (iEMG) of the agonist muscles increased, while no significant changes in the co-contraction of the antagonist muscles for the knee extension. Also, the effect of pre-activation conditions on the rate of torque development was not statistically significant. Conclusion: The current findings suggest that relatively larger in magnitude and shorter in duration as the properties of pre-activation lead to a larger magnitude of maximal voluntary torque, possibly due to the increased activity of the agonist muscles during knee extension.

Plasmid Stability and Cloned-Gene Expression in Continuous Culture of Recombinant Escherichia Coli Under Derepressed Condition

  • Nam, Soo-Wan;Kim, Byung-Kwan;Kim, Jung-Hoe
    • Journal of Microbiology and Biotechnology
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    • v.4 no.1
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    • pp.1-6
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    • 1994
  • Continuous culture was carried out with a recombinant Escherichia coli W3110/pCR185, which encodes trp-operon enzymes when the temperature is shifted from $37^{circ}C\;t;42^{\circ}C$. Under derepressed condition of $42^{\circ}C$. plasmlid stability and gene expression were analysed as function of the dilution rate. The stability of plasmid increased with the dilution rate, but maximal levels of gene expression (tryptophan concentration) and plasmid DNA content were obtained at the lowest dilution rate, $0.075\;hr^{-1}$. The plasmid instability, observed at low dilution rates, could be explained by the unbalanced biosynthetic state of the recombinant cell harboring a high copy number of plasmid.

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Effects of Environmental Factors on the Zoospore Release and Germling Growth of the Green Macroalga Chaetomorpha linum (녹조류 실염주말의 유주자 방출과 배아생장에 미치는 환경요인의 영향)

  • Han, Su Jin;Na, Yeon Ju;Jeon, Da Vine;Kim, Young Sik;Choi, Han Gil;Nam, Ki Wan
    • Ocean and Polar Research
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    • v.38 no.1
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    • pp.47-57
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    • 2016
  • Effects of environmental factors on the zoospore release and germling growth of Chaetomorpha linum were examined. Zoospore release and germling growth experiments were carried out under a combination of temperatures ${\times}$ irradiances, and a single factor of irradiances, salinities, daylength and nutrients (nitrogen, phosphorus). Zoospore release was maximal under condition of $20^{\circ}C{\times}100{\mu}mol\;photons\;m^{-2}s^{-1}$ in a factorial experiment (temperatures ${\times}$ irradiances), at $100{\mu}mol\;photons\;m^{-2}s^{-1}$ in irradiance experiment and at 34 psu in salinity conditions. After 24 days in culture, germling growth was maximal at $25^{\circ}C{\times}100{\mu}mol\;photons\;m^{-2}s^{-1}$ in temperatures ${\times}$ irradiances and at normal seawater (34 psu) in salinity. Germling growth is faster at higher irradiance under same temperature condition and it increased with increasing daylengths (8-16 h). Chaetomorpha linum grew in wide range of N and P concentrations. The growth of germling was maximal at $50{\mu}M\;{PO_4}^{3-}$ and $40{\mu}M\;{NH_4}^+$. Germling growth was more effective under nitrogen addition than phosphorus addition in culture. In conclusion, optimal environmental conditions for zoospore release were $20^{\circ}C$, $100{\mu}mol\;photons\;m^{-2}s^{-1}$, and 34 psu. In addition, the optimal germling growth was observed at $25^{\circ}C$, $100{\mu}mol\;photons\;m^{-2}s^{-1}$, 34 psu, 16 h, $50{\mu}M\;({PO_4}^{3-})$ and $40{\mu}M\;{(NH_4}^+)$.

Individualism and collectivism in ethical decision making (문화성향은 윤리적 의사결정의 과정에 영향을 주는가?)

  • Hong Im Shin
    • Korean Journal of Culture and Social Issue
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    • v.21 no.1
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    • pp.67-96
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    • 2015
  • Do cultural differences affect moral decisions? Two studies were conducted to investigate whether attitudes of individualism vs. collectivism have an impact on ethical decision making. Study 1 (N=92) showed that utilitarianism was preferred in a situation, in which an intervention resulted in the best outcome (i.e., saving more people's lives), while deontology was preferred in a situation, in which the focus was on negative consequences of the intervention (i.e. personal sacrifices). Additionally, there were differences between the idiocentrics and the allocentrics groups regarding morality aspects. In the idiocentrics group, harm and fairness were regarded as more important than other moral aspects, while in the allocentrics group, not only harm and fairness, but also ingroup and authority were perceived as critical moral aspects. In Study 2 (N=30), after lexical decision tasks were conducted for culture priming, the mouse tracking method was used to explore response dynamics of moral decision processes, while judging appropriateness of interventions in moral dilemmas. In Study 2, in a condition, in which the small number of victims were focused upon, there were more maximal deviations and higher Xflips in the individualism priming group than in the collectivism priming group, which showed that the participants in the individualism condition had more deliberative processes before choosing their answers between utilitarianism and deontology. In addition, the participants in the individualism priming condition showed more maximal deviations in the mouse trajectories regarding ingroup related interventions in moral dilemmas than those in the collectivism priming condition. These results implicated the possibilities that the automatic emotional process and the controlled deliberative process in moral decision making might interact with cultural dispositions of the individuals and the focus of situations.

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STudies on the Microbial Pigment(I) (미생물의 색소에 관한 연구. 제1보)

  • Ahn, Tae-Seok;Choi, Yong-Keel;Hong, Soon-Woo
    • Korean Journal of Microbiology
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    • v.15 no.4
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    • pp.159-169
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    • 1977
  • The bacteria of red colonies isolated from soil were identified as Serratia marcescens. The best solvent for pigment extraction was n-buthanol and the pigment was identified as prodigiosene. The extracted pigment was stable on temperature and light but not on acidity. The redpigment color changed into red in alkaline solution. The maximum absorbancy of pigment was 466 nm in alkaline condition and 540 nm in acid condition. And the pigment formed single spot on the TLC(starch). By the result of infra red spectrum, the red pigment has the same absorption pattern comparing with, the prodigisin produced by S. marcescens strain Nima. It was confirmed that the pigment was secondary metabolite and that the maximal peak of production appeared at 30 hrs after the inoculation, when the bacterial growth was in statinary state. Referring to the effect of temperature, the pigment was not formed at $36^{\circ}C$ and the optimal temperature for both of bactrial growth and pigmentation was $30^{\circ}C$. The optimal range of pH for pigmentation was 5.0 and under the condition the bacterial growth was not affected at all. Examining the effects of light, the bacterial pigment ation was more increased in darkness than in visible light.

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Pharmacodynamics of Potassium Channel Opener Lemakalim in the Isolated Rat Hearts and Conscious SHRs under Hyperkalemic and Hypokalemic Condition (칼륨채널 활성물질 Lemakalim의 고칼륨혈증 및 저칼륨혈증에서의 심기능에 대한 영향 및 항 고혈압작용)

  • 신홍섭;신화섭;권광일
    • YAKHAK HOEJI
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    • v.37 no.5
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    • pp.463-475
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    • 1993
  • Pharmacological effects of lemakalim on cardiovascular system were investigated using isolated rat hearts and conscious SHRs subjected to hyperkalemic and hypokalemic condition. In the isolated hearts perfused with normal physiological salt solution(4.7 mM KCI), lemakalim increased cardiac function and coronary flow, and these effects were significantly potentiated under hypokalemic(1.2, 2.5 mM KCI), but attenuated under hyperkalemic(IO mM KCI) condition. In conscious SHRS, lemakalim(0.1, 0.2, 0.3mg/kg, p.o.) produced a dose-related decrease in systolic blood pressure, the maximal hypotensive effect being reached around 0.5 hr after dosing. The intensity and the duration of hypotensive effect of lemakalim were significantly increased when administered in combination with dihydrochlorothiazide (2 mg/kg, p.o.), but decreased with triamterene(32 mg/kg, p.o.). It appears that the differential effects of two types of diuretics on the hypotensive action of lemakalim are due to their hypokalemic and hyperkalemic action, respectively. It is conclued that the concomitant use of $K^{+}$ channel openers and hypokalemic diuretics may be an appropriate model of combination therapy in the treatment of hypertension.

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An Assay Method for Screening Inhibitors of Prolyl 4-hydroxylase in Immortalized Rat Hepatic Stellate HSC-T6 Cells

  • Choi, Hwa-Jung;Soh, Yun-Jo
    • Biomolecules & Therapeutics
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    • v.15 no.4
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    • pp.261-265
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    • 2007
  • Hydroxyproline (HYP) is a post-translational product of proline hydroxylation catalyzed by an enzyme prolyl 4-hydroxylase (P4H) which plays a crucial role in the synthesis of all collagens. Considering the role of collagen and its significance in many clinically important diseases such as liver fibrosis, a great deal of attention has been directed toward the development of an assay at cell-based system. The reason is that cell-based assay system is more efficient than enzyme-based in vitro system and takes much less time than in vivo system. Several assay procedures developed for P4H are laborious, time-consuming and not feasible for the massive-screening. Here, we report the cell-based assay method of prolyl 4-hydroxylase in immortalized rat hepatic stellate HSC-T6 cells. To optimize the cell culture condition to assay for HYP content, various concentrations of reagents were treated for different times in HSC-T6 cells. Our data showed that the treatment with ascorbate in a hypoxic condition for 24 h resulted in the maximal increase of HYP by 1.8 fold. Alternatively, cobalt chloride ($5\;{\mu}M$) and ascorbate ($50\;{\mu}M$) in normoxic states exhibited similar effect on the production of HYP as in a hypoxic condition. Therefore, cobalt chloride can be substituted for a hypoxic condition when an anaerobic chamber is not available. Rosiglitazone and HOE077, known as inhibitors of collagen, synthesis decreased P4H enzyme activity by 32.3% and 15%, respectively, which coincided with previous reports from liver tissues. The level of the smooth muscle ${\alpha}$-actin, a marker of activated stellate cells, was significantly increased under hypoxia, suggesting that our experimental condition could work for screening the anti-fibrotic compounds. The assay procedure took only 3 days after treatment with agents, while assays from the primary stellate cells or liver tissues have taken several weeks. Considering the time and expenses, this assay method could be useful to screen the compounds for the inhibitor of prolyl 4-hydroxylase.