• Title/Summary/Keyword: Transient model

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Effects of Mutagenesis for Glycosylation Sites of Recombinant Human EPO During Production from Cultured CHO Cell

  • Lee, Hyun-Gi;Seong, Hwan-Hoo;Im, Seok-Ki;Chung, Hee-Kyoung;Lee, Poongyeon;Lee, Yeun-Kun;Min, Kwan-Sik;Chang, Won-Kyoung;Lee, Hoon-Taek
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.97-97
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    • 2002
  • Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.

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Three-Dimensional Numerical Simulation of Impacts of Urbanization on Groundwater Flow and Salt Transport in a Coastal Aquifer, Suyeong-Gu, Busan, Korea (한국 부산광역시 수영구 지역 해안 대수층 내의 지하수 유동 및 염분 이동에 대한 도시화의 영향 삼차원 수치 모의)

  • Cho, Hyeon-Jo;Kim, Jun-Mo
    • Journal of Soil and Groundwater Environment
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    • v.14 no.6
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    • pp.1-18
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    • 2009
  • A series of three-dimensional numerical simulations using a generalized multidimensional hydrodynamic dispersion numerical model is performed to simulate effectively and to evaluate quantitatively impacts of urbanization on density-dependent groundwater flow and salt transport in a coastal aquifer system, Suyeong-Gu, Busan, Korea. A series of steady-state numerical simulations of groundwater flow and salt transport before urbanization with material properties of geologic formations, which are established by numerical modeling calibrations considering all the urbanization factors, is performed first without considering all the urbanization factors. A series of transient-state numerical simulations of groundwater flow and salt transport after urbanization is then performed considering the urbanization factors individually and all together. Finally, the results of both numerical simulations are compared with each other and analyzed. The results of the numerical simulations show that density-dependent groundwater flow, salt transport, and seawater intrusion in the coastal aquifer system are intensively and extensively impacted by the urbanization factors. Especially, these urbanization factors result in the changes of the total groundwater volume and salt mass in the coastal aquifer system. However, such impacts of each urbanization factor are not spatially uniform but locally different.

Neuroprotective effects of herbal mixture HT070 on global cerebral ischemia in rats

  • Song, Jungbin;Lee, Donghun;Kim, Young-Sik;Lee, Hyun Jeong;Lee, Seunggyeong;Kim, Dong Kuk;Kang, Shin Ho;Shin, Yong Kook;Choi, Ho-Young;Kim, Hocheol
    • The Korea Journal of Herbology
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    • v.31 no.4
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    • pp.101-109
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    • 2016
  • Objectives : HT070 is a mixture of herbal extracts from root of Scutellaria baicalensis and stem bark of Eleutherococcus senticosus , which have long been used for stroke therapy in traditional Korean Medicine. The purpose of this study was to investigate the neuroprotective effects of HT070 on global cerebral ischemia and its potential mechanisms.Methods : Transient global cerebral ischemia was produced by 10 min of four-vessel occlusion (4-VO) in male Wistar rats. HT070 was administered orally at a dosage of 200 mg/kg twice at 0 and 90 min after reperfusion. Hippocampal neuronal damage was measured 7 days after reperfusion. To explore the potential mechanisms, we used hydrogen peroxide (H2O2)-induced rat pheochromocytoma (PC12) cells as an in vitro model. PC12 cells were pretreated with HT070 for 1 h and then exposed to 100 μM H2O2 for 6 h in the presence of HT070. Cell viability was measured by MTT assay and the mRNA expression of Bax, Bcl-2, iNOS and COX-2 were measured by quantitative RT-PCR.Results : Oral administration of HT070 at a dose of 200 mg/kg significantly reduced neuronal death in the hippocampal CA1 region by 13.4% as compared to the vehicle-treated group. HT070 increased cell viability, reversed the down-regulated Bcl-2 mRNA level, and suppressed the up-regulated mRNA expressions of Bax, iNOS, and COX-2 in H2O2-treated PC12 cells.Conclusions : HT070 protects against delayed neuronal death after global cerebral ischemia and its neuroprotection properties might be attributed to the inhibition of mitochondrial apoptosis and ROS-generating enzymes.

Effect of Gd-DTPA on Diffusion in Canine Brain with Hyperacute Stroke (초급성 뇌경색을 일으킨 개에서 Gd-조영제의 주입이 뇌의 확산에 미치는 영향)

  • 김범수;정소령;신경섭
    • Investigative Magnetic Resonance Imaging
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    • v.6 no.2
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    • pp.158-165
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    • 2002
  • Purpose : To evaluate the effect of Gd-DTPA on signal intensity of diffusion-weighted magnetic resonance(MR) image and apparent diffuse coefficient (ADC) in dog brain with hype racute stroke. Materials and methods : Experimental canine model of hyperacute cerebral infarction was made by selective intraarterial embolization with particulate embolic material. Diffusion-weighted MR imaging was performed in five dogs at 1 hour after the embolization of internal carotid artery. After intravenous bolus injection of Gd- DTPA, additional 11 diffusion-weighted MR images were serially obtained from 2 minutes to 90 minutes after injection in each dog. The author evaluated findings of hyperacute cerebral infarction on diffusion-weighted MR imaging, and calculated mean signal intensity and mean ADC in infarcted region and contralateral normal region. Statistical analysis of mean signal intensity, mean ADC and contrast-noise ratio before and after Gd-DTPA injection was performed. Results : Hyperacute cerebral infarction developed in all five dogs on diffusion-weighted MR images obtained 1 hour after embolization. The area of hyperacute infarction had steady increase in signal intensity on diffusion-weighted MR image and decrease in ADC. In normal perfusion area, decrease in signal intensity was observed at 2 minutes the Gd-DTPA injection, whereas ADC did not changed. Conclusion : Intravenous injection of Gd-DTPA had no influence on ADC in both hyperacute infarction and normally perfused are a, but caused initial transient signal reduction in normally perfused area on diffusion-weighted MR image due to susceptibility effect of Gd-DTPA. It is important to calculate ADC in evaluating the effect of diffusion after injection of Gd-DTPA.

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A Non-consecutive Cloth Draping Simulation Algorithm using Conjugate Harmonic Functions (켤레조화함수를 이용한 비순차적 의류 주름 모사 알고리즘)

  • Kang Moon Koo
    • Journal of KIISE:Software and Applications
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    • v.32 no.3
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    • pp.181-191
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    • 2005
  • This article describes a simplified mathematical model and the relevant numerical algorithm to simulate the draped cloth on virtual human body. The proposed algorithm incorporates an elliptical, or non-consecutive, method to simulate the cloth wrinkles on moving bodies without resorting to the result of the past time-steps of drape simulation. A global-local analysis technique was employed to decompose the drape of cloths into the global deformation and the local wrinkles that will be superposed linearly The global deformation is determined directly by the rotation and the translation of body parts to generate a wrinkle-free yet globally deformed shape of cloth. The local wrinkles are calculated by solving simple elliptical equations based on the orthogonality between conjugate harmonic functions representing the wrinkle amplitude and the direction of wrinkles. The proposed method requires no interpolative time frames even for discontinuous body postures. Standing away from the incremental approach of time integration in conventional methods, the proposed method yields a remarkable reduction of CPU time and an enhanced stability. Also, the transient motion of cloth could be achieved by interpolating between the deformations corresponding to each static posture.

The Removal of Toluene by a Granular Activated Carbon Bioreactor using Yeast (Yeast와 입상활성탄을 이용한 미생물반응기의 휘발성유기화합물 분해 특성)

  • NamGung, Hyeong-Kyu;Shin, Seung-Kyu;Ahmed, Zubair;Song, Ji-Hyeon
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.12
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    • pp.1218-1224
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    • 2008
  • A liquid culture of yeast "Candida tropicalis" was used in a fluidized bioreactor to achieve high removal efficiencies of volatile organic compounds (VOCs). In this study, granular activated carbon (GAC) was used as a fluidized material to improve adsorptive capacity as well as mass transfer of gaseous toluene, the model VOC. The GAC fluidized bioreactor demonstrated toluene removal efficiencies ranging from 50 to 80%, when inlet toluene loading varied in a range between 13.1 and 37.4 g/m$^3$-hr. The maximum elimination capacity determined in the GAC fluidized bioreactor was 172 g/m$^3$-hr at a toluene loading of 291 g/m$^3$-hr. Transient loading experiments revealed that the removal efficiency was remained unchanged during an increased loading period, and toluene introduced to the bioreactor was first absorbed to GAC and then slowly desorbed and became available to the yeast culture. Hence the fluidized GAC helped to achieve an improved mass transfer between the gas and liquid phases, resulting in high toluene removal capacity. Consequently, the GAC fluidized bioreactor using C. tropicalis can be successfully applied for the removal of VOCs, and is a feasible alternative over conventional processes such as packed-bed biofilters.

Temperature-Induced Stresses and Deformation in Composite Box Girder Bridges (합성 박스형 교량의 온도에 의한 응력 및 변형)

  • Chang, Sung Pil;Im, Chang Kyun
    • Journal of Korean Society of Steel Construction
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    • v.9 no.4 s.33
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    • pp.659-672
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    • 1997
  • Thermal response induced from nonlinear temperature distribution in composite box gilder bridges depends on several variables(environmental conditions, physical and material properties, location and orientation of bridge, and cross-section geometry). In this paper, parametric study are conducted in order to find the effects of variations of seasons, location and orientation of bridge, sectional geometry and some material properties on the axial deformation, curvature and stresses in composite box girder bridge. A two-dimensional transient finite element model to conduct this parametric studies is briefly presented. Firstly, the effects of the parameters on the diurnal variation of curvature are considered, and for the time of maximum curvature, on the distribution of temperature and stresses of composite box girder sectional are considered. Finally, some considerations about the influence of the parameters on the daily maximum values of axial deformation, curvature and stresses are carried out. The influence of thermal effect on structures is important as much as the influence of live or dead load in some cases. In the design of steel composite bridges, the thermal stresses calculated on the supposition that the temperature difference between the concrete slab and steel girder is $10^{\circ}C$ and the temperature distributions are uniform in concrete slab and steel girder can be underestimated.

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Avoidance of Internal Resonances in Hemispherical Resonator Assemblies from Fused Quartz Connected by Indium Solder

  • Sarapuloff, Sergii A.;Rhee, Huinam;Park, Sang-Jin
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2013.04a
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    • pp.835-841
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    • 2013
  • Modern solid-state gyroscopes (HRG) with hemispherical resonators from high-purity quartz glass and special surface superfinishing and ultrathin gold coating become the best instruments for precise-grade inertial reference units (IRU) targeting long-term space missions. Designing of these sensors could be a notable contribution into development of Korea as a space nation. In participial, 40mm diameter thin-shell resonator from high-purity fused quartz, fabricated as a single-piece with its supporting stem has been designed, machined, etched, tuned, tested, and delivered by STM Co. (ATS of Ukraine) several years ago; an extremely-high Q-factor (upto 10~20 millions) has been shown. Understanding of the best way how to match such a unique sensor with inner glass assembly of the gyro means how to use the high potential in a maximal extent; and this has become the urgent task. Inner quartz glass assembly has a very thin indium (In) layer soldered the resonator and its silica base (case), but effects of internal resonances between operational modal pair of the shell-cup and its side (parasitic) modes can notable degrade the potential of the sensor as a whole, instead of so low level of resonator's intrinsic losses. Unfortunately, there are special combinations of dimensions of the parts (so-called, "resonant sizes"), when intensive losses of energy occurs. The authors proposed to use the length of stem's fixture as an additional design parameter to avoid such cases. So-called, a cyclic scheme of finite element method (FEM) and ANSYS software were employed to estimate different combinations of gyro assembly parameters. This variant has no mismatches of numerical origin due to FEM's discrete mesh. The optimum length and dangerous "resonant lengths" have been found. The special attention has been paid to analyses of 3D effects in a cup-stem transient zone, including determination of a difference between the positions of geometrical Pole of the resonant hemisphere and of its "dynamical Pole", i.e., its real zone of oscillation node. Boundary effects between the shell (cup) and 3D short "beams" (inner and outer stems) have been ranged. The results of the numerical experiments have been compared with the classic model of a quasi-hemispherical shell band with inextensional midsurface, and the solution using Rayleigh's functions of the $1^{st}$ and $2^{nd}$ kinds. To guarantee the truth of the recommended sizes to a designer of the real device, the analytical and FEM results have been compared with experimental data for a party of real resonators. The consistency of the results obtained by different means has been shown with errors less than 5%. The results notably differ from the data published earlier by different researchers.

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The Change of c-jun Promoter Activity in TPA-Induced U937 Cells Infected with Human Cytomegalovirus (HCMV) (TPA로 분화된 U937 세포에서 사람 세포거대바이러스에 의한 c-jun Promoter 활성도의 변화)

  • Park, Chung-Gyu;Kim, Dae-Joong;Kim, Jin-Hee;Han, Tae-Hee;Hwan, Eung-Soo;Choi, Myong-Sik;Kook, Yoon-Hoh;Choi, Sung-Bae;Cha, Chang-Yong
    • The Journal of Korean Society of Virology
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    • v.29 no.2
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    • pp.129-136
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    • 1999
  • Transient transfection assay has been done to evaluate whether the c-jun activation would be prerequisite to the induction of permissiveness against human cytomegalovirus using in vitro cell model in which U937 has been induced to express CD11b and CD14 to become potential monocyte/macrophage cells by TPA treatment. U937 cells were treated with $10\;{\mu}M$, $50\;{\mu}M$ or $100\;{\mu}M$ of TPA. The cell morphology change was observed and the expression of the CD11b and CD14 was confirmed by FACS. Differentiated cells were transfected with pJLuc reporter vector which contained the wild type murine c-jun promoter spanning the SP1, CTF, ATF/CREB and MEF-2 binding sites upstream of the firefly luciferase gene. After 48 hrs of transfection, the cells were infected with HCMV Towne strain and the luciferase activity was assessed at 1 hand 4 h pi. The transfection assay showed no activation of the c-jun promoter at 1 h pi, instead, it showed 2 times increase of the its activity at 4 h pi. There was no difference of the c-jun promoter activation between TPA treated and untreated U937 cells, implying that c-jun activation might not be prerequisite for allowing cells to be premissive to HCMV, although HCMV infection itself could activate c-jun promoter.

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A study of the Mori Radicis Cortex pre-treatment on transient ischemic brain injury in mice (상백피(桑白皮) 메탄올 추출물 전처치가 일과성 허혈에 의한 생쥐의 뇌 손상에 미치는 영향)

  • Chung, Byung-Woo;Lim, Jae-Yu;Lee, Se-Eun;Lee, Byoungho;Lim, Sehyun;Lim, Chiyeon;Cho, Suin
    • The Korea Journal of Herbology
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    • v.32 no.1
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    • pp.25-31
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    • 2017
  • Objectives : Mori Radicis Cortex (MRC), the root epidermis of Morus alba L., has been traditionally used to treat lung-related diseases in Korean Medicine. The common of MRC is Mulberry bark Morus bark, and it's pharmaceutical properties and taste are known as sweet and cold, and it promotes urination and reduce edema by reducing heat from the lungs and soothe asthma. In the present study, anti-apoptotic mechanism of MRC in middle cerebral artery occlusion (MCAO) model in mice. Methods : Two-hundred grams of MRC was extracted with methanol at room temperature for 5 days, and this was repeated one time. After filtration, the methanol was removed using vacuum evaporator, then stored at $-20^{\circ}C$ until use. C57BL/6 male mice were housed in an environment with controlled humidity, temperature, and light cycle. In order to determine beneficial effects of MRC on ischemia induced brain damage, infarct volume, neurological deficit scores, activities of several apoptosis-related proteins such as caspase-8, -9, Bcl-xL in MCAO-induced brains of mice were analyzed. Mice in MRC-treated groups were orally administered 30, 100, or 300 mg/kg of body weight for three consecutive days before commencing the MCAO procedure. Results : Pre-treatment of MRC significantly reduced infarct volume in MCAO subjected mice applied with 300 mg/kg of MRC methanol extract, and MRC effectively inhibited Bcl-xL reduction and caspase-9 activation caused by MCAO-induced brain damage. Conclusions : MRC showed neuro-protective effects by regulating apoptosis-related protein signals, and it can be a potential candidate for the therapy of ischemia-induced brain damage.