• Title/Summary/Keyword: CelD

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Purification of TGF-$\beta$1 from Bovine Colostrum (젖소 초유로부터 TGF-$\beta$1의 정제)

  • 남명수;배형철;김평현;김완섭;고준수
    • Food Science of Animal Resources
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    • v.22 no.4
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    • pp.343-347
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    • 2002
  • Bovine colostrum contains various bio-functional Proteins. Especially, transforming: growth factor-${\beta}$1 (TGF-${\beta}$1) has a function in concerns with immune response. The purpose of this study was to establish the purification Processing of transforming growth factor-${\beta}$1(TGF-${\beta}$1). The highest concentration of TGF-${\beta}$1 was measured within 48 h after parturition in bovine colostrum using ELISA kit. Purification of TGF-${\beta}$1 from whey protein was carried out by the gel filtration, AF-heparin chromatography and AF-heparin rechromatography. After final purification step, TGF-${\beta}$1 with a molecular weight of 25 kD was obtained, and confirmed by silver staining and western blotting. Finally, TGF-${\beta}$1 was identified native form of 25 kD and reducing form of 12.5 kD by reducing agent.

Estimation of the excavation damage zone in TBM tunnel using large deformation FE analysis

  • Kim, Dohyun;Jeong, Sangseom
    • Geomechanics and Engineering
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    • v.24 no.4
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    • pp.323-335
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    • 2021
  • This paper aims to estimate the range of the excavation damaged zone (EDZ) formation caused by the tunnel boring machine (TBM) advancement through dynamic three-dimensional large deformation finite element analysis. Large deformation analysis based on Coupled Eulerian-Lagrangian (CEL) analysis is used to accurately simulate the behavior during TBM excavation. The analysis model is verified based on numerous test results reported in the literature. The range of the formed EDZ will be suggested as a boundary under various conditions - different tunnel diameter, tunnel depth, and rock type. Moreover, evaluation of the integrity of the tunnel structure during excavation has been carried out. Based on the numerical results, the apparent boundary of the EDZ is shown to within the range of 0.7D (D: tunnel diameter) around the excavation surface. Through series of numerical computation, it is clear that for the rock of with higher rock mass rating (RMR) grade (close to 1st grade), the EDZ around the tunnel tends to increase. The size of the EDZ is found to be direct proportional to the tunnel diameter, whereas the depth of the tunnel is inversely proportional to the magnitude of the EDZ. However, the relationship between the formation of the EDZ and the stability of the tunnel was not found to be consistent. In case where the TBM excavation is carried out in hard rock or rock under high confinement (excavation under greater depth), large range of the EDZ may be formed, but less strain occurs along the excavation surface during excavation and is found to be more stable.

Development of Isolation and Cultivation Method for Outer Root Sheath Cells from Human Hair Follicle and Construction of Bioartificial Skin

  • Seo, Young-Kwon;Lee, Doo-Hoon;Shin, Youn-Ho;You, Bo-Young;Lee, Kyung-Mi;Song, Key-Yong;Seo, Seong-Jun;Whang, Sung-Joo;Kim, Young-Jin;Park, Chang-Seo;Chang, Ij-Seop;Park, Jung-Keug
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.2
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    • pp.151-157
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    • 2003
  • Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS tells is 2.↑ $\times$ 10$^3$cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA(0.02%) solution for 15 min at 37$^{\circ}C$, however, our modified method was able to obtain about 6.9 $\times$ 10$^3$cel1s/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 $\times$ 10$^{7}$ cells was obtained in a serum-free medium. while a modified E-medium with mitomycin C-treated feeder tells produced a total of 6.3 $\times$ 10$^{7}$ Cel1s over 17 days When Starting With 7.5 $\times$ 10$^4$cells. Finally, We Confirmed the effectiveness of our ORS tell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro

A Proposed Analytical Model for the Debris Flow with Erosion and Entrainment of Soil Layer (지반의 침식 및 연행작용을 고려한 토석류 해석 모델 제안)

  • Lee, Kwang-Woo;Park, Hyun-Do;Jeong, Sang-Seom
    • Journal of the Korean Geotechnical Society
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    • v.32 no.10
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    • pp.17-29
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    • 2016
  • A debris flow analysis model has been developed to simulate the erosion and entrainment of soil layer. Special attention is given to the model which represents strength softening behaviour of soil layer due to velocity of deformation. The 3D FE analysis by Coupled Eulerian-Lagrangian (CEL) model is conducted to simulate the debris flow. The model is validated using published data on laboratory experiment (Mangeny et al., 2010). It has been definitely shown that proposed model is in good agreement with the results of laboratory data. Futhermore, the FE analysis is conducted to ensure capability of simulating the real scale debris flow. The result of Ramian watershed, Korea shows that the debris flow has increased the volume and speed and it is in good agreement with field investigation. Based on this, it is confirmed that proposed model shows good agreement of the behavior of the actual and analytical debris flow.

Numerical Analysis for Comparing Beam-spring and Continuum Model for Buried Pipes Considering Soil-pipe Interaction (매설관과 지반의 상호작용을 고려한 보-스프링 모델과 연속체 모델의 수치해석적 비교 연구)

  • Jeonghun Yang;Youngjin Shin;Hangseok Choi
    • Journal of the Korean GEO-environmental Society
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    • v.24 no.9
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    • pp.15-24
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    • 2023
  • The behavior of buried pipes is directly influenced by the nonlinearity and complex characteristics of the surrounding soil. However, the simplified beam-spring model, which ignores the nonlinearity and complex behavior of soil, is commonly used in practice. In response, several studies have employed continuum analysis methods to account for the nonlinear and complex behavior of the soil. This paper presents various numerical continuum analysis techniques and verifies their comparison with full-scale tests. The study found that reaction force results close to the full-scale test could be obtained by applying contact surface characteristics that take into account the interaction between the ground and the buried pipe. In the case of sharing pipe and soil node method and ignoring the interaction between pipe and soil, excessive reaction force was derived, and the failure shapes were different. In addition, this study applied the dynamic explicit analysis method, ALE method, and CEL method. It was confirmed that the displacement-reaction relationship and failure shape are similar to those of the static analysis.

Development of Recombinant Chinese Hamster Ovary Cell Lines Producing Human Thrombopoietin or Its Analog

  • Chung, Joo-Young;Ahn, Hae-Kyung;Lim, Seung-Wook;Sung, Yun-Hee;Koh, Yeo-Wook;Park, Seung-Kook;Lee, Gyun-Min
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.759-766
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    • 2003
  • Recombinant Chinese hamster ovary (rCHO) cell lines expressing a high level of human thrombopoietin (hTPO) or its analog, TPO33r, were obtained by transfecting expression vectors into dihydrofolate reductase-deficient (dhfr) CHO cells and subsequent gene amplification in media containing stepwise increments in methotrexate (MTX) level such as 20, 80, and 320 nM. The parental clones with a hTPO expression level $>0.40\;{\mu}g/ml$ (27 out of 1,200 clones) and the parental clones with a TPO33r expression level $>0.20\;{\mu}g/ml$ (36 out of 400 clones) were subjected to 20 nM MTX. The clones that displayed an increased expression level at 20 nM MTX were subjected to stepwise increasing levels of MTX such as 80 and 320 nM. When subjected to 320 nM MTX, most clones did not display an increased expression level, since the detrimental effect of gene amplification on growth reduction outweighed its beneficial effect of specific TPO productivity ($q_{TPO}$) enhancement at 320 nM MTX. Accordingly, the highest producer subclones ($1-434-80^{*}$ for hTPO and $2-3-80^{*}$ for TPO33r), whose $q_{TPO}$ was 2- to 3-fold higher than that of their parental clones selected at 80 nM MTX, were isolated by limiting dilution method and were established as rCHO cel1 lines. The $q_{TPO}$ of $1-434-80^{*}\;and\;2-3-80^{*}\;was\;5.89{\pm}074\;and\;1.02{\pm}0.23\;{\mu}g/10^6$ cells/day, respectively. Southern and Northern blot analyses showed that the enhanced $q_{TPO}$ of established rCHO cell lines resulted mainly from the increased TPO gene copy number and subsequent increased TPO mRNA level. The hTPO and TPO33r produced from the established rCHO cell lines were biologically active in vivo, as demonstrated by their ability to elevate platelet counts in treated mice.

Production of Single-Cell Protein from Methanol (Part 2). Batch Fermentation and Continuous Fermentation by Methylomonas methanolica (Methanol을 이용한 단세포단백질 생산에 관한 연구 (제 2 보), Methylomons methanolica YUFE 101의 회분발효와 연속발효)

  • 유주현;정건섭;변유량
    • Microbiology and Biotechnology Letters
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    • v.7 no.2
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    • pp.71-74
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    • 1979
  • The growth characteristics of Methylomonas methanolica YUFE 101, isolated from sewage samples, have been studied. conclusions of the study were; (1) Optimum cultivation pH and temperature are 6.3 and 32.5$^{\circ}C$ respectively (2) The specific oxygen uptake rate was 332 $\mu$ι/mg-dry weight/hr. (3) The maximum specific growth rate was 0.19 h $r^{-1}$ and celluar yield was 0.43 g-dry cell/g-methanol in batch culture, (4) The maximum biomass productivity achieved was 0.21 g-dry cel1/ι/hr at a dillution rate of 0.1 h $r^{-1}$ during continuous cultivation. (5) The contents of crude protein and total nucleic acid in the dry cell were 73 % and 12 % respectively.

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Damage rate assessment of cantilever RC walls with backfill soil using coupled Lagrangian-Eulerian simulation

  • Javad Tahamtan;Majid Gholhaki;Iman Najjarbashi;Abdullah Hossaini;Hamid Pirmoghan
    • Geomechanics and Engineering
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    • v.36 no.3
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    • pp.231-245
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    • 2024
  • In recent decades, the protection and vulnerability of civil structures under explosion loads became a critical issue in terms of security, which may cause loss of lives and structural damage. Concrete retaining walls also restrict soils and slopes from displacements; meanwhile, intensive temporary loading may cause massive damage. In the current study, the modified Johnson-Holmquist (also known as J-H2) material model is implemented for concrete materials to model damages into the ABAQUS through user-subroutines to predict the blasting-induced concrete damages and volume strains. For this purpose, a 3D finite-element model of the concrete retaining wall was conducted in coupled Eulerian-Lagrangian simulation. Subsequently, a blast load equal to 500 kg of TNT was considered in three different positions due to UFC 3-340-02. Influences of the critical parameters in smooth blastings, such as distance from a free face, position, and effective blasting time, on concrete damage rate and destroy patterns, are explored. According to the simulation results, the concrete penetration pattern at the same distance is significantly influenced by the density of the progress environment. The result reveals that the progress of waves and the intensity of damages in free-air blasting is entirely different from those that progress in a dense surrounding atmosphere such as soil. Half-damaged elements in air blasts are more than those of embedded explosions, but dense environments such as soil impose much more pressure in a limited zone and cause more destruction in retaining walls.

Stylized Specular Reflections Using Projective Textures based on Principal Curvature Analysis (주곡률 해석 기반의 투영 텍스처를 이용한 스타일 반사 효과)

  • Lee, Hwan-Jik;Choi, Jung-Ju
    • Journal of the HCI Society of Korea
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    • v.1 no.1
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    • pp.37-44
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    • 2006
  • Specular reflections provide the visual feedback that describes the material type of an object, its local shape, and lighting environment. In photorealistic rendering, there have been a number of research available to render specular reflections effectively based on a local reflection model. In traditional cel animations and cartoons, specular reflections plays important role in representing artistic intentions for an object and its related environment reflections, so the shapes of highlights are quite stylistic. In this paper, we present a method to render and control stylized specular reflections using projective textures based on principal curvature analysis. Specifying a texture as a pattern of a highlight and projecting the texture on the specular region of a given 3D model, we can obtain a stylized representation of specular reflections. For a given polygonal model, a view point, and a light source, we first find the maximum specular intensity point, and then locate the texture projector along the line parallel to the normal vector and passing through the point. The orientation of the projector is determined by the principal directions at the point. Finally, the size of the projection frustum is determined by the principal curvatures corresponding to the principal directions. The proposed method can control the position, orientation, and size of the specular reflection efficiently by translating the projector along the principal directions, rotating the projector about the normal vector, and scaling the principal curvatures, respectively. The method is be applicable to real-time applications such as cartoon style 3D games. We implement the method by Microsoft DirectX 9.0c SDK and programmable vertex/pixel shaders on Nvidia GeForce FX 7800 graphics subsystems. According to our experimental results, we can render and control the stylized specular reflections for a 3D model of several ten thousands of triangles in real-time.

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Identification and Characterization of Protein Encoded by orf382 as $\small{L}$-Threonine Dehydrogenase

  • Ma, Fei;Wang, Tianwen;Ma, Xingyuan;Wang, Ping
    • Journal of Microbiology and Biotechnology
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    • v.24 no.6
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    • pp.748-755
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    • 2014
  • In the genome annotation of Escherichia coli MG1655, the orf382 (1,149 bp) is designated as a gene encoding an alcohol dehydrogenase that may be Fe-dependent. In this study, the gene was amplified from the genome by PCR and overexpressed in Escherichia coli BL21(DE3). The recombinant $6{\times}$His-tag protein was then purified and characterized. In an enzymatic assay using different hydroxyl-containing substrates (n-butanol, $\small{L}$-threonine, ethanol, isopropanol, glucose, glycerol, $\small{L}$-serine, lactic acid, citric acid, methanol, or $\small{D}$-threonine), the enzyme showed the highest activity on $\small{L}$-threonine. Characterization of the mutant constructed using gene knockout of the orf382 also implied the function of the enzyme in the metabolism of $\small{L}$-threonine into glycine. Considering the presence of tested substrates in living E. coli cel ls and previous literature, we believed that the suitable nomenclature for the enzyme should be an $\small{L}$-threonine dehydrogenase (LTDH). When using $\small{L}$-threonine as the substrate, the enzyme exhibited the best catalytic performance at $39^{\circ}C$ and pH 9.8 with $NAD^+$ as the cofactor. The determination of the Km values towards $\small{L}$-threonine (Km = $11.29{\mu}M$), ethanol ($222.5{\mu}M$), and n-butanol ($8.02{\mu}M$) also confirmed the enzyme as an LTDH. Furthermore, the LTDH was shown to be an ion-containing protein based on inductively coupled plasma-atomic emission spectrometry with an isoelectronic point of pH 5.4. Moreover, a circular dichroism analysis revealed that the metal ion was structurally and enzymatically essential, as its deprivation remarkably changed the ${\alpha}$-helix percentage (from 12.6% to 6.3%).