• Title/Summary/Keyword: 3D Expression Method

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Introduction of VP6 Gene into Potato Plant by Agrobacterium-mediated Transformation and Analysis of VP6 Expression in Transgenic Potatoes (Rotavirus VP6 유전자의 감자식물체내로의 도입과 형질전환체의 발현분석)

  • Youm, Jung-Won;Jeon, Jae-Heung;Jung, Jae-Yeol;Lee, Byoung-Chan;Kang, Won-Jin;Kim, Mi-Sun;Kim, Chul-Joong;Joung, Hyouk;Kim, Hyun-Soon
    • Journal of Plant Biotechnology
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    • v.29 no.2
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    • pp.93-98
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    • 2002
  • A VP6 fragments was subcloned with BamHI in the binary pMBP-1 vector under Califlower Mosaic Virus (CaMV) 355 promoter and neomycin phosphotransferase II (npt II) gene. The recombinant binary vector was mobilized into Agrobacterium-tumefaciens LBA4404 by the freeze-thaw method and potato (Solanum tubensum L. cv Desiree) was transformed by modified leaf-disc cocultivation. Shoots were induced on MS medium with 0.01 mg/L NAA, 0.1 mg/L GA$_3$, 2.0 mg/L Zeatin, 100.0 mg/L kanamycin, 500.0 mg/L carbenicillin. In order to identify the copy number of VP6 into potato plant, total genomic DNA was isolated from transgenic potato and analysed by Southern blotting. Genomic DNA and total mRNA analysis demonstrated the incorporation of the foreign gene into the potato genome, as well as their transcription.

Effects of Anabolic Steroids of Pork on Proliferation and Differentiation of Myogenic Satellite Cell (돼지 고기의 아나볼릭 스테로이드가 Myogenic Satellite Cell의 증식과 분화에 미치는 영향)

  • Lee, Dong-Mok;Lee, Ki-Ho;Cheon, Yong-Pil;Chun, Tae-Hoon;Choi, In-Ho
    • Food Science of Animal Resources
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    • v.30 no.5
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    • pp.842-850
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    • 2010
  • Sex steroids are known to be involved in skeletal muscle development (anabolic effect) and are frequently used in medicines. It has been known that pork contains a variety of steroids that are mainly synthesized in the gonads (testis and ovary). Thus, the present study was conducted to evaluate the effects of anabolic steroids of pork on the proliferation and differentiation of myogenic satellite cells (MSC). Three different methods (M1, M2, and M3) were developed for the isolation and purification of steroids from porcine tissues. Among three extraction methods that we developed, M3 was the best method with respect to the quantities of steroids and the induction of MSC proliferation. Hormonal analysis showed that the steroid hormone levels were the highest in muscle and fat of intact male than those of castrated males and females. In addition, the highest serum levels of nandrolone and testosterone were detected in intact males, whereas estrone and $17{\beta}$-estradiol levels were similar in the entire experimental serum samples. Expression of androgen receptor (AR), myoD, desmin, and myogenin in bovine muscle cells were significantly up-regulated by the treatment of steroid extracts. The highest increas of myogenin and AR mRNA abundance were observed in the MSCs treated with M3 extract (p<0.001). Altogether, the present research showed the positive effect of steroids on MSC proliferation and differentiation in vitro. These results would certainly imply a beneficial effect of pork consumption on human muscle development.

Enhancement of the solubility of human tissue inhibitor of matrix metallocroteinase-2 (TIMP-2) in E. coli using a modified in vitro mutagenesis (새로운 유전자 재조합 방법을 이용한 대장균에서의 인간 tissue inhibitor of mtrix metalloproteinase-2 (TIMP-2) 유전자의 가용성 발현)

  • Kim, Jong-Uk;Choi, Dong-Soon;Joo, Hyun;Min, Churl-K.
    • KSBB Journal
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    • v.23 no.3
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    • pp.231-238
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    • 2008
  • The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

Biological Activity of Mixed Extracts of Acanthopanax senticosus and Citrus unshiu Fermented with Bovista plumbea for Inner Beauty (이너뷰티 소재로서의 생물전환된 가시오가피-진피 혼합 추출물의 생물학적 활성)

  • Eun Jeong Kim;So Yeon Kim;Su Yeon Kang;Yung Choon Yoo;Taek Joon Yoon;Gye Won Lee;Young Ho Cho
    • Journal of Life Science
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    • v.33 no.7
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    • pp.555-564
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    • 2023
  • In this study, the anti-oxidant activity, elastase inhibitory activity, and skin moisturizing effect of mixed extracts of Acanthopanax senticosus and Citrus unshiu fermented with Bovista plumbea (B-MEAC) were evaluated to verify the availability as a material for inner beauty. The DPPH radical scavenging activity of B-MEAC was showed in a dose-dependent manner (SC50=156.1±0.82 ㎍/ml). Also, B-MEAC inhibited the elastase activity in a concentration-dependent manner (p<0.001). To study the effect of B-MEAC on mouse skin hydration, skin moisture content and transepidermal water loss (TEWL) measured. As a result, skin moisture content increased (p<0.001) and TEWL decreased (p<0.01) compared to the dry-induced control group. The effect on the change of collagen fibers in the dry-induced mouse skin was examined through Masson's trichrome staining. In the group administered with B-MEAC, the amount of collagen relatively increased compared to the control group, and the intensity of blue color increased. The effect on the moisturizing function of the dry-induced mouse skin was examined by Western blot method. In the group administered with B-MEAC, the expression of matrix metalloproteinase-1 (MMP-1) protein decreased compared to the control group. In addition, the expression level of collagen1A1 (COL1A1), hyaluronan synthase-2 (HAS2), filaggrin, and aquaporin-3 (AQP3) recovered (p<0.001). Therefore, these results suggest the potential of B-MEAC as a skin hydration agent for inner beauty.

Stress distribution following face mask application using different finite element models according to Hounsfield unit values in CT images (CT상의 HU 수치에 따른 상악골 전방견인 효과의 유한요소 분석)

  • Chung, Dong-Hwa
    • The korean journal of orthodontics
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    • v.36 no.6
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    • pp.412-421
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    • 2006
  • Objective: The result of finite element analysis depends on material properties, structural expression, density of element, and boundar or loading conditions. To represent proper elastic behavior, a finite element model was made using Hounsfield unit (HU) values in CT images. Methods: A 13 year 6 month old male was used as the subject. A 3 dimensional visualizing program, Mimics, was used to build a 3D object from the DICOM file which was acquired from the CT images. Model 1 was established by giving 24 material properties according to HU. Model 2 was constructed by the conventional method which provides 2 material properties. Protraction force of 500g was applied at a 45 degree downward angle from Frankfort horizontal (FH) plane. Results: Model 1 showed a more flexible response on the first premolar region which had more forward and downward movement of the maxillary anterior segment. Maxilla was bent on the sagittal plane and frontal plane. Model 2 revealed less movement in all directions. It moved downward on the anterior part and upward on the posterior part, which is clockwise rotation of the maxilla. Conclusion: These results signify that different outcomes of finite element analysis can occur according to the given material properties and it is recommended to use HU values for more accurate results.

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

  • Choi, Kwang-Hwan;Yoon, Ji Won;Kim, Minsu;Jeong, Jinsol;Ryu, Minkyung;Park, Sungkwon;Jo, Cheorun;Lee, Chang-Kyu
    • Food Science of Animal Resources
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    • v.40 no.4
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    • pp.659-667
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    • 2020
  • Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro. This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.

The Cytotoxic Effect of Oral Wet Wipes on Gingival Cells (시판 중인 구강청결티슈의 세포 독성 관찰)

  • Jung, Im-hee;Park, Ji Hyeon;Lee, Min Kyeng;Hwang, Young Sun
    • Journal of dental hygiene science
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    • v.18 no.2
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    • pp.76-84
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    • 2018
  • Wet wipes are being increasingly used because of their convenience. Particularly, oral wet wipes are useful for regular cleaning of a baby's mouth after birth. Therefore, the consumption of oral wet wipes has increased over the past few years and a variety of products are commercially available. However, product information on safety is not sufficiently provided and still raises doubts regarding adverse effects. To confirm the safety of wet wipes as an oral hygiene item and provide information for their use, we investigated the cytotoxicity of oral wet wipes and verified the underlying mechanism. The anti-bacterial effect of oral wet wipes was analyzed using the disk diffusion method. The cytotoxic effects of oral wet wipes were observed based on morphological changes using microscopy and determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in gingival epithelial cells and gingival fibroblasts. Evaluation of apoptosis by oral wet wipes was explored using propidium iodide flow cytometric analysis and a terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL) assay. Apoptosis-related molecules were also analyzed using western blotting. Five types of oral wet wipes were tested, and two products from Fisher-Price and Dr. Kennedy revealed strong cytotoxic effects on gingiva epithelial cells and gingiva fibroblasts, although they also showed intense anti-bacterial effects on oral bacteria. Cell cycle arrest in the G2/M phase and apoptosis were observed based on treatment of extracts from Fisher-Price and Dr. KENNEDY. Relatively high TUNEL levels, reduction of proliferating cell nuclear antigen and cyclin-dependent kinase 4 expression, and fragmentation of poly (ADP-ribose) polymerase were also elucidated. These results suggest that commercial oral wet wipes could exert cytotoxic influences on oral tissue, although there are anti-bacterial effects, and careful attention is required, especially for infants and toddlers.

Ischemic Preconditioning and Its Relation to Glycogen Depletion (허혈성 전처치와 당원 결핍과의 관계)

  • 장대영;김대중;원경준;조대윤;손동섭;양기민;라봉진;김호덕
    • Journal of Chest Surgery
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    • v.33 no.7
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    • pp.531-540
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    • 2000
  • Baclgrpimd; Recent studies have suggested that the cardioprotective effect of ischemic preconditioning(IP) is closely related to glycogen depletion and attenuation of intracellular acidosis. In the present study, the authors tested this hypothesis by perfusion isolated rabbit hearts with glucose(G) is closely related to glycogen depletion and attenuation of intracellular acidosis. In the present study, the authors tested this hypothesis by perfusion isolated rabbit hearts with glucose(G)-free perfusate. Material and Method; Hearts isolated from New Zealand white rabbits(1.5~2.0 kg body weight) were perfused with Tyrode solution by Langendorff technique. After stabilization of baseline hemodynamics, the hearts were subjected to 45 min global ischemia followed by 120 min reperfusion with IP(IP group, n=13) or without IP(ischemic control group, n=10). IP was induced by single episode of 5 min global ischemia and 10 min reperfusion. In the G-free preconditioned group(n=12), G depletion was induced by perfusionwith G-free Tyrode solution for 5 min and then perfused with G-containing Tyrode solution for 10 min; and 45 min ischemia and 120 min reperfusion. Left ventricular functionincluding developed pressure(LVDP), dP/dt, heart rate, left ventricular end-distolic pressure(LVEDP) and coronary flow (CF) were measured. Myocardial cytosolic and membrane PKC activities were measured by 32P-${\gamma}$-ATP incorporation into PKC-specific peptide and PKC isozymes were analyzed by Western blot with monoclonal antibodies. Infarct size was determined by staining with TTC(tetrazolium salt) and planimetry. Data were analyzed by one-way analysis of variance (ANOVA) and Turkey's post-hoc test. Result ; In comparison with the ischemic control group, IP significantly enhanced functional recovery of the left ventricle; in contrast, functional significantly enhanced functional recovery of the left ventricle; in contrast, functional recovery were not significantly different between the G-free preconditioned and the ischemic control groups. However, the infarct size was significantly reduced by IP or G-free preconditioning(39$\pm$2.7% in the ischemic control, 19$\pm$1.2% in the IP, and 15$\pm$3.9% in the G-free preconditioned, p<0.05). Membrane PKC activities were increased significantly after IP (119%), IP and 45 min ischemia(145%), G-free [recpmdotopmomg (150%), and G-free preconditioning and 45 min ischemia(127%); expression of membrane PKC isozymes, $\alpha$ and $\varepsilon$, tended to be increased after IP or G-free preconditioning. Conclusion; These results suggest that in isolated Langendorff-perfused rabbit heart model, G-free preconditioning (induced by single episode of 5 min G depletion and 10 min repletion) colud not improve post-ischemic contractile dysfunction(after 45-minute global ischemia); however, it has an infarct size-limiting effect.

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A Destructive Method in the Connection of the Algorithm and Design in the Digital media - Centered on the Rapid Prototyping Systems of Product Design - (디지털미디어 환경(環境)에서 디자인 특성(特性)에 관한 연구(硏究) - 실내제품(室內製品) 디자인을 중심으로 -)

  • Kim Seok-Hwa
    • Journal of Science of Art and Design
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    • v.5
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    • pp.87-129
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    • 2003
  • The purpose of this thesis is to propose a new concept of design of the 21st century, on the basis of the study on the general signification of the structures and the signs of industrial product design, by examining the difference between modern and post-modern design, which is expected to lead the users to different design practice and interpretation of it. The starting point of this study is the different styles and patterns of 'Gestalt' in the post-modern design of the late 20th century from modern design - the factor of determination in industrial product design. That is to say, unlike functional and rational styles of modern product design, the late 20th century is based upon the pluralism characterized by complexity, synthetic and decorativeness. So far, most of the previous studies on design seem to have excluded visual aspects and usability, focused only on effective communication of design phenomena. These partial studies on design, blinded by phenomenal aspects, have resulted in failure to discover a principle of fundamental system. However, design varies according to the times; and the transformation of design is reflected in Design Pragnanz to constitute a new text of design. Therefore, it can be argued that Design Pragnanz serves as an essential factor under influence of the significance of text. In this thesis, therefore, I delve into analysis of the 20th century product design, in the light of Gestalt theory and Design Pragnanz, which have been functioning as the principle of the past design. For this study, I attempted to discover the fundamental elements in modern and post-modern designs, and to examine the formal structure of product design, the users' aesthetic preference and its semantics, from the integrative viewpoint. Also, with reference to history and theory of design my emphasis is more on fundamental visual phenomena than on structural analysis or process of visualization in product design, in order to examine the formal properties of modern and post-modern designs. Firstly, In Chapter 1, 'Issues and Background of the Study', I investigated the Gestalt theory and Design Pragnanz, on the premise of formal distinction between modern and post-modern designs. These theories are founded upon the discussion on visual perception of Gestalt in Germany in 1910's, in pursuit of the principle of perception centered around visual perception of human beings. In Chapter 2, I dealt with functionalism of modern design, as an advance preparation for the further study on the product design of the late 20th century. First of all, in Chapter 2-1, I examined the tendency of modern design focused on functionalism, which can be exemplified by the famous statement 'Form follows function'. Excluding all unessential elements in design - for example, decoration, this tendency has attained the position of the international style based on the spirit of Bauhause - universality and regularity - in search of geometric order, standardization and rationalization. In Chapter 2-2, I investigated the anthropological viewpoint that modern design started representing culture in a symbolic way including overall aspects of the society - politics, economics and ethics, and its criticism on functionalist design that aesthetic value is missing in exchange of excessive simplicity in style. Moreover, I examined the pluralist phenomena in post-modern design such as kitsch, eclecticism, reactionism, hi-tech and digital design, breaking away from functionalist purism of modern design. In Chapter 3, I analyzed Gestalt Pragnanz in design in a practical way, against the background of design trends. To begin with, I selected mass product design among those for the 20th century products as a target of analysis, highlighting representative styles in each category of the products. For this analysis, I adopted the theory of J. M Lehnhardt, who gradated in percentage the aesthetic and semantic levels of Pragnantz in design expression, and that of J. K. Grutter, who expressed it in a formula of M = O : C. I also employed eight units of dichotomies, according to the G. D. Birkhoff's aesthetic criteria, for the purpose of scientific classification of the degree of order and complexity in design; and I analyzed phenomenal aspects of design form represented in each unit. For Chapter 4, I executed a questionnaire about semiological phenomena of Design Pragnanz with 28 units of antonymous adjectives, based upon the research in the previous chapter. Then, I analyzed the process of signification of Design Pragnanz, founded on this research. Furthermore, the interpretation of the analysis served as an explanation to preference, through systematic analysis of Gestalt and Design Pragnanz in product design of the late 20th century. In Chapter 5, I determined the position of Design Pragnanz by integrating the analyses of Gestalt and Pragnanz in modern and post-modern designs In this process, 1 revealed the difference of each Design Pragnanz in formal respect, in order to suggest a vision of the future as a result, which will provide systemic and structural stimulation to current design.

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