• Title/Summary/Keyword: Culture Fusion

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Developmental Ability of Bovine Embryos Nuclear Transferred with Frozen-thawed or Cooled Donor Cells

  • Hong, S.B.;Uhm, S.J.;Lee, H.Y.;Park, C.Y.;Gupta, M.K.;Chung, B.H.;Chung, K.S.;Lee, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1242-1248
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    • 2005
  • This study was designed to investigate the in vitro developmental ability and apoptosis of bovine embryos nucleartransferred (NT) with frozen-thawed or cooled donor cells. Cultured adult bovine ear cells were used as donor cells after sub-culturing to confluence (CC), cooling to 4$^{\circ}C$ for 48 h, or freezing-thawing (FT). Apoptotic cells in blastocysts were evaluated for apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. Fusion, cleavage and blastocyst rates were 69.0 (167/242), 68.8 (115/167), and 29.9 (50/167) with CC cells, 70.4 (88/125), 69.3 (61/88), and 29.6 (26/88) with cooled cells and 66.1 (117/177), 70.1 (82/117), and 13.7 (16/117) with FT cells, respectively. Blastocyst rates of NT embryos derived from FT cells were significantly lower than those from CC or cooled cells (p<0.05). In addition, NT blastocysts produced by using FT cells showed significantly higher apoptosis rates (6.4${\pm}$4.0%) than those produced by CC (2.8${\pm}$1.7%) or cooled (2.3${\pm}$1.3%) cells. However, cooling of donor cells had no significant adverse effect on blastocyst rate as well as apoptosis rate. Therefore, our results suggest that cooled cells may be used as an alternative to freshly cultured confluent culture cells, as donor cells, for the production of Somatic nuclear cloned cattle.

Anti-Proliferative Effects of Dendrophthoe pentandra Methanol Extract on BCR/ABL-Positive and Imatinib-Resistant Leukemia Cell Lines

  • Zamani, Afiqah;Jusoh, Siti Asmaa Mat;Al-Jamal, Hamid Ali Nagi;Sul'ain, Mohd Dasuki;Johan, Muhammad Farid
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.11
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    • pp.4857-4861
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    • 2016
  • Background: Imatinib mesylate, a tyrosine kinase inhibitor specifically targeting the BCR/ABL fusion protein, induces hematological remission in patients with chronic myeloid leukemia (CML). However, the majority of CML patients treated with imatinib develop resistance with prolonged therapy. Dendrophthoe pentandra (L.) Miq. is a Malaysian mistletoe species that has been used as a traditional treatment for several ailments such as smallpox, ulcers, and cancers. Methods: We developed a resistant cell line (designated as K562R) by long-term co-culture of a BCR/ABL positive CML cell line, K562, with imatinib mesylate. We then investigated the anti-proliferative effects of D. pentandra methanol extract on parental K562 and resistant K562R cells. Trypan blue exclusion assays were performed to determine the IC50 concentration; apoptosis and cell cycle analysis were conducted by flow cytometry. Results: D. pentandra extract had greater anti-proliferative effects towards K562R ($IC50=192{\mu}g/mL$) compared to K562 ($500{\mu}g/mL$) cells. Upon treatment with D. pentandra extract at the IC50. concentration: K562 but not K562R demonstrated increase in apoptosis and cell cycle arrest in the G2/M phase. Conclusion: D. pentandra methanol extract exerts potent anti-proliferative effect on BCR/ABL positive K562 cells.

Expression and Characterization of Human N-Acetylglucosaminyltransferases and ${\alpha}$2,3-Sialyltransferase in Insect Cells for In Vitro Glycosylation of Recombinant Erythropoietin

  • Kim, Na-Young;Kim, Hyung-Gu;Kim, Yang-Hyun;Chung, In-Sik;Yang, Jai-Myung
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.383-391
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    • 2008
  • The glycans linked to the insect cell-derived glycoproteins are known to differ from those expressed in mammalian cells, partly because of the low level or lack of glycosyltransferase activities. GnT II, GnT IV, GnT V, and ST3Gal IV, which play important roles in the synthesis of tetraantennarytype complex glycan structures in mammalian cells, were overexpressed in Trichoplusia ni cells by using a baculovirus expression vector. The glycosyltransferases, expressed as a fusion form with the IgG-binding domain, were secreted into the culture media and purified using IgG sepharose resin. The enzyme assay, performed using a pyridylaminated-sugar chain as an acceptor, indicated that the purified glycosyltransferases retained their enzyme activities. Human erythropoietin expressed in T. ni cells (rhEPO) was subjected to in vitro glycosylation by using recombinant glycosyltransferases and was converted into complex-type glycan with terminal sialic acid. The presence of Nacetylglucosamine, galactose, and sialic acid on the rhEPO moiety was detected by a lectin blot analysis, and the addition of galactose and sialic acid to rhEPO was confirmed by autoradiography using $UDP-^{14}C-Gal\;and\;CMP-^{14}C-Sia$ as donors. The in vitro glycosylated rhEPO was injected into mice, and the number of reticulocytes among the ed blood cells was counted using FACS. A significant increase in the number of reticulocytes was not observed in the mice injected with in vitro glycosylated rhEPO as compared with those injected with rhEPO.

Recognition and Satisfaction fusion survey regarding health insurance coverage of dental scaling (치석제거 건강보험 급여화에 대한 일부 지역주민의 인식도 및 만족도 융합연구)

  • Ji, Min-Gyeong;Lee, Mi-Ra;Jung, Su-Jin
    • Journal of Convergence for Information Technology
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    • v.8 no.1
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    • pp.15-25
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    • 2018
  • This study aimed to investigate recognition of dental scaling and recognition of and satisfaction with health insurance coverage of dental scaling and make an efficient healthcare policy for dental health insurance; to do this, a survey was conducted in 389 residents in North Jeolla Province, drawing the following conclusion: First, the respondents were most likely to suggest that it was desirable to get dental scaling 'twice a year' and to start it 'in their twenties'. Second, those who were female and who were more concerned about periodontal health were more likely to recognize coverage of dental scaling. Third, those who were female and who were more concerned about periodontal health were also more satisfied with the coverage. It is therefore necessary to make PRs by using teaching media positively as well as by giving correct information with the objective of improving recognition of dental scaling on a continuous basis.

A Study on Standardization Plan to Foster Hanji Industry (한지산업 육성을 위한 표준화 방안 연구)

  • Jang, Se Gil
    • Journal of Agricultural Extension & Community Development
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    • v.21 no.3
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    • pp.121-154
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    • 2014
  • This study explores standardization plans concerning industrialization of Hanjii which is traditional culture resources of rural community. Hanji is very useful in many ways, and its usefulness is not confined to only paper. In other words, it comes to have architectural uses, agricultural uses, clothing uses, and medical uses through fusion and convergence. However, the expansion of Hanji market dose not lead to profits for related stake holders because its standardization is not nurtured yet. For standardization, this paper argues, first, a industrial definition of Hanji is needed in which Hanji's own characteristics is involved. Besides maintaining traditional characteristics, the definition and quality standards of Hanji needs to be established by considering a given condition of Hanji industy. Second, a satisfactory method to measure Hanji's own material characteristics(i.g., mulberry paper content and long-term sustainability), which affects Hanji's quality, needs to be developed and standardized. Third, a government standard is urgently needed in order to keep consumers from being confused when they select Hanji products. The standardization of Hanji products and technology needs to be established in both case of current Hanji market and newly developed products and technology.

Expression of Fungal Phytase on the Cell Surface of Saccharomyces cerevisiae

  • Mo, Ae-Young;Park, Seung-Moon;Kim, Yun-Sik;Yang, Moon-Sik;Kim, Dae-Hyuk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.576-581
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    • 2005
  • Phytase improves the bioavailability of phytate phosphorus in plant foods to humans and animals, and reduces the phosphorus pollution of animal waste. We have engineered the cell surface of the yeast. Saccharomyces cerevisiae, by anchoring active fungal phytase on its cell wall, in order to apply it as a dietary supplement containing bioconversional functions in animal foods and a whole cell bio-catalyst for the treatment of waste. The phytase gene (phyA) of Aspergillus niger with a signal peptide of rice amylase 1A (Ramy1A) was fused with the gene encoding the C-terminal half (320 amino acid residues from the C-terminus) of yeast ${\alpha}-agglutinin$, a protein which is involved in mating and is covalently anchored to the cell wall. The resulting fusion construct was introduced into S. cerevisiae and expressed under the control of the constitutive glyceraldehydes-3-phosphate dehydrogenase (GPD) promoter. Phytase plate assay revealed that the surface-engineered cell exhibited a catalytically active opaque zone which was restricted to the margin of the colony. Additionally, the phytase activity was detected in the cell fraction, but was not detected in the culture medium when it was grown in liquid. These results indicate that the phytase was successfully anchored to the cell surface of yeast and was displayed as its active form. The amount of recombinant phytase on the surface of yeast cells was estimated to be 16,000 molecules per cell.

Translation elongation factor-1A1 (eEF1A1) localizes to the spine by domain III

  • Cho, Sun-Jung;Lee, Hyun-Sook;Dutta, Samikshan;Seog, Dae-Hyun;Moon, Il-Soo
    • BMB Reports
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    • v.45 no.4
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    • pp.227-232
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    • 2012
  • In vertebrates, there are two variants of eukaryotic peptide elongation factor 1A (eEF1A; formerly eEF-$1{\alpha}$), eEF1A1 and eEF1A2, which have three well-conserved domains ($D_I$, $D_{II}$, and $D_{III}$). In neurons, eEF1A1 is the embryonic type, which is expressed during embryonic development as well as the first two postnatal weeks. In the present study, EGFP-tagged eEF1A1 truncates were expressed in cortical neurons isolated from rat embryo (E18-19). Live cell images of transfected neurons showed that $D_{III}$-containing EGFP-fusion proteins (EGFP-$D_{III}$, -$D_{II-III}$, -$D_{I-III}$) formed clusters that were confined within somatodendritic domains, while $D_{III}$-missing ones (EGFP-$D_I$, -$D_{II}$, -$D_{I-II}$) and control EGFP were homogeneously dispersed throughout the neuron including axons. In dendrites, EGFP-$D_{III}$ was targeted to the heads of spine- and filopodia-like protrusions, where it was colocalized with $SynGAP{\alpha}$, a postsynaptic marker. Our data indicate that $D_{III}$ of eEF1A1 mediates formation of clusters and localization to spines.

A Study on the Application of Computers to the Development of Humor Image Fashion Design (컴퓨터를 활용한 유머 이미지 패션디자인 개발에 관한 연구)

  • 우세희;최현숙
    • Journal of the Korean Society of Costume
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    • v.53 no.5
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    • pp.65-77
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    • 2003
  • Due to the rapid changes occurring in many aspects of contemporary society, the need for a means to actively combat widespread feelings of emptiness and alienation among the public, while satisfying its visual pleasure, is increasing. Thus the need for humorous elements which bring freedom to the human psyche is urgently requested. Of course, the field of fashion cannot be left out in this trend, and humor image design is a good example of this. Humor image in fashion endeavors to release the tension accumulated in the modern world, while trying to find a way to recover the original pureness of mankind. Another aspect currently important is computers. The creation of images in modern visual art relies a lot upon computers. Traditional visual processes such as painting, photography and video are now merged within digital technology. and are now quite symbiotic to each other. With the development of computers came computer art. which uses all applicable functions of a computer to create art. Any artistic action which uses a computer in any stage of its creation can be called computer art. The common factor in humor and computer art in modern fashion can be classified as follows : repetition, deformation and distortion. exaggeration and abridgement. juxtaposition. and Tromp l'oeil. This study has placed its objective on the fusion of humor image fashion and computer art, by manufacturing a work with humor and computers, two important aspects of modern culture. Expanding the field of fashion design while promoting creativity In fashion by finding a verging point between art and science is also necessary. I have designed and made five costumes using the above cited techniques in computer humor images, on a theoretical basis.

Bone Marrow-derived Side Population Cells are Capable of Functional Cardiomyogenic Differentiation

  • Yoon, Jihyun;Choi, Seung-Cheol;Park, Chi-Yeon;Choi, Ji-Hyun;Kim, Yang-In;Shim, Wan-Joo;Lim, Do-Sun
    • Molecules and Cells
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    • v.25 no.2
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    • pp.216-223
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    • 2008
  • It has been reported that bone marrow (BM)-side population (SP) cells, with hematopoietic stem cell activity, can transdifferentiate into cardiomyocytes and contribute to myocardial repair. However, this has been questioned by recent studies showing that hematopoietic stem cells (HSCs) adopt a hematopoietic cell lineage in the ischemic myocardium. The present study was designed to investigate whether BM-SP cells can in fact transdifferentiate into functional cardiomyocytes. Phenotypically, BM-SP cells were $19.59%{\pm}9.00\;CD14^+$, $8.22%{\pm}2.72\;CD34^+$, $92.93%{\pm}2.68\;CD44^+$, $91.86%{\pm}4.07\;CD45^+$, $28.48%{\pm}2.24\;c-kit^+$, $71.09%{\pm}3.67\;Sca-1^+$. Expression of endothelial cell markers (CD31, Flk-1, Tie-2 and VEGF-A) was higher in BM-SP cells than whole BM cells. After five days of co-culture with neonatal cardiomyocytes, $7.2%{\pm}1.2$ of the BM-SP cells expressed sarcomeric ${\alpha}$-actinin as measured by flow cytometry. Moreover, BM-SP cells co-cultured on neonatal cardiomyocytes fixed to inhibit cell fusion also expressed sarcomeric ${\alpha}$-actinin. The co-cultured BM-SP cells showed neonatal cardiomyocyte-like action potentials of relatively long duration and shallow resting membrane potential. They also generated calcium transients with amplitude and duration similar to those of neonatal cardiomyocytes. These results show that BM-SP cells are capable of functional cardiomyogenic differentiation when co-cultured with neonatal cardiomyocytes.

Plant Molecular Farming Using Oleosin Partitioning Technology in Oilseeds

  • Moloney, Maurice-M.
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.4
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    • pp.197-201
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    • 1997
  • Plant seed oil-bodies or oleosomes ate the repository of the neutral lipid stored in seeds. These organelles in many oilseeds may comprise half of the total cellular volume. Oleosomes are surrounded by a half-unit membrane of phospholipid into which are embedded proteins called oleosins. Oleosins are present at high density on the oil-body surface and after storage proteins comprise the most abundant proteins in oilseeds. Oleosins are specifically targeted and anchored to oil-bodies after co-translation on the ER. It has been shown that the amino-acid sequences responsible for this unique targeting reside primarily in the central hydrophobic tore of the oleosin polypeptide. In addition, a signal-like sequence is found near the junction of the hydrophobic domain and ann N-terminal hydrophilic / amphipathic domain. This "signal" which is uncleaved is also essential for correct targeting. Oil-bodies and their associated oleosins may be recovered by floatation centrifugation of aqueous seed extracts. This simple partitioning step results in a dramatic enrichment for oleosins in the oil-body fraction. In the light of these properties, we reasoned that it would be feasible to create fusion proteins on oil-bodies comprising oleosins and an additional valuable protein of pharmaceutical or industrial interest. It was further postulated that if these proteins were displayed on the outer surface of oil-bodies, it would be possible to release them from the purified oil-bodies using chemical or proteolytic cleavage. This could result in a simple means of recovering high-value protein from seeds at a significant (i.e. commercial) scale. This procedure has been successfully reduced to practice for a wide variety of proteins of therapeutic, industrial and food no. The utillity of the method will be discussed using a blood anticoagulant, hirudin, and industrial enzymes as key examples.

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