• Title/Summary/Keyword: Cell isolation

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Perceived Anxiety and Uncertainty in Hematopoietic Stem Cell Transplantation Recipients to and from Isolation Unit (격리병동 입.퇴실시 조혈모세포 이식 환자가 지각하는 불안과 불확실성)

  • Kim, Hye-Jo;Choi, Dong-Won;Park, Ho-Ran;Sohng, Kyeong-Yae
    • Journal of Korean Academy of Fundamentals of Nursing
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    • v.13 no.3
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    • pp.428-436
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    • 2006
  • Purpose: To investigate the levels of uncertainty and anxiety at admission and discharge to the isolation unit for patients undergoing hematopoietic stem cell transplantation (HSCT). Method: The data were collected from 60 patients who were admitted to the department of HSCT. Uncertainty and anxiety were assessed using the Mishel Uncertainty in Illness Scale (MUIS) and Anxiety Inventory (STAI). Collected data were analyzed using the SAS program. Results: The mean score for anxiety at admission was 2.20, and at discharge 2.10. The mean score for uncertainty at admission was 2.19, and at discharge 2.07. The anxiety at admission the group with physical discomfort was higher than that of group without physical discomfort. A positive relationship was found between anxiety and uncertainty at admission and at discharge. The major variables were expectation for cure and physical discomfort, explaining 25.87% of the anxiety at admission. The major variable was expectation for cure, explaining 20.94% of the uncertainty at admission. Conclusion: Front the above results, it can be concluded that support and consideration are required to reduce anxiety of the inpatient in single room.

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Isolation and Characterization of Trophoblast Stem Cells-like Cells Derived from Human Term Placenta

  • Na, Kyu-Hwan;Shin, Kyung-Seon;Choi, Jong-Ho;Cha, Dong-Hyun;Kim, Gi-Jin
    • Development and Reproduction
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    • v.14 no.3
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    • pp.155-162
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    • 2010
  • The trophectoderm is one of the earliest cell types to differentiate in the forming placenta. It is an important for the initial implantation and placentation during pregnancy. Trophoblast stem cells (TBSCs) develop from the blastocyst and are maintained by signals emanating from the inner cell mass. However, several limitations including rarity and difficulty in isolation of trophoblast stem cells derived from blastocyst still exist. To establish a model for trophoblast differentiation, we isolated TBSCs from human term placenta ($\geq$38 weeks) and characterized. Cell cycle was analyzed by measuring DNA content by FACS analysis and phenotype of TBSCs was characterized by RT-PCR and FACS analysis. TBSCs have expressed various markers such as self-renewal markers (Nanog, Sox2), three germ layer markers (hNF68, alpha-cardiac actin, hAFP), trophoblast specific markers (CDX-2, CK7, HLA-G), and TERT gene. In FACS analysis, TBSCs isolated from term placenta showed that the majority of cells expressed CD13, CD44, CD90, CD95, CD105, HLA-ABC, cytokeratin 7, and HLA-G. Testing for CD31, CD34, CD45, CD71, vimentin and HLA-DR were negative. TBSCs were shown to decrease the growth rate when cultured in conditioned medium without FGF4/heparin as well as the morphology was changed to a characteristic giant cell with a large cytoplasm and nucleus. In invasion assay, TBSCs isolated from term placenta showed invasion activities in in vivo using nude mice and in vitro Matrigel system. Taken together, these results support that an isolation potential of TBSCs from term placenta as well as a good source for understanding of the infertility mechanism.

Isolation and Characterization of Purple Non-Sulfur Bacteria, Afifella marina, Producing Large Amount of Carotenoids from Mangrove Microhabitats

  • Soon, Tan Kar;Al-Azad, Sujjat;Ransangan, Julian
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1034-1043
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    • 2014
  • This study determined the effect of light intensity and photoperiod on the dry cell weight and total amount of carotenoids in four isolates of purple non-sulfur bacteria obtained from shaded and exposed microhabitats of a mangrove ecosystem in Kota Kinabalu, Sabah, Malaysia. The initial isolation of the bacteria was carried out using synthetic 112 medium under anaerobic conditions (2.5 klx) at $30{\pm}2^{\circ}C$. On the basis of colony appearance, cell morphology, gram staining, motility test, and 16S rRNA gene sequencing analyses, all four bacteria were identified as Afifella marina. One of the bacterial isolates, designated as Af. marina strain ME, which was extracted from an exposed mud habitat within the mangrove ecosystem, showed the highest yield in dry cell weight ($4.32{\pm}0.03g/l$) as well as total carotenoids ($0.783{\pm}0.002mg/g$ dry cell weight). These values were significantly higher than those for dry cell weight ($3.77{\pm}0.02g/l$) and total carotenoid content ($0.706{\pm}0.008mg/g$) produced by the isolates from shaded habitats. Further analysis of the effect of 10 levels of light intensity on the growth characteristics of Af. marina strain ME showed that the optimum production of dry cell weight and total carotenoids was achieved at different light intensities and incubation periods. The bacterium produced the highest dry cell weight of 4.98 g/l at 3 klx in 72 h incubation, but the carotenoid production of 0.783 mg/g was achieved at 2.5 klx in 48 h incubation. Subsequent analysis of the effect of photoperiod on the production of dry cell weight and total carotenoids at optimum light intensities (3 and 2.5 klx, respectively) revealed that 18 and 24 h were the optimum photoperiods for the production of dry cell weight and total carotenoids, respectively. The unique growth characteristics of the Af. marina strain ME can be exploited for biotechnology applications.

In Vitro Culture of Endothelial Cell and Smooth Muscle Cell for Studying Vascular Diseases

  • Kim, Joo-Young
    • Journal of Yeungnam Medical Science
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    • v.27 no.2
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    • pp.91-97
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    • 2010
  • Endothelial cells play a key role in pathological processes such as cancer cell metastasis, atherosclerosis, and diabetic retinopathy. Vascular smooth muscle cells directly involve in the formation of atheroma in atherosclerosis. Some kinds of the endothelial cells are simply harvested from the umbilical veins, the tunica intima of aortic walls, the retina using various enzymes solutions. Those purely isolated cells provide a powerful tool in vitro studies of the endothelial cell related diseases. In this context, the cultured smooth muscle cells after the isolation from the tunica media of aortic walls are also used for elucidating the pathogenesis of atherosclerosis. Here, I briefly introduce articles that include the isolation of human umbilical vein endothelial cells(HUVEC), aortic endothelial and smooth muscle cells, retinal microvascular endothelial cells(RMEC), as well as the diseases' applications of these cells.

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Remotely Operated Decontamination Systems for Use in DFDF

  • Kim, Kiho;Park, Jangjin;Myungseung Yang
    • Proceedings of the Korean Radioactive Waste Society Conference
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    • 2003.11a
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    • pp.438-446
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    • 2003
  • This paper presents the development of the remotely operated decontamination systems for use in a highly radioactive zone of the DUPIC Fuel Development facility of the Irradiated Material Examination Facility at the Korea Atomic Energy Research Institute. The remotely operated decontamination systems were designed to completely eliminate human interaction with hazardous radioactive contaminants. These decontamination systems are mainly classified into three systems depending on the task environment - a fabrication equipment decontamination system, a hot-cell floor decontamination system, and an isolation room floor decontamination system. A decontamination system for contaminated fabrication equipment utilizes dry ice pellet blasting method to decontaminate contaminated surface of the equipment. The decontamination systems for the hot-cell floor and isolation room floor employ a vacuum cleaning method to decontaminate the contaminated floor and collect loose dry spent nuclear fuel debris and other radioactive waste placed on the floor. The human operator from the out-of-cell performs a series of decontamination tasks remotely by manipulating decontamination systems located in-cell via a handcontroller with the aid of vision feedback information. The environmental, functional and mechanical design considerations, control system and capabilities of the remotely operated decontamination systems at a high radioactive environment are also described.

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Isolation and culture of protoplasts from leaf tissue of Capsicum annnum var. accumnatum Fingerh and C. frutescens L. [Syn. C. minmum Roxb.] (Bird chilli)

  • Lee, Kue-Jae;Lee, Wang-Hyu
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2003.10b
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    • pp.20-20
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    • 2003
  • Isolation and culture of leaf protoplasts from two chilli cultivars (Capsicum annuum var. accumnatum and Bird chilli) were developed to enhance selection process in the somatic hybridization programmes. In order to isolate the protoplasts from leaves of these two chilli cultivars different incubation periods (3, 5 and 10 hours) were tested with combinations of enzyme mixtures containing cellulase and macerozyme. Leaves were incubated on three enzyme mixtures (2% cellulase +0.4% macerozyme, 1% cellulase +0.2% macerozyme and 0.5% cellulase +0.1% macerozyme in 13% mannitol) at 251oC in the dark. Three hours of incubation using 2% cellulase and 0.4% macerozyme was the best for the protoplast isolation of both chilli cultivars tested. The yield was 5${\times}$108protoplasts/m1/g leaf tissue in both chilli varieties. It was found that in the mixed nurse method using Nagata and Takebe (NT) medium supplemented with 1.0mg/12,4-D, NAA and BAP with 0.5M mannitol and 1.2% Sea Plaque agarose is the best medium for protoplast culture. Protoplasts of Capsicum annum var. accumnatum were alive for 14 days forming cell walls and initiating cell division.

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Use of Clostridium septicum Alpha Toxins for Isolation of Various Glycosylphosphatidylinositol-Deficient Cells

  • Shin Dong-Jun;Choy Hyon E.;Hong Yeongjin
    • Journal of Microbiology
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    • v.43 no.3
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    • pp.266-271
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    • 2005
  • In eukaryotic cells, various proteins are anchored to the plasma membrane through glycosylphosphatidylinositol (GPI). To study the biosynthetic pathways and modifications of GPI, various mutant cells have been isolated from the cells of Chinese hamster ovaries (CHO) supplemented with several exogenous genes involved in GPI biosynthesis using aerolysin, a toxin secreted from gram-negative bacterium Aeromonas hydrophila. Alpha toxin from Gram-positive bacterium Clostridium septicum is homologous to large lobes (LL) of aerolysin, binds GPI-anchored proteins and possesses a cell-destroying mechanism similar to aerolysin. Here, to determine whether alpha toxins can be used as an isolation tool of GPI-mutants, like aerolysin, CHO cells stably transfected with several exogenous genes involved in GPI biosynthesis were chemically mutagenized and cultured in a medium containing alpha toxins. We isolated six mutants highly resistant to alpha toxins and deficient in GPI biosynthesis. By genetic complementation, we determined that one mutant cell was defective of the second subunit of dolichol phosphate mannose synthase (DPM2) and other five cells were of a putative catalytic subunit of inositol acyltransferase (PIG-W). Therefore, C. septicum alpha toxins are a useful screening probe for the isolation of various GPI-mutant cells.

A Rate-Controlled Cell Multiplexer in ATM Networks (ATM망에서 흐름제어 셀 다중화기)

  • Kang, Koo-Hong;Park, Sang-Jo
    • Journal of KIISE:Information Networking
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    • v.28 no.3
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    • pp.417-425
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    • 2001
  • Since First-Come-First-Served discipline makes it difficult to satisfy the diverse quality of scrvicc(QoS) requirements for each c-oonection, traffic isolation has been proposed for guaranteeing the desired QoS through explicit bandwidth allocation to each virtual connection(VC) in A TM networks. For this purpose, we propose a rate-controlled cell multiplexer using a simple "periodic token-based regulator" per- Vc. The proposed multiplexer realizes VC isolation, while still operating work'||'&'||'not;conservingly. We first discuss its major properties, and then examine its features with respect to the characteristics of the output traffic streams and the traffic isolation of individual VC's, by simulation.n.

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Improved Procedure for Large-scale Isolation of Mitochondrial DNA from Mammalian Tissues

  • Hong, Sung-Soo;Lee, Chung-Choo
    • Animal cells and systems
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    • v.3 no.1
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    • pp.73-78
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    • 1999
  • Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

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