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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 22 (2000), S. 217-223 
    ISSN: 1573-0603
    Keywords: Flow cytometry ; Muscle cell ; Porcine ; Transfection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A detailed methodology is described for fluorescence-activated cell sorting (FACS) of porcine muscle cells that have been transfected to express green fluorescent protein (GFP). Cells are liberated from porcine skeletal muscle and primary cultures are transfected with DNA encoding GFP. Primary cultures are subjected to immunocytochemistry using a primary muscle-specific monoclonal antibody followed by incubation with a phycoerythrin-conjugated second antibody. Transfected myoblasts aresorted from fibroblasts using forward angle light scatter and ninety degree light scatter, phycoerythrin fluorescence, and GFP fluorescence. These procedures allow for isolation of genetically-engineered porcine muscle cells more rapidly than traditional clonal selection procedures. Consequently, FACS provides porcine myoblast populations that retain the majority of their replicative capacity and are not contaminated with non-myogenic cells.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Photosynthesis research 64 (2000), S. 15-26 
    ISSN: 1573-5079
    Keywords: ascorbate peroxidase ; glutathione reductase ; oxidative stress ; ozone tolerance ; superoxide dismutase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The current study confirmed earlier conclusions regarding differential ozone (O3) tolerances of two soybean cultivars, Essex and Forrest, and evaluated antioxidant enzyme activities of these two varieties based on their performance under environmentally relevant, elevated O3 conditions. The experiment was conducted in open-top chambers in the field during the 1994 and 1995 growing seasons. Exposure of plants to moderately high O3 levels (62.9 nl l−1 air, 2-year seasonal average) caused chlorophyll loss and increased membrane permeability when compared to control plants grown in charcoal filtered air (24.2 nl l−1 air). The other effects of O3 treatment were decrease in seed yield, loss of total sulfhydryl groups, reduction of soluble protein content, and increase in guaiacol peroxidase activity in leaves of both cultivars. The O3-induced increase in guaiacol peroxidase activity was much smaller in cv. Essex leaflets. Cv. Essex had less leaf oxidative damage and smaller reduction in seed yield than cv. Forrest under elevated O3 conditions. During ozonation, mature leaflets of the more O3 tolerant cv. Essex had higher levels of glutathione reductase (30%), ascorbate peroxidase (13%), and superoxide dismutase (45%) activity than did mature leaflets of cv. Forrest. Cu,Zn-superoxide dismutase, which represented 95% of total superoxide dismutase activity in the two cultivars, appeared to be increased by O3 exposure in the leaflets of O3 tolerant cv. Essex but not in those of cv. Forrest. Cytosolic ascorbate peroxidase activity was also higher in leaflets of cv. Essex than in cv. Forrest regardless of O3 level. Stromal ascorbate peroxidase and Mn-superoxide dismutase activity did not appear to be involved in the O3 tolerance of the two soybean cultivars.
    Type of Medium: Electronic Resource
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