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Background:Flagellated protozoan of the genus Leishmania is the causative agent of vector-borne parasitic diseases of leishmaniasis. Since the production of recombinant pharmaceutical proteins requires the cultivation of host cells in a serum-free medium, the elimination of FBS can improve the possibility of large-scale culture of Leishmania parasite. In the current study, we aimed at evaluating a new serum-free medium in Leishmania parasite culture for future live Leishmania vaccine purposes. Methods:Recombinant L. tarentolae secreting PpSP15-EGFP and wild type L. major were cultured in serum-free (CSFM) and serum-supplemented medium. The growth rate, protein expression, and infectivity of cultured parasites in both conditions was then evaluated and compared. Results:Diff-Quick staining and epi-fluorescence microscopy examination displayed the typical morphology of L. major and L. tarentolae-PpSP15-EGFP promastigote grown in CSFM medium. The amount of EGFP expression was similar in CSMF medium compared to M199 supplemented with 5% FBS in flow cytometry analysis of L. tarentolae-PpSP15-EGFP parasite. Also, a similar profile of PpSP15-EGFP proteins was recognized in Western blot analysis of L. tarentolae-PpSP15-EGFP cultured in CSMF and the serum-supplemented medium. Footpad swelling and parasite load measurements showed the ability of CSFM medium to support the L. major infectivity in BALB/C mice. Conclusion:This study demonstrated that CSFM can be a promising substitute for FBS supplemented medium in parasite culture for live vaccination purposes. Key Words: Growth rate, L. major, L. tarentolae, Serum-free medium, PpSP15-EGFP protein  相似文献   
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Combined effects of different light regimes, 6L:6D and 12L:12D (hours light : hours dark, L:D) and algal foods of Scenedesmus quadricauda (S) and Chlorella vulgaris (C) on ephippia production and fatty acid (FA) profile of Ceriodaphnia quadrangula were investigated. The highest resting egg production, density, and specific growth rate were obtained with 6L:6D+S; followed by 12L:12D+S treatment. The C16:0 varied from 16.2 to 22.16 mg/g dry weight (d.w.); C18:1n?9 from 11.03 to 14.94 mg/g d.w; C18:2n?6 from 9.79 to 20.24 mg/g d.w; and C18:3n?3 from 6.98 to 16.02 mg/g d.w. The amounts of C20:4n?6 (4.36 mg/g d.w) and C20:5n?3 (3.35 mg/g d.w) at 6L:6D+S were highest compared to other treatments. The maximum saturated fatty acids and monounsaturated fatty acids were obtained at 6L:6D+C while maximum polyunsaturated fatty acids observed at 6L:6D+S and 12L:12D+C. The resting egg production and FAs profile could be improved when the frequency of changes from light to dark were increased. Our findings also indicate a significant increase in the quantity of C18:3n?3 in C. quadrangula populations during resting egg formation, hence, implying that manipulation of light regime and algal food can improve performance of C. quadrangula as a live food in aquaculture industries, especially for larval feeding.  相似文献   
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Landscape Ecology - Many large carnivores depend on habitat patches outside protected areas, as well as safe corridors between them. However, corridor assessments typically ignore potential...  相似文献   
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Background: Pectic acid extracted from plants increases the secretion of prolactin (PRL) when injected intravenously into ewes or fed to rats. Fragments of ewe hypophysis and lactating rabbit mammary gland incubated in vitro in the presence of pectic acid secreted more PRL and caseins compared to the controls. However, it is not known whether pectic acid directly stimulates PRL secretion in pituitary or interference of factors from hypophysis is required for this process. Methods: GH3/B6 cells, a clonal strain of rat pituitary, were cultured and incubated with pectic acid (2.5-100 mug/mL). The integrity of cells was examined under pectic acid treatment microscopically. Controls or pectic acid treated cells were assayed for their ability to produce PRL. The PRL was assayed by Western-blotting and Radioimmunoassay. Results: pectic acid did not have any significant effect on the viability of cells. After being incubated with pectic acid, the cells started to become circular and protuberant shape. The maximum stimulation and PRL secretion occurred at 100 mug/mL concentration within 30 min of incubation with pectic acid. Conclusion: pectic acid could stimulate the release of PRL in GH3/B6 cells in the short-term incubation. This result suggested that pectic acid is a non-toxic agent that could directly stimulate PRL secretion in pituitary cells without any interference of hypophysis.  相似文献   
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