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61.
High-efficiency transduction of human lymphoid progenitor cells and expression in differentiated T cells. 总被引:3,自引:2,他引:1 下载免费PDF全文
D S An Y Koyanagi J Q Zhao R Akkina G Bristol N Yamamoto J A Zack I S Chen 《Journal of virology》1997,71(2):1397-1404
Gene therapy strategies for humans have been limited by low transduction efficiencies and poor expression of retroviral vectors in differentiated progeny cells carrying the transduced vector. Here we describe a strategy utilizing a cell surface reporter gene, murine thy-1.2, selectable by fluorescence-activated cell sorting (FACS), to achieve higher gene marking efficiencies. Human CD34-positive cells were transduced by a murine retroviral vector bearing the thy-1.2 marker and pseudotyped with vesicular stomatitis virus G protein, followed by FACS to enrich for CD34-positive cells that express Thy-1.2 on the cell surface. Gene marking and expression after differentiation into thymocytes were assessed in a SCID-hu Thy/Liv mouse model for human lymphoid progenitor cell gene therapy. We found that virtually all of the differentiated T-cell progeny were marked with vector sequences. It is of particular importance that reconstitution with the selected cells resulted in expression of Thy-1.2 in up to 71% of donor-derived thymocytes. It is of note that the donor-derived thymocytes that did not express Thy-1.2 still harbored vector thy-1.2 sequences, suggesting repression of transgene expression in some cells during progenitor cell differentiation into thymocytes. These studies provide a proof of concept for efficient expression of transgenes through T-lymphoid differentiation and a potential basis for utilizing similar strategies in human gene therapy clinical trials. 相似文献
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本文通过同化枝腋芽丛生方式建立了盐生植物盐节木的快速繁殖体系。以在MS附加200 mmol·L-1 Na Cl的培养基上无菌种子萌发、生长共计4~5个月、高约3~4 cm的盐节木小苗为材料,从上截取0.5 cm长的同化枝小段为外植体,培养在MS附加不同浓度的6-BA与NAA的培养基中。结果表明:同化枝腋芽诱导与增殖最佳培养基均为MS+0.05 mg·L-1 6-BA+1mg·L-1 NAA或MS+1 mg·L-1 6-BA,初代培养30 d后,腋芽诱导率达97.5%,平均芽增殖系数为2.50;继代增殖培养时,将同化枝切成1 cm长的小段芽增殖系数提高到4.92;一次性芽苗伸长与生根培养基为MS+0.05 mg·L-1 IBA,生根率达99%;小苗经炼苗后移栽到沙子、营养土和蛭石体积比为1:1:1的基质中,成活率可达91%。 相似文献
63.
Positive moods are thought to restore self-control resources following depletion. However, it is not well understood whether this effect is due to affective valence (pleasantness), arousal (activation), or a combination of both. Across four studies, we set out to investigate the role of positive moods on cognitive and behavioral measures of self-regulation in an ego-depletion paradigm. In studies 1 and 2, we independently manipulated affective valence and arousal and assessed self-regulation with a Stroop task. Results did not suggest a restorative effect of either on cognitive resources. In study 3, we employed both behavioral (the ‘handgrip task’) and cognitive (Stroop) assessments of self-regulation. Again, no significant effect of mood was observed on the Stroop task. Additionally, participants did not persist significantly longer on the handgrip task following a positive mood induction. Finally, in study 4, high vs. low states of arousal were manipulated and self-regulation was assessed via pre- and post-manipulation Stroop performance. In study 4, Stroop performance improved slightly more across time points for those in the high arousal condition than for those in the low arousal condition. Therefore, across four studies, we failed to find a consistent pattern of results suggesting that positive moods restore cognitive resources. 相似文献
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Michael C. P. Livingston Sydney Bass Arnold W. Emery Thomas A. Thomson Gerrard A. Vaughan Wilfred T. Y. Wong Ronald N. Youngash Phillip S. Zack 《CMAJ》1973,109(10):1013-1016
This paper describes part of an education experiment at the University of British Columbia at Vancouver.Six final-year medical students spent approximately 12 weeks in a community. Their time was divided between the hospital and various doctors'' offices. They answered a simple questionnaire to describe their experiences and commented favourably upon the opportunities for direct patient contact, learning basic skills, informal teaching by both family physicians and consultants, and the variety of work available.They had the opportunity to follow up the progress of the patient and learn the natural history of common illnesses. They achieved their basic objectives. We conclude from their reports and informal conversation that the experiment was successful and recommend other institutions to try similar programs. 相似文献
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Effect of repeat copy number on variable-number tandem repeat mutations in Escherichia coli O157:H7 下载免费PDF全文
Variable-number tandem repeat (VNTR) loci have shown a remarkable ability to discriminate among isolates of the recently emerged clonal pathogen Escherichia coli O157:H7, making them a very useful molecular epidemiological tool. However, little is known about the rates at which these sequences mutate, the factors that affect mutation rates, or the mechanisms by which mutations occur at these loci. Here, we measure mutation rates for 28 VNTR loci and investigate the effects of repeat copy number and mismatch repair on mutation rate using in vitro-generated populations for 10 E. coli O157:H7 strains. We find single-locus rates as high as 7.0 x 10(-4) mutations/generation and a combined 28-locus rate of 6.4 x 10(-4) mutations/generation. We observed single- and multirepeat mutations that were consistent with a slipped-strand mispairing mutation model, as well as a smaller number of large repeat copy number mutations that were consistent with recombination-mediated events. Repeat copy number within an array was strongly correlated with mutation rate both at the most mutable locus, O157-10 (r2= 0.565, P = 0.0196), and across all mutating loci. The combined locus model was significant whether locus O157-10 was included (r2= 0.833, P < 0.0001) or excluded (r2= 0.452, P < 0.0001) from the analysis. Deficient mismatch repair did not affect mutation rate at any of the 28 VNTRs with repeat unit sizes of >5 bp, although a poly(G) homomeric tract was destabilized in the mutS strain. Finally, we describe a general model for VNTR mutations that encompasses insertions and deletions, single- and multiple-repeat mutations, and their relative frequencies based upon our empirical mutation rate data. 相似文献