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951.
952.
Djamiatun K van der Ven AJ de Groot PG Faradz SM Hapsari D Dolmans WM Sebastian S Fijnheer R de Mast Q 《PLoS neglected tropical diseases》2012,6(5):e1628
Background
Thrombocytopenia, bleeding and plasma leakage are cardinal features of severe dengue. Endothelial cell activation with exocytosis of Weibel-Palade bodies (WPBs) may play an etiological role in this condition.Methods and Principal Findings
In a cohort of 73 Indonesian children with dengue hemorrhagic fever (DHF), of which 30 with dengue shock syndrome (DSS), we measured plasma levels of the WPB constituents von Willebrand factor antigen (VWF:Ag), VWF propeptide and osteoprotegerin (OPG), together with activity levels of the VWF-cleaving enzyme ADAMTS-13 and the amount of VWF in a platelet binding conformation (VWF activation factor). Compared with healthy controls (n = 17), children with DHF/DSS had significantly higher levels of VWF:Ag, VWF propeptide and OPG and decreased ADAMTS-13 activity. The VWF activation factor was also significantly higher in DHF/DSS and highest in children who died. There were significant differences in the kinetics of the various WPB constituents: VWF propeptide and OPG levels decreased toward discharge, while VWF:Ag levels were lower than expected at enrollment with plasma levels increasing toward discharge. Moreover, VWF propeptide levels correlated better with markers of disease severity (platelet count, liver enzymes, serum albumin and pleural effusion index) than corresponding VWF levels. Together, these findings suggest that there is consumption of VWF in DHF/DSS. In 4 out of 15 selected children with low ADAMTS-13 levels on admission, we found a remarkable reduction in the large and intermediate VWF multimers in the discharge blood samples, consistent with an acquired von Willebrand disease.Conclusion
These findings suggest that severe dengue is associated with exocytosis of WPBs with increased circulating levels of VWF:Ag, VWF propeptide and OPG. High circulating levels of VWF in its active conformation, together with low ADAMTS-13 activity levels, are likely to contribute to the thrombocytopenia and complications of dengue. During the convalescence phase, qualitative defects in VWF with loss of larger VWF multimers may develop. 相似文献953.
Alexandra L. Whittaker Kerry A. Lymn Gordon S. Howarth 《Journal of applied animal welfare science : JAAWS》2016,19(4):363-374
Although the metabolic cage is commonly used for housing nonhuman animals in the laboratory, it has been recognized as constituting a unique stressor. Such an environment would be expected to affect behavioral change in animals housed therein. However, few studies have specifically addressed the nature or magnitude of this change. The current study sought to characterize the behavioral time budget of rats in metabolic cage housing in comparison to that of individually housed animals in standard open-top cages. Rats in metabolic cages spent less time moving, manipulating enrichment, and carrying out rearing behaviors, and there was a corresponding shift toward inactivity. In an applied Social Interaction Test, behavioral scoring implied that metabolic cage housing had an anxiogenic effect. In conclusion, metabolic cage housing produces measurable effects on spontaneous and evoked behavior in rats in the laboratory. These behavioral changes may lead to a negative emotional state in these animals, which could have negative welfare consequences. Further research is needed to quantify the existence and magnitude of such an effect on rat well being. 相似文献
954.
Sarah E. Adams Andrew G. Purkiss Phillip P. Knowles Andrea Nans David C. Briggs Annabel Borg Christopher P. Earl Kerry M. Goodman Agata Nawrotek Aaron J. Borg Pauline B. McIntosh Francesca M. Houghton Svend Kjær Neil Q. McDonald 《Structure (London, England : 1993)》2021,29(7):694-708.e7
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956.
Lutz Müller David Blakey Kerry L. Dearfield Sheila Galloway Peggy Guzzie Makoto Hayashi Peter Kasper David Kirkland James T. MacGregor James M. Parry Leonard Schechtman Andrew Smith Noriho Tanaka David Tweats Hiroshi Yamasaki 《Mutation Research - Genetic Toxicology and Environmental Mutagenesis》2003,540(2):177
957.
We have developed a PCR-based short interfering RNA (siRNA) quantification method based on competition between siRNA and a homologous DNA primer for annealing to template DNA, avoiding the requirement for prior conversion of RNA to cDNA. Primers and probe were designed to amplify regions of the human papillomavirus E6 or enhanced green fluorescent protein genes. Having confirmed siRNA could not act as primer for amplicon generation, the lowest competing primer concentration yielding a linear relationship between template DNA amount (0.1–50 ng) and cycle of threshold (Ct) was determined (6.25 nM). Under these conditions addition of sequence-specific siRNA to the competitive quantitative PCR (cqPCR), resulted in a dose-dependent linear increase in Ct value. 2′-O-methyl ribose-modified siRNA retained an ability to inhibit template amplification in serum, unlike unmodified siRNAs that were susceptible to endonucleases. Mismatch-bearing or truncated siRNAs failed to inhibit template amplification confirming sequence specificity and an ability to discriminate between degraded and non-degraded siRNA sequences. Following delivery of E6 siRNA to C33-A cells using oligofectamine or His6 reducible polymers, siRNA uptake was quantified by cqPCR, revealing dose-dependent uptake. We anticipate that cqPCR will allow accurate determination of siRNA pharmacokinetics following in vivo delivery, greatly facilitating development of therapeutic siRNA delivery strategies. 相似文献
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