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Clara cell secretory protein deficiency alters clara cell secretory apparatus and the protein composition of airway lining fluid 总被引:2,自引:0,他引:2
Stripp BR Reynolds SD Boe IM Lund J Power JH Coppens JT Wong V Reynolds PR Plopper CG 《American journal of respiratory cell and molecular biology》2002,27(2):170-178
Clara cells represent the predominant secretory cell within distal conducting airways of mammals and exhibit functional alterations with chronic lung disease. We previously demonstrated that Clara cell secretory protein (CCSP) deficiency results in enhanced susceptibility to environmental agents. The present study was undertaken to define changes in Clara cell secretory function associated with CCSP deficiency in knockout mice. Comparative morphometry of Clara cell ultrastructure revealed dramatic alterations in secretory apparatus between wild-type (WT) and CCSP knockout (CCSP-/-) mice. Secretory granules, which occupy greater than 2% of Clara cell cytoplasmic volume in WT mice, were completely absent among Clara cells of CCSP-/- mice. Moreover, Clara cells of CCSP-/- mice exhibited a > 95% reduction in rough endoplasmic reticulum and alterations to Golgi apparatus, relative to WT controls. Ultrastructural perturbations to Clara cells were associated with altered protein composition of airway lining fluid as revealed by two-dimensional gel analysis of bronchoalveolar lavage proteins, but were not associated with altered abundance or secretion of CC26, another Clara cell secretory protein. We conclude that CCSP is required for the appearance of Clara cell secretory granules and that functional changes to Clara cells that result from CCSP deficiency lead to alterations in the composition of epithelial lining fluid. 相似文献
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In order to assess patients' preferences for different inhalation devices a questionnaire has been developed. The questionnaire consisted of items covering various aspects considered important for patients who regularly use inhalation devices. The questions were designed to be of a general character so any inhalation device could be evaluated. We evaluated the questionnaire in a multicenter, open crossover study comparing patients' opinions of MDI with and without spacer and of a dry powder inhaler, Turbuhaler. One hundred twenty-three patients with stable asthma confirmed by daily peak expiratory flow rate (PEF) measurements used for 2 weeks MDI only and MDI with spacer and for 2 weeks Turbuhaler. The order was randomized. The drugs were terbutaline inhaled via MDI and Turbuhaler and budesonide inhaled via MDI with spacer and turbuhaler. At the end of each evaluation period the patients answered a number of questions about their opinion by marking on a visual analogue scale. A factor analysis revealed that the different questions represent a few common factors important for evaluating opinion about inhalation devices. These factors are handling of the device, use, time to learn how to use the device properly, how confident the patients feel about the use of it, and taste/irritation. A standardized questionnaire of reduced size seems to be a suitable method for evaluating patient preference for different inhalation devices. 相似文献
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Birgitte Sandfeld-Paulsen Christina Demuth Birgitte H. Folkersen Torben R. Rasmussen Line B. Madsen Boe S. Sorensen 《Scandinavian journal of clinical and laboratory investigation》2016,76(3):243-248
Background Isolating sufficient material for molecular testing remains challenging in non-small cell lung cancer (NSCLC). The use of new ultra-microsamples (uMS) is proven sufficient for DNA and mRNA detection, but whether uMS are useful for quantifying mRNA expression is unknown. We investigated if uMS from lung cancer patients can be used to generate quantitative data on mRNA expression. Methods uMS were collected from primary tumors and lymph nodes from patients suspected of having lung cancer. mRNA was isolated, reverse-transcribed into cDNA and quantified with quantitative PCR assays for hepatocyte growth factor receptor (MET), hepatocyte growth factor (HGF), epidermal growth factor receptor (EGFR) and amphiregulin (AREG) mRNA. The fraction of tumor cells to normal cells was estimated in each sample. Results MET, HGF, EGFR, and AREG expression were evaluated in 90 samples (30 containing cancer cells and 60 without cancer cells). MET and EGFR expression were negligible in samples without cancer cells. In samples containing cancer cells, MET and EGFR could be quantified in 13 samples each. Adjustment for tumor-cell fraction made it possible to obtain a quantitative result for the tumor-cell mRNA expression of MET and EGFR. In contrast, AREG and HGF were expressed in samples without tumor cells. These samples were used to establish the AREG and HGF mRNA expression in normal cells. Seven out of 14?AR-positive and two out of eight HGF-positive samples with tumor cells were above a cut-off of the mean?+?2SD established in samples without tumor cells. Conclusion We demonstrate that uMS contain high-quality mRNA, and quantitative studies can be performed when the tumor-cell fraction is considered. 相似文献
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Targeted ultradeep next‐generation sequencing as a method for KIT D816V mutation analysis in mastocytosis
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Thomas Kristensen Sigurd Broesby‐Olsen Hanne Vestergaard Carsten Bindslev‐Jensen Michael Boe Mller 《European journal of haematology》2016,96(4):381-388
Next‐generation sequencing (NGS) is becoming increasingly used for diagnostic mutation analysis in myeloid neoplasms and may also represent a feasible technique in mastocytosis. However, detection of the KIT D816V mutation requires a highly sensitive method in most patients due to the typically low mutation levels. In this study, we established an NGS‐based KIT mutation analysis and analyzed the sensitivity of D816V detection using the Ion Torrent platform. Eighty‐two individual NGS analyses were included in the study. All samples were also analyzed using highly sensitive KIT D816V mutation‐specific qPCR. Measurements of the background level in D816V‐negative samples supported a cutoff for positivity of 0.2% in three different NGS panels. Clinical samples from patients with SM that tested positive using qPCR with a D816V allele burden >0.2% also tested positive using NGS. Samples that tested positive using qPCR with an allele burden <0.2% tested negative using NGS. We thereby demonstrate that caution should be taken when using the potentially very sensitive NGS technique for KIT D816V mutation analysis in mastocytosis, as many patients with SM have D816V mutation levels below the detection limit of NGS. A dedicated and highly sensitive KIT D816V mutation analysis therefore remains important in mastocytosis diagnostics. 相似文献
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S. R. McWhinney A. Tremblay S. G. Boe T. Bardouille 《Medical & biological engineering & computing》2018,56(2):201-210
Neurofeedback training teaches individuals to modulate brain activity by providing real-time feedback and can be used for brain–computer interface control. The present study aimed to optimize training by maximizing engagement through goal-oriented task design. Participants were shown either a visual display or a robot, where each was manipulated using motor imagery (MI)-related electroencephalography signals. Those with the robot were instructed to quickly navigate grid spaces, as the potential for goal-oriented design to strengthen learning was central to our investigation. Both groups were hypothesized to show increased magnitude of these signals across 10 sessions, with the greatest gains being seen in those navigating the robot due to increased engagement. Participants demonstrated the predicted increase in magnitude, with no differentiation between hemispheres. Participants navigating the robot showed stronger left-hand MI increases than those with the computer display. This is likely due to success being reliant on maintaining strong MI-related signals. While older participants showed stronger signals in early sessions, this trend later reversed, suggesting greater natural proficiency but reduced flexibility. These results demonstrate capacity for modulating neurofeedback using MI over a series of training sessions, using tasks of varied design. Importantly, the more goal-oriented robot control task resulted in greater improvements. 相似文献
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