New Limits to the Drift of Fundamental Constants from Laboratory Measurements

We have remeasured the absolute $1S$-$2S$ transition frequency $\nu_{\rm {H}}$ in atomic hydrogen. A comparison with the result of the previous measurement performed in 1999 sets a limit of $(-29\pm 57)$ Hz for the drift of $\nu_{\rm {H}}$ with respect to the ground state hyperfine splitting $\nu_{{\rm {Cs}}}$ in $^{133}$Cs. Combining this result with the recently published optical transition frequency in $^{199}$Hg$^+$ against $\nu_{\rm {Cs}}$ and a microwave $^{87}$Rb and $^{133}$Cs clock comparison, we deduce separate limits on $\dot{\alpha}/\alpha = (-0.9\pm 2.9)\times 10^{-15}$ yr$^{-1}$ and the fractional time variation of the ratio of Rb and Cs nuclear magnetic moments $\mu_{\rm {Rb}}/\mu_{\rm {Cs}}$ equal to $(-0.5 \pm 1.7)\times 10^{-15}$ yr$^{-1}$. The latter provides information on the temporal behavior of the constant of strong interaction.Comment: 4 pages, 3 figures, LaTe

The role of doxorubicin in non-viral gene transfer in the lung

a b s t r a c t Proteasome inhibitors have been shown to increase adeno-associated virus (AAV)-mediated transduction in vitro and in vivo. To assess if proteasome inhibitors also increase lipid-mediated gene transfer with relevance to cystic fibrosis (CF), we first assessed the effects of doxorubicin and N-acetyl-L-leucinyl-L-leucinal-L-norleucinal in non-CF (A549) and CF (CFTE29o-) airway epithelial cell lines. CFTE29o-cells did not show a response to Dox or LLnL; however, gene transfer in A549 cells increased in a dose-related fashion (p < 0.05), up to approximately 20-fold respectively at the optimal dose (no treatment: 9.3 Â 10 4 AE 1.5 Â 10 3 , Dox: 1.6 Â 10 6 AE 2.6 Â 10 5 , LLnL: 1.9 Â 10 6 AE 3.2 Â 10 5 RLU/mg protein). As Dox is used clinically in cancer chemotherapy we next assessed the effect of this drug on non-viral lung gene transfer in vivo. CF knockout mice were injected intraperitoneally (IP) with Dox (25-100 mg/kg) immediately before nebulisation with plasmid DNA carrying a luciferase reporter gene under the control of a CMV promoter/ enhancer (pCIKLux) complexed to the cationic lipid GL67A. Dox also significantly (p < 0.05) increased expression of a plasmid regulated by an elongation factor 1a promoter (hCEFI) approximately 8-fold. Although administration of Dox before lung gene transfer may not be a clinically viable option, understanding how Dox increases lung gene expression may help to shed light on intracellular bottle-necks to gene transfer, and may help to identify other adjuncts that may be more appropriate for use in man

Airway branching morphogenesis in three dimensional culture

To access publisher full text version of this article. Please click on the hyperlink in Additional Links fieldBACKGROUND: Lungs develop from the fetal digestive tract where epithelium invades the vascular rich stroma in a process called branching morphogenesis. In organogenesis, endothelial cells have been shown to be important for morphogenesis and the maintenance of organ structure. The aim of this study was to recapitulate human lung morphogenesis in vitro by establishing a three dimensional (3D) co-culture model where lung epithelial cells were cultured in endothelial-rich stroma. METHODS: We used a human bronchial epithelial cell line (VA10) recently developed in our laboratory. This cell line cell line maintains a predominant basal cell phenotype, expressing p63 and other basal markers such as cytokeratin-5 and -14. Here, we cultured VA10 with human umbilical vein endothelial cells (HUVECs), to mimic the close interaction between these cell types during lung development. Morphogenesis and differentiation was monitored by phase contrast microscopy, immunostainings and confocal imaging. RESULTS: We found that in co-culture with endothelial cells, the VA10 cells generated bronchioalveolar like structures, suggesting that lung epithelial branching is facilitated by the presence of endothelial cells. The VA10 derived epithelial structures display various complex patterns of branching and show partial alveolar type-II differentiation with pro-Surfactant-C expression. The epithelial origin of the branching VA10 colonies was confirmed by immunostaining. These bronchioalveolar-like structures were polarized with respect to integrin expression at the cell-matrix interface. The endothelial-induced branching was mediated by soluble factors. Furthermore, fibroblast growth factor receptor-2 (FGFR-2) and sprouty-2 were expressed at the growing tips of the branching structures and the branching was inhibited by the FGFR-small molecule inhibitor SU5402. DISCUSSION: In this study we show that a human lung epithelial cell line can be induced by endothelial cells to form branching bronchioalveolar-like structures in 3-D culture. This novel model of human airway morphogenesis can be used to study critical events in human lung development and suggests a supportive role for the endothelium in promoting branching of airway epithelium

DNA demethylation-dependent enhancement of toll-like receptor-2 gene expression in cystic fibrosis epithelial cells involves SP1-activated transcription

<p>Abstract</p> <p>Background</p> <p>The clinical course of cystic fibrosis (CF) is characterized by recurrent pulmonary infections and chronic inflammation. We have recently shown that decreased methylation of the toll-like receptor-2 (TLR2) promoter leads to an apparent CF-related up-regulation of TLR2. This up-regulation could be responsible, in part, for the CF-associated enhanced proinflammatory responses to various bacterial products in epithelial cells. However, the molecular mechanisms underlying DNA hypomethylation-dependent enhancement of TLR2 expression in CF cells remain unknown.</p> <p>Results</p> <p>The present study indicates that there is a specific CpG region (CpG#18-20), adjacent to the SP1 binding site that is significantly hypomethylated in several CF epithelial cell lines. These CpGs encompass a minimal promoter region required for basal TLR2 expression, and suggests that CpG#18-20 methylation regulates TLR2 expression in epithelial cells. Furthermore, reporter gene analysis indicated that the SP1 binding site is involved in the methylation-dependent regulation of the TLR2 promoter. Inhibition of SP1 with mithramycin A decreased TLR2 expression in both CF and 5-azacytidine-treated non-CF epithelial cells. Moreover, even though SP1 binding was not affected by CpG methylation, SP1-dependent transcription was abolished by CpG methylation.</p> <p>Conclusion</p> <p>This report implicates SP1 as a critical component of DNA demethylation-dependent up-regulation of TLR2 expression in CF epithelial cells.</p

Project ASSIST: School Culture Typology

A list of MOspace items relating to Project ASSIST may be found at https://mospace.umsystem.edu/xmlui/handle/10355/3481/browse?value=Project+ASSIST&type=subjectFurther information may be found on the Middle Level Leadership Center web site at http://education.missouri.edu/orgs/mllc/3A_ast_overview.phpFiles attached below (in order) are "Directions for Using the Culture Typology Activity," "Culture Typology Activity," "Culture Typology Descriptions," worksheet Examples A-C, "Culture Typology Reflection Sheet," "A Collaborative Culture for School Improvement: Significance, Definition, and Measurement" (research summary), "Cultural Typology Row Explanation", and "Culture Typology Consensus Worksheet."This is one of three Project ASSIST School Culture Activities. The other two items may be found at https://mospace.umsystem.edu/xmlui/handle/10355/3558 and https://mospace.umsystem.edu/xmlui/handle/10355/3560The Culture Typology Activity was developed for use by teachers and principals as a method to help schools and school leaders (a) identify the general type of culture present in a school, (b) reflect upon the impact of that type of culture on student success, and (c) stimulate the discussion and the design of strategies to develop and maintain a more collaborative culture

Project ASSIST: School Culture Survey

A list of MOspace items relating to Project ASSIST may be found at https://mospace.umsystem.edu/xmlui/handle/10355/3481/browse?value=Project+ASSIST&type=subjectFurther information may be found on the Middle Level Leadership Center web site at http://education.missouri.edu/orgs/mllc/3A_ast_overview.phpFiles attached below (in order) are the "School Culture Survey," "School Culture Survey with Factor Descriptions and Item Examples," and "School Culture Survey Factors and Items."This is one of three Project ASSIST School Culture Activities. The other two items may be found at https://mospace.umsystem.edu/xmlui/handle/10355/3560 and https://mospace.umsystem.edu/xmlui/handle/10355/3559The School Culture Survey provides insight about the shared values/beliefs, the patterns of behavior, and the relationships in the school. Each factor measures a unique aspect of the school's collaborative culture

Divergent Inhibitor Susceptibility among Airway Lumen-Accessible Tryptic Proteases.

Tryptic serine proteases of bronchial epithelium regulate ion flux, barrier integrity, and allergic inflammation. Inhibition of some of these proteases is a strategy to improve mucociliary function in cystic fibrosis and asthmatic inflammation. Several inhibitors have been tested in pre-clinical animal models and humans. We hypothesized that these inhibitors inactivate a variety of airway protease targets, potentially with bystander effects. To establish relative potencies and modes of action, we compared inactivation of human prostasin, matriptase, airway trypsin-like protease (HAT), and β-tryptase by nafamostat, camostat, bis(5-amidino-2-benzimidazolyl)methane (BABIM), aprotinin, and benzamidine. Nafamostat achieved complete, nearly stoichiometric and very slowly reversible inhibition of matriptase and tryptase, but inhibited prostasin less potently and was weakest versus HAT. The IC50 of nafamostat's leaving group, 6-amidino-2-naphthol, was &gt;104-fold higher than that of nafamostat itself, consistent with suicide rather than product inhibition as mechanisms of prolonged inactivation. Stoichiometric release of 6-amidino-2-naphthol allowed highly sensitive fluorometric estimation of active-site concentration in preparations of matriptase and tryptase. Camostat inactivated all enzymes but was less potent overall and weakest towards matriptase, which, however was strongly inhibited by BABIM. Aprotinin exhibited nearly stoichiometric inhibition of prostasin and matriptase, but was much weaker towards HAT and was completely ineffective versus tryptase. Benzamidine was universally weak. Thus, each inhibitor profile was distinct. Nafamostat, camostat and aprotinin markedly reduced tryptic activity on the apical surface of cystic fibrosis airway epithelial monolayers, suggesting prostasin as the major source of such activity and supporting strategies targeting prostasin for inactivation