Characterisation of human bronchial epithelial response to injury and identification of potential resident progenitor cells in vitro

As the first line of defence against external stimuli, the airway epithelium undergoes frequent injury during an adult life. This is countered by repair mechanisms that ensure the integrity of the epithelium. It has been established that there are resident stem/progenitor cells localized within specific niches throughout the respiratory tract. Stem/progenitor cells are activated according to the severity of the insult and are thought to be responsible for repairing the airway epithelium. It has been difficult to isolate those stem cells. In this study, an in vitro human bronchial epithelial model was adapted and characterised. In culture, a pseudostratified epithelium was observed, with basal, ciliated, and secretory cells. Mucus production was also seen in this model. A scrape-wound of the model was employed to study the responses of the airway epithelium to injury. It was observed that migration and then proliferation, of CD44 expressing basal cells, are the first events that take place after scrape-wounding. Up-regulation of CD44 was also observed at the edge of the wounds early post-wounding. This suggested a key role for CD44-expressing basal cells in migration and proliferation after wounding, also suggesting this population may contain a progenitor cell population. Investigation of the secretory profile of the airway epithelium post-wounding revealed an increase in a number of cytokines and growth factors. In particular, IL-6, IL-8, ENA-78, and RANTES were all elevated compared with unwounded cultures. A side population (SP) was identified in differentiated and undifferentiated human bronchial epithelial cells in at least some cultures accounting for 0.1-1.15% of cells present. In summary, the epithelium is important in airway wound repair with basal cells appearing to contain the progenitor population in this human bronchial epithelial cell (HBEC) model. Moreover, SP studies suggested the presence of SP in at least some cultures, which might contribute to airway regeneration. The secretory profile of the airway epithelium post-wounding indicates up-regulation of specific cytokines, which may be important in the pathogenesis of lung diseases such as asthma and COPD. This model should prove useful to assess wound repair pathways and may be of use in the future for proof of concept studies on novel therapeutic agents

Characterisation of human bronchial epithelial response to injury and identification of potential resident progenitor cells in vitro

As the first line of defence against external stimuli, the airway epithelium undergoes frequent injury during an adult life. This is countered by repair mechanisms that ensure the integrity of the epithelium. It has been established that there are resident stem/progenitor cells localized within specific niches throughout the respiratory tract. Stem/progenitor cells are activated according to the severity of the insult and are thought to be responsible for repairing the airway epithelium. It has been difficult to isolate those stem cells. In this study, an in vitro human bronchial epithelial model was adapted and characterised. In culture, a pseudostratified epithelium was observed, with basal, ciliated, and secretory cells. Mucus production was also seen in this model. A scrape-wound of the model was employed to study the responses of the airway epithelium to injury. It was observed that migration and then proliferation, of CD44 expressing basal cells, are the first events that take place after scrape-wounding. Up-regulation of CD44 was also observed at the edge of the wounds early post-wounding. This suggested a key role for CD44-expressing basal cells in migration and proliferation after wounding, also suggesting this population may contain a progenitor cell population. Investigation of the secretory profile of the airway epithelium post-wounding revealed an increase in a number of cytokines and growth factors. In particular, IL-6, IL-8, ENA-78, and RANTES were all elevated compared with unwounded cultures. A side population (SP) was identified in differentiated and undifferentiated human bronchial epithelial cells in at least some cultures accounting for 0.1-1.15% of cells present. In summary, the epithelium is important in airway wound repair with basal cells appearing to contain the progenitor population in this human bronchial epithelial cell (HBEC) model. Moreover, SP studies suggested the presence of SP in at least some cultures, which might contribute to airway regeneration. The secretory profile of the airway epithelium post-wounding indicates up-regulation of specific cytokines, which may be important in the pathogenesis of lung diseases such as asthma and COPD. This model should prove useful to assess wound repair pathways and may be of use in the future for proof of concept studies on novel therapeutic agents

Ancient and modern genomics of the Ohlone Indigenous population of California

Traditional knowledge, along with archaeological and linguistic evidence, documents that California supports cultural and linguistically diverse Indigenous populations. Studies that have included ancient genomes in this region, however, have focused primarily on broad-scale migration history of the North American continent, with relatively little attention to local population dynamics. Here, in a partnership involving researchers and the Muwekma Ohlone tribe, we analyze genomic data from ancient and present-day individuals from the San Francisco Bay Area in California: 12 ancient individuals dated to 1905 to 1826 and 601 to 184 calibrated years before the present (cal BP) from two archaeological sites and eight present-day members of the Muwekma Ohlone tribe, whose ancestral lands include these two sites. We find that when compared to other ancient and modern individuals throughout the Americas, the 12 ancient individuals from the San Francisco Bay Area cluster with ancient individuals from Southern California. At a finer scale of analysis, we find that the 12 ancient individuals from the San Francisco Bay Area have distinct ancestry from the other groups and that this ancestry has a component of continuity over time with the eight present-day Muwekma Ohlone individuals. These results add to our understanding of Indigenous population history in the San Francisco Bay Area, in California, and in western North America more broadly

A comparison of proteomic, genomic, and osteological methods of archaeological sex estimation

Sex estimation of skeletons is fundamental to many archaeological studies. Currently, three approaches are available to estimate sex–osteology, genomics, or proteomics, but little is known about the relative reliability of these methods in applied settings. We present matching osteological, shotgun-genomic, and proteomic data to estimate the sex of 55 individuals, each with an independent radiocarbon date between 2,440 and 100 cal BP, from two ancestral Ohlone sites in Central California. Sex estimation was possible in 100% of this burial sample using proteomics, in 91% using genomics, and in 51% using osteology. Agreement between the methods was high, however conflicts did occur. Genomic sex estimates were 100% consistent with proteomic and osteological estimates when DNA reads were above 100,000 total sequences. However, more than half the samples had DNA read numbers below this threshold, producing high rates of conflict with osteological and proteomic data where nine out of twenty conditional DNA sex estimates conflicted with proteomics. While the DNA signal decreased by an order of magnitude in the older burial samples, there was no decrease in proteomic signal. We conclude that proteomics provides an important complement to osteological and shotgun-genomic sex estimation

Defining the molecular role of gp91phox in the immune manifestation of acute allergic asthma using a preclinical murine model

<p>Abstract</p> <p>Objective</p> <p>The phenomena manifested during inflammation require interplay between circulating effector cells, local resident cells, soluble mediators and genetic host factors to establish, develop and maintain itself. Of the molecues involed in the initiation and perpetuation of acute allergic inflammation in asthma, the involvement of effector cells in redox reactions for producing O₂^-(superoxide anion) through the mediation of NADPH oxidase is a critical step. Prior data suggest that reactive oxygen species (ROS) produced by NADPH oxidase homologues in non-phagocytic cells play an important role in the regulation of signal transduction, while macrophages use a membrane-associated NADPH oxidase to generate an array of oxidizing intermediates which inactivate MMPs on or near them.</p> <p>Materials and Methods and Treatment</p> <p>To clarify the role of gp91phox subunit of NADPH oxidase in the development and progression of an acute allergic asthma phenotype, we induced allergen dependent inflammation in a gp91<it>^phox</it>-/- single knockout and a gp91phox-/-MMP-12-/- double knockout mouse models.</p> <p>Results</p> <p>In the knockout mice, both inflammation and airway hyperreactivity were more extensive than in wildtype mice post-OVA. Although OVA-specific IgE in plasma were comparable in wildtype and knockout mice, enhanced inflammatory cell recruitment from circulation and cytokine release in lung and BALf, accompanied by higher airway resistance as well as Penh in response to methacholine, indicate a regulatory role for NADPH oxidase in development of allergic asthma. While T cell mediated functions like Th2 cytokine secretion, and proliferation to OVA were upregulated synchronous with the overall robustness of the asthma phenotype, macrophage upregulation in functions such as proliferation, and mixed lymphocyte reaction indicate a regulatory role for gp91phox and an overall non-involvement or synergistic involvement of MMP12 in the response pathway (comparing data from gp91phox-/- and gp91phox-/-MMP-12-/- mice).</p

PI3K and ERK-Induced Rac1 Activation Mediates Hypoxia-Induced HIF-1α Expression in MCF-7 Breast Cancer Cells

Hypoxia-inducible factor 1 (HIF-1α) expression induced by hypoxia plays a critical role in promoting tumor angiogenesis and metastasis. However, the molecular mechanisms underlying the induction of HIF-1α in tumor cells remain unknown.In this study, we reported that hypoxia could induce HIF-1α and VEGF expression accompanied by Rac1 activation in MCF-7 breast cancer cells. Blockade of Rac1 activation with ectopic expression of an inactive mutant form of Rac1 (T17N) or Rac1 siRNA downregulated hypoxia-induced HIF-1α and VEGF expression. Furthermore, Hypoxia increased PI3K and ERK signaling activity. Both PI3K inhibitor LY294002 and ERK inhibitor U0126 suppressed hypoxia-induced Rac1 activation as well as HIF-1α expression. Moreover, hypoxia treatment resulted in a remarkable production of reactive oxygen species (ROS). N-acetyl-L-cysteine, a scavenger of ROS, inhibited hypoxia-induced ROS generation, PI3K, ERK and Rac1 activation as well as HIF-1α expression.Taken together, our study demonstrated that hypoxia-induced HIF-1α expression involves a cascade of signaling events including ROS generation, activation of PI3K and ERK signaling, and subsequent activation of Rac1

Comparative effectiveness and safety of non-vitamin K antagonists for atrial fibrillation in clinical practice: GLORIA-AF Registry

Background and purpose: Prospectively collected data comparing the safety and effectiveness of individual non-vitamin K antagonists (NOACs) are lacking. Our objective was to directly compare the effectiveness and safety of NOACs in patients with newly diagnosed atrial fibrillation (AF). Methods: In GLORIA-AF, a large, prospective, global registry program, consecutive patients with newly diagnosed AF were followed for 3&nbsp;years. The comparative analyses for (1) dabigatran vs rivaroxaban or apixaban and (2) rivaroxaban vs apixaban were performed on propensity score (PS)-matched patient sets. Proportional hazards regression was used to estimate hazard ratios (HRs) for outcomes of interest. Results: The GLORIA-AF Phase III registry enrolled 21,300 patients between January 2014 and December 2016. Of these, 3839 were prescribed dabigatran, 4015 rivaroxaban and 4505 apixaban, with median ages of 71.0, 71.0, and 73.0&nbsp;years, respectively. In the PS-matched set, the adjusted HRs and 95% confidence intervals (CIs) for dabigatran vs rivaroxaban were, for stroke: 1.27 (0.79–2.03), major bleeding 0.59 (0.40–0.88), myocardial infarction 0.68 (0.40–1.16), and all-cause death 0.86 (0.67–1.10). For the comparison of dabigatran vs apixaban, in the PS-matched set, the adjusted HRs were, for stroke 1.16 (0.76–1.78), myocardial infarction 0.84 (0.48–1.46), major bleeding 0.98 (0.63–1.52) and all-cause death 1.01 (0.79–1.29). For the comparison of rivaroxaban vs apixaban, in the PS-matched set, the adjusted HRs were, for stroke 0.78 (0.52–1.19), myocardial infarction 0.96 (0.63–1.45), major bleeding 1.54 (1.14–2.08), and all-cause death 0.97 (0.80–1.19). Conclusions: Patients treated with dabigatran had a 41% lower risk of major bleeding compared with rivaroxaban, but similar risks of stroke, MI, and death. Relative to apixaban, patients treated with dabigatran had similar risks of stroke, major bleeding, MI, and death. Rivaroxaban relative to apixaban had increased risk for major bleeding, but similar risks for stroke, MI, and death. Registration: URL: https://www.clinicaltrials.gov. Unique identifiers: NCT01468701, NCT01671007. Date of registration: September 2013