Ciliated Epithelial Cell Differentiation at Air-Liquid Interface Using Commercially Available Culture Media

The human nasal epithelium contains basal stem/progenitor cells that produce differentiated multiciliated and mucosecretory progeny. Basal epithelial cells can be expanded in cell culture and instructed to differentiate at an air-liquid interface using transwell membranes and differentiation media. For basal cell expansion, we have used 3T3-J2 co-culture in epithelial culture medium containing EGF, insulin, and a RHO-associated protein kinase (ROCK) inhibitor, Y-27632 (3T3 + Y). Here we describe our protocols for ciliated differentiation of these cultures at air-liquid interface and compare four commercially available differentiation media, across nine donor cell cultures (six healthy, two patients with chronic obstructive pulmonary disease (COPD), and one with primary ciliary dyskinesia (PCD)). Bright-field and immunofluorescence imaging suggested broad similarity between differentiation protocols. Subtle differences were seen in transepithelial electrical resistance (TEER), ciliary beat frequency, mucus production, and the extent to which basal cells are retained in differentiated cultures. Overall, the specific differentiation medium used in our air-liquid interface culture protocol was not a major determinant of ciliation, and our data suggest that the differentiation potential of basal cells at the outset is a more critical factor in air-liquid interface culture outcome. Detailed information on the constituents of the differentiation media was only available from one of the four manufacturers, a factor that may have profound implications in the interpretation of some research studies

Connecting Health and Technology (CHAT): protocol of a randomized controlled trial to improve nutrition behaviours using mobile devices and tailored text messaging in young adults

Background: Increasing intakes of fruits and vegetables intake, in tandem with reducing consumption of energy-dense and nutrient poor foods and beverages are dietary priorities to prevent chronic disease. Although most adults do not eat enough fruit and vegetables, teenagers and young adults tend to have the lowest intakes. Young adults typically consume a diet which is inconsistent with the dietary recommendations. Yet little is known about the best approaches to improve dietary intakes and behaviours among this group. This randomised controlled trial aims to evaluate the effectiveness of using a mobile device to assess dietary intake, provide tailored dietary feedback and text messages to motivate changes in fruit, vegetable and junk food consumption among young adults

Deteriorating health satisfaction among immigrants from Eastern Europe to Germany

BACKGROUND: Migrants from Eastern Europe constitute more than 5% of Germany's population. Since population health in their countries of origin is poor their health status upon arrival may be worse than that of the native-born German population (hypothesis H1). As a minority, they may be socio-economically disadvantaged (H2), and their health status may deteriorate quickly (H3). METHODS: We compared data from 1995 and 2000 for immigrants from Eastern Europe (n = 353) and a random sample of age-matched Germans (n = 2, 824) from the German Socioeconomic Panel. We tested H1-3 using health satisfaction, as a proxy for health status, and socioeconomic indicators. We compared changes over time within groups, and between immigrants and Germans. We assessed effects of socio-economic status and being a migrant on declining health satisfaction in a regression model. RESULTS: In 1995, immigrants under 55 years had a significantly higher health satisfaction than Germans. Above age 54, health satisfaction did not differ. By 2000, immigrants' health satisfaction had declined to German levels. Whereas in 1995 immigrants had a significantly lower SES, differences five years later had declined. In the regression model, immigrant status was much stronger associated with declining health satisfaction than low SES. CONCLUSION: In contrast to H1, younger immigrants had an initial health advantage. Immigrants were initially socio-economically disadvantaged (H2), but their SES improved over time. The decrease in health satisfaction was much steeper in immigrants and this was not associated with differences in SES (H3). Immigrants from Eastern Europe have a high risk of deteriorating health, in spite of socio-economic improvements

Deteriorating health satisfaction among immigrants from Eastern Europe to Germany

BACKGROUND: Migrants from Eastern Europe constitute more than 5% of Germany's population. Since population health in their countries of origin is poor their health status upon arrival may be worse than that of the native-born German population (hypothesis H1). As a minority, they may be socio-economically disadvantaged (H2), and their health status may deteriorate quickly (H3). METHODS: We compared data from 1995 and 2000 for immigrants from Eastern Europe (n = 353) and a random sample of age-matched Germans (n = 2, 824) from the German Socioeconomic Panel. We tested H1-3 using health satisfaction, as a proxy for health status, and socioeconomic indicators. We compared changes over time within groups, and between immigrants and Germans. We assessed effects of socio-economic status and being a migrant on declining health satisfaction in a regression model. RESULTS: In 1995, immigrants under 55 years had a significantly higher health satisfaction than Germans. Above age 54, health satisfaction did not differ. By 2000, immigrants' health satisfaction had declined to German levels. Whereas in 1995 immigrants had a significantly lower SES, differences five years later had declined. In the regression model, immigrant status was much stronger associated with declining health satisfaction than low SES. CONCLUSION: In contrast to H1, younger immigrants had an initial health advantage. Immigrants were initially socio-economically disadvantaged (H2), but their SES improved over time. The decrease in health satisfaction was much steeper in immigrants and this was not associated with differences in SES (H3). Immigrants from Eastern Europe have a high risk of deteriorating health, in spite of socio-economic improvements

Elevated myeloid-derived suppressor cells in pancreatic, esophageal and gastric cancer are an independent prognostic factor and are associated with significant elevation of the Th2 cytokine interleukin-13

We undertook a comprehensive analysis of circulating myeloid-derived suppressor cells (MDSCs) and T regulatory cells (Tregs) in pancreatic, esophageal and gastric cancer patients and investigated whether MDSCs are an independent prognostic factor for survival. We evaluated a series of plasma cytokines and in particular re-evaluated the Th2 cytokine interleukin-13 (IL-13). Peripheral blood was collected from 131 cancer patients (46 pancreatic, 60 esophageal and 25 gastric) and 54 healthy controls. PBMC were harvested with subsequent flow cytometric analysis of MDSC (HLADR− Lin1low/− CD33+ CD11b+) and Treg (CD4+ CD25+ CD127low/− FoxP3+) percentages. Plasma IL-2, IL-4, IL-5, IL-6, IL-10, IL-12 (p70), IL-13, IL-17, G-CSF, IFN-γ, TNF-α and VEGF levels were analyzed by the Bio-Plex cytokine assay. Plasma arginase I levels were analyzed by ELISA. MDSCs and Tregs were statistically significantly elevated in pancreatic, esophageal and gastric cancer compared with controls, and MDSC numbers correlated with Treg levels. Increasing MDSC percentage was associated with increased risk of death, and in a multivariate analysis, MDSC level was an independent prognostic factor for survival. A unit increase in MDSC percentage was associated with a 22% increased risk of death (hazard ratio 1.22, 95% confidence interval 1.06–1.41). Arginase I levels were also statistically significantly elevated in upper gastrointestinal cancer patients compared with controls. There was Th2 skewing for cytokine production in all three diseases, and importantly there were significant elevations of the pivotal Th2 cytokine interleukin-13, an increase that correlated with MDSC levels

Peripheral blood mononuclear cells from neovascular age-related macular degeneration patients produce higher levels of chemokines CCL2 (MCP-1) and CXCL8 (IL-8)

Flow cytometry analysis of PBMCs. PBMCs were first divided into CD11b+CD3−, CD11b−CD3+ and CD11b−CD3− cells (A) and the average percentage of all samples (n = 55) was analysed before and after stimulation with PMA/ionomycin (B). Figure S2. Percentage of total IL-4 and IL-10 producing PBMCs and percentage of CD11b−CD3+ IL-17A and IFNγ producing PBMCs (almost all of IL-17A and IFNγ producing PBMCs were CD11b−CD3+) from controls and nAMD patients under non-stimulated culture conditions and after stimulation with PMA/ionomycin. Controls n = 27, nAMD = 28; mean + SEM are shown. (PDF 413 kb

Design and evaluation protocol of "FATaintPHAT", a computer-tailored intervention to prevent excessive weight gain in adolescents

<p>Abstract</p> <p>Background</p> <p>Computer tailoring may be a promising technique for prevention of overweight in adolescents. However, very few well-developed, evidence-based computer-tailored interventions are available for this target group. We developed and evaluated a computer-tailored intervention for adolescents targeting energy balance-related behaviours: i.e. consumption of snacks, sugar-sweetened beverages, fruit, vegetables, and fibre, physical activity, and sedentary behaviours. This paper describes the planned development of a school-based computer-tailored intervention aimed at improving energy balance-related behaviours in order to prevent excessive weight gain in adolescents, and the protocol for evaluating this intervention.</p> <p>Methods/design</p> <p>Intervention development: Informed by the Precaution Adoption Process Model and the Theory of Planned Behaviour, the computer-tailored intervention provided feedback on personal behaviour and suggestions on how to modify it. The intervention (VETisnietVET translated as 'FATaintPHAT') has been developed for use in the first year of secondary school during eight lessons.</p> <p>Evaluation design: The intervention will be evaluated in a cluster-randomised trial including 20 schools with a 4-months and a 2-years follow-up. Outcome measures are BMI, waist circumference, energy balance-related behaviours, and potential determinants of these behaviours. Process measures are appreciation of and satisfaction with the program, exposure to the program's content, and implementation facilitators and barriers measured among students and teachers.</p> <p>Discussion</p> <p>This project resulted in a theory and evidence-based intervention that can be implemented in a school setting. A large-scale randomised controlled trial with a short and long-term follow-up will provide sound statements about the effectiveness of this computer-tailored intervention in adolescents.</p> <p>Trial Registration</p> <p>ISRCTN15743786</p

Airway response to respiratory syncytial virus has incidental antibacterial effects.

RSV infection is typically associated with secondary bacterial infection. We hypothesise that the local airway immune response to RSV has incidental antibacterial effects. Using coordinated proteomics and metagenomics analysis we simultaneously analysed the microbiota and proteomes of the upper airway and determined direct antibacterial activity in airway secretions of RSV-infected children. Here, we report that the airway abundance of Streptococcus was higher in samples collected at the time of RSV infection compared with samples collected one month later. RSV infection is associated with neutrophil influx into the airway and degranulation and is marked by overexpression of proteins with known antibacterial activity including BPI, EPX, MPO and AZU1. Airway secretions of children infected with RSV, have significantly greater antibacterial activity compared to RSV-negative controls. This RSV-associated, neutrophil-mediated antibacterial response in the airway appears to act as a regulatory mechanism that modulates bacterial growth in the airways of RSV-infected children