PBMCs were placed in round-bottom 96-well plates (approximately 105 cells per well), stained for 30 min at 4 °C, washed twice with stain buffer with centrifugation at 250 g for 5 min, resuspended in 100 μL stain buffer and analyzed immediately. Monocytes were selectively gated based on their characteristic forward scatter and side Galunisertib price scatter properties.

The expression of CD11b, CD31, CD62L and CD49d on monocytes was quantified as percentage of positive cells from each sample. Associations between the indoor and outdoor pollutant levels were assessed by Pearson correlation coefficients. Linear regression models with the Generalized Estimating Equation approach (GEE) were used to estimate the association between log-transformed health outcomes and indoor and outdoor exposure variables, accounting for correlation between individuals living at the same address. Metformin Separate models were fitted for each outcome, adjusted for age, gender,

BMI and in sensitivity analyses for intake of vasoactive drugs or statins or use of candles as categorical variable. Additionally, the associations between the exposure and MVF were assessed for a subgroup of study participants who did not take any drugs (n = 65), adjusted for age, gender, and BMI. Furthermore, we included adjustment for the time the home was unoccupied (on average 20% of the total time) as an estimate of time spent outside in sensitivity analyses of the significant associations found. Results were expressed as percentage change with 95% confidence intervals of an outcome per increase in a pollutant’s interquartile range (IQR) concentration. We used the IQRs in the analysis of the indoor and the outdoor data pollutant to allow direct comparison of effect estimates. A value of p ≤ 0.05 was considered statistically significant. Leukotriene-A4 hydrolase Analyses were performed using STATA software (version 12, StataCorp LP, College Station, Texas, USA). Table 2 outlines the results from the 2-day indoor air monitoring of the 58 residences for PNC, mean particle diameter PM2.5 and the level of

endotoxin, fungi and bacteria levels in dust collected for 4 weeks. The levels of the indoor PNC have recently been reported (Bekö et al., 2013). The ambient air PNC, mean particle diameter, PM2.5 and PM10 concentrations, monitored at an urban background station in the same 2-day period preceding the measurements of health outcomes are also summarized in Table 2. There was a significant positive correlation between indoor PNC and PM2.5, whereas there were inverse positive correlations between indoor PNC and outdoor PM2.5 and PM10 levels, although these were not significant. The average indoor PNC levels over the whole monitoring period were strongly associated with the estimated exposure related to candle burning as source events.