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Moisture-damaged buildings are associated with respiratory symptoms and underlying diseases among building occupants, but the causative agent(s) remain a mystery. We first identified specific fungal and bacterial taxa in classrooms with moisture damage in Finnish and Dutch primary schools. We then investigated associations of the identified moisture damage indicators with respiratory symptoms in more than 2700 students. Finally, we explored whether exposure to specific taxa within the indoor microbiota may explain the association between moisture damage and respiratory health. Schools were assessed for moisture damage through detailed inspections, and the microbial composition of settled dust in electrostatic dustfall collectors was determined using marker-gene analysis. In Finland, there were several positive associations between particular microbial indicators (diversity, richness, individual taxa) and a respiratory symptom score, while in the Netherlands, the associations tended to be mostly inverse and statistically non-significant. In Finland, abundance of the Sphingomonas bacterial genus and endotoxin levels partially explained the associations between moisture damage and symptom score. A few microbial taxa explained part of the associations with health, but overall, the observed associations between damage-associated individual taxa and respiratory health were limited.

In vitro models mimicking the human respiratory system are essential when investigating the toxicological effects of inhaled indoor air particulate matter (PM). We present a pulmonary cell culture model for studying indoor air PM toxicity. We exposed normal human bronchial epithelial cells, grown on semi‐permeable cell culture membranes, to four doses of indoor air PM in the air‐liquid interface. We analyzed the chemokine interleukin‐8 concentration from the cell culture medium, protein concentration from the apical wash, measured tissue electrical resistance, and imaged airway constructs using light and transmission electron microscopy. We sequenced RNA using a targeted RNA toxicology panel for 386 genes associated with toxicological responses. PM was collected from a non‐complaint residential environment over 1 week. Sample collection was concomitant with monitoring size‐segregated PM counts and determination of microbial levels and diversity. PM exposure was not acutely toxic for the cells, and we observed up‐regulation of 34 genes and down‐regulation of 17 genes when compared to blank sampler control exposure. The five most up‐regulated genes were related to immunotoxicity. Despite indications of incomplete cell differentiation, this model enabled the comparison of a toxicological transcriptome associated with indoor air PM exposure.

Additional file 8 Figure S4. Viability (A), Comet assay (B) and IL-8 secretion (C) of A549 control cells after exposed one hour at incubator with CO2, incubator without CO2 or Tox-ALI to clean air. Each bar shows mean ± SEM, n = 3.

Additional file 4 Figure S2. Size of the particles measured by SMPS per day to day. The geometric mean diameters are shown with the solid lines, while GSDs are represented with dashed lines. (average of 3 experiments)

Additional file 7 Table S3. Inorganic elements from spruce and pine combustion emissions. Organic compounds were sampled over 4-h exposure. Concentrations are calculated as averages from three test cycles ± standard deviation.

Additional file 9 Table S4. Top 5 significant ingenuity canonical pathways of spruce aerosol exposure.

Additional file 5 Figure S3. Size of the particles measured by SMPS. The geometric mean diameters (a) and mass median diameters (b) are shown with the solid lines, while GSDs are represented with dashed lines. Average of 3 experiments.

Additional file 11 Figure S5. Area under curve (AUC) calculated from exposed A549 cells from three different timepoints of H2DCFDA measurements. Each bar shows mean ± SEM, n = 3.

Exposure to moisture-damaged indoor environments is associated with adverse respiratory health effects, but responsible factors remain unidentified. In order to explore possible mechanisms behind these effects, the oxidative capacity and hemolytic activity of settled dust samples (n = 25) collected from moisture-damaged and non-damaged schools in Spain, the Netherlands, and Finland were evaluated and matched against the microbial content of the sample. Oxidative capacity was determined with plasmid scission assay and hemolytic activity by assessing the damage to isolated human red blood cells. The microbial content of the samples was measured with quantitative PCR assays for selected microbial groups and by analyzing the cell wall markers ergosterol, muramic acid, endotoxins, and glucans. The moisture observations in the schools were associated with some of the microbial components in the dust, and microbial determinants grouped together increased the oxidative capacity. Oxidative capacity was also affected by particle concentration and country of origin. Two out of 14 studied dust samples from moisture-damaged schools demonstrated some hemolytic activity. The results indicate that the microbial component connected with moisture damage is associated with increased oxidative stress and that hemolysis should be studied further as one possible mechanism contributing to the adverse health effects of moisture-damaged buildings.

Aiming to identify factors causing the adverse health effects associated with moisture damaged indoor environments, we analyzed immunotoxicological potential of settled dust from moisture damaged and reference schools in relation to their microbiological composition. Mouse RAW264.7 macrophages were exposed to settled dust samples (n= 25) collected from moisture damaged and reference schools in Spain, The Netherlands and Finland. After exposure, we analyzed production of inflammatory markers [nitric oxide (NO), tumor necrosis factor (TNF)α, interleukin (IL)-6 and macrophage inflammatory protein (MIP)2] as well as mitochondrial activity, viability, apoptosis and cell cycle arrest. Furthermore, particle counts, concentration of selected microbial groups as well as chemical markers ergosterol, 3-hydroxy fatty acids, muramic acid, endotoxins and glucans were measured as markers of exposure. Dust from moisture damaged schools in Spain and The Netherlands induced stronger immunotoxicological responses compared to samples from reference schools; the responses to Finnish samples were generally lower with no difference between the schools. In multivariate analysis, IL-6 and apoptosis responses were most strongly associated with moisture status of the school. The measured responses correlated with several microbial markers and numbers of particles, but the most important predictor of the immunotoxicological potential of settled dust was muramic acid concentration, a marker of Gram positive bacteria. This article is protected by copyright. All rights reserved.

Aiming to identify factors causing the adverse health effects associated with moisture damaged indoor environments, we analyzed immunotoxicological potential of settled dust from moisture damaged and reference schools in relation to their microbiological composition. Mouse RAW264.7 macrophages were exposed to settled dust samples (n= 25) collected from moisture damaged and reference schools in Spain, The Netherlands and Finland. After exposure, we analyzed production of inflammatory markers [nitric oxide (NO), tumor necrosis factor (TNF)α, interleukin (IL)-6 and macrophage inflammatory protein (MIP)2] as well as mitochondrial activity, viability, apoptosis and cell cycle arrest. Furthermore, particle counts, concentration of selected microbial groups as well as chemical markers ergosterol, 3-hydroxy fatty acids, muramic acid, endotoxins and glucans were measured as markers of exposure. Dust from moisture damaged schools in Spain and The Netherlands induced stronger immunotoxicological responses compared to samples from reference schools; the responses to Finnish samples were generally lower with no difference between the schools. In multivariate analysis, IL-6 and apoptosis responses were most strongly associated with moisture status of the school. The measured responses correlated with several microbial markers and numbers of particles, but the most important predictor of the immunotoxicological potential of settled dust was muramic acid concentration, a marker of Gram positive bacteria. This article is protected by copyright. All rights reserved.

Cytokine concentrations in biological fluids are widely used markers for activation of immunological processes. Confirming the reproducibility of measurements is important, especially in longitudinal or multicenter studies where time between analyses or different analyzing laboratories increases the intra-assay variability. In this study, the reproducibility of the cytokine analysis conducted with different assay platforms was studied by comparing the results of two cytokines [interleukin (IL)-6 in serum and nasal lavage fluid (NAL) and IL-8 in NAL] analyzed with Meso Scale Discovery (MSD) ultra-sensitive single and multiplex assay kits (n=76). In addition, the difference in cytokine levels between two biological sample matrices was studied by comparing the results of altogether 9 cytokines [IL-6, IL-2, IL-8, IL12p70, IL-1β, granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon (IFN)γ, IL-10 and tumor necrosis factor (TNF)α] measured from serum and NAL of the same study subjects (n=460).The results show that the cytokine concentrations analyzed with single and multiplex assays are concordant but not equal. Comparison of the different matrices revealed that cytokine concentrations in serum do not correspond with concentrations detected in nasal lavage fluid.It can be concluded that comparability of the results from single and multiplex analysis of cytokines is high, but the concentrations should not be compared directly with each other. The differences between concentrations analyzed from serum and nasal lavage fluid indicate that the levels are specific for each matrix and represent distinct immunological conditions.

Green carella viitataan eläinten, luonnon ja maaseudun ympäristöjen hyvinvointia tuoviin vaikutuksiin sekä niiden hyödyntämiseen hyvinvointipalveluiden tuotannossa. Green careen liittyvä yrittäjyys nähdään sosiaalisena innovaationa, jonka syntymiseen ovat olleet vaikuttamassa sekä maatalouden että sosiaali- ja terveydenhuollon kohtaamat rakennemuutokset. Green caren käsite on Suomessa nostettu esille viime vuosina etenkin hyvinvointialan pienyrittäjyyttä kehittävien hankkeiden yhteydessä. Tutkielma tarkastelee sitä, millaisia kokemuksia sosiaali- ja terveysalan ammattilaisilla on green care -yrittäjyyden käynnistymisestä Suomessa. Terveyden ja hyvinvoinnin asiantuntijoina sosiaali- ja terveysalan ammattilaiset muodostavat keskeisen ja tärkeän ryhmän green care -yrittäjissä. Tutkielman tavoitteena on lisätä tietoa ammattilaisten erityispiirteistä yhtenä green care -yrittäjien ryhmänä. Empiirisen aineiston keruu on tehty haastattelemalla kuutta green caren kehittämishankkeisiin osallistunutta sosiaali- ja terveysalan ammattilaista, jotka toimivat green care -yrittäjinä tai suunnittelivat alan yrityksen perustamista. Aineiston tarkastelussa hyödynnetään työurien tutkimuksessa käytössä olevia näkökulmia ja käsitteitä. Kokemuksia green care -yrittäjyydestä valaistaan tarkastelemalla yrittäjien työurien kulkua. Lähestymistapa on narratiivinen, jolloin huomio kiinnittyy siihen, miten haastateltavat rakentavat työuraansa kertomuksena. Haastatteluaineiston analyysi toteutettiin sisällönanalyysillä teemoittelun periaatteita noudattaen. Narratiivinen viitekehys korostaa haastateltavien omia kokemuksia, mutta valottaa myös kokemisen ja kertomisen yhteyttä laajempiin kulttuurisiin ja sosiaalisiin konteksteihin. Green care -yrittäjien uranarratiivit nojasivat sosiaali- ja terveysalan ihanteisiin ja tarinankertomisen malleihin, mutta tämän lisäksi niissä oli havaittavissa myös ajatuksia työurien yksilöllistymisestä. Sosiaali- ja terveysalan ammattilaisten kokemuksissa green care -yrittäjyys nähtiin eettisempänä hoivan tuottamisena, ammatillisena erikoistumisena sekä sellaisen uudenlaisen uravaihtoehdon etsimisenä, jossa keskeiseksi muodostui omiin vahvuuksiin ja mielenkiinnon kohteisiin nojaavan työn ja yrittäjyyden luominen.

Additional file 6 Table S2. All measured organic compounds from wood combustion emissions. Organic compounds were sampled over 4-h exposure. Concentrations are calculated as averages from three test cycles ± standard deviation.

Additional file 1: Table S1. Calibration and quantification mixtures of isotope labelled internal standards and calibration standards.

The human respiratory muco-ciliary epithelium consists of different cell types, connected by tight junctions; forming a protective barrier against the external environment. The epithelium is attached to a basement membrane through which necessary nutrients are delivered. Apart from a thin layer of mucus, the apical part of the bronchial epithelium is exposed to the ambient air. Mimicking these conditions through the use of 3-dimensional (3-D) filter-well membrane technology, a functional and fully-differentiated muco-ciliary phenotype in vitro cell culture model can be obtained. By providing the in vivo-like pulmonary microenvironment primary bronchial cells are stimulated to differentiate into a pseudo-stratified epithelium containing, basal, ciliated, Clara, goblet, intermediate and serous cells. In addition to this in vivo-like morphology, the cultures attain physiological properties akin to the human bronchial epithelium. Cells form tight junctions, enabling the whole culture to function as a barrier, providing trans-epithelial electrical resistance (TEER). Day 15 of ALI culture conditions results in beating cilia that are fully-grown and spread the mucus produced by the goblet cells. Unlike ‘conventional’ cell cultures, where tested substances need to be suspended in submerged culture media, our 3-D model of the human bronchial epithelium enables in vitro toxicology testing of apically deposited aerosols and/or solubilised substances onto mucus protected cell surfaces. This mimics in vivo deposition through inhalation. Moreover, we observe the tissue (i.e. multi-cellular) response, rather than the effect of submerged cell monolayers (i.e. single cell type). The additional protective mechanisms, such as apical mucus, traps inhaled substances and the tight junctions (i.e. barrier function) provide a more in vivo-like response when compared to submerged monolayer cultures. Our 3-D model of the human bronchial epithelium maintains the in vivo complexity observed in the in-situ human lung and is a reliable in vitro tool that can be employed to obtain robust predictive in vivo responses for human endpoint data.

Additional file 6 Table S2. All measured organic compounds from wood combustion emissions. Organic compounds were sampled over 4-h exposure. Concentrations are calculated as averages from three test cycles ± standard deviation.

Exposure to indoor air in moisture damaged buildings is associated with deteriorating respiratory health, assumedly due to emissions from microbial growth and wet building materials. Previous studies of toxicological effects of mouldy house microbes have indicated that inflammation and cell death are important mechanisms. Aiming to gain further insight into function of respiratory epithelia, we studied the responses of normal human bronchial epithelium (NHBE) model after exposure to settled dust samples (n=9) collected from moisture damaged and reference schools in Spain, The Netherlands and Finland. The results were compared with immunotoxicological potential of the same samples in mouse RAW264.7 macrophage model. The effects of exposure on the NHBE model was assessed by measuring trans epithelial electric resistance (TEER) of the culture, changes in mucus production (Bradford assay), cell toxicity (ATP assay), cytokine levels in the culture media (ELISA) as well as morphological changes in cultured cells (light and scanning electron microscopy). The results showed that exposure to dust from moisture damaged schools was capable of inducing both increased TEER and mucus production and in some instances diminished TEER values indicating deterioration of tissue integrity and cell death. Similarly with the results from mouse macrophage model, the samples from The Netherlands and Spain were more toxic compared to samples from Finland. The findings suggest that the defence mechanisms present in respiratory epithelia are activated by dust from moisture damaged buildings, and exposure to high doses may damage the affected lung tissue.

Additional file 3 Figure S1. Physical properties in the diluted exposure exhaust per day to day, for spruce (A) and pine (B) experiments. Parts A and C show average of 3 experiment days total suspended particulate mass (TSP), lung deposited surface area (LDSA) and particle number concentration (PNC) during exposures.

Additional file 1: Table S1. Calibration and quantification mixtures of isotope labelled internal standards and calibration standards.

Additional file 3 Figure S1. Physical properties in the diluted exposure exhaust per day to day, for spruce (A) and pine (B) experiments. Parts A and C show average of 3 experiment days total suspended particulate mass (TSP), lung deposited surface area (LDSA) and particle number concentration (PNC) during exposures.

Exposure to moisture damaged indoor environment is associated with adverse respiratory health effects, but responsible factors remain unidentified. In order to elucidate the mechanism behind these effects, Reactive Oxidative Stress (ROS)-generating capacity of settled dust samples (n=25) collected from moisture damaged and reference schools in Spain, The Netherlands and Finland was evaluated. In addition, the results were compared with immunotoxicological endpoints analysed with an in vitro model. ROS capacity was assessed with a plasmid scission assay (PSA), which determines the dose able to damage 50 % of DNA from a plasmid sensitive to ROS (TD50 value). In addition, immunotoxicological endpoints such as production of inflammatory markers as well as mitochondrial activity, viability, apoptosis and cell cycle arrest were analysed in vitro using mouse RAW264.7 macrophages as a model. \ud The average TD50 values showed that samples from moisture damaged schools in Spain and The Netherlands had higher ROS capacity compared to samples from reference schools, whereas the capacity of Finnish samples was lower with no clear difference between the schools. The results were in line with the findings of an in vitro model showing significant geographical differences and a trend for higher potency of samples from moisture damaged environments in two out of three countries. The results indicate that ROS producing capacity of indoor dust tends to be higher in moisture damaged buildings, but geographical differences and high variance confounds the differentiation between moisture damaged and reference environments.