PYRIN-containing Apaf-1-like proteins (PYPAFs) are a recently identi¢ed family of proteins thought to function in apoptotic and in£ammatory signaling pathways. PYPAF1 and PYPAF7 proteins have been found to assemble with the PYRIN^CARD protein ASC and coordinate the activation of NF-U UB and pro-caspase-1. To determine if other PYPAF family members function in pro-in£ammatory signaling pathways, we screened ¢ve other PYPAF proteins (PYPAF2, PYPAF3, PY-PAF4, PYPAF5 and PYPAF6) for their ability to activate NF-U UB and pro-caspase-1. Co-expression of PYPAF5 with ASC results in a synergistic activation of NF-U UB and the recruitment of PYPAF5 to punctate structures in the cytoplasm. The expression of PYPAF5 is highly restricted to granulocytes and T-cells, indicating a role for this protein in in£ammatory signaling. In contrast, PYPAF2, PYPAF3, PYPAF4 and PYPAF6 failed to colocalize with ASC and activate NF-U UB. PYPAF5 also synergistically activated caspase-1-dependent cytokine processing when co-expressed with ASC. These ¢ndings suggest that PYPAF5 functions in immune cells to coordinate the transduction of pro-in£ammatory signals to the activation of NF-U UB and pro-caspase-1.

Rationale: Vitamin D insufficiency (a serum 25(OH)D ,30 ng/ml) has been associated with severe asthma exacerbations, but this could be explained by underlying racial ancestry or disease severity. Little is known about vitamin D and asthma in Puerto Ricans. Objectives: To examine whether vitamin D insufficiency is associated with severe asthma exacerbations in Puerto Rican children, independently of racial ancestry, atopy, and time outdoors. Methods: A cross-sectional study was conducted of 560 children ages 6-14 years with (n ¼ 287) and without (n ¼ 273) asthma in San Juan, Puerto Rico. We measured plasma vitamin D and estimated the percentage of African racial ancestry among participants using genomewide genotypic data. We tested whether vitamin D insufficiency is associated with severe asthma exacerbations, lung function, or atopy (greater than or equal to one positive IgE to allergens) using logistic or linear regression. Multivariate models were adjusted for African ancestry, time outdoors, atopy, and other covariates. Measurements and Main Results: Vitamin D insufficiency was common in children with (44%) and without (47%) asthma. In multivariate analyses, vitamin D insufficiency was associated with higher odds of greater than or equal to one severe asthma exacerbation in the prior year (odds ratio [OR], 2.6; 95% confidence interval [CI], 1.5-4.9; P ¼ 0.001) and atopy, and a lower FEV 1 /FVC in cases. After stratification by atopy, the magnitude of the association between vitamin D insufficiency and severe exacerbations was greater in nonatopic (OR, 6.2; 95% CI, 2-21.6; P ¼ 0.002) than in atopic (OR, 2; 95% CI, 1-4.1; P ¼ 0.04) cases. Conclusions: Vitamin D insufficiency is associated with severe asthma exacerbations in Puerto Rican children, independently of racial ancestry, atopy, or markers of disease severity or control.Keywords: vitamin D; asthma exacerbations; Puerto Ricans; childhood Vitamin D insufficiency (a serum level of 25(OH)D ,30 ng/ml) is common among children in the United States mainland. For example, a nationwide study of 2,759 U.S. children ages 6-11 years found vitamin D insufficiency in approximately 62% of nonHispanic whites, approximately 86% of Hispanics, and approximately 96% of non-Hispanic blacks (1). Vitamin D insufficiency has been associated with increased asthma morbidity, particularly severe disease exacerbations, in observational studies of children of school age in Costa Rica (2) and North America (3).Although current evidence from observational (2-5) and experimental studies (6-9) suggests that vitamin D insufficiency leads to severe asthma exacerbations or increased asthma morbidity in childhood (10), no study of vitamin D and asthma morbidity has accounted for objectively assessed racial ancestry and time spent outdoors. This is critical to exclude whether vitamin D insufficiency is associated with severe asthma exacerbations or asthma morbidity through "reverse causation" (e.g., more severe asthma leading to reduced time outdoors and thus decreased exposure to sunlight ...

Background Puerto Ricans and African Americans share a significant proportion of African ancestry. Recent findings suggest that African ancestry influences lung function in African American adults. Objective To examine whether a greater proportion of African ancestry is associated with lower FEV1 and FVC in Puerto Rican children, independently of socioeconomic status (SES), healthcare access or key environmental/lifestyle (EL) factors. Methods Cross-sectional case-control study of 943 Puerto Rican children ages 6 to 14 years with (n=520) and without (n=423) asthma (defined as physician-diagnosed asthma and wheeze in the prior year) living in Hartford (CT, n=383) and San Juan (PR, n=560). We estimated the percentage of African racial ancestry in study participants using genome-wide genotypic data. We tested whether African ancestry is associated with FEV1 and FVC using linear regression. Multivariate models were adjusted for indicators of SES and healthcare, and selected EL exposures. Results After adjustment for household income and other covariates, each 20% increment in African ancestry was significantly associated with lower pre-bronchodilator(BD) FEV1 (−105 ml, 95% confidence interval [CI] = −159 ml to −51 ml, P <0.001) and FVC (−133 ml, 95% CI −197 ml to −69 ml, P <0.001), and post-BD FEV1 (−152 ml, 95% CI=−210 ml to −94 ml, P <0.001) and FVC (−145 ml, 95% CI= −211 to −79 ml, P <0.001) in children with asthma. Similar but weaker associations were found for pre- and post-BD FEV1 (change for each 20% increment in African ancestry= −78 ml, 95% CI= −131 to −25 ml, P=0.004), and for post-BD FVC among children without asthma. Conclusions Genetic and/or EL factors correlated with African ancestry may influence childhood lung function in Puerto Ricans.

Background Gene by environment interaction (G × E) studies utilizing GWAS data are often underpowered after adjustment for multiple comparisons. Differential gene expression, in response to the exposure of interest, may capture the most biologically relevant genes at the genome-wide level. Methods We used differential genome-wide expression profiles from the Home Allergens and Asthma Birth cohort in response to Der f 1 allergen (sensitized vs. non-sensitized) to inform a G × E study of dust mite exposure and asthma severity. Polymorphisms in differentially expressed genes were identified in GWAS data from CAMP, a clinical trial in childhood asthmatics. Home dust mite allergen (< or ≥ 10µg/g dust) was assessed at baseline, and (≥ 1) severe asthma exacerbation (emergency room (ER) visit or hospitalization for asthma in the first trial year) served as the disease severity outcome. The Genetics of Asthma in Costa Rica (GACRS) study, and a Puerto Rico/Connecticut asthma cohortwere used for replication. Results IL-9, IL-5 and PRG2 expression was up-regulated in Der f 1 stimulated PBMCs from dust mite sensitized individuals (adj. p value <0.04). IL-9 polymorphisms (rs11741137, rs2069885, rs1859430) showed evidence for interaction with dust mite in CAMP (p=0.02 to 0.03), with replication in GACRS (p=0.04). Subjects with the dominant genotype for these IL-9 polymorphisms were more likely to report a severe asthma exacerbation if exposed to elevated dust mite. Conclusions Genome-wide differential gene expression in response to dust mite allergen identified IL-9, a biologically plausible gene target that may interact with environmental dust mite to increase severe asthma exacerbations in children.