Conditionally reprogrammed asthmatic bronchial epithelial cells express lower FOXJ1 at terminal differentiation and lower IFNs following RV-A1 infection.

Primary bronchial epithelial cells (pBECs) obtained from donors have limited proliferation capacity. Recently, conditional reprogramming (CR) technique has overcome this and has provided the potential for extended passaging and subsequent differentiation of cells at air-liquid interface (ALI). However, there has been no donor-specific comparison of cell morphology, baseline gene expression, barrier function, and antiviral responses compared with their "parent" pBECs, especially cells obtained from donors with asthma. We, therefore, collected and differentiated pBECs at ALI from mild donors with asthma (n = 6) for the parent group. The same cells were conditionally reprogrammed and later differentiated at ALI. Barrier function was measured during the differentiation phase. Morphology and baseline gene expression were compared at terminal differentiation. Viral replication kinetics and antiviral responses were assessed following rhinovirus (RV) infection over 96 h. Barrier function during the differentiation phase and cell structural morphology at terminal differentiation appear similar in both parent and CR groups, however, there were elongated cell structures superficial to basal cells and significantly lower FOXJ1 expression in CR group. IFN gene expression was also significantly lower in CR group compared with parent asthma group following RV infection. The CR technique is a beneficial tool to proliferate pBECs over extended passages. Considering lower FOXJ1 expression, viral replication kinetics and antiviral responses, a cautious approach should be taken while choosing CR cells for experiments. In addition, as lab-to-lab cell culture techniques vary, the most appropriate technique must be utilized to best match individual cell functions and morphologies to address specific research questions and experimental reproducibility across the labs.

Line Transect Surveys Underdetect Terrestrial Mammals: Implications for the Sustainability of Subsistence Hunting.

Conservation of Neotropical game species must take into account the livelihood and food security needs of local human populations. Hunting management decisions should therefore rely on abundance and distribution data that are as representative as possible of true population sizes and dynamics. We simultaneously applied a commonly used encounter-based method and an infrequently used sign-based method to estimate hunted vertebrate abundance in a 48,000-km2 indigenous landscape in southern Guyana. Diurnal direct encounter data collected during three years along 216, four-kilometer -long transects consistently under-detected many diurnal and nocturnal mammal species readily detected through sign. Of 32 species analyzed, 31 were detected by both methods; however, encounters did not detect one and under-detected another 12 of the most heavily hunted species relative to sign, while sign under-detected 12 never or rarely collected species relative to encounters. The six most important game animals in the region, all ungulates, were not encountered at 11-40% of village and control sites or on 29-72% of transects where they were detected by sign. Using the sign methodology, we find that tapirs, one of the terrestrial vertebrates considered most sensitive to overexploitation, are present at many sites where they were never visually detected during distance sampling. We find that this is true for many other species as well. These high rates of under-detection suggest that behavioral changes in hunted populations may affect apparent occurrence and abundance of these populations. Accumulation curves (detection of species on transects) were much steeper for sign for 12 of 16 hunted species than for encounters, but that pattern was reversed for 12 of 16 species unhunted in our area. We conclude that collection of sign data is an efficient and effective method of monitoring hunted vertebrate populations that complements encounter and camera-trapping methods in areas impacted by hunting. Sign surveys may be the most viable method for large-scale, management-oriented studies in remote areas, particularly those focused on community-based wildlife management.

The spatial pattern of nitrogen cycling in the Adirondack Park, New York.

Maps of canopy nitrogen obtained through analysis of high-resolution, hyperspectral, remotely sensed images now offer a powerful means to make landscape-scale to regional-scale estimates of forest N cycling and net primary production (NPP). Moreover, recent research has suggested that the spatial variability within maps of canopy N may be driven by environmental gradients in such features as historic forest disturbance, temperature, species composition, moisture, geology, and atmospheric N deposition. Using the wide variation in these six features found within the diverse forest ecosystems of the 2.5 million ha Adirondack Park, New York, USA, we examined linkages among environmental gradients and three measures of N cycling collected during the 2003 growing season: (1) field survey of canopy N, (2) field survey of soil C:N, and (3) canopy N measured through analysis of two 185 x 7.5 km Hyperion hyperspectral images. These three measures of N cycling strongly related to forest type but related poorly to all other environmental gradients. Further analysis revealed that the spatial pattern in N cycling appears to have distinct inter- and intraspecific components of variability. The interspecific component, or the proportional contribution of species functional traits to canopy biomass, explained 93% of spatial variability within the field canopy N survey and 37% of variability within the soil C:N survey. Residual analysis revealed that N deposition accounted for an additional 2% of variability in soil C:N, and N deposition and historical forest disturbance accounted for an additional 2.8% of variability in canopy N. Given our finding that 95.8% of the variability in the field canopy N survey could be attributed to variation in the physical environment, our research suggests that remotely sensed maps of canopy N may be useful not only to assess the spatial variability in N cycling and NPP, but also to unravel the relative importance of their multiple controlling factors.

Foliar nitrogen responses to elevated atmospheric nitrogen deposition in nine temperate forest canopy species.

Despite its ecological importance, broad-scale use of foliar nitrogen as an indicator of ecosystem response to atmospheric N deposition has heretofore been obscured by its poorly understood intrinsic variability through time, space, and across species. We used a regional survey of foliar N conducted within a single growing season to observe that eight of nine major canopy tree species had increased foliar N in response to a gradient of N deposition in the Adirondack Park, New York. These results (1) add important foliar N evidence to support N saturation theory, (2) strongly reinforce the conclusion that N deposition is affecting the N status of forest ecosystems in the northeastern U.S., and (3) extend N saturation theory by identifying that temperate forest canopy species differ in their foliar N response to N deposition. Interestingly, species-specific differences were strongly related to two functional traits that arise from within-leaf allocations of N resources--leaf mass per area (LMA) and shade tolerance. Thus, combining species-specific knowledge of these functional traits with existing foliar N-centered remote sensing and ecosystem modeling approaches may provide a much-needed avenue to make broad-scale assessments of how persistently elevated rates of N deposition will continue to affect temperate forest ecosystems.