Mapping a Conformational Epitope of Hemagglutinin A Using Native Mass Spectrometry and Ultraviolet Photodissociation.

As the importance of effective vaccines and the role of protein therapeutics in the drug industry continue to expand, alternative strategies to characterize protein complexes are needed. Mass spectrometry (MS) in conjunction with enzymatic digestion or chemical probes has been widely used for mapping binding epitopes at the molecular level. However, advances in instrumentation and application of activation methods capable of accessing higher energy dissociation pathways have recently allowed direct analysis of protein complexes. Here we demonstrate a workflow utilizing native MS and ultraviolet photodissociation (UVPD) to map the antigenic determinants of a model antibody-antigen complex involving hemagglutinin (HA), the primary immunogenic antigen of the influenza virus, and the D1 H1-17/H3-14 antibody which has been shown to confer potent protection to lethal infection in mice despite lacking neutralization activity. Comparison of sequence coverages upon UV photoactivation of HA and of the HA·antibody complex indicates the elimination of some sequence ions that originate from backbone cleavages exclusively along the putative epitope regions of HA in the presence of the antibody. Mapping the number of sequence ions covering the HA antigen versus the HA·antibody complex highlights regions with suppressed backbone cleavage and allows elucidation of unknown epitopes. Moreover, examining the observed fragment ion types generated by UVPD demonstrates a loss in diversity exclusively along the antigenic determinants upon MS/MS of the antibody-antigen complex. UVPD-MS shows promise as a method to rapidly map epitope regions along antibody-antigen complexes as novel antibodies are discovered or developed.

Accuracy Assessment of NLCD 2011 percent impervious cover for selected USA Metropolitan Areas.

The emergence of high-resolution land cover data has created the opportunity to assess the accuracy of impervious cover (IC) provided by the National Land Cover Database (NLCD). We assessed the accuracy of the 900 m2 NLCD2011 %IC for 18 metropolitan areas throughout the conterminous United States using reference data from 1 m2 land cover data developed as part of the United States Environmental Protection Agency's EnviroAtlas project. Agreement was assessed from two perspectives: 1) sensitivity to the size of the assessment unit used for the comparison, and 2) utility of NLCD %IC to serve as a proxy for high-resolution IC. The former perspective was considered because statistical relationships can be sensitive to assessment unit size and shape, and the latter perspective was considered because high resolution (reference) %IC data are not available nationwide. The utility of NLCD %IC as a proxy for the high resolution data was assessed for seven lattice (square) cell sizes ranging from 1 ha to 200 ha using four EnviroAtlas IC indicators: 1) %IC per 100 ha (1 km2); 2) %IC by Census block group; 3) %IC within a 15 m (radius) of the riparian zone, and; 4) %IC within a 50 m (radius) of the riparian zone. Agreement was quantified as per assessment unit deviation (NLCD %IC - reference %IC) and summarized as Mean Absolute Deviation (MAD) and Mean Deviation (MD) both within and across the 18 metropolitan areas. Ordinary least squares (OLS) regression (y = reference %IC and x = NLCD %IC) was also used to evaluate the quality of the NLCD %IC data. MAD was ≤ 5% for six of the seven lattice cell sizes. MAD was also ≤ 5% for Census block groups > 100 ha and for both riparian units. These results suggest that uncertainty attributable to the measurement of %IC was no greater than the uncertainty related to the effect of IC on aquatic resources that have been derived from studies of aquatic condition (e.g., benthic fauna) over a range of %IC. Overall, agreement was variable from one metropolitan area to the next. Agreement improved as assessment unit size increased and declined as the level of urbanization (NLCD %IC) increased. NLCD %IC tended to underestimate reference %IC overall, but NLCD %IC was sometimes greater than reference %IC in urbanized settings.

Linking land cover and water quality in New York City's water supply watersheds.

The Catskill/Delaware reservoirs supply 90% of New York City's drinking water. The City has implemented a series of watershed protection measures, including land acquisition, aimed at preserving water quality in the Catskill/Delaware watersheds. The objective of this study was to examine how relationships between landscape and surface water measurements change between years. Thirty-two drainage areas delineated from surface water sample points (total nitrogen, total phosphorus, and fecal coliform bacteria concentrations) were used in step-wise regression analyses to test landscape and surface-water quality relationships. Two measurements of land use, percent agriculture and percent urban development, were positively related to water quality and consistently present in all regression models. Together these two land uses explained 25 to 75% of the regression model variation. However, the contribution of agriculture to water quality condition showed a decreasing trend with time as overall agricultural land cover decreased. Results from this study demonstrate that relationships between land cover and surface water concentrations of total nitrogen, total phosphorus, and fecal coliform bacteria counts over a large area can be evaluated using a relatively simple geographic information system method. Land managers may find this method useful for targeting resources in relation to a particular water quality concern, focusing best management efforts, and maximizing benefits to water quality with minimal costs.