Scara1 deficiency impairs clearance of soluble Amyloid-β by mononuclear phagocytes and accelerates Alzheimer’s-like disease progression

In Alzheimer’s disease soluble amyloid beta (sAβ) causes synaptic dysfunction and neuronal loss. Receptors involved in clearance of sAβ are not known. Here we use shRNA screening and identify the scavenger receptor Scara1 as a receptor for sAβ expressed on myeloid cells. To determine the role of Scara1 in clearance of sAβ in vivo, we cross Scara1 null mice with PS1-APP mice, a mouse model of Alzheimer’s disease and generate PS1-APP- Scara1-deficient mice. Scara1 deficiency markedly accelerates Aβ accumulation leading to increased mortality. In contrast, pharmacological upregulation of Scara1 expression on mononuclear phagocytes increases Aβ clearance. This approach is a potential treatment strategy for Alzheimer’s disease

Scara1 deficiency impairs clearance of soluble Amyloid-β by mononuclear phagocytes and accelerates Alzheimer’s-like disease progression

In Alzheimer’s disease soluble amyloid beta (sAβ) causes synaptic dysfunction and neuronal loss. Receptors involved in clearance of sAβ are not known. Here we use shRNA screening and identify the scavenger receptor Scara1 as a receptor for sAβ expressed on myeloid cells. To determine the role of Scara1 in clearance of sAβ in vivo, we cross Scara1 null mice with PS1-APP mice, a mouse model of Alzheimer’s disease and generate PS1-APP- Scara1-deficient mice. Scara1 deficiency markedly accelerates Aβ accumulation leading to increased mortality. In contrast, pharmacological upregulation of Scara1 expression on mononuclear phagocytes increases Aβ clearance. This approach is a potential treatment strategy for Alzheimer’s disease

High-resolution profiling of homing endonuclease binding and catalytic specificity using yeast surface display

Experimental analysis and manipulation of protein–DNA interactions pose unique biophysical challenges arising from the structural and chemical homogeneity of DNA polymers. We report the use of yeast surface display for analytical and selection-based applications for the interaction between a LAGLIDADG homing endonuclease and its DNA target. Quantitative flow cytometry using oligonucleotide substrates facilitated a complete profiling of specificity, both for DNA-binding and catalysis, with single base pair resolution. These analyses revealed a comprehensive segregation of binding specificity and affinity to one half of the pseudo-dimeric interaction, while the entire interface contributed specificity at the level of catalysis. A single round of targeted mutagenesis with tandem affinity and catalytic selection steps provided mechanistic insights to the origins of binding and catalytic specificity. These methods represent a dynamic new approach for interrogating specificity in protein–DNA interactions

Two centuries of nitrogen dynamics: Legacy sources and sinks in the Mississippi and Susquehanna River Basins

© American Geophysical UnionGlobal flows of reactive nitrogen (N) have increased significantly over the last century in response to agricultural intensification and elevated levels of atmospheric deposition. Despite widespread implementation conservation measures, N concentrations in surface waters are often remaining steady or continuing to increase. Although such lack of response has been attributed to time lags associated with legacy N stores in subsurface reservoirs, it is unclear what the magnitudes of such stores are and how they are partitioned between shallow soil and deeper groundwater reservoirs. Here we have synthesized data to develop a 214year (1800-2014) trajectory of N inputs to the land surface of the continental U.S. We have concurrently developed a parsimonious, process-based model, Exploration of Long-tErM Nutrient Trajectories (ELEMeNT) that pairs this input trajectory with a travel time-based approach to simulate transport and retention along subsurface pathways. Using the model, we have reconstructed historic nitrate yields at the outlets of two major U.S. watersheds, the Mississippi River Basin (MRB) and Susquehanna River Basin (SRB). Our results show significant N loading above baseline levels in both watersheds before the widespread use of commercial N fertilizers, largely due to the conversion of forest and grassland to row crop agriculture. Model results also allow us to quantify the magnitudes of legacy N in soil and groundwater pools and to highlight the dominance of soil legacies in MRB and groundwater legacies in SRB. Approximately 55% and 18% of the current annual N loads in the MRB and SRB were found to be older than 10years of age.NSERC Discovery Grant; Ontario Early Researcher Awar