Defining the RBPome of primary T helper cells to elucidate higher-order Roquin-mediated mRNA regulation.

Post-transcriptional gene regulation in T cells is dynamic and complex as targeted transcripts respond to various factors. This is evident for the Icos mRNA encoding an essential costimulatory receptor that is regulated by several RNA-binding proteins (RBP), including Roquin-1 and Roquin-2. Here, we identify a core RBPome of 798 mouse and 801 human T cell proteins by utilizing global RNA interactome capture (RNA-IC) and orthogonal organic phase separation (OOPS). The RBPome includes Stat1, Stat4 and Vav1 proteins suggesting unexpected functions for these transcription factors and signal transducers. Based on proximity to Roquin-1, we select ~50 RBPs for testing coregulation of Roquin-1/2 targets by induced expression in wild-type or Roquin-1/2-deficient T cells. Besides Roquin-independent contributions from Rbms1 and Cpeb4 we also show Roquin-1/2-dependent and target-specific coregulation of Icos by Celf1 and Igf2bp3. Connecting the cellular RBPome in a post-transcriptional context, we find contributions from multiple RBPs to the prototypic regulation of mRNA targets by individual trans-acting factors.

Creating a Sustainable Palliative Care Education Conference for Healthcare Professionals.

An interprofessional conference that addressed basic palliative care knowledge and skills was implemented annually at a large, academic healthcare institution. The conference significantly influenced participant's self-perceived competencies in the delivery of quality palliative care. This conference led to a sustainable program of palliative care initiatives consisting of a yearly workshop for nurses focused on palliative care core competencies and a biennial interprofessional conference designed to facilitate networking and address significant topics in palliative care.

Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA.

The ubiquitously expressed RNA-binding proteins Roquin-1 and Roquin-2 are essential for appropriate immune cell function and postnatal survival of mice. Roquin proteins repress target mRNAs by recognizing secondary structures in their 3'-UTRs and by inducing mRNA decay. However, it is unknown if other cellular proteins contribute to target control. To identify cofactors of Roquin, we used RNA interference to screen ~1500 genes involved in RNA-binding or mRNA degradation, and identified NUFIP2 as a cofactor of Roquin-induced mRNA decay. NUFIP2 binds directly and with high affinity to Roquin, which stabilizes NUFIP2 in cells. Post-transcriptional repression of human ICOS by endogenous Roquin proteins requires two neighboring non-canonical stem-loops in the ICOS 3'-UTR. This unconventional cis-element as well as another tandem loop known to confer Roquin-mediated regulation of the Ox40 3'-UTR, are bound cooperatively by Roquin and NUFIP2. NUFIP2 therefore emerges as a cofactor that contributes to mRNA target recognition by Roquin.

Assessment of the historical trace metal contamination of sediments in the Elizabeth River, Virginia.

Two sediment cores (Southern Branch, PC-1, and Western Branch, WB-2) were taken from the highly industrialized Elizabeth River, Virginia. The concentrations of trace metals cadmium, cobalt, chromium, copper, nickel, lead and zinc, major elements iron, manganese and aluminum, organic carbon content and the specific surface area of the sediments were determined in each of the cores. Down-core variations in metals varied significantly in each core with maximum contamination events occurring at different times in different portions of the river. In PC-1, maximum metal concentrations were seen after the appearance of (137)Cs. In contrast, the highest levels in WB-2 occurred well before the appearance of (137)Cs. Although stricter environmental regulations have caused a decrease in metal concentrations since the 1980s, the concentrations in the surface sediments of many trace metals were elevated to levels 2-5 times higher than the levels at the bottom of the cores in both the Southern and Western Branches of the river.

Creating mixed Fe/Al coatings on planar gamma-Al2O3 surfaces.

Insufficient understanding of the interactions of reactive phases (e.g., Fe and Al oxides) with minerals, other reactive phases and sorbing species has made predicting and modeling metal sorption on natural sediment surfaces difficult. This work develops a method to create mixed Fe/Al planar oxide surfaces by coating well-characterized planar gamma-Al2O3 with ferric iron. The objective is to closely control the Fe/Al ratio as well as the distribution of Fe on the planar surface. Effects of starting Fe(III) concentration, reaction time and number of coating sequences were examined using XPS and ToF-SIMS. No observable trend was seen in Fe/Al ratios by varying the starting Fe(III) concentration or reaction time. For both 4- and 14-day reactions, lower concentrations of Fe(III) produced oxide phases with a homogeneous distribution of Fe at the surface as detected by ToF-SIMS. ToF-SIMS Fe elemental maps of the oxide phases resulting from the highest Fe(III) concentration showed areas of localized Fe deposition. A sequential coating procedure allowed for a closer control of the concentration and spatial distribution of Fe(III) in the resulting oxide phase. This work provides methodology that can be used to create Fe/Al oxide phases whose Fe/Al content can be controlled for use in subsequent sorption studies to better understand the effects of mixed phase oxides on metal ion uptake.

Pb(II) sorption onto gamma-Al2O3 surfaces at the oxide-water interface: a novel approach using planar oxides.

A novel technique for examining metal-ion interactions at the solid-water interface is introduced. Planar oxides, flat, thin coatings of uniform thickness created on a metal support, have been constructed as useful analogs for investigating metal-solid interactions under a variety of conditions. XPS and ToF/SIMS results from sorption studies at pH 6.0 show that the sorption behavior of Pb on each phase is similar with Pb binding preferentially to the bulk gamma-Al(2)O(3). This may be due to the presence of defect sites on the bulk oxides, the preferential exposure of a specific crystallographic plane in the planar oxides, or it may be an artifact of instrumental analysis. A second study examining Pb sorption to planar gamma-Al(2)O(3) under a series of increasingly complex conditions shows that our methods are able to successfully characterize sorption complexes formed in the presence of environmentally derived complexants. Results suggest that Pb is more strongly complexed by aqueous phase organic matter than sediment-bound organic material, indicating a possible control on Pb sorption in natural environments. Overall, the use of planar oxides combined with a powerful suite of spectroscopic tools provides a promising approach to better understanding metal ion sorption to natural sediment surfaces in aquatic environments.

Spatial survey of trace metal contaminants in the sediments of the Elizabeth River, Virginia.

The Elizabeth River is a sub-estuary of the James River, the most southern tributary of the Chesapeake Bay. It is a highly industrialized area, and has been designated a "toxic hot spot" due to the heavy loads of contaminant metals and organic compounds in its bed sediments. Fifty surface sediment samples were taken along the channels and shoals of the Mainstem and the Southern Branch portions of the river. The samples were analyzed for trace metals (Cd, Cr, Cu, Pb and Zn), Fe and particle properties (% sand, % silt, % clay and specific surface area) to discern the spatial distribution of contaminant metals in this system. Enrichment factors were calculated to assess the overall level of contamination relative to other surrounding waterways as well as historic contamination levels within the river itself. The highest levels of metals were found to be in close proximity to industrial sources of contamination. The overall level of contamination was 3-10 times higher than baseline levels within the river, and 2-3 times higher than contaminant levels in other area waters.