Resettled Iraqi Refugees in the United States

The American war against Iraq has caused hundreds of thousands of deaths and displaced millions of people. Between 20 March 2003 and 30 September 2017, more than 172,000 Iraqis resettled in the United States. This book explores the experiences of fifteen Iraqis who resettled in the US after 2003. It examines the long war against Iraq that began in 1991 and the decisions some Iraqis made to leave their homes and seek refuge in the United States. The book also delves into the possibilities for belonging and cultural exchange for this cohort of Iraqis and their political engagement with non-profit organizations, advocacy, and activism against the 2017 Travel Ban

The AP-2 adaptor β2 appendage scaffolds alternate cargo endocytosis

The independently folded appendages of the large α and β2 subunits of the endocytic adaptor protein (AP)-2 complex coordinate proper assembly and operation of endocytic components during clathrin-mediated endocytosis. The β2 subunit appendage contains a common binding site for β-arrestin or the autosomal recessive hypercholesterolemia (ARH) protein. To determine the importance of this interaction surface in living cells, we used small interfering RNA-based gene silencing. The effect of extinguishing β2 subunit expression on the internalization of transferrin is considerably weaker than an AP-2 α subunit knockdown. We show the mild sorting defect is due to fortuitous substitution of the β2 chain with the closely related endogenous β1 subunit of the AP-1 adaptor complex. Simultaneous silencing of both β1 and β2 subunit transcripts recapitulates the strong α subunit RNA interference (RNAi) phenotype and results in loss of ARH from endocytic clathrin coats. An RNAi-insensitive β2-yellow fluorescent protein (YFP) expressed in the β1 + β2-silenced background restores cellular AP-2 levels, robust transferrin internalization, and ARH colocalization with cell surface clathrin. The importance of the β appendage platform subdomain over clathrin for precise deposition of ARH at clathrin assembly zones is revealed by a β2-YFP with a disrupted ARH binding interface, which does not restore ARH colocalization with clathrin. We also show a β-arrestin 1 mutant, which engages coated structures in the absence of any G protein-coupled receptor stimulation, colocalizes with β2-YFP and clathrin even in the absence of an operational clathrin binding sequence. These findings argue against ARH and β-arrestin binding to a site upon the β2 appendage platform that is later obstructed by polymerized clathrin. We conclude that ARH and β-arrestin depend on a privileged β2 appendage site for proper cargo recruitment to clathrin bud sites

The Second Transmembrane Domain of P2X7 Contributes to Dilated Pore Formation

Activation of the purinergic receptor P2X7 leads to the cellular permeability of low molecular weight cations. To determine which domains of P2X7 are necessary for this permeability, we exchanged either the C-terminus or portions of the second transmembrane domain (TM2) with those in P2X1 or P2X4. Replacement of the C-terminus of P2X7 with either P2X1 or P2X4 prevented surface expression of the chimeric receptor. Similarly, chimeric P2X7 containing TM2 from P2X1 or P2X4 had reduced surface expression and no permeability to cationic dyes. Exchanging the N-terminal 10 residues or C-terminal 14 residues of the P2X7 TM2 with the corresponding region of P2X1 TM2 partially restored surface expression and limited pore permeability. To further probe TM2 structure, we replaced single residues in P2X7 TM2 with those in P2X1 or P2X4. We identified multiple substitutions that drastically changed pore permeability without altering surface expression. Three substitutions (Q332P, Y336T, and Y343L) individually reduced pore formation as indicated by decreased dye uptake and also reduced membrane blebbing in response to ATP exposure. Three others substitutions, V335T, S342G, and S342A each enhanced dye uptake, membrane blebbing and cell death. Our results demonstrate a critical role for the TM2 domain of P2X7 in receptor function, and provide a structural basis for differences between purinergic receptors. © 2013 Sun et al

Methylthioadenosine reprograms macrophage activation through adenosine receptor stimulation

Regulation of inflammation is necessary to balance sufficient pathogen clearance with excessive tissue damage. Central to regulating inflammation is the switch from a pro-inflammatory pathway to an anti-inflammatory pathway. Macrophages are well-positioned to initiate this switch, and as such are the target of multiple therapeutics. One such potential therapeutic is methylthioadenosine (MTA), which inhibits TNFα production following LPS stimulation. We found that MTA could block TNFα production by multiple TLR ligands. Further, it prevented surface expression of CD69 and CD86 and reduced NF-KB signaling. We then determined that the mechanism of this action by MTA is signaling through adenosine A2 receptors. A2 receptors and TLR receptors synergized to promote an anti-inflammatory phenotype, as MTA enhanced LPS tolerance. In contrast, IL-1β production and processing was not affected by MTA exposure. Taken together, these data demonstrate that MTA reprograms TLR activation pathways via adenosine receptors to promote resolution of inflammation. © 2014 Keyel et al

Ecosystem functions of oil palm plantations - a review

Oil palm plantations have expanded rapidly in the last decades. This large-scale land-use change has had great impacts on both the areas converted to oil palm and their surroundings. Howev-er, research on the impacts of oil palm agriculture is scattered and patchy, and no clear overview ex-ists. Here, we address this gap through a systematic and comprehensive literature review of all ecosys-tem functions in oil palm plantations. We compare ecosystem functions in oil palm plantations to those in forests as forests are often cleared for the establishment of oil palm. We find that oil palm planta-tions generally have reduced ecosystem functioning compared to forests. Some of these functions are lost globally, such as those to gas and climate regulation and to habitat and nursery functions. The most serious impacts occur when land is cleared to establish new plantations, and immediately after-wards, especially on peat soils. To variable degrees, plantation management can prevent or reduce losses of some ecosystem functions. The only ecosystem function which increased in oil palm planta-tions is, unsurprisingly, the production of marketable goods. Our review highlights numerous research gaps. In particular, there are significant gaps with respect to information functions (socio-cultural functions). There is a need for empirical data on the importance of spatial and temporal scales, such as the differences between plantations in different environments, of different sizes, and of different ages. Finally, more research is needed on developing management practices that can off-set the losses of ecosystem functions

Dynamics and Cargo Selectivity of Endocytic Adaptor Proteins

Clathrin-mediated endocytosis is a critical process through which a wide variety of extracellular material is internalized. The primary component, clathrin, forms a cargo-selective lattice at the plasma membrane, as well as on endosomes and the TGN, though the cargo-selective components are incompletely defined. An ideal tool for understanding the spatio-temporal dynamics of both the clathrin coat and the cargo selected is total internal reflection fluorescence microscopy (TIR-FM), which permits selective imaging of events closely apposed to the ventral plasma membrane. Previously, observation of the clathrin coat has shown both static and dynamic populations, with some dynamic structures undergoing microtubule-dependent motion; the 70-110 nm decay constant of the TIR-FM field has led to the assumption that these are all representative of coated pits. Here, I demonstrate that the dynamic population of clathrin is primarily endosomal, as it lacks colocalization with the plasma membrane-specific endocytic adaptor AP-2, but colocalizes with large, internalized low density lipoprotein (LDL) and transferrin positive structures. Other clathrin-associated sorting proteins (CLASPs) remain in relatively static structures as well. One such CLASP, autosomal recessive hypercholesterolemia (ARH) protein, is the defective protein in ARH, which is typified by the failure of hepatic LDL receptor internalization, despite no LDL receptor mutations. ARH interacts with AP-2 via the novel, helical FXX[FL]XXXR motif present in its C-terminus. Here, I demonstrate the importance of this motif for targeting ARH to coated pits in cells and LDL uptake. As knockdown of ARH is insufficient to block LDL receptor endocytosis in fibroblasts, I show that the CLASP Disabled-2 (Dab2) works with ARH to sort the LDL receptor. Ablation of these two components using RNAi halts LDL receptor endocytosis, and either exogenous ARH or Dab2 rescue this phenotype. The endocytic defect in the liver of ARH patients is due to the lack of Dab2 expression in hepatocytes, making this cell type sensitive to ARH levels for LDL uptake. This work formally validates the CLASP hypothesis, and demonstrates that these CLASPs are general components of the clathrin-coated pit that are regulated in a tissue-specific fashion

A clathrin coat assembly role for the muniscin protein central linker revealed by TALEN-mediated gene editing

Clathrin-mediated endocytosis is an evolutionarily ancient membrane transport system regulating cellular receptivity and responsiveness. Plasmalemma clathrin-coated structures range from unitary domed assemblies to expansive planar constructions with internal or flanking invaginated buds. Precisely how these morphologically-distinct coats are formed, and whether all are functionally equivalent for selective cargo internalization is still disputed. We have disrupted the genes encoding a set of early arriving clathrin-coat constituents, FCHO1 and FCHO2, in HeLa cells. Endocytic coats do not disappear in this genetic background; rather clustered planar lattices predominate and endocytosis slows, but does not cease. The central linker of FCHO proteins acts as an allosteric regulator of the prime endocytic adaptor, AP-2. By loading AP-2 onto the plasma membrane, FCHO proteins provide a parallel pathway for AP-2 activation and clathrin-coat fabrication. Further, the steady-state morphology of clathrin-coated structures appears to be a manifestation of the availability of the muniscin linker during lattice polymerization. DOI: http://dx.doi.org/10.7554/eLife.04137.00

Evaluating The Roles Of Visual Openness And Edge Effects On Nest-Site Selection And Reproductive Success In Grassland Birds

In some species, habitat edges (ecotones) affect nest-site selection and nesting success. Openness, or how visually open a habitat is, has recently been shown to influence grassland bird density and may affect nest-site selection, possibly by reducing the risk of predation on adults, nests, or both. Because edge and openness are correlated, it is possible that effects of openness have been overlooked or inappropriately ascribed to edge effects. We tested the roles of edges and visual openness in nest-site selection and nesting success of two grassland passerines, the Bobolink (Dolichonyx oryzivorus) and Savannah Sparrow (Passerculus sandwichensis), in the Champlain Valley, Vermont. We also evaluated the sensitivity of our results to alternative definitions of edge on our landscape. Bobolink (n = 580) and Savannah Sparrow nests (n = 922) were located on seven hay fields and three pastures from 2002 to 2010. Both species avoided placing nests near edges and in less open habitat compared with expectations based on random placement. When the effects of openness and edge were separated, less open habitats were still avoided, but edge responses were less clear. These results were robust to different definitions of habitat edge. We found no strong relationships between either openness or edges and reproductive success (numbers of eggs and fledglings, percentage of eggs producing fledglings, and nest success), although there may be an edge-specific openness effect on timing of reproduction (clutch completion date). Our results support openness as an important factor in nest-site selection by grassland birds

Roles of AP-2 in clathrin-mediated endocytosis.

The notion that AP-2 clathrin adaptor is an essential component of an endocytic clathrin coat appears to conflict with recent observations that substantial AP-2 depletion, using RNA interference with synthesis of AP-2 subunits, fails to block uptake of certain ligands known to internalize through a clathrin-based pathway