The Roles of Membrane Rafts in CD32A-Mediated Phagocytosis

Membrane rafts are highly dynamic heterogeneous sterol- and sphingolipid-rich micro-domains on cell surfaces. They are generally believed to provide residency for cell surface molecules (e.g., adhesion and signaling molecules) and scaffolding to facilitate the functions of these molecules such as membrane trafficking, receptor transport, cell signaling, and endocytosis.
The governing, or overall hypothesis, for this project is that membrane rafts provide residency for Fc[gamma]RIIA (CD32A) on K562 cells, and that by doing so they provide a platform from which Fc[gamma]RIIA initiate or carry out their functions, which include migration, signaling, phagocytic synapse formation, and internalization of IgG opsonized targets.
Using immuno-fluorescent laser scanning confocal microscopy and reflection interference microscopy (RIM), we studied the spatial and temporal distributions of membrane rafts and surface receptors, signaling molecules, and cell organelles during the formation of phagocytic contact areas. K562 cells, which naturally express CD32A, a cell surface receptor for the Fc portion of Immuno-globulin G(IgG), was chosen as a model for neutrophils. An opsonized target was modeled using a glass supported lipid bilayer reconstituted with IgG. CD32A was found to cluster and co-localize with membrane rafts. Placing the K562 cells on the lipid bilayer triggered a process of contact area formation that includes binding between receptors and ligands, their recruitment to the contact area, a concurrent membrane raft movement to and concentration in the contact area, and transport of CD32A, IgG, and membrane rafts to the Golgi complex. Characterization of these processes was performed using agents known to disrupt detergent resistant membranes (DRMs), dissolve actin microfilaments, and inhibit myosin motor activity, which abolished the CD32A clusters and prevented the contact area formation. 
The relevance to phagocytosis of contact area formation between K562 cells and lipid bilayers was demonstrated using micro-beads coated with a lipid bilayer reconstituted with IgG as the opsonized target instead of the glass supported planar lipid bilayer. Disruption of membrane rafts, salvation of the actin cytoskeleton, and inhibition of myosin II activity were found to inhibit phagocytosis.
These data suggest membrane rafts play several important roles in CD32A mediated phagocytosis including pre-clustering CD32A, transport of CD32A to the phagocytic cup, and transport of the opsonized target towards the Golgi complex. Here we have provided evidence that membrane rafts serve as platforms which are used to cluster CD32A and transport CD32A along the actin cytoskeleton to the site of phagocytic synapse formation thus allowing for the quick assembly of a phagocytic synapse.
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Iron: the forgotten driver of nitrous oxide production in agricultural soil.

In response to rising interest over the years, many experiments and several models have been devised to understand emission of nitrous oxide (N2O) from agricultural soils. Notably absent from almost all of this discussion is iron, even though its role in both chemical and biochemical reactions that generate N2O was recognized well before research on N2O emission began to accelerate. We revisited iron by exploring its importance alongside other soil properties commonly believed to control N2O production in agricultural systems. A set of soils from California's main agricultural regions was used to observe N2O emission under conditions representative of typical field scenarios. Results of multivariate analysis showed that in five of the twelve different conditions studied, iron ranked higher than any other intrinsic soil property in explaining observed emissions across soils. Upcoming studies stand to gain valuable information by considering iron among the drivers of N2O emission, expanding the current framework to include coupling between biotic and abiotic reactions

Complete integrin headpiece opening in eight steps

Carefully soaking crystals with Arg-Gly-Asp (RGD) peptides, we captured eight distinct RGD-bound conformations of the αIIbβ3 integrin headpiece. Starting from the closed βI domain conformation, we saw six intermediate βI conformations and finally the fully open βI with the hybrid domain swung out in the crystal lattice. The β1-α1 backbone that hydrogen bonds to the Asp side chain of RGD was the first element to move followed by adjacent to metal ion-dependent adhesion site Ca2+, α1 helix, α1’ helix, β6-α7 loop, α7 helix, and hybrid domain. We define in atomic detail how conformational change was transmitted over long distances in integrins, 40 Å from the ligand binding site to the opposite end of the βI domain and 80 Å to the far end of the hybrid domain. During these movements, RGD slid in its binding groove toward αIIb, and its Arg side chain became ordered. RGD concentration requirements in soaking suggested a >200-fold higher affinity after opening. The thermodynamic cycle shows how higher affinity pays the energetic cost of opening

Widespread dispersal and aging of organic carbon in shallow marginal seas

The occurrence of pre-aged organic carbon (OC) in continental margin surface sediments is a commonly observed phenomenon, yet the nature, sources, and causes of this aged OC remain largely undetermined for many continental shelf settings. Here we present the results of an extensive survey of the abundance and radiocarbon content of OC in surface sediments from the northern Chinese marginal seas. Pre-aged OC is associated with both coarser (>63 µm) and finer (<63 µm) sedimentary components; measurements on specific grain-size fractions reveal that it is especially prevalent within the 20–63 µm fraction of inner shelf sediments. We suggest that organic matter associated with this sortable silt fraction is subject to protracted entrainment in resuspension-deposition loops during which it ages, is modified, and is laterally dispersed, most likely via entrainment within benthic nepheloid layers. This finding highlights the complex dynamics and predepositional history of organic matter accumulating in continental shelf sediments, with implications for our understanding of carbon cycling on continental shelves, development of regional carbon budgets, and interpretation of sedimentary records

Interaction of a symmetrical α,α',δ,δ'-Tetramethyl-cucurbit[6]uril with Ln³⁺ : potential applications for isolation of lanthanides

The interaction of a symmetrical α,α′,δ,δ′-tetramethyl-cucurbit[6]uril (TMeQ[6]) with a series of lanthanide cations (Ln³⁺) was investigated in neutral water and in acidic solution. Analysis by single crystal X-ray diffraction revealed that different isomorphous families formed under different synthetic conditions. Such differences in the interaction between TMeQ[6] and Ln³⁺ could potentially be used for isolating heavier Ln³⁺ from their lighter counterparts in neutral solution, and lighter lanthanide cations from their heavier counterparts in acidic solution

Supramolecular assembly of cucurbit[6]uril and N-butyl-4-pyrrolidinopyridine

The nature of the supramolecular host-guest complex involving 4-pyrrolidinopyridine (BuPC4) and cucurbit[6]uril (Q[6]) has been investigated by NMR and UV spectroscopy, MALDI-TOF mass spectrometry, X-ray crystallography and isothermal titration calorimetry (ITC). The results revealed that the alkyl chain of the guest BuPC4 is located inside the cavity of the Q[6] host, whereas the other section of the BuPC4 guest remains outside of the portal

Green and blue water accounting in the Limpopo and Nile Basins: Implications for food and agricultural policy

Green water, Blue water, Irrigation, Rainfed, Agriculture, technology, Investment, Impact, Climate change, Water resources,