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<p>Uche Amazigo</p

Butchered meat following a communal hunt.

<p>A cane rat (<i>Thryonomys swinderianus</i>) caught during a communal hunt. The carcass is singed, butchered and shared between the participants who either cook it together or bring it back to their household.</p

Communal hunting with nets.

<p>A cane rat (<i>Thryonomys swinderianus</i>) caught during a communal hunt. The animal was bludgeoned to death by hitting the skull with bare fists, rather than a machete, to avoid damaging the hunting net.</p

Children hunting.

<p>Children hunting with dogs owned by the family and borrowed from an unknowing neighbour (A). The dogs detect or chase rodents into burrows, which are then dug up by the children. A genet kitten (<i>Genetta</i> sp.) (B) The kittens were reared by children in a chicken coop and eventually eaten.</p

Snare traps.

<p>A common trap (<i>dahin</i>), which can catch most species of mammals and reptiles (A) and a trap specifically designed to snare small non-human primates (B) as they cross a cleared portion of forest on a branch. These latter traps are uncommon because they are difficult to build and non-human primates learn to avoid them. Both traps work by snaring animals with use of a spring mechanisms when they pass through a sensitive trigger mechanism (arrows).</p

Development of Bright and Biocompatible Nanoruby and Its Application to Background-Free Time-Gated Imaging of G‑Protein-Coupled Receptors

At the forefront of developing fluorescent probes for biological imaging applications are enhancements aimed at increasing their brightness, contrast, and photostability, especially toward demanding applications of single-molecule detection. In comparison with existing probes, nanorubies exhibit unlimited photostability and a long emission lifetime (∼4 ms), which enable continuous imaging at single-particle sensitivity in highly scattering and fluorescent biological specimens. However, their wide application as fluorescence probes has so far been hindered by the absence of facile methods for scaled-up high-volume production and molecularly specific targeting. The present work encompasses the large-scale production of colloidally stable nanoruby particles, the demonstration of their biofunctionality and negligible cytotoxicity, as well as the validation of its use for targeted biomolecular imaging. In addition, optical characteristics of nanorubies are found to be comparable or superior to those of state-of-the-art quantum dots. Protocols of reproducible and robust coupling of functional proteins to the nanoruby surface are also presented. As an example, NeutrAvidin-coupled nanoruby show excellent affinity and specificity to μ-opioid receptors in fixed and live cells, allowing wide-field imaging of G-protein coupled receptors with single-particle sensitivity

Inhibition of anterograde FAT induced by mSOD1 depends on specific MKKK-MKK interactions.

<p>Co-perfusion of G93A-SOD1 with DVD peptide <b>(a)</b>, but not with the Mixed-Lineage Kinase inhibitor CEP11004 <b>(b),</b> prevents inhibition of FAT induced by G93A-SOD1. DVD peptide prevents activation of MKKs by some MKKKs (n =  number of axoplasms) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0065235#pone.0065235-Takekawa1" target="_blank">[60]</a>. These data suggest that the activation of p38 and the inhibition of FAT induced by G93A-SOD1 involves activation of one or more MAPKKKs <i>other</i> than MLKs. <b>(c)</b> The DVD peptide also blocks inhibition of FAT by oxidized WT-SOD1 suggesting that FALS mutant SOD1 and misfolded WT-SOD1 activate a common p38 MAPK pathway <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0065235#pone.0065235-Bosco2" target="_blank">[23]</a>.</p

Pathogenic SOD1 increases neurofilament phosphorylation.

<p>Phosphorylation of squid neurofilaments (NF) in isolated “sister” axoplasms (see Methods) was analyzed using metabolic labeling experiments with ³²P-γ-ATP. (<b>a)</b> Coomassie Blue staining (CB) shows similar levels of perfused WT-SOD1, G93A-SOD1 and total axoplasmic proteins. Immunoblot analysis (WB) with the NFH antibody SMI-31 confirmed the identity of major phosphorylated bands as NF220 and HMW, major NF subunits in squid axoplasm <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0065235#pone.0065235-Pant1" target="_blank">[30]</a>. Short (S) and long (L) exposure of autoradiograms (³²P) show increased phosphorylation of NF220 and HMW NF subunits in axoplasms perfused with G93A-SOD1, compared to WT-SOD1. <b>(b)</b> Quantitation of squid NF phosphorylation showed ⋍70% increase in G93A-SOD1 treated axoplasms, compared to those treated with WT-SOD1 (p≤0.01 (#) in a paired t-test). <b>(c)</b> In parallel experiments, kinesin-1 was immunoprecipitated from axoplasms labeled with γ-³²P-ATP in the presence of WT-SOD1 or G93A-SOD1. Both heavy (KHC) and light (KLC) chains of conventional kinesin were phosphorylated. <b>(d)</b> The ratio of counts for G93A-SOD1/WT-SOD1 indicates that KHC labeling increased 31% in G93A-SOD1 axoplasms, compared to WT-SOD1 (significant at p≤0.05 by paired t-test, #). KLC phosphorylation increased by 15%, but was not statistically significant (p = 0.123). n = 7.</p

Pseudophosphorylation of kinesin-1 at S175/S176 inhibits movement of kinesin-1.

<p>To determine the effects of modifying S175 and S176 on kinesin-1function, recombinant GFP-tagged kinesin (KHC⁵⁵⁹) was modified to preclude phosphorylation at these sites (S175AS176A) or to mimic phosphorylation (S175ES176E). <b>(a–f)</b> Stage 3 hippocampal neurons were examined 5 h after co-transfection with GFP-tagged KHC⁵⁵⁹ constructs and a tdTomato construct, which diffuses throughout the cell and allows visualization of neurites (<b>b, d, f</b>). Both wild-type kinesin-1 (KHC⁵⁵⁹ WT, <b>a</b>) and a non-phosphorylatable mutant (KHC⁵⁵⁹ S175A/S176A, <b>c</b>) accumulated efficiently at axonal tips (labeled by arrows) with minimal steady-state labeling of cell bodies (arrowheads). In contrast, pseudophosphorylated mutant KHC⁵⁵⁹ S175E/S176E, <b>e</b>) was mainly present in neuronal cell bodies. Quantitative immunofluorescence analysis shows fraction of total KHC⁵⁵⁹ fluorescence at axon tips for all constructs <b>(g)</b>. Far less phosphomimicking KHC⁵⁵⁹ S175E/S176E constructs accumulated at axon tips than KHC⁵⁵⁹ WT or KHC⁵⁵⁹ S175A/S176A (#: p<0.001; <i>n</i>: 27–43 cells per condition). Bars show mean and standard deviation. Scale bar  = 20 µm.</p

Active p38 α mimics the effects of pathogenic SOD1 on anterograde FAT.

<p>Effects of active, recombinant p38 isoforms on FAT were evaluated using vesicle motility assays in isolated squid axoplasm. P38α and P38β were perfused at a constant specific activity based on <i>in vitro</i> kinase assays with the ATF-2 substrate. <b>(a)</b> Perfusion of active p38α in axoplasm selectively inhibited anterograde FAT, as did pathogenic SOD1 (compare to Fig. 1b-d). (<b>b)</b> Unlike p38α, p38β inhibited both anterograde and retrograde FAT. (<b>c</b>) Quantitation of values obtained between 30-50 minutes shows that p38α most closely mimicked effects of pathogenic SOD1, suggesting this isoform mediates the effects of mSOD1 on FAT in axoplasm (#: difference significant from WT-SOD1 at p<0.01 by t-test).</p