V-ATPase inhibition prevents recovery from anoxia in Artemia franciscana embryos: Quiescence signaling through dissipation of proton gradients

The metabolic downregulation critical for long-term survival of Artemia franciscana embryos under anoxia is mediated, in part, by a progressive intracellular acidification. However, very little is known about the mechanisms responsible for the pH transitions associated with exposure to, and recovery from, oxygen deprivation. In the present study, we demonstrate with 31P-NMR that incubation of intact embryos with the V-ATPase inhibitor bafilomycin A1 severely limits intracellular alkalinization during recovery from anoxia without affecting the restoration of cellular nucleotide triphosphate levels. Based on these data, it appears that oxidative phosphorylation and ATP resynthesis can only account for the first 0.3 pH unit alkalinization observed during aerobic recovery from the 1 pH unit acidification produced during 1 h of anoxia. The additional 0.7 pH unit increase requires proton pumping by the V-ATPase. Aerobic incubation with bafilomycin also suggests that V-ATPase inhibition alone is not enough to induce an acute dissipation of proton gradients under anoxia. In intact embryos, the dissipation of proton gradients and uncoupling of oxidative phosphorylation with carbonyl cyanide 3-chlorophenylhydrazone (CCCP) leads to an intracellular acidification similar to that seen after 1 h of anoxia. Subsequent exposure to anoxia, in the continued presence of CCCP, yields little additional acidification, suggesting that proton gradients are normally dissipated under anoxia. When combined with protons generated from net ATP hydrolysis, these data show that the dissipation of proton chemical gradients is sufficient to account for the reversible acidification associated with quiescence in these embryos

Monitoring the effect of three humic acids on a model membrane system using \u3csup\u3e31\u3c/sup\u3eP NMR

The sorption of three humic acids to 1,2-dipalmitoyl-sn-glycero-3- phosphocholine multilamellar vesicle model membrane systems was studied by phosphorus nuclear magnetic resonance (31P NMR). The effects of temperature and pH were investigated. The gel → bilayer transition did not appear to be affected by any of the humic acids at pH 7; however, all three humic acids induced a perturbation to this transition and to the bilayer structure at pH 4. On the basis of the findings from this and other work, a conceptual adsorption/absorption model for the sorption of humic acid (HA) to biomembranes has been put forward. The model requires an initial adsorption step initiated at an acidic pH by hydrogen bridging and electrostatic interactions between the functional groups of the HAs and the head groups of the phospholipids. Once the HA material is adsorbed, its hydrophobic domains can further seek a more thermodynamically favorable environment within the bilayer using hydrophobic interactions. These interactions lead to the HA being absorbed into the membrane, which subsequently induces the observed perturbation by disturbing the ordered packing of the phospholipid tail groups. This model is also related to other humic substances/biomembrane observations in the literature. © 2008 American Chemical Society

Mild colorimetric detection of sialic acid

As part of a broader program focusing on the detection of colorless biomolecules in the visible region, a simple and mild procedure for the visual detection of the most abundant sialic acid at room temperature and neutral pH is reported. The role of the interaction of the sialic acid amide with boron in a readily synthesized boronic acid-based receptor is an important feature of the signaling mechanism. Selectivity of reaction can be tuned by the judicious choice of solvents. This study embodies a departure from many of the author\u27s earlier efforts in sugar solutions that are not heated, affording relatively more selective and milder detection

Chromophore formation in resorcinarene solutions and the visual detection of mono- and oligosaccharides

The colorimetric properties of resorcinarene solutions had not been investigated since Baeyer\u27s initial synthesis. We recently reported that solutions containing resorcinarene macrocycles develop color upon heating or standing. In the presence of saccharides, these solutions exhibit significant color changes which are easily seen. We herein present strong evidence that the solution color is due to macrocycle ring opening and oxidation. The optical responses to saccharides are due to complexation of the sugar with the acyclic chromophores. We apply these mechanistic insights toward the challenging problem of the visual detection of neutral oligosaccharides by simple chromogens. In addition, we also report the first single-crystal X-ray crystal structure determination of a rarely observed diamond resorcinarene stereoisomer