Nitrite reduction by xanthine oxidase family enzymes: a new class of nitrite reductases

J Biol Inorg Chem (2011) 16:443–460 DOI 10.1007/s00775-010-0741-zMammalian xanthine oxidase (XO) and Desulfovibrio gigas aldehyde oxidoreductase (AOR) are members of the XO family of mononuclear molybdoenzymes that catalyse the oxidative hydroxylation of a wide range of aldehydes and heterocyclic compounds. Much less known is the XO ability to catalyse the nitrite reduction to nitric oxide radical (NO). To assess the competence of other XO family enzymes to catalyse the nitrite reduction and to shed some light onto the molecular mechanism of this reaction, we characterised the anaerobic XO- and AORcatalysed nitrite reduction. The identification of NO as the reaction product was done with a NO-selective electrode and by electron paramagnetic resonance (EPR) spectroscopy. The steady-state kinetic characterisation corroborated the XO-catalysed nitrite reduction and demonstrated, for the first time, that the prokaryotic AOR does catalyse the nitrite reduction to NO, in the presence of any electron donor to the enzyme, substrate (aldehyde) or not (dithionite). Nitrite binding and reduction was shown by EPR spectroscopy to occur on a reduced molybdenum centre. A molecular mechanism of AOR- and XO-catalysed nitrite reduction is discussed, in which the higher oxidation states of molybdenum seem to be involved in oxygen-atom insertion, whereas the lower oxidation states would favour oxygenatom abstraction. Our results define a new catalytic performance for AOR—the nitrite reduction—and propose a new class of molybdenum-containing nitrite reductases. Keywords Nitrite reduction Nitric oxide formation Molybdenum Xanthine oxidase Aldehyde oxidoreductase Abbreviations AOR Aldehyde oxidoreductase DMSOR Dimethylsulfoxide reductase EPR Electron paramagnetic resonance Fe/S Iron–sulfur centre Fe/S–NO Dinitrosyl–iron–sulfur complex (MGD)2–Fe Ferrous complex of di(N-methyl-Dglucamine dithiocarbamate)(MGD)2–Fe–NO Mononitrosyl–iron complex Mo-enzymes Pterin–molybdenum-containing enzymes NaR Nitrate reductases NO Nitric oxide radical SO Sulfite oxidase XO Xanthine oxidase Introduction Molybdenum is present in a wide variety of enzymes, in both prokaryotes and eukaryotes, where it performs catalyti

What smells? Developing in-field methods to characterize the chemical composition of wild mammalian scent cues

Olfactory cues play an important role in mammalian biology, but have been challenging to assess in the field. Current methods pose problematic issues with sample storage and transportation, limiting our ability to connect chemical variation in scents with relevant ecological and behavioral contexts. Real-time, in-field analysis via portable gas chromatography–mass spectrometry (GC-MS) has the potential to overcome these issues, but with trade-offs of reduced sensitivity and compound mass range. We field-tested the ability of portable GC-MS to support two representative applications of chemical ecology research with a wild arboreal primate, common marmoset monkeys (Callithrix jacchus). We developed methods to (a) evaluate the chemical composition of marmoset scent marks deposited at feeding sites and (b) characterize the scent profiles of exudates eaten by marmosets. We successfully collected marmoset scent marks across several canopy heights, with the portable GC-MS detecting known components of marmoset glandular secretions and differentiating these from in-field controls. Likewise, variation in the chemical profile of scent marks demonstrated a significant correlation with marmoset feeding behavior, indicating these scents’ biological relevance. The portable GC-MS also delineated species-specific olfactory signatures of exudates fed on by marmosets. Despite the trade-offs, portable GC-MS represents a viable option for characterizing olfactory compounds used by wild mammals, yielding biologically relevant data. While the decision to adopt portable GC-MS will likely depend on site- and project-specific needs, our ability to conduct two example applications under relatively challenging field conditions bodes well for the versatility of in-field GC-MS

Energy metabolism in human pluripotent stem cells and their differentiated counterparts

Background: Human pluripotent stem cells have the ability to generate all cell types present in the adult organism, therefore harboring great potential for the in vitro study of differentiation and for the development of cell-based therapies. Nonetheless their use may prove challenging as incomplete differentiation of these cells might lead to tumoregenicity. Interestingly, many cancer types have been reported to display metabolic modifications with features that might be similar to stem cells. Understanding the metabolic properties of human pluripotent stem cells when compared to their differentiated counterparts can thus be of crucial importance. Furthermore recent data has stressed distinct features of different human pluripotent cells lines, namely when comparing embryo-derived human embryonic stem cells (hESCs) and induced pluripotent stem cells (IPSCs) reprogrammed from somatic cells. Methodology/Principal Findings: We compared the energy metabolism of hESCs, IPSCs, and their somatic counterparts. Focusing on mitochondria, we tracked organelle localization and morphology. Furthermore we performed gene expression analysis of several pathways related to the glucose metabolism, including glycolysis, the pentose phosphate pathway and the tricarboxylic acid (TCA) cycle. In addition we determined oxygen consumption rates (OCR) using a metabolic extracellular flux analyzer, as well as total intracellular ATP levels by high performance liquid chromatography (HPLC). Finally we explored the expression of key proteins involved in the regulation of glucose metabolism. Conclusions/Findings: Our results demonstrate that, although the metabolic signature of IPSCs is not identical to that of hESCs, nonetheless they cluster with hESCs rather than with their somatic counterparts. ATP levels, lactate production and OCR revealed that human pluripotent cells rely mostly on glycolysis to meet their energy demands. Furthermore, our work points to some of the strategies which human pluripotent stem cells may use to maintain high glycolytic rates, such as high levels of hexokinase II and inactive pyruvate dehydrogenase (PDH). © 2011 Varum et al

Spatial eigensolution analysis of discontinuous Galerkin schemes with practical insights for under-resolved computations and implicit LES

The study focusses on the dispersion and diffusion characteristics of discontinuous spectral element methods - specifically discontinuous Galerkin (DG) - via the spatial eigensolution analysis framework built around a one-dimensional linear problem, namely the linear advection equation. Dispersion and diffusion characteristics are of critical importance when dealing with under-resolved computations, as they affect both the numerical stability of the simulation and the solution accuracy. The spatial eigensolution analysis carried out in this paper complements previous analyses based on the temporal approach, which are more commonly found in the literature. While the latter assumes periodic boundary conditions, the spatial approach assumes inflow/outflow type boundary conditions and is therefore better suited for the investigation of open flows typical of aerodynamic problems, including transitional and fully turbulent flows and aeroacoustics. The influence of spurious/reflected eigenmodes is assessed with regard to the presence of upwind dissipation, naturally present in DG methods. This provides insights into the accuracy and robustness of these schemes for under-resolved computations, including under-resolved direct numerical simulation (uDNS) and implicit large-eddy simulation (iLES). The results estimated from the spatial eigensolution analysis are verified using the one-dimensional linear advection equation and successively by performing two-dimensional compressible Euler simulations that mimic (spatially developing) grid turbulence

Berry connections calculated from first principles

Berry connections can be very useful for the determination of electronic properties of materials. Yet they are particularly difficult to calculate from first principles. The main problems in so calculating are the randomness of the wavefunction phase for different points in the reciprocal space that is given by numerical calculations and the band crossing at points of degeneracy. In this letter we show how this can be overcome and applied to the simple case of two dimensional semiconductor hexagonal boron nitride. To demonstrate the effectiveness of the process, we calculate the linear and second harmonic generation conductivities from the Berry connections obtained directly from density functional theory

Direct electrochemical reduction of carbon dioxide by a molybdenum-containing formate dehydrogenase

This work was supported by the Associate Laboratory for Green Chemistry -LAQV, with national funds from FCT/MCTES ( UID/QUI/50006/2019 ). LBM thanks to FCT/MCTES for the CEEC-Individual 2017 Program Contract.Formate dehydrogenase enzymes catalyse the reversible two-electron oxidation of formate to carbon dioxide. The class of metal-dependent formate dehydrogenases comprises prokaryotic enzymes holding redox-active centres and a catalytic site, containing either molybdenum or tungsten ion, that mediates the formate/carbon dioxide interconversion. The carbon dioxide reduction is of a particular interest, since it may be a route for its atmospheric mitigation with the simultaneous production of added-value products, as formate-derived compounds. Recently, the periplasmic formate dehydrogenase from Desulfovibrio desulfuricans, a molybdenum-containing enzyme, was proven to be an efficient enzyme for the CO 2 reduction to formate. In this work, the immobilized formate dehydrogenase isolated from Desulfovibrio desulfuricans direct electrochemical behaviour was attained in the presence and absence of substrates and the formal potentials associated with the catalytic centre transitions were determined in non-turnover conditions. The enzyme catalytic activity towards carbon dioxide reduction was observed using direct electrochemical methods.authorsversionpublishe

Laboratório de enologia.

A vitivinicultura no Submédio São Francisco.bitstream/item/132811/1/ID-42731.pd

Tendências fluviométricas nas áreas estuarinas de Goiana-Megaó e Pirapama/Jaboatão e das tabuas de maré no Porto de Suape-PE.

Os estuários são ambientes de transição entre o oceano e o continente, ocorrendo na desmbocadura dos rios, resultando na diluição da água salgada

Análise da precipitação e do número de dias de chuva em Juazeiro, Bahia.

O objetivo deste trabalho foi analisaro comportamento da precipitação e do número dedias com chuva no município de Juazeiro ? BA,tendo em vista fornecer subsídios auxiliares aoplanejamento agropecuário durante o ano