29,971 research outputs found

    Nitrogen Oxyanion-dependent Dissociation of a Two-component Complex That Regulates Bacterial Nitrate Assimilation

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    Nitrogen is an essential nutrient for growth and is readily available to microbes in many environments in the form of ammonium and nitrate. Both ions are of environmental significance due to sustained use of inorganic fertilizers on agricultural soils. Diverse species of bacteria that have an assimilatory nitrate/nitrite reductase system (NAS) can use nitrate or nitrite as the sole nitrogen source for growth when ammonium is limited. In Paracoccus denitrificans, the pathway-specific two-component regulator for NAS expression is encoded by the nasT and nasS genes. Here, we show that the putative RNA-binding protein NasT is a positive regulator essential for expression of the nas gene cluster (i.e. nasABGHC). By contrast, a nitrogen oxyanion-binding sensor (NasS) is required for nitrate/nitrite-responsive control of nas gene expression. The NasS and NasT proteins co-purify as a stable heterotetrameric regulatory complex, NasS-NasT. This protein-protein interaction is sensitive to nitrate and nitrite, which cause dissociation of the NasS-NasT complex into monomeric NasS and an oligomeric form of NasT. NasT has been shown to bind the leader RNA for nasA. Thus, upon liberation from the complex, the positive regulator NasT is free to up-regulate nas gene expression

    Quark-gluon vertex with an off-shell O(a)-improved chiral fermion action

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    We perform a study the quark-gluon vertex function with a quenched Wilson gauge action and a variety of fermion actions. These include the domain wall fermion action (with exponentially accurate chiral symmetry) and the Wilson clover action both with the non-perturbatively improved clover coefficient as well as with a number of different values for this coefficient. We find that the domain wall vertex function behaves very well in the large momentum transfer region. The off-shell vertex function for the on-shell improved clover class of actions does not behave as well as the domain wall case and, surprisingly, shows only a weak dependence on the clover coefficient cSWc_{SW} for all components of its Dirac decomposition and across all momenta. Including off-shell improvement rotations for the clover fields can make this action yield results consistent with those from the domain wall approach, as well as helping to determine the off-shell improved coefficient cqc_q^\prime.Comment: 11 pages, 13 figures, REVTeX

    Neogenin-mediated hemojuvelin shedding occurs after hemojuvelin traffics to the plamsa membrane

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    Hemochromatosis type 2 gene (HFE2) is highly expressed in skeletal muscle and liver hepatocytes. Its encoded protein, hemojuvelin (HJV), is a co-receptor for the bone morphogenetic proteins 2 and 4 (BMP2 and BMP4) and enhances the BMP-induced hepcidin expression. Hepcidin is a central iron regulatory hormone predominantly secreted from hepatocytes. HJV also binds neogenin, a membrane protein widely expressed in many tissues. Neogenin is required for the processing and release of HJV from cells. The role that neogenin plays in HJV trafficking was investigated, using HepG2 cells, a human hepatoma cell line. Knockdown of endogenous neogenin markedly suppresses HJV release, but has no evident effect on HJV trafficking to the plasma membrane. Addition of a soluble neogenin ectodomain to cells markedly inhibits HJV release, indicating that the HJV shedding is not processed before trafficking to the cell surface. At the plasma membrane it undergoes endocytosis in a dynamin-independent but cholesterol-dependent manner. The additional findings that HJV release is coupled to lysosomal degradation of neogenin, that HJV undergoes endocytosis, and that cholesterol depletion by filipin blocks both HJV endocytosis and HJV release, suggest that neogenin-mediated HJV release occurs after the HJV-neogenin complex is internalized from the cell surface

    Estimation of subsurface porosities and thermal conductivities of polygonal tundra by coupled inversion of electrical resistivity, temperature, and moisture content data

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    Studies indicate greenhouse gas emissions following permafrost thaw will amplify current rates of atmospheric warming, a process referred to as the permafrost carbon feedback. However, large uncertainties exist regarding the timing and magnitude of the permafrost carbon feedback, in part due to uncertainties associated with subsurface permafrost parameterization and structure. Development of robust parameter estimation methods for permafrost-rich soils is becoming urgent under accelerated warming of the Arctic. Improved parameterization of the subsurface properties in land system models would lead to improved predictions and a reduction of modeling uncertainty. In this work we set the groundwork for future parameter estimation (PE) studies by developing and evaluating a joint PE algorithm that estimates soil porosities and thermal conductivities from time series of soil temperature and moisture measurements and discrete in-time electrical resistivity measurements. The algorithm utilizes the Model-Independent Parameter Estimation and Uncertainty Analysis toolbox and coupled hydrological-thermal-geophysical modeling. We test the PE algorithm against synthetic data, providing a proof of concept for the approach. We use specified subsurface porosities and thermal conductivities and coupled models to set up a synthetic state, perturb the parameters, and then verify that our PE method is able to recover the parameters and synthetic state. To evaluate the accuracy and robustness of the approach we perform multiple tests for a perturbed set of initial starting parameter combinations. In addition, we varied types and quantities of data to better understand the optimal dataset needed to improve the PE method. The results of the PE tests suggest that using multiple types of data improve the overall robustness of the method. Our numerical experiments indicate that special care needs to be taken during the field experiment setup so that (1) the vertical distance between adjacent measurement sensors allows the signal variability in space to be resolved and (2) the longer time interval between resistivity snapshots allows signal variability in time to be resolved

    Measuring the electrical impedance of mouse brain tissue

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    We report on an experimental method to measure conductivity of cortical tissue. We use a pair of 5mm diameter Ag/AgCl electrodes in a Perspex sandwich device that can be brought to a distance of 400 microns apart. The apparatus is brought to uniform temperature before use. Electrical impedance of a sample is measured across the frequency range 20 Hz-2.0 MHz with an Agilent 4980A four-point impedance monitor in a shielded room. The equipment has been used to measure the conductivity of mature mouse brain cortex in vitro. Slices 400 microns in thickness are prepared on a vibratome. Slices are bathed in artificial cerebrospinal fluid (ACSF) to keep them alive. Slices are removed from the ACSF and sections of cortical tissue approximately 2 mm times 2 mm are cut with a razor blade. The sections are photographed through a calibrated microscope to allow identification of their cross-sectional areas. Excess ACSF is removed from the sample and the sections places between the electrodes. The impedance is measured across the frequency range and electrical conductivity calculated. Results show two regions of dispersion. A low frequency region is evident below approximately 10 kHz, and a high frequency dispersion above this. Results at the higher frequencies show a good fit to the Cole-Cole model of impedance of biological tissue; this model consists of resistive and non-linear capacitive elements. Physically, these elements are likely to arise due to membrane polarization and migration of ions both intra- and extra-cellularly.http://www.iupab2014.org/assets/IUPAB/NewFolder/iupab-abstracts.pd

    A kinetic and theoretical study of the borate catalysed reactions of hydrogen peroxide: the role of dioxaborirane as the catalytic intermediate for a wide range of substrates

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    Our recent work has provided new insights into the equilibria and species that exist in aqueous solution at different pHs for the boric acid – hydrogen peroxide system, and the role of these species in oxidation reactions. Most recently, (M. C. Durrant, D. M. Davies and M. E. Deary, Org. Biomol. Chem., 2011, 9,7249–7254), we have produced strong theoretical and experimental evidence for the existence of a previously unreported monocyclic three membered peroxide species, dioxaborirane, that is the likely catalytic species in borate mediated electrophilic reactions of hydrogen peroxide in alkaline solution. In the present paper, we extend our study of the borate–peroxide system to look at a wide range of substrates that include substituted dimethyl anilines, methyl-p-tolyl sulfoxide, halides, hydrogen sulfide anion, thiosulfate ,thiocyanate, and hydrazine. The unusual selectivity–reactivity pattern of borate catalysed reactions compared with hydrogen peroxide and inorganic or organic peracids previously observed for theorganic sulfides (D. M. Davies, M. E. Deary, K. Quill and R. A. Smith, Chem.–Eur. J., 2005, 11, 3552–3558) is also seen with substituted dimethyl aniline nucleophiles. This provides evidence that the pattern is not due to any latent electrophilic tendency of the organic sulfides and further supports dioxaborirane being the likely reactive intermediate, thus broadening the applicability of this catalytic system. Moreover, density functional theory calculations on our proposed mechanism involving dioxaborirane are consistent with the experimental results for these substrates. Results obtained at high concentrations of both borate and hydrogen peroxide require the inclusion the diperoxodiborate dianion in the kinetic analysis .A scheme detailing our current understanding of the borate–peroxide system is presented
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