2,888 research outputs found

    Modulation of Queuine Uptake and Incorporation into tRNA by Protein Kinase C and Protein Phosphatase

    Get PDF
    It has been suggested that the rate of queuine uptake into cultured human fibroblasts is controlled by phosphorylation levels within the cell. We show that the uptake of queuine is stimulated by activators of protein kinase C (PKC) and inhibitors of protein phosphatase; while inhibitors of PKC, and down-regulation of PKC by chronic exposure to phorbol esters inhibit the uptake of queuine into cultured human fibroblasts. Activators of cAMP- and cGMP-dependent kinases exert no effect on the uptake of queuine into fibroblast cell cultures. These studies suggest that PKC directly supports the activity of the queuine uptake mechanism, and that protein phosphatase activity in the cell acts to reverse this. Regardless of the modulation of uptake rate, the level of intracellular queuine base saturates in 6 h. However, there is still an effect on the incorporation rate of queuine into tRNA of fibroblast cultures even after 24 h. We now show that the incorporation of queuine into tRNA in cultured human fibroblasts by tRNA-guanine ribosyltransferase (TGRase) is also stimulated by activators of PKC and inhibitors of protein phosphatase; while inhibitors of PKC decrease the activity of this enzyme. These studies suggest that PKC supports both the cellular transport of queuine and the activity of TGRase in cultured human fibroblasts, and that protein phosphatase activity in fibroblasts acts to reverse this phenomenon. A kinase-phosphatase control system, that is common to controlling both intracellular signal transduction and many enzyme systems, appears to be controlling the availability of the queuine substrate and the mechanism for its incorporation into tRNA. Since hypomodification of transfer RNA with queuine is commonly observed in undifferentiated, rapidly growing and neoplastically transformed cells, phosphorylation of the queuine modification system may be a critical regulatory mechanism for the modification of tRNA and subsequent control of cell growth and differentiation

    Activation of Transfer RNA-Guanine Ribosyltransferase by Protein Kinase C

    Get PDF
    Transfer RNA-guanine ribosyltransferase (TGRase) irreversibly incorporates queuine into the first position in the anticodon of four tRNA isoacceptors. Rat brain protein kinase C (PKC) was shown to stimulate rat liver TG Rase activity, TGRase preparations derived from rat liver have been observed to decrease in activity over time in storage at -20 or -70°C, Contamination of the samples by phosphatases was indicated by a p-nitrophenylphosphate conversion test, The addition of micromolar concentrations of the phosphatase inhibitors sodium pyrophosphate and sodium fluoride into TGRase isolation buffers resulted in a greater return of TGRase activity than without these inhibitors, Inactive TGRase preparations were reactivated to their original activity with the addition of PKC, In assays combining both TGRase and PKC enzymes, inhibitors of protein kinase C (sphingosine, staurosporine, H-7 and calphostin C) all blocked the reactivation of TGRase, whereas activators of protein kinase C (calcium, diacylglycerol and phosphatidyl serine) increased the activity of TGRase, None of the PKC modulators affected TGRase activity directly, Alkaline phosphatase, when added to assays, decreased the activity of TGRase and also blocked the reactivation of TGRase with PKC, Denaturing PAGE and autoradiography was performed on TGRase isolates that had been labelled with 32P by PKC, The resulting strong 60 kDa band (containing the major site for phosphorylation) and weak 34.5 kDa band (containing the TGRase activity) are suggested to associate to make up a 104 kDa heterodimer that comprises the TGRase enzyme, This was corroberated by native and denaturing size-exclusion chromatography These results suggest that PKC-dependent phosphorylation of TGRase is tied to efficient enzymatic function and therefore control of the queuine modification of tRNA

    Scenarios of habitat management options to reduce predator impacts on nesting waders

    Get PDF
    1. Wetland ecosystems throughout the world are threatened by drainage and intensification of agriculture. Consequently, many wetland species of conservation concern are now restricted to fewer and smaller sites, and maintaining these species often requires intensive habitat management.  2. In Western Europe, breeding wader populations have declined severely as a result of wetland degradation, but very high levels of predation on eggs and chicks are now preventing population recovery. Wet grassland management for breeding waders has focussed on providing suitable nesting habitats, but the potential for management of landscape features to influence predation rates is largely unknown.  3. Using a 7-year study of breeding lapwing, Vanellus vanellus, and redshank, Tringa totanus, we first identify features that influence nest predation, and then use this information to compare the magnitude of change in nest predation rates that could potentially result from future landscape management scenarios.  4. As lapwing nest predation rates are higher (a) in fields further from patches of tall vegetation, (b) close (<50 m) to field edges in wet fields, (c) further from field edges in dry fields and (d) in areas of low lapwing nesting density, we modelled a series of realistic scenarios in which the area of tall vegetation and the extent and distribution of surface water were varied across the reserve, in order to quantify the magnitude of change in nest predation rate that could potentially have been achieved through management.  5. Modelled scenarios of changes in surface water and area of tall vegetation indicated that reduced surface flooding combined with removal of tall vegetation could result in significant increases in lapwing nest predation rates in areas with low nesting densities and nests in field centres. By contrast a ~20% reduction in nest predation, corresponding to ~100 more chicks hatching per year, is predicted in scenarios with expansion of tall vegetation in areas with high lapwing nest density and nests close to field edges.  6. Synthesis and applications: These management scenarios suggest that, for breeding waders in wet grassland landscapes, creating areas of tall vegetation and concentrating surface flooding (to encourage high nesting densities and influence nesting distribution) can potentially help to reduce the unsustainably high levels of nest predation that are preventing population recovery

    Insights into toxic prymnesium parvum blooms:The role of sugars and algal viruses

    Get PDF
    Prymnesium parvum is a toxin-producing microalga that causes harmful algal blooms globally, which often result in large-scale fish kills that have severe ecological and economic implications. Although many toxins have previously been isolated from P. parvum, ambiguity still surrounds the responsible ichthyotoxins in P. parvum blooms and the biotic and abiotic factors that promote bloom toxicity. A major fish kill attributed to P. parvum occurred in Spring 2015 on the Norfolk Broads, a low-lying set of channels and lakes (Broads) found on the East of England. Here, we discuss how water samples taken during this bloom have led to diverse scientific advances ranging from toxin analysis to discovery of a new lytic virus of P. parvum, P. parvum DNA virus (PpDNAV-BW1). Taking recent literature into account, we propose key roles for sialic acids in this type of viral infection. Finally, we discuss recent practical detection and management strategies for controlling these devastating blooms

    Spin relaxation of conduction electrons in bulk III-V semiconductors

    Full text link
    Spin relaxation time of conduction electrons through the Elliot-Yafet, D'yakonov-Perel and Bir-Aronov-Pikus mechanisms is calculated theoretically for bulk GaAs, GaSb, InAs and InSb of both nn- and pp-type. Relative importance of each spin relaxation mechanism is compared and the diagrams showing the dominant mechanism are constructed as a function of temperature and impurity concentrations. Our approach is based upon theoretical calculation of the momentum relaxation rate and allows understanding of the interplay between various factors affecting the spin relaxation over a broad range of temperature and impurity concentration.Comment: an error in earlier version correcte

    Hand and Eye Dominance in Sport: Are Cricket Batters Taught to Bat Back-to-Front?

    Get PDF
    Background: When first learning to bimanually use a tool to hit a target (e.g., when chopping wood or hitting a golf ball), most people assume a stance that is dictated by their dominant hand. By convention, this means that a ‘right-handed’ or ‘left-handed’ stance that places the dominant hand closer to the striking end of the tool is adopted in many sports. Objective: The aim of this study was to investigate whether the conventional stance used for bimanual hitting provides the best chance of developing expertise in that task. Methods: Our study included 43 professional (international/first-class) and 93 inexperienced (<5 years’ experience) cricket batsmen. We determined their batting stance (plus hand and eye dominance) to compare the proportion of batters who adopted a reversed stance when batting (that is, the opposite stance to that expected based on their handedness). Results: We found that cricket batsmen who adopted a reversed stance had a stunning advantage, with professional batsmen 7.1 times more likely to adopt a reversed stance than inexperienced batsmen, independent of whether they batted right or left handed or the position of their dominant eye. Conclusion: Findings imply that batsmen who adopt a conventional stance may inadvertently be batting ‘back-to-front’ and have a significant disadvantage in the game. Moreover, the results may generalize more widely, bringing into question the way in which other bimanual sporting actions are taught and performed

    Assessing the toxicity and mitigating the impact of harmful Prymnesium blooms in eutrophic waters of the Norfolk Broads

    Get PDF
    Prymnesium parvum is a toxin-producing microalga, which causes harmful algal blooms globally, frequently leading to massive fish kills that have adverse ecological and economic implications for natural waterways and aquaculture alike. The dramatic effects observed on fish are thought to be due to algal polyether toxins, known as the prymnesins, but their lack of environmental detection has resulted in an uncertainty about the true ichthyotoxic agents. Using qPCR, we found elevated levels of P. parvum and its lytic virus, PpDNAV-BW1, in a fish-killing bloom on the Norfolk Broads, United Kingdom, in March 2015. We also detected, for the first time, the B-type prymnesin toxins in Broads waterway samples and gill tissue isolated from a dead fish taken from the study site. Furthermore, Norfolk Broads P. parvum isolates unambiguously produced B-type toxins in laboratory-grown cultures. A 2 year longitudinal study of the Broads study site showed P. parvum blooms to be correlated with increased temperature and that PpDNAV plays a significant role in P. parvum bloom demise. Finally, we used a field trial to show that treatment with low doses of hydrogen peroxide represents an effective strategy to mitigate blooms of P. parvum in enclosed water bodies
    • …
    corecore