Structural basis of metallo-β-lactamase, serine-β-lactamase and penicillin-binding protein inhibition by cyclic boronates

β-Lactamases enable resistance to almost all β-lactam antibiotics. Pioneering work revealed that acyclic boronic acids can act as ‘transition state analogue’ inhibitors of nucleophilic serine enzymes, including serine-β-lactamases. Here we report biochemical and biophysical analyses revealing that cyclic boronates potently inhibit both nucleophilic serine and zinc-dependent β-lactamases by a mechanism involving mimicking of the common tetrahedral intermediate. Cyclic boronates also potently inhibit the non-essential penicillin-binding protein PBP 5 by the same mechanism of action. The results open the way for development of dual action inhibitors effective against both serine- and metallo-β-lactamases, and which could also have antimicrobial activity through inhibition of PBPs

Mechanisms Underlying Interferon-γ-Induced Priming of Microglial Reactive Oxygen Species Production.

Microglial priming and enhanced reactivity to secondary insults cause substantial neuronal damage and are hallmarks of brain aging, traumatic brain injury and neurodegenerative diseases. It is, thus, of particular interest to identify mechanisms involved in microglial priming. Here, we demonstrate that priming of microglia with interferon-γ (IFN γ) substantially enhanced production of reactive oxygen species (ROS) following stimulation of microglia with ATP. Priming of microglial ROS production was substantially reduced by inhibition of p38 MAPK activity with SB203580, by increases in intracellular glutathione levels with N-Acetyl-L-cysteine, by blockade of NADPH oxidase subunit NOX2 activity with gp91ds-tat or by inhibition of nitric oxide production with L-NAME. Together, our data indicate that priming of microglial ROS production involves reduction of intracellular glutathione levels, upregulation of NADPH oxidase subunit NOX2 and increases in nitric oxide production, and suggest that these simultaneously occurring processes result in enhanced production of neurotoxic peroxynitrite. Furthermore, IFNγ-induced priming of microglial ROS production was reduced upon blockade of Kir2.1 inward rectifier K+ channels with ML133. Inhibitory effects of ML133 on microglial priming were mediated via regulation of intracellular glutathione levels and nitric oxide production. These data suggest that microglial Kir2.1 channels may represent novel therapeutic targets to inhibit excessive ROS production by primed microglia in brain pathology

Modes of Aβ toxicity in Alzheimer’s disease

Alzheimer’s disease (AD) is reaching epidemic proportions, yet a cure is not yet available. While the genetic causes of the rare familial inherited forms of AD are understood, the causes of the sporadic forms of the disease are not. Histopathologically, these two forms of AD are indistinguishable: they are characterized by amyloid-β (Aβ) peptide-containing amyloid plaques and tau-containing neurofibrillary tangles. In this review we compare AD to frontotemporal dementia (FTD), a subset of which is characterized by tau deposition in the absence of overt plaques. A host of transgenic animal AD models have been established through the expression of human proteins with pathogenic mutations previously identified in familial AD and FTD. Determining how these mutant proteins cause disease in vivo should contribute to an understanding of the causes of the more frequent sporadic forms. We discuss the insight transgenic animal models have provided into Aβ and tau toxicity, also with regards to mitochondrial function and the crucial role tau plays in mediating Aβ toxicity. We also discuss the role of miRNAs in mediating the toxic effects of the Aβ peptide

Evaluation of Amendments to Control Phosphorus Losses in Runoff from Dairy-Soiled Water

peer-reviewedAmendments with the potential to reduce phosphorus (P) losses from agricultural grassland arising from the land application of dairy-soiled water (DSW) were investigated. Optimal application rates were studied, and associated costs and feasibility were estimated. First, batch tests were carried out to identify appropriate chemicals or phosphorus sorbing materials to control P in runoff from DSW. Then, the best four treatments were examined in an agitator test. In this test, soil—placed in a beaker—was loaded with DSW or amended DSW at a rate equivalent to 5 mm ha−1 (the maximum permissible application rate of DSW allowable in a 42-day period in Ireland). The soil was overlain with continuously stirred water to simulate runoff on land-applied DSW. Optimum application rates were selected based on percentage removal of dissolved reactive phosphorus in overlying water and the estimated cost of amendment. The costs of the amendments, per cubic metre of DSW, increased in the order: bottom ash (1.55 €), alum (1.67 to 1.92 €), FeCl2·4H2O (3.55 to 8.15 €), and lime (20.31 to 88.65 €). The feasibility of the amendments, taking into account their cost, potential adverse effects, public perception, and their performance, decreased in the order: alum > FeCl2·4H2O > bottom ash > lime. Amendments to DSW could be introduced in critical source areas—areas where high soil test P and direct migration pathways to a receptor overlap.Department of Agriculture, Fisheries and Food (Ireland) Research Stimulus Programme 2007 (RSF 07 525