Role of copper efflux in pneumococcal pathogenesis and resistance to macrophage-mediated immune clearance

In bacteria, the intracellular levels of metals are mediated by tightly controlled acquisition and efflux systems. This is particularly true of copper, a trace element that is universally toxic in excess. During infection, the toxic properties of copper are exploited by the mammalian host to facilitate bacterial clearance. To better understand the role of copper during infection, we characterized the contribution of the cop operon to copper homeostasis and virulence in Streptococcus pneumoniae. Deletion of either the exporter, encoded by copA, or the chaperone, encoded by cupA, led to hypersensitivity to copper stress. We further demonstrated that loss of the copper exporter encoded by copA led to decreased virulence in pulmonary, intraperitoneal, and intravenous models of infection. Deletion of copA resulted in enhanced macrophage-mediated bacterial clearance in vitro. The attenuation phenotype of the copA mutant in the lung was found to be dependent on pulmonary macrophages, underscoring the importance of copper efflux in evading immune defenses. Overall, these data provide insight into the role of the cop operon in pneumococcal pathogenesis

Wheat and Maize Flour Fortification: Practical Recommendations for National Application

Joint statement by the World Health Organization, Food and Agriculture Organization of the United Nations, The United Nations Children s Fund, Global Alliance for Improved Nutrition, The Micronutrient Initiative and The Flour Fortification InitiativeREPRODUCTIVE HEALTHSURVEILLANCE AND INVESTIGATIONCURREN

New measures to chart toddlers' speech perception and language development : a test of the lexical restructuring hypothesis

Language acquisition factors at work in toddlers between 2 1/2 and 3 years of age were investigated in the first longitudinal study of this kind. New age-appropriate tasks were devised to measure the development of vocabulary size; articulation accuracy; sensitivity to the phonemic features of, in this case, Australian English; and the degree of specialisation towards the native tongue, as measured by language-specific speech perception; LSSP, with 45 Australian English learning toddlers (18 male, 27 female) at 30, 33, and 36 months of age. Results indicated (i) that nearly all measures improved linearly over age; (ii) that there were significant correlations between articulation ability and vocabulary size; and (iii) that, in confirmation of the lexical restructuring hypothesis, vocabulary size is significantly predicted by the broad range of native language abilities under the rubric of Phoneme Sensitivity, but not by the more specific measure of LSSP

OZI : Australian English communicative development inventory

For more than 20 years, the MacArthur–Bates Communicative Development Inventory (CDI) and its adaptations for languages other than English have been used as reliable measures of infants’ and toddlers’ early receptive and productive vocabulary size. This article introduces the OZI, the Australian English adaptation of the MacArthur–Bates CDI, now normed for 12- to 30-month-old children. The findings of two studies are presented: (1) a comparison study that demonstrated that toddlers (N = 64) acquiring Australian English (24- and 30-month-olds) obtain higher productive vocabulary scores on the OZI than the MacArthur–Bates CDI; and (2) an OZI norming study that included 12- to 30-month-old Australian infants and toddlers (N = 1496). These studies provide (i) evidence for the greater applicability of the OZI for infants and toddlers learning Australian English and (ii) productive vocabulary acquisition norms for Australian English for ages 12–30 months, a research and diagnostic tool highly anticipated by researchers and clinicians around Australia

Universality and language-specific experience in the perception of lexical tone and pitch

Two experiments focus on Thai tone perception by native speakers of tone languages (Thai, Cantonese, and Mandarin), a pitch–accent (Swedish), and a nontonal (English) language. In Experiment 1, there was better auditory-only and auditory–visual discrimination by tone and pitch–accent language speakers than by nontone language speakers. Conversely and counterintuitively, there was better visual-only discrimination by nontone language speakers than tone and pitch–accent language speakers. Nevertheless, visual augmentation of auditory tone perception in noise was evident for all five language groups. In Experiment 2, involving discrimination in three fundamental frequency equivalent auditory contexts, tone and pitch–accent language participants showed equivalent discrimination for normal Thai speech, filtered speech, and violin sounds. In contrast, nontone language listeners had significantly better discrimination for violin sounds than filtered speech and in turn speech. Together the results show that tone perception is determined by both auditory and visual information, by acoustic and linguistic contexts, and by universal and experiential factors

Dynamic Pneumococcal Genetic Adaptations Support Bacterial Growth and Inflammation during Coinfection with Influenza

Streptococcus pneumoniae (pneumococcus) is one of the primary bacterial pathogens that complicates influenza virus infections. These bacterial coinfections increase influenza-associated morbidity and mortality through a number of immunological and viral-mediated mechanisms, but the specific bacterial genes that contribute to postinfluenza pathogenicity are not known. Here, we used genome-wide transposon mutagenesis (Tn-Seq) to reveal bacterial genes that confer improved fitness in influenza virus-infected hosts. The majority of the 32 genes identified are involved in bacterial metabolism, including nucleotide biosynthesis, amino acid biosynthesis, protein translation, and membrane transport. We generated mutants with single-gene deletions (SGD) of five of the genes identified, SPD1414, SPD2047 (cbiO1), SPD0058 (purD), SPD1098, and SPD0822 (proB), to investigate their effects on in vivo fitness, disease severity, and host immune responses. The growth of the SGD mutants was slightly attenuated in vitro and in vivo, but each still grew to high titers in the lungs of mock- and influenza virus-infected hosts. Despite high bacterial loads, mortality was significantly reduced or delayed with all SGD mutants. Time-dependent reductions in pulmonary neutrophils, inflammatory macrophages, and select proinflammatory cytokines and chemokines were also observed. Immunohistochemical staining further revealed altered neutrophil distribution with reduced degeneration in the lungs of influenza virus-SGD mutant-coinfected animals. These studies demonstrate a critical role for specific bacterial genes and for bacterial metabolism in driving virulence and modulating immune function during influenza-associated bacterial pneumonia.Open access articleThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Nuclear DBF-2-related Kinases Are Essential Regulators of Cytokinesis in Bloodstream Stage Trypanosoma brucei

Nuclear DBF-2-related (NDR) kinases are essential regulators of cell cycle progression, growth, and development in many organisms and are activated by the binding of an Mps One Binder (MOB) protein partner, autophosphorylation, and phosphorylation by an upstream STE20 family kinase. In the protozoan parasite, Trypanosoma brucei, the causative agent of human African trypanosomiasis, the NDR kinase, PK50, is expressed in proliferative life cycle stages and was shown to complement a yeast NDR kinase mutant cell line. However, the function of PK50 and a second NDR kinase, PK53, in T. brucei has not been determined to date, although trypanosome MOB1 is known to be essential for cytokinesis, suggesting the NDR kinases may also be involved in this process. Here, we show that specific depletion of PK50 or PK53 from bloodstream stage trypanosomes resulted in the rapid accumulation of cells with two nuclei and two kinetoplasts, indicating that cytokinesis was specifically inhibited. This led to a deregulation of the cell cycle and cell death and provides genetic validation of these kinases as potential novel drug targets for human African trypanosomiasis. Recombinant active PK50 and PK53 were produced and biochemically characterized. Both enzymes autophosphorylated, were able to trans-phosphorylate generic kinase substrates in vitro, and were active in the absence of phosphorylation by an upstream kinase. Additionally, both enzymes were active in the absence of MOB1 binding, which was also demonstrated to likely be a feature of the kinases in vivo. Biochemical characterization of recombinant PK50 and PK53 has revealed key kinetic differences between them, and the identification of in vitro peptide substrates in this study paves the way for high throughput inhibitor screening of these kinases