Exopolysaccharides contribute significantly to attachment and biofilm formation in the opportunisitc pathogen Pseudomonas aeruginosa. The Psl polysaccharide, which is synthesized by the polysaccharide synthesis locus (psl), is required for biofilm formation in nonmucoid strains that do not rely on alginate as the principal biofilm polysaccharide. In-frame deletion and complementation studies of individual psl genes revealed that eleven psl genes, pslACDEFGHIJKL, are required for Psl production and surface attachment. We also present the first structural analysis of the psl-dependent polysaccharide, which consists of a repeating pentasaccharide containing d-mannose, d-glucose, and l-rhamnose: false[→3false)−normalβ−normalD−Manp2↑1α−D−Manp−false(1→3false)−normalβ−normalD−Manp−false(1→3false)−normalα−normalL−Rhap−false(1→3false)−normalβ−normalD−Glcp−false(1→false]normaln− In addition we identified the sugar nucleotide precursors involved in Psl generation and demonstrated the requirement for GDP-d-mannose, UDP-d-glucose, and dTDP-l-rhamnose in Psl production and surface attachment. Finally, genetic analyses revealed that wbpW restored Psl production in a pslB mutant and pslB promoted A-band LPS synthesis in a wbpW mutant, indicating functional redundancy and overlapping roles for these two enzymes. The structural and genetic data presented here provide a basis for further investigation of the Psl proteins and potential roles for Psl in the biology and pathogenesis of P. aeruginosa.

Mineral inclusions encapsulated in diamonds are the oldest, deepest, and most pristine samples of Earth's mantle. They provide age and chemical information over a period of 3.5 billion years--a span that includes continental crustal growth, atmospheric evolution, and the initiation of plate tectonics. We compiled isotopic and bulk chemical data of silicate and sulfide inclusions and found that a compositional change occurred 3.0 billion years ago (Ga). Before 3.2 Ga, only diamonds with peridotitic compositions formed, whereas after 3.0 Ga, eclogitic diamonds became prevalent. We suggest that this resulted from the capture of eclogite and diamond-forming fluids in subcontinental mantle via subduction and continental collision, marking the onset of the Wilson cycle of plate tectonics.

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The redox state of Earth's convecting mantle, masked by the lithospheric plates and basaltic magmatism of plate tectonics, is a key unknown in the evolutionary history of our planet. Here we report that large, exceptional gem diamonds like the Cullinan, Constellation, and Koh-i-Noor carry direct evidence of crystallization from a redox-sensitive metallic liquid phase in the deep mantle. These sublithospheric diamonds contain inclusions of solidified iron-nickel-carbon-sulfur melt, accompanied by a thin fluid layer of methane ± hydrogen, and sometimes majoritic garnet or former calcium silicate perovskite. The metal-dominated mineral assemblages and reduced volatiles in large gem diamonds indicate formation under metal-saturated conditions. We verify previous predictions that Earth has highly reducing deep mantle regions capable of precipitating a metallic iron phase that contains dissolved carbon and hydrogen.

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Otitis media is an extremely common pediatric ailment caused by opportunists that reside within the nasopharynx. Inflammation within the upper airway can promote ascension of these opportunists into the middle ear chamber. Otitis media can be chronic/recurrent in nature, and a wealth of data indicates that in these cases the bacteria persist within biofilms. Epidemiological data demonstrates most cases of otitis media are polymicrobial, which may have significant impact on antibiotic resistance. In this study, we used in vitro biofilm assays and rodent infection models to examine the impact of polymicrobial infection with Moraxella catarrhalis and Streptococcus pneumoniae (pneumococcus) on biofilm resistance to antibiotic treatment and persistence in vivo. Consistent with prior work, M. catarrhalis conferred beta-lactamase dependent passive protection from beta-lactam killing to pneumococci within polymicrobial biofilms. Moreover, pneumococci increased resistance of M. catarrhalis to macrolide killing in polymicrobial biofilms. However, pneumococci increased colonization in vivo by M. catarrhalis in a quorum signal-dependent manner. We also found that co-infection with M. catarrhalis affects middle ear ascension of pneumococci in both mice and chinchillas. Therefore, we conclude that residence of M. catarrhalis and pneumococci within the same biofilm community significantly impacts resistance to antibiotic treatment and bacterial persistence in vivo.