ICAR: endoscopic skull‐base surgery

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Taking care to correctly diagnose viral anterior uveitis

Lucy C. Crawford, Justine R. Smit

Natural Genetic Transformation Generates a Population of Merodiploids in Streptococcus pneumoniae.

Contains fulltext : 119152.pdf (publisher's version ) (Open Access)Partial duplication of genetic material is prevalent in eukaryotes and provides potential for evolution of new traits. Prokaryotes, which are generally haploid in nature, can evolve new genes by partial chromosome duplication, known as merodiploidy. Little is known about merodiploid formation during genetic exchange processes, although merodiploids have been serendipitously observed in early studies of bacterial transformation. Natural bacterial transformation involves internalization of exogenous donor DNA and its subsequent integration into the recipient genome by homology. It contributes to the remarkable plasticity of the human pathogen Streptococcus pneumoniae through intra and interspecies genetic exchange. We report that lethal cassette transformation produced merodiploids possessing both intact and cassette-inactivated copies of the essential target gene, bordered by repeats (R) corresponding to incomplete copies of IS861. We show that merodiploidy is transiently stimulated by transformation, and only requires uptake of a approximately 3-kb DNA fragment partly repeated in the chromosome. We propose and validate a model for merodiploid formation, providing evidence that tandem-duplication (TD) formation involves unequal crossing-over resulting from alternative pairing and interchromatid integration of R. This unequal crossing-over produces a chromosome dimer, resolution of which generates a chromosome with the TD and an abortive chromosome lacking the duplicated region. We document occurrence of TDs ranging from approximately 100 to approximately 900 kb in size at various chromosomal locations, including by self-transformation (transformation with recipient chromosomal DNA). We show that self-transformation produces a population containing many different merodiploid cells. Merodiploidy provides opportunities for evolution of new genetic traits via alteration of duplicated genes, unrestricted by functional selective pressure. Transient stimulation of a varied population of merodiploids by transformation, which can be triggered by stresses such as antibiotic treatment in S. pneumoniae, reinforces the plasticity potential of this bacterium and transformable species generally

A Streptococcus pneumoniae infection model in larvae of the wax moth Galleria mellonella

The bacterium Streptococcus pneumoniae is a leading human opportunistic pathogen. The limitations of the current vaccine have led to increased recognition of the need to understand bacterial behaviour and competitive dynamics using in vivo models of infection. Here, we investigate the potential application of the larvae of the wax moth Galleria mellonella as an informative infection model. Larvae were challenged with a range of doses of S. pneumoniae isolates differing in known virulence factors to determine the LD(50) values. Infection dynamics were determined by obtaining bacterial counts from larvae over a time course. Differences in virulence between serotypes could be distinguished in this host. Infection with strains differing in known virulence factors demonstrated predicted differences in virulence. Acapsulate and pneumolysin-negative strains were less virulent than their respective wild types. A large reduction in virulence was seen in strains lacking cell wall D-alanylation. The mortality of G. mellonella larvae is attributable to bacterial growth within larvae, while surviving larvae are able to clear infections by reducing bacterial numbers. These data demonstrate that G. mellonella larvae represent an in vivo infection model with applications for investigating aspects of bacterial-host interactions such as the role of antimicrobial peptide activity and resistance