Shannon-Weaver indices compared among samples collected in dry and rainy season.

<p>Crosses in the box indicate the mean, and black lines are medians.</p

Distribution of mosquitoes dissected over two collection periods; dry and rainy seasons.

<p>Numbers in brackets are the number of midgut pools.</p

The progressive stages of development of BU in mouse injected in the right footpad with <i>M</i>. <i>ulcerans</i>-infected <i>A</i>. <i>polyphaga</i>.

<p>Panel A shows footpad 1 dpi. Panel B shows footpad with erythema 3 dpi. Panel C shows footpad with edema 7 dpi. Panel D shows swollen footpad and thigh 25 dpi. The photographs are typically representatives of each group (n = 3).</p

Rapid Turnover of Long Noncoding RNAs and the Evolution of Gene Expression

<div><p>A large proportion of functional sequence within mammalian genomes falls outside protein-coding exons and can be transcribed into long RNAs. However, the roles in mammalian biology of long noncoding RNA (lncRNA) are not well understood. Few lncRNAs have experimentally determined roles, with some of these being lineage-specific. Determining the extent by which transcription of lncRNA loci is retained or lost across multiple evolutionary lineages is essential if we are to understand their contribution to mammalian biology and to lineage-specific traits. Here, we experimentally investigated the conservation of lncRNA expression among closely related rodent species, allowing the evolution of DNA sequence to be uncoupled from evolution of transcript expression. We generated total RNA (RNAseq) and H3K4me3-bound (ChIPseq) DNA data, and combined both to construct catalogues of transcripts expressed in the adult liver of <em>Mus musculus domesticus</em> (C57BL/6J), <em>Mus musculus castaneus</em>, and <em>Rattus norvegicus</em>. We estimated the rate of transcriptional turnover of lncRNAs and investigated the effects of their lineage-specific birth or death. LncRNA transcription showed considerably greater gain and loss during rodent evolution, compared with protein-coding genes. Nucleotide substitution rates were found to mirror the <em>in vivo</em> transcriptional conservation of intergenic lncRNAs between rodents: only the sequences of noncoding loci with conserved transcription were constrained. Finally, we found that lineage-specific intergenic lncRNAs appear to be associated with modestly elevated expression of genomically neighbouring protein-coding genes. Our findings show that nearly half of intergenic lncRNA loci have been gained or lost since the last common ancestor of mouse and rat, and they predict that such rapid transcriptional turnover contributes to the evolution of tissue- and lineage-specific gene expression.</p> </div

Experimental demonstration of the possible role of <i>Acanthamoeba polyphaga</i> in the infection and disease progression in Buruli Ulcer (BU) using ICR mice

<div><p>The transmission of Buruli ulcer (BU), caused by <i>Mycobacterium ulcerans</i> (<i>MU</i>), remains puzzling although a number of hypothesis including through bites of infected aquatic insects have been proposed. We report the results of experiments using ICR mice that give credence to our hypothesis that <i>Acanthamoeba</i> species may play a role in BU transmission. We cocultured <i>MU</i> N2 and <i>MU</i> 1615 which expresses red fluorescent protein (RFP) and <i>Acanthamoeba polyphaga</i> (<i>AP</i>), and confirmed infected <i>AP</i> by Ziehl-Neelsen (ZN) staining. We tested for viability of <i>MU</i> inside <i>AP</i> and observed strong RFP signals inside both trophozoites and cysts after 3 and 42 days of coculturing respectively. ICR mice were topically treated, either on shaved intact or shaved pinpricked rumps, with one of the following; <i>MU</i> N2 only (2.25 x 10⁶ colony forming units [CFU] / ml), <i>MU</i> N2:<i>AP</i> coculture (2.96 x 10⁴ CFU: 1.6 x 10⁶ cells/ml), <i>AP</i> only (1.6 x 10⁶ cells/ml), PYG medium and sterile distilled water. Both <i>MU</i> N2 only and <i>MU</i> N2:<i>AP</i> elicited reddening on day (D) 31; edema on D 45 and D 44 respectively, and ulcers on D 49 at pinpricked sites only. To ascertain infectivity and pathogenicity of <i>MU</i> N2 only and <i>MU</i> N2:<i>AP</i>, and compare their virulence, the standard mouse footpad inoculation method was used. <i>MU</i> N2:<i>AP</i> elicited reddening in footpads by D 3 compared to D 14 with <i>MU</i> N2 only of the same dose of <i>MU</i> N2 (2.96 x 10⁴ CFU). ZN-stained <i>MU</i> were observed in both thin sectioned and homogenized lesions, and aspirates from infected sites. Viable <i>MU</i> N2 were recovered from cultures of the homogenates and aspirates. This study demonstrates in ICR mice <i>MU</i> transmission via passive infection, and shows that punctures in the skin are prerequisite for infection, and that coculturing of <i>MU</i> with <i>AP</i> enhances pathogenesis.</p></div

Distribution of Ala147Thr genotypes by sex among cases and non-cases in studies included in the meta-analysis.

<p>Distribution of Ala147Thr genotypes by sex among cases and non-cases in studies included in the meta-analysis.</p

Visualization of <i>M</i>. <i>ulcerans</i> within <i>A</i>. <i>polyphaga</i> trophozoites.

<p>Panel A shows ZN stain of <i>A</i>. <i>polyphaga</i> trophozoites. No acid-fast particle was seen in the axenic culture of <i>A</i>. <i>polyphaga</i> trophozoites. Panel B shows a ZN stain of <i>A</i>. <i>polyphaga</i> trophozoites infected with <i>M</i>. <i>ulcerans</i>. <i>A</i>. <i>polyphaga</i> trophozoites stained bluish and <i>M</i>. <i>ulcerans</i> stained reddish- pink (acid-fast particles). Acid fast particles denoting the presence of <i>M</i>. <i>ulcerans</i> can be seen inside trophozoites of <i>A</i>. <i>polyphaga</i> infected with <i>M</i>. <i>ulcerans</i>. The average rate of infection was 47% ± 8.6 (SD) (average counts in 5 fields randomly selected under 200x magnification).</p

Sex-specific meta-analysis of the <i>CPB2</i> Ala147Thr variant using the dominant model and risk of venous thrombosis.

<p>The analysis was stratified by sex. The solid squares represent the ORs from the individual studies; horizontal lines represent corresponding CIs; the diamonds show the combined ORs.</p

The progression of BU signs in the various treatment groups post-inoculation.

<p>The progression of BU signs in the various treatment groups post-inoculation.</p

The progressive development of BU in mouse topically treated with <i>M</i>. <i>ulcerans</i>-infected <i>A</i>. <i>polyphaga</i> at the punctured skin of the rump (lower back).

<p>Panel A shows site of inoculation 1 day post-inoculation (dpi). Panel B shows inflammation (erythema) at site of inoculation 31 dpi. Panel C shows an edema at site of inoculation 44 dpi. Panel D shows ulcer at the site of inoculation 49 dpi. Photograph is representative of group (n = 3).</p