11 research outputs found
Prenyl pyrophosphate stimulation increased cellular tolerance to H<sub>2</sub>O<sub>2</sub>-induced oxidative stress.
<p><i>A. gambiae</i> 4a3B cells were seeded in 96-well plates. (<b>A</b>) HMBPP or IPP (10 µM) were added at indicated time points prior to oxidative insult (n = 3). (<b>B</b>) Cells were preincubated for 3 h with HMBPP at different concentrations (n = 6-8). At 30 min before H<sub>2</sub>O<sub>2</sub> addition, cells were loaded with (<b>A</b>) CellROX Deep Red Reagent or (<b>B</b>) DCFDA for oxidative stress detection. Cells were washed and challenged with H<sub>2</sub>O<sub>2</sub> for 2 h. Bars represent mean relative fluorescence ± SE, asterisks denote significant differences (*P < 0.05, **P < 0.01, *** P < 0.001) compared to H<sub>2</sub>O<sub>2</sub> control.</p
Temporal expression of selected immune genes relative to control blood meal as determined by qRT-PCR.
<p>(<b>A</b>–<b>E</b>) Bars represent mean ± SE from at least three (HMBPP) or two (PGN) independent experiments. Whole bodies were used from 5–10 fully engorged females. Asterisks (*<i>P</i> < 0.05, **<i>P</i> < 0.01) denote significantly different expression compared to control feed.</p
Expression in midgut and carcass and abundance of midgut bacteria relative to control feed.
<p>(<b>A</b>–<b>B</b>) At 3 and 6 hpi, 5-10 midguts and corresponding carcasses per sample were dissected and subjected to qRT-PCR analysis of DEF1, DUOX and NOS. (<b>C</b>–<b>D</b>) Effects of HMBPP on relative midgut bacterial 16S were quantified using qPCR. (<b>C</b>) At 24 hpi, bacterial gDNA was extracted from midguts (n = 4). (<b>D</b>) Midgut cDNA from 3 and 6 hpi (n = 3). Bars represent mean ± SE (*<i>P</i> < 0.05, ** <i>P</i> < 0.01).</p
HMBPP activates phosphorylation of JNK, p38 and FOXO in <i>A. gambiae</i> 4a3B cells.
<p>(<b>A</b>–<b>D</b>) Mean fold changes ± SE relative to control stimulation and representative western blot images of target phosphoproteins. Membranes where stripped and reprobed for GAPDH to normalize relative band intensities. Asterisks represent significant differences (*<i>P</i> < 0.05, *** <i>P</i> < 0.001) from at least three independent experiments.</p
Insights from the Genome Annotation of <i>Elizabethkingia anophelis</i> from the Malaria Vector <i>Anopheles gambiae</i>
<div><p><i>Elizabethkingia anophelis</i> is a dominant bacterial species in the gut ecosystem of the malaria vector mosquito <i>Anopheles gambiae</i>. We recently sequenced the genomes of two strains of <i>E. anophelis</i>, R26<sup>T</sup> and Ag1, isolated from different strains of <i>A. gambiae</i>. The two bacterial strains are identical with a few exceptions. Phylogenetically, <i>Elizabethkingia</i> is closer to <i>Chryseobacterium</i> and <i>Riemerella</i> than to <i>Flavobacterium.</i> In line with other Bacteroidetes known to utilize various polymers in their ecological niches, the <i>E. anophelis</i> genome contains numerous TonB dependent transporters with various substrate specificities. In addition, several genes belonging to the polysaccharide utilization system and the glycoside hydrolase family were identified that could potentially be of benefit for the mosquito carbohydrate metabolism. In agreement with previous reports of broad antibiotic resistance in <i>E. anophelis</i>, a large number of genes encoding efflux pumps and β-lactamases are present in the genome. The component genes of resistance-nodulation-division type efflux pumps were found to be syntenic and conserved in different taxa of Bacteroidetes. The bacterium also displays hemolytic activity and encodes several hemolysins that may participate in the digestion of erythrocytes in the mosquito gut. At the same time, the OxyR regulon and antioxidant genes could provide defense against the oxidative stress that is associated with blood digestion. The genome annotation and comparative genomic analysis revealed functional characteristics associated with the symbiotic relationship with the mosquito host.</p></div
Isoprenoid synthesis pathway in four bacterial species.
<p>The color code represents the enzymes that are present in the species.</p
The similarity of comparable CDS between R26<sup>T</sup> and related genomes in the family Flavobacteriaceae.
<p>Numbers within parenthesis reflect the percentage of total comparable CDS.</p
Graphic view of a syntenic gene cluster that is conserved in five taxa of Bacteroidetes.
<p>The genes encoding the components of TonB dependent transporters are ExbB, ExbD and TonB. Locus ID of each gene is given in the boxes, with taxon prefix (e.g. D505_ for <i>E.a.</i>) in the ExbB box for each species. The box with * in <i>D. fermentans</i> represents a predicted gene encoding a hypothetic protein. The scale bar represents 1 kb in length. Phylogenetically, <i>E. anophelis</i> and <i>F. johnsoniae</i> belong to the class Flavobacteria, <i>Dyadobacter fermentans</i> belongs to the class Cytophagia, <i>Arcticibacter svalbardensis</i> is in the class Sphingobacteriia, and <i>Bacteroides thetaiotaomicron</i> is located in the class Bacteroidia.</p
Subsystem category distribution statistics for the genome of <i>E. Anophelis</i> as annotated by RAST.
<p>The pie chart represents relative abundance of each subsystem category and numbers depict subsystem feature counts.</p
Summary of the functional subcategories of resistance genes.
<p>Average identity reflects the mean identity of genes within the subcategory compared to R26<sup>T</sup>. <i>E. m</i>., <i>Elizabethkingia meningoseptica</i>; <i>C. g.</i>, <i>Chryseobacterium gleum</i>; <i>F. b., Flavobacterium branchiophilum</i>.</p