Mean (± S.E.) prevalence of midgut infections in male <i>G. m. morsitans</i> (<i>Gmm</i>) or <i>G. p. palpalis</i> (<i>Gpp</i>) after RNAi knockdown using ds<i>tsetse EP</i>.

<p>Controls were either ds<i>Ampicillin</i>, ds<i>eGFP</i> or nuclease free water*. Flies were infected with either <i>T. b. brucei</i> TSW196 or <i>T. congolense</i> 1/148 (italics) blood stream forms in the indicated bloodmeal.</p

Immunoblot film lies below the nigrosine-stained PVDF.

<p>There is a decline in tsetse EP protein levels in midguts over a 7 day starvation period following the 5^th bloodmeal. 24 h  =  Flies starved for 7 days, fed a blood meal and then sacrificed 24 hours later. L  =  molecular mass ladder. Midgut proteins (1/2 midgut equivalent from pool of 5) were blotted with mAb 247.</p

Variant Surface Glycoproteins (VSGs) discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients confirmed to have late-stage human African trypanosomiasis.

<p>Variant Surface Glycoproteins (VSGs) discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients confirmed to have late-stage human African trypanosomiasis.</p

List of human plasma proteins with mass spectrometric molar intensities similar to those from the most abundant trypanosome proteins found in plasma from sleeping sickness patients.

*<p>Protein concentrations were retrieved from reference <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071463#pone.0071463-Hortin1" target="_blank">[19]</a>. Human proteins reported to be depleted by the LC20 IgY14 and Supermix LC10 columns <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0071463#pone.0071463-Patel1" target="_blank">[26]</a> have been omitted from this table.</p

Summary of patient information.

<p>Plasma and CSF were collected from <i>T. b. rhodesiense</i>-infected patients confirmed to have late stage human African trypanosomiasis at Lwala Hospital in central Uganda.</p

Chaperones and protein isomerases identified in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.

<p>Chaperones and protein isomerases identified in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.</p

Proteases and ubiquitin proteins discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.

<p>Proteases and ubiquitin proteins discovered in pooled plasma from <i>T. b. rhodesiense</i>-infected patients with parasitologically confirmed late stage human African trypanosomiasis.</p

ELISA detection of trypanosome antigens in plasma from <i>T. b. rhodesiense</i> -infected patients with late-stage human African trypanosomisis.

<p>All samples were tested in duplicate and the average response is shown. Error bars represent 1 standard deviation.</p

Analysis of trypanosomal mitochondrial activity by fluorescence microscopy and flow cytometry before and after treatment with BMAP-18.

<p><i>T. b. brucei</i> 427.01 PCF were incubated with rhodamine123 as described in the <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0000373#s2" target="_blank">Materials and Methods</a>, examined by fluorescence microscopy and digitally photographed (Panel A1). The rhodamine 123 labeled trypanosomes were then treated with a low dose of BMAP-18 (5 µg/mL) and photographed again (Panel A2). Rhodamine fluorescence (FL-2) was also monitored by flow cytometry, before and after varying intervals of BMAP-18 treatment (Panel B). The different colours indicate different incubation times with the BMAP-18 peptide. Red: diluent control, 30 min; Yellow: BMAP-18 treated, 5 min; Green: BMAP-18 treated, 10 min; Light blue: BMAP-18 treated, 20 min; Dark blue: BMAP-18 treated, 30 min. The plot clearly shows decreasing rhodamine 123 fluorescence as the incubation period of BMAP-18 with the trypanosomes was extended from 0–30 minutes. A dot-plot comparison of this experiment (Panel C) showed the effects of BMAP-18 treatment on fluorescence intensity vs side scatter (cell granularity) over time. The numbers in the upper right quadrants of the dot-plots indicate the percentage of healthy cells with strong mitochondrial rhodamine fluorescence. In Panel B, the horizontal (X) axis indicates arbitrary fluorescence units and the vertical (Y) axis indicates the number of cells (events). In Panel C, the Y axis is SSC (side scatter).</p

Multiple sequence alignment of the <i>T</i>. <i>congolense</i> calflagins.

<p>The positions of the two MS-identified tryptic peptides identified by mass spectrometry are highlighted in yellow and red boxes. The lower case, white highlighted v represent amino acid (valine-isoleucine) differences in two of the ORFs.</p