Surface antigens and potential virulence factors from parasites detected by comparative genomics of perfect amino acid repeats-2

<p><b>Copyright information:</b></p><p>Taken from "Surface antigens and potential virulence factors from parasites detected by comparative genomics of perfect amino acid repeats"</p><p>http://www.proteomesci.com/content/5/1/20</p><p>Proteome Science 2007;5():20-20.</p><p>Published online 20 Dec 2007</p><p>PMCID:PMC2254594.</p><p></p> the remainders of the respective proteomes. Bars indicate the median. The organism with 30% of asparagines in the repeats in N-glycosylation consensus is

Surface antigens and potential virulence factors from parasites detected by comparative genomics of perfect amino acid repeats-0

<p><b>Copyright information:</b></p><p>Taken from "Surface antigens and potential virulence factors from parasites detected by comparative genomics of perfect amino acid repeats"</p><p>http://www.proteomesci.com/content/5/1/20</p><p>Proteome Science 2007;5():20-20.</p><p>Published online 20 Dec 2007</p><p>PMCID:PMC2254594.</p><p></p>s. Ag, ; At, ; Br, ; Ce, ; Dd, ; Dm, ; Hs, ; Kl, ; Mm, ; Rn, ; Sc, ; Sp, ; Yl, ; Ch, ; Cn, ; Ec, ; Eh, ; Gd, ; Lm, ; Pf, ; Ta, ; Tb, ; Tp, ; r, Spearman coefficient

Respiratory inhibitor-sensitivity in <i>T</i>. <i>b</i>. <i>gambiense</i> isolates and AQP-mediated glycerol transport.

<p>(A) SHAM EC₅₀ values for the <i>T</i>. <i>b gambiense</i> strains are indicated +/- glycerol. The inset shows pentamidine EC₅₀ values. * indicates significantly different (<i>P</i><0.05) to STIB930 using an ANOVA test in GraphPad Prism. All pairwise comparisons +/- 10 mM glycerol also indicated significant (<i>P</i> <0.001) differences using a Student’s <i>t</i>-test. Error bars, SD. (B) Propyl gallate and (C) Ascofuranone EC₅₀ values. Other details as in A. (D) Model for glycerol transport by AQPs in <i>T</i>. <i>b</i>. <i>gambiense</i>. The weight of the arrows indicates relative impact on glycerol utilisation and efflux, with AQP2 being the major contributor; note that transport across both the plasma and glycosomal membranes contributes to glycerol utilisation and efflux, see the text for more details. The right-hand panel indicates the situation in melarsoprol-resistant (reduced melarsoprol uptake) and SHAM-sensitive (reduced glycerol efflux) clinical isolates where a chimeric AQP2/3 replaces AQP2 and AQP3.</p

<i>aqp</i>-null <i>T</i>. <i>b</i>. <i>brucei</i> display defective glycerol-efflux and respiratory inhibitor-sensitivity.

<p>(A) Bloodstream <i>T</i>. <i>brucei</i> express a SHAM-sensitive mitochondrial trypanosome alternative oxidase (TAO). Under aerobic conditions, TAO activity allows ATP production without glycerol production as indicated by the black lines (left-hand blue ‘cell’). SHAM blocks TAO-activity, leading to the anaerobic production of glycerol, which is toxic if not removed, as indicated by the black lines (right-hand blue ‘cell’). SHAM dose-response curves for wild-type (WT) and <i>aqp1-2-3</i> null-cells. EC₅₀ values are indicated. (B) In the presence of SHAM and glycerol, the glycerol inhibits glycerol kinase (GK), also preventing ATP-production by the anaerobic route (blue ‘cell’). SHAM dose-response curves as in A but in the presence of 10 mM glycerol. (C) Propyl gallate and octyl gallate dose-response curves for wild-type (WT) and <i>aqp1-2-3</i> null-cells. EC₅₀ values are indicated. (D) SHAM EC₅₀ values +/- 10 mM glycerol from A-B and also from <i>aqp2</i>, <i>aqp2-3</i> and <i>aqp1-2-3</i> cells re-expressing ^GFPAQP2. * indicates significantly different (<i>P</i><0.01) to WT using an ANOVA test in GraphPad Prism. Pairwise comparisons +/- glycerol, except in the case of the <i>aqp1-2-3</i> null, indicated significant (<i>P</i> <0.001) differences using a Student’s <i>t</i>-test. Error bars, SD. The images to the right show re-expression of ^GFPAQP2 in <i>aqp1-2-3</i> null-cells.</p

Surface antigens and potential virulence factors from parasites detected by comparative genomics of perfect amino acid repeats-3

<p><b>Copyright information:</b></p><p>Taken from "Surface antigens and potential virulence factors from parasites detected by comparative genomics of perfect amino acid repeats"</p><p>http://www.proteomesci.com/content/5/1/20</p><p>Proteome Science 2007;5():20-20.</p><p>Published online 20 Dec 2007</p><p>PMCID:PMC2254594.</p><p></p> data are stored in a database that is accessible on-line [44] via the depicted interface (bottom). This allows user-defined Boolean queries for repeat-containing surface proteins

Glycerol uptake and utilisation is perturbed in <i>aqp</i>-null <i>T</i>. <i>b</i>. <i>brucei</i>.

<p>(A) ATP levels were assessed in the strains indicated after incubation in 5 mM glucose or glycerol. Readings were taken in triplicate and normalised to substrate only. * indicates significantly different (<i>P</i><0.001) to wild-type (WT) using an ANOVA test in GraphPad Prism. Error bars, SD. (B) Radiolabelled glycerol uptake was assessed in the strains indicated. Readings were taken in quadruplicate. * indicates significant difference (<i>P</i><0.05) using a Student’s <i>t</i>-test. Error bars, SD.</p

<i>T</i>. <i>b</i>. <i>brucei</i> tolerates the loss of all three <i>AQPs</i>.

<p>(A) The schematic maps indicate the <i>AQP1</i> and <i>AQP2-3</i> regions replaced by selectable markers as also indicated on the right. Δ indicates the regions deleted while the probes used for Southern blotting are shown above the maps. H, <i>Hpa</i>I; S, <i>Sac</i>II. (B) The Southern blots indicate deletion of the <i>AQP1</i> alleles in <i>aqp1</i> and three independent <i>aqp1-2-3</i> strains. Wild-type (WT) is shown for comparison. Genomic DNA was digested with <i>Hpa</i>I. (C) The Southern blots indicate deletion of the <i>AQP2-3</i> alleles in <i>aqp1-2-3</i> strains. WT is shown for comparison. Genomic DNA was digested with <i>Sac</i>II.</p

Final Report of the Evaluation of the Teaching and Learning Technology Programme

<p>^<i>a</i><i>L</i>. <i>don</i>. axen.: axenic amastigotes of <i>L</i>. <i>donovani</i>, strain MHOM-ET-67/L82.</p><p>^<i>b</i><i>L</i>. <i>don</i>. intracell: intracellular amastigotes of <i>L</i>. <i>donovani</i> strain MHOM-ET-67/L82.</p><p>^cCytotoxicity on macrophages infected with <i>L</i>. <i>donovani</i>.</p><p>^dCytotoxicity on peritoneal mouse macrophages.</p><p>^eSelectivity index: IC₅₀ Cytotoxicity macrophages/ IC₅₀<i>L</i>. <i>donovani</i>. IC₅₀ values are means of two independent assays, which varied < ±50%.</p><p><i>In vitro</i> activity against <i>L</i>. <i>donovani</i> in IC₅₀ (μM) of compounds fulfilling hit criteria.</p

<i>In vitro</i> activity against <i>T</i>. <i>cruzi</i> in IC₅₀ (μM) of compounds fulfilling hit criteria.

<p>^a<i>T</i>. <i>cruzi</i>, strain Tulahuen C4, intracellular amastigotes.</p><p>^bCytotoxicity on L6 cells.</p><p>^cSelectivity index: IC₅₀ Cytotoxicity L6/ IC₅₀<i>T</i>. <i>cruzi</i>.</p><p>IC₅₀ values are means of two independent assays, which varied < ±50%.</p><p><i>In vitro</i> activity against <i>T</i>. <i>cruzi</i> in IC₅₀ (μM) of compounds fulfilling hit criteria.</p

In vitro activity against T.b. rhodesiense in IC50 (μM) of compounds fulfilling hit criteria.

<p>^a<i>T</i>. <i>b</i>. <i>rhod</i>.:<i>T</i>. <i>b</i>. <i>rhodesiense</i> strain STIB 900, trypomastigotes.</p><p>^bCytotoxicity on L6 cells.</p><p>^cSelectivity index: IC₅₀ Cytotoxicity L6/ IC₅₀<i>T</i>. <i>b</i>. <i>rhodesiense</i>.</p><p>IC₅₀ values are means of two independent assays, which varied < ±50%.</p><p>In vitro activity against T.b. rhodesiense in IC50 (μM) of compounds fulfilling hit criteria.</p