Map of cutaneous leishmaniasis cases by zone of residence (urban/peri-urban/rural), Santa Catarina, Brazil (n = 452, 2001–2009).

<p>Map of cutaneous leishmaniasis cases by zone of residence (urban/peri-urban/rural), Santa Catarina, Brazil (n = 452, 2001–2009).</p

Comparison of case demographics and clinical characteristics of cutaneous leishmaniasis cases between the northeastern and southern/western regions of Santa Catarina from 2001 to 2009 by chi-squared test or Fisher’s exact test.

<p>Comparison of case demographics and clinical characteristics of cutaneous leishmaniasis cases between the northeastern and southern/western regions of Santa Catarina from 2001 to 2009 by chi-squared test or Fisher’s exact test.</p

Molecular characterization of <i>Leishmania</i> species present in biopsy sample.

<p>(A) PCR amplification products of the <i>Leishmania</i> kDNA minicircle conserved region from cutaneous leishmaniasis cases in Santa Catarina, Brazil, stained with ethidium bromide; Restriction fragment length polymorphism (RFLP) of the amplified the kDNA minicircle conserved region digested with restriction enzymes <i>Hae</i>III (B) and <i>Ava</i>I (C), and analyzed by electrophoresis in 10% polyacrylamide gels with silver staining; Lane 1–2 = <i>L. (V.) braziliensis</i> cases from the northeastern region; Lane 3–4 = <i>L. (V.) braziliensis</i> cases from the southern and western regions, Lane 5–6 = <i>L. (L.) amazonensis</i> cases, Lane 7 = reference strain <i>L. (V.) braziliensis</i> (MHOM/BR/75/M-2904), Lane 8 = reference strain <i>L. (L.) amazonensis</i> (IFLA/BR/67/PH8), NC = negative control, MW = molecular weight.</p

Multivariate logistic regression predicting cutaneous leishmaniasis case relapse in Santa Catarina, Brazil.

*<p>p<0.05,</p>†<p>p<0.001.</p

General characteristics of the <i>T. rangeli</i> genome.

<p>* Excluding proteins of unknown function.</p><p>General characteristics of the <i>T. rangeli</i> genome.</p

Evolutionary history of the Trypanosomatidae family obtained through a phylogenomic approach using (<b>A</b>) the neighbor joining (NJ) or (<b>B</b>) the maximum likelihood (ML) methods.

<p>In the NJ results, the percentage of replicate trees in which the associated taxa clustered together in the bootstrap test (100 replicates) is shown next to the branches. In the ML results, each internal branch indicates, as a percentage, how often the corresponding cluster was found among the 1,000 intermediate trees. The scale bar represents the number of amino acid substitutions per site.</p

Number of gene clusters shared by the <i>T. rangeli</i>, <i>T. cruzi</i>, <i>T. brucei</i> and <i>L. major</i> genomes.

<p>Analyzes were performed using the following genome versions and gene numbers retrieved from the TriTrypDB: <i>Leishmania major</i> Friedlin (V. 7.0/8,400 genes), <i>Trypanosoma brucei</i> TREU927 (V. 5.0/10,574 genes), <i>Trypanosoma cruzi</i> CL Brener Esmeraldo (V. 7.0/10,342 genes) and Non-Esmeraldo (V. 7.0/10,834 genes). A total of 7,613 <i>T. rangeli</i> genes were used. BBH analysis used a cut-off value of 1e-05, positive similarity type and similarity value of 40% following manual trimming for comparison with COG analysis in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003176#pntd.0003176-ElSayed1" target="_blank">[55]</a> generating the numbers in the rectangles.</p

Molecular karyotype of <i>Trypanosoma rangeli</i>.

<p><b>A</b>. Chromosomal bands of Choachí and SC-58 isolates were separated via PFGE and stained with ethidium bromide. The bands were numbered using Arabic numerals, starting from the smallest band. <b>B</b>. Chromosomal bands from <i>T. rangeli</i> (Choachí and SC-58 strains) and <i>T. cruzi</i> (clone CL Brener) were fractioned using a protocol optimized to separate small DNA molecules, revealing the absence of minichromosomes. The brackets represent the size range of <i>T. brucei</i> minichromosomes (30 and 150 kbp).</p

Representation of the telomeric and subtelomeric regions of <i>Trypanosoma rangeli</i>, <i>T. cruzi</i> and <i>T. brucei</i>.

<p>The two types of telomeres identified in <i>T. rangeli</i> and two others representing the heterogeneity of <i>T. cruzi</i> chromosome ends are shown. The size of the subtelomeric region, which extends between the telomeric hexamer repeats and the first internal core genes of the trypanosomes, is indicated below each map. Boxes indicate genes and/or gene arrays. The maps are not to scale. The <i>T. brucei</i> and <i>T. cruzi</i> maps were adapted from <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003176#pntd.0003176-ElSayed1" target="_blank">[55]</a>, <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0003176#pntd.0003176-MoraesBarros1" target="_blank">[98]</a>.</p

The RNAi machinery is not active in <i>Trypanosoma rangeli</i>.

<p>Western blot analysis of eGFP silencing via siRNA in <i>T. rangeli</i> and Vero cells expressing eGFP. For the Western blot assays, anti-GFP and anti-alpha tubulin antibodies were used. In each blot, wild-type cells (1), eGFP cells (2), eGFP cells transfected with Mock siRNA (3), eGFP cells transfected with EGFP-S1 DS Positive Control (IDT)(4) and eGFP cells transfected with eGFP antisense siRNA (5) are shown sequentially. The experiments were performed in biological triplicates.</p