Structural and physico-chemical features of the glycerophospholipids investigated in the present work.

a<p>T_m values were obtained from the literature <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088976#pone.0088976-Silvius1" target="_blank">[67]</a>.</p

Kinetics of heme crystallization promoted by different commercial and biological lipids.

<p>Heme crystallization reactions were induced <i>in vitro</i> mediated by uPC, uPS or uPE (100 µM), a blended phospholipid mixture of commercial uPS (14%), uPC (32%) and uPE (51%) or 10 µg/mL of total lipids isolated from PMVM of <i>R. prolixus</i> previously fed with plasma or blood. Data are expressed as mean ± SD, of at least three different experiments and fitted using the Avrami equation as described in the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088976#s2" target="_blank">methods section</a>. To perform the Avrami analysis, the uPC-induced kinetics were independently analyzed at early and late times, which are shown as insets.</p

Fourier transformed infrared spectroscopy identifies the crystals produced by different lipids as β-hematin.

<p>The crystals were produced by 100 µM uPC (A), uPS (B), uPE (C) or 10 µg/mL total lipids isolated from PMVM of <i>R. prolixus</i> previously fed with plasma (D) or blood (E). The characteristic iron-carboxylate peaks of β-hematin at 1210 and 1663 cm⁻¹ are shown.</p

Unsaturated phosphatidylethanolamine produced homogeneous crystals morphologically similar to those induced by <i>R. prolixus</i> midgut lipids.

<p>Transmission electron microscopy of crystals induced by 100 µM uPC (A), uPS (B) or uPE (C) or 10 µg/mL total lipids isolated from <i>R. prolixus</i> midgut content previously fed with plasma (D) or blood (E). The inset shown in uPC represent a very small population of regularly shaped crystals produced by uPC. Scale bars represent 100 nm for all images, including the inset.</p

Fitted kinetic parameters for β-hematin formation mediated by commercial phospholipids and biological lipids from <i>R. prolixus</i> midgut.

<p>Values were expressed as mean ± SEM of five distinct kinetic parameters obtained from the data shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0088976#pone-0088976-g004" target="_blank">Figure 4</a>. Statistical analyses between groups were performed by using the Mann-Whitney (superscript letters) or Student's t tests (superscript symbols). <i>^ap</i><0.05 relative to uPC; <i>^bp</i><0.01 relative to uPC; <i>^cp</i><0.05 relative to uPS; <i>^dp</i><0.005 relative to uPS; <i>^ep</i><0.005 relative to uPE; <i>^fp</i><0.001 relative to uPE; <i>^gp</i><0.0005 relative to RML plasma. Comparisons of reactions half-lives showed that all groups were statistically distinct among each other (<i>p</i><0.03), with exception of the uPE and Blend comparison (<i>p</i> = 0.88).</p

Bootstrapped phylogram of <i>Rhodnius prolixus</i> midgut lipocalins aligned with their best matches to the NR database.

<p>Bootstrap values above 50% are shown on the branches. The bottom line indicates 20% amino acid sequence divergence between the proteins. <i>R. prolixus</i> sequences are shown by the notation RP followed by a unique number. The remaining sequences, obtained from GenBank, are annotated with the first three letters of the genus name, followed by the first three letters of the species name, followed by their GenBank GI number. One thousand replicates were done for the bootstrap test using the neighbor joining test.</p

Bootstrapped phylogram of <i>Rhodnius prolixus</i> and other aspartyl proteinases.

<p>Bootstrap values above 50% are shown on the branches. The bottom line indicates 10% amino acid sequence divergence between the proteins. <i>R. prolixus</i> sequences are shown by the notation RP followed by a unique number and have a red circle preceding their names. The <i>Triatoma infestans</i> sequences from Balczun et. al. <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0002594#pntd.0002594-Balczun1" target="_blank">[2]</a> have a green marker. The remaining sequences were obtained from GenBank and are annotated with the first three letters of the genus name, followed by the first three letters of the species name, followed by their GenBank GI number. One thousand replicates were done for the bootstrap test using the neighbor joining test.</p

Bootstrapped phylogram of <i>Rhodnius prolixus</i> and other insect peritrophin annotated as Group IV peritrophin in Fig. 1.

<p>Bootstrap values above 50% are shown on the branches. The bottom line indicates 10% amino acid sequence divergence between the proteins. <i>R. prolixus</i> sequences are shown by the notation RP followed by a unique number. The remaining protein sequences were obtained from GenBank and are annotated with the first three letters of the genus name followed by the first three letters of the species name followed by their GenBank GI number. All non-<i>Rhodnius</i> sequences derive mostly from mosquitoes, with one deriving from a flea and another from a sand fly. Roman numerals indicate clades with mixed mosquito genera. Ten thousand replicates were done for the bootstrap test using the neighbor joining method.</p

Functional classification of AM-overexpressed transcripts (>10× compared to posterior) from <i>Rhodnius prolixus</i>.

<p>Functional classification of AM-overexpressed transcripts (>10× compared to posterior) from <i>Rhodnius prolixus</i>.</p

Functional classification of gut-overexpressed transcripts (>10× compared to whole body) from <i>Rhodnius prolixus</i>.

<p>Functional classification of gut-overexpressed transcripts (>10× compared to whole body) from <i>Rhodnius prolixus</i>.</p