Humoral responses against salivary gland homogenate detected in sera of rabbits.

<p><b>(A)</b> IgG antibody levels. (<b>B)</b> IgM antibody levels. Open circles symbolize the non-exposed rabbit group while closed circles indicate the immunized rabbit groups (Rabbits 1–4). Bars represent the geometric means. Antibody levels are expressed as adjusted optical density measured at 492 nm (aOD₄₉₂).</p

Immunization schedule.

<p><b>(A)</b> BALB/c mice were experimentally exposed to the bites of 150 <i>P</i>. <i>perniciosus</i> during twelve weeks and followed during another twelve weeks. A new sand fly exposure was carried out at week 24 to evaluate a B cell recall. <b>(B)</b> Rabbits were exposed to 500 <i>P</i>. <i>perniciosus</i> bites for eight weeks and followed for a total period of twelve weeks (Rabbits 1 and 2) or exposed for four weeks and followed during seven more months (Rabbits 3 and 4). A new sand fly exposure was performed at week 29 and animals were bled the following week (33).</p

Recognition of <i>P</i>. <i>perniciosus</i> salivary proteins by representative sera of mice and rabbits by WB.

<p>Twenty salivary glands were loaded in each well, separated by SDS-PAGE and either Coomassie-stained or transferred to PVDF membranes. The area of membrane corresponding to each well was cut into 4 strips and incubated separately with mice or rabbit individual sera. <b>(A)</b> WB of <i>P</i>. <i>perniciosus</i> SGH with sera of immunized mice at weeks 4, 6, 8, 10, 12, 16, 18, 20, 22, 24 and 25. (N) Negative serum from non-exposed mice. <b>(B)</b> WB of <i>P</i>. <i>perniciosus</i> SGH with sera of immunized rabbits at weeks 0, 1, 2, 3, 4, 9, 13, 17, 21, 25, 29 and 33. (M) Molecular weight marker Precision Dual Xtra Plus (Bio-Rad).</p

Humoral responses against <i>P</i>. <i>perniciosus</i> salivary gland homogenate and recombinant salivary proteins detected in sera of mice.

<p><b>(A), (B), (C)</b> IgG, IgG1 and IgG2a antibody levels against <i>P</i>. <i>perniciosus</i> SGH. <b>(D), (E), (F)</b> IgG, IgG1 and IgG2a antibody levels against the recombinant apyrase rSP01B. <b>G, H, I)</b> IgG, IgG1 and IgG2a antibody levels against the recombinant D7-related protein rSP04. Open circles symbolize the non-exposed mice group while closed circles indicate the immunized mice group. Bars represent the geometric means. Antibody levels are expressed as adjusted optical density measured at 492 nm (aOD₄₉₂).</p

Decontamination of waterborne chemical pollutants by using atmospheric pressure nonthermal plasma: a review

<div><p>Water pollution abatement is a topic of growing interest worldwide, mainly due to the large number of contaminants that continental waters may contain and the need for a safe wastewater reclamation and reuse in many sectors (agriculture, industry, aquifer recharge, and landscape restoration) and industrial recycling to reduce the end-of-pipe treatment. In this review, different advanced oxidation processes based on atmospheric pressure nonthermal plasma treatment for the removal of organic contaminants from waters are critically reviewed. Factors affecting the removal efficiency and energy yield are reviewed for specific organic contaminants (e.g. VOC, phenols, organic dyes, pharmaceuticals and personal care products, surfactants) and also for conventional water quality parameters (e.g. BOD, COD, TOC, , turbidity). Moreover, effects of operational modes (e.g. type of discharge, reactor configuration, plasma gas), catalytic processes (heterogeneous: TiO₂, NiO, and homogeneous: Cu²⁺, Fe²⁺, ), post-treatment reactions and external oxidant addition (O₃, H₂O₂) are also presented.</p></div

Additional file 1: Table S1. of Phlebotomine sand fly survey in the focus of leishmaniasis in Madrid, Spain (2012–2014): seasonal dynamics, Leishmania infantum infection rates and blood meal preferences

Monthly relation of P-values of P. perniciosus and S. minuta densities through the three survey periods. Results of Kruskal-Wallis and Dunn’s multiple comparison tests. (DOCX 18 kb

A selection of down-regulated genes of known function in Pro-Pper/Pro-Stat.

<p>Only the down-regulated genes of known function discussed in the subsection "Differential gene expression between Pro-Pper and Pro-Stat" are included in this table. The following items are specified for each selected clone: fold change (F ≤ -2); SD; Student’s t-test p-value; expect value in alignments (e-value); clone definition according to mapping outcomes a, b and c; Gene Id. retrieved from the database TriTrypDB [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.ref034" target="_blank">34</a>]; functions annotated in the <i>L</i>. <i>infantum</i> genome sequence; qRT-PCR outcomes. See more detailed information in the <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#sec002" target="_blank">Methods</a> section. <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.s006" target="_blank">S5 Table</a> contains the complete set of down-regulated genes of known function.</p

Statistics of Pro-Pper/Pro-Stat differential gene expression profiles.

<p>Absolute frequencies of genes encoding proteins of known function and hypothetical proteins are provided, as well as type c clones and clones that overlap with more than one gene but that were not determined by qRT-PCR.</p

Sand fly gut dissection, <i>in vitro</i> infectivity and mRNA amplification.

<p>(A) Detail of Pro-Pper (40X) within the anterior thoracic midgut (10X). SV: stomodeal valve. Sand flies of an established colony were dissected to extract the whole guts. After that, the anterior part of the thoracic midgut, behind the SV, was separated in a PBS drop and slightly squeezed with a coverslip. Then, the PBS drop containing Pro-Pper in suspension was recovered, thus avoiding carryover of gut tissue as much as possible. (B) The U937 cell line was differentiated with phorbol esters on 8-well chamber slides and <i>in vitro</i> infected with <i>L</i>. <i>infantum</i> Pro-Pper and Pro-Stat (approximately 5 x 10⁴ promastigotes at a phagocyte:promastigote ratio 1:5 were added). The preparations were fixed and stained with modified Giemsa and 100 cells were randomly counted per replicate. The average number of amastigotes per infected cell was measured at 48 h post-infection. Mean ± SD: 2.7 ± 0.4 in the case of Pro-Stat and 4.8 ± 0.9 in the case of Pro-Pper. (C) Agarose gel electrophoresis of aaRNA samples used for the microarray analysis after synthesis of labeled cDNA. Total RNA was purified from isolated Pro-Pper immediately after dissection and doubly amplified (aaRNA) with MessageAmpII aRNA Amplification Kit (Life Technologies), due to sample amount requirements. Pro-Stat RNA was isolated and processed following the same procedure as for Pro-Pper. Five μl aliquots of the aaRNA samples were run at 5 V/cm in a 1.5% agarose gel prepared with RNase-free water after treatment of the electrophoresis cell, tray and comb with hydrogen peroxide. Three biological replicates of the microarray hybridization experiment were performed.</p

HCL-ST comparison of Pro-Pper with cultured promastigotes and amastigotes of <i>L</i>. <i>infantum</i>.

<p>(A) Pro-Pper vs. Pro-Stat (this study) and Amas [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.ref024" target="_blank">24</a>]. (B) Pro-Stat vs. Pro-Pper (this study), Pro-Log and Amas [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.ref028" target="_blank">28</a>]. A description of clones not found in <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.s005" target="_blank">S4</a>–<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.s009" target="_blank">S8</a> Tables can be found in [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.ref028" target="_blank">28</a>]. All the microarray hybridization experiments were performed by the same procedure and clone nomenclature is equivalent (see Availability of the Supporting Data). For obvious reasons, when more than one gene is represented by a given clone, independent qRT-PCR analysis was performed. This approach allows determining the actual gene that is differentially expressed in each different biological condition (Tables <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.t002" target="_blank">2</a> and <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.t003" target="_blank">3</a>, <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.s005" target="_blank">S4</a> and <a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.s006" target="_blank">S5</a> Tables [<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.ref024" target="_blank">24</a>,<a href="http://www.plosntds.org/article/info:doi/10.1371/journal.pntd.0004693#pntd.0004693.ref028" target="_blank">28</a>]).</p