Data for the samples collected from the three women found infected with <i>P. vivax</i>.

<p>Days when treatment with artemether-lumefantrine (CoA) or IPTp using sulfadoxine-pyrimethamine (SP) was administered are indicated. Del denoted the day of delivery. Samples for PCR or thick blood smears (TBS) were only available for the weeks indicated on the time axis, and the results are presented above this axis. The parasite species identified by PCR or microscopy are provided, as is the average parasitaemia (parasites per µl of blood) recorded for each species: F = <i>P. falciparum</i>; V = <i>P. vivax</i> (shown in red); O = <i>P. ovale</i>; N = negative sample, and “-“ denotes a missing sample. The patient codes provided are those internally and randomly assigned upon recruitment. Samples from patient C012080 from which the <i>Pvdhfr</i> fragment could be amplified and sequenced are indicated by an asterisk (*).</p

Characteristics of <i>P</i>. <i>ovale</i> samples and species identification by microscopy, nested PCR and Real time PCR.

a<p>The period in months (mo) between the last date in the country endemic for malaria and presentation to the hospital in Italy; NA  =  not available.</p>b<p><i>Plas</i>. sp.  =  <i>Plasmodium</i> species; Pf  =  <i>P. falciparum</i>; Pv  =  <i>P. vivax</i>; Pm  =  <i>P. malariae</i>; Po  =  <i>P. ovale</i>; N  =  negative.</p>c<p>The presence of the species <i>P. falciparum</i>, <i>P. vivax</i> and <i>P. malariae</i> was determined by the classical nested PCR (NP-2002) as previously described <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048033#pone.0048033-Snounou5" target="_blank">[21]</a>, and that of <i>P. ovale</i> (<i>P</i>. <i>ovale curtisi</i> and/or <i>P</i>. <i>ovale wallikeri</i>) was determined by nested PCR using recently described oligonucleotides that amplify both of the two species <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0048033#pone.0048033-Calderaro1" target="_blank">[11]</a>.</p

Genetic diversity within the <i>Plasmodium vivax</i> Reticulocyte binding protein genes.

<p>Note: n = number of sequenced isolates; Size = size of the gene analyzed with the non-coding regions excluded; SNP (n) = number of single nucleotide polymorphisms; NS = non-synonymous substitutions; S = synonymous substitutions.</p

Schematic representation of the <i>Pvrbp2a</i> (A) and <i>Pvrbp2b</i> (B) genes.

<p>In each panel, the location of the fragments cloned is indicated below the gene model. Above the gene model, synonymous mutations are indicated by vertical bars below the horizontal bar that represents the gene, whereas non-synonymous mutations are place above this horizontal bar. SNPs that have been confirmed from two independent PCR amplifications are shown as solid lines, whereas those observed only once are represented by dotted lines. The nature and location of confirmed SNPs are also provided as tables, with the non-synonymous mutations highlighted in olive green.</p

Schematic representation of the <i>Pvrbp2d</i> (A) and <i>Pvrbp3</i> (B) genes.

<p>In each panel, the location of the fragments cloned is indicated below the gene model. Above the gene model, synonymous mutations are indicated by vertical bars below the horizontal bar that represents the gene, whereas non-synonymous mutations are place above this horizontal bar. SNPs that have been confirmed from two independent PCR amplifications are shown as solid lines, whereas those observed only once are represented by dotted lines. The nature and location of the confirmed SNPs are also provided in the tables, with the non-synonymous mutations highlighted in olive green. As these two genes are pseudogenes, the consequence of the SNP, synonymous or non-synonymous, has been predicted assuming that a continuous reading frame.</p

The copy number of <i>Pvrbp2a</i>, <i>Pvrbp2b</i> genes in seven <i>P. vivax</i> isolates (Numbered TK1 to TK7) collected in western Thailand (Tak province).

<p>The copy number of <i>Pvrbp2a</i>, <i>Pvrbp2b</i> genes in seven <i>P. vivax</i> isolates (Numbered TK1 to TK7) collected in western Thailand (Tak province).</p

Influence of polymorphism in the genes for the interleukin (IL)-1 receptor antagonist and IL-1ß on tuberculosis

Several lines of evidence suggest that host genetic factors controlling the immune response influence infection by Mycobacterium tuberculosis. The proinflammatory cytokine interleukin (IL)-1beta and its antagonist, IL-1Ra (IL-1 receptor agonist), are strongly induced by M. tuberculosis and are encoded by polymorphic genes. The induction of both IL-1Ra mRNA and secreted protein by M. tuberculosis in IL-1Ra allele A2-positive (IL-1Ra A2(+)) healthy subjects was 1.9-fold higher than in IL-1Ra A2(-) subjects. The M. tuberculosis-induced expression of mRNA for IL-1beta was higher in subjects of the IL-1beta (+3953) A1(+) haplotype (P = 0.04). The molar ratio of IL-1Ra/IL-1beta induced by M. tuberculosis was markedly higher in IL-1Ra A2(+) individuals (P < 0.05), with minor overlap between the groups, reflecting linkage between the IL-1Ra A2 and IL-1beta (+3953) A2 alleles. In M. tuberculosis-stimulated peripheral blood mononuclear cells, the addition of IL-4 increased IL-1Ra secretion, whereas interferon gamma increased and IL-10 decreased IL-1beta production, indicative of a differential influence on the IL-1Ra/IL-1beta ratio by cytokines. In a study of 114 healthy purified protein derivative-reactive subjects and 89 patients with tuberculosis, the frequency of allelic variants at two positions (-511 and +3953) in the IL-1beta and IL-1Ra genes did not differ between the groups. However, the proinflammatory IL-1Ra A2(-)/IL-1beta (+3953) A1(+) haplotype was unevenly distributed, being more common in patients with tuberculous pleurisy (92%) in comparison with healthy M. tuberculosis-sensitized control subjects or patients with other disease forms (57%, P = 0.028 and 56%, P = 0. 024, respectively). Furthermore, the IL-1Ra A2(+) haplotype was associated with a reduced Mantoux response to purified protein derivative of M. tuberculosis: 60% of tuberculin-nonreactive patients were of this type. Thus, the polymorphism at the IL-1 locus influences the cytokine response and may be a determinant of delayed-type hypersensitivity and disease expression in human tuberculosis

Chromosomal location of the <i>Plasmodium vivax</i> Reticulocyte Binding Protein genes.

<p>Chromosomal location of the <i>Plasmodium vivax</i> Reticulocyte Binding Protein genes.</p

Additional file 1: of The suitability of laboratory-bred Anopheles cracens for the production of Plasmodium vivax sporozoites

Raw data on mosquito’s infections and sporozoites production. This file shows the original data analysed and displayed in the graphs in the present manuscript

Variation of gene expression when <i>P. falciparum</i> isolates were incubated with ECs.

<p>The implication of the seven selected genes in the ability of isolates B and C to induce the lung and brain EC death was represented as a ratio by comparing their RNA levels in co-cultures using the modified culture mediums, where EC apoptosis death was detected. For all experiments the means and standard deviation were calculated from duplicate wells from two biological replicates.</p><p>*p<0.05,</p><p>**p<0.005,</p><p>***p<0.0005.</p><p>NS = Non Significant.</p