Comparison of three molecular methods for the detection and speciation of and -0

<p><b>Copyright information:</b></p><p>Taken from "Comparison of three molecular methods for the detection and speciation of and "</p><p>http://www.malariajournal.com/content/6/1/124</p><p>Malaria Journal 2007;6():124-124.</p><p>Published online 15 Sep 2007</p><p>PMCID:PMC2020467.</p><p></p>2% agarose at 100 v for 1 hour. B. (205-bp) and (120-bp) positive samples after 2round nested PCR, run on 2% agarose at 100 v for 1 hour. C. Real-time PCR melt curves of (74.5) and (78). Image provided by Optical Monitor Software v.3.1

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

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

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

Wet mounted erythrocytes subvitally stained with New Methylene Blue (NMB)

<p>(A). A reticulocyte containing the dark reticular matter is indicated by an arrow. The chemical structure of NMB is inserted at the top right hand corner. Wet mounted erythrocytes subvitally stained with Giemsa (B). Two reticulocytes are indicated by arrows. The chemical structure of Giemsa is inserted at the top right hand corner. Wet mounts of <i>Plasmodium vivax</i> (trophozoite stage) infected erythrocytes subvitally stained with NMB (C) and Giemsa (D). The parasitized red cells are indicated by the arrows. The horizontal scale bar at the bottom right of each photomicrograph represents 10 µm. Linear Regression (E) and Bland-Altman (F) comparison of the percentage of reticulocytes (number of reticulocytes per 1000 erythrocytes) detected by Geimsa and NMB wet mount methodologies.</p

Rheopathologic Consequence of <i>Plasmodium vivax</i> Rosette Formation

<div><p>Malaria parasites dramatically alter the rheological properties of infected red blood cells. In the case of <i>Plasmodium vivax</i>, the parasite rapidly decreases the shear elastic modulus of the invaded RBC, enabling it to avoid splenic clearance. This study highlights correlation between rosette formation and altered membrane deformability of <i>P</i>. <i>vivax</i>-infected erythrocytes, where the rosette-forming infected erythrocytes are significantly more rigid than their non-rosetting counterparts. The adhesion of normocytes to the <i>Pv</i>IRBC is strong (mean binding force of 440pN) resulting in stable rosette formation even under high physiological shear flow stress. Rosetting may contribute to the sequestration of <i>Pv</i>IRBC schizonts in the host microvasculature or spleen.</p></div

Accumulation of CQ_GREEN and ³H-CQ in PfCRT-Dd2 microsomes.

<p>Microsomes were incubated with 10 µM of chemosensitizers before addition of CQ_GREEN or ³H-CQ. Ctrl: negative control; Ver: verapamil; Mtp: methiothepin; Mgl: metergoline; Lop: loperamide; Oct: octoclothepin; Mib: mibefradil; L703: L703,606; Chl: chlorprothixene. *: p<0.05, comparing CQ_GREEN uptake against control. ^‡: p<0.05, comparing ³H-CQ uptake against control. Data presented are means ± S.E.M.; n≥3.</p

CQ_GREEN uptake by resistant-type PfCRT is inhibited by mibefradil in a dose-dependent manner.

<p>Microsomes were preincubated with varying concentrations of the PfCRT inhibitor mibefradil prior to addition of CQ_GREEN. At the highest concentration of 10 µM, mibefradil drastically suppressed CQ_GREEN uptake in PfCRT-Dd2 microsomes but had no significant effect on uptake in PfCRT-3D7 microsomes. *, ***: p<0.05 and p<0.001 respectively, against no mibefradil control (Ctrl). Data presented are means ± S.E.M.; n≥3.</p