Median Joining Network

Median Joining Networ

NADH1 gene alignment

NADH1 gene alignmen

Inhibitory concentrations (50%) of several antimalarial drugs against <i>P. falciparum</i> 3D7 strain determined by the Hemozoin detection assay at different times of incubation.

<p>Mean inhibitory concentration values (50%) ± one standard deviation are presented.</p><p>Standard deviation values are not shown for results that were not supported by at least three independent experiments. Time-points identified with (*) were not systematically analyzed, since the 24 hour time-point was used as the preferential time-point to reliably calculate IC50 values (as discussed in the manuscript).</p><p>(X) values could not be determined; (n.d.) no data available.</p

Effect of chloroquine on the growth curve of <i>P. falciparum</i> sensitive and resistant strains.

<p>Synchronous cultures of sensitive (3D7, parasitemia of 1.3%) and resistant (Dd2, parasitemia of 1.4%) <i>P. falciparum</i> strains were incubated for 48 hours with doubling concentrations of chloroquine and analyzed at 6 hourly intervals. The inhibitory effect of chloroquine at higher concentrations (>25 nM) is clearly visible (arrow) after 18 hours of incubation (A). The resistant strain can easily be distinguished from the sensitive strain with growth curves of all drug concentrations being identical to the drug free control (B). Each time point represents the mean value of triplicate samples ± one SD.</p

Gating for detection of depolarizing parasitized red blood cells in a <i>Plasmodium falciparum</i> culture.

<p>Flow cytometric analysis of an uninfected and a synchronized <i>P. falciparum</i> (3D7) infected culture (1.5% parasitemia) after 24 hours of incubation, stained with SYBR green I. Plots of forward vs. side scatter for the uninfected and infected cultures appear in Figures A and B; corresponding plots of side scatter vs. depolarized side scatter appear in Figures C and D. The gates in Figures C and D identify the depolarizing events. Figures E and F (see text) illustrate gates defining SYBR green-positive parasitized cells. The blue dots on Figure F represent the depolarizing events. Staining with the red blood cell surface marker (CD235) shows that 99.5% of events in a stained sample (red line) exhibit fluorescence above the highest level measured in an unstained control (black line), indicating that the detected events are red blood cells (G). In the SYBR green I histogram (H) of the infected culture, the overall population (black line) shows a distinct peak with a high fluorescent intensity in the third decade. This peak corresponds mainly to the gated population of depolarizing events (pink line). Because SYBR green I intensity correlates with DNA content and thus with parasite level of maturation, the depolarizing population (pink line) consists mainly (79.4%) of mature parasites. The highly red- and green-fluorescent events visible outside the SYBR green gate just to the right of its apex represent contaminating white blood cells among the donor red cells.</p

Multiple logistic regression analyses to identify risk factors for helminth infections.

†<p>as compared to AZT-3TC-NVP;</p><p>data are odds ratios (95% confidence intervals);</p><p>odds ratios for ARTs are in comparison to therapy with AZT-3TC-NVP;</p><p>odds ratios are not given for variables that had no significant effect and were removed from regression model;</p><p>the following variables did not significantly affect risk for any of the above infections: gestational age, wearing shoes, use of dietary supplements and height;</p>*<p>p<0.05,</p>**<p>p<0.005,</p>***<p>p<0.0005.</p

Effect of artesunate on the growth curve of <i>P. falciparum</i> (3D7) and the effect of 12 hourly renewing of artesunate during incubation.

<p>Synchronous cultures of a <i>P. falciparum</i> 3D7 strain were incubated for 48 hours with doubling concentrations of artesunate (A) or with a single concentration of 8 nM of artesunate for the whole time or renewed at 12 hour intervals (B and C). Figures A and B show detection of Hz (depolarizing events) while Figure C shows detection of SYBR green I fluorescence (DNA in parasites). The inhibitory effect of artesunate was already detectable after 18 hours of incubation (A). Similar to artemisinin (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061606#pone-0061606-g004" target="_blank">Figure 4C</a>), the growth curve of artesunate at 4 nM showed a 6 hourly delayed growth curve from 18 to 42 hours, including a 6 hour delay in the peak, occurring at 36 hours. Interestingly, the growth curve at 8 nM seemed to show inhibition until 30 hours, when a slight increase was observed (A and B). However, renewing artesunate at 12 hourly intervals eliminates this effect (green line in B). The percentage of SYBR green I positive events remained approximately the same during the 48 hours of incubation (C). This indicates that parasites at the non-renewed 8 nM concentration showed some maturation as indicated by Hz detection but were unable to replicate. Each time point represents the mean value of triplicate samples ± one SD.</p

Antimalarial activities of several antimalarial drugs determined by the Hemozoin detection assay and the HRP2 assay.

<p>The averages of 50% inhibitory concentration values ± one standard deviation are presented above.</p>*<p>For the Hemozoin detection assay each drug was tested at least three times (except for the novel compound NITD246).</p><p>HRP2 – Histidine-rich protein 2;</p>1)<p>after 72 hours of incubation.</p