Microvascular reactivity, endothelial function and biopterin metabolite values among patient groups.

<p>Median and IQR unless otherwise stated. All pterin measurements are μmol/mmol creatinine</p><p>*p<0.05 (for ANOVA, Kruskal-Wallis or χ² test comparing severe malaria, moderately-severe malaria, severe sepsis and healthy controls)</p><p>†Pairwise comparisons between each group and severe malaria using Wilcoxon-Mann-Whitney test: † control vs severe malaria;</p><p>†† moderately severe malaria vs severe malaria;</p><p>††† severe sepsis vs severe malaria.</p><p>Microvascular reactivity, endothelial function and biopterin metabolite values among patient groups.</p

Urinary BH4, BH2 concentrations, BH4/BH2 ratios and Biopterin concentration in each group on enrollment (Kruskal-Wallis: p<0.001). Horizontal bars represent pairwise comparisons between disease groups.

<p>A. Urine BH4 (μmol/mmol creatinine) concentrations in each group on enrollment. Horizontal lines indicate median values for each group. B. Urine BH2 (μmol/mmol creatinine) concentrations in each group on enrollment. Horizontal lines indicate median values for each group. C. BH4/BH2 ratio in each group on enrollment. Horizontal lines indicate median values for each group. D. Urine Biopterin (μmol/mmol creatinine) concentrations in each group on enrollment. Horizontal lines indicate median values for each group. Horizontal bars represent pairwise comparisons between disease groups.</p

Bivariate and multivariate logistic regression analysis of factors associated with increased odds of cerebral malaria.

<p>CI = confidence intervals</p><p>BH₄ = tetrahydrobiopterin</p><p>BH₂ = dihydrobiopterin</p><p>PfHRP-2 = <i>P</i>. <i>falciparum</i> histidine-rich protein-2</p><p>Note: Phenylalanine, Urine BH₄, Urine BH₂ and PfHRP-2 were log-transformed to enable regression analysis. Number of observations included in multivariate model was 96.</p><p>A = included in multivariate logistic regression analysis, but not retained in the final model due to p-value > 0.05 in multivariate analysis.</p><p>B = not included in multivariate logistic regression analysis because of non-significance in bivariate logistic regression analysis.</p><p>Bivariate and multivariate logistic regression analysis of factors associated with increased odds of cerebral malaria.</p

Primers and probes for GTP-cyclohydrolase I mRNA reverse transcriptase polymerase chain reaction.

<p>Primers and probes for GTP-cyclohydrolase I mRNA reverse transcriptase polymerase chain reaction.</p

Baseline characteristics of the 4 clinical groups.

<p>Data mean (95% confidence interval values) unless otherwise indicated.</p><p>ANOVA with Bonferroni post-estimate unless otherwise indicated.</p><p>A = Chi-squared,</p><p>B = Wilcoxon Rank Sum.</p><p>PfHRP-2 = <i>Plasmodium falciparum</i> histidine-rich protein-2.</p><p>N/D = Test not done.</p><p>C = We were unable to obtain measurements of all analytes for all patients enrolled.</p><p>Glucose measurements were obtained in 27, 48 and 43 of the UM, CM and NMC children, respectively.</p><p>Bicarbonate results were obtained 31, 48 and 47 of the UM, CM, and NMC children, respectively. Creatinine results were obtained in 16, 39 and 40 of the UM, CM and NMC children, respectively.</p><p>^D = Three children (1 HC, 1 UM and 1 CM) with bacteriuria remained in the study as the urine was collected by bag collection (i.e., not collected via sterile, invasive procedures), and there was no clinical suspicion of urinary tract infection in any of the three subjects.</p><p>Baseline characteristics of the 4 clinical groups.</p

Study flow diagram.

<p>Screening, enrollment and <i>post-hoc</i> exclusions for the four clinical groups are depicted. CNS = central nervous system; BCS = Blantyre Coma Score; RDT = rapid diagnostic test for <i>P</i>. <i>falciparum</i> (ParacheckPf, Omega Diagnostics); LP = lumbar puncture; SAE = serious adverse event; CSF = cerebrospinal fluid.</p

Urine biopterin concentrations at enrollment in the 4 clinical groups.

<p><b>A</b>: Comparison of total urine biopterins (tetrahydrobiopterin [BH4] + dihydrobiopterin [BH2] + biopterin [B0]) concentrations at enrollment for healthy controls (HC, n = 111), uncomplicated malaria (UM, n = 55), cerebral malaria (CM, n = 45) and non-malaria central nervous system conditions (NMC, n = 48) (p<0.001 by Kruskal-Wallis). <b>B</b>: urine tetrahydrobiopterin (BH₄) concentrations at enrollment for all 4 groups (p<0.001 by Kruskal-Wallis). Central line indicates median. Upper and lower lines indicate inter-quartile range. <b>C</b>: Comparison of urine tetrahydrobiopterin to dihydrobiopterin (BH4:BH2) ratio at enrollment for all 4 groups (p<0.001 by Kruskal-Wallis). The number of subjects with a sample measured in <b>[B]</b> and <b>[C]</b> is the same as Noted above in <b>[A]</b>. The central line indicates median. Upper and lower lines indicate interquartile range.</p

Urine pterins and plasma phenylalanine measurements at enrollment in the 4 clinical groups.

<p>Units: urine pterins μmol/mmol creatinine; amino acid μM.</p><p>Median and inter-quartile ranges, p-value Kruskal-Wallis comparison across all 4 groups. BH₄ = tetrahydrobiopterin, BH₂ = dihydrobiopterin, B₀ = biopterin, NH₂ = dihydroneopterin, N₀ = neopterin.</p><p>A = p-value <0.05 by Wilcoxon Rank Sum comparison between cerebral malaria and healthy controls</p><p>B = p-value < 0.05 by Wilcoxon Rank Sum comparison between cerebral malaria and non-malaria CNS condition</p><p>C = p-value < 0.05 by Wilcoxon Rank Sum comparison between cerebral malaria and uncomplicated malaria</p><p>D = Number of plasma samples for healthy controls, uncomplicated malaria, cerebral malaria and non-malaria CNS conditions was 109, 61, 50, and 51, respectively.</p><p>Urine pterins and plasma phenylalanine measurements at enrollment in the 4 clinical groups.</p

Plasma angiopoietin-2 concentrations at enrollment in the 4 clinical groups.

<p>Comparison of plasma angiopoietin-2 (Ang-2) concentrations at enrollment in a subset of healthy controls (HC, n = 9), uncomplicated malaria (UM, n = 10), cerebral malaria (CM, n = 16) and non-malaria central nervous system conditions (NMC, n = 9) (p<0.001 by Kruskal-Wallis). The central line indicates median. Upper and lower lines indicate inter-quartile range.</p

Urine neopterin concentrations at enrollment in the 4 clinical groups.

<p><b>A</b>: Comparison of total urine neopterins, dihydroneopterin plus neopterin (NH₂ + N₀), concentrations at enrollment for healthy controls (HC, n = 111), uncomplicated malaria (UM, n = 55), cerebral malaria (CM, n = 45) and non-malaria central nervous system conditions (NMC, n = 48) (p<0.001 by Kruskal-Wallis). The central line indicates median. Upper and lower lines indicate inter-quartile range. <b>B</b>: Comparison of urine dihydroneopterin to neopterin (NH₂:N₀) at enrollment for all 4 groups (p = 0.018 by Kruskal-Wallis). The number of subjects with a sample measured is the same as noted above in [A]. The central line indicates median. Upper and lower lines indicate inter-quartile range.</p