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Fig 1.

Current diagnosis of HAT.

CATT: card agglutination test for trypanosomiasis. CSF: cerebrospinal fluid. RDT: Rapid Diagnostic Test.

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Table 1.

Characteristics of the studied subjects

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Fig 2.

Variation in metabolic coverage of each biofluid.

The percentage shows the number of metabolites detected in each metabolic class (based on matches to the IDEOM database) as a percentage of the total number of metabolites detected in all three groups. Unannotated peaks: masses with no IDEOM database match, no annotated pathway: metabolites that did not match databases for known metabolic pathways. Medium components are those that are commonly found in trypanosome growth medium.

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Fig 3.

Variability across urine samples.

(A) Total ion chromatograms (TICs) show the range in concentration of the ions between samples. Ions are analysed in positive (top) and negative (bottom) ionisation modes. The control group is shown as an example. (B) Principal components analysis shows a lack of separation of the sample groups in raw data.

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Fig 4.

Separation between the metabolite patterns of stage 1 and advanced stage 2 CSF samples.

(A) Principal components analysis plot. (B) Histograms for metabolites showing significant differences between control (C), stage 1 (S1) and advanced stage 2 (S2) infected patients. * indicates p<0.05 compared to stage 1, ** indicates p<0.001 compared to stage 1 in a Students t-test. Bars show standard error of the mean. Relative intensities measure peak areas.

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Fig 5.

Patients can be classified into stage 1 and advanced stage 2 groups using eleven biomarkers in CSF.

O-acetylcarnitine and tryptophan match to authentic standards. Some masses did not match to metabolites in the IDEOM [32] database and are identified by the mass only. Red shading indicates peak area intensities above/below the cut-off for advanced stage 2 disease. More information on the cut-offs are shown in S4 Table.

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Fig 6.

Sleep-inducing metabolites linoleamide and oleamide were increased in both stage 1 and advanced stage 2 patients.

Bars show the mean and the standard error of the mean. Metabolite annotations are by mass only. Relative intensities measure peak areas. * indicates p<0.05 compared to control, ** indicates p<0.001 compared to control in a Student t-test.

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Fig 7.

Metabolite differences in plasma are small, but significant.

(A) Principal components analysis. (B) Extracted peaks for m/z 133 (ornithine) and m/z 216 (aminododecanoic acid). Stage 1: green, advanced stage 2: blue. (C) Histograms for m/z 133 and m/z 216 (relative intensities measure peak areas). ** indicates a p-value of <0.001 in a Students’ t-test. (D) ROC curve for m/z 133 and m/z 216 showing the 95% confidence intervals.

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