Fig 1: Immunostaining patterns for Ang-1, Ang-2 and Tie-2 differentiated control from malaria cases. Graphs show percentages of cases positive according to marker, cell components and expression scores. The immunostaining patterns of Ang-1, Ang-2 and Tie-2 in malaria cases were significantly different from those in non-malaria cases, especially the staining for neuronal and astroglial components. However, these proteins were not differentially expressed in CM compared to NCM. (*Logistic Regression with clustered robust standard errors, **Fisher's Exact Test, ***Ordered Logistic Regression with clustered robust standard errors).
Fig 2: Summary of the expression pattern of angiogenic proteins in the brain of fatal malaria patients compared with fatal non-malaria patients. Using immunohistochemistry, increased expression of Ang-1 in neurons and astroglial cells, increased expression of Ang-2 in neurons, and increased expression Tie-2 in vessels were observed in the brain of fatal malaria patients compared to fatal non-malaria patients. Decreased expression of Tie-2 in astroglial cells was observed in fatal malaria cases. Values are odds ratios (brackets = 95% confidence intervals) of higher expression scores (NeuExp, GliExp or VasExp) in malaria compared to control patients.
Fig 3: The immunostaining of Ang-1, Ang-2 and Tie-2 in brain sections from fatal malaria cases (A, B, C, F) and fatal non-malaria cases (D, E). There was heterogeneity in expression patterns of Ang-1, Ang-2 and Tie-2 between patients and within different brain regions in each patient. (A, D) Ang-1 was more commonly seen on larger neurons or astroglial cells, perivascular smooth muscles and pericytes, but not usually on endothelial cells (ECs). Strong staining was occasionally observed on neuropil and axonal bundles. (B, E) Strong Ang-2 labelling on axonal bundles, large neurons, astroglial cells, perivascular smooth muscles, pericytes, ECs and some parasitized red blood cells. (C, F) Strong Tie-2 staining of large and small neurons, astroglial cells, perivascular smooth muscles, pericytes and ECs. (F) A larger vessel showing Tie-2 staining whilst surrounding capillaries, showing significant parasitized erythrocyte sequestration, are completely negative. All sections were counterstained with haematoxylin. Scale bars = 60 µm. Abbreviations: AX = axons, G = astroglial cells, N = neurons, P = parasitized red blood cells, V = vessels.
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