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Additional file 2: Table S1. Clinical characteristic of children with cerebral malaria versus the healthy controls in the second paediatric cohort recruited between 2013 and 2016.

Additional file 1: Figure S1. Gating strategy for cytokine producing monocytes: Whole blood samples were stimulated with LPS, labelled with CD14 APC, lysed with 2.0ml of 1 x FACS lysing solution and fixed with BFA before the labelling with Isotype Control (PE) and various cytokine antibodies (PE). Flow cytometer dot plots illustrating the side scatter plot versus CD14 (A) with R1 gate for CD14+ cells (monocytes), the Isotype Control plot for setting the gates (B), IL-6 producing monocytes (CD14+IL-6+ cells) (C) and TNF-α producing monocytes (CD14+TNF-α+ cells) (D).Figure. S2 Gating strategy for monocytes expressing HLA-DR and CD86: Whole blood samples were stimulated with LPS, labelled with CD14 APC, HLA-DR-FITC and CD86-FITC and incubated for 20 minutes. The samples were then lysed with 2.0ml of 1 x FACS lysing solution and washed with PBS before acquisition on Flow Cytometer. The dot plots (A) illustrate the side scatter plot versus CD14 (A) with R1 gate for CD14+ cells (monocytes), geometric mean florescence intensity (GMFI) of HLA-DR (B) and CD86 (C) expression on monocytes from children presenting with different malaria clinical types Figure. S3 Gating strategy for cytokine producing T cells: Whole blood samples were stimulated with PMA+ION, labelled with CD3 PerCP, lysed with 2.0ml of 1 x FACS lysing solution and fixed with BFA before the labelling with Isotype Control (PE) and various cytokine antibodies (PE). The Flow cytometer dot plots illustrate the side scatter plot versus CD3-PerCP (A) with R1 gate for CD3+ lymphocytes (Total T cells), the Isotype Control plot for setting the gates (B), INF-γproducing cells (CD3+IFN-γ+ cells) (C) and TNF producing T cells (CD3+TNF+ cells) (D).

Additional file 1.

Presently, it is difficult to accurately diagnose sepsis, a common cause of childhood death in sub-Saharan Africa, in malaria-endemic areas, given the clinical and pathophysiological overlap between malarial and non-malarial sepsis. Host biomarkers can distinguish sepsis from uncomplicated fever, but are often abnormal in malaria in the absence of sepsis. To identify biomarkers that predict sepsis in a malaria-endemic setting, we retrospectively analyzed data and sera from a case-control study of febrile Malawian children (aged 6-60 months) with and without malaria who presented to a community health center in Blantyre (January-August 2016). We characterized biomarkers for 29 children with uncomplicated malaria without sepsis, 25 without malaria or sepsis, 17 with malaria and sepsis, and 16 without malaria but with sepsis. Sepsis was defined using systemic inflammatory response criteria; biomarkers (interleukin-6 [IL-6], tumor necrosis factor receptor-1, interleukin-1 β [IL-1β], interleukin-10 [IL-10], von Willebrand factor antigen-2, intercellular adhesion molecule-1, and angiopoietin-2 [Ang-2]) were measured with multiplex magnetic bead assays. IL-6, IL-1β, and IL-10 were elevated, and Ang-2 was decreased in children with malaria compared with non-malarial fever. Children with non-malarial sepsis had greatly increased IL-1β compared with the other subgroups. IL-1β best predicted sepsis, with an area under the receiver operating characteristic (AUROC) of 0.71 (95% CI: 0.57-0.85); a combined biomarker-clinical characteristics model improved prediction (AUROC of 0.77, 95% CI: 0.67-0.85). We identified a distinct biomarker profile for non-malarial sepsis and developed a sepsis prediction model. Additional clinical and biological data are necessary to further explore sepsis pathophysiology in malaria-endemic regions.

Alinane U Munyenyembe,1 Kamunkhwala Gausi,1 Tonney S Nyirenda,2 Jasmin Hiestand,3 Jane Mallewa,3 Wilson L Mandala1,4 1Malawi-Liverpool Wellcome Trust, Malaria Immunology Department, Blantyre, Malawi; 2Pathology Department, College of Medicine, Blantyre, Malawi; 3Medicine Department, College of Medicine, Blantyre, Malawi; 4Academy of Medical Sciences, Biological Sciences Department, Malawi University of Science & Technology, Thyolo, Malawi Aim: Although malaria and HIV infections independently affect the electrolyte and hematologic profiles, little is known of how these profiles are affected in individuals coinfected with malaria and HIV. We therefore conducted this study to investigate the electrolyte and hematologic profiles of Malawian adults presenting with either uncomplicated malaria (UM), severe malaria (SM), and those presenting with HIV and UM or HIV and SM.Methods: Study participants were recruited at Queen Elizabeth Central Hospital, and malaria infection was confirmed by rapid diagnostic test and malaria slides, and full blood count, HIV, and wet chemistries were analyzed.Results: Sodium, potassium, calcium, and chloride levels of all 4 study groups were similar to those of healthy controls. Both HIV-infected groups (UM and SM) had lower red blood cell counts and lower hemoglobin concentration than the reference range. Platelet counts were lower in both HIV-uninfected SM cases (64×109/L) and in the HIV-infected SM cases (66×109/L) compared to the reference range (115–290×109/L). HIV– UM cases had higher proportion and absolute counts of neutrophils and white blood cell counts compared to the HIV+ UM cases.Conclusion: HIV infection did not affect the electrolyte profile of Malawian adults presenting with UM or SM but had an effect on red blood cells, Hb concentration, neutrophils, and platelet counts. Keywords: Plasmodium falciparum malaria, HIV infection, electrolytes, hematologic values, HIV-malaria co-infection, Malawian Adults

Typhoid fever is endemic across sub-Saharan Africa. However, estimates of the burden of typhoid are undermined by insufficient blood volumes and lack of sensitivity of blood culture. Here, we aimed to address this limitation by exploiting pre-enrichment culture followed by PCR, alongside routine blood culture to improve typhoid case detection. We carried out a prospective diagnostic cohort study and enrolled children (aged 0-4 years) with non-specific febrile disease admitted to a tertiary hospital in Blantyre, Malawi from August 2014 to July 2016. Blood was collected for culture (BC) and real-time PCR after a pre-enrichment culture in tryptone soy broth and ox-bile. DNA was subjected to PCR for invA (Pan-Salmonella), staG (S. Typhi), and fliC (S. Typhimurium) genes. A positive PCR was defined as invA plus either staG or fliC (CT

Malaria and anaemia are key underlying factors for iNTS disease in African children. Knowledge of clinical and epidemiological risk-factors for iNTS disease has not been paralleled by an in-depth knowledge of the immunobiology of the disease. Herein, we review human and animal studies on mechanisms of increased susceptibility to iNTS in children.