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      Full blood count and haemozoin-containing leukocytes in children with malaria: diagnostic value and association with disease severity

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          Abstract

          Background

          Diligent and correct laboratory diagnosis and up-front identification of risk factors for progression to severe disease are the basis for optimal management of malaria.

          Methods

          Febrile children presenting to the Medical Research Unit at the Albert Schweitzer Hospital (HAS) in Lambaréné, Gabon, were assessed for malaria. Giemsa-stained thick films for qualitative and quantitative diagnosis and enumeration of malaria pigment, or haemozoin (Hz)-containing leukocytes (PCL) were performed, and full blood counts (FBC) were generated with a Cell Dyn 3000 ® instrument.

          Results

          Compared to standard light microscopy of Giemsa-stained thick films, diagnosis by platelet count only, by malaria pigment-containing monocytes (PCM) only, or by pigment-containing granulocytes (PCN) only yielded sensitivities/specificities of 92%/93%; 96%/96%; and 85%/96%, respectively. The platelet count was significantly lower in children with malaria compared to those without (p < 0.001), and values showed little overlap between groups. Compared to microscopy, scatter flow cytometry as applied in the Cell-Dyn 3000 ® instrument detected significantly more patients with PCL (p < 0.01). Both PCM and PCN numbers were higher in severe versus non-severe malaria yet reached statistical significance only for PCN (p < 0.0001; PCM: p = 0.14). Of note was the presence of another, so far ill-defined pigment-containing group of phagocytic cells, identified by laser-flow cytometry as lymphocyte-like gated events, and predominantly found in children with malaria-associated anaemia.

          Conclusion

          In the age group examined in the Lambaréné area, platelets are an excellent adjuvant tool to diagnose malaria. Pigment-containing leukocytes (PCL) are more readily detected by automated scatter flow cytometry than by microscopy. Automated Hz detection by an instrument as used here is a reliable diagnostic tool and correlates with disease severity. However, clinical usefulness as a prognostic tool is limited due to an overlap of PCL numbers recorded in severe versus non-severe malaria. However, this is possibly because of the instrument detection algorithm was not geared towards this task, and data lost during processing; and thus adjusting the instrument's algorithm may allow to establish a meaningful cut-off value.

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          Most cited references38

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          Vertical transmission of HIV-1 infection.

          M-L Newell (2015)
          Vertical transmission is the dominant mode of acquisition of infection for HIV infection in children, and about 1600 infants are newly infected each day worldwide. Without interventions the risk of transmission is between 15% and 35%, and associated with maternal disease progression, prematurity, duration of rupture of membranes, length of labour, and vaginal delivery. Breastfeeding approximately doubles the risk of vertical transmission; the additional risk of transmission through breastfeeding is approximately 15-20%, with about one-third of this accounted for by late postnatal transmission after 3 months of age. Current strategies to reduce the risk of transmission include a short course of anti-retroviral therapy, avoidance of breastfeeding and Caesarean section delivery. However, even if interventions late in pregnancy or around the time of delivery are highly effective in preventing perinatal infection, it is likely that as a public health policy they are of interest only if alternatives to breastfeeding are feasible, affordable, safe and available.
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            Suppression of erythropoiesis in malarial anemia is associated with hemozoin in vitro and in vivo.

            Malarial anemia is a global public health problem and is characterized by a low reticulocyte response in the presence of life-threatening hemolysis. Although cytokines, in particular tumor necrosis factor-alpha (TNF-alpha), can suppress erythropoiesis, the grossly abnormal bone marrow morphology indicates that other factors may contribute to ineffective erythropoiesis. We hypothesized that the cytotoxic hemozoin (Hz) residues from digested hemoglobin (Hb) significantly contribute to abnormal erythropoiesis. Here, we show that not only isolated Hz, but also delipidated Hz, inhibits erythroid development in vitro in the absence of TNF-alpha. However, when added to cultures, TNF-alpha synergizes with Hz to inhibit erythropoiesis. Furthermore, we show that, in children with malarial anemia, the proportion of circulating monocytes containing Hz is associated with anemia (P < .001) and reticulocyte suppression (P = .009), and that this is independent of the level of circulating cytokines, including TNF-alpha. Plasma Hz is also associated with anemia (P < .001) and reticulocyte suppression (P = .02). Finally, histologic examination of the bone marrow of children who have died from malaria shows that pigmented erythroid and myeloid precursors are associated with the degree of abnormal erythroid development. Taken together, these observations provide compelling evidence for inhibition of erythropoiesis by Hz.
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              Intermittent preventive treatment against malaria in infants in Gabon--a randomized, double-blind, placebo-controlled trial.

              Intermittent preventive treatment aims to maximize the protective effects of malaria chemoprophylaxis while minimizing the deleterious effects. In Gabon, 1189 infants received either sulfadoxine-pyrimethamine (SP; 250 and 12.5 mg, respectively) or placebo at 3, 9, and 15 months of age. Children were actively followed-up until 18 months of age. In the intention-to-treat population at 18 months of follow-up, 84 children (17%) in the SP group had > or =1 episode of anemia, versus 108 (21%) in the placebo group (protective efficacy, 22% [95% confidence interval {CI}, -1% to 40%]; P=.06). In the intervention group, there were 66 episodes during 485 person-years at risk, compared with 79 episodes during 497 years in the placebo group (protective efficacy, 17% [95% CI, -24% to 45%; P=.36). The effects were similar at 12 months of follow-up. The study drug was safe and well tolerated. The intervention was efficacious, producing a reduction in risk for anemia but a smaller effect against malaria. It is a valuable additional tool to control malaria in a highly vulnerable age group. Remaining important questions are currently being addressed in further studies. ClinicalTrials.gov identifier: NCT00167843.
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                Author and article information

                Journal
                Malar J
                Malaria Journal
                BioMed Central
                1475-2875
                2008
                12 June 2008
                : 7
                : 109
                Affiliations
                [1 ]Medical Research Unit, Albert Schweitzer Hospital, Lambaréné, Gabon
                [2 ]Institute of Molecular Medicine, Lisbon Medical College, Lisbon, Portugal
                [3 ]Department of Parasitology, Institute of Tropical Medicine, University of Tübingen, Tübingen, Germany
                [4 ]Infectious Diseases Unit, Infectious Diseases Unit, Division of Clinical Microbiology and Infectious Diseases, National Health Laboratory Service and School of Pathology, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg 2193, 7 York Road, Parktown, South Africa
                Article
                1475-2875-7-109
                10.1186/1475-2875-7-109
                2435542
                18549498
                d921dc3a-3b9c-41bd-a0db-ed929be12e1f
                Copyright © 2008 Hänscheid et al; licensee BioMed Central Ltd.

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 21 December 2007
                : 12 June 2008
                Categories
                Research

                Infectious disease & Microbiology
                Infectious disease & Microbiology

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