Globin concentration; MCV, mean cell volume; Obs, observations; ROC, PLV-2 POR 8 chemical information receiver operating characteristic; sTfR, soluble transferrin receptor; TfR-F index, transferrin-ferritin index; TIBC, total iron binding capacity. doi:10.1371/journal.pone.0050584.t114 14 24 71 69 35 241 By C reactive protein (CRP): ,12 ng/ml if CRP,1 mg/dl, and ,30 ng/ml if CRP 1 mg/dl. 2 By age: ,50 ng/ml in children 3? months of age, and ,7 ng/ml in children .5 months of age. 3 By CRP: .1.5 if CRP,1 mg/dl, and .0.8 if CRP 1 mg/dl. 4 By age: ,70 fl in children,2 years of age, and ,73 fl in children 2 years of age. Abbreviations: MCHC, mean cell haemoglobin concentration; MCV, mean cell volume; Neg, negative; Pos, positive; sTfR, soluble transferrin receptor; TfR-F index, transferrin-ferritin index; TIBC, total iron binding capacity. doi:10.1371/journal.pone.0050584.tanaemia case-control study but refusing to bone marrow sample donation for the iron biomarkers study here presented. All the explanations were given in Portuguese (the National language) and when required in Changana (the local language). The parentsguardians of all children included in the study provided written informed consent.Study SiteThe study was carried out at the Centro de Investigacao em Sau e de Manhica (CISM) in Manhica District, southern ??Mozambique. The characteristics of the area have been described in detail elsewhere [23,24,25]. Malaria transmission of moderate intensity is perennial with some seasonality. More than 95 of the malaria infections are due to Plasmodium falciparum [26]. Adjacent to the CISM is the Manhica District Hospital (MDH), a 110 bed ?health facility. The main causes of hospital attendance and admission among children in the area are pneumonia [27], malaria [25], anaemia [24], malnutrition and HIV-related diseases (unpublished 23977191 data). HIV prevalence in pregnant women was 29 in 2010 [28].with anaemia (haemoglobin (Hb) ,11 g/dl), and with no history of blood transfusion in the preceding 4 weeks, were recruited as cases if their parents-guardians gave written informed consent. Haemoglobin concentration was measured at the time of recruitment by the HemoCueH system (HemoCueH HB 201+, ?Anghelom, Sweden). A complete clinical examination was performed and the information was entered onto standardized questionnaires together with demographic data. Four ml of venous blood were collected by venipuncture for malaria parasitaemia examination, bacterial culture, full blood count and biochemical and molecular determinations. Participating children were offered voluntary HIV counselling and testing. A bone marrow aspiration was performed from the anterior-superior iliac crest or the tibia, under conscious sedation with parenteral ketamine, atropine and diazepam [29,30,31]. Bone marrow aspirates were not performed in children ,3 months of age or with medical counter-indications such as severe respiratory distress, history of seizures, suspected intracranial hypertension, or any risk at the discretion of the responsible clinician. There were no adverse effects associated to bone marrow biopsy, however there were three adverse effects associated to sedation. One child presented bronchial hypersecretion and bone marrow aspirate was then not performed. Two 
other children vomited during the aspirate, also due to the administration of sedatives. Resuscitation equipment was always available during the procedure. All children received treatment according to their clinical condition and f.Globin concentration; MCV, mean cell volume; Obs, observations; ROC, receiver operating characteristic; sTfR, soluble transferrin receptor; TfR-F index, transferrin-ferritin index; TIBC, total iron binding capacity. doi:10.1371/journal.pone.0050584.t114 14 24 71 69 35 241 By C reactive protein (CRP): ,12 ng/ml if CRP,1 mg/dl, and ,30 ng/ml if CRP 1 mg/dl. 2 By age: ,50 ng/ml in children 3? months of age, and ,7 ng/ml in children .5 months of age. 3 By CRP: .1.5 if CRP,1 mg/dl, and .0.8 if CRP 1 mg/dl. 4 By age: ,70 fl in children,2 years of age, and ,73 fl in children 2 years of age. Abbreviations: MCHC, mean cell haemoglobin concentration; MCV, mean cell volume; Neg, negative; Pos, positive; sTfR, soluble transferrin receptor; TfR-F index, transferrin-ferritin index; TIBC, total iron binding capacity. doi:10.1371/journal.pone.0050584.tanaemia case-control study but refusing to bone marrow sample donation for the iron biomarkers study here presented. All the explanations were given in Portuguese (the National language) and when required in Changana (the local language). The parentsguardians of all children included in the study provided written informed consent.Study SiteThe study was carried out at the Centro de Investigacao em Sau e de Manhica (CISM) in Manhica District, southern ??Mozambique. The characteristics of the area have been described in detail elsewhere [23,24,25]. Malaria transmission of moderate intensity is perennial with some seasonality. More than 95 of the malaria infections are due to Plasmodium falciparum [26]. Adjacent to the CISM is the Manhica District Hospital (MDH), a 110 bed ?health facility. The main causes of hospital attendance and admission among children in the area are pneumonia [27], malaria [25], anaemia [24], malnutrition and HIV-related diseases (unpublished 23977191 data). HIV prevalence in pregnant women was 29 in 2010 [28].with anaemia (haemoglobin (Hb) ,11 g/dl), and with no history of blood transfusion in the preceding 4 weeks, were recruited as cases if their parents-guardians gave written informed consent. Haemoglobin concentration was measured at the time of recruitment by the HemoCueH system (HemoCueH HB 201+, ?Anghelom, Sweden). A complete clinical examination was performed and the information was entered onto standardized questionnaires together with demographic data. Four ml of venous blood were collected by venipuncture for malaria parasitaemia examination, bacterial culture, full blood count and biochemical and molecular determinations. Participating children were offered voluntary HIV counselling and testing. A bone marrow aspiration was performed from the anterior-superior iliac crest or the tibia, under conscious sedation with parenteral ketamine, atropine and diazepam [29,30,31]. Bone marrow aspirates were not performed in children ,3 months of age or with medical counter-indications such as severe respiratory distress, history of seizures, suspected intracranial hypertension, or any risk at the discretion of the responsible clinician. There were no adverse effects associated to bone marrow biopsy, however there were three adverse effects associated to sedation. One child presented bronchial hypersecretion and bone marrow aspirate was then not performed. Two other children vomited during the aspirate, also due to the administration of sedatives. Resuscitation equipment was always available during 
the procedure. All children received treatment according to their clinical condition and f.
