The power calculations used to determine the sample size were based on the objectives of the clinical trial and have been reported elsewhere (13). All participants were included in the current analysis. Variables with skewed distribution (hepcidin, ferritin, CRP, AGP, and fecal calprotectin) were log-transformed. Multilevel linear modeling and linear regression were performed using DataDesk 7.0.2 (Data Description Inc.). All multilevel models included subject (discrete variable and random intercept) and day of study (continuous) as independent variables. Forward selection (acceptance threshold at P < 0.05) was used to determine the variables for inclusion in multivariate analysis of factors predicting hepcidin. None of age, sex, length-for-age, or weight-for-age were significant predictors of hepcidin and hence were not included in the model. The relative strengths of associations between hepcidin and its putative regulators were assessed by the F ratio. Children’s mean hepcidin values adjusted for iron status (UIBC), inflammation (CRP), or both were computed using the estimated cell means command in the linear models package of DataDesk. On the basis of previous stable isotope iron absorption studies in children within the same setting, we used a hepcidin threshold of 5.5 ng/ml to separate iron absorbers from nonabsorbers (12, 16). Note that the Bachem ELISA test used here is the same as that used to determine the absorption threshold (5.5 ng/ml) (16), and longitudinal stability was checked as part of the regular quality control. Children were scored as being infected (yes/no) at the twice weekly fieldworker assessments if the mothers reported any of fever, vomiting, diarrhea, cough, or “other illness” or if the child had an axillary temperature of >37.5°C. Nurse-ascribed AEs were listed as all events or subclassified as fever, diarrhea, respiratory or skin infections, where such events could coexist and were counted separately (e.g., a child with fever and respiratory infection would contribute to each analysis). Malaria contributed only three AEs and hence could not be analyzed.

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