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Abstract
Fumonisins (FB) are mycotoxins found in maize. They are hypothesised risk factors for neural tube defects (NTDs) in humans living where maize is a dietary staple. In LM/Bc mice, FB1-treatment of pregnant dams induces NTDs and results in increased levels of sphingoid base 1-phosphates in blood and tissues. The increased level of sphingoid base 1-phosphates in blood is a putative biomarker for FB1 inhibition of ceramide synthase in humans. Collection of blood spots on paper from finger sticks is a relatively non-invasive way to obtain blood for biomarker analysis. The objective of this study was to develop and validate in an animal model, and ultimately in humans, a method to estimate the volume of blood collected as blood spots on absorbent paper so as to allow quantification of the molar concentration of sphingoid base 1-phosphates in blood. To accomplish this objective, blood was collected from unexposed male LM/Bc and FB1-exposed pregnant LM/Bc mice and humans and applied to two types of absorbent paper. The sphingoid base 1-phosphates, absorbance at 270 nm (A270), and total protein content (Bradford) were determined in the acetonitrile:water 5% formic acid extracts from the dried blood spots. The results show that in both mouse and human the A270, total protein, and blood volume were closely correlated and the volume of blood spotted was accurately estimated using only the A270 of the extracts. In mouse blood spots, as in tissues and embryos, the FB1-induced changes in sphingolipids were correlated with urinary FB1. The half-life of FB1 in the urine was short (<24 h) and the elevation in sphingoid base 1-phosphates in blood was also short, although more persistent than the urinary FB1.
Acknowledgements
The authors thank Ms. Melinda Vongkunthong for her assistance in mouse model development. We also thank all the women and men who participated in the human studies, the Ministry of Health of Guatemala, the leaders in the communities of Chimaltenango and Escuintla and the leadership of the USDA-ARS South Atlantic Area and Russell Research Center without whose cooperation these studies could not have been performed. The authors also thank Adela Ruiz, Rosa Chovix and Waldemar González for the field work and sample collection in Guatemala, Marta María Méndez, Cecilia de Mayorga, Luis Rodríguez and Flor Díaz for the processing of samples in Guatemala. For all animal studies Institutional Animal Care and Use Committee approved protocols were followed and for human studies Institutional Review Board-approved protocols were followed.