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Congenital anomalies are already the second cause of infant mortality in Brazil, as in many other middle-income countries in Latin America. Birth defects are a result of both genetic and environmental factors, but a multifactorial etiology has been more frequently observed. Here, we address the environmental causes of birth defects – or teratogens – as a public health issue and present their mechanisms of action, categories and their respective maternal-fetal deleterious effects. We also present a survey from 2008 to 2013 of Brazilian cases involving congenital anomalies (annual average of 20,205), fetal deaths (annual average of 1,530), infant hospitalizations (annual average of 82,452), number of deaths of hospitalized infants (annual average of 2,175), and the average cost of hospitalizations (annual cost of $7,758). Moreover, we report on Brazilian cases of teratogenesis due to the recent Zika virus infection, and to the use of misoprostol, thalidomide, alcohol and illicit drugs. Special attention has been given to the Zika virus infection, now proven to be responsible for the microcephaly outbreak in Brazil, with 8,039 cases under investigation (from October 2015 to June 2016). From those cases, 1,616 were confirmed and 324 deaths occurred due to microcephaly complications or alterations on the central nervous system. Congenital anomalies impact life quality and raise costs in specialized care, justifying the classification of teratogens as a public health issue.

Paper-based devices are a portable, user-friendly, and affordable technology that is one of the best analytical tools for inexpensive diagnostic devices. Three-dimensional microfluidic paper-based analytical devices (3D-μPADs) are an evolution of single layer devices and they permit effective sample dispersion, individual layer treatment, and multiplex analytical assays. Here, we present the rational design of a wax-printed 3D-μPAD that enables more homogeneous permeation of fluids along the cellulose matrix than other existing designs in the literature. Moreover, we show the importance of the rational design of channels on these devices using glucose oxidase, peroxidase, and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) reactions. We present an alternative method for layer stacking using a magnetic apparatus, which facilitates fluidic dispersion and improves the reproducibility of tests performed on 3D-μPADs. We also provide the optimized designs for printing, facilitating further studies using 3D-μPADs.

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Paper-based devices are an excellent match for low-cost point-of-care testing (POCT) tools. Their userfriendliness, portability, and short time of analysis, coupled with ease of local manufacture make these devices the best option for inexpensive diagnostic testing tools. However, despite all their positive features, these low-cost diagnostic devices must present good performance indicators, such as sensitivity, specificity, and accuracy. We developed and validated a paper-based ELISA for toxoplasmosis diagnosis through the detection of Toxoplasma gondii immunoglobulin G (IgG) antibodies in 100 human serum samples. From among the different ways to define the cut-off value, we chose Youden's J index (cut-off ¼ 21.73 A.U.), which presented a higher sensitivity value. Our paper-based assay presented a sensitivity of 0.96, a specificity of 0.87, and a gray zone comprising 16 samples (AE15% of the cut-off value, with 3 false positive outputs). The accuracy of the test was estimated by using ROC curves (AUC ¼ 0.97). We also created a macro in Microsoft Excel® to estimate the accuracy of the test (m-Accuracy) based on a non-parametric method, which evidenced a value ¼ 0.88, which classifies our test as moderately to highly accurate. We also provide the m-Accuracy macro for download and the paperbased microplate designs for printing, in order to collaborate with the scientific community and facilitate further studies using this platform. The improvement of these diagnostic tools can bring this technology for those who need it, contributing to population health and well-being.

Sarcosine is an amino acid that has been listed as a new indicator for prostate cancer.

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