We found similar evidence of effect modification when we restricted the analysis to cases with myelomeningocele, a more severe type of spina bifida. The cutoff for mother’s toenail arsenic concentrations was defined by the median of study population (0.46 µg/g toenail). Serum folate concentrations were higher among mothers who reported folic acid use during pregnancy compared to those who did not report folic acid use (10.20 ng/ml and 7.96 ng/ml, respectively).
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We collected various biomarkers to investigate how arsenic contributes to the risk of spina bifida. Environmental exposure to arsenic is of particular concern and may be a potential contributor to spina bifida risk, especially in areas of the world with high arsenic exposures and micronutrient deficiencies. Folic acid supplement use reduces the risk of spina bifida , and fortification of staple foods with folic acid has been successful in decreasing spina bifida rates in multiple countries 4, 5.
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The study involved a hospital visit that included a physical examination of the child and their parents, the administration of questionnaires, and the collection of biological samples. BFour samples failed Boston Children’s Hospital Research Computing QC due to one sex mismatch or issues with familial relatedness analysis (i.e. two individuals were determined to have wrong family relationship statuses, and one additional child sample was removed due to subsequent unavailability of parent information). Flowchart of saliva DNA sample selection and case/control family member inclusion in the study.
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- We employed logistic regression models to assess the associations between arsenic concentrations and folic acid use (predictors) and case status.
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- We were able to identify novel variants outside of the traditionally explored biological pathways, such as folate metabolism.
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Next, the Hardy-Weinberg Equilibrium test (–hwe 0.05) was performed to also exclude variants. Specifically, genotypes were called against GRCh37/hg19 reference and made available for downstream analysis as VCFv4.2 format files. The GSA chip can accommodate 24 samples and includes an array of 688,032 total markers.
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- The samples were stored and shipped to the Dartmouth Trace Element Analysis Core at room temperature.
- Our final study population included 294 cases and 163 controls.
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We used stratified models to examine the associations between folic acid and spina bifida at different levels of arsenic exposure. Although folic acid-based preventive strategies have been successful in reducing rates of spina bifida, some areas continue to be at higher risk because of chemical exposures. What makes Spinra worth major factor to consider for Bangladeshi gamers is the combination of a game catalogue improved proven software program high quality, a payment framework that mirrors authentic localisation rather than superficial market adjustment, and a mobile experience that stands up regularly throughout the devices and connection types that qualify genuine usage patterns in Bangladesh. Past the welcome package, Spin Ra preserves a advertising schedule that includes reload bonuses on subsequent down payments, totally free spins offers linked to specific port titles and loyalty-based benefits for gamers who use the platform consistently.
A subset of individuals from the case-control study had saliva DNA of sufficient quality and were included in the genetic analyses (Figure 1). Control children were excluded from the main study if the child was diagnosed with cancer or a condition considered to be related to arsenic exposure (i.e. arsenic-related keratosis). Various candidate gene studies of neural tube defects in humans have examined one or a few genes (Finnell et al., 2021), particularly those that are orthologs of risk genes in mice or genes involved in folate pathways (Greene & Copp, 2014). Determinants of neural tube defects include a combination of environmental, nutritional, and genetic risk factors (Greene & Copp, 2014; Harris & Juriloff, 2007). At minimum, more surveillance for spina bifida is needed in high arsenic areas such as Bangladesh.
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Previous work in Bangladesh in a large pregnancy cohort has shown high correlations between pregnant women’s toenail arsenic measurements during the first trimester and 1 month post-partum and also with measurements from their infant’s toenails at age 1 month . We were not able to assess all pregnancies that included neural tube defects because Bangladesh does not have a birth defect surveillance program that collects this information from ultrasonography centers or from all deliveries. We used biomarkers (e.g. toenail arsenic) to minimize this bias. If arsenic exposure was related to these more severe outcomes, our results could represent a downward bias towards or beyond the null . It is possible that even higher arsenic exposures are needed to demonstrate a primary effect. In Folbp2 knockout mice, for example, mice with this specific defect in folate transport had higher rates of NTDs after arsenic exposure, and mice nullizygous for genes encoding proteins in cellular uptake of folate were more susceptible to arsenic-induced NTDs .
