Association analyses between genotypes and serum sulfate, and other phenotypic measures were conducted using a regression-based method that models variation of the trait of interest as a function of measured covariates, measured genotypes and a polygenic component that accounts for phenotypic correlation due to relatedness. SLC13A1 encodes the apical membrane, sodium-sulfate cotransporter NaS1, which is responsible for sulfate re absorption in the intestines and kidneys . The enrichment of these SLC13A1 nonsense variants in the Amish provides us the unique opportunity to dissect the role of sulfate and these sulfate-lowering variants in androgenic hormone homeostasis. Serum sulfate concentration 0. Due to the availability of extensive genealogical records  , virtually all present-day Amish can be linked into a single, generation pedigree. Free testosterone was calculated according to the method of Vermeulen et al. In , Noordam et al.
All analyses included age, age-squared, and gender as covariates. Cultural and religious beliefs have maintained the Amish as distinct from the general population. While paradoxical, these results illustrate the complexity of the mechanisms involved in DHEA homeostasis and warrant additional studies to better understand sulfate's role in hormone physiology. Materials and methods 2. Study population This report is based on the Old Order Amish community living in Lancaster County, PA, whom our research group has been studying since The enrichment of these SLC13A1 nonsense variants in the Amish provides us the unique opportunity to dissect the role of sulfate and these sulfate-lowering variants in androgenic hormone homeostasis. The concentration of PAPS equilibrates rapidly with that of serum sulfate, thus the rate of PAPS synthesis, and consequently the rate of sulfation, are thought to be dependent on endogenous sulfate concentrations  , . In order to improve accuracy, a quadratic least squares fit was used, instead of a linear fit, to generate the standard curve . The average serum sulfate concentration among the non-carriers in this cohort was 0. Serum sulfate concentration was significantly lower in SLC13A1 nonsense variant carriers compared to non-carriers in all cohorts examined in this study Table S3. For each sample, sulfate concentration was calculated as the mean of the duplicate measurements. Given that the Old Order Amish of Lancaster County, PA are a founder population with publicly available pedigree data, depositing individual-level data creates the potential for personal identification of our research participants. Written informed consent was obtained from each participant. Free testosterone was calculated according to the method of Vermeulen et al. R12X and rs, c. As a result, loss-of-function alleles in this gene cause in decreased serum sulfate levels  ,  ,  , . All standards and samples were measured in duplicate. The average serum sulfate concentration among the 59 SLC13A1 nonsense variant carriers in this cohort was 0. In , Noordam et al. SHBG was measured by Immulite assay. Serum sulfate concentration 0. This community was founded by several hundreds of individuals who immigrated to Lancaster County, PA from central Europe during the early 18th century, with the present day Lancaster County Amish community comprised of their descendants . In accordance to our human subject research protocols, we are unable to release individual-level data. All analyses of hormone levels obtained in Amish PAPI Study  ,  or Amish HAPI Study  participants containing serum sulfate as a covariate were limited to subjects for whom serum sulfate was measured using a serum aliquot collected at the time of participation in that study. Association analyses between genotypes and serum sulfate, and other phenotypic measures were conducted using a regression-based method that models variation of the trait of interest as a function of measured covariates, measured genotypes and a polygenic component that accounts for phenotypic correlation due to relatedness.
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