Next-generation sequencing identifies the Danforth's short tail mouse mutation as a retrotransposon insertion affecting Ptf1a expression.

The semidominant Danforth's short tail (Sd) mutation arose spontaneously in the 1920s. The homozygous Sd phenotype includes severe malformations of the axial skeleton with an absent tail, kidney agenesis, anal atresia, and persistent cloaca. The Sd mutant phenotype mirrors features seen in human caudal malformation syndromes including urorectal septum malformation, caudal regression, VACTERL association, and persistent cloaca. The Sd mutation was previously mapped to a 0.9 cM region on mouse chromosome 2qA3. We performed Sanger sequencing of exons and intron/exon boundaries mapping to the Sd critical region and did not identify any mutations. We then performed DNA enrichment/capture followed by next-generation sequencing (NGS) of the critical genomic region. Standard bioinformatic analysis of paired-end sequence data did not reveal any causative mutations. Interrogation of reads that had been discarded because only a single end mapped correctly to the Sd locus identified an early transposon (ETn) retroviral insertion at the Sd locus, located 12.5 kb upstream of the Ptf1a gene. We show that Ptf1a expression is significantly upregulated in Sd mutant embryos at E9.5. The identification of the Sd mutation will lead to improved understanding of the developmental pathways that are misregulated in human caudal malformation syndromes.

Comprehensive genetic analysis of OEIS complex reveals no evidence for a recurrent microdeletion or duplication.

Omphalocele-exstrophy of the bladder-imperforate anus-spinal defects (OEIS) complex, or cloacal exstrophy (EC), is a rare constellation of malformations in humans involving the urogenital, gastrointestinal, and skeletal systems, and less commonly the central nervous system. Although OEIS complex is well-recognized in the clinical setting, there remains a significant lack of understanding of this condition at both the developmental and the genetic level. While most cases are sporadic, familial cases have been reported, suggesting that one or more specific genes may play a significant role in this condition. Several developmental mechanisms have been proposed to explain the etiology of OEIS complex, and it is generally considered to be a defect early in caudal mesoderm development and ventral body wall closure. The goal of this study was to identify genetic aberrations in 13 patients with OEIS/EC using a combination of candidate gene analysis and microarray studies. Analysis of 14 candidate genes in combination with either high resolution SNP or oligonucleotide microarray did not reveal any disease-causing mutations, although novel variants were identified in five patients. To our knowledge, this is the most comprehensive genetic analysis of patients with OEIS complex to date. We conclude that OEIS is a complex disorder from an etiological perspective, likely involving a combination of genetic and environmental predispositions. Based on our data, OEIS complex is unlikely to be caused by a recurrent chromosomal aberration.

Occupational and environmental mercury exposure among small-scale gold miners in the Talensi-Nabdam District of Ghana's Upper East region.

Mercury use in small-scale gold mining is ubiquitous across Ghana but little is known about the extent to which such activities have contaminated community residents and miners. Here, occupational exposures to elemental mercury (via urine sampling) and dietary exposures to methylmercury (via hair sampling) were assessed among 120 participants recruited from a mining community located in the Talensi-Nabdam District of Ghana's Upper East region during summer 2009. More than one-fifth of the participants had moderately high levels of urinary mercury (>10µg/L) and 5% had urine mercury levels that exceeded the WHO guideline value of 50µg/L. When participants were stratified according to occupation, those active in the mining industry had the highest mercury levels. Specifically, individuals that burned amalgam had urine mercury levels (median: 43.8µg/L; mean ± SD: 171.1±296.5µg/L; n=5) significantly higher than median values measured in mechanical operators (11.6µg/L, n=4), concession managers/owners (5.6µg/L, n=11), excavators that blast and chisel ore (4.9µg/L, n=33), individuals that sift and grind crushed ore (2.2µg/L, n=47), support workers (0.5µg/L, n=14), and those with no role in the mining sector (2.5µg/L, n=6). There was a significant positive Spearman correlation between fish consumption and hair mercury levels (r=0.30) but not with urine mercury (r=0.18) though further studies are needed to document which types of fish are consumed as well as portion sizes. Given that 200,000 people in Ghana are involved in the small-scale gold mining industry and that the numbers are expected to grow in Ghana and many other regions of the world, elucidating mercury exposure pathways in such communities is important to help shape policies and behaviors that may minimize health risks.