Numbers of positive (E+) and negative (E-) estrogen metabolites in urine samples according to the fertility status of the study participants.

<p>Women unable to become pregnant after one year of trial (Self-reported primary infertility - Group 2) and those who had borne fewer children than desired (Self-reported secondary infertility - Group 3).</p><p>OR, odds ratio; CI, confidence interval.</p

Status of fertility among Angolan women who provides urine samples positive (<i>S. haematobium</i> +ve) and negative (<i>S. haematobium</i> -ve) for eggs of <i>Schistosoma haematobium</i>, according to the age of the participants.

a<p>Women had been unable to become pregnant after one year of trial (self-reported primary infertility, Group 2) and those who had borne fewer children than they desired (self-reported secondary infertility, Group 3).</p

Major metabolic pathways in cancer initiation by estrogens.

<p>Adapted from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096774#pone.0096774-Lopes1" target="_blank">[21]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096774#pone.0096774-Feldmeier1" target="_blank">[22]</a>.</p

Numbers of positive (<i>S. haematobium</i> +ve) and negative (<i>S. haematobium</i> -ve) urine samples for eggs of <i>Schistosoma haematobium</i> according to the age of the participants.

<p>Numbers of positive (<i>S. haematobium</i> +ve) and negative (<i>S. haematobium</i> -ve) urine samples for eggs of <i>Schistosoma haematobium</i> according to the age of the participants.</p

Typical spectra of representative estrogen metabolites, which were obtained in a single injection.

<p>The m/z of the molecules and the respective retention time at which they appear are highlighted.</p

Map of Angolan study area.

<p>Adapted from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0096774#pone.0096774-Cavalieri4" target="_blank">[42]</a>.</p

Chemical structures, retention times (RT) and m/z values for the four main components identified by LC–MS analysis of urine of <i>Schistosoma haematobium</i>-infected females.

<p>Atomic mass (m); atomic charge (z).</p

Symptoms and fertility state of 93 female residents of the Bengo region of Angola, an area endemic for schistosomiasis haematobia.

<p>Women unable to become pregnant after one year of trial (self-reported primary infertility - Group 2) and those who had borne fewer children than desired (self-reported secondary infertility - Group 3).</p><p><i>Sh egg</i> +ve, positive for eggs of <i>S. haematobium</i> in urine; <i>Sh egg</i> –ve, negative for eggs of <i>S. haematobium</i> in urine.</p><p>OR, odds ratio; CI, confidence interval.</p

Analysis of exome sequencing data identifies a candidate azoospermia mutation in the case of UPD2.

<p>We performed whole-exome sequencing on the case of UPD2 in an attempt to identify a potential genetic cause for this man's azoospermia. We constructed a scoring method to rank order the exome variants in two dimensions: (i) within the set of variants seen in this single exome, the “Individual Score” and (ii) across a large set of exome sequences, the “Population Score”. For each exome variant, the Individual Score, P_ind,, was constructed by summing normalized predictions of functional impact from 5 commonly used annotation algorithms: PhyloP, PolyPhen2, SIFT, GERP, and LRT. This score was then multiplied by the ploidy of the mutant allele (e.g. 1× for a heterozygous genotype and 2× for a homozygous genotype) creating a final Individual Score ranging from 0–10. We also calculated the Individual Score for all variation in the 1000 genomes Phase I sequencing data. To construct the “Population Score” for each variant in the UPD individual, P_pop, we identified the maximum Individual Score variant in the corresponding gene, P_max, within the 1000 genomes data, and defined P_pop = P_ind−P_max. The purpose of the Population Score is to scale the importance of each Individual Score by the extent of pathogenic variation that exists in the population at each gene. Only sites with minor allele frequencies less than 10% in both the 1000 genomes data and the Exome Variant Server (<a href="http://evs.gs.washington.edu/EVS/" target="_blank">http://evs.gs.washington.edu/EVS/</a>) were considered in the analysis. When examining the joint distribution of P_pop and P_ind for the UPD2 individual, we saw an enrichment of large scores for variants on chromosome 2, as expected. The most extreme variant on both scales was a homozygous nonsense mutation in the gene <i>INHBB</i>, the implications of which we discuss in <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003349#pgen-1003349-g006" target="_blank">Figure 6</a>.</p

Homozygous missense mutation of <i>INHBB</i> identified in the case of UPD2.

<p>(A) We validated this candidate by Sanger sequencing in the UPD2 case and control individuals. Mutant and reference nucleotides are highlighted within the blue box, confirming the homozygous T to C nucleotide change observed at chr2:12,1107,305 bp (hg19) of the UPD2 individual. Grey boxes represent the exons of the gene and the red line indicates the location of the observed mutation within the gene. (B) <i>INHBB</i> encodes for the protein, Inhibin βB, which along with inhibin α and inhibin βA, combine combinatorially to form the inhibins and activins. Each protein expressed by <i>INHA</i>, <i>INHBA</i>, <i>INHBB</i> consists of an N-terminal signal peptide (purple), a propeptide (grey), and a subunit chain (green, red or yellow). The mutation identified here results in a M370T change of the inhibin βB subunit chain (location indicated by a vertical red line throughout the diagram). The various inhibin subunits dimerize via disulfide bonds (locations indicated by black lines between subunits). As the βB subunit participates in multiple complexes with antagonistic functions, the functional consequences of loss-of-function or gain-of-function mutations in this protein may be difficult to predict. (C) The role of inhibins and activins in the hypothalamic-pituitary testicular axis. These complexes have diverse functions in the body, but are most well known for their ability to stimulate and inhibit follicle stimulating hormone (FSH) production, a process critical for spermatogenesis. Blue arrows connect hormones to the cell or gland by which they are secreted. Green arrows indicate stimulatory interactions, and red lines indicate inhibitory interactions.</p