Multiple novel prostate cancer susceptibility signals identified by fine-mapping of known risk loci among Europeans

Genome-wide association studies (GWAS) have identified numerous common prostate cancer (PrCa) susceptibility loci. We have fine-mapped 64 GWAS regions known at the conclusion of the iCOGS study using large-scale genotyping and imputation in 25 723 PrCa cases and 26 274 controls of European ancestry. We detected evidence for multiple independent signals at 16 regions, 12 of which contained additional newly identified significant associations. A single signal comprising a spectrum of correlated variation was observed at 39 regions; 35 of which are now described by a novel more significantly associated lead SNP, while the originally reported variant remained as the lead SNP only in 4 regions. We also confirmed two association signals in Europeans that had been previously reported only in East-Asian GWAS. Based on statistical evidence and linkage disequilibrium (LD) structure, we have curated and narrowed down the list of the most likely candidate causal variants for each region. Functional annotation using data from ENCODE filtered for PrCa cell lines and eQTL analysis demonstrated significant enrichment for overlap with bio-features within this set. By incorporating the novel risk variants identified here alongside the refined data for existing association signals, we estimate that these loci now explain ∼38.9% of the familial relative risk of PrCa, an 8.9% improvement over the previously reported GWAS tag SNPs. This suggests that a significant fraction of the heritability of PrCa may have been hidden during the discovery phase of GWAS, in particular due to the presence of multiple independent signals within the same regio

A Secreted Factor Coordinates Environmental Quality with Bacillus Development.

Entry into sporulation is governed by the master regulator Spo0A. Spo0A accumulates in its active form, Spo0A-P, as cells enter stationary phase. Prior reports have shown that the acute induction of constitutively active Spo0A during exponential growth does not result in sporulation. However, a subsequent study also found that a gradual increase in Spo0A-P, mediated through artificial expression of the kinase, KinA, during exponential growth, is sufficient to trigger sporulation. We report here that sporulation via KinA induction depends on the presence of an extracellular factor or factors (FacX) that only accumulates to active levels during post-exponential growth. FacX is retained by dialysis with a cutoff smaller than 500 Dalton, can be concentrated, and is susceptible to proteinase K digestion, similar to described quorum-sensing peptides shown to be involved in promoting sporulation. However, unlike previously characterized peptides, FacX activity does not require the Opp or App oligopeptide transporter systems. In addition, FacX activity does not depend on SigH, Spo0A, or ComX. Importantly, we find that in the presence of FacX, B. subtilis can be induced to sporulate following the artificial induction of constitutively active Spo0A. These results indicate that there is no formal requirement for gradual Spo0A-P accumulation and instead support the idea that sporulation requires both sufficient levels of active Spo0A and at least one other signal or condition

KinA-dependent sporulation requires a threshold level of FacX.

<p>MF1913 (P_{<i>hy-spank</i>}-<i>kinA</i>) was grown to exponential phase and 0.2 ml of culture was used to inoculate either fresh CH spiked with 37.5 ul of 67X concentrated conditioned media (1X final concentration)(top) or diluted conditioned media (3 parts conditioned CH to 1 part fresh CH) (bottom). When indicated, <i>kinA</i> expression was induced with IPTG (20 μM). Cells were grown for 2 hrs (top panels) or 2.5 hrs (bottom panels) at 37°C before image capture. Membranes were stained with TMA.</p

FacX is resistant to boiling, but sensitive to Proteinase K.

<p>(A) Conditioned media was boiled for 15 min and utilized in the sporulation assay (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144168#pone.0144168.g002" target="_blank">Fig 2A</a>) with strain MF1913 (P_{<i>hy-spank</i>}-<i>kinA</i>). (B) Conditioned media was treated with proteinase K and utilized in the sporulation assay with strain MF1913 (P_{<i>hy-spank</i>}-<i>kinA</i>). When indicated, KinA was induced with IPTG (20 μM). Cells were grown for 2 hrs at 37°C before image capture. Membranes were stained with TMA.</p

FacX activity is not dependent on Spo0A, SigH, or ComX for production, and does not require the Opp or App peptide uptake systems.

<p>(A) A strain harboring P_{<i>hy-spank</i>}-<i>kinA</i> (MF1913) was grown 2 hrs in the indicated media at 37°C before image capture. (B) A strain containing P_{<i>hy-spank</i>}-<i>kinA</i> in an Δ<i>oppABCDF</i>, Δ<i>appA</i> double mutant (BQA121) was grown 2 hrs at 37°C before image capture. The conditioned media utilized in this experiment was obtained from PY79. When indicated, KinA expression was induced with IPTG (20 μM). Membranes were stained with TMA.</p

<i>Bacillus</i> can be induced to sporulate through expression of a constitutively active allele of Spo0A (Spo0A*) if induced during stationary phase or grown in conditioned media.

<p>(A) A strain harboring P_hy-spank—<i>Spo0A*</i> (MF2146) was grown in CH at 37°C, induced by the addition of IPTG (200 μM) at the indicated optical densities, and grown for 2 hrs at 37°C before image capture. Membranes (white and red) were stained with FM4-64, and DNA (green) was stained with DAPI. (B) A strain harboring P_{<i>hy-spank</i>}-<i>Spo0A*</i> (MF2146) was grown in conditioned media obtained from PY79 as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144168#pone.0144168.g002" target="_blank">Fig 2A</a>. When indicated, Spo0A* was induced by the addition of IPTG (either 200 μM or 500 μM, as indicated, beginning OD₆₀₀ 0.04) and grown for 2 hrs at 37°C before image capture. Membranes were stained with TMA.</p

Media from post-exponential growth contains a protein-based factor that promotes sporulation via KinA induction.

<p>(A) A typical PY79 growth curve in CH medium at 37°. The arrow indicates region in the growth curve (OD₆₀₀ between 1.3 and 1.5) where the conditioned media (CM) was collected. The shaded region indicates the region of the growth curve in which exponential growth occurs. (B) Schematic representation of the KinA-dependent sporulation assay used throughout this study. (C) A strain harboring P_{<i>hy-spank</i>}-<i>kinA</i> (MF1913) was grown as described in (A) with conditioned media collected from PY79. KinA was induced with IPTG (20 μM final concentration). Membranes were stained with TMA.</p

FacX activity is dialyzable.

<p>Conditioned CH medium was dialyzed against fresh CH using dialysis tubing with the indicated molecular weight cutoff. The dialysate was then used to perform the KinA-dependent sporulation assay as shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0144168#pone.0144168.g002" target="_blank">Fig 2A</a> with strain MF1913. When indicated, <i>kinA</i> expression was induced with IPTG (20 μM). Cells were grown for 2 hrs at 37°C before image capture. Membranes were stained with TMA.</p

Family‐based prevention programmes for alcohol use in young people.

The results of this review indicate that there are no clear benefits of family‐based programmes for alcohol use among young people. Patterns differ slightly across outcomes, but overall, the variation, heterogeneity, and number of analyses performed preclude any conclusions about intervention effects. Additional independent studies are required to strengthen the evidence and clarify the marginal effects observed