Demographics and enhanced surveillance.

<p>* Excludes new entrants & visitors</p><p>Demographics and enhanced surveillance.</p

Patients readmitted with infection.

<p>Patients readmitted with infection.</p

Clinical symptoms and signs on admission for 95 patients.

<p>Clinical symptoms and signs on admission for 95 patients.</p

RPS19, RPS21, and RPS24 protein expression using immunofluorescence.

<p>Representative images of the co-expression of RPS19 (Cy3- Red), RPS21 (FITC- Green), and RPS24 (Cy5- Blue) which was analysed in both malignant (D, E, F) and non-malignant (A, B, C) prostate tissue. Whole tissue cores (A and D) were imaged on an Olympus IX81 confocal system at low magnification. Intensity of fluorescent signals were optimised at the beginning of each study to prevent oversaturation for each fluorophore. Quantification of co-localization was carried out on images gathered from a randomly selected area of the tissue core using a 40x objective zoom (E is malignant and B is non-malignant) on an Olympus IX81 confocal system. The image was further magnified using a 6x digital zoom. Each image was then deconvolved (C and F) and the GIC was calculated using Huygens software (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0186047#sec003" target="_blank">Materials and methods</a>). Scale bar = 10 μm.</p

ROC of RPS19, RPS21, and RPS24 in prostate tissue.

<p>ROC curve to demonstrate the potential ability of a label to differentiate between malignant and non-malignant prostate tissue. The probit function was used for area fraction per amount of tissue for ROC analysis of RPS19, RPS21, and RPS24. The dotted line is representative of an area under the curve of 0.5 which shows no distinction between the two classifiers (e.g. normal vs malignant).</p

Box plots of co-localization of RPS proteins.

<p>Global intersection coefficient (GIC) was calculated using deconvolved, high magnification, fluorescent images (represented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0186047#pone.0186047.g005" target="_blank">Fig 5</a>) using Huygens software (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0186047#sec003" target="_blank">Materials and methods</a>). Box plots of GIC were created for comparison between malignant (red bordered bars) and non-malignant (green bordered bars) co-localization for RPS21/RPS19 (green/red bars), RPS21/RPS24 (green/blue bars), and RPS19/RPS24 (red/blue bars). Significance of the difference in GIC values for malignant vs non-malignant prostate tissue was calculated using the Mann Whitney U test (* = <i>p</i><0.01).</p

ROC of putative prostate cancer biomarkers.

<p>ROC curve demonstrates the discriminating performance of the protein expression in the differentiation between malignant and non-malignant tissue cores using the area fraction (probit) data for BTF3, HINT1, NDRG1 and ODC1. The operating characteristic values are given in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0084295#pone.0084295.s005" target="_blank">Table S1</a>. The solid line represents the ROC area of 0.5. </p

Multi-labeled immunofluorescence for BTF3, HINT1 and NDRG1.

<p>Co-expression of three proteins BTF3 (FITC-green), HINT1 (Cy3- blue) and NDRG1 (Cy5-red) in non-malignant (A) and malignant prostate tissue cores (B); images are representative tissue cores from the tissue array with over 200 samples. The three antibodies were incubated simultaneously and labeled with three Cy5 –red different fluorophores using Bond automated staining system. Whole tissue cores were imaged using a Leica confocal system and 4x4 tiling at exactly the same settings for comparison. Images were zoomed 3x using digital zoom in ImageJ. Scale bar =100µm.</p

Expression analysis of Ox-DAB signal in prostate tissue.

<p>Area fraction (AF) was calculated by dividing the amount of signal, in pixels, by the total number of pixels representing tissue per an unbiased signal quantification protocol (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0186047#sec003" target="_blank">Materials and methods</a>- Signal quantification). Each bar is representative of the AF of a single TA core. Malignant cores are represented by red and non-malignant cores are represented by green. A significant difference between AUC values was seen for each protein that was tested (p<0.0001).</p

Changes in RPS protein expression with Gleason grade.

<p>Box plots were created in Origin software to show the changes in expression of RPS19, RPS21, and RPS24, in different Gleason grades. Brackets represent significant differences in protein expression (p<0.05). A significant change was observed between RPS19 Gleason grades 5 and 8, and 6 and 8 (p = 0.007 and p = 0.028 respectively). A significant difference was also observed in RPS21 Gleason grades 5 and 8 (p = 0.0488).</p