Definition of treatment response on the IPSS, BII and PGI-I after 12 weeks treatment with tadalafil or placebo as used in the clinical data mining analysis.

<p>BII, BPH Impact Index; IPSS, International Prostate Symptom Score; PGI-I, Patient Global Impression of Improvement; QoL, quality of life.</p><p>Definition of treatment response on the IPSS, BII and PGI-I after 12 weeks treatment with tadalafil or placebo as used in the clinical data mining analysis.</p

Predictors of Individual Response to Placebo or Tadalafil 5mg among Men with Lower Urinary Tract Symptoms Secondary to Benign Prostatic Hyperplasia: An Integrated Clinical Data Mining Analysis

<div><p>Background</p><p>A significant percentage of patients with lower urinary tract symptoms (LUTS) secondary to benign prostatic hyperplasia (BPH) achieve clinically meaningful improvement when receiving placebo or tadalafil 5mg once daily. However, individual patient characteristics associated with treatment response are unknown.</p><p>Methods</p><p>This integrated clinical data mining analysis was designed to identify factors associated with a clinically meaningful response to placebo or tadalafil 5mg once daily in an individual patient with LUTS-BPH. Analyses were performed on pooled data from four randomized, placebo-controlled, double-blind, clinical studies, including about 1,500 patients, from which 107 baseline characteristics were selected and 8 response criteria. The split set evaluation method (1,000 repeats) was used to estimate prediction accuracy, with the database randomly split into training and test subsets. Logistic Regression (LR), Decision Tree (DT), Support Vector Machine (SVM) and Random Forest (RF) models were then generated on the training subset and used to predict response in the test subset. Prediction models were generated for placebo and tadalafil 5mg once daily Receiver Operating Curve (ROC) analysis was used to select optimal prediction models lying on the ROC surface.</p><p>Findings</p><p>International Prostate Symptom Score (IPSS) baseline group (mild/moderate vs. severe) for active treatment and placebo achieved the highest combined sensitivity and specificity of 70% and ~50% for all analyses, respectively. This was below the sensitivity and specificity threshold of 80% that would enable reliable allocation of an individual patient to either the responder or non-responder group</p><p>Conclusions</p><p>This extensive clinical data mining study in LUTS-BPH did not identify baseline clinical or demographic characteristics that were sufficiently predictive of an individual patient response to placebo or once daily tadalafil 5mg. However, the study reaffirms the efficacy of tadalalfil 5mg once daily in the treatment of LUTS-BPH in the majority of patients and the importance of evaluating individual patient need in selecting the most appropriate treatment.</p></div

Secondary Results (Placebo).

<p>*Model on ROC surface with best performance if false positive and false negative errors are equally important.</p><p>** Alpha-Blockers, beta-blockers, calcium channels blockers, angiotensin converting enzyme inhibitors, angiotensin receptor blockers and diuretics.</p><p>BII, BPH Impact Index; CI, confidence interval; ED, erectile dysfunction; IIEF, International Index of Erectile Function; IPSS, International Prostate Symptom Score; N, no; PGI-I, Patient Global Impression of Improvement; PGISS, Patient Global Incontinence Severity Score; QoL, quality of life; ROC, Receiver Operating Curve; SHBG, sex hormone binding globulin; Y, yes.</p><p>Secondary Results (Placebo).</p

Exploratory Results.

<p>Models were generated on dataset excluding testosterone, alcohol frequency, Qmax, SHBG, Albumin, PGI, and PSA</p><p>BII, BPH Impact Index; CI, confidence interval; DT, Decision Tree; IPSS, International Prostate Symptom Score; MCID, Minimally Clinically Important Differences; PGI-I, Patient Global Impression of Improvement; PSA, prostate specific antigen; Q_max, maximal flow rate; QoL, quality of life; RF, Random Forest; SHBG, sex hormone binding globulin.</p><p>Exploratory Results.</p

Primary results for both treatment groups.

<p>*Model on ROC surface with best performance if false positive and false negative errors are equally important.</p><p>CI, confidence interval; ED, erectile dysfunction; IPSS, International Prostate Symptom Score; MCID, Minimal Clinically Important Differences; N, no; PSA, prostate specific antigen; ROC, Receiver Operating Curve; Y, yes.</p><p>Primary results for both treatment groups.</p

Secondary Results (Tadalafil 5mg once daily).

<p>*Model on ROC surface with best performance if false positive and false negative errors are equally important.</p><p>** Alpha-Blockers, beta-blockers, calcium channels blockers, angiotensin converting enzyme inhibitors, angiotensin receptor blockers and diuretics.</p><p>BII, BPH Impact Index; CI, confidence interval; IIEF, International Index of Erectile Function; IPSS, International Prostate Symptom Score; N, no; PGI, Patient Global Impression of Improvement; Q_max, maximal flow rate; QoL, quality of life; ROC, Receiver Operating Curve; Y, yes.</p><p>Secondary Results (Tadalafil 5mg once daily).</p

Additional file 3: of Tadalafil 5 mg once daily for the treatment of erectile dysfunction during a 6-month observational study (EDATE): impact of patient characteristics and comorbidities

Kaplan-Meier estimation in different age groups for time to discontinuation of tadalafil OaD treatment. (PDF 109 kb

Workflow of the DPI-ELISA screen.

<p>(A) Design of the double stranded (ds) DNA probes. Each dsDNA library probe contains a variable library region of up to 20 base pairs, which covers up to 30 different hexanucleotide sequences. Each of the probes is flanked by T/A repeats as linkers. As an example, the double-stranded DNA-sequence of the variable region in dsDNA library probe 18 is shown (right). A total of 30 overlapping, non-redundant hexanucleotides are distributed on both DNA-strands. (B) The individually synthesized DNA probes are annealed and 2 pmol of each dsDNA probe are immobilized in a single well. Hence, each individual dsDNA probe is assigned a specific plate position. Distribution of dsDNA probes on the 384 well plate according to the scheme by using robotics. (C) The DNA coated plate is probed and analyzed according to the DPI-ELISA protocol <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075177#pone.0075177-Brand1" target="_blank">[46]</a> by using robotics. (D) The photometric readout is normalized to the mean and analyzed to identify positively bound DNA motifs from which a DNA binding consensus is derived.</p

Proof-of-principle experiment using the WRKY11 DNA-binding domain (DBD).

<p>(A) Two individual DPI-ELISA screens were performed with WRKY11 DBD; a plate scan of replicate #2 is shown. (B) All 384 probes of the plate replicate #2 were hierarchically ranked according to their relative normalized absorbance values. : positive and negative controls, : dsDNA library probes, ???: indicate positive probes. Background shading indicates confidence interval (p = 0.05) (C) Comparison of the two biological replicate DPI-ELISA screens with WRKY11 DBD. Relative normalized absorbance values of replicate #1 (x-axis) and replicate #2 (y-axis) were plotted against each other to demonstrate the high reproducibility (left). : positive and negative controls, : dsDNA library probe, : indicate positive probes. <b>- - -</b>: borders of significance (p≤0.05). Probe name, plate position and rank of positively bound dsDNA library probes that clustered with the positive control probes are given for replicates #1 and #2 (right). (D) The positive dsDNA probe sequences were aligned according to DREME <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0075177#pone.0075177-Bailey1" target="_blank">[54]</a>. Pos.: plate position. Finally, the DNA-binding consensus was derived (E).</p

Identification of a DNA-binding motif for <i>At</i>TIFY1.

<p>(A) All 384 probes of the <i>At</i>TIFY1 DPI-ELISA screen were hierarchically ranked according to their relative normalized absorbance values. : positive and negative controls, : dsDNA library probes, : indicate positive probes. Background shading indicates confidence interval (p = 0.05). Alignments of positive dsDNA probes were used to deduce the binding consensus next to the graph. (B) Sequence of the significantly bound dsDNA probe 38 that was chosen for further studies of <i>At</i>TIFY1 - DNA-interaction. (C) Analysis of the <i>At</i>TIFY1-binding motif by sequential mutagenesis of the dsDNA probe 38 by quantitative DPI-ELISA. Mutagenesis of the identified <i>At</i>TIFY1 DNA-binding motif results in a altered relative signal intensity of <i>At</i>TIFY1 to the DNA in DPI-ELISA experiments. Relative binding values [%] for the mutated probes are given in comparison to the non- mutagenized dsDNA probe 38. Error bars represent the absolute error of two technical replicates. Significantly bound probes (p<0.05) are indicated by asterisks and are used for the binding consensus that is given next to the graph.</p