List of p values of DNA repair proteins in univariate analysis of the correlation with overall survival.

<p>List of p values of DNA repair proteins in univariate analysis of the correlation with overall survival.</p

Association of XPF scoring by pathologist scores versus machine assisted image analysis and quantitation.

<p>Comparisons are made between alternative scoring strategies for immunohistochemistry with the XPF for each head and neck cancer patient. Machine assisted scoring for XPF was determined based on percentage of nuclei with 1+ (weak), 2+ (medium), 3+ (high) intensity Pathologist scores were Intensity (I). Correlation plots as shown are computed for similarity with an R-value of 0.79.</p

Immunohistochemistry (IHC) staining pattern of the DNA repair biomarkers.

<p>A. The FFPE whole sections from 37 HNSCC patient samples were stained by IHC using the antibodies against DNA repair biomarkers (XPF, ERCC1, FANCD2, MLH1, pMK2, PAR, PARP1) according to the protocol described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0102112#s2" target="_blank"><i>Materials and Methods</i></a>. The stained tissue on the slide was scanned into a digital pathology platform (Aperio) and images were viewed digitally, magnification 10X. As noted, subcellular localization of pMK2 is in either Nuclear (N), or Nuclear (N) + Cytoplasmic (C), staining patterns of pMK2 in these cancer tissues is shown as indicated, magnification 20X. Nuclear foci in head neck cancer cells were shown for γH2AX and FANCD2 in the lower panel as indicated, magnification 40X. B. Examples of varying biomarker expression in head and neck cancer tissue specimens stained with XPF, FANCD2, MLH1 are shown. Patient distribution of XPF, FANCD2, MLH1 scores are plotted. C. Differences in the staining intensity and distribution of XPF (NER), MLH1 (MMR), PAR (BER), FANCD2 (FA/HR), pMK2 and γH2AX (DDR) in parabasal (pb) and nonparabasal (non-pb) layer cells from specimens of one representative HNSCC patient were shown as indicated.</p

Univariate analysis of XPF biomarker scores shows improved response prediction to induction chemotherapy in head and neck cancer.

<p>The chart shows that univariate analysis of the XPF biomarker scores relative to the discrimination between Responder subgroups. The primary outcome measurement was response to induction chemotherapy.</p

Correlation of expression levels of pMK2 with overall survival.

<p>Overall survival estimated by best response to induction chemotherapy using Kaplan-Meier survival curves based on the nuclear staining intensity and quantitation of pMK2 determined by pathologists' scores as NQ (Nuclear Quantity).</p

The ExPeCT (Examining Exercise, Prostate Cancer and Circulating Tumour Cells) trial: study protocol for a randomised controlled trial

Background: Prostate cancer (PrCa) is the second most common cancer in Ireland. Many men present with locally advanced or metastatic cancer for whom curative surgery is inappropriate. Advanced cancer patients are encouraged to remain physically active and therefore there is a need to investigate how patients with metastatic disease tolerate physical activity programmes. Physical activity reduces levels of systemic inflammatory mediators and so an aerobic exercise intervention may represent an accessible and cost-effective means of ameliorating the pro-inflammatory effects of obesity and subsequently decrease poor cancer-specific outcomes in this patient population. This study will assess the feasibility and safety of introducing a structured aerobic exercise intervention to an advanced cancer population. This study will also examine if the evasion of immune editing by circulating tumour cells (CTCs) is an exercise-modifiable mechanism in obese men with prostate cancer. Methods: This international multicentre prospective study will recruit men with metastatic prostate cancer. Participants will be recruited from centres in Dublin (Ireland) and London (UK). Participants will be divided into exposed and non-exposed groups based on body mass index (BMI) ≥ 25 kg/m2 and randomised to intervention and control groups. The exercise group will undertake a regular supervised aerobic exercise programme, whereas the control group will not. Exercise intensity will be prescribed based on a target heart rate monitored by a polar heart rate monitor. Blood samples will be taken at recruitment and at 3 and 6 months to examine the primary endpoint of platelet cloaking of CTCs. Participants will complete a detailed questionnaire to assess quality of life (QoL) and other parameters at each visit. Discussion: The overall aim of the ExPeCT trial is to examine the relationship between PrCa, exercise, obesity, and systemic inflammation, and to improve the overall QoL in men with advanced disease. Results will inform future work in this area examining biological markers of prognosis in advanced prostate cancer. Trial registration: Clinicaltrials.gov NLM identifier: NCT02453139 . Registered on 12 May 2015. This document contains excerpts from the ExPeCT trial protocol Version 1.5, 28 July 2016.</p

Heterogeneous expression of monoclonal anti-TLR4.

<p>A: variable staining observed in adjacent epithelium within a benign serous cystadenoma (20x). B: focal strong staining within a serous carcinoma (40x).</p

MyD88 expression and survival (Kaplan-Meier curves: median survival time shown in months).

<p>MyD88 positive tumours (n = 12) had significantly reduced progression-free survival (A) and overall survival (B) (p = 0.018 and p = 0.008, respectively).</p

The effect of silencing MyD88 and TLR4 mRNA on the chemoresponsive properties of SKOV-3 cells.

<p>SKOV-3 cells were left untransfected (Unt), transfected with negative control siRNA (siNeg), MyD88 targeting siRNA (siMyD88) or TLR4 targeting siRNA (siTLR4). The transfected cells were incubated for 72 hrs before either harvesting for mRNA analysis (A), for protein analysis (B) or treatment with paclitaxel (C). (A) MyD88 and TLR4 mRNA expression levels were evaluated by TaqMan RT-PCR. MyD88 and TLR4 mRNA expression was normalised to that of an endogenous control, B2M, and calibrated to that of untreated cells to establish the relative percentage of mRNA expression (n = 3, mean +SD). (B) MyD88 and TLR4 mRNA expression levels were evaluated by western blot analysis. GAPDH was used as a loading control. (C) Transfected cells were either left untreated, treated with DMSO (vehicle control) or 3.5 nM of paclitaxel (IC25). 48 hrs post treatment, cell viability was assessed by means of the CCK-8 assay. % cell viability rate was calculated by comparing the absorbance values for the vehicle control to the corresponding paclitaxel treated samples. Results are expressed as mean +SD, n = 3; *p<0.05, **p<0.01 (un-paired Student's t-test).</p

TLR4/MyD88 and overall survival (Kaplan-Meier curves; median survival shown in months).

<p>Survival was longer in MyD88 (B) negative cases (by 19 months; p<0.05). The difference in survival associated with TLR4 (A) is not significant (p>0.5).</p