Discrimination of delays of reinforcement in aversive conditioning.

<p>(a) The protein level changes of CA125 and LRG1 were confirmed in the larger sample set 2. (b) Marker distributions for the EOC patients with different histological subtypes.</p

Development of a quantitative western blot assay for DR6.

<p><b>A</b>. Representative quantitative immunofluorescent western blot analysis of DR6 serum protein expression from ovarian cancer patients and donor control patients. <b>B</b>. Concordance of western blot analysis of replicate samples in independent experiments (r = 0.97). <b>C</b>. Concordance of inter-day (gray) and intra-day (black) replicates of reference serum standards. <b>D</b>. Serum protein levels from control and patients with the indicated cancer types. Only adult sarcoma patients demonstrated a statistically significant increased expression of serum DR6 protein compared to control.</p

Validation of LRG1 as a Potential Biomarker for Detection of Epithelial Ovarian Cancer by a Blinded Study

<div><p>Background</p><p>Leucine-rich alpha-2-glycoprotein (LRG1) was found to be differentially expressed in sera from patients with Epithelial Ovarian Cancer (EOC). The aim of this study is to investigate the performance of LRG1 for detection of EOC, including early stage EOC, and to evaluate if LRG1 can complement CA125 in order to improve EOC detection using two independent blinded sample sets.</p><p>Methods and Results</p><p>Serum LRG1 and CA125 were measured by immunoassays. All assays were performed blinded to clinical data. Using the two independent sample sets (156 participants for sample set 1, and 233 for sample set 2), LRG1 was differentially expressed in EOC cases as compared to healthy, surgical, and benign controls, and its performance was not affected by the conditions of blood collection. The areas under the ROC curve (AUC) for LRG1 in differentiating EOC cases from non-cases were 0.797 and 0.786 for sample set 1 and 2. For differentiating EOC cases from healthy controls, the AUC values for LRG1 were 0.792 and 0.794. At a fixed specificity of 95%, LRG1 detects 52%, and 53.5% of EOC cases from healthy controls for sample set 1 and 2. When combining LRG1 and CA125, the AUC value increased to 0.927, which was improved compared to CA125 (AUC=0.916) (<i>p</i>=0.008) alone in distinguishing EOC cases from non-cases. More importantly, LRG1 also showed potential performance in differentiating early stage EOC from non-cases with an AUC of 0.715 for sample set 1, and 0.690 for sample set 2. The combination of LRG1 and CA125 resulted in an AUC of 0.838, which outperforms CA125 (AUC=0.785) (<i>p</i>=0.018) in detecting early stage EOC cases from non-cases using the larger sample set.</p><p>Conclusions</p><p>LRG1 could be a useful biomarker alone or in combination with CA125 for the diagnosis of ovarian cancer.</p></div

DR6 mRNA expression.

<p>Expression of DR6 mRNA in cancer expression arrays. A. DR6 mRNA expression in microarrays of the indicated tumor types determined using Oncomine™ software. Thick lines indicate the median, thin lines indicate the 90^th/10^th percentiles, box indicates 25^th–75^th percentiles, dots indicate the minimum and the maximum. B. Quantitative PCR confirming DR6 mRNA expression levels in the indicated tumor types. Average expression with standard error are indicated (n = 2–5 tumors in each group).</p

Sarcoma patients have significant elevation of DR6 serum protein.

<p><b>A.</b> DR6 protein expression in a panel of 39 healthy controls and 71 patients with adult sarcoma. <b>B</b> DR6 levels in healthy controls and in sarcoma patients separated into chemo-naïve and chemo-refractory sub-groups. <b>C</b>. Receiver operator characteristic (ROC) curve for DR6 serum level in sarcoma patients versus controls. Area under the curve is 82.1%. <b>D</b>. Sarcoma patient DR6 serum protein levels based on histologic subtype. Averages are indicated by a long horizontal line. Brackets indicate standard deviations.</p

Characteristics of Sarcoma Patients Evaluated.

<p>NA-not applicable.</p

Correlating change in DR6 serum protein over time with response to therapy.

<p><b>A</b>. Waterfall plot of change in DR6 serum protein values prior to and after therapy with sirolimus and cyclophosphamide in patients with advanced sarcoma; patients with stable disease are indicated by black bars and patients with progressive disease are indicated in Gray. <b>B</b>. Sensitivity, specificity, negative predictive value (NPV) and positive predictive value (PPV) for the change in DR6 serum protein levels to predict stable disease versus disease progression in sarcoma patients on therapy. <b>C</b>. Waterfall plot of change in DR6 serum protein values prior to and after therapy with sirolimus and cyclophosphamide in patients with (1) leiomyosarcoma and (2) liposarcoma. Patients with stable disease are indicated by black bars and patients with progressive disease are indicated in Gray.</p

ROC curves comparing marker concentrations in cases to healthy controls for sample set 1.

<p>(a) ROC analyses for CA125 and LRG1 to differentiate EOC from healthy controls. (b) ROC analyses for CA125 and LRG1 to differentiate early stage EOC from healthy controls.</p

ROC curves comparing marker concentrations in cases to non-cases for sample set 1.

<p>(a) ROC analyses for CA125 and LRG1 to differentiate EOC from non-cases. (b) ROC analyses for CA125 and LRG1 to differentiate early stage EOC from non-cases.</p

Multimarker panel analysis.

<p>The performance of multimarker panels in distinguishing EOC/ early stage EOC from non-cases using sample set 1and sample set 2.</p