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Lindsay A Brown1,2, Jeremy Hoog3, Suet-Feung Chin4,5, Yu Tao3, Abd Alnaser Zayed1,2, Koei Chin6, Andrew E Teschendorff4,5, John F Quackenbush7, John C Marioni8, Samuel Leung9,10, Charles M Perou11, Torsten O Neilsen9,10, Matthew Ellis3, Joe W Gray6, Philip S Bernard7, David G Huntsman1,2,9,10,12, and Carlos Caldas4,5,12 1Center for Translational and Applied Genomics, and the British Columbia Cancer Agency, Vancouver, British Columbia V5Z 4E6, Canada.

Purpose: Basal-like breast cancer is associated with high grade, poor prognosis, and younger patient age. Clinically, a triple-negative phenotype definition [estrogen receptor, progesterone receptor, and human epidermal growth factor receptor (HER)-2, all negative] is commonly used to identify such cases. EGFR and cytokeratin 5/6 are readily available positive markers of basal-like breast cancer applicable to standard pathology specimens. This study directly compares the prognostic significance between three- and five-biomarker surrogate panels to define intrinsic breast cancer subtypes, using a large clinically annotated series of breast tumors. Experimental Design: Four thousand forty-six invasive breast cancers were assembled into tissue microarrays. All had staging, pathology, treatment, and outcome information; median follow-up was 12.5 years. Cox regression analyses and likelihood ratio tests compared the prognostic significance for breast cancer death-specific survival (BCSS) of the two immunohistochemical panels. Results: Among 3,744 interpretable cases, 17% were basal using the triple-negative definition (10-year BCSS, 6 7%) and 9% were basal using the five-marker method (10-year BCSS, 62%). Likelihood ratio tests of multivariable Cox models including standard clinical variables show that the five-marker panel is significantly more prognostic than the three-marker panel. The poor prognosis of triple-negative phenotype is conferred almost entirely by those tumors positive for basal markers. Among triple-negative patients treated with adjuvant anthracycline-based chemotherapy, the additional positive basal markers identified a cohort of patients with significantly worse outcome. Conclusions: The expanded surrogate immunopanel of estrogen receptor, progesterone receptor, human HER-2, EGFR, and cytokeratin 5/6 provides a more specific definition of basal-like breast cancer that better predicts breast cancer survival.

Background HER2 gene amplification and/or protein overexpression in breast cancer is associated with a poor prognosis and predicts response to anti-HER2 therapy. We examine the natural history of breast cancers in relationship to increased HER2 copy numbers in a large population-based study. Patients and Methods HER2 status was measured by fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) in approximately 1,400 breast cancer cases with greater than 15 years of follow-up. Protein expression was evaluated with two different commercially-available antibodies. Results We looked for subgroups of breast cancer with different clinical outcomes, based on HER2 FISH amplification ratio. The current HER2 ratio cut point for classifying HER2 positive and negative cases is 2.2. However, we found an increased risk of disease-specific death associated with FISH ratios of >1.5. An ‘intermediate’ group of cases with HER2 ratios between 1.5 and 2.2 was found to have a significantly better outcome than the conventional ‘amplified’ group (HER2 ratio >2.2) but a significantly worse outcome than groups with FISH ratios less than 1.5. Conclusion Breast cancers with increased HER2 copy numbers (low level HER2 amplification), below the currently accepted positive threshold ratio of 2.2, showed a distinct, intermediate outcome when compared to HER2 unamplified tumors and tumors with HER2 ratios greater than 2.2. These findings suggest that a new cut point to determine HER2 positivity, at a ratio of 1.5 (well below the current recommended cut point of 2.2), should be evaluated.

Gastrointestinal stromal tumors (GISTs) are mesenchymal neoplasms found in the gastrointestinal tract. The purpose of this study was to evaluate whether morphometric measurements could complement tumor size and mitotic activity in risk evaluation. Nuclear roundness and ellipse axis ratio were found to correlate with tumor size, mitotic activity, nuclear atypia, and hemorrhage. Morphometric variables in 422 GISTs were significant for overall survival in univariate analyses but did not retain independent significance in multivariate analyses incorporating mitotic count and tumor size. Traditional variables, together with sex, location of primary tumor, and nuclear atypia, seem to be the best parameters for prognostic evaluation. (J Histochem Cytochem 56:139–145, 2008)

There is currently great interest in reuse of digital learning resources, from single items to multi-task activities, to whole units or programmes of study. Associated with this interest is the ongoing development of tools to enable such resources to be placed in or recovered from repositories, to facilitate searching for them and their instantiation on different learning platforms. Many of these tools are highly technical, in terms of both technology and terminology, and they are the province of learning technologists and computer scientists, rather than teachers. Concentration on such tools begs the question as to whether teachers and learners will truly benefit from reuse of learning resources. This paper describes the re-purposing of a learning activity created in a university in the US for use in two UK universities. The case study approach facilitates comparison of the technical and content issues that were addressed and resolved. The authors formulate and explore a proposition about the validity of an ‘object based’ approach to learning resources and conclude with a plea for better tools for teachers., JRC.H.3-Global environement monitoring

RESULTS: Sixty-five hysterectomy specimens were examined for depth of thermal spread. There was no difference in mean depth of thermal spread at the anterior (1.11 mm vs 1.05 mm; p1⁄40.6) or posterior margin (1.04 mm vs 0.91 mm; p1⁄40.2) between standard energy and V mode , respectively. There was no difference in incidence of vaginal cuff dehiscence (p1⁄40.31), with one dehiscence in the standard group. Rate of vaginal cuff granulation tissue, dehiscence or infection was 24% overall, with no difference between groups.

562 Background: We previously showed that baseline patient/tumour characteristics, or prior treatment affected cause of death in NCIC CTG endocrine therapy trials (MA.14, MA.17, MA.27), in particular older age was associated with non-breast cancer death. We now examine whether PAM50 intrinsic subtypes were differentially associated with cause of death in a cohort of British Columbia tamoxifen-treated patients. Methods: Patients were 718 post-menopausal women, treated with tamoxifen, without adjuvant chemotherapy who had tumours which were Luminal A, Luminal B, Basal, or HER2 intrinsic subtype and known cause of death [cancer-specific (BrCa); other cause (OC)]. We tested whether intrinsic subtype and other patient and tumour characteristics were associated with 1) all cause mortality, and if so, 2) cause-specific mortality. We also fit step-wise forward cause-specific adjusted models. Results: Patients were followed a median of 11.7 years. In women

Rationale: FBxW7 is a tumor suppressor gene that has been shown to be linked to ubiquitin-dependent proteolysis and regulation of the G1-S cell cycle checkpoint. FBxW7 mutations have been implicated in the impaired degradation of NOTCH, JUN, and cMYC, all of which have been demonstrated as key players in the ovarian cancer pathobiology. In this study we characterized the functional and prognostic significance of FBxW7 in ovarian cancer. Methods: Platinum-sensitive (A2780) and resistant (C200 and SKOV3) ovarian cancer cell lines were utilized, as well as cell lines derived from normal ovarian epithelium (IOSE). Ovarian cancer cell lines were transfected with FBxW7 constructs using Lipofectamine 2000tm. qPCR was performed to determine levels of gene and microRNA expression. Western blot analysis was performed to assess protein levels. Cell proliferation assays and FACS were performed determine cell proliferation and viability. A tissue microarray was used for immunohistochemical staining using FBxW7 antibody. Tissue stains were scored manually. Staining intensities were correlated with patient outcomes to calculate the overall survival and progression-free survival using Kaplan Meier methods and log-rank tests. Results: Upregulation of FBxW7 in platinum-sensitive cell lines decreases the substrates of FBxW7 ubiquitination, such as cMYC, NOTCH, and JUN. However, similar levels of FBxW7 expression did not decrease these substrate levels in platinum-resistant cell lines. microRNAs associated with stemness and chemoresistance also had differential expression profiles with FBxW7 transient transfection. Cell proliferation assays also highlighted differences between these sensitive and resistant cell lines. For tissue array analysis, 516 samples were used for FBxW7 IHC staining where 206 (40.2%) of samples were with high-grade serous histology. The majority of samples had FBxW7 staining of high intensity (273; 52.9%). There was no relationship between overall survival or progression-free survival and FBxW7 tissue staining in the total group. However, when analyzing only the high-grade serous histology, there was a trend of increased expression of FBxW7 being associated with better progression-free survival (p=0.051). Conclusions: Our studies suggest that FBxW7 have a dual role in ovarian cancer, having less effect on those cell lines associated with chemoresistance and stemness. In ovarian tumor tissues, there was a trend for elevated levels of FBxW7 expression and better progression free survival. Further studies on FBxW7 and its role in ovarian cancer pathogenesis are warranted. Citation Format: Elizabeth Louise Dickson, Lihua Li, Samuel Leung, Christine Chow, Rachel Isaksson Vogel, David Huntsman, C. Blake Gilks, Subbaya Subramanian. FBxW7 duality in ovarian cancer: Novel insight into ovarian cancer pathogenesis. [abstract]. In: Proceedings of the AACR Special Conference on Advances in Ovarian Cancer Research: From Concept to Clinic; Sep 18-21, 2013; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2013;19(19 Suppl):Abstract nr A17.

Purpose: To describe early outcome and technical treatment aspects of four patients with pediatric genitourinary malignancy receiving local therapy using interstitial brachytherapy at the Peter MacCallum Cancer Institute since 1991. Methods and Materials: Two patients with primary bladder tumors and two patients with primary vaginal tumors received interstitial brachy-therapy with needle-template techniques following chemotherapy. Two patients were treated for salvage following local failure after primary chemotherapy and two patients electively after initial induction chemotherapy, one following limited surgical excision. Three patients were treated exclusively with brachytherapy with doses of 50-55 Gy (LDR), and one patient also received external beam radiation. Results: Resultant implants, dosimetry, and dose prescriptions are described and illustrated. Follow-up is short (14-27 months), but all patients remain well without significant sequelae. One child required salvage surgery for local failure. Conclusion: Interstitial brachytherapy using needle-template techniques described is an ideal method of local ablative therapy for pediatric genitourinary malignancy. Functional and late sequelae should be less than that of radical surgery or external beam radiation. There are technical and practical advantages compared with other brachytherapy techniques described in the literature

Background: Breast cancer is heterogeneous and the existing prognostic classifiers are limited in accuracy, leading to unnecessary treatment of numerous women. B-cell lymphoma 2 (BCL2), an antiapoptotic protein, has been proposed as a prognostic marker, but this effect is considered to relate to oestrogen receptor (ER) status. This study aimed to test the clinical validity of BCL2 as an independent prognostic marker. Methods: Five studies of 11 212 women with early-stage breast cancer were analysed. Individual patient data included tumour size, grade, lymph node status, endocrine therapy, chemotherapy and mortality. BCL2, ER, progesterone receptor (PR) and human epidermal growth factor receptor 2 (HER2) levels were determined in all tumours. A Cox model incorporating the time-dependent effects of each variable was used to explore the prognostic significance of BCL2. Results: In univariate analysis, ER, PR and BCL2 positivity was associated with improved survival and HER2 positivity with inferior survival. For ER and PR this effect was time dependent, whereas for BCL2 and HER2 the effect persisted over time. In multivariate analysis, BCL2 positivity retained independent prognostic significance (hazard ratio (HR) 0.76, 95% confidence interval (CI) 0.66–0.88, P

Metastasis of primary malignancies is a multi-step process and remains the principal cause of cancer deaths. Carbonic anhydrase IX (CAIX) is a hypoxia inducible protein and a poor prognostic marker for several types of cancer, including breast cancer. However, the functional role of CAIX in the metastatic progression of breast cancer is unclear. Here, we have investigated its role in the growth and metastasis of breast tumors. Orthotopic mouse mammary tumors derived from metastatic 4T1 and 66cl4 cells or non-metastatic 67NR cells were examined for levels of proliferation (BrdU), hypoxia, (pimonidazole), perfusion (DiOC7), vasculature (CD31), apoptosis (TUNEL) and lymphangiogenesis (LYVE-1). Metastatic 4T1 and 66cl4 tumors expressed a hypoxia gene signature and were characterized as being poorly vascularized, with high levels of hypoxia. Large numbers of apoptotic cells and well developed intratumoral lymphatic vessels were also evident. Inhibition of expression of CAIX in the metastatic 4T1 cells by stable expression of short hairpin RNA (shRNA) resulted in cell death and reversal of extracellular acidosis in hypoxia in vitro, dramatic regression of tumors in vivo, and inhibition of metastasis. These properties were rescued by constitutive expression of human CAIX. Treatment of mice harboring 4T1 tumors with a novel CAIX-specific inhibitor resulted in significant inhibition of tumor growth. Interrogation by immunohistochemistry of a large (3992 patient samples) primary breast tumor tissue microarray showed that CAIX expression was significantly associated with worse distant relapse free survival (p This work was supported by the Canadian Breast Cancer Research Alliance, with special funding from the Canadian Breast Cancer Foundation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 455.

Introduction Primary breast cancer involving four or more axillary lymph nodes carries a poor prognosis. We hypothesized that use of an immunohistochemical biomarker scoring system could allow for identification of variable risk subgroups. Methods Patients with four or more positive axillary nodes were identified from a clinically annotated tissue microarray of formalin-fixed paraffin-embedded primary breast cancers and randomized into a 'test set' and a 'validation set'. A prospectively defined prognostic scoring model was developed in the test set and was further assessed in the validation set combining expression for eight biomarkers by immunohistochemistry, including estrogen receptor, human epidermal growth factor receptors 1 and 2, carbonic anhydrase IX, cytokeratin 5/6, progesterone receptor, p53 and Ki-67. Survival outcomes were analyzed by the Kaplan–Meier method, log rank tests and Cox proportional-hazards models. Results A total of 313 eligible patients were identified in the test set for whom 10-year relapse-free survival was 38.3% (SEM 2.9%), with complete immunohistochemical data available for 227. Tumor size, percentage of positive axillary nodes and expression status for the progesterone receptor, Ki-67 and carbonic anhydrase IX demonstrated independent prognostic significance with respect to relapse-free survival. Our combined biomarker scoring system defined three subgroups in the test set with mean 10-year relapse-free survivals of 75.4% (SEM 7.0%), 35.3% (SEM 4.1%) and 19.3% (SEM 7.0%). In the validation set, differences in relapse-free survival for these subgroups remained statistically significant but less marked. Conclusion Biomarkers assessed here carry independent prognostic value for breast cancer with four or more positive axillary nodes and identified clinically relevant prognostic subgroups. This approach requires refinement and validation of methodology.

7211 Background: The purpose of this study is to use a large patient population to identify immunohistochemical (IHC) biomarkers to enable improved prognostication in patients with non-small cell lung cancer (NSCLC). Methods: A tissue microarray was constructed using duplicate 0.6 mm cores of formalin-fixed paraffin embedded tissue blocks from 609 patients with NSCLC. IHC was used to detect 11 biomarkers including EGFR, HER2, HER3, p53, p63, Bcl-1, Bcl-2, TTF-1, CEA, Ch, and SNP. A clinical database was created prospectively at the time of tissue collection. Survival outcomes were obtained from a Provincial Cancer Registry database. Results: Male to female ratio was 400:209; median age 63yrs (range 35–82); median survival 3.5yrs (mean 5.7). All specimens were reviewed: 243 adenocarcinoma (ACA), 272 squamous cell carcinoma (SCC), 35 large cell carcinoma, 32 non-small cell carcinoma NOS, and 6 other (giant cell carcinoma). 21 patients with other histologies were excluded. Survival data for 535 cases was available. As of June 2005, 429 patients (80%) had died; of these 286 (54%) died of lung cancer, 117 (22%) died of other known causes, and for 26 (5%) the cause of death was not available. Bcl-2 (p = 0.007) was the only biomarker to predict better overall survival (OS). Bcl-2 (p = 0.021) and p63 (p = 0.025) were significant for disease specific survival (DSS) in all NSCLC. Analysis of the subgroups indicated that p63 was significant (p = 0.039) for DSS in squamous cell carcinoma (SCC) but not for adenocarcinoma (ACA) (p = 0.81). Bcl-2 was not significant for DSS in either subgroup (p = 0.28 for SCC, p = 0.112 for ACA). EGFR expression was associated with improved DSS in SCC (p = 0.012) but not for ACA. Co-expression of EGFR-HER3 was more likely in SCC then in ACA (p = 0.033). There was no correlation between outcome and any combination or clustering of biomarkers. Conclusions: The biomarkers p63 and Bcl-2 are predictive of DSS in NSCLC. EGFR expression is predictive of DSS in SCC. Sub-classification of NSCLC by histopathology is important as the relevance of some biomarkers (EGFR) would be lost if pooled. p63, Bcl-2, and EGFR may be used as prognostic markers in patients with NSCLC. Co-expression of EGFR-HER3 is more likely in SCC then in ACA. This may help explain the differential response to EGFR inhibitors in SCC versus ACA. No significant financial relationships to disclose.

Recent theories on second breakdown are reviewed. A simple concept on avalanche injection is proposed. The condition of distributed series resistance on stability factor analysis is discussed. An ingenious Motorola patent to realize this condition with low cost precision is illustrated and results on Power Base(R) product design shown. Observations confirm the substantial SOA improvement using above design features on the same emitter geometry and chip size. Conventional hot spot was not noted.