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BACKGROUND AND OBJECTIVES: We investigated a pertussis outbreak characterized by atypical cases, confirmed by polymerase chain reaction (PCR) alone at a single laboratory, which persisted despite high vaccine coverage and routine control measures. We aimed to determine whether Bordetella pertussis was the causative agent and advise on control interventions. METHODS: We conducted case ascertainment, confirmatory testing for pertussis and other pathogens, and an assessment for possible sources of specimen contamination, including a survey of clinic practices, sampling clinics for B pertussis DNA, and review of laboratory quality indicators. RESULTS: Between November 28, 2008, and September 4, 2009, 125 cases were reported, of which 92 (74%) were PCR positive. Cases occurring after April 2009 (n = 79; 63%) had fewer classic pertussis symptoms (63% vs 98%; P < .01), smaller amounts of B pertussis DNA (mean PCR cycle threshold value: 40.9 vs 33.1; P < .01), and a greater proportion of PCR-positive results (34% vs 6%; P < .01). Cultures and serology for B pertussis were negative. Other common respiratory pathogens were detected. We identified factors that likely resulted in specimen contamination at the point of collection: environmentally present B pertussis DNA in clinics from vaccine, clinic standard specimen collection practices, use of liquid transport medium, and lack of clinically relevant PCR cutoffs. CONCLUSIONS: A summer pertussis pseudo-outbreak, multifactorial in cause, likely occurred. Recommendations beyond standard practice were made to providers on specimen collection and environmental cleaning, and to laboratories on standardizing PCR protocols and reporting results, to minimize false-positive results from contaminated clinical specimens.

Purpose: We investigated the feasibility, safety, and biologic activity of adenovirus-mediated p53 gene transfer in patients with locally advanced bladder cancer. Patients and Methods: Patients with measurable, locally advanced transitional-cell carcinoma of the bladder who were not candidates for cystectomy were eligible. On a 28-day cycle, intravesical instillations of INGN 201 (Ad5CMV-p53) were administered on days 1 and 4 at three dose levels (1010 particles to 1012 particles) or on either 4 or 8 consecutive days at a single dose level (1012 particles). Results: Thirteen patients received a total of 22 courses without dose-limiting toxicity. Specific transgene expression was detected by reverse transcriptase polymerase chain reaction in bladder biopsy tissue from two of seven assessable patients. There were no changes in p53, p21waf1/cip1, or bax protein levels in bladder epithelium evident from immunohistochemical analysis of 11 assessable patients. Outpatient administration of multiple courses was feasible and well tolerated. A patient with advanced superficial bladder cancer showed evidence of tumor response. Conclusion: Intravesical instillation of Ad5CMV-p53 is safe, feasible, and biologically active when administered in multiple doses to patients with bladder cancer. Observations from this study indicate that this treatment has an antitumor effect in superficial transitional-cell carcinoma. Improvements in the efficiency of gene transfer and the levels of gene expression are required to develop more effective gene therapy for bladder cancer.

Advances in prevention and treatment of non-small cell lung cancer (NSCLC) are dependent in part on the characterization of tumor suppressor genes and oncogenes and the roles they elicit in the pathogenesis of this malignancy. Although v-ets erythroblastosis virus E26 oncogene homolog 2 (ETS2) is a canonical transcription factor that regulates various cancer-associated cellular and developmental processes including proliferation and migration, its function in lung carcinogenesis is still unknown. In this study we sought to examine the role of ETS2 in NSCLC pathogenesis. We first examined ETS2 mRNA expression in lung adenocarcinomas (n=80) and normal lung (n=30) which we profiled using microarrays, and in seven matched adenocarcinoma and normal lung pairs analyzed using next-generation sequencing technology. Both array and sequencing datasets revealed that ETS2 mRNA was significantly lower in lung adenocarcinomas relative to normal lung (p Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2177. doi:1538-7445.AM2012-2177

Background. The folate pathway is active in several cancers including malignant pleural mesothelioma (MPM). Membrane receptors folate receptor alpha (FRα), reduced folate carrier 1 (RFC1), proton coupled folate transporter (PCFT) and the enzyme thymidylate synthase (TS) regulate intra-cellular uptake of folate molecules and antifolate drugs. Due to the documented overexpression of folate markers in MPM they arise as potential targets for novel specific therapies with humanized monoclonal anti-FRα antibody and also to antifolate chemotherapy with pemetrexed. Material and Methods. We studied 79 surgically resected MPM (43 epithelioid, 28 biphasic and 13 sarcomatoid histology types). Protein expression of FRα, RFC1, PCFT and TS was examined by immunohistochemistry (IHC) in FFPE tissues placed in tissue microarray platform and results correlated to clinical-pathologic characteristics. Gene expression and DNA copy number changes of the FRα gene (FOLR1) were examined in 53 mesothelioma tumors using Affymetrix U133 Plus 2.0 microarray's and Illumina HumanOmni1-Quad v1.0 chips, respectively. The expression microarray data was analyzed in the GeneSpring GX 11 software (Agilent technologies Inc.) while DNA copy number changes were detected using the Nexus 5.0 software (BioDiscovery Inc.). Results. MPM frequently showed protein over-expression of FRα, RFC1 and PCFT. Nuclear TS expression positively correlated (P0) had a lower hazard ratio than those who did not (HR=0.309; 95%CI 0.159, 0.601; P=0.0005). Patients with any level (score >0) of cytoplasmic PCFT had lower hazard ratio than those with absence of the marker (HR=0.539; 95%CI 0.306, 0.952; P=0.0332). Our gene expression data analysis on 53 mesothelioma tumors showed greater than 2-fold increase in the FOLR1 and 3-fold increase in the TYMS messenger RNA levels compared to paired normal tissue. Additionally, at least 7 of these tumors show increased gene copy number at the FOLR1 locus (Ch11q 13.4). Conclusions. Folate membrane transporters (FRα, RFC1, PCFT) and the enzyme TS are frequently over expressed in MPM tumor tissues, and PCFT and TS are associated with tumors’ clinico-pathological features. The high level of expression of FRα in MPM supports the rational use of potential utilization of humanized monoclonal anti-FRα antibody and also standard antifolate chemotherapy in this disease. Supported by grant US DoD W81XWH-07-1-0306, ASCO CDA. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4121. doi:10.1158/1538-7445.AM2011-4121

Background: Most invasive lung adenocarcinomas (LAC) consist of mixed histological growth patterns including a non-invasive component referred as bronchioloalveolar carcinoma (BAC). Invasive LACs with BAC features are suspected to evolve from pure non-invasive (BAC) tumors. However, in this tumor type there is limited information about the molecular abnormalities associated to the progression from non-invasive to the invasive components. Materials and methods: To perform a detailed mapping analysis of molecular abnormalities comparing non-invasive and invasive sites in lung AC with BAC features we selected archival tumor tissue specimens from 77 LACs with more than 10% of non-invasive component out of 553 available tumors. Using whole histologic sections, we analyzed on each component (non-invasive and invasive) the immunohistochemical (IHC) expression of five markers known to be deregulated in lung AC, including EGFR, Her2-Neu, TTF-1, BRG1 and p53. We also compared in both tumor components the status of EGFR gene copy number gain (CNG) by fluorescent in situ hybridization and mutation of EGFR (exon 18-21) and KRAS (codons 12 and 13) by PCR-based sequencing using DNA extracted from microdissected tissue. For EGFR, we defined 2 types of CNGs: low (trysomy and low-polysomy), and high (high-polysomy and amplification). Results: Most cases had similar levels of expression in the non-invasive and invasive components of LAC for all IHC markers: EGFR, 54/70, 77%; Her2-Neu, 56/71, 79%; p53, 58/72, 80%; TTF-1, 57/71, 80%; and BRG1, 70/73, 96%. In a subset of cases, we found that the invasive component of LAC showed higher frequency of overexpression than non-invasive sites of EGFR (invasive 14/70, 20% vs. non-invasive 2/70, 3%) and Her2Neu (13/71, 18% vs. 2/71, 3%). EGFR mutations of exons 18-21 were found in 11/44 (20%) LAC, and in 7 (64%) mutant tumors the mutations were detected only in the invasive component. KRAS mutations were found in 14/56 (25%) tumors, and in all mutant tumors the mutations were found in both the non-invasive and corresponding invasive components. Any EGFR CNG was found in the invasive component of 20/56 (36%) tumors, including 9 cases with high CNG (4 high-polysomy and 5 gene amplification). In most (16/20, 80%) tumors the invasive component demonstrated higher CNG levels than the corresponding non-invasive sites. Interestingly, all 5 invasive tumors with EGFR amplification did not show such of abnormality in the corresponding non-invasive site. Conclusions: Our findings suggest that in LAC with BAC features the overexpression of EGFR and Her2Neu, and EGFR gene abnormalities (CNGs and mutations) are associated with the progression of non-invasive to invasive components of the tumors. In contrast, in LAC with KRAS mutation this abnormality commences at the non-invasive stage of tumor development. Supported by grant from Uniting Against Lung Cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1417. doi:10.1158/1538-7445.AM2011-1417

Background: Lung cancer is the leading cause of cancer-related death worldwide. Non-small cell lung cancer (NSCLC) accounts for ∼80% of all lung cancers. Brain metastases (BMs) are estimated to occur in up to 40% of patients with NSCLC. DNA copy number changes are common in cancer and lead to altered expression and function of genes residing within the affected region of the genome. The aim of this study was to compare DNA copy number abnormalities between primary tumors (PT) and their corresponding BM in NSCLC to identify genomic regions associated with BM development by using array comparative genomic hybridization (aCGH). Material and Method: From surgically resected NSCLC formalin-fixed and paraffin-embedded (FFPE) tissues, we selected 28 paired PTs and BMs (24 adenocarcinomas, and 4 squamous cell carcinomas, SCCs), and 55 additional BMs (43 adenocarcinomas, 2 SCCs, and 10 NSCLCs) specimens for analysis. Using DNA extracted from FFPE tumor tissues we performed oligo-based aCGH in 10 paired PT and BM adenocarcinomas. Statistical aberration detection algorithms of data were conducted by aCGH with Nexus Copy Number software v5.0. Immunohistochemical (IHC) analysis of two proteins (PSCA and LY6K) coded by genes located in 8q24.3 region was performed in both PT and BM specimens. Results: In the aCGH analysis, multiple chromosomal regions with significant (P≤0.05) copy number gain and loss were detected. Whereas PT demonstrated 32 regions (harboring 933 genes) of copy number gain and 16 regions (108 genes) of copy number loss, BM showed 31 regions (1274 genes) with gains and 16 regions (90 genes) with losses. BM demonstrated significantly (P≤0.01) higher frequency of copy number gains in 3 chromosomal regions: 8q24.3 (BM 100% vs. PT 40%), 19q13.33 (BM 90% vs. PT 20%), and 20q13.12 (BM 60% vs. PT 0%). The 8q24.3 region with copy number gain in tumors harbors 7 genes, including Prostate Stem Cell Antigen (PSCA) and Lymphocyte Antigen 6 Complex Locus K (LY6K). Interestingly, in our IHC analysis, BMs (n=84; mean score 141.8) showed a significantly (P=0.008) higher score of expression of PSCA than the corresponding PTs (n=27; mean score 97.8). Similar significant differences were detected when we analyzed only adenocarcinomas samples. No significant differences in the level of IHC expression was detected comparing BMs (n=83; mean score 64) and PTs (n=27; mean score 68.5) for LY6K. Conclusions: By aCGH and subsequent protein expression analysis we have identified that PSCA may play an important role in the development of NSCLC brain metastasis. Similarly to prostate cancer, PSCA represents a novel potential marker for NSCLC metastasis and therapeutic target for advanced lung tumors. Supported by grant US DoD W81XWH-07-1-0306 and Jimmy L Hewlett Foundation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 5166. doi:10.1158/1538-7445.AM2011-5166

Non-small cell lung cancer (NSCLC) represents the majority (85%) of lung cancers and is comprised mainly of adenocarcinomas and squamous cell carcinomas (SCCs). The sequential pathogenesis of lung adenocarcinomas and SCCs occurs through dissimilar phases as the former tumors typically arise in the lung periphery whereas the latter normally arise near the central airway. We assessed the expression of SOX2, an embryonic stem cell transcriptional factor that also plays important roles in the proliferation of basal tracheal cells and whose expression is restricted to the main and central airways and bronchioles of the developing and adult mouse lung, in NSCLC by various methodologies. Here, we found that SOX2 mRNA levels, from various published datasets, were significantly elevated in lung SCCs compared to adenocarcinomas (all p Running title: SOX2 abnormalities in NSCLC 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 5166.

Background: Non-small cell lung cancer (NSCLC) represents the majority (85%) of lung cancers and is comprised mainly of adenocarcinomas and squamous cell carcinomas (SCCs). The sequential pathogenesis of lung adenocarcinomas and SCCs occurs through dissimilar phases as the former tumors typically arise in the lung periphery whereas the latter normally arise near the central airway. Methodology/Principal Findings:We assessed the expression of SOX2, an embryonic stem cell transcriptional factor that also plays important roles in the proliferation of basal tracheal cells and whose expression is restricted to the main and central airways and bronchioles of the developing and adult mouse lung, in NSCLC by various methodologies. Here, we found that SOX2 mRNA levels, from various published datasets, were significantly elevated in lung SCCs compared to adenocarcinomas (all p, published_or_final_version

Introduction:Folate receptor alpha (FRα) and reduced folate carrier-1 (RFC1) regulate uptake of folate molecules inside the cell. FRα is a potential biomarker of tumors response to antifolate chemotherapy, and a target for therapies using humanized monocloncal antibody. Information on the protein expression of these receptors in non–small-cell lung carcinoma (NSCLC) is limited.Material and Methods:Expressions of FRα and RFC1 were examined by immunohistochemistry (IHC) in 320 surgically resected NSCLC (202 adenocarcinomas and 118 squamous cell carcinomas) tissue specimens and correlated with patients’ clinico-pathologic characteristics. Folate receptor α gene (FOLR1) mRNA expression was examined using publicly available microarray datasets. FRα expression was correlated with thymidylate synthase and p53 expression in NSCLCs, and with epidermal growth factor receptor (EGFR) and V-Ki-ras2 Kirsten rat sarcoma viral (KRAS) gene mutations in adenocarcinomas.Results:NSCLC overexpressed FRα and RFC1. In a multivariate analysis, lung adenocarcinomas were more likely to express FRα in the cytoplasm (OR = 4.39; p < 0.0001) and membrane (OR = 5.34; p < 0.0001) of malignant cells than squamous cell carcinomas. Tumors from never-smokers were more likely to express cytoplasmic (OR = 3.35; p