Citations

Abstract The terminology for human papillomavirus (HPV)–associated squamous lesions of the lower anogenital tract has a long history marked by disparate diagnostic terms derived from multiple specialties. It often does not reflect current knowledge of HPV biology and pathogenesis. A consensus process was convened to recommend terminology unified across lower anogenital sites. The goal was to create a histopathologic nomenclature system that reflects current knowledge of HPV biology, optimally uses available biomarkers, and facilitates clear communication across different medical specialties. The Lower Anogenital Squamous Terminology (LAST) Project was cosponsored by the College of American Pathologists and the American Society for Colposcopy and Cervical Pathology and included 5 working groups; 3 work groups performed comprehensive literature reviews and developed draft recommendations. Another work group provided the historical background and the fifth will continue to foster implementation of the LAST recommendations. After an open comment period, the draft recommendations were presented at a consensus conference attended by LAST work group members, advisors, and representatives from 35 stakeholder organizations including professional societies and government agencies. Recommendations were finalized and voted on at the consensus meeting. The final, approved recommendations standardize biologically relevant histopathologic terminology for HPV-associated squamous intraepithelial lesions and superficially invasive squamous carcinomas across all lower anogenital tract sites and detail the appropriate use of specific biomarkers to clarify histologic interpretations and enhance diagnostic accuracy. A plan for disseminating and monitoring recommendation implementation in the practicing community was also developed. The implemented recommendations will facilitate communication between pathologists and their clinical colleagues and improve accuracy of histologic diagnosis with the ultimate goal of providing optimal patient care.

Abstract Metastatic malignant melanoma is an incurable disease with average 5-year survival rates for patients with disseminated disease ranging from 5-18%. Despite the recent development of promising targeted and immunological approaches for treatment of melanoma patients, there remains an urgent need to improve the effectiveness of conventional chemotherapeutics such as TMZ treatment for those patients ineligible or unresponsive to these approaches. We hypothesized that genetic alterations or contexts of vulnerability (COVs) exist in melanoma cells that mediate sensitivity to TMZ, and thus developed RNA-interference-based (RNAi) TMZ chemosensitization assays in melanoma cell lines in order to better understand how to pharmacologically augment TMZ response. First, we extensively developed RNAi sensitization assay in 17 melanoma cell lines followed by screening siRNA kinome library in these cell lines. This screening result, combined with our previous melanoma genomic data (gene expression, aCGH, SNP, and gene mutations), would prioritize the critical essential genes in melanoma. In addition, we profiled 17 melanoma cell lines for TMZ sensitivity and subsequently optimized RNAi screening parameters for detection of TMZ sensitization in one sensitive (UACC903) and four resistant (UACC647, UACC1308, UACC91 and UACC1940) cell lines. We then performed high-throughput RNAi screens with a 7,000 gene siRNA library targeting the druggable genome in these five cell lines. The data from the screens were rigorously evaluated and top TMZ-sensitizing hits were identified and confirmed. Based on these data, we prioritized the ATR/ATM/Chk1 pathway for further evaluation. We tested two Chk1 inhibitors for synergistic effects when combined with TMZ. Notably, both Chk1 inhibitors demonstrated good synergistic effects in the TMZ-resistant cell lines, with moderate/marginal effects in the sensitive line UACC903. These data are consistent with previous reports that inhibition of Chk1 can sensitize TMZ-induced cytotoxicity in glioma cells, and suggest that targeting this pathway in melanoma may augment TMZ response in melanoma. We are presently following up these results across a larger panel of melanoma cell lines, as well as evaluating pharmacological inhibitors of additional TMZ-sensitizing siRNA hits and pathways with the hope that these results may serve as a starting point for the development of novel combination therapies for melanoma patients. 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 5457. doi:10.1158/1538-7445.AM2011-5457

The association between bicuspid aortic valve (BAV) and ascending aorta aneurysm is well described. Replacement of the ascending aorta is now being considered at 4.5 cm. We identified patients confirmed with BAV who underwent elective aortic valve replacement (AVR) with a mechanical St Jude Bioprosthesis from 1994 to 2000 who were ≤65 years of age at the time of surgery. Follow-up imaging was obtained by computed tomography (CT) angiography or echocardiography. A total of 225 patients who underwent AVR were identified; 60 patients had a BAV. Of all, 36 (60%) patients with BAV returned for follow-up imaging of their ascending aorta. Eight patients (22%) had diameters classifiable as aneurysmal (>4.5 cm) that developed within 9.6 ± 4.1 years from implant and requiring surgery. Of all, 7 patients (12%) died within 5.9 ± 2.5 years from their implant date. Lifelong serial monitoring of the ascending aorta for patients with BAV should be the standard of care.

Oncolytic reovirus is carried by cells to tumors and protected from neutralizing antibodies in the circulation.

Background —Although prompt treatment is a cornerstone of the management of acute myocardial infarction (AMI), prior studies have shown that one fourth of AMI patients arrive at the hospital >6 hours after symptom onset. It would be valuable to identify individuals at highest risk for late arrival, but predisposing factors have yet to be fully characterized. Methods and Results —Data from the Cooperative Cardiovascular Project, involving Medicare beneficiaries aged >65 years hospitalized between January 1994 and February 1996 with confirmed AMI, were used to identify patients who presented “late” (≥6 hours after symptom onset). Patient characteristics were tested for associations with late presentation by use of backward stepwise logistic regression. Among 102 339 subjects, 29.4% arrived late. Significant predictors of late arrival (odds ratio, 95% CI) included diabetes (1.11, 1.07 to 1.14) and a history of angina (1.32, 1.28 to 1.35), whereas prior MI (0.82, 0.79 to 0.85), prior angioplasty (0.80, 0.75 to 0.85), prior bypass surgery (0.93, 0.89 to 0.98), and cardiac arrest (0.52, 0.46 to 0.58) predicted early presentation. Additionally, initial evaluation at an outpatient clinic (2.63, 2.51 to 2.75) and daytime presentation (1.67, 1.59 to 1.72) predicted late arrival. Finally, female sex, black race, and poverty, which were evaluated with an 8-level race–sex–socioeconomic status interaction term, were also risk factors for delay. Conclusions —Delayed hospital presentation is a common problem among Medicare beneficiaries with AMI. Factors associated with delay include not only clinical and logistical issues but also race, sex, and socioeconomic characteristics. Education efforts designed to hasten AMI treatment should be directed at individuals with risk factors for late arrival.

Background This study was undertaken to test the hypothesis that β-adrenergic stimulation in the setting of membrane depolarization will potentiate flecainide-induced conduction slowing. Methods and Results To elucidate the potential mechanism for the flecainide proarrhythmia observed in CAST, the voltage dependence of β-adrenergic modulation of impulse propagation in eight flecainide-superfused canine Purkinje fibers was examined with a dual-microelectrode technique. At physiological membrane potentials (V m ) ([K + ] o =5.4 μmol), 1 μmol flecainide decreased V̇ max from 698±55 to 610±72 V/s ( P =.003) and squared conduction velocity (θ 2 ) from 2.11±1.1 to 1.72±0.9 (m/s) 2 ( P =.001). With K + depolarization to V m =−70 mV, flecainide further reduced V̇ max from 306±101 to 245±65 V/s and θ 2 from 1.12±0.4 to 0.99±0.6 (m/s) 2 , producing a 2.0-mV hyperpolarizing shift of apparent Na + channel availability curves derived from θ 2 . The addition of 1 μmol isoproterenol to flecainide-superfused fibers at physiological V m increased θ 2 by 8% to 1.84±0.6 (m/s) 2 ( P <.01) without altering V̇ max . At −70 mV, the addition of isoproterenol magnified the flecainide-induced reduction of V̇ max an additional 24% to 185±52 V/s ( P <.01) and θ 2 by 17% to 0.82±0.5 (m/s) 2 ( P =.04), producing an additional 1.8-mV ( P =.002) and 1.9-mV ( P =.002) hyperpolarizing shift in the apparent Na + channel inactivation curves generated from V̇ max and θ 2 , respectively. At physiological V m , the action potential duration (APD 95 ) was reduced from 307±35 to 269±27 ms ( P <.001) by flecainide and subsequently to 217±4 ms ( P <.001) with isoproterenol addition. With 12 mmol/L K + , APD 95 decreased from 198±23 to 182±17 ms ( P =.005) with flecainide and to 164±10 ms ( P =.004) with isoproterenol. Conclusions At depolarized V m , isoproterenol amplified the flecainide-induced reduction of V̇ max and θ 2 , suggesting a further adrenergic-mediated reduction of Na + current. Consequently, the synergy between catecholamines and flecainide at depolarized V m and the shortened APD 95 could facilitate arrhythmogenesis in the presence of underlying ischemia.

AbstractVEGF induces vascular permeability (VP) in ischemic diseases and cancer, leading to many pathophysiological consequences. The molecular mechanisms by which VEGF acts to induce hyperpermeability are poorly understood and in vivo models that easily facilitate real-time, genetic studies of VP do not exist. In the present study, we report a heat-inducible VEGF transgenic zebrafish (Danio rerio) model through which VP can be monitored in real time. Using this approach with morpholino-mediated gene knock-down and knockout mice, we describe a novel role of phospholipase Cβ3 as a negative regulator of VEGF-mediated VP by regulating intracellular Ca2+ release. Our results suggest an important effect of PLCβ3 on VP and provide a new model with which to identify genetic regulators of VP crucial to several disease processes.