Supplementary Table 1 from Antitumor Activity of an Oncolytic Adenovirus-Delivered Oncogene Small Interfering RNA

The National Survey on Drug Use and Health (NSDUH), an annual survey of the U.S. civilian, noninstitutionalized population aged 12 or older, is a major source of substance use and mental illness data. The 2010 estimates were produced using weights poststratified to 2010 population control totals (intercensal estimates) derived from the 2000 decennial census; however, the 2011 estimates were produced using weights poststratified to 2011 population control totals derived from the 2010 decennial census. This study was done to determine whether the change in the source of the control totals had an effect on the level of change observed between the 2010 and the 2011 estimates. To examine this "census effect," 2010 estimates were also produced using weights poststratified to 2010 population control totals derived from the 2010 decennial census, resulting in two sets of weights for use on the 2010 data. NSDUH estimates were compared using both sets of 2010 estimates along with the 2011 estimates. Substance use estimates were more affected by the census effect than were mental illness estimates, and they were more notable for estimated totals compared with rates.

Different to the conventional visible‐CCFL backlight, UV excited flat lighting (UFL) applies UV‐CCFL to irradiate 254 nm wavelengths to excite extrinsic phosphor layer to perform uniform and flat lighting for LCD‐TV applications. UFL can yield longer life time due to less phosphor damage. Additionally, high color rendering, minor color deviation, wider FWHM (Full width at half maximum) angle, and better lamp mura relaxation, were demonstrated to achieve a high quality display with slim backlight module.

MicroRNAs (miRNAs) are a recently discovered class of small (approximately 18-24 nt) nucleic acids that negatively regulate gene expression. This novel class of molecules modulates a wide array of growth and differentiation processes in human cancers. High throughput analyses, utilizing the solid phase, array platform, or liquid phase, bead-based hybridization have variously demonstrated that miRNA expression was commonly dysregulated in human cancer. miRNA expression profiling has shown promise in defining malignant status in retrospective studies. Considerable disagreement remains with respect to the miRNA signature for a specific cancer cell type, which appears to depend largely on the analytical platform. Nonetheless, various internally controlled studies have successfully identified the histotype of tumors of unknown origin according to miRNA expression profile. The evaluation of miRNAs expression may also be of prognostic value, as best exemplified by the correlation of let-7 and mir-155 levels with disease survival in nonsmall cell lung cancer.

Abstract Despite successes in animal models, cancer gene therapy with small interfering RNAs (siRNA) is hindered by the lack of an optimal delivery platform. We examined the applicability of the replication-competent, oncolytic adenovirus, ONYX-411, to deliver a mutant K-ras siRNA transgene to human cancer cells. Proof-of-principle studies showed an additive tumor growth–inhibitory response through siRNA-mediated K-ras knockdown and ONYX-411-mediated cancer cell lysis. A novel construct, termed Internavec (for interfering RNA vector), was generated by cloning a K-rasv12-specific siRNAras-4 hairpin construct under the control of the human H1 promoter into the deleted E3b region of ONYX-411. Internavec acquired an increase in potency of ∼10-fold in human cancer cells expressing the relevant K-rasv12 mutation (H79, H441, and SW480), as defined by a reduction in the effective dose needed to achieve 50% growth inhibition (ED50). Internavec remained attenuated in nonmalignant epithelial cells. Daily intratumoral injections of Internavec (five daily injections of 1 × 108 plaque-forming units) significantly reduced the growth of s.c. H79 pancreatic cancer xenografts in nu/nu mice by 85.5%, including complete growth suppression in three of five mice. Parental ONYX-411 or ONYX-411-siRNAGFP was markedly less effective (47.8% growth reduction, P = 0.03; and 44.1% growth reduction, P = 0.03, respectively). siRNAras transgene activity contributed to cell cycle blockage, increased apoptosis, and marked down-regulation of Ras signaling–related gene expression (AKT2, GSK3β, E2F2, and MAP4K5). These findings indicate that Internavec can generate a two-pronged attack on tumor cells through oncogene knockdown and viral oncolysis, resulting in a significantly enhanced antitumor outcome. (Cancer Res 2006; 66(19): 9736-43)

Small interfering RNAs (siRNA) are small double stranded RNA molecules that mediate specific and highly potent post- transcriptional gene knockdown. siRNA-mediated knockdown of relevant genes resulted in tumor growth inhibition in a variety of human cancer cell types. However, clinical applicability of this technology has been hindered by the lack of an optimal delivery platform. We examined the use of the conditional replicative, oncolytic virus ONYX-411 to deliver an siRNA that knocks down the K-rasv-12 mutant oncogene overexpressed in many cancer cells. ONYX-411 carries an E1A deletion and E2F-1 conditional promoters in the E1 and E4 regions, hence limiting replication to cancer cells with a defective retinoblastoma tumor suppressor protein (pRB) pathway and E2F-1 overexpression. Viral oncolysis and replication lead to tumor cell reinfection, which is expected to amplify siRNA-knockdown effects. In proof-of-principle studies, we observed additive tumor growth inhibitory responses through siRNA-mediated K-ras knockdown and ONYX-411 infection. siRNA expression at the late infectious phase did not negatively impact viral replicative activity. A novel construct (Internavec) was generated by placing the K-rasv-12-specific siRNAras-4 hairpin construct under the control of endogenous, viral E3B late gene regulatory elements of ONYX-411. When tested against a panel of viral permissive human cancer lines (H79, H441, SW480, H522, H596), Internavec displayed a significantly elevated anti-tumor response (as compared with parental ONYX-411) only in host cells expressing the relevant K- rasv-12 mutation. The required Internavec dose (MOI of 1.03) for attaining 50% tumor cell growth inhibition (ED50) was 11-fold- lower than parental ONYX-411 (MOI of 11.6) and 14x lower than the control construct containing an siRNA against the green fluorescence protein (ONYX-411-siRNAGFP, MOI of 14.7). By comparison, Internavec remains attenuated in non-malignant HMEC cells. Hence the restricted viral activity within the tumor microenvironment can potentially serve to safeguard against previously reported "off target" siRNA effects. Daily intratumoral injections with Internavec (5 @ 1×108 pfu) significantly reduced H79 pancreatic cancer xenograft growth by 85.5 %, including complete growth suppression in 3 of 5 mice. Parental ONYX-411 or ONYX-411-siRNAGFP was markedly less effective (47.8% growth reduction, p=0.03; and 44.1% growth reduction, p=0.03, respectively). These findings indicate that Internavec can generate a two-pronged attack on tumor cells through oncogene knockdown and viral oncolysis, leading to significantly enhanced antitumor outcomes.

Arthritis and osteonecrosis affect a large number of patients with systemic lupus erythematosus (SLE). A patient with history of SLE suffered a traumatic fracture of the left foot. Despite a long period of immobilization and internal fixation, the fracture failed to heal and required arthrodeses with removal of the phalanx. Histopathological investigation revealed destruction of cartilage, subchondral cystic degeneration, vasculitis, deposition of fibrinogen, type III collagen and fibronectin, absence of bone remolding, and detectable F-actin. The nonhealing was therefore due to lack of progression of healing process beyond the initial stage. There was deposition of immunoglobulins and complement C4b, possibly forming immune complex by autoantibodies and cellular components. The authors found that MSE55 protein, required for polymerization of actin and initiation of cellular process organization, had a similar cellular deposition as that of immunoglobulins. Autoantibodies thus may inhibit differentiation of the bone cells, and resulted in nonunion in the patient.

Objectives.The objective was to evaluate the sensitivity and specificity of cervical cytology in women infected with the human immunodeficiency virus (HIV), risk factors for abnormal cytology in HIV-infected and uninfected women, and risk factors for histologic diagnosis of cervical intraepithelial neoplasia (CIN) in HIV-infected women.Methods.Methods included a cross-sectional analysis of cervical cytology, colposcopic impression, and histology in 248 HIV-infected women and multivariate analyses of risk factors for abnormal cytology in 253 HIV-infected and 220 uninfected women and risk factors for CIN in 186 HIV-infected women.Results.The sensitivity and specificity of cytology for all CIN grades were 0.60 and 0.80 and, for high-grade CIN, 0.83 and 0.74. The prevalence of abnormal cytology was 32.9% in HIV-infected and 7.6% in HIV-negative women. Independent risk factors for abnormal cytology were immunodeficiency [odds ratio (OR) 8–17,P< 0.001] and human papillomavirus (HPV) infection (OR = 5,P< 0.001). The prevalence of CIN on histology was 32% in HIV-infected women, and the only independent risk factor for CIN was oncogenic HPV type (OR = 5,P= 0.005).Conclusion.Given the high prevalence of abnormal cytology and CIN in HIV-infected women, cytologic screening has significant limitations. Both immunodeficiency and type of HPV infection are important risk factors.

From the Arnold & Marie Schwartz College of Pharmacy (RS), the Department of Pathology (PC) and AIDS Unit (MK), State University of New York Health Science Center at Brooklyn, New York

BACKGROUND: Studies demonstrating a high prevalence of abnormal Pap smears and cervical intraepithelial neoplasia (CIN) in HIV-infected women have led to concerns about appropriate surveillance measures. The objective of this evaluation was to determine the specificity and sensitivity of cervical cytology in a large group of HIV-infected women. METHODS: From 1991 to 1994, 285 HIV-infected women were enrolled in a prospective cohort study of the natural history of HIV disease and associated gynecologic manifestations. The baseline examination included cervical cytology, colposcopy and colposcopically directed biopsy. If no lesion was visualized colposcopically, a biopsy was taken of the transformation zone. All cytology slides were reviewed by a senior cytologist after initial evaluation. All histopathology slides were reviewed by an experienced pathologist blinded to the woman's HIV status. RESULTS: Among 249 HIV-infected women who had cervical cytology, colposcopy and cervical biopsy, 32.4% had abnormal cytology, including 8.8% with atypical squamous cells of undetermined significance (ASCUS), 15.6% with low grade squamous intraepithelial lesions (SIL) and 8.0% with high grade SIL. On colposcopic visualization, 75% had lesions consistent with CIN or condylomatous lesions. Histology showed CIN1 in 19.3%, CIN2 in 7.6%, and CIN3 in 4.0%. The sensitivity and specificity of cytology for CIN of all grades were 0.60 and 0.80 respectively. For high grade CIN, the sensitivity and specificity were 0.83 and 0.74. The false negative rate of normal smears was 18% with 5 cases of CIN 2-3 and 26 of CIN 1. All women with CIN 2-3 had either an abnormal smear or a lesion on colposcopy. However, 15% of women with normal smears and no colposcopic evidence of CIN had CIN 1 on biopsy. CONCLUSION: Cervical cytology in HIV-infected women has sensitivity and specificity within the range reported in normal populations. However, the high prevalence of CIN in the immunodeficient suggest that colposcopy or other tests should supplement cytology because of its inherent limited sensitivity. Studies of the potential for progression of CIN in HIV-infected women are needed to resolve questions on appropriate screening protocols.

We report a case of recurrent intravascular papillary endothelial hyperplasia.

The purpose of this study was to compare cytology and colposcopy as predictors of cervical intraepithelial neoplasia (CIN) in women infected with the human immunodeficiency virus (HIV). A cross-sectional analysis of cytology, colposcopy, and colposcopic biopsy results from 51 HIV-seropositive women attending an ambulatory HIV service was conducted. Cytology slides were reviewed by two cytopathologists blinded to patients' HIV status. There was strong agreement in the readings of two cytopathologists, with a kappa score of 0.9. Of 29 women with normal cytology, 21 (72%) had pathology on histology, including 7 (24%) with CIN. Colposcopic impression correlated well with histology results. Of 22 women with abnormal cytology, 82% had abnormal histology. The overall prevalence of CIN was high at 45%, increasing from 35% in women with CD4 counts over 400 to 56% in women with CD4 counts below 200. In conclusion, screening cytology is limited by false-negative results; routine colposcopy should be considered in this high-risk population.

The survival rates of 117 black and white patients treated by primary radiation for carcinoma of the prostate at the State University of New York Health Science Center at Brooklyn and Kings County Hospital Center were analyzed according to age and race. In addition, stage, grade, and delay time in seeking medical attention were analyzed. Survival was similar in both young (<60 years) and old (≥60 years) patients, with 45% and 41% 5-year-survival rates, respectively. Survival was better in white patients, 48% 5-year survival, than in blacks, 35% 5-year survival. Black patients presented with higher stage disease than whites (p < 0.01). This trend was even greater in young black males, who had higher grade (64%. versus 11%; p < 0.04) and higher stage tumors (p < 0.05). In addition, young blacks delayed seeking medical attention >3 months 72% of the time, as compared to 0% in white young males (p < 0.005). A survival difference was also seen in young blacks as compared with young whites: 3.9-year median survival versus 6.0-year median survival, respectively.

The survival of 117 black and white patients treated by radiation for carcinoma of the prostate at SUNY Health Science Center at Brooklyn (SUNY/HSCB) and Kings County Hospital Center (KCHC) was analyzed according to Gleason's grading system. The effect of total pattern score and its relationship to stage and survival and to race were intercompared. In both black and white racial groups, there was strong correlation between high pattern score and high stage p = less than 001. The percentage of black patients presenting with high pattern score (7-10) was significantly greater, 43 versus 27%; this adversely affected stage and survival. The median survival for white and black patients was 4.8 and 3.2 years, respectively; p = 0.007. Stage for stage and grade for grade, survival was similar in both racial groups.