Combinatorial strategies based on CRAd-IL24 and CRAd-ING4 virotherapy with anti-angiogenesis treatment for ovarian cancer

BACKGROUND: A major hurdle incurrent to the human clinical application of conditionally replicative adenovirus (CRAd)-based virotherapy agents is their limited therapeutic efficacy. In this study we evaluated whether arming our previously reported Ad5/3Δ24 CRAd vector containing a 24-base pair deletion in the E1A conserved region 2, which allows selective replication within Rb-p16-deficient tumor cells, to express therapeutic genes could improve oncolytic virus potency in ovarian cancer cells. We choose to assess the therapeutic benefits achieved by virus-mediated expression of interleukin 24 (IL-24), a cytokine-like protein of the IL-10 family, and the inhibitor of growth 4 (ING4) tumor suppressor protein. RESULTS: The generated CRAd-IL24 and CRAd-ING4 vectors were tested in ovarian cancer cell lines in vitro to compare their replication, yield, and cytotoxic effects with control CRAd Ad5/3∆24 lacking the therapeutic gene. These studies showed that CRAd-IL24 infection resulted in significantly increased yield of infectious particles, which translated to a marked enhancement of virus-induced cytotoxic effects as compared to CRAd-ING4 and non-armed CRAd. Testing CRAd-IL24 and CRAd-ING4 vectors combined together did not revealed synergistic effects exceeding oncolytic potency of single CRAD-IL24 vector. Both CRAds were also tested along with anti-VEGF monoclonal antibody Avastin and showed no significant augmentation of viral cytolysis by anti-angiogenesis treatment in vitro. CONCLUSIONS: Our studies validated that arming with these key immunomodulatory genes was not deleterious to virus-mediated oncolysis. These findings thus, warrant further preclinical studies of CRAd-IL24 tumoricidal efficacy in murine ovarian cancer models to establish its potential utility for the virotherapy of primary and advanced neoplastic diseases

Quality of Life in Patients with Oral Potentially Malignant Disorders: Oral Lichen Planus and Oral Epithelial Dysplasia

Background Oral potential malignant disorders (OPMDs) are lesions and conditions that increase the risk for malignant transformation. OPMDs have an impact on patients’ health status and Quality of life (QoL). The aim of our study is to evaluate and compare QoL in patients with oral lichen planus and oral epithelial dysplasia as a setting of clinical leukoplakia. Methods This is a cross-sectional study with a sample size of 100 patients divided into, 53 subjects with Oral Lichen Planus (53.0%), 39 patients with oral epithelial dysplasia as a setting of solitary leukoplakia (39.0%), and 8 subjects with oral lichen planus with oral epithelial dysplasia (8.0%). Patients who fit the criteria were asked to fill out three different questionnaires. 26-item Chronic Oral Mucosal Disease Questionnaire (COMDQ-26), Hospital Anxiety and Depression Scale (HADS), and the Oral potentially malignant disorders QoL questionnaire (OPMDsQoL). Results Both the COMD-26 and OPMDQoL questionnaire scores were significantly different across the three disease groups. Relative to patients aged \u3e65, the 40-64 age group added 6.8 points to the COMD-26 survey score (P \u3c 0.05). Relative to oral epithelial dysplasia, oral lichen planus added 15.7 points to the COMD-26 survey score (P \u3c 0.001). Relative to oral epithelial dysplasia, oral lichen plus added 8.9 points to the OPMDQoL survey score (P = 0.003). Conclusion Within the limitation of our study OLP shows significant poorer QoL in compared to OED as a setting of clinical OL. Younger individuals affected with OLP, and OED showed significant impact in QoL in compared to older individuals

Virological diagnosis of dengue fever in Jeddah, Saudi Arabia: Comparison between RT-PCR and virus isolation in cell culture

A total of 233 serum samples were collected from patients presenting to King Abdulaziz University Hospital with suspected cases of Dengue Fever (DF) from 2006 to 2008. Dengue virus was successfully isolated from 70 samples by culture on C6/36 and LLC-MK2 cells; it was then detected by indirect immunofluorescence assay (IFA). The cytopathic effect (CPE) of dengue virus on C6/36 appeared in most of the samples within 1-4 days post-inoculation comparing to 7-12 days on LLC-MK2 cells, and this was characterized by the ability to induce syncytia and multinucleated giant cells. On the other hand, by using RT-PCR technique, 87 (37.3%) samples were positive. All 70 (30.4%) samples with positive cell culture results were detectable by RT-PCR in addition to 17 culture-negative samples were RT-PCR positive. Dengue virus type 1 (DENV-1) was the dominant serotype followed by DENV- 3 and DENV-2, while DENV-4 was not detected in tested samples. These results indicate that DENV-RNA detection by RT-PCR is more sensitive than virus isolation. We suggest that the high sensitivity coupled with the turnaround time, have made the RT-PCR a better choice as a routine test for DENV diagnosis

Defining a murine ovarian cancer model for the evaluation of conditionally-replicative adenovirus (CRAd) virotherapy agents

Abstract Background Virotherapy represents a promising approach for ovarian cancer. In this regard, conditionally replicative adenovirus (CRAd) has been translated to the context of human clinical trials. Advanced design of CRAds has sought to exploit their capacity to induce anti-tumor immunization by configuring immunoregulatory molecule within the CRAd genome. Unfortunately, employed murine xenograft models do not allow full analysis of the immunologic activity linked to CRAd replication. Results We developed CRAds based on the Ad5/3-Delta24 design encoding cytokines. Whereas the encoded cytokines did not impact adversely CRAd-induced oncolysis in vitro, no gain in anti-tumor activity was noted in immune-incompetent murine models with human ovarian cancer xenografts. On this basis, we explored the potential utility of the murine syngeneic immunocompetent ID8 ovarian cancer model. Of note, the ID8 murine ovarian cancer cell lines exhibited CRAd-mediated cytolysis. The use of this model now enables the rational design of oncolytic agents to achieve anti-tumor immunotherapy. Conclusions Limits of widely employed murine xenograft models of ovarian cancer limit their utility for design and study of armed CRAd virotherapy agents. The ID8 model exhibited CRAd-induced oncolysis. This feature predicate its potential utility for the study of CRAd-based virotherapy agents