Abstract Introduction. Cancer 101, a cancer education program, is a critical resource to increase cancer knowledge, awareness, and cancer prevention activities to improve cancer health and survival rates. The purpose of this study was to test the feasibility of delivering the Cancer 101 curriculum to incarcerated adult males to improve cancer knowledge and increase cancer prevention activities. Methods. Cancer 101 curriculum was pilot tested for adaptation with representatives from the target audience in a New York and Pennsylvania prison facilities and based on their feedback modifications were implemented. Three prison facilities participated in adapting Cancer 101 for this study; participation in the Cancer 101 educational intervention was limited to individuals who were current smokers or former smokers. Pre-test and post-test surveys were used to assess knowledge of, perceived attitudes regarding and benefits of cancer prevention activities at baseline and immediately after completing the Cancer 101 educational intervention. To determine whether cancer knowledge improved after participating in the Cancer 101 program, a paired t-test procedure was conducted; since improvement was expected, a one-tailed alpha of .05 was specified. Results. The mean cancer knowledge scores differed prior to and after completing the Cancer 101 intervention, t(165) = -14.65, p < .000. Mean cancer knowledge scores prior to the Cancer 101 intervention (M = 28.42, SD = 4.21) were significantly lower than after the intervention (M = 32.68, SD = 3.36). Therefore, the intervention had a positive effect on cancer knowledge. Participants also reported that the primary reasons for participating in the Cancer 101 intervention were to develop knowledge about cancer and the skills to participate in cancer prevention activities and to educate other inmates and family members about cancer. Participants demonstrated significant increases in cancer knowledge and interest in partaking in cancer prevention activities. Discussion. A prison-based Cancer 101 educational intervention provides opportunities for inmates to increase cancer knowledge, as well as promote action for cancer control during incarceration. Citation Format: Pamela Valera, Daniel Mackey, Zi Lian, William McLaughlin. The feasibility of delivering a cancer education program in correctional facilities [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-179.

Abstract Tumor irradiation during cancer treatment induces cell death by apoptosis, mitotic death, and clonogenic death. With expanded global access to cancer radiotherapy, there is a need to discover agents with anti-cancer and radiation sensitizing properties. in vitro high-throughput screening (HTS) systems routinely used to screen agents for cytotoxicity assays unfortunately do not directly measure clonogenic potential and fail to accurately predict the efficacy of an agent either in subsequent preclinical animal model testing or in clinical trials. Clonogenic assays unlike colorimetric and metabolic assays for determining cell viability and apoptosis, are less susceptible to artifacts. We are addressing these requirements through an integrated I-PARTS platform development and optimization. Here, the design and the prototype of a fully enclosed imaging/X-ray irradiation system will be presented. The proof of principle experimentation to obtain high content information on the cellular response to specific primary human cells (normal or tumors) to a variety of radiation doses and fraction modality will be described. The initial approaches taken towards the establishment of an integrated platform to screen anti-cancer drugs will be discussed. Citation Format: Rao V. Papineni, Sylvain Costes, Shayoni Ray, William McLaughlin, Brendan Lukaniec, Kurt Kruger, Keith Kanugi. Design and modular development of I-PARTS: An integrated platform for anti-cancer radiation therapeutics screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-070.

Introduction Nearly nine million Americans pass through the criminal justice system (i.e., prison, jail, parole, and probation). Several reports have indicated that approximately 60-80% of inmates in the U.S. have histories of tobacco use, compared to 30% of the general population. This is significant, because, despite declining smoking rates in the general population, tobacco use is still the leading cause of preventable mortality and morbidity in the nation. Moreover, tobacco use by inmates has not been the focus of cancer health studies. That being said, this study describes the implementation strategy to support the adoption of a cancer health disparities education program, called Cancer 101 aimed at improving cancer health among underserved populations. Methods/Design Cancer 101 will be offered to three male correctional facilities in New York and Pennsylvania using a stepped wedge design. Due to adopting Cancer 101 in a real-world environment with plenty of logistical and structural constraints, the order in which different cohorts receive the intervention will be determined at random; however, all groups will receive the intervention. Data analysis will include the following descriptive statistics: correlations, ANOVA, and chi-square tests. Discussion The stepped wedge design may be appropriate for evaluating the effectiveness of a cancer health disparities program in a controlled setting. The stepped wedge design enables all designated correctional sites to eventually receive Cancer 101 during the study. We expect that Cancer 101 will prove to be a feasible program for reducing cancer health disparities in underserved men in correctional settings. Citation Format: Pamela Valera, Daniel Mackey, William McLaughlin, Laura Brotzman. Using a stepped wedge design to describe the adoption of a cancer education program in male correctional facilities. [abstract]. In: Proceedings of the Ninth AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2016 Sep 25-28; Fort Lauderdale, FL. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2017;26(2 Suppl):Abstract nr A39.

Abstract Inflammatory diseases such as ulcerative colitis recruit myeloid cells and result in inflammatory responses such as generation of reactive oxygen species (ROS). Development of non-invasive luminescence imaging approaches is needed to evaluate these inflammatory responses. We developed methodologies to show that vibration induced gut inflammation can be determined using luminescence imaging approach in a non-invasive fashion (Papineni and Vizard, World Molecular Imaging Congress 2012). Citrobacter rodentium (CR) is a natural bacterial pathogen that infects the distal colon of mice and induces transmissible murine colonic hyperplasia (TMCH). Here we determined the early inflammatory response (ROS activity) to intestinal infection by CR in a longitudinal study. We further assessed the effects of allelic variation on murine chromosome 11 (B6.CAST. 11) on the inflammatory response. The ROS activity was monitored in real-time in vivo using L-012 (8-amino-5-chloro-7-phenylpyridol [3,4-d]pyridazine-1,4(2H,3H) dione), a chemiluminescence reporter, using planar multimodal imaging system. 0.1 ml of 1 mg/ml L-012 probe was injected (i.p.) in control and the experimental mice that were subjected to CR infection. Necrosis was imaged (post-infection day-19) using a fluorescent probe containing a bis (zinc2+dipicolylamine, Zn- DPA) motif which binds with high affinity to apoptotic, necrotic, and bacterial surfaces. Significant difference in the basal gut ROS activity was found in mice with allelic variation on chromosome 11 compared to the wild type littermates. Robust elevation in ROS activity was observed in both the wild type and B6.CAST.11mice that were infected with CR (post infection 9, 12, 19 days). The variation in the levels of ROS activity however was very distinct between the wild type and B6.CAST.11 mice indicating the contribution of molecular determinants at the chromosome 11. The potential advantages in these real-time ROS monitoring methodologies by in vivo imaging, in understanding mechanisms of infection, inflammation, and development of better intervention strategies will be elaborated. Citation Format: Rao Papineni, Ishfaq Ahmed, Steven LeVine, Kenneth Bradley, William Mclaughlin, John Pizzonia, Shahid Umar. Molecular imaging of early inflammatory response during hyperplasia and/or colitis in response to bacterial infection. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2864. doi:10.1158/1538-7445.AM2013-2864

A near infrared (NIR) fluorescent polymeric nanoparticle, commercialized under the name X-Sight 761 (X761), was tested for compatibility with pre-clinical in vivo imaging applications. In one experiment, an optical clearance profile was obtained by performing whole animal fluorescence imaging over the course of 48 hours on mice injected intravenously with X761. In a second trial, a temporal biodistribution was assessed by conducting necropsy and ex vivo analysis of X761 tissue accumulation at selected time points over a 48 hours period after IV injection. Taken together, the data demonstrate a sustained distribution of X761 into all major tissues over the time course, with an extremely low net clearance from the animal. This unique behavior is attributed to cell uptake mediated by the polycationic surface of X761. These properties negate the use of X761 as a reporter within a classical targeted molecular probe construct, in which selective concentration at a target site and rapid clearance from background tissues are needed to develop contrast. Nevertheless, the brightness and stability of X761 is well suited for a range of other applications, ranging from broad based in vivo drug delivery to in vitro fluorescence assays.

Abstract Staphylococcus aureus (S. aureus) causes significant morbidity and mortality worldwide and a prime cause in hospital infections. Staphylococcus aureus bactereremia is prevalent in neutropenic cancer patients, and malignancies form a sizeable risk factor for methycillin-resistant S. aureus (MRSA) infections. Targeting the virulence products is a promising approach for developing novel therapeutics. A detailed understanding of the virulence factors and the resultant immune response is quite essential for such development. Here, we established an athymic mice model to assess the initial immune response to S.aureus inoculation. As the polymorphonuclear neutrophils(PMN)and macrophages are a part of the early response, determination of its myeloperoxidase (MPO) activity in vivo is established. MPO is an inflammatory heme protein and utilizes hydrogen peroxide in the process of reactive oxygen species generation. 24 hrs after the intra muscular injection of the S.aureus bacterial lysate, the MPO activity was detected by i.p injection of luminal in mice. The blue luminescence resulting from the MPO activity was imaged and the luminescence images were overlaid on planar X-ray images for anatomical co-registration. The results show a robust MPO activity in the mouse bladder. This increase in MPO activity as a result of neutrophil activation at the bladder region was further confirmed using non-invasive X-ray contrast imaging of the bladder and co-registering the luminescence and the X-ray density signals at the bladder. The initial response shown here has similarities with the mechanism suggested recently for myobacterium bovis bacillus Calmette-Guerin (BCG) treatment of bladder cancer. Bacterial components, its structures that are involved in eliciting the robust PMN infiltration in bladder will be very valuable in the developing better bladder cancer anti-tumor treatments. 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 4288. doi:1538-7445.AM2012-4288

Promotion of neutrophil‐mediated inflammation at the tumor enhances the anti‐tumor immune response. Here we investigated the possibilities of using nutriceuticals in induction of myeloperoxidase (MPO activity) in a tumor mouse model. We monitored in vivo MPO activity in a intraperitoneal ovarian cancer tumor mouse model by injecting (i.p) luminol. Multimodal imaging at different time points were performed to determine both the progression of the intraperitoneal tumor metastasis (fluorescence) and the MPO activity (lumiscence). The results show that basal MPO activity signal was not uniform between the tumors, and in some tumors, the MPO activity was completely absent indicating the presence of a differential tumor microenvironment within the mouse. Enhancement in MPO activity was observed upon administration of a single dose of 0.1 ml (50% volume/volume) of nutriceutical‐ BV9 (i.p injection). BV9 elicited a robust increase in the MPO activity within 3 hrs of its administration and the luminescence signals co localized with the tumors. Further investigations assessing the role of elicited MPO activity in cancer progression will greatly enhance the understanding of the tumor microenvironment and aid in better cancer treatments.

ProblemOvarian cancer stem cells (OCSCs) have been postulated as the potential source of recurrence and chemoresistance. Therefore identification of OvCSC and their complete removal is a pivotal stage for the treatment of ovarian cancer. The objective of the following study was to develop a new in vivo imaging model that allows for the detection and monitoring of OCSCs.Method of StudyOCSCs were labeled with X-Sight 761 Nanospheres and injected intraperitoneally (i. p.) and sub-cutaneously (s. c.) to Athymic nude mice. The Carestream In-Vivo Imaging System FX was used to obtain X-ray and, concurrently, near-infrared fluorescence images. Tumor images in the mouse were observed from different angles by automatic rotation of the mouse.Results X-Sight 761 Nanospheres labeled almost 100% of the cells. No difference on growth rate was observed between labeled and unlabeled cells. Tumors were observed and monitoring revealed strong signaling up to 21 days.Conclusion We describe the use of near-infrared nanoparticle probes for in vivo imaging of metastatic ovarian cancer models. Visualization of multiple sites around the animals was enhanced with the use of the Carestream Multimodal Animal Rotation System.

The microvasculature at the digits include the arteriovenous anastomoses (AVA) that enable fast increase or decrease of blood flow through the skin regulating body temperature. Determination of dermal blood flow changes at the phalanges and the inter‐phalangeal joints can be useful screening of atherosclerotic pathologies and endothelial dysfunction. A recent long‐term follow up study indicate that a positive response to cold pressor test is a potential predictor of hypertension. Sensitive methods to evaluate autonomic dysfunction and the related microcirculatory disturbances are warranted. Here, the autonomic sympathetic nervous system is assessed by measuring the cold induced pain related blood flow changes by Laser Doppler Flowmetry (LDF) and using a planar fluorescence imaging methods. Dermal blood flow was measured on the dominant hand while the non‐dominant hand of caucasian and asian adult volunteers were immersed in cold water up to 2 min. The resultant changes in the natural fluorescence were captured by the planar florescence imaging. The LDF measurements obtained before and after immersion show significant changes, with a 65% and above reduction in dermal blood flow from the baseline. The results indicate the possibility of utilizing imaging methodologies in microvascular dynamic analysis in clinical studies.

Abstract Molecular Imaging is making rapid strides as a tool in understanding and the cure of cancer. Like no panacea, here again a need of multimodality exists, where more than one modality is becoming a necessity to cogently decipher the role or action of the biomolecule in the pathology or cure. Here, we developed a multimodal nanoparticle (NP) that can be simultaneously be imaged by optical, and magnetic resonance Imaging (MRI) modalities. In combination, the two modalities compensate each other's deficiencies in the spatial resolution, sensitivity, and the depth detection. Encapsulation of MR contrast and amine functionalization was performed in a one pot synthesis by microemulsion technique with supra magnetic iron oxide (SPIO). The hydrodynamic radii of nanoparticle and zeta potential were measured using Nicomp 380 zls particle sizer. 9L gliosarcoma tumor bearing rat model was administered with the SPIO nanoparticles and validated using 7 tesla MRI scanner. PEGylated SPIO nanoparticles after conjugation with water soluble, near-infrared (NIR) tricarbocyanine, cyclic enamine-functionalized dye, were injected intratumorally and monitored noninvasively using near-IR fluorescence imager. Steady retention in the tumor as compared to the free near-infrared (NIR) dye injected into the control mice indicates the dye-colloidal particle dwell characteristics and suitability for targeted diagnostics. In summary, we have described the development of a simple and efficient dynamic Imaging multimodal nanoparticle to be used for NIRF and MR Imaging. 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 5329. doi:10.1158/1538-7445.AM2011-5329

Sessions C555Since CpxP has no homologues of known function, we have initially focused on its biophysical and structural characterization.Using multi-angle laser light scattering (MALLS), small-angle Xray scattering (SAXS) analysis, and formaldehyde-mediated crosslinking experiments, we show that full-length E. coli CpxP is a dimer in vivo as well as in pathway inactivating (pH 5.8) and activating (pH 8.0) conditions in vitro.Far-UV circular dichroism (CD) was used to demonstrate that CpxP is mainly α-helical, while near-UV CD and SAXS revealed that the protein may undergo a small structural adjustment in response to a pathway-inducing stimulus (pH 8.0).The crystal structure of CpxP, determined to 2.85 Å resolution, revealed an antiparallel dimer of intertwined α-helices with a highly basic concave surface.Each protomer consists of a long, hooked and bent hairpin fold with conserved LTXXQ motifs forming two diverging turns at one end.Three of six previously characterized cpxP loss-of-function mutations, M 59 T, Q 55 P, and Q 128 H, likely result from a destabilization of the protein fold, whereas the R 60 Q, D 61 E, and D 61 V mutations may alter interactions important for the signalling or proteolytic adaptor functions of CpxP.

A radiographic system is optimized for the contrast inherent to small animals and is developed for a multi-modal imaging system devised for in-vivo studies. The range of X-ray energies utilized (generally considered "soft X-rays") enables enhanced spatial resolution and superior contrast for detailed study of the mouse anatomy and smaller specimens. Despite the difficulties presented by the complicated energy spectrum of soft X-rays, relevant system calibrations for bone measures are described in detail and applied to the mouse. Further, long-bone symmetry modeling using a cylindrical projection is applied to the planar density image, providing convenient bone density estimates that are consistent with other methodologies.

IgG antibodies were conjugated to Kodak X-Sight nanospheres to develop fluorescent-labeled antibodies using two different synthetic routes: one involving the DTT reduction method, and the other involving Traut's Reagent modification method. These two methods result in different conjugation efficiencies and different performances in antigen detection. Western blotting shows that the nanosphere-IgG antibody conjugates synthesized using the DTT reduction method are more immunospecific than the conjugates synthesized using Traut's Reagent modification method. In addition, the conjugates synthesized using DTT reduction also show higher antigen detection sensitivity than other commercially available fluorescent-IgG antibody conjugates, including Alexa Fluor, Qdot, and CyDye conjugates.

23 million people in the United States alone.1 As such, scientists are actively researching ways to treat and slow the progression of such ailments. The primary means of studying the onset and progression of CKD and other renal disorders is to measure the glomerular filtration rate (GFR). Current methods of measuring kidney function involve the observation of creatinine and inulin clearance.1 Progression of CKD presents with a reduction in GFR, as measured by reduced creatinine and inulin clearance rates. Studies utilizing nonradioactive iodine X-ray contrast media have more recently been used in order to measure renal insufficiency.2,3 Iodinated contrast agents are a reliable measure of GFR through a bolus injection followed by blood sampling. This technique is used both in laboratory rats3 and in the clinic2 because the rate of contrast agent clearance can be measured reproducibly. Furthermore, administration of iodinated X-ray contrast media (iodixanol) produces no major changes in glomerular filtration rate and is a safe diagnostic tool in patients with kidney functions ranging from normal to end-stage renal disease.4,5