bluuebunny/crossref_metadata_2025 huggingface dataset:
Plasma derived colostrum and life expectancy in calves
Authors: Dean Rachel , Wareham Kathryn
Published: October 2015

BestBETs for Vets are generated by the Centre for Evidence‐based Veterinary Medicine at the University of Nottingham to help answer specific questions and assist in clinical decision making. Although evidence is often limited, they aim to find, present and draw conclusions from the best available evidence, using a standardised framework. A more detailed description of how BestBETs for Vets are produced was published in Veterinary Record earlier this year (VR, April 4, 2015, vol 176, pp 354‐356)
Isotropic–kinematic hardening model for coarse granular soils capturing particle breakage and cyclic loading under triaxial stress space
Authors: Chen Qingsheng , Indraratna Buddhima , Carter John P. , Nimbalkar Sanjay
Published: October 2015

In this paper, a simple but comprehensive cyclic stress–strain model that incorporates particle breakage for granular soils including ballast and rockfill has been proposed on the basis of bounding surface plasticity theory within a critical state framework. Particle breakage and its effects are captured by a critical state line that is translated in voids ratio–stress space according to the dissipated energy (plastic work), through a hyperbolic function. A comprehensive equation related to particle breakage is proposed for the stress–dilatancy relationship to capture the complex dilatancy of granular soils. By extending Masing’s rule to bounding surface plasticity theory and introducing a generalized homological centre, a combined isotropic–kinematic hardening rule and a mapping rule have been established to simulate more realistically the response of gravelly soils under cyclic loading. The applicability and accuracy of this model are demonstrated by comparing its predictions with experimental results for different types of granular soils, including rockfill, under both monotonic and cyclic loading conditions. This study shows that the model can capture the characteristic features of coarse granular soils under complex loading paths.
Evaluation of an unmanned rotorcraft to monitor wintering waterbirds and coastal habitats in British Columbia, Canada
Authors: Drever Mark C. , Chabot Dominique , O'Hara Patrick D. , Thomas Jeffrey D. , Breault André , Millikin Rhonda L.
Published: October 2015

The effective protection of coastal and estuarine habitats requires reliable monitoring information on their use by waterbirds, and the use of small unmanned aircraft systems (UAS) may provide access to these habitats without disturbing birds. We evaluated the use of a rotary-wing UAS with a high-end consumer camera to identify and count wintering waterbirds at two coastal sites in British Columbia, Canada, in January 2015, and to map mudflat and marsh habitats. Photos of shorebirds, waterfowl, and seabird species were taken at varying altitudes, and disturbance of birds appeared minimal when the UAS was flown at heights ≥61 m. A ground resolution of ~1 cm/pixel was needed to discern plumage characteristics necessary to reliably identify birds. For some duck species, identification of females relied on body size or close association with a nearby male. Photographs were also used to derive accurate counts of shorebirds. For diving birds, accurate counts from photographs will require information on the proportion of birds on the water surface. Orthomosaics of coastal habitats were constructed with sufficient detail to assess ecological and geomorphological features. The UAS can therefore assist with bird species identification, population assessment, and characterization of habitat types.
Leedsichthys problematicus
: Arthur Smith Woodward's ‘most embarrassing enigma'
Authors: Liston J. J.
Published: October 2015

The link between the renowned palaeoichthyologist Arthur Smith Woodward and the similarly lauded marine reptile collector Alfred Nicholson Leeds may seem an unlikely one, but they formed a close family friendship during their professional acquaintance. Amongst the many fish specimens described by Smith Woodward from Leeds’ Oxford Clay collection, the large suspension feeder
Leedsichthys problematicus
was a challenge that he failed to resolve in print. Work is done to resolve the confused storage history of the material, in order to identify and reappraise the bones that comprised the type material, in the process revealing a key misidentification that undoubtedly coloured Smith Woodward's type description, and his initial understanding and interpretation of the animal.
Pesticide-Induced Stress in Arthropod Pests for Optimized Integrated Pest Management Programs
Authors: Guedes R.N.C. , Smagghe G. , Stark J.D. , Desneux N.
Published: October 2015

More than six decades after the onset of wide-scale commercial use of synthetic pesticides and more than fifty years after Rachel Carson's Silent Spring, pesticides, particularly insecticides, arguably remain the most influential pest management tool around the globe. Nevertheless, pesticide use is still a controversial issue and is at the regulatory forefront in most countries. The older generation of insecticide groups has been largely replaced by a plethora of novel molecules that exhibit improved human and environmental safety profiles. However, the use of such compounds is guided by their short-term efficacy; the indirect and subtler effects on their target species, namely arthropod pest species, have been neglected. Curiously, comprehensive risk assessments have increasingly explored effects on nontarget species, contrasting with the majority of efforts focused on the target arthropod pest species. The present review mitigates this shortcoming by hierarchically exploring within an ecotoxicology framework applied to integrated pest management the myriad effects of insecticide use on arthropod pest species.
A Uniform System for the Annotation of Vertebrate microRNA Genes and the Evolution of the Human microRNAome
Authors: Fromm Bastian , Billipp Tyler , Peck Liam E. , Johansen Morten , Tarver James E. , King Benjamin L. , Newcomb James M. , Sempere Lorenzo F. , Flatmark Kjersti , Hovig Eivind , Peterson Kevin J.
Published: October 2015

Although microRNAs (miRNAs) are among the most intensively studied molecules of the past 20 years, determining what is and what is not a miRNA has not been straightforward. Here, we present a uniform system for the annotation and nomenclature of miRNA genes. We show that less than a third of the 1,881 human miRBase entries, and only approximately 16% of the 7,095 metazoan miRBase entries, are robustly supported as miRNA genes. Furthermore, we show that the human repertoire of miRNAs has been shaped by periods of intense miRNA innovation and that mature gene products show a very different tempo and mode of sequence evolution than star products. We establish a new open access database—MirGeneDB ( http://mirgenedb.org )—to catalog this set of miRNAs, which complements the efforts of miRBase but differs from it by annotating the mature versus star products and by imposing an evolutionary hierarchy upon this curated and consistently named repertoire.
Experimental Work in Presupposition and Presupposition Projection
Authors: Schwarz Florian
Published: October 2015

Recent years have seen a surge of experimental approaches to the study of natural language meaning, both to obtain solid data on subtle phenomena that are hard to assess through introspection and to understand how abstract characterizations of linguistic knowledge relate to real-time cognitive processes in language comprehension. This article reviews research in one of the most recent areas to see extensive experimental investigations, namely presupposition and presupposition projection. Presuppositions are at the very nexus of linguistically encoded content and contextual information, as they relate directly to the discourse context but also interact in intricate ways with their intra-sentential linguistic environment. They are thus extremely suitable for investigations of the interplay of linguistic and more domain-general processes in language comprehension, as well as for experimental investigations of subtle theoretical phenomena.
Diagnosis and Therapy of Acute Myeloid Leukemia in the Era of Molecular Risk Stratification
Authors: Komanduri Krishna V. , Levine Ross L.
Published: October 2015

The diagnosis and risk stratification of acute myeloid leukemia (AML) primarily rely on morphologic analysis and assessment of karyotype by chromosome banding analysis. For decades, standard AML induction therapy has utilized the combination of anthracyclines and cytarabine. Despite the use of postremission therapy, less than half of patients with AML will be cured of their disease. Allogeneic hematopoietic stem cell transplantation combines cytoreductive chemotherapy with adoptive immunotherapy and may cure patients who fail chemotherapy alone. Recent advances in next-generation sequencing have yielded important insights into the molecular landscape of AML with normal karyotype. Integrated prognostic models incorporating somatic mutation analyses may outperform prediction based on conventional clinical and cytogenetic factors alone. We review the evolution of risk profiling of AML from the cytogenetic to molecular era and describe the implications for AML diagnosis and postremission therapy.
Imaging Glaucoma
Authors: Hood Donald C.
Published: October 2015

Optical coherence tomography (OCT) is changing the way glaucoma is studied and diagnosed. Glaucoma damages retinal ganglion cell (RGC) axons at the optic disc, and the resulting retrograde degeneration destroys the RGC bodies. OCT allows for a noninvasive measurement of both retinal nerve fiber (RNF) and RGC layer thickness. In this article, OCT techniques are described for studying the thinning of these layers due to glaucoma. We have learned that there is more damage to the macula (central ±8 deg) than previously thought, and a simple anatomical model provides an explanation for this finding. Further, OCT technology has led to improved understanding of the relationship between RGC and RNF layer loss and behavioral data. Finally, another imaging technique, adaptive optics, has allowed a better visualization and understanding of details that are often difficult or impossible to see with current OCT technology.
Plasma levels of microRNA in chronic kidney disease: patterns in acute and chronic exercise
Authors: Van Craenenbroeck Amaryllis H. , Ledeganck Kristien J. , Van Ackeren Katrijn , Jürgens Angelika , Hoymans Vicky Y. , Fransen Erik , Adams Volker , De Winter Benedicte Y. , Verpooten Gert A. , Vrints Christiaan J. , Couttenye Marie M. , Van Craenenbroeck Emeline M.
Published: October 2015

Exercise training is an effective way to improve exercise capacity in chronic kidney disease (CKD), but the underlying mechanisms are only partly understood. In healthy subjects (HS), microRNA (miRNA or miR) are dynamically regulated following exercise and have, therefore, been suggested as regulators of cardiovascular adaptation to exercise. However, these effects were not studied in CKD before. The effect of acute exercise (i.e., an acute exercise bout) was assessed in 32 patients with CKD and 12 age- and sex-matched HS ( study 1). miRNA expression in response to chronic exercise (i.e., a 3-mo exercise training program) was evaluated in 40 CKD patients ( study 2). In a subgroup of study 2, the acute-exercise induced effect was evaluated at baseline and at follow-up. Plasma levels of a preselected panel miRNA, involved in exercise adaptation processes such as angiogenesis (miR-126, miR-210), inflammation (miR-21, miR-146a), hypoxia/ischemia (miR-21, miR-210), and progenitor cells (miR-150), were quantified by RT-PCR. Additionally, seven miRNA involved in similar biological processes were quantified in the subgroup of study 2. Baseline, studied miRNA were comparable in CKD and HS. Following acute exercise, miR-150 levels increased in both CKD (fold change 2.12 ± 0.39, P = 0.002; and HS: fold change 2.41 ± 0.48 P = 0.018, P for interaction > 0.05). miR-146a acutely decreased in CKD (fold change 0.92 ± 0.13, P = 0.024), whereas it remained unchanged in HS. Levels of miR-21, miR-126, and miR-210 remained unaltered. Chronic exercise did not elicit a significant change in the studied miRNA levels. However, an acute exercise-induced decrease in miR-210 was observed in CKD patients, only after training (fold change 0.76 ± 0.15). The differential expression in circulating miRNA in response to acute and chronic exercise may point toward a physiological role in cardiovascular adaptation to exercise, also in CKD.
Effect of autonomic blocking agents on the respiratory-related oscillations of ventricular action potential duration in humans
Authors: van Duijvenboden Stefan , Hanson Ben , Child Nick , Orini Michele , Rinaldi Christopher A. , Gill Jaswinder S. , Taggart Peter
Published: October 2015

Ventricular action potential duration (APD) is an important component of many physiological functions including arrhythmogenesis. APD oscillations have recently been reported in humans at the respiratory frequency. This study investigates the contribution of the autonomic nervous system to these oscillations. In 10 patients undergoing treatment for supraventricular arrhythmias, activation recovery intervals (ARI; a conventional surrogate for APD) were measured from multiple left and right ventricular (RV) endocardial sites, together with femoral artery pressure. Respiration was voluntarily regulated and heart rate clamped by RV pacing. Sympathetic and parasympathetic blockade was achieved using intravenous metoprolol and atropine, respectively. Metroprolol reduced the rate of pressure development (maximal change in pressure over time): 1,271 (± 646) vs. 930 (± 433) mmHg/s; P < 0.01. Systolic blood pressure (SBP) showed a trend to decrease after metoprolol, 133 (± 21) vs. 128 (± 25) mmHg; P = 0.06, and atropine infusion, 122 (± 26) mmHg; P < 0.05. ARI and SBP exhibited significant cyclical variations ( P < 0.05) with respiration in all subjects with peak-to-peak amplitudes ranging between 0.7 and 17.0 mmHg and 1 and 16 ms, respectively. Infusion of metoprolol reduced the mean peak-to-peak amplitude [ARI, 6.2 (± 1.4) vs. 4.4 (± 1.0) ms, P = 0.008; SBP, 8.4 (± 1.6) vs. 6.2 (± 2.0) mmHg, P = 0.002]. The addition of atropine had no significant effect. ARI, SBP, and respiration showed significant coupling ( P < 0.05) at the breathing frequency in all subjects. Directed coherence from respiration to ARI was high and reduced after metoprolol infusion [0.70 (± 0.17) vs. 0.50 (± 0.23); P < 0.05]. These results suggest a role of respiration in modulating the electrophysiology of ventricular myocardium in humans, which is partly, but not totally, mediated by β-adrenergic mechanisms.
Intrauterine endotoxin-induced impairs pulmonary vascular function and right ventricular performance in infant rats and improvement with early vitamin D therapy
Authors: Mandell Erica , Powers Kyle N. , Harral Julie W. , Seedorf Gregory J. , Hunter Kendall S. , Abman Steven H. , Dodson R. Blair
Published: October 2015

High pulmonary vascular resistance (PVR), proximal pulmonary artery (PA) impedance, and right ventricular (RV) afterload due to remodeling contribute to the pathogenesis and severity of pulmonary hypertension (PH). Intra-amniotic exposure to endotoxin (ETX) causes sustained PH and high mortality in rat pups at birth, which are associated with impaired vascular growth and RV hypertrophy in survivors. Treatment of ETX-exposed pups with antenatal vitamin D (vit D) improves survival and lung growth, but the effects of ETX exposure on RV-PA coupling in the neonatal lung are unknown. We hypothesized that intrauterine ETX impairs RV-PA coupling through sustained abnormalities of PA stiffening and RV performance that are attenuated with vit D therapy. Fetal rats were exposed to intra-amniotic injections of ETX, ETX+vit D, or saline at 20 days gestation (term = 22 days). At postnatal day 14, pups had pressure-volume measurements of the RV and isolated proximal PA, respectively. Lung homogenates were assayed for extracellular matrix (ECM) composition by Western blot. We found that ETX lungs contain decreased α-elastin, lysyl oxidase, collagen I, and collagen III proteins ( P < 0.05) compared control and ETX+vit D lungs. ETX-exposed animals have increased RV mechanical stroke work ( P < 0.05 vs. control and ETX+vit D) and elastic potential energy ( P < 0.05 vs. control and ETX+vit D). Mechanical stiffness and ECM remodeling are increased in the PA ( P < 0.05 vs. control and ETX+vit D). We conclude that intrauterine exposure of fetal rats to ETX during late gestation causes persistent impairment of RV-PA coupling throughout infancy that can be prevented with early vit D treatment.
Effects of hindlimb unloading and ionizing radiation on skeletal muscle resistance artery vasodilation and its relation to cancellous bone in mice
Authors: Prisby Rhonda D. , Alwood Joshua S. , Behnke Brad J. , Stabley John N. , McCullough Danielle J. , Ghosh Payal , Globus Ruth K. , Delp Michael D.
Published: October 2015

Spaceflight has profound effects on vascular function as a result of weightlessness that may be further compounded by radiation exposure. The purpose of the present study was to assess the individual and combined effects of hindlimb unloading (HU) and radiation (Rad) on vasodilator responses in the skeletal muscle vasculature. Adult male C57BL/6J mice were randomized to one of four groups: control (Con), HU (tail suspension for 15 days), Rad (200 cGy of137Cs), and HU-Rad (15-day tail suspension and 200 cGy of137Cs). Endothelium-dependent vasodilation of gastrocnemius feed arteries was assessed in vitro using acetylcholine (ACh, 10−9–10−4M) and inhibitors of nitric oxide synthase (NOS) and cyclooxygenase (COX). Endothelium-independent vasodilation was assessed using Dea-NONOate (10−9–10−4M). Endothelium-dependent and -independent vasodilator responses were impaired relative to Con responses in all treatment groups; however, there was no further impairment from the combination of treatments (HU-Rad) relative to that in the HU and Rad groups. The NOS-mediated contribution to endothelium-dependent vasodilation was depressed with HU and Rad. This impairment in NOS signaling may have been partially compensated for by an enhancement of PGI2-mediated dilation. Changes in endothelium-dependent vasodilation were also associated with decrements in trabecular bone volume in the proximal tibia metaphysis. These data demonstrate that the simulated space environment (i.e., radiation exposure and unloading of muscle and bone) significantly impairs skeletal muscle artery vasodilation, mediated through endothelium-dependent reductions in NOS signaling and decrements in vascular smooth muscle cell responsiveness to NO.
Physical activity-induced remodeling of vasculature in skeletal muscle: role in treatment of type 2 diabetes
Authors: Laughlin M. Harold
Published: October 2015

This manuscript summarizes and discusses adaptations of skeletal muscle vasculature induced by physical activity and applies this understanding to benefits of exercise in prevention and treatment of type 2 diabetes (T2D). Arteriolar trees of skeletal muscle are heterogeneous. Exercise training increases capillary exchange and blood flow capacities. The distribution of vascular adaptation to different types of exercise training are influenced by muscle fiber type composition and fiber recruitment patterns that produce different modes of exercise. Thus training-induced adaptations in vascular structure and vascular control in skeletal muscle are not homogeneously distributed throughout skeletal muscle or along the arteriolar tree within a muscle. Results summarized indicate that similar principles apply to vascular adaptation in skeletal muscle in T2D. It is concluded that exercise training-induced changes in vascular gene expression differ along the arteriolar tree and by skeletal muscle fiber type composition. Results suggest that it is unlikely that hemodynamic forces are the only exercise-induced signals mediating the regulation of vascular gene expression. In patients with T2D, exercise training is perhaps the most effective treatment of the many related symptoms. Training-induced changes in the vasculature and in insulin signaling in the muscle fibers and vasculature augment glucose and insulin delivery as well as glucose uptake. If these adaptations occur in a sufficient amount of muscle mass, exposure to hyperglycemia and hyperinsulinemia will decrease along with the risk of microvascular complications throughout the body. It is postulated that exercise sessions in programs of sufficient duration, that engage as much skeletal muscle mass as possible, and that recruit as many muscle fibers within each muscle as possible will produce the greatest benefit. The added benefit of combined resistance and aerobic training programs and of high-intensity exercise programs is not simply “more exercise is better”.
Abstract A12: PDL-1 blockade and Sunitinib enhance the efficiency of oncolytic viral therapy
Authors: Mostafa Ahmed , Gratton Kathy , Lawson Keith A. , Shi Zhong-Qiao , Thirukkumaran Chandini , Morris Don G.
Published: October 2015

Purpose: Oncolytic viruses (OV) are attractive for use against multiple tumours given their ability to prime therapeutic immune responses in addition to their direct cytotoxic effects. We investigated the utility of reovirus, an OV currently in phase III clinical trials, as a novel immunotherapeutic against renal cell (RCC), non small cell lung (NSCLC) and breast carcinomas (BrCa) as both a monotherapy and in combination with sunitinib, a first line metastatic RCC agent. In addition the utility of adding an immune checkpoint inhibitor was investigated.

Experimental Design: In-vitro, cytotoxicity, viral replication and chemokine production were assessed in a panel of human and murine cancer cell lines following exposure to reovirus, sunitinib or their combination. In-vivo, RENCA (murine renal cell carcinoma) cells were implanted subcutaneously into female balb/c mice to establish a syngeneic immunocompetent model of RCC. Tumor growth and overall survival were assessed following treatment with reovirus, sunitinib or their combination. IFN-γ, myeloid-derived suppressor cells (MDSC), and protective immunity were assessed by ELISA, flow cytometry (Gr1+/CD11b+) and adoptive transfer experiments, respectively. PDL-1 and PD-1 expression on both tumour and immune cells were assayed.

Results: In vitro, exposure of RCC cell lines to reovirus resulted in an inflammatory cytotoxic response that was augmented through combination with sunitinib. In vivo, reovirus significantly reduced RENCA tumor burden and generated an anti-tumor immune response that was augmented by sunitinib. Combinations with sunitinib also lead to a decrease in myeloid-derived suppressor cells (MDSC) and resulted in improved overall survival and enhanced protective immunity. Preliminary results in NSCLC and BrCa are suggestive that this strategy maybe applicable to many cancer histologies. Both PDL-1 and PD-1 expression were up regulated by RV (similar to IFN gamma) in the majority of cell lines tested suggesting a potential role for checkpoint blockade (CB).

Conclusions: Reovirus has both direct oncolytic and immunotherapeutic activity against RCC, which is augmented through combination with sunitinib. The role of immune CB is ongoing. These proof-of-principle investigations provide clear rationale to investigate RV/sunitinib/immune CB as a promising approach for early phase clinical trials as a novel treatment strategy for cancer.

Citation Format: Ahmed Mostafa, Kathy Gratton, Keith A. Lawson, Zhong-Qiao Shi, Chandini Thirukkumaran, Don G. Morris. PDL-1 blockade and Sunitinib enhance the efficiency of oncolytic viral therapy. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A12.
Abstract A18: A systems biology approach to predict immunotherapy augmented abscopal effects
Authors: Poleszczuk Jan , Prokopiou Sotiris , Robertson-Tessi Mark , Luddy Kimberly , Moros Eduardo , Fishman Mayer , Djeu Julie , Enderling Heiko
Published: October 2015

Introduction and Objectives: Radiation-induced abscopal effects are characterized by regression of tumor nodules outside the local radiation treatment field. Abscopal effects have been reported in metastatic lymphoma, melanoma, lung and breast cancer as well as renal cell carcinoma amongst others. Radiotherapy causes a perturbation to the local immune profile that is propagated throughout the lymphatic system, shifting systemic immune surveillance towards tumor eradication in distant, non-irradiated sites as well as circulating micrometastases. We hypothesize that abscopal effects could be triggered by targeting combined radiation and immunotherapy to immunogenic hubs in the patient-specific interconnected metastatic-lymphatic network.

Materials and Methods: We develop a mathematical model of the systemic response to local radio-immunotherapy. Patient-specific positron emission tomography (PET) scans provide the size and location of each tumor nodule. Metastatic geography is mapped onto the lymphatic network and connections between metastatic sites and lymphatic nodes are calculated. We quantify immune signal flow through the lymphatic network and amplifications through metastatic sites. A local model describes the temporal coevolution of tumor size with effector and regulatory T cells. The local and global models are coupled to quantify systemic responses to local treatment of each tumor site.

Results: The ‘virtual patient’ framework successfully simulates the abscopal effect reported for 2 patients with widely metastatic melanoma and non-small cell lung cancer. Model simulations suggest that only one or two tumor nodules in each patient serve as immunogenic hubs that could trigger an abscopal response. Radio-immunotherapy of any other lesion would result in local tumor control but systemic disease progression.

Conclusions: Careful selection of immunogenic ‘hub’ nodules holds the key to successful abscopal disease control. The presented mathematical framework may help to identify the immunogenicity of each tumor nodule. Before serving as a novel decision support tool, rigorous calibration and validation remains to be pursued as more clinical abscopal data becomes available.

Citation Format: Jan Poleszczuk, Sotiris Prokopiou, Mark Robertson-Tessi, Kimberly Luddy, Eduardo Moros, Mayer Fishman, Julie Djeu, Heiko Enderling. A systems biology approach to predict immunotherapy augmented abscopal effects. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A18.
Abstract A59: Novel inhibitors of human IDO1 and TDO
Authors: Banerjee Monali , Middya Sandip , Shrivastava Ritesh , Raina Sushil , Yadav Dharmendra , Singh Satveer , Manna Anindita , Chakraborty Debjani , Surya Arjun
Published: October 2015

In several human cancers, increased production of the tryptophan metabolite kynurenine by the enzymes indoleamine 2,3-dioxygenase (IDO1) and tryptophan 2,3-dioxygenase (TDO) creates a tolerogenic environment that promotes tumor driven immune suppression. One or both of these enzymes is up-regulated in a number of immune cells and in approximately 50% of human tumors (Pilotte et al., 2012). We have designed and synthesized novel inhibitors of IDO1 and dual inhibitors of IDO1 and TDO (PCT/US14/24920) that inhibit IDO1 in vitro and drop kynurenine production in established human cellular models of IDO1 (IFNγ treated HeLa cells) and/or TDO (A172 glioblastoma cells) and enhance T cell proliferation in a mixed culture assay. Further, these molecules cause a steep and durable reduction in plasma kynurenine in LPS treated c57/Bl6 mice, demonstrating inverse correlation between drug and kynurenine levels. In order to evaluate their therapeutic potential we monitored tumor penetration by the compounds and concomitant tumor kynurenine reduction in Balb/c mice implanted with the murine colon carcinoma CT-26 cell line. Several compounds drastically reduced tumor kynurenine levels compared to vehicle treated animals. One such compound, Compound 1, was further evaluated for its ability to reduce tumor burden both as monotherapy and in combination with doxorubicin. Currently we have nominated candidate compounds which are either IDO1 specific or IDO1/TDO dual inhibitors for regulatory toxicology.

Citation Format: Monali Banerjee, Sandip Middya, Ritesh Shrivastava, Sushil Raina, Dharmendra Yadav, Satveer Singh, Anindita Manna, Debjani Chakraborty, Arjun Surya. Novel inhibitors of human IDO1 and TDO. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A59.
Abstract A80: Validation and clinical correlation of triplex CD3, CD8 and FOXP3 IHC of tumor-infiltrating lymphocytes in follicular lymphoma
Authors: Mansfield James R. , Slater Christian , Byers Richard
Published: October 2015

It is clear that immune cells play many sometimes conflicting roles in the tumor microenvironment and it would be extremely useful to be able to visualize the distributions of multiple phenotyped immune cells in-situ in solid tumors. However, obtaining phenotypic information about the various cells that play these roles in and around the tumor has been a challenge. Existing methods can either deliver phenotypic information on homogenous samples (e.g., flow cytometry) or morphologic information on single immunomarkers (standard IHC). These limitations can be largely overcome through a multiplexed staining, imaging and analysis methodology using standard clinical FFPE tissue sections. Although multiplex methods have been shown to be particularly useful not much has been presented on the validation of such methods. We present here a validation of the method for CD3, CD8 and FOXP3 in tissue microarray containing triplex follicular lymphoma cores from 40 subjects [24 male, 16 female, age 35 to 75 years at diagnosis, median 55 years , 2- 171 months follow-up]. This involves a sequential multi-marker labeling for 3 antigens and a counterstain; automated multispectral imaging to separate chromogens; and an automated analysis that can quantitate the per-cell marker expression, determine the cellular phenotype, count these cells separately in the tumor compartment and in the stroma and provide high-resolution images of their distributions. The IHC for each marker was optimized in as single-stain IHC and then those specifications used as a part of the triplex stain. The samples were scored using an automated scoring methodology and the results from the triplexed method are compared to analyses done on singly stained sections with excellent correlation (R greater than 0.9 in all cases), showing that multiplexed staining methods can replicate standard IHC methods while maintaining the inter-distribution and visualization of the markers in a single section.

Citation Format: James R. Mansfield, Christian Slater, Richard Byers. Validation and clinical correlation of triplex CD3, CD8 and FOXP3 IHC of tumor-infiltrating lymphocytes in follicular lymphoma. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A80.
Abstract A82: The cholesterol metabolite, 27-hydroxycholesterol, promotes breast cancer metastasis
Authors: Nelson Erik Russell , Wardell Suzanne E. , McDonnell Donald P.
Published: October 2015

Breast cancer remains the most common cancer in women and is the second leading cause of cancer death in women. It is important to note that the majority of patients eventually succumb to the metastatic spread of their disease, not to the primary tumor itself. The magnitude of this problem provides strong rationale for studies that may lead to the development of new chemopreventative strategies and/or lifestyle changes that reduce breast cancer metastasis. It is of significance, therefore, that patients taking inhibitors of 2-hydroxy-3-methylglutaryl coenzyme A (statins) experience a significantly increased relapse free survival time. Therefore, our recent finding that 27-Hydroxycholesterol (27HC), a primary metabolite of cholesterol, exerts partial agonist activity on both the estrogen receptors (ERs) and liver X receptors (LXRs), suggests a potential mechanistic link between hypercholesterolemia and breast cancer pathology. We demonstrate that 27HC (1) increases in vivo tumor growth and lung metastases, (2) promotes an epithelial to mesenchymal like transition, and (3) in a myeloid cell dependent manner, alters the metastatic microenvironment to promote cancer cell colonization. Our ongoing work is aimed at identifying the specific mechanisms by which 27HC induces these changes and thereby increases metastasis. In summary, our data thus far strongly indicate a role for 27HC in breast cancer pathophysiology and metastasis. These results provide additional support for the exploration of potential chemopreventative benefits of lower cholesterol diets, pharmacological inhibitors of HMG-CoA reductase or the enzyme responsible for the synthesis of 27HC (CYP27A1), and macrophage ablative strategies.

This work was supported by an NIH-NCI grant to ERN (4R00CA172357-03).

Citation Format: Erik Russell Nelson, Suzanne E. Wardell, Donald P. McDonnell. The cholesterol metabolite, 27-hydroxycholesterol, promotes breast cancer metastasis. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A82.
Abstract A83: The role of Ikaros in effector and regulatory T cell homeostasis in a murine pancreatic cancer model
Authors: Nelson Nadine , Xiang Shengyan , Zhang Xiaohong , Szekeres Karoly , Tomar Ghansah
Published: October 2015

The alternatively spliced transcription factor Ikaros is crucial for normal lymphocyte development. Ikaros plays a role as a tumor suppressor in murine T cells. In pancreatic cancer hosts, the balance between effector and regulatory T cells is lost, leading to immunosuppression and reduced anti-tumor immunity. In this study, we aim to identify the role of Ikaros in regulating effector CD4+ and CD8+ and regulatory T cell homeostasis in a pancreatic tumor microenvironment. Using our murine heterotopic model of pancreatic cancer, we isolated splenocytes from tumor-bearing (TB) and control mice and performed western blot and qRT-PCR analyses to evaluate Ikaros mRNA and protein expression, respectively. Flow cytometry analyses were also used to immunophenotype T cell populations in splenocytes from control and TB mice. Next, magnetic activated cell sorting (MACS) was used to sort CD3+ T cells from TB and control splenocytes for western blot analyses of Ikaros expression. We also carried out in vitro and in vivo experiments using apigenin, a plant flavonoid and protein kinase inhibitor with anti-inflammatory, anti-tumor and anti-proliferative properties, to shed insight into how Ikaros may be regulated and its involvement in T cell development. Our results showed no difference in Ikaros mRNA expression but reduced expression of all Ikaros isoforms in splenocytes from TB mice compared with controls. Ikaros protein expression was also downregulated in sorted TB T cells, compared with control. Analyses of T cells by flow cytometry revealed a significant decrease in effector CD4+ and CD8+ T cell percentages but an increase in CD4+CD25+ regulatory T cells (Treg) in TB splenocytes compared with controls. In vitro, treatment of splenocytes with apigenin stabilized Ikaros' protein expression. In the presence of Panc02 cells, Ikaros protein expression was downregulated. However, the addition of apigenin to the co-culture blocked this downregulation. These correlated with in vivo findings that showed partial reversal of some Ikaros isoforms in TB mice treated with apigenin. Apigenin treatment also significantly increased CD4+ and CD8+ T cell percentages but decreased Treg percentages. We are currently performing experiments to determine how Ikaros may be regulated in T cells and its effect on their functions. Thus far, our findings suggest that PC progression is associated with reduced Ikaros expression, which may lead to loss of T cell homeostasis and thus, dampened anti-tumor immune responses.

Citation Format: Nadine Nelson, Shengyan Xiang, Xiaohong Zhang, Karoly Szekeres, Ghansah Tomar. The role of Ikaros in effector and regulatory T cell homeostasis in a murine pancreatic cancer model. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A83.
Abstract A84: IRF8 regulates GM-CSF expression in T cells and tumor cells to mediate myeloid-derived suppressor cell differentiation
Authors: Paschall Amy , Zhang Ruihua , Bardhan Kankana , Qi Chen-Feng , Peng Liang , Lu Geming , Yang Jianjun , Merad Miriam , Zimmerman Mary , McGaha Tracy , Zhou Gang , Mellor Andrew , Abrams Scott I. , Morse Herbert , Ozato Keiko , Xiong Huabao , Liu Kebin
Published: October 2015

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR05) of the Conference Proceedings.

Citation Format: Amy Paschall, Ruihua Zhang, Kankana Bardhan, Chen-Feng Qi, Liang Peng, Geming Lu, Jianjun Yang, Miriam Merad, Mary Zimmerman, Tracy McGaha, Gang Zhou, Andrew Mellor, Scott I. Abrams, Herbert Morse, Keiko Ozato, Huabao Xiong, Kebin Liu. IRF8 regulates GM-CSF expression in T cells and tumor cells to mediate myeloid-derived suppressor cell differentiation. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr A84.
Abstract B19: Targeting histone deacetylase 6 in T-cells to improve melanoma immunotherapy
Authors: Laino Andressa Sodre , Woods David M. , Celis Esteban , Weber Jeffrey , Sotomayor Eduardo
Published: October 2015

This abstract is being presented as a short talk in the scientific program. A full abstract is printed in the Proffered Abstracts section (PR09) of the Conference Proceedings.

Citation Format: Andressa Sodre Laino, David M. Woods, Esteban Celis, Jeffrey Weber, Eduardo Sotomayor. Targeting histone deacetylase 6 in T-cells to improve melanoma immunotherapy. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B19.
Abstract B51: Treatment of melanoma with a serotype 5/3 chimeric oncolytic adenovirus coding for GM-CSF: Results in vitro, in rodents and in humans
Authors: Bramante Simona , Kaufmann Johanna K. , Veckman Ville , Nettelbeck Dirk M. , Hemminki Otto , Liikanen Ilkka , Vassilev Lotta , Cerullo Vincenzo , Oksanen Minna , Heiskanen Raita , Joensuu Timo , Kanerva Anna , Pesonen Sari , Vähä-Koskela Markus , Koski Anniina , Hemminki Akseli
Published: October 2015

Metastatic melanoma is refractory to irradiation and chemotherapy, but amenable to immunological approaches such as ipilimumab or adoptive cell therapies. Oncolytic virus replication is an immunogenic phenomenon, and viruses can be armed with immunostimulatory molecules. Therefore, oncolytic immuno-virotherapy of malignant melanoma is an appealing approach, which was recently validated by a positive phase 3 trial.

We investigated the potency of oncolytic adenovirus Ad5/3-D24-GMCSF on a panel of melanoma cell lines and animal models in combination with low-dose cyclophosphamide, and summarized the melanoma-specific human data from the Advanced Therapy Access Program (ATAP).

The virus effectively eradicated human melanoma cells in vitro and subcutaneous SK-MEL-28 melanoma xenografts in nude mice when combined with low-dose cyclophosphamide. Furthermore, virally-expressed granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulated the differentiation of human monocytes into macrophages. In contrast to human cells, RPMI 1846 hamster melanoma cells exhibited no response to oncolytic viruses and the chimeric 5/3 fiber failed to increase the efficacy of transduction, suggesting limited utility of the hamster model in the context of viruses with this capsid. Four out of nine melanoma patients treated in ATAP were evaluable for possible therapy benefit with modified Response Evaluation Criteria In Solid Tumors (RECIST): one patient had minor response, two had stable disease, and one had progressive disease. Treatments appeared safe and well-tolerated. Two patients were alive at 559 and 1501 days after treatment.

Ad5/3-D24-GMCSF showed promising efficacy in preclinical studies and possible antitumor activity in melanoma patients refractory to other forms of therapy. This data supports continuing the clinical development of oncolytic adenoviruses for treatment of malignant melanoma.

Citation Format: Simona Bramante, Johanna K. Kaufmann, Ville Veckman, Dirk M. Nettelbeck, Otto Hemminki, Ilkka Liikanen, Lotta Vassilev, Vincenzo Cerullo, Minna Oksanen, Raita Heiskanen, Timo Joensuu, Anna Kanerva, Sari Pesonen, Markus Vähä-Koskela, Anniina Koski, Akseli Hemminki. Treatment of melanoma with a serotype 5/3 chimeric oncolytic adenovirus coding for GM-CSF: Results in vitro, in rodents and in humans. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B51.
Abstract B66: Tumor-specific differences in virus-induced inflammation may determine therapeutic response to HSV virotherapy in sarcoma models
Authors: Chen Chun-Yu , Hutzen Brian J. , Wang Pin-Yi , Leddon Jennifer L. , Currier Mark A. , Denton Nicholas , Eubank Timothy , Cripe Timothy P.
Published: October 2015

Ewing's Sarcoma (EWS) is the second most common childhood bone cancer with high rate of metastasis (25%) and tumor recurrence (30~40%). Despite tremendous advances in the five-year survival rate of localized disease (70~80%), overall survival for patients with relapse or metastasis remains poor (<30%). Oncolytic virotherapy is an emerging anticancer treatment that utilizes attenuated viruses to selectively target and destroy tumor cells. Promotion of systemic anti-tumor immune responses by virotherapy holds great promise, as it has the potential to target distant metastasis and provide immunologic protection from tumor relapse. We tested the oncolytic herpes simplex virus (oHSV) rRp450 as a novel therapy for sarcomas using two human EWS cell lines 5838 and A673 in vitro and in mouse xenografts. Interestingly, our data showed a significant disconnect between the susceptibility of the EWS cell lines 5838 and A673 grown in vitro and the virus-induced antitumor efficacy in vivo. Although 5838 displayed much less virus replication and substantially weaker susceptibility to rRp450 in vitro, it displayed a superior in vivo therapeutic response to intratumoral rRp450 injection compared to tumors derived from the more susceptible A673 cell line. Analysis of these two tumor types demonstrated significant differences on baseline and virus-induced tumor microenvironment characteristics, including differences in cytokines, chemokines, and immune cell infiltrates. In the 5838 model, rRp450 treatment triggered higher inflammatory immune responses, including inductions of interferon-gamma (IFN-γ), Natural Killer (NK) cell chemokine CXCL10 and increased infiltration of NK cells and neutrophils in tumors. In addition, the number of pro-tumorigenic CD206+ M2-type macrophages also significantly decreased in rRp450-treated 5838 tumors. Blockade of NK cells or neutrophils alone had no effect on oHSV-induced antitumor efficacy, suggesting neither of these cell populations alone is crucial for virus-induced antitumor efficacy. Efforts to identify key mediator(s) to oHSV-induced antitumor efficacy are currently under investigation. Taken together, our study suggests that, rather than the persistence of virus replication, the oHSV-induced immune inflammatory response might determine the outcome of the oncolytic virotherapy in these models.

Citation Format: Chun-Yu Chen, Brian J. Hutzen, Pin-Yi Wang, Jennifer L. Leddon, Mark A. Currier, Nicholas Denton, Timothy Eubank, Timothy P. Cripe. Tumor-specific differences in virus-induced inflammation may determine therapeutic response to HSV virotherapy in sarcoma models. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B66.
Abstract B73: Ketogenic diet enhances immunity to glioblastoma
Authors: Johnson John L. , Woolf Eric , Lussier Danielle M. , Brooks Ken S. , Blattman Joseph N. , Scheck Adrienne C.
Published: October 2015

Glioblastomas are the most common and malignant brain tumor in adults and account for over 50% of all gliomas. Current standard of care combines surgical resection with temozolomide chemotherapy and radiotherapy, but 5 year survival rates remain less than 20%. There is a critical need for new therapies that increase efficacy without increasing toxicity. One therapeutic target can be found in the aberrant metabolism that is a hallmark of cancer. We have previously used an intracranial syngeneic immunocompetent mouse model of malignant glioma to demonstrate that targeting metabolism using a ketogenic diet reduced tumor burden and increased survival. The ketogenic diet is a high fat, low carbohydrate plus protein diet used for the treatment of refractory pediatric epilepsy. Mice maintained on a ketogenic diet (KetoCal®, Nutricia North America, Gaithersburg, MD) had a reduction in inflammation and peri-tumoral edema, prompting us to question the effects of the ketogenic diet on the infiltration and the functionality of glioblastoma-reactive immune cells. Immune cells were isolated from malignant gliomas from mice treated with either the ketogenic diet or a standard diet. Glioblastoma-specific immune cells were tested for functionality via cytokine production, capacity for cytotoxicity, and amount of infiltration. We found the ketogenic diet significantly decreased expression of the inhibitory receptors CTLA4 and PD-1 on tumor-infiltrating T cells, and coincided with a reduction in inhibitory ligand expression on the tumor. Moreover, this reduced T cell tolerance at the tumor site allowed CD8+ T cells to produce IFNγ and IL-2, and regain cytotoxic functions. Additionally, the ketogenic diet caused a reduction in immunosuppressive cytokine production via regulatory T cells. Innate immune responses to the tumor were also altered in mice maintained on the ketogenic diet, as natural killer cells infiltrating the tumor were able to produce more IFNγ and TNF in response to GL261-luc2 cells in vitro. Overall, the ketogenic diet may be an attractive adjuvant therapy to overcome several immune escape mechanisms in gliomas by decreasing regulatory T cell suppression of effector T cells and increasing CD8+ T cell killing and cytokine production, ultimately leading to increased tumor immune mediated rejection.

Citation Format: John L. Johnson, Eric Woolf, Danielle M. Lussier, Ken S. Brooks, Joseph N. Blattman, Adrienne C. Scheck. Ketogenic diet enhances immunity to glioblastoma. [abstract]. In: Proceedings of the AACR Special Conference: Tumor Immunology and Immunotherapy: A New Chapter; December 1-4, 2014; Orlando, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2015;3(10 Suppl):Abstract nr B73.