Use of Aperio Whole Slide Imaging System to Capture and Utilize Digital Virtual Slides for Pathology Education

poster abstractDigital whole slide imaging is the technique of digitizing an entire microscope slide at the highest resolution to produce a “digital virtual microscope slide” with high image quality. This digital image can be viewed in three to four fields, from low to high power, a feature commonly used by pathologists. This digital virtual slide can be used in conjunction with image processing software (both windows-based and browser-based) to view, manipulate, position, and specify the magnification of the image on a screen as if using a regular microscope to view the original glass slide. As the slide is captured in a virtual format, it is possible to use the image for archiving, copying, transferring over networks, distant consultation, as well as integration for educational use on the web and/or DVD. In this study, we captured all C603 and C604 sophomore pathology teaching slides in the general and systemic pathology course for viewing and learning through the Aperio ImageScope viewer. The resulting digital images possessed greater ease of use, were quicker to scan and allowed easier location of pathologic lesions in the slides. The ImageScope viewer allowed students to quickly zoom in and out of the slides at multiple fields of magnification. Instructors that have switched to the Aperio system from the old Bliss system found the Aperio system allowed the instructor to open up to 8 slides at one time, allowing side by side comparison to be completed on the same screen. The system also allows one to measure the size of the cells and to capture detailed images of tumor cells, inflammatory cells, and/or necrosis (cell death). This system is available for use on desktop, laptop, and most digital devices (such as smart phones or tablets). Compared to the old Bliss system, which is unable to perform these functions

Effects of Thrombopoietin (TPO) on Longitudinal Mouse Hind Limb Crush Injury Model

abstractApproximately 645 people suffer from blunt force trauma injury to the femur every day. The recovery time of such injury can last anywhere from 3-6 months. Thrombopoietin (TPO) was used as a growth factor to induce bone and muscle healing. In this study, nine separate mouse groups (10 mice per group) were used: Crush PBS, Crush TPO, Surgery PBS, and Surgery TPO at day 3 and day 17, and controls with no surgery/crush/treatment. Skeletal muscle was harvested from the following sites: experimental impact, experimental adjacent, and normal contralateral skeletal muscle as a control. The muscles were fixed, processed, sectioned, and stained with H&E and Massons Trichrome stains. The slides were reviewed for skeletal muscle injury, muscle necrosis, inflammation, muscle repair, and regeneration. In addition, F4/80, an immunostain for macrophages was performed. On microscopic examination at day 3 the most common histologic changes seen were sporadic muscle fiber vacuolation, focal necrosis of varying sizes, muscle contraction bands, and infiltration of macrophages. On day 17, the skeletal muscle injury was generally healed. The main histologic lesions seen were variable sizes of muscle fibers, early fibroplasia, fat infiltration, some macrophages, satellite cells, and neovascularization. Comparing the TPO treated mice versus the PBS control group, the lesions at both time points were less in the TPO treated mice

ELEVATED LEVELS OF PLATELETS AND MDM2 EXPRESSION ARE CONTRIB-UTING FACTORS TO FACILITATING THE METASTASIS OF OSTEOSARCOMA

poster abstractOsteosarcoma (OS) is the most common form of primary bone cancer and the 6th leading cause of cancer in pediatric patients. A chart review of OS patients treated at this institution suggests that a high platelet count at di-agnosis is significantly (p=0.023) and inversely associated with the first year of survival. As the effects of platelet interaction with OS have been exten-sively researched and suggest that platelets may facilitate tumor metastasis, and the most important prognostic factor for OS patient survival is metasta-sis to the lungs, we hypothesized that platelets increase metastasis to the lungs and reduce survival. Therefore, we sought to determine whether in-creasing platelet numbers in a well characterized OS mouse model would de-crease survival and/or increase metastasis to the lungs. We found that thrombopoietin (TPO) treated mice, had increased platelet numbers, died earlier than placebo treated controls, and that lungs from TPO treated mice contained a small number of large tumor cells (most metastatic lesions were 2-4 cells), whereas lungs from placebo treated controls showed no signs of metastases. Next, an OS tissue microarray (TMA) was built from OS patients seen at our institution over the past 10 years. Mdm2, p53, TPO, and c-mpl expression were evaluated by immunohistochemical (IHC) staining followed by quantitation using the Aperio Imaging system and analysis software. C-mpl (TPO receptor) expression was higher in the metastatic than the primary tumors, suggesting that platelets may contribute to the metastasis of OS. Elevated levels of Mdm2 correlated with metastasis and lower levels of p53, as detected by IHC. In conclusion, both the mouse model and the human OS data were similar, suggesting that both platelets and Mdm2 promote metas-tases in OS

Increased Incidence of Lymphosarcoma in Long-Term Murine Survivors of Lethal Radiation: A Classification of Subtypes

poster abstractResidual bone marrow damage (RBMD) persists for years following exposure to radiation and is thought to be due to decreased self-renewal of hematopoietic stem cells (HSC). We previously examined RBMD in murine survivors of lethal radiation modeling a terrorist event [800cGy total-body irradiation (TBI)]. We reported severely deficient HSC potential up to 20mo post-TBI compared to non-TBI age-matched controls, evidenced by minimal engraftment skewed to myeloid cells. CBC and BM cellularity were decreased in TBI mice, most dramatically in old age (>16mo). The percentage of some hematopoietic progenitors was consistently increased in TBI mice (~1.4x higher than non-TBI) possibly due to an increased cell cycling rate compared to non-TBI cells. Of interest, we now report the occurrence of a thymic mass developing in 13-24% of TBI mice 2-19 months post-TBI, compared to <1% of non-TBI. We characterized the Lymphosarcoma into the following groups based on the St. Jude pathology subclassification: Diffuse Lymphosarcoma involving multiple organs, Thymic lymphoma (usually associated with thymic and around the heart), Lymphosarcoma (potentially starting in the spleen and peri-pancreatic lymph nodes (Ab=abdomen)), and follicular lymphoma seen as a diffuse proliferation of lymphocytes in the white pulp area in the spleen. Thymic lymphomas were the most common, followed by Lymphosarcoma (Ab), follicular lymphoma (restricted to white pulp area in the spleen) and diffuse Lymphosarcoma. Immunostain markers revealed the thymic lymphomas were from T-cell lineage and the abdominal Lymphosarcoma were mainly from B-cell lineage. A few mice had disease involving the bone marrow. Taken together, these data suggest that the increased cycling among primitive hematopoietic cells in survivors of lethal radiation may contribute to stem cell exhaustion and subsequent RBMD, as well as predispose survivors to hematopoietic neoplasias

Oncology clinical trials and insurance coverage: An update in a tenuous insurance landscape

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/151873/1/cncr32360_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/151873/2/cncr32360.pd

"Allowing it to speak out of him": The Heterobiographies of David Malouf, Antonio Tabucchi and Marguerite Yourcenar

Under true pretences We know very little about the life of Ovid, and it is this absence of fact that has made him useful as the central figure of my narrative and allowed me the liberty of free invention, since what I wanted to write was neither historical novel nor biography, but a fiction with its roots in possible event. Thus starts the Afterword of David Malouf’s An Imaginary Life, a novel in which the poet Ovid, exiled from Rome, narrates his experience in the border outpost of Tomis, near the delta of the Danube on the Black Sea. ‘Relegated’ among the Getae at the edges of the Empire and ‘expelled form the confines of [the] Latin tongue’ (IL 26), this glittering and cynical poet undergoes a series of changes or metamorphoses. Initially pining for Rome and its sophisticated, complex language, he learns to overcome his hostility towards the barbarous people and their language, but when he discovers a wild Child that had been raised by the wolves in the forest and captures him with the intention of teaching him to speak and to be human, he soon realises that he himself has to learn from the Child another language, based not on symbolization and arbitrary convention but on an intuitive identity with things, on becoming the things signified in silence. After the death of the village’s elderly chief, which the villagers blame on the child’s demonic powers, the poet and the Child escape north across the frozen river. Ovid’s death is the poet’s final transformation, perhaps a literal metamorphosis like the ones described in Ovid’s great poem. Malouf’s Afterword concludes: My purpose was to make this glib fabulist of ‘the changes’ live out in reality what had been, in his previous existence, merely the occasion for dazzling literary display. (IL 154) Is Malouf’s novel then a fantasy inspired by ‘mere’ literary dazzle or, as ‘a fiction with its roots in possible event’, is it a work that, while not claiming to the factual accuracy of biography or the broad reliability of the historical background of a historical novel, can however still claim to be rooted in verisimilitude, in events that, although not documented, are nevertheless possible, as would be the case with a realist novel, or in Aristotelian poetics? The Afterword thematizes a tension between the desire to anchor the novel to history and the desire to free Ovid from historical necessity. How can Ovid live out ‘in reality’ the metamorphoses to which he is subjected, if metamorphoses are but the occasion for ‘literary display’? This tension also defines, more widely, the large number of novels written as if they were the autobiographies of historical personages, novels that gesture towards historical factuality and literary fictionality, towards ‘truth’ and invention, and exist under the sign of an essential structural displacement (the ‘autobiography’ is written by another) that brings to the foreground structural, narrative, and ethical issues also central to autobiography itself. (First paragraphs of submitted version

Apurinic/Apyrimidinic Endonuclease/Redox Factor-1 (APE1/Ref-1) redox function negatively regulates NRF2

Apurinic/apyrimidinic endonuclease/redox factor-1 (APE1/Ref-1) (henceforth referred to as Ref-1) is a multifunctional protein that in addition to its base excision DNA repair activity exerts redox control of multiple transcription factors, including nuclear factor κ-light chain enhancer of activated B cells (NF-κB), STAT3, activator protein-1 (AP-1), hypoxia-inducible factor-1 (HIF-1), and tumor protein 53 (p53). In recent years, Ref-1 has emerged as a promising therapeutic target in cancer, particularly in pancreatic ductal carcinoma. Although a significant amount of research has centered on Ref-1, no wide-ranging approach had been performed on the effects of Ref-1 inhibition and transcription factor activity perturbation. Starting with a broader approach, we identified a previously unsuspected effect on the nuclear factor erythroid-related factor 2 (NRF2), a critical regulator of cellular defenses against oxidative stress. Based on genetic and small molecule inhibitor-based methodologies, we demonstrated that repression of Ref-1 potently activates NRF2 and its downstream targets in a dose-dependent fashion, and that the redox, rather than the DNA repair function of Ref-1 is critical for this effect. Intriguingly, our results also indicate that this pathway does not involve reactive oxygen species. The link between Ref-1 and NRF2 appears to be present in all cells tested in vitro, noncancerous and cancerous, including patient-derived tumor samples. In particular, we focused on understanding the implications of the novel interaction between these two pathways in primary pancreatic ductal adenocarcinoma tumor cells and provide the first evidence that this mechanism has implications for overcoming the resistance against experimental drugs targeting Ref-1 activity, with clear translational implications

Hypoxia-Inducible Factor-1α Regulates CD55 in Airway Epithelium

Airway epithelial CD55 down-regulation occurs in several hypoxia-associated pulmonary diseases, but the mechanism is unknown. Using in vivo and in vitro assays of pharmacologic inhibition and gene silencing, the current study investigated the role of hypoxia-inducible factor (HIF)-1α in regulating airway epithelial CD55 expression. Hypoxia down-regulated CD55 expression on small-airway epithelial cells in vitro, and in murine lungs in vivo; the latter was associated with local complement activation. Treatment with pharmacologic inhibition or silencing of HIF-1α during hypoxia-recovered CD55 expression in small-airway epithelial cells. HIF-1α overexpression or blockade, in vitro or in vivo, down-regulated CD55 expression. Collectively, these data show a key role for HIF-1α in regulating the expression of CD55 on airway epithelium

Role of Complement Activation in Obliterative Bronchiolitis Post Lung Transplantation

Obliterative bronchiolitis (OB) post lung transplantation involves IL-17 regulated autoimmunity to type V collagen and alloimmunity, which could be enhanced by complement activation. However, the specific role of complement activation in lung allograft pathology, IL-17 production, and OB are unknown. The current study examines the role of complement activation in OB. Complement regulatory protein (CRP) (CD55, CD46, Crry/CD46) expression was down regulated in human and murine OB; and C3a, a marker of complement activation, was up regulated locally. IL-17 differentially suppressed Crry expression in airway epithelial cells in vitro. Neutralizing IL-17 recovered CRP expression in murine lung allografts and decreased local C3a production. Exogenous C3a enhanced IL-17 production from alloantigen or autoantigen (type V collagen) reactive lymphocytes. Systemically neutralizing C5 abrogated the development of OB, reduced acute rejection severity, lowered systemic and local levels of C3a and C5a, recovered CRP expression, and diminished systemic IL-17 and IL-6 levels. These data indicated that OB induction is in part complement dependent due to IL-17 mediated down regulation of CRPs on airway epithelium. C3a and IL-17 are part of a feed forward loop that may enhance CRP down regulation, suggesting that complement blockade could be a therapeutic strategy for OB

'Dressed in a Little Brief Authority': Authority Before, During, and After Shakespeare's Plays

This essay discusses the concept of authority from Shakespeare’s time to our own, considers examples of Shakespeare’s own explorations of authority, and provides a concise history of the ways in which Shakespeare has been constructed as cultural authority in different historical periods. It summarises the essays contained in the book, and argues that the book illuminates not only how Shakespeare became the archetypal figure of English cultural authority, but, perhaps more interestingly, why