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Browsing by Author "Lipking, Kelsey"
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Item Effects of Thrombopoietin (TPO) on Longitudinal Mouse Hind Limb Crush Injury Model(Office of the Vice Chancellor for Research, 2014-04) Rothchild, Greg; Lipking, Kelsey; McKinley, Todd; Kacena, Melissa A.; Sandusky, George E.Approximately 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.Item Effects of Thrombopoietin (TPO) on Longitudinal Mouse Hind Limb Crush Injury Model(Office of the Vice Chancellor for Research, 2015-04-17) Rothchild, Greg; Lipking, Kelsey; McKinley, Todd; Kacena, Melissa A.; Sandusky, George E.Approximately 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 we utilized 9 separate mouse model 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. Crush models were introduced to hind limb crush injury by a mechanical-gravity driven Einhorn device. 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 Masson’s 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( less than day 3) , satellite cells, and neovascularization. A follow-up immunostain (CD206 specific for M2 double labeled with F4/80) was performed to characterize the macrophages in and around the lesions at day 3. M2 macrophages were seen around the periphery of the lesion and none in the middle of the lesion. There were very minimal differences in M2 numbers between the PBS and TPO treated groups at day 3. In conclusion, comparing the TPO treated mice versus the PBS control group with F4/80 immunostain showed the lesions at both time points were less in the TPO treated mice.Item ELEVATED LEVELS OF PLATELETS AND MDM2 EXPRESSION ARE CONTRIB-UTING FACTORS TO FACILITATING THE METASTASIS OF OSTEOSARCOMA(Office of the Vice Chancellor for Research, 2012-04-13) Lipking, Kelsey; Kacena, Melissa A.; Konopka, Jeff A.; Mayo, Lindsey D.; Sandusky, George E.Osteosarcoma (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.Item Increased Incidence of Lymphosarcoma in Long-Term Murine Survivors of Lethal Radiation: A Classification of Subtypes(Office of the Vice Chancellor for Research, 2013-04-05) Spencer, Cleandrea; Chua, Hui Lin; Plett, Arthur; Sampson, Carol; Joshi, Mandar; Roberts, Christopher S.; Lipking, Kelsey; Orschell, Christie M.; Sandusky, George E.Residual 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.Item Role of Complement Activation in Obliterative Bronchiolitis Post Lung Transplantation(The American Association of Immunologists, Inc., 2013-10-15) Suzuki, Hidemi; Lasbury, Mark E.; Fan, Lin; Vittal, Ragini; Mickler, Elizabeth A.; Benson, Heather L.; Shilling, Rebecca; Wu, Qiang; Weber, Daniel J.; Wagner, Sarah R.; Lasaro, Melissa; Devore, Denise; Wang, Yi; Sandusky, George E.; Lipking, Kelsey; Pandya, Pankita; Reynolds, John; Love, Robert; Wozniak, Thomas; Gu, Hongmei; Brown, Krista M.; Wilkes, David S.; Department of Medicine, School of Medicine,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.Item Use of Aperio Whole Slide Imaging System to Capture and Utilize Digital Virtual Slides for Pathology Education(Office of the Vice Chancellor for Research, 2014-04-11) Capouch, Samuel; Lipking, Kelsey; Surface, Ronne Leigh; Morgan, Randy; Sandusky, George E.Digital 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.