Podocalyxin in the Diagnosis and Treatment of Cancer

Kelly M. McNagny, Michael R. Hughes, Marcia L. Graves, Erin J. DeBruin, Kimberly Snyder, Jane Cipollone, Michelle Turvey, Poh C. Tan, Shaun McColl and Calvin D. Roskelle

Copernicus Ocean State Report, issue 6

The 6th issue of the Copernicus OSR incorporates a large range of topics for the blue, white and green ocean for all European regional seas, and the global ocean over 1993–2020 with a special focus on 2020

The apical membrane protein podocalyxin acts as an anti-adhesin in epithelial ovarian carcinoma

Podocalyxin is a heavily sialylated and sulfated integral membrane protein with anti-adhesive properties that contributes to apical domain formation in normal, polarized epithelial cells. Immunohistochemical analysis of an outcome-linked ovarian tumour microarray (TMA) revealed that podocalyxin was expressed in the majority of lesions but showed no significant correlation with poor outcome. Podocalyxin expression was most prevalent in serous tumours where it was often apically localized in low-grade well-differentiated tumours. In the normal ovary, podocalyxin expression was moderate and localized to the free peritoneal surface of normal ovarian surface epithelia (OSE), serving to distinguish the OSE from its underlying stroma. Interestingly, podocalyxin was apically localized in dysplastic, columnar OSE suggesting that it may serve as a marker of polarity early in the carcinogenic process. After characterizing podocalyxin expression in a variety of ovarian carcinoma cell lines, OVCAR-3, a low podocalyxin expressing cell line, was forced to overexpress the mouse podocalyxin cDNA. Forced overexpression of podocalyxin did not disrupt cellcell junctions in polarized monolayers. However, it did disrupt cohesive cell-cell aggregation in suspension. In addition, podocalyxin overexpression in depolarized monolayers disrupted celhsubtratum adhesion. Podocalyxin overexpression caused a marked decrease in cell adhesion to fibronectin, mesothelial cells and to anti-β1 integrin antibody-coated wells, suggesting that high levels of podocalyxin may interfere with integrin engagement. To investigate the consequences of podocalyxin overexpression on β1 integrin localization in polarized monolayers and 3D cell clusters, dual immunofluorescence staining was performed for podocalyxin/β1 integrin. Podocalxyin overexpressing monolayers targetted podocalyxin to the apical membrane, resulting in a subtle depletion of β1 integrin from this site. In non-polar 3D cultures, podocalyxin overexpressing cell clusters mislocalized podocalxyin to the cell-matrix interface causing β1 integrin depletion from this membrane domain. Therefore, as a cell-ECM disrupting anti-adhesin, mislocalized podocalyxin may promote the dissemination of primary tumour nodules into the peritoneal cavity by interfering with integrin engagement at the tumounECM interface.Medicine, Faculty ofCellular and Physiological Sciences, Department ofGraduat

Strategic Communications for Influence: Lessons From the Annie E. Casey Foundation and Its KIDS COUNT Initiative

· This article describes how the Annie E. Casey Foundation is using the KIDS COUNT Network in a new way: as a strategic communications tool in its focused efforts toward policy change, broad social change, and improved conditions for vulnerable children and families. An outcome map illustrates links between this strategy and the intended outcomes. · Case illustrations of KIDS COUNT grantee activities surrounding the release of the 2008 KIDS COUNT Data Book describe the efforts of grantees in six states where the quantity and quality of media coverage surrounding the national data book reflected the kind of coverage that Casey believes will help achieve its desired outcomes. · Strategic communications approaches such as relationships with journalists, use of locally relevant information, use of locally relevant media advocacy strategies, good preparation, and a solution orientation were present in states demonstrating desirable media coverage. · Prescribing specific communications tactics matters less than supporting the network’s general capacity to engage in year-round strategic communications approaches to create conditions (e.g., reputations, relationships) that will contribute to successful media advocacy related to a specific event such as the release of the national data book

The cell surface mucin podocalyxin regulates collective breast tumor budding

Background: Overexpression of the transmembrane sialomucin podocalyxin, which is known to play a role in lumen formation during polarized epithelial morphogenesis, is an independent indicator of poor prognosis in a number of epithelial cancers, including those that arise in the breast. Therefore, we set out to determine if podocalyxin plays a functional role in breast tumor progression. Methods: MCF-7 breast cancer cells, which express little endogenous podocalyxin, were stably transfected with wild type podocalyxin for forced overexpression. 4T1 mammary tumor cells, which express considerable endogenous podocalyxin, were retrovirally transduced with a short hairpin ribonucleic acid (shRNA) targeting podocalyxin for stable knockdown. In vitro, the effects of podocalyxin on collective cellular migration and invasion were assessed in two-dimensional monolayer and three-dimensional basement membrane/collagen gel culture, respectively. In vivo, local invasion was assessed after orthotopic transplantation in immunocompromised mice. Results: Forced overexpression of podocalyxin caused cohesive clusters of epithelial MCF-7 breast tumor cells to bud off from the primary tumor and collectively invade the stroma of the mouse mammary gland in vivo. This budding was not associated with any obvious changes in histoarchitecture, matrix deposition or proliferation in the primary tumour. In vitro, podocalyxin overexpression induced a collective migration of MCF-7 tumor cells in two-dimensional (2-D) monolayer culture that was dependent on the activity of the actin scaffolding protein ezrin, a cytoplasmic binding partner of podocalyxin. In three-dimensional (3-D) culture, podocalyxin overexpression induced a collective budding and invasion that was dependent on actomyosin contractility. Interestingly, the collectively invasive cell aggregates often contained expanded microlumens that were also observed in vivo. Conversely, when endogenous podocalyxin was removed from highly metastatic, but cohesive, 4T1 mammary tumor cells there was a decrease in collective invasion in three-dimensional culture. Conclusions: Podocalyxin is a tumor cell-intrinsic regulator of experimental collective tumor cell invasion and tumor budding.Medicine, Faculty ofOther UBCNon UBCCellular and Physiological Sciences, Department ofReviewedFacult

The cell surface mucin podocalyxin regulates collective breast tumor budding

Abstract Background Overexpression of the transmembrane sialomucin podocalyxin, which is known to play a role in lumen formation during polarized epithelial morphogenesis, is an independent indicator of poor prognosis in a number of epithelial cancers, including those that arise in the breast. Therefore, we set out to determine if podocalyxin plays a functional role in breast tumor progression. Methods MCF-7 breast cancer cells, which express little endogenous podocalyxin, were stably transfected with wild type podocalyxin for forced overexpression. 4T1 mammary tumor cells, which express considerable endogenous podocalyxin, were retrovirally transduced with a short hairpin ribonucleic acid (shRNA) targeting podocalyxin for stable knockdown. In vitro, the effects of podocalyxin on collective cellular migration and invasion were assessed in two-dimensional monolayer and three-dimensional basement membrane/collagen gel culture, respectively. In vivo, local invasion was assessed after orthotopic transplantation in immunocompromised mice. Results Forced overexpression of podocalyxin caused cohesive clusters of epithelial MCF-7 breast tumor cells to bud off from the primary tumor and collectively invade the stroma of the mouse mammary gland in vivo. This budding was not associated with any obvious changes in histoarchitecture, matrix deposition or proliferation in the primary tumour. In vitro, podocalyxin overexpression induced a collective migration of MCF-7 tumor cells in two-dimensional (2-D) monolayer culture that was dependent on the activity of the actin scaffolding protein ezrin, a cytoplasmic binding partner of podocalyxin. In three-dimensional (3-D) culture, podocalyxin overexpression induced a collective budding and invasion that was dependent on actomyosin contractility. Interestingly, the collectively invasive cell aggregates often contained expanded microlumens that were also observed in vivo. Conversely, when endogenous podocalyxin was removed from highly metastatic, but cohesive, 4T1 mammary tumor cells there was a decrease in collective invasion in three-dimensional culture. Conclusions Podocalyxin is a tumor cell-intrinsic regulator of experimental collective tumor cell invasion and tumor budding

Ekologické vinohradnictví a vinařství na Mikulovsku

This thesis is focused on organic viticulture and winemaking in Mikulov region. This work evaluates development of organic viticulture in Mikulov viniculture region and description of organic grape growing and organic viticulture. This evaluation is focused on the areas of individual municipalities, the number and size of farms and the production and trade of organic wine. We pay special attention to the description of interspecific vine varieties, the cultivation areas and selected varieties. Thanks to our investigations of organic wine growers and organic farming winemakers we can characterize the situation in the sector, describe their motivation to farming in organic mode and prosperity of organic wine market. 5.3% of vineyards in the Mikulov region are in the organic mode. We asked 8 of 23 organic farmers in the region. All of them are constant in organic mode mainly because of the quality of its production

Additional file 2: of The cell surface mucin podocalyxin regulates collective breast tumor budding

Is Movie S1 showing control MCF-7 cells present little collective invasion 3-D culture, and Movie S2 showing podo-overexpressing cells send out very dynamic processes and show considerable collective invasion in 3-D culture. (ZIP 2262 kb