To determine the relationship between yellow band disease (YBD)-associated pathogenic bacteria found in both Caribbean and Indo-Pacific reefs, and the virulence of these pathogens. YBD is one of the ...most significant coral diseases of the tropics. The consortium of four Vibrio species was isolated from YBD tissue on Indo-Pacific corals: Vibrio rotiferianus, Vibrio harveyi, Vibrio alginolyticus and Vibrio proteolyticus. This consortium affects Symbiodinium (zooxanthellae) in hospite causing symbiotic algal cell dysfunction and disorganization of algal thylakoid membrane-bound compartment from corals in both field and laboratory. Infected corals have decreased zooxanthella cell division compared with the healthy corals. Vibrios isolated from diseased Diploastrea heliopora, Fungia spp. and Herpolitha spp. of reef-building corals display pale yellow lesions, which are similar to those found on Caribbean Montastraea spp. with YBD. The Vibrio consortium found in YBD-infected corals in the Caribbean are close genetic relatives to those in the Indo-Pacific. The consortium directly attacks Symbiodinium spp. (zooxanthellae) within gastrodermal tissues, causing degenerated and deformed organelles, and depleted photosynthetic pigments in vitro and in situ. Infected Fungia spp. have decreased cell division compared with the healthy zooxanthellae: 4·9%vs 1·9%, (P greater-than-or-equal 0·0024), and in D. heliopora from 4·7% to 0·7% (P greater-than-or-equal 0·002). Pathogen virulence has major impacts on the survival of these important reef-building corals around the tropics.
Abstract Colorectal cancer (CRC) is the third most common carcinoma worldwide and despite advances in treatment, survival for patients with metastatic disease remains poor. With nearly 50% of ...patients developing metastases, in vivo investigation is essential to improve outcomes for these patients and numerous murine models of CRC have been developed to allow the study of chemoprevention and chemotherapy, in addition to improving our understanding of the pathogenesis of CRC. Selecting the most appropriate murine model for a specific application will maximize the conversion of potential therapies from the laboratory to clinical practice and requires an understanding of the various models available. This review will provide an overview of the murine models currently used in CRC research, discussing the limitations and merits of each and their most relevant application. It is aimed at the developing researcher, acting as a guide to prompt further reading in planning a specific study.
Nrf2 is a transcription factor that regulates cellular stress response and irinotecan-metabolising pathways. Its aberrant activity has been reported in a number of cancers, although relatively few ...studies have explored a role for Nrf2 in colorectal cancer (CRC). This study assessed the expression of Nrf2 in patient CRC tissues and explored the effect of Nrf2 modulation alone, or in combination with irinotecan, in human (HCT116) and murine (CT26) cell lines
and in an orthotopic syngeneic mouse model utilising bioluminescent imaging. Using a tissue microarray, Nrf2 was found to be overexpressed (p<0.01) in primary CRC and metastatic tissue relative to normal colon, with a positive correlation between Nrf2 expression in matched primary and metastatic samples.
experiments in CRC cell lines revealed that Nrf2 siRNA and brusatol, which is known to inhibit Nrf2, decreased viability and sensitised cells to irinotecan toxicity. Furthermore, brusatol effectively abrogated CRC tumour growth in subcutaneously and orthotopically-allografted mice, resulting in an average 8-fold reduction in luminescence at the study end-point (p=0.02). Our results highlight Nrf2 as a promising drug target in the treatment of CRC.
Improving outcomes in colorectal cancer requires more accurate in vivo modelling of the disease in humans, allowing more reliable pre-clinical assessment of potential therapies. Novel imaging ...techniques are necessary to improve the longitudinal assessment of disease burden in these models, reducing the number of animals required for translational studies. This report describes the development of an immune-competent syngeneic orthotopic murine model of colorectal cancer, utilising caecal implantation of CT26 cells stably transfected with the luciferase gene into immune-competent BALB/c mice, allowing serial bioluminescent imaging of cancer progression. Luminescence in the stably transfected CT26 cell line, after pre-conditioning in the flank of a BALB/c mouse, accurately reflected cell viability and resulted in primary caecal tumours in five of eight (63%) mice in the initial pilot study following caecal injection. Luminescent signal continued to increase throughout the study period with one mouse (20%) developing a liver metastasis. Histopathological assessment confirmed tumours to be consistent with a poorly differentiated adenocarcinoma. We have now performed this technique in 68 immune-competent BALB/c mice. There have been no complications from the procedure or peri-operative deaths, with primary tumours developing in 44 (65%) mice and liver metastases in nine (20%) of these. This technique provides an accurate model of colorectal cancer with tumours developing in the correct microenvironment and metastasising to the liver with a similar frequency to that seen in patients presenting with colorectal cancer, with serial bioluminescent reducing the murine numbers required in studies by removing the need for cull for assessment of disease burden.
Epithelial ovarian cancer (EOC) metastasizes transcoelomically to the peritoneum and omentum, and despite surgery and chemotherapy, recurrent disease is likely. Metastasis requires the induction of ...proangiogenic changes in the omental microenvironment and EOC-induced omental angiogenesis is currently a key therapeutic target. In particular, antiangiogenic therapies targeting the vascular endothelial growth factor A (VEGFA) pathway are commonly used, although, with limited effects. Here, using human omental microvascular endothelial cells (HOMECs) and ovarian cancer cell lines as an in vitro model, we show that factors secreted from EOC cells increased proliferation, migration, and tube-like structure formation in HOMECs. However, EOC-induced angiogenic tube-like formation and migration were unaffected by inhibition of tyrosine kinase activity of VEGF receptors 1 and 2 (Semaxanib; SU5416) or neutralization of VEGFA (neutralizing anti-VEGFA antibody), although VEGFA165 -induced HOMEC migration and tube-like structure formation were abolished. Proteomic investigation of the EOC secretome identified several alternative angiogenesis-related proteins. We screened these for their ability to induce an angiogenic phenotype in HOMECs, i.e., proliferation, migration, and tube-like structure formation. Hepatocyte growth factor (HGF) and insulin-like growth factor binding protein 7 (IGFBP-7) increased all three parameters, and cathepsin L (CL) increased migration and tubule formation. Further investigation confirmed expression of the HGF receptor c-Met in HOMECs. HGF- and EOC-induced proliferation and angiogenic tube structure formation were blocked by the c-Met inhibitor PF04217903. Our results highlight key alternative angiogenic mediators for metastatic EOC, namely, HGF, CL, and IGFBP-7, suggesting that effective antiangiogenic therapeutic strategies for this disease require inhibition of multiple angiogenic pathways.
Abstract Epithelial ovarian cancer (EOC) metastasis to the omentum requires implantation and angiogenesis. We propose that prometastatic changes in the omental endothelium (for angiogenesis) and ...mesothelium (for implantation) are critical. We investigated the expression of angiogenic proteases cathepsin D (CD), cathepsin L (CL), and matrix metalloproteinase 2 (MMP2) and MMP9 and vascular endothelial growth factor A (VEGFA) in the mesothelium and endothelium of omentum from patients with EOC with omental metastases and control patients with benign ovarian tumors. Endothelial expression of CL, VEGFA, and MMP9 and mesothelial expression of VEGFA, MMP9, and CD were significantly increased in patients with metastasized EOC. High expression of MMP9 and VEGFA in endothelium and mesothelium and CD in mesothelium was positively associated with poor disease-specific survival (DSS). High MMP9 expression in either endothelium or mesothelium and presence of ascites prospectively showed the greatest risk of shorter DSS hazard ratio (HR)= 6.16, 95% confidence interval (CI) = 1.76-21.6, P = .0045; HR = 11.42, 95% CI = 2.59-50.35, P = .0013; and HR = 6.35, 95% CI = 2.01-20.1, P = .002, respectively. High endothelial MMP9 expression and ascites were independent predictors of reduced DSS and overall survival, together resulting in worst patient prognosis. Our data show that omental metastasis of EOC is associated with increased proangiogenic protein expression in the omental endothelium and mesothelium.
Please cite this paper as: Winiarski, Acheson, Gutowski, McHarg and Whatmore (2011). An Improved and Reliable Method for Isolation of Microvascular Endothelial Cells from Human Omentum. ...Microcirculation18(8), 635–645.
Objectives: Despite an increasing research demand for human microvascular endothelial cells, isolation of primary endothelial cells from human tissue remains difficult. The omentum, a highly vascular visceral adipose tissue, could provide an excellent source of these cells.
Methods: A reliable method to isolate HOMECs has been developed. It consists of initial enzymatic digestion (to deplete cell contaminants), followed by further digestion, selective filtration, and immunoselection using Dynabeads coated with CD31 antibody. Cultures were characterized for expression of endothelial cell markers and their ability to undergo VEGF‐dependent in vitro tube structure formation.
Results: Omental‐derived cultures of microvascular endothelial cells were achieved with <5% contamination of other cell types. The endothelial origin of cells was confirmed by the constitutive expression of a range of vascular endothelial markers (CD31, CD105, vWF) and internalization of DiI‐AcLDL. Furthermore, cultures were negative for lymphatic endothelial markers, underwent in vitro angiogenesis, and exhibited typical endothelial morphology.
Conclusions: This isolation method produces homogeneous HOMEC cultures that can be maintained in vitro for at least six passages without loss of cellular features characterizing endothelial cells.
Spatially explicit model of wintering common loons Winiarski, Kristopher J.; Miller, David L.; Paton, Peter W. C. ...
Marine ecology. Progress series (Halstenbek),
10/2013, Letnik:
492
Journal Article
Recenzirano
Odprti dostop
Common loons Gavia immer are a conservation concern in New England due to a variety of anthropogenic factors, yet little is known about biotic and abiotic environmental factors determining their ...wintering distribution and abundance in nearshore and offshore waters. The primary objective of this study was to develop a spatially explicit abundance model of wintering common loons in the maritime waters of southern New England (USA) that could inform decisions about offshore development. Aerial line-transect surveys were conducted throughout a 3800 km² study area off the coast of Rhode Island during the winters of 2010–2011 and 2011–2012. A density surface model (DSM) approach was used to account for imperfect detection and incorporate spatially explicit environmental covariates. Common loon densities were greatest in waters <35 m deep, with high chl a surface concentrations (>2 mg m–3). The DSM predicted 5047 (95% CI = 3993–6379) common loons in the study area during winter, which suggests this region provides key habitat for this species in eastern North America. This study highlights important areas for common loons in the region, suggests key biotic (primary productivity as measured by long-term chl a surface concentrations) and abiotic covariates (water depth) driving the spatial distribution and abundance of common loons in southern New England, and identifies sites that should be considered for protection from offshore development, including offshore wind facilities.
Seabirds are of conservation concern, and as new potential risks to seabirds are arising, the need to provide unbiased estimates of species’ distributions is growing. We applied community occupancy ...models to detection/non-detection data collected from repeated aerial strip-transect surveys conducted in 2 large study plots off southern New England, USA; one off the coast of Rhode Island and the other in Nantucket Sound. A total of 17 seabird species were observed at least once in each study plot. We found that detection varied by survey date and effort for most species and the average detection probability across species was less than 0.4. We estimated the influence of water depth, sea surface temperature, and sea surface chl a concentration on species-specific occupancy. Diving species showed large differences between the 2 study plots in their predicted winter distributions, which were largely explained by water depth acting as a stronger predictor of occupancy in Rhode Island than in Nantucket Sound. Conversely, similarities between the 2 study plots in predicted winter distributions of surface-feeding species were explained by sea surface temperature or chlorophyll a concentration acting as predictors of these species’ occupancy in both study plots. We predicted the number of species at each site using the observed data in order to detect ‘hot-spots’ of seabird diversity and use in the 2 study plots. These results provide new information on detection of species, areas of use, and relationships with environmental variables that will be valuable for biologists and planners interested in seabird conservation in the region.
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