Paracrine signaling through receptor activator of NF-kB (RANK) pathway mediates the expansion of mammary epithelia that occurs during pregnancy, and activation of RANK pathway promotes mammary tumorigenesis in mice. In this study we extend these previous data to human cells and show that the RANK pathway promotes the development of mammary stem cells and breast cancer. Overexpression of RANK (FL-RANK) in a panel of tumoral and normal human mammary cells induces the expression of breast cancer stem and basal/stem cell markers. High levels of RANK in untransformed MCF10A cells induce changes associated with both stemness and transformation, including mammary gland reconstitution, epithelial-mesenchymal transition (EMT), increased migration, and anchorage-independent growth. In addition, spheroids of RANK overexpressing MCF10A cells display disrupted acinar formation, impair growth arrest and polarization, and luminal filling. RANK overexpression in tumor cells with nonfunctional BRCA1 enhances invasiveness in acinar cultures and increases tumorigenesis and metastasis in immunodeficient mice. High levels of RANK were found in human primary breast adenocarcinomas that lack expression of the hormone receptors, estrogen and progesterone, and in tumors with high pathologic grade and proliferation index; high RANK/RANKL expression was significantly associated with metastatic tumors. Together, our findings show that RANK promotes tumor initiation, progression, and metastasis in human mammary epithelial cells by increasing the population of CD44 þ CD24À cells, inducing stemness and EMT. These results suggest that RANK expression in primary breast cancer associates with poor prognosis. Cancer Res; 72(11); 2879-88. Ó2012 AACR.

We investigated convection caused by surface cooling and mixing attributable to wind shear stress and their roles as agents for the transport of phytoplankton cells in the water column by carrying out two daily surveys during the stratified period of the Sau reservoir. Green algae, diatoms, and cryptophyceae were the dominant phytoplankton communities during the surveys carried out in the middle (July) and end (September) of the stratified period. We show that a system with a linear stratification and that is subject to weak surface forcing, with weak winds ,4 m s 21 and low energy dissipation rate values of the order of 10 28 m 2 s 23 or lower, enables the formation of thin phytoplankton layers. These layers quickly disappear when water parcels mix because there is a medium external forcing (convection) induced by the night surface cooling, which is characterized by energy dissipation rates on the order of ,5 3 10 28 m 2 s 23 . During both surveys the wind generated internal waves during the entire diurnal cycle. During the day, and because of the weak winds, phytoplankton layers rise in the water column up to a depth determined by both solar heating and internal waves. In contrast, during the night phytoplankton mixes down to a depth determined by both convection and internal waves. These internal waves, together with the wind-driven current generated at the surface, seem to be the agents responsible for the horizontal transport of phytoplankton across the reservoir.

Most malignant MP breast tumours are variants of MBC. MP-MBC with predominant mesenchymal components behaves similar to ductal carcinomas and although data on their response to systemic therapy is limited, there is no evidence that they should be managed differently from other forms of triple negative breast cancer.

No abstract

Fixed weights moorings, once removed, can create longitudinal gaps in seagrass meadows of different sizes, running perpendicular to the coast. We quantified the interactions between these longitudinal gaps and the hydrodynamic environment of the nearshore environment to determine their potential impact on seagrass meadow ecology. Within the meadow at leaf length distances from the edge, wave attenuation by the lateral vegetation next to the gap was approximately the same as attenuation by fully vegetated areas, and the wave attenuating capacity of the lateral, near-gap vegetation was independent of gap width. Gaps with widths less than twice the leaf length exhibited 8% wave attenuation and 11% turbulent kinetic energy attenuation, confirming that vegetation shelters at least small gaps. Despite similar capacity for wave attenuation, the longitudinal gaps influenced the architectural characteristics of the adjacent (lateral) meadow; lateral shoot density, percent cover and leaf length adjacent to the largest gap were 12, 16, and 20% lower than the fully vegetated site, respectively. Significant differences in the temporal variation of the mean lateral, near-gap seagrass percent cover and the leaf length indicated a strong dependence of the state of the canopy on temporal hydrodynamic conditions, which in turn were impacted by the presence of the gap. Our results quantify the interactions between gaps and lateral meadow vegetation, highlight the structural impact of traditional moorings and support improved management and conservation of seagrass meadows.

The systematic creation of gaps within canopies results in fragmentation and the architecture of fragmented canopies differs substantially from non-fragmented canopies. Canopy fragmentation leads to spatial heterogeneity in hydrodynamics and therefore heterogeneity in the sheltering of canopy communities. Identifying the level of instability due to canopy fragmentation is important for canopies in coastal areas impacted by human activities and indeed, climate change. The gap orientation relative to the wave direction is expected to play an important role in determining wave attenuation and sheltering. Initially we investigated the effect of a single transversal gap within a canopy (i.e. a gap oriented perpendicular to the wave direction) on hydrodynamics, which was compared to fully vegetated canopies (i.e. no gaps) and also to bare sediment. The wave velocity increased with gap width for the two canopy densities studied (2.5% and 10% solid plant fraction) reaching wave velocities found over bare sediments. The turbulent kinetic energy (TKE) within the gap also increased, but was more attenuated by the adjacent vegetation than the wave velocity. As expected, denser canopies produced a greater attenuation of both the wave velocity and the turbulent kinetic energy within an adjacent gap, compared to sparse canopies. Using non-dimensional analysis and our experimental data, a parameterization for predicting TKE in a canopy gap was formulated, as a function of easily measured variables. Based on the experimental results, a fragmented canopy model was then developed to determine the overall mixing level in such canopies. The model revealed that canopies with large gaps present more mixing than canopies with small gaps despite having the same total gap area in the canopy. Furthermore, for the same total gap area, dense fragmented canopies provide more shelter than sparse fragmented canopies.

A laboratory flume experiment was carried out in which the hydrodynamic and sedimentary behaviour of a turbidity current was measured as it passed through an array of vertical rigid cylinders. The cylinders were intended primarily to simulate aquatic vegetation canopies, but could equally be taken to represent other arrays of obstacles, for example forests or offshore wind turbines. The turbidity currents were generated by mixing naturally sourced, poly‐disperse sediment into a reservoir of water at concentrations from 1·0 to 10·0 g l−1, which was then released into the experimental section of the flume by removing a lock gate. For each initial sediment concentration, runs with obstacle arrays with solid plant fractions of 1·0% and 2·5%, and control cases with no obstacles, were carried out. The progress of the current along the flume was characterized by the array drag term, CDaxc (where CD is the array drag coefficient, at the frontal area of cylinders per unit volume, and xc is the position of the leading edge of the current along the flume). The downward depositional flux of sediment out of the current as it proceeded was measured at 13 traps along the flume. Analysis of these deposits divided them into fine (2·2 to 6·2 μm) and coarse (6·2 to 104 μm) fractions. At the beginning of their development, the gravity currents proceeded in an inertia‐dominated regime until CDaxc = 5. For CDaxc > 5, the current transitioned into a drag‐dominated regime. For both fine and coarse sediment fractions, the rate of sediment deposition tended to decrease gradually with distance from the source in the inertial regime, remained approximately constant at the early drag‐dominated regime, and then rose and peaked at the end of the drag‐dominated stage. This implies that, when passing through arrays of obstacles, the turbidity currents were able to retain sufficient sediment in suspension to maintain their flow until they became significantly influenced by the drag exerted by the obstacles.

IntroductionEndocrine therapies targeting cell proliferation and survival mediated by estrogen receptor α (ERα) are among the most effective systemic treatments for ERα-positive breast cancer. However, most tumors initially responsive to these therapies acquire resistance through mechanisms that involve ERα transcriptional regulatory plasticity. Herein we identify VAV3 as a critical component in this process.MethodsA cell-based chemical compound screen was carried out to identify therapeutic strategies against resistance to endocrine therapy. Binding to ERα was evaluated by molecular docking analyses, an agonist fluoligand assay and short hairpin (sh)RNA–mediated protein depletion. Microarray analyses were performed to identify altered gene expression. Western blot analysis of signaling and proliferation markers, and shRNA-mediated protein depletion in viability and clonogenic assays, were performed to delineate the role of VAV3. Genetic variation in VAV3 was assessed for association with the response to tamoxifen. Immunohistochemical analyses of VAV3 were carried out to determine its association with therapeutic response and different tumor markers. An analysis of gene expression association with drug sensitivity was carried out to identify a potential therapeutic approach based on differential VAV3 expression.ResultsThe compound YC-1 was found to comparatively reduce the viability of cell models of acquired resistance. This effect was probably not due to activation of its canonical target (soluble guanylyl cyclase), but instead was likely a result of binding to ERα. VAV3 was selectively reduced upon exposure to YC-1 or ERα depletion, and, accordingly, VAV3 depletion comparatively reduced the viability of cell models of acquired resistance. In the clinical scenario, germline variation in VAV3 was associated with the response to tamoxifen in Japanese breast cancer patients (rs10494071 combined P value = 8.4 × 10−4). The allele association combined with gene expression analyses indicated that low VAV3 expression predicts better clinical outcome. Conversely, high nuclear VAV3 expression in tumor cells was associated with poorer endocrine therapy response. Based on VAV3 expression levels and the response to erlotinib in cancer cell lines, targeting EGFR signaling may be a promising therapeutic strategy.ConclusionsThis study proposes VAV3 as a biomarker and a rationale for its use as a signaling target to prevent and/or overcome resistance to endocrine therapy in breast cancer.