Mesenchymal Stem Cells Promote Mammosphere Formation and Decrease E-Cadherin in Normal and Malignant Breast Cells

Normal and malignant breast tissue contains a rare population of multi-potent cells with the capacity to self-renew, referred to as stem cells, or tumor initiating cells (TIC). These cells can be enriched by growth as "mammospheres" in three-dimensional cultures.We tested the hypothesis that human bone-marrow derived mesenchymal stem cells (MSC), which are known to support tumor growth and metastasis, increase mammosphere formation.We found that MSC increased human mammary epithelial cell (HMEC) mammosphere formation in a dose-dependent manner. A similar increase in sphere formation was seen in human inflammatory (SUM149) and non-inflammatory breast cancer cell lines (MCF-7) but not in primary inflammatory breast cancer cells (MDA-IBC-3). We determined that increased mammosphere formation can be mediated by secreted factors as MSC conditioned media from MSC spheroids significantly increased HMEC, MCF-7 and SUM149 mammosphere formation by 6.4 to 21-fold. Mammospheres grown in MSC conditioned media had lower levels of the cell adhesion protein, E-cadherin, and increased expression of N-cadherin in SUM149 and HMEC cells, characteristic of a pro-invasive mesenchymal phenotype. Co-injection with MSC in vivo resulted in a reduced latency time to develop detectable MCF-7 and MDA-IBC-3 tumors and increased the growth of MDA-IBC-3 tumors. Furthermore, E-cadherin expression was decreased in MDA-IBC-3 xenografts with co-injection of MSC.MSC increase the efficiency of primary mammosphere formation in normal and malignant breast cells and decrease E-cadherin expression, a biologic event associated with breast cancer progression and resistance to therapy

Mechanisms of leukocyte lipid body formation and function in inflammation

An area of increasingly interest for the understanding of cell signaling are the spatio-temporal aspects of the different enzymes involved in lipid mediator generation (eicosanoid-forming enzymes, phospholipases and their regulatory kinases and phosphatases) and pools of lipid precursors. The compartmentalization of signaling components within discrete and dynamic sites in the cell is critical for specificity and efficiency of enzymatic reactions of phosphorilation, enzyme activation and function. We hypothesized that lipid bodies - inducible non-membrane bound cytoplasmic lipid domains - function as specialized intracellular sites of compartmentalization of signaling with major roles in lipid mediator formation within leukocytes engaged in inflammatory process. Over the past years substantial progresses have been made demonstrating that all enzymes involved in eicosanoid synthesis localize at lipid bodies and lipid bodies are distinct sites for eicosanoid generation. Here we will review our current knowledge on the mechanisms of formation and functions of lipid bodies pertinent to inflammation

A prospective pilot study comparing combined intravitreal ranibizumab and half-fluence photodynamic therapy with ranibizumab monotherapy in the treatment of neovascular age-related macular degeneration

Patrick D Williams,1 David Callanan,1 Wayne Solley,1 Robert L Avery,2 Dante J Pieramici,2 Tom Aaberg31Texas Retina Associates, Dallas, TX, 2California Retina Consultants and Research Foundation, Santa Barbara, CA, 3Retinal Associates, Grand Rapids, MI, USAPurpose: This prospective multi-center pilot study compares the use of half-fluence photodynamic therapy combined with ranibizumab with ranibizumab monotherapy for the treatment of neovascular age-related macular degeneration.Methods: All patients presenting with untreated subfoveal neovascular age-related macular degeneration were considered for inclusion. Patients were randomized to receive either ranibizumab with half-fluence photodynamic therapy or ranibizumab alone. Patients in the ranibizumab alone group were given three consecutive monthly ranibizumab injections and were followed monthly. They were treated with ranibizumab as needed, based on clinical discretion, using vision and optical coherence tomography. Patients in the combined group were given one same-day combined ranibizumab and half-fluence (25 j/cm2) photodynamic therapy treatment and were treated monthly as needed. Outcomes included changes in standardized visual acuity, optical coherence tomography foveal thickness, and percentage of as-needed injections to maintenance examinations.Results: Fifty-six out of 60 enrolled patients completed the twelve month primary outcome visit; this consisted of 27 patients receiving ranibizumab alone and 29 receiving combined treatment. The average age was 79.1 for the ranibizumab alone group and 79.3 for the combined group. The mean visual acuity in the ranibizumab alone group improved from 52.9 Early Treatment of Diabetic Retinopathy letters initially to 62.8 letters at twelve months. The mean visual acuity in the combined group improved from 49.2 letters to 51.8 letters at twelve months. The differences in visual acuity improvements were not statistically significant based on a two-tailed t-test (P = 0.2). Due to the presence of outliers in each group, a Mann–Whitney U test was performed to confirm the results (U = 325; P = 0.28). The mean optical coherence tomography foveal thickness improved 92.5 microns and 106.7 microns in the ranibizumab alone and the combined group, respectively. The difference was not significant based on a two-tailed t-test (P = 0.6). The ranibizumab alone group received an average of 6.8 injections, while the combined group received an average of three injections. This difference was not significant based on a chi-square test (P = 0.11).Conclusion: The groups appeared similar based on statistical analysis, but larger studies are needed to determine possible small differences between combination therapy and monotherapy.Keywords: ranibizumab, macular degeneration, photodynamic therapy, verteporfin, choroidal neovascularization, anti-veg

Identifying novel wastewater treatment options through optimal technology integration

This paper presents the outcomes of the first of four phases of a research project which aims to investigate the optimal integration of novel, yet largely already demonstrated technologies, in water recycling process train options. The research project has a particular focus on the minimisation of environmental and economic costs in the implementation and operation of the overall recycling process. In the Phase 1 desktop study, novel carbon and nitrogen removal technologies were selected through multi-criteria analysis and incorporated into two concept stage integrated treatment train options. The first treatment train option includes a main-stream anaerobic membrane bioreactor, followed by soluble methane stripping. Nitrogen removal is then achieved with a nitritation/Anammox moving bed biofilm reactor. The second alternative treatment train utilises a high rate/solids contact activated sludge system, with the separated excess biomass treated in a two-stage high-rate anaerobic sludge digester. The digested biosolids stream is treated separately to recover nutrients (as struvite) and to remove nitrogen with a nitritation/Anammox process. Further mainstream nitrogen removal is achieved with a nitrification/denitrification process in a sequencing batch reactor configuration. The two alternative treatment trains were compared to a typical existing treatment train for economics and environmental footprint under Australian conditions at two scales; 10 and 100 ML/d average flow. Engineering analysis included high-level concept design and sizing, estimates of performance, assessment of environmental footprint and whole-of-life cost estimates (including capital and operating expenses). The results of the study indicated that the new treatment trains have the potential to significantly decrease the economic costs of wastewater treatment by between 10 and 46% (based on Net Present Value estimates), and have a lower environmental impact. In Phases 2 and 3, lab-scale and pilot studies are currently underway to further evaluate the performance and confirm the design/operating parameters of the core processes

Attenuation of groundwater pollution by bank filtration

Bank filtration, either natural or induced through the river bed by pumping from a system of connected lateral or vertical wells, provides a means of obtaining public water supplies. The success of such schemes is dependent on the microbial activity and chemical transformations that are commonly enhanced in the colmation layer within the river bed compared to those that take place in surface or ground waters. The actual biogeochemical interactions that sustain the quality of the pumped bank filtrate depend on numerous factors including aquifer mineralogy, shape of the aquifer, oxygen and nitrate concentrations in the surface water, types of organic matter in the surface and ground water environments, and land use in the local catchment area. This paper provides an introduction to a series of nine papers contained in this Special Issue that highlight these factors and finishes with a list of recommendations for co-ordinated research into attenuation of groundwater pollution by bank filtration