A Novel Cartesian Plot Analysis for Fixed Monolayers That Relates Cell Phenotype to Transfer of Contents between Fibroblasts and Cancer Cells by Cell-Projection Pumping

We recently described cell-projection pumping as a mechanism transferring cytoplasm between cells. The uptake of fibroblast cytoplasm by co-cultured SAOS-2 osteosarcoma cells changes SAOS-2 morphology and increases cell migration and proliferation, as seen by single-cell tracking and in FACS separated SAOS-2 from co-cultures. Morphological changes in SAOS-2 seen by single cell tracking are consistent with previous observations in fixed monolayers of SAOS-2 co-cultures. Notably, earlier studies with fixed co-cultures were limited by the absence of a quantitative method for identifying sub-populations of co-cultured cells, or for quantitating transfer relative to control populations of SAOS-2 or fibroblasts cultured alone. We now overcome that limitation by a novel Cartesian plot analysis that identifies individual co-cultured cells as belonging to one of five distinct cell populations, and also gives numerical measure of similarity to control cell populations. We verified the utility of the method by first confirming the previously established relationship between SAOS-2 morphology and uptake of fibroblast contents, and also demonstrated similar effects in other cancer cell lines including from melanomas, and cancers of the ovary and colon. The method was extended to examine global DNA methylation, and while there was no clear effect on SAOS-2 DNA methylation, co-cultured fibroblasts had greatly reduced DNA methylation, similar to cancer associated fibroblasts.     

Remote and in situ photogrammetric history of the Kidd Copper tailings site,Sudbury, Ontario

There is much concern regarding the genesis of acidic leachate by mine tailings and its potential impact on local water systems. Genesis of the leachate and its subsequent escape into the surrounding watershed is controlled by the primary distribution of facies within the tailings pond. Leachate generation and percolation is enhanced in high-sulphide coarse-grained sands, while percolation is restricted in fine-grained clays formed from bentonitic slimes. The Kidd Copper deposit, located on the South Range of the Sudbury Basin, was a typical offset dike nickel deposit, where mining took place between 1970 and 1990. Using a sequence of temporally separate aerial photographs dating from 1945 to 2002, it is possible to document the initial development of the mine, its closure and the subsequent degradation of the mine tailings waste that was produced during mining activity. Since the mine's closure, the resulting tailings deposit has progressively oxidized, generating acid-rich leachate. Leachate can escape from the tailings via surface run-off and subsurface flow. The spatial distribution of possible flow pathways was mapped using a combination of sub-centimetre-resolution real-time kinematic (RTK) GPS and individual, high-resolution, oriented photographs of subsurface pits. Lithologic mapping information of the Kidd Copper tailings deposit can provide an insight into the distribution of local aquifers and aquitards. Subsurface lithologic mapping was achieved using a combination of sub-centimetre-resolution RTK GPS and individual, high-resolution, oriented photographs of subsurface pits. The photograph of each pit provides lithologic information of a specific point in the tailings deposit, while the high-resolution GPS locates the individual photographs in three-dimensional (3D) geographic space. Combining these data allows us to identify continuous lithologic surfaces between pits, leading to the construction of a 3D model of the deposition of the tailings pond. Grain size and textural details extracted from the images of each lithology permit discrimination between aquifers and aquitards. The final geometric model could be used as input for estimating the hydrologic and chemical evolution of the tailings.     

Generation of Free Radicals in RDX and HMX Compositions

Efforts to generate detectable concentrations of free radicals in explosives, binders, and their mixtures are described. Radicals were readily produced in polycaprolactone and polyethylene glycol binders at liquid nitrogen temperature using stresses as low as 0.4 kbar. These radicals were all of the peroxy type, and presumably formed by reaction of mechano-radicals with oxygen present in the polymer. No mechano-radicals were observed from HMX or RDX using samples cooled to liquid nitrogen temperature and applied stresses up to 4 kbar. In neither impacted samples that failed to explode nor the residues remaining after a partial explosion were radicals detected by ESR. Low temperature γ-irradiation of these materials was also carried out. Free radical signals originating in both the polymer and the explosive could be identified. The reactivity of NO₂ radicals from γ-irradiated HMX is enhanced in the presence of binder. In γ-irradiated HMX/polycaprolactone mixtures, the NO₂ radical anneals rapidly at 150 K, versus 240 K for HMX alone. Evidence is presented to suggest that the relatively stable NO₂ radical (such as produced by γ-irradiation) in HMX does not play a dominant role in mechanical initiation.     

Reinventing Reality: Reconstructing Architecture

This article is based on a paper originally given as keynote address at the Ontario Association of Architects Convention, Toronto, March, 1988. Three issues are explored in this article: the historical background to the present situation in architecture and architectural education, with special reference to the work of Christian Norberg-Schulz, Robert Venturi, Colin Rowe, Bruno Zevi, Peter Eisenman and Michael Graves; the value of focussing again on the agenda of Norberg-Schulz's Intentions in Architecture (1962) as being a more reliable guide than the scattered opinions offered by other architects; a strategy for redesigning architectural education to reflect Norberg-Schulz's fundamental concerns about the role of architecture and architects, offering an integrated curriculum based upon a vertical rather than a horizontal structure of learning.     

Quantifying the extent of space shortages: English dwellings

Lack of internal space is a problem in UK homes, and is often linked to a lack of space standards for housing in the UK. Although previous studies have examined new-build housing, this paper uses a new method to study 16 000 dwellings in the English Housing Survey (EHS) 2010 to examine existing housing and compare them with a modern space standard in an attempt to quantify the extent and magnitude of the problem. Dwellings in the survey were compared against a slightly modified version of the London Housing Design Guide 2010 internal space standard. It was found that between 21% and 55% of dwellings failed to meet the standard; and that flats and small terraced houses were most commonly below the standard. Dwellings were also frequently found to be under-occupied in comparison with the number of bedrooms. This research hypothesized that this was a result of the lack of space. To illustrate the use of this new analysis method, a recent change to the Housing Benefit system (colloquially known as the ‘Bedroom Tax’) has been considered. Households receiving Housing Benefit were more likely to be undersized, suggesting that the policy to withdraw housing benefits from these households may be misguided.     

Treatment of an Iron-rich ARD Using Waste Carbonate Rock: Bench-Scale Reactor Test Results

The treatment of acid rock drainage (ARD) places extraordinary financial burdens on governments and companies worldwide, and an improved efficiency in treatment by as little as 1% can save many millions of dollars in rehabilitation. We investigated a system for treating Fe-rich ARD using a three-stage reactor design. In the first reaction cell, Fe-rich ARD was partially neutralised using rapid periodic carbonate resuspension with a rotating axial mixer. This was followed by an air-sparged oxidation chamber and then a second reaction cell, with more carbonate periodically resuspended until a pH of 6.3 was reached, which was followed by a settlement chamber. This reactor design has a high capacity for neutralisation, with an efficiency of ≈70% of acidity neutralised by the acid neutralising capacity (g of CaCO₃ equivalent) added to the reactor. Axial mixers were tested because of their low-energy requirements and their high reliability. The intermediate chamber effectively removes Fe by oxidising Fe(II) to Fe(III). Given the amount of acidity neutralised, the sludge volume produced was low compared to other technologies, providing further potential savings in sludge handling. Waste carbonate rock proved to be an effective neutralising agent, even though it was about 60% dolomite and 40% magnesite, with minor calcite, and despite the fact that magnesite has substantially slower dissolution kinetics compared to the more dominant dolomite. The mixed waste carbonates were capable of raising the pH sufficiently to reduce the heavy metal loadings in Fe-rich ARD by more than two orders of magnitude. The final settlement stage of the process was shown to be essential for metal precipitation, for the carry-over of fine carbonates, and CO₂ loss. This was associated with a rise in pH, from 6.3 to 7.5. In addition, residual slow-reacting magnesite from the mixed carbonate remains in the sludge from the first reactor and provides acid buffering capacity within the sludge, which is commonly lacking in the ARD neutralisation sludge of other systems.