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.     

Visual Notes and the Acquisition of Architectural Knowledge

This article discusses the value and purpose of drawing in its relationship to the design process and understanding architecture. The authors base much of their discussion on their own experiences in teaching both drawing and architectural design to undergraduates. These experiences however are informed and supported both by architectural history and theory and theories of perception and art. The authors discuss the importance of visual notes to architects of the past, note both the decline of the usage of visual notes in the twentieth century as well as their continued use by important architects of our time. This article stresses the analytical and critical value of such drawing as opposed to its pictorial purposes.     

Influence of a wide range of calcium intakes on tissue distribution of macroelements and microelements in dairy calves

Sixteen intact male Holstein calves averaging 86 kg and 63 d of age were assigned randomly to four treatment groups. The four treatment diets contained .17, .67, 1.31, and 2.35% Ca on an as-fed basis. The resulting Ca:P ratios with P held constant at about .34% were .47:1, 1.92:1, 3.83:1, and 7.20:1. Calves were fed diets at 3% of their body weights for 4 wk. Magnesium in the bone ash and serum was lowered by the 2.35% Ca treatment. Serum inorganic P was also reduced by the highest Ca diet during the last 2 wk of the experiment. Liver had the highest concentration of Zn in calves fed .67% Ca, and the muscle from calves fed 1.31% Ca diet had the lowest amount of Zn. Copper was reduced in pancreas for 1.31% Ca diet, but Ca was highest in the muscle and heart at the .67% Ca treatment. Weight gains and feed efficiencies were not affected by Ca. Fecal pH was different among treatments and increased as Ca intake increased. Young growing dairy calves can adapt to a wide range of Ca intakes and Ca:P ratios and maintain a moderate growth rate for 4 wk. It appears that excessive dietary Ca may affect concentrations of Zn, Fe, Cu, and Mn in some body tissues, but the magnitude of the effect is relatively small.     

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.     

Surface-based Transport Model for Mixed-Size Sediment

We present a transport model for mixed sand/gravel sediments. Fractional transport rates are referenced to the size distribution of the bed surface, rather than subsurface, making the model completely explicit and capable of predicting transient conditions. The model is developed using a new data set of 48 coupled observations of flow, transport, and bed surface grain size using five different sediments. The model incorporates a hiding function that resolves discrepancies observed among earlier hiding functions. The model uses the full size distribution of the bed surface, including sand, and incorporates a nonlinear effect of sand content on gravel transport rate not included in previous models. The model shares some common elements with two previous surface-based transport models, but differs in using the full surface size distribution and in that it is directly developed from a relatively comprehensive data set with unambiguous measurement of surface grain size over a range of flow, transport rate, and sediments.     

Measurement of the density and surface tension of Ni-based superalloys in the liquid and mushy states

The densities of three Ni-based superalloys have been measured in both liquid and mushy states by both a modified sessile drop method (MSDM) and a modified pycnometric method (MPM) for alloys CMSX-4 and CM186LC, and for CMSX-10 alloy by MSDM only. The surface tensions of liquid CMSX-4, CM186LC, and CMSX-10 superalloys were measured using the sessile drop method. All measurements were carried out in a highly purified argon atmosphere with the oxygen partial pressure of less than 10⁻¹⁹ MPa in the gas outlet. The densities of all superalloys in both liquid and mushy states were found to decrease with increasing temperature. The volume thermal expansion of each superalloy in the mushy state was found to be higher than that in the liquid state. The densities determined by different methods have been critically assessed and recommended values in both liquid and mushy states are given as a linear function of temperature for the three Ni-based superalloys. The surface tension of liquid CMSX-4 superalloy was found to decrease with increasing temperature, while that of liquid CMSX-10 superalloy increases with increasing temperature. The wetting behavior of liquid CM186LC on the alumina substrate was found (1) to differ significantly from that of CMSX-4 and CMSX-10 and (2) to vary with time. A HfO₂-rich layer was found in the contact area of CM186LC with the alumina substrate, which could lead to some uncertainty in the value obtained for the surface tension determined for CM186LC.     

Optimization by distributed control of reactors with decaying catalyst Part II: First-order catalyst deactivation

The problem of optimally choosing the temperature T(z, t) as a function of time t and position z in a tubular fixed bed chemical reactor, so as to minimize the total yield of product over a fixed time period for a reaction-deactivation system with a slow decaying catalyst has been formulated for the case where the rate of catalyst decay is linearly dependent upon activity. Several characteristics of extremal control policies which supplement the theoretical characterization obtained for the case of general reaction-deactivation kinetics using Sirazetdinov and Degtyarev's maximum principle are indicated. Results are that the supplementary properties of the extremal controls for linear catalyst deactivation kinetics may be used to advantage to reduce the computational dimensionality in synthesizing the control policies. Numerical calculations are presented to illustrate this fact in the case of initial catalyst activity profiles which are not uniform nor continuous along the reactor bed.     

The quantitative analysis of coal processing solvents by combined g.c.-m.s.

The application of combined g.c.-m.s. with automated data processing to the rapid analysis of coal solvents has been described. An analytical method, using commercially available data processing routines, has been developed for the automatic quantitative analysis of specific aromatic and hydroaromatic compounds in coal processing solvents. To demonstrate the use of this technique, raw anthracene oil was hydrogenated under different conditions and the resulting hydrogenated anthracene oils were analysed. The results showed correlations between the hydrogen content and the absolute concentrations of specific compounds in the samples.     

Design and synthesis of conformationally constrained cyclophilin inhibitors showing a cyclosporin-A phenotype in C. elegans

Cyclophilin A (CypA) is a member of the immunophilin family of proteins and receptor for the immunosuppressant drug cyclosporin A (CsA). Here we describe the design and synthesis of a new class of small-molecule inhibitors for CypA that are based upon a dimedone template. Electrospray mass spectrometry is utilised as an initial screen to quantify the protein affinity of the ligands. Active inhibitors and fluorescently labelled derivatives are then used as chemical probes for investigating the biological role of cyclophilins in the nematode Caenorhabditis elegans.