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Metallic bone plates are commonly used for arm bone fractures where conservative treatment (casts) cannot provide adequate support and compression at the fracture site. These plates, made of stainless steel or titanium alloys, tend to shield stress transfer at the fracture site and delay the bone healing rate. This study investigates the feasibility of adopting advanced composite materials to overcome stress shielding effects by optimising the geometry and mechanical properties of the plate to match more closely to the bone. An ulnar transverse fracture is characterised and finite element techniques are employed to investigate the feasibility of a composite-plated fractured bone construct over a stainless steel equivalent. Numerical models of intact and fractured bones are analysed and the mechanical behaviour is found to agree with experimental data. The mechanical properties are tailored to produce an optimised composite plate, offering a 25% reduction in length and a 70% reduction in mass. The optimised design may help to reduce stress shielding and increase bone healing rates.

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Completely resorbable composites are an attractive alternative for metallic bone-fracture fixation devices. However, failure of their interfacial integrity within aqueous environments, which can lead to a rapid loss of overall mechanical properties, has been reported in the literature. In this study coupling agents were investigated for phosphate glass fibre reinforced poly(lactic acid) composites. Three coupling agents with varying wettability were employed to improve initial mechanical properties and their retention in vitro via improvement of the interfacial bond between polymer matrix and fibres. Coupling agents were grafted onto the glass fibres by dip-coating in coupling agent solution at optimised concentrations. Three-aminopropyltriethoxy silane and sorbitol ended PLA oligomer treatments improved the initial flexural properties (27% strength with APS and 17% modulus via SPLA treatment) of the composites and 3-aminopropyltriethoxy silane and hexamethylene diisocyanate (HDI) treatments also decreased the loss of flexural strength and modulus during degradation. HDI treated samples retained 57.2% and 64.7% of their initial strength and modulus, respectively compared to control where only 34% of initial strength and 52% of initial modulus was retained after 28 days of degradation in PBS solution. Initial improvements in flexural properties were associated with improved shear bond strength at the interface due to covalent bonding between the glass fibres and polymer matrix provided by the coupling agents. Delay in mechanical property loss with degradation was suggested to be due to the hydrophobicity at the interface, which could have hindered the interfacial integrity loss and consequently loss of mechanical integrity of the composites. All coupling agent treated and control composites were tested for cytocompatibility using a primary human osteoblast cell line. A comparable response to the control, in terms of cell adhesion, proliferation and differentiation was observed supporting the use of these agents within implantable devices.

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Fully resorbable phosphate glass fiber reinforced polymer composites have shown real potential for replacing some of the existing metallic bone fracture fixation devices. However, some of these composites have not provided suitable mechanical strength profiles over the required healing period for bone. Typically, it has been seen that these composites can lose up to 50% or more of their strength within the first week of degradation. Functionalizing the glass surface to promote polymer adhesion or to introduce hydrophobicity at the glass surface could potentially introduce control over the mechanical properties of the composite and their retention. In this study eight chemical agents namely, Glycerol 2-phosphate disodium salt; 3-phosphonopropionic acid; 3-aminopropyltriethoxy silane; etidronic acid; hexamethylene diisocyanate; sorbitol/sodium ended PLA oligomers and amino phosphonic acid, were selected to functionalise the bulk phosphate glass surface. Selected chemical agents had one functional group (-OH or O C N) to react with the glass and another functionality (either -OH, NH2, or Na) to react with the polymer matrix and/or produce hydrophobicity at the fiber surface. Bulk phosphate glass surface-treated with the above agents were assessed for the cytotoxicity of degradation products cell-material interaction in short- and long-term direct cytocompatibility studies. Results obtained from these cytocompatibility studies (using human osteosarcoma (MG63) and primary human osteoblast cell lines) revealed no cytotoxicity from the degradation products and a response comparable to controls in terms of cell functions (attachment, viability, metabolic activity, proliferation, and differentiation) and morphology.

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Elucidation of the drivers of soil carbon (C) change is required to enable decisions to be made on how to achieve soil C sequestration. Interactions between different components in the ecosystem in combination with feedback mechanisms mean that identifying drivers through conventional experimental approaches or by retro-fitting models to data are unlikely to result in the insights needed for the future. This paper explains soil C dynamics by using a process-based model. Drivers considered in the model include nitrogen (N) fertiliser inputs, intake demand, and off-take of animal products. The effect of the grazing animal in uncoupling the C and N cycles is explained, plus the implications of the farming system ('drystock' versus milk). The model enables depiction of the dynamic equilibrium achieved with time when a proposed change in the drivers is sustained. The results show that soil C loss under lactating cows is a result of N, rather than C, being removed in milk. Counter-intuitively, at the same intake demand, N loss under 'milk' is less than under 'dry-stock', as is C loss in animal respiration. Possibilities for changing the longevity of C in the soil are discussed, and the compromise between food production, N loss and C sequestration is considered.

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Fibre reinforced composites have recently received much attention as potential bone fracture fixation applications. Bioresorbable composites based on poly lactic acid (PLA) and phosphate based glass fibre were investigated according to ion release, degradation, biocompatibility and mechanical retention profiles. The phosphate based glass fibres used in this study had the composition of 40P2O5-24MgO-16CaO-16Na2O-4Fe2O3 in mol% (P40). The degradation and ion release profiles for the composites showed similar trends with the amount of sodium and orthophosphate ions released being greater than the other cations and anions investigated. This was attributed to low Dietzal's field strength for the Na(+) in comparison with Mg(2+) and Ca(2+) and breakdown of longer chain polyphosphates into orthophosphate ions. P40 composites exhibited good biocompatibility to human mesenchymal stem cells (MSCs), which was suggested to be due to the low degradation rate of P40 fibres. After 63 days immersion in PBS at 37 °C, the P40 composite rods lost ~1.1% of mass. The wet flexural, shear and compressive strengths for P40 UD rods were ~70%, ~80% and ~50% of their initial dry values after 3 days of degradation, whereas the flexural modulus, shear and compressive strengths were ~70%, ~80%, and ~65% respectively. Subsequently, the mechanical properties remained stable for the duration of the study at 63 days. The initial decrease in mechanical properties was attributed to a combination of the plasticisation effect of water and degradation of the fibre-matrix interface, with the subsequent linear behaviour being attributed to the chemical durability of P40 fibres. P40 composite rods showed low degradation and ion release rates, good biocompatibility and maintained mechanical properties similar to cortical bone for the duration of the study. Therefore, P40 composite rods have huge potential as resorbable intramedullary nails or rods.

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Use of bioresorbable screws could eliminate disadvantages associated with metals such as removal operations, corrosion, MRI interference and stress shielding. Mechanical properties of bioresorbable polymers alone are insufficient for load bearing applications application as screws. Thus, reinforcement is necessary to try and match or surpass the mechanical properties of cortical bone. Phosphate based glass fibres were used to reinforce polylactic acid (PLA) in order to produce unidirectionally aligned (UD) and unidirectionally plus randomly distributed (UD/RM) composite screws (P40 UD and P40 UD/RM). The maximum flexural and push-out properties for the composite screws (P40 UD and P40 UD/RM) increased by almost 100% in comparison with the PLA screws. While the pull-out strength and stiffness of the headless composite screws were ∼80% (strength) and ∼130% (stiffness) higher than for PLA, those with heads exhibited properties lower than those for PLA alone as a result of failure at the heads. An increase in the maximum shear load and stiffness for the composite screws (∼30% and ∼40%) in comparison to the PLA screws was also seen. Maximum torque for the PLA screws was ∼1000 mN m, while that for the composite screws were slightly lower. The SEM micrographs for P40 UD and P40 UD/RM screws revealed small gaps around the fibres, which were suggested to be due to buckling of the UD fibres during the manufacturing process.

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Bioresorbable screws have the potential to overcome some of the complications associated with metallic screws currently in use. Removal of metallic screws after bone has healed is a serious issue which can lead to refracture due to the presence of screw holes. Poly lactic acid (PLA), fully 40 mol% P(2)O(5) containing phosphate unidirectional (P40UD) and a mixture of UD and short chopped strand random fibre mats (P40 70%UD/30%RM) composite screws were prepared via forging composite bars. Water uptake and mass loss for the composite screws manufactured increased significantly to ∼1.25% (P=0.0002) and ∼1.1% (P<0.0001), respectively, after 42 days of immersion in PBS at 37 °C. The initial maximum flexural load for P40 UD/RM and P40 UD composite screws was ∼60% (P=0.0047) and ∼100% (P=0.0037) higher than for the PLA screws (∼190 N), whilst the shear load was slightly higher in comparison to PLA (∼2.2 kN). The initial pull-out strengths for the P40 UD/RM and PLA screws were similar whereas that for P40 UD screws was ∼75% higher (P=0.022). Mechanical properties for the composite screws decreased initially after 3 days of immersion and this reduction was ascribed to the degradation of the fibre/matrix interface. After 3 days interval the mechanical properties (flexural, shear and pull-out) maintained their integrity for the duration of the study (at 42 days). This property retention was attributed to the chemical durability of the fibres used and stability of the matrix properties during the degradation process. It was also deemed necessary to enhance the fibre/matrix interface via use of a coupling agent in order to maintain the initial mechanical properties acquired for the required period of time. Lastly, it is also suggested that the degrading reinforcement fibres may have the potential to buffer any acidic products released from the PLA matrix.

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Six phosphate glass formulations (in the system P(2)O(5)-CaO-MgO-Na(2)O-Fe(2)O(3)) were produced with fixed magnesium and calcium content at 24 and 16 mol%, respectively. P(2)O(5) and Fe(2)O(3) were varied between 40-50 and 0-4 mol% respectively, with the balance being Na(2)O. EDX analyses confirmed the final composition of the glasses investigated to within a 1-2 % error margin. Thermal analyses showed a linear increase in T(g) with increasing Fe(2)O(3) and P(2)O(5) contents, with Fe(2)O(3) showing a greater effect than P(2)O(5). This was proposed to be due to the formation of Fe-O-P bonds and an increase in the cross-link density of the glass network enhancing the durability of the glass. The glasses that were investigated revealed a decrease in degradation rate with increasing Fe(2)O(3) and P(2)O(5) contents and again the effect of Fe(2)O(3) was greater. All the above characteristics correlated well with structural changes measured by IR and XPS analyses. Cytocompatibility studies showed good cellular (MG63) response to the glasses up to 168 h in terms of cell viability, proliferation and differentiation. Statistical analysis revealed that all the formulations with the exception of P50Fe4 gave a comparable response to the control (TCP), which suggested that after a threshold level of glass durability is achieved the degradation rate has no or minimal effect on biocompatibility. However, it was seen that the glass chemistry can also affect cellular response, since increasing the P(2)O(5) content promoted phenotypic expression that was not related to degradation rate but to the degradation products. This was supported using an elution assay.

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High-sugar grass varieties have received considerable attention for their potential ability to decrease N excretion in cattle. However, feeding high-sugar grasses alters the pattern of rumen fermentation, and no in vivo studies to date have examined this strategy with respect to another environmental pollutant: methane (CH(4)). Modeling allows us to examine potential outcomes of feeding strategies under controlled conditions, and can provide a useful framework for the development of future experiments. The purpose of the present study was to use a modeling approach to evaluate the effect of high-sugar grasses on simulated CH(4) emissions in dairy cattle. An extant dynamic, mechanistic model of enteric fermentation and intestinal digestion was used for this evaluation. A simulation database was constructed and analysis of model behavior was undertaken to simulate the effect of (1) level of water-soluble carbohydrate (WSC) increase in dietary dry matter, (2) change in crude protein (CP) and neutral detergent fiber (NDF) content of the plant with an increased WSC content, (3) level of N fertilization, and (4) presence or absence of grain feeding. Simulated CH(4) emissions tended to increase with increased WSC content when CH(4) was expressed as megajoules per day or percent of gross energy intake, but when CH(4) was expressed in terms of grams per kilogram of milk, results were much more variable due to the potential increase in milk yield. As a result, under certain conditions, CH(4) (g/kg of milk) decreased. The largest increases in CH(4) emissions (MJ/d or % gross energy intake) were generally seen when WSC increased at the expense of CP in the diet and this can largely be explained by the representation in the model of the type of volatile fatty acid produced. Effects were lower when WSC increased at the expense of NDF, and intermediary when WSC increased at the expense of a mixture of CP and NDF. When WSC increased at the expense of NDF, simulated milk yield increased and, therefore, CH(4) (g/kg of milk) tended to decrease. Diminished increases of CH(4) (% gross energy intake or g/kg of milk) were simulated when DMI was increased with elevated WSC content. Simulation results suggest that high WSC grass, as a strategy to mitigate N emission, may increase CH(4) emissions, but that results depend on the grass composition, DMI, and the units chosen to express CH(4). Overall, this project demonstrates the usefulness of modeling for hypothesis testing in the absence of observed experimental results.

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Several studies have investigated self-reinforced polylactic acid (SR-PLA) and polyglycolic acid (SR-PGA) rods which could be used as intramedullary (IM) fixation devices to align and stabilise bone fractures. This study investigated totally bioresorbable composite rods manufactured via compression moulding at ~100 °C using phosphate glass fibres (of composition 50P(2)O(5)-40CaO-5Na(2)O-5Fe(2)O(3) in mol%) to reinforce PLA with an approximate fibre volume fraction (v(f)) of 30%. Different fibre architectures (random and unidirectional) were investigated and pure PLA rods were used as control samples. The degradation profiles and retention of mechanical properties were investigated and PBS was selected as the degradation medium. Unidirectional (P50 UD) composite rods had 50% higher initial flexural strength as compared to PLA and 60% higher in comparison to the random mat (P50 RM) composite rods. Similar initial profiles for flexural modulus were also seen comparing the P50 UD and P50 RM rods. Higher shear strength properties were seen for P50 UD in comparison to P50 RM and PLA rods. However, shear stiffness values decreased rapidly (after a week) whereas the PLA remained approximately constant. For the compressive strength studies, P50 RM and PLA rods remained approximately constant, whilst for the P50 UD rods a significantly higher initial value was obtained, which decreased rapidly after 3 days immersion in PBS. However, the mechanical properties decreased after immersion in PBS as a result of the plasticisation effect of water within the composite and degradation of the fibres. The fibres within the random and unidirectional composite rods (P50 RM and P50 UD) degraded leaving behind microtubes as seen from the SEM micrographs (after 28 days degradation) which in turn created a porous structure within the rods. This was the main reason attributed for the increase seen in mass loss and water uptake for the composite rods (~17% and ~16%, respectively).

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Internal fixation for bone fractures with rigid metallic plates, screws and pins is a proven operative technique. However, refracture's have been observed after rigid internal fixation with metal plates and plate fixation has been known to cause localised osteopenia under and near the plate. In the present study, resorbable composites comprising a PLA matrix reinforced with iron doped phosphate glass fibres were investigated. Non-woven random mat laminates of approximately 30% and 45% fibre volume fraction (V(f)) were produced, along with unidirectional and 0°-90° samples of approximately 20% V(f). The non-woven composite laminates achieved maximum values of 10 GPa modulus and 120 MPa strength. The 0-90º samples showed unexpectedly low strengths close to matrix value (~50 MPa) although with a modulus of 7 GPa. The UD specimens exhibited values of 130 MPa and 11.5 GPa for strength and modulus respectively. All the modulus values observed were close to that expected from the rule of mixtures. Samples immersed in deionised water at 37°C revealed rapid mechanical property loss, more so for the UD and 0-90º samples. It was suggested that continuous fibres wicked the degradation media into the composite plates which sped up the deterioration of the fibre-matrix interface. The effect was less pronounced in the non-woven random mat laminates due to the discontinuous arrangement of fibres within the composite, making it less prone to wicking. Random mat composites revealed a higher mass loss than the UD and 0°-90° specimens, it was suggested this was due to the higher fibre volume fractions of these composites and SEM studies revealed voidage around the fibres by day 3. Studies of pH of the degradation media showed similar profiles for all the composites investigated. An initial decrease in pH was attributed to the release of phosphate ions into solution followed by a gradual return back to neutral.

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High-sugar grass varieties have received considerable attention for their potential to reduce nitrogen (N) excretion and increase milk yield in cattle. However, considerable variation exists in the magnitude of response in published results. The purpose of this study is to explain the variation in response using a dynamic mechanistic model to predict observed N and milk yield results from the literature, and from simulated data. Examined effects were (1) water-soluble carbohydrate [WSC; g/kg of dry matter (DM)] increase; (2) change in crude protein (CP) and neutral detergent fiber (NDF) content of the plant with WSC increase; and (3) the level of N fertilization. The database for evaluation of model N and milk yield predictions consisted of 4 published studies with 28 treatment means for which high-sugar grasses were being evaluated. Water-soluble carbohydrate content of the diets ranged from 95 to 248 g/kg of DM, CP content ranged from 115 to 263 g/kg of DM, and the NDF content ranged from 400 to 568 g/kg of DM. Urine N, milk N, and total N excretion were predicted well by the model and followed the directional pattern of observed values within each study. Simulation results showed that the N utilization ratio increased as the WSC content of the diet increased, but to varying degrees depending on the grass scenario examined. The greatest benefit in terms of N utilization ratio and urine N levels were seen when the WSC content of grass increased at the expense of CP, followed by a 50:50 CP and NDF mix, followed by a trade for NDF. Simulated milk yield decreased slightly when WSC increased at the expense of CP, increased slightly when it increased at the expense of a CP and NDF mix, and increased most when WSC increased at the expense of NDF. Results were amplified slightly under conditions of low-N fertilization and in the absence of grain feeding. Overall, modeling is useful as an explanatory tool. The variation from results in the literature with high-WSC grass feeding may be, at least in part, the result of the level of WSC (g/kg of DM) increase, concurrent changes occurring within the CP and NDF fractions of the plant, and the plane of nutrition of the diet (grain feeding and N fertilization levels).

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We report results from an initial small animal study designed to provide information on the biocompatibility of a novel biodegradable composite designed for craniomaxillofacial reconstruction. Rat calvarium was chosen as a clinically analogous model, which allowed comparison between experimental groups (PCL alone, PCL/phosphate glass, or PCL/bioglass implants) and control groups (empty defects or bone grafted defects). All animals recovered well from surgery and no clinical complications were observed. Histological assessment indicated a lack of inflammatory response. The amount of new bone formation at the dural aspect of the implant was statistically significantly higher in the PCL/phosphate glass group than the other experimental groups. This study confirms, in a clinically analogous model, the promise of the novel PCL/phosphate glass composite material. Work is planned toward manufacturing scale up and clinical trials.

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Details of the microscopic structure of phosphate glasses destined for biomedical applications, which include sodium, magnesium and calcium cations, have been obtained from the static structure factor measured by means of neutron scattering. A complementary, molecular dynamics study has been performed on a range of phosphate glasses using density functional theory methods, which allow structural fluctuations, including bond breaking, in the liquid phase before quenching to the glass phase. Good agreement between experiment and simulation allows the molecular dynamics trajectories to be analysed in detail. In particular, attention is focused on the cross-linking of divalent cations in contrast with the structural aspects associated with monovalent cations. Magnesium cations are found equidistant and bridging between the phosphorus atoms of different phosphate chains, leading to a shorter phosphorus-phosphorus second neighbour distance (that is, a more compact packing of neighbouring phosphate chains) compared to the effect of sodium cations. Calcium cations show behaviour intermediate between those of magnesium and sodium. Molecular dynamics simulations give access to the cation mobility, which is lowest for magnesium, reflecting its structural, cross-linking role.

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Polymers prepared from polylactic acid (PLA) have found a multitude of uses as medical devices. The main advantage of having a material that degrades is so that an implant would not necessitate a second surgical event for removal. In addition, the biodegradation may offer other advantages. In this study, fibers produced from a quaternary phosphate-based glass (PBG) in the system 50P(2)O(5)-40CaO-5Na(2)O-5Fe(2)O(3) (nontreated and heat-treated) were used to reinforce the biodegradable polymer, PLA. Fiber properties were investigated, along with the mechanical and degradation properties and cytocompatibility of the composites produced. Retention of mechanical properties overtime was also evaluated. The mean fiber strength for the phosphate glass fibers was 456 MPa with a modulus value of 51.5 GPa. Weibull analysis revealed a shape and scale parameter value of 3.37 and 508, respectively. The flexural strength of the composites matched that for cortical bone; however, the modulus values were lower than those required for cortical bone. After 6 weeks of degradation in deionized water, 50% of the strength values obtained was maintained. The composite degradation properties revealed a 14% mass loss for the nontreated and a 10% mass loss for the heat-treated fiber composites. It was also seen that by heat-treating the fibers, chemical and physical degradation occurred much slower. The pH profiles also revealed that nontreated fibers degraded quicker, thus correlating well with the degradation profiles. The in vitro cell culture experiments revealed both PLA (alone) and the heat-treated fiber composites maintained higher cell viability as compared to the nontreated fiber composites. This was attributed to the slower degradation release profiles of the heat-treated composites as compared to the nontreated fiber composites. SEM analyses revealed a porous structure after degradation, and it is clear that there are possibilities here to tailor the distribution of porosity within polymer matrices.

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Composites comprising a biodegradable polymeric matrix and a bioactive filler show considerable promise in the field of regenerative medicine, and could potentially serve as degradable bone fracture fixation devices, depending on the properties obtained. Therefore, glass fibres from a binary calcium phosphate (50P(2)O(5)+50CaO) glass were used to reinforce polycaprolactone, at two different volume fractions (V(f)). As-drawn, non-treated and heat-treated fibres were assessed. Weight loss, ion release and the initial mechanical properties of the fibres and composites produced have been investigated. Single fibre tensile testing revealed a fibre strength of 474MPa and a tensile modulus of 44GPa. Weibull analysis suggested a scale value of 524. The composites yielded flexural strength and modulus of up to 30MPa and 2.5GPa, respectively. These values are comparable with human trabecular bone. An 8% mass loss was seen for the lower V(f) composite, whereas for the two higher V(f) composites an approximate 20% mass loss was observed over the course of the 5week study. A plateau in the degradation profile at 350h indicated that fibre dissolution was complete at this interval. This assertion was further supported via ion release studies. The leaching of fibres from the composite created a porous structure, including continuous channels within the polymer matrix. This offers further scope for tailoring scaffold development, as cells from the surrounding tissue may be induced to migrate into the resulting porous matrix.

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Changes in the thermal properties of sodium phosphate glasses during melt production have been investigated using Pt/Au and fused alumina crucibles. Glasses were produced from NaH(2)PO(4) as a starting material, providing an intrinsic Na(2)O:P(2)O(5) ratio of 1:1 and giving an O/P = 3, that is, a metaphosphate. In Pt/Au crucibles, glass transition temperatures rose to a plateau value of 295 degrees C at a rate determined by melt temperature. No contamination of the glass by platinum or gold was detected or indicated in the results. E(a) for the reaction was found to be 66.4 kJ mol(-1). In fused alumina crucibles, glass transition temperatures rose to over 450 degrees C, with these values showing some convergence at higher furnace temperatures. Extensive erosion of the alumina crucibles was observed. The amount of alumina incorporation within the glasses correlated well with the rise in glass transition temperature up to a maximum of 15.5 mol % Al(2)O(3) content. Al(2)O(3) incorporation above this value caused a reduction in the value of the T(g).

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The degradation profiles of six sodium iron phosphate glass formulations have been investigated using simple dissolution trials in deionized water. The glasses were produced from the appropriate phosphate salts by melting at 1200 degrees C in 5% Au/95% Pt crucibles. Dissolution rates varied from 0.2 gcm(-2)h(-1) for the 1% Fe glass to essentially zero over the 6-week test period for the 15% Fe and 20% Fe glasses. The overall degradation rate was found to vary according to the approximate relation: rate = 1.3e(-0.79x) gcm(-2)h(-1), where x is the percentage iron content of the glass. Glasses with 10% or greater iron content were observed to maintain a constant density over the course of the tests and thus appeared to degrade from the surface and not the bulk. In vitro cell response tests were conducted on the glasses using macrophages and primary craniofacial osteoblasts. These tests were performed on the glasses with 10% or greater iron content because glasses with lower iron content degraded too quickly. Confocal microscopy revealed a rounded macrophage morphology and IL-1beta production was low, suggesting little macrophage activation. However, a significant level of peroxide production was observed. Osteoblasts were observed to attach to the glass surfaces and spread, exhibiting a similar cytosketetal organization to the cells on the Thermanox controls, with a high level of F-actin organization. On balance, the 15% Fe glass performed slightly better than the 20% Fe glass in these assays.

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Experiments investigating the population responses of aphids to CO2 enrichment have yielded results suggesting that aphid populations will be both larger under elevated CO2 and that they will be smaller under elevated CO2. Most studies have failed to reject the null hypothesis of no difference in population sizes due to atmospheric CO2 concentration. This diversity of results has led some investigators to conclude that aphid responses are not general, and that every aphid-plant interaction may be unique and unpredictable a priori. We use a single, general, mathematical model to consider the population responses of cereal aphids to grass grown under different CO2 concentrations. The model shows that it is possible to explain any of the three observed results: larger populations, smaller populations, or no difference, and that which of these three outcomes arises may depend critically on the interaction between aphid nitrogen requirements and the nitrogen fertility of the soil. The model also shows that the qualitative results will depend on how sensitive the aphid species is to increases in its own density. Past studies have shown that aphids increase their production of winged offspring in response to increasing aphid density. The model predicts that, in general, aphid species that have lower nitrogen requirements and that are less sensitive to their own density will be more likely to have larger populations in elevated CO2 compared to ambient CO2. Differences between aphid species (and clones) in their nitrogen requirements and the strength of their density-dependent response have not been widely reported in the literature. Also, the nitrogen fertility of the soil has rarely been manipulated in experiments on aphid responses to rising CO2 levels. The model suggests that the diversity of population responses of aphids may be both understandable and predictable in the context of such an interaction.

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BACKGROUND AND PURPOSE: DNA ploidy and cell cycle measurements of uveal melanoma tissue are regarded as having limited prognostic significance. In contrast, dual-parameter (DNA monoclonal antibody) flow cytometry offers a convenient and rapid way to screen tumour samples for a variety of phenotypic markers, whilst simultaneously measuring DNA ploidy and cell cycle, and therefore has the increased potential to identify clinically relevant indicators of disease progression. The aim of the present study was to identify a simple yet robust method for isolating, preserving and staining cells that could be analysed by flow cytometry. METHODS: Using a simple preparation procedure, a panel of membrane-associated antibodies (ICAM-1, W632, HLA-DR) and nuclear or cytoplasmic oncoprotein antibodies (c-erbB-2, c-myc, bcl-2, p53), together with positive (PHM-5) and negative (FITC F(ab')2) controls, were assayed. It was considered important to test the protocol with markers expressed on the cell surface, and in the cytoplasm and nucleus, so as not to be restrictive and thereby exclude an antigen of potential clinical interest. In addition, such panels would also enable the generation of a 'phenotypic profile' for each specimen that may reveal clinically significant trends. RESULTS: Our results indicate that tissue dissociation followed by brief fixation in 1% paraformaldehyde and permeabilisation in 70% methanol produces a stable single cell suspension, which can subsequently be stained with a wide range of antibodies for the accurate identification of cells in a potentially heterogeneous tumour population. CONCLUSION: This technology can rapidly identify sub-populations of cells expressing differing levels of proteins, which may prove to be indicative of disease progression for this aggressive disease.

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