In nature, the majority of bacteria grow as biofilms in mixed consortia that use mutualistic, syntrophic, commensal or antagonistic strategies to compete for and efficiently utilize available nutrients [1–4]. Microbial biofilms are critically important in medical, environmental and engineered biological systems. For example, the human gut microbiome has emerged as a major focus for biomedical research with mounting evidence suggesting unhealthy gut flora biofilms are associated with illnesses including autoimmune diseases, colorectal cancer and inflammatory bowel disease [5–10]. Environmental microbial biofilm consortia form the basis of global nutrient cycles from nitrogen fixation to carbon fluxes [11, 12]. Additionally, the study of natural biofilms has gained in popularity recently due to their efficient organization and ability, through synergistic interactions, to optimize multiple tasks simultaneously like the deconstruction of complex, recalcitrant plant materials into simple sugars. A major goal of current biofuels research is to engineer synthetic microbial communities that mimic these naturally occurring biofilms for biomass conversion to renewable liquid fuels . While foundational to the vast majority of microbial life on the planet, the basic design principles of consortial biofilms are still poorly understood due largely to the complexity of naturally occurring systems [3, 4].
Chronic wounds are defined as a host-pathogen environment that has failed to proceed through a timely healing process. An estimated 2 % of the U.S. population (6 million people) have a non-healing chronic wound with treatment costing more than $25 billion per year [14–16]. Chronic wounds are often colonized by microorganisms growing as biofilms on a complex mixture of wound exudate [17–24]. Microbes residing in biofilms exhibit phenotypes distinct from planktonic growth, making treatment a major challenge. For instance, bacteria in biofilms can tolerate antimicrobial agent concentrations 10,000 times higher than the same microbes grown planktonically [25, 26]. Chronic wounds are typically colonized by consortia comprised of different microbial species [17–19, 27, 28]. Polymicrobial infections have been reported to have elevated mortality rates relative to monocultures , and in vivo rabbit model systems demonstrated that consortia prevented wound healing compared to their respective monocultures [24, 30].
The aerobe Pseudomonas aeruginosa and the facultative anaerobe Staphylococcus aureus are two bacteria commonly isolated from chronic would biofilm infections [19, 23, 27]. The same two bacteria are often key contributors to multispecies infections that occur in the lung mucous of cystic fibrosis patients . P. aeruginosa is known to exhibit much lower growth rates than S. aureus and other facultative anaerobes in anaerobic environments common in chronic wound and mucoid biofilms [32, 33]. Perhaps partially in response to this metabolic disadvantage, P. aeruginosa has evolved a number of mechanisms to enhance its competitiveness in multispecies biofilm communities. The most widely studied mechanism is growth inhibition and lysis of competing bacteria through the secretion of an arsenal of small molecule (e.g. pyocyanin ) and protein (e.g. bacteriocins ) toxins. The consumption of metabolic byproducts secreted by other bacteria through cross feeding mechanisms also has been proposed to enhance P. aeruginosa competitiveness . Another putative mechanism is P. aeruginosa chemotaxis towards high oxygen niches (i.e. aerotaxis ) where it is metabolically competitive.
Multispecies biofilms are sufficiently complex to preclude detailed understanding through traditional experimental techniques developed for planktonic cultures. A primary challenge is the complex interactions between the biofilm species and the extracellular environment . Most naturally occurring microbial consortia exist in spatially heterogeneous environments that also exhibit temporal variations. The presence of spatial heterogeneity plays an essential role in the evolution and function of microbial species [39–43] and has profound effects on biofilm formation and development [3, 38, 44, 45]. Concentration gradients in key nutrients due to limited diffusion establish metabolic niches within the biofilm that can produce spatial variations in biomass density  and additionally spatial partitioning of species in the case of multispecies biofilms [45, 47]. Quantitative understanding of the relationships between spatial and temporal variations in the extracellular environment and community metabolism is critical to systematically analyze and rationally manipulate biofilm consortia. While spatiotemporal metabolic models that account for both spatial and temporal variations in the extracellular environment have been constructed, these models rely on table lookups of precomputed flux balance solutions [48–50] or lattice based descriptions of nutrient diffusion [51, 52].
We recently proposed a general methodology for spatiotemporal metabolic modeling based on combining genome-scale reconstructions with fundamental transport equations that capture the relevant convective  and/or diffusional  processes. In this paper, we applied this methodology to develop biofilm metabolic models that predict the complex spatiotemporal behavior of a P. aeruginosa/S. aureus two species system. The biofilm models were formulated by combining genome-scale reconstructions of P. aeruginosa and/or S. aureus metabolism with uptake kinetics and reaction-diffusion type equations for extracellular substrates and metabolic byproducts. To avoid complications associated with solving a moving boundary problem, the biofilm was assumed to have a fixed thickness over which metabolites diffused and cell growth occurred. Therefore, the models were most appropriate for predicting the metabolism of biofilms of a specified thickness. We developed an effective computational method for solving the biofilm models, which consisted of a set of partial differential equations with mixed boundary conditions constrained by embedded linear programs. The models were used to analyze the metabolic differences between single species and two species chronic wound biofilms and to investigate putative factors that could impact the physiology of the two species biofilm, including nutrient diffusion, metabolite cross-feeding, P. aeruginosa motility and P. aeruginosa mediated lysis of S. aureus.
Biofilm models were formulated by combining genome-scale reconstructions of P. aeruginosa and/or S. aureus metabolism with uptake kinetics for available nutrients and reaction-diffusion type equations for species biomass, supplied substrates and synthesized metabolic byproducts. Single species biofilm models were formulated with either the P. aeruginosa or S. aureus reconstruction, while the two species model used both reconstructions. Diffusion was assumed to occur only in the axial direction of the biofilm such that spatial variations could be captured with a single variable z (Fig. 1a). For simplicity, the biofilm was assumed to have a fixed thickness W over which the nutrients diffused and cell growth occurred. Therefore, the models were most appropriate for predicting the metabolism of biofilms of a specified thickness. Both strains were assumed to consume glucose as the primary carbon source . Glucose was supplied at the tissue-biofilm interface at the assumed concentration of the wound exudate, while oxygen was supplied at the biofilm-air interface at a concentration for an aqueous solution in equilibrium with atmospheric oxygen.
The P. aeruginosa PA01 iMO1056 reconstruction accounts for 1056 genes, 1030 enzymes, 833 intracellular reactions and 277 exchange reactions . This reconstruction has been shown to provide good agreement with experimentally determined biomass yields for aerobic growth on glucose and anaerobic growth on glucose with nitrate as an electron acceptor. Our preliminary flux balance calculations with a maximum growth objective showed the primary metabolic byproducts to be acetate and L-alanine. The secretion fluxes of other minor byproducts were approximately an order of magnitude less than for acetate and L-alanine. P. aeruginosa is known to secrete acetate, lactate and succinate , while the secretion of L-alanine has not been reported. To obtain byproduct distributions in better agreement with , we constrained the L-alanine secretion flux to zero. This modification resulted in a redirection of flux from L-alanine to succinate with little effect on the secretion fluxes of acetate and minor byproducts. Furthermore, we enforced a minimal non-growth associated ATP maintenance flux of 5 mmol/gDW/h, the same value as in the S. aureus reconstruction, to reduce the P. aeruginosa anaerobic growth rate for consistency with experimental studies . The iMO1056 reconstruction contained succinate, lactate and acetate uptake fluxes that allowed the investigation of putative cross feeding of metabolic byproducts. Secretion of the small molecule inhibitor pycoyanin was included by adding an exchange flux with an adjustable lower bound that forced pycoyanin synthesis, which was in opposition to growth rate maximization.
The S. aureus N315 iMH551 reconstruction accounts for 551 genes, 604 enzymes, 682 intracellular reactions and 92 exchange reactions . This model correctly reproduces byproduct secretion patterns under aerobic conditions with glucose limitation and under anaerobic conditions with glucose excess [59–63]. Our flux calculations showed that the primary byproducts were acetate and lactate. The iMH551 reconstruction contained lactate and acetate uptake fluxes that allowed reassimilation of secreted metabolic byproducts. To explore the possibility of succinate cross feeding, the S. aureus model was modified to allow succinate uptake through a putative proton dependent symport mechanism.
where v i is the uptake rate (mmol/gDW/h) of the i-th substrate, S i is the extracellular concentration (mmol//L) of the i-th substrate, v max,i is the maximum uptake rate and K m,i is the half saturation constant. Equation (1) was used to establish transport bounds on the uptake rates with the actual uptake rates being determined by solution of the intracellular flux balance problem. Both v max,i and K m,i were important parameters due to the large nutrient spatial gradients induced by diffusion through the biofilm.
where X and Y are the biomass concentrations (g/L) of P. aeruginosa and S. aureus, respectively, Z = X + Y is the total biomass concentration, and μ X and μ Y are the corresponding growth rates (h−1) obtained from the flux balance calculations. If local nutrient concentrations became too small to meet the ATP maintenance demand of a species, then the flux balance problem for the species became infeasible at that location. Once an infeasibility was detected, the death rate constant k dX or k dY was rapidly increased exponentially from zero to a fixed, non-zero value to simulate that the species would begin to die at that location. This approach ensured that the model equations remained smooth and could be integrated. P. aeruginosa has flagella for motility and can aerotaxis towards higher oxygen levels . This capability was captured in the model by including a typical chemotaxis term  where O is the oxygen concentration (mmol/L) and k A is the aerotaxis rate constant. Cell growth was restricted to a maximum cell concentration Z max to account for cell crowding effects within the biofilm. No flux boundary conditions were imposed at the tissue-biofilm (z = 0) and biofilm-air (z = W) interfaces under the assumption that cells could not leave the biofilm via mechanisms such as dispersal.
where v P is the specific pyocyanin synthesis rate obtained from the flux balance calculation and D P is the pyocyanin diffusion coefficient. A no flux boundary condition was imposed at the biofilm-air interface assuming that the pyocyanin was non-volatile. By contrast, a Robin type boundary condition was imposed at the tissue-biofilm interface to describe possibly mass transfer limited removal of pyocyanin, where k mP is pyocyanin mass transfer coefficient and P b is the bulk pyocyanin concentration in the tissue.
where G is the glucose concentration (mmol/L), the P. aeruginosa uptake fluxes v GX and v OX and the S. aureus uptake fluxes v GY and v OY were obtained from the flux balance calculations, and D G and D O are the glucose and oxygen diffusion coefficients. For glucose, a no flux boundary condition was imposed at the biofilm-air interface assuming glucose was not volatile and a Robin type boundary condition was imposed at the tissue-biofilm interface to model possibly mass transfer limited transport of glucose into the biofilm. Here k mG is the glucose mass transfer coefficient and G b is the bulk glucose concentration in the wound exudate. For oxygen, Robin type boundary conditions were imposed at both interfaces with oxygen mass transfer coefficient k mO , oxygen concentration O b at the tissue-biofilm interface and oxygen concentration O a at the biofilm-air interface.
where M j is concentration (mmol/L) of the j-th byproduct (A = acetate, S = succinate, L = lactate), the secretion (or consumption) fluxes v MjX and v MjY were obtained from the flux balance calculations, D Mj is the diffusion coefficient, k mMj is the mass transfer coefficient at the tissue-biofilm interface, and M j,b is the bulk concentration of the wound exudate. No flux boundary conditions were imposed at the biofilm-air interface assuming the byproducts were not volatile, while a Robin type boundary condition was imposed at the tissue-biofilm interface to allow removal of the byproducts from the biofilm.
where D Mk is the biofilm diffusion coefficient of the k-th species (glucose, oxygen, acetate, succinate, lactate and pyocyanin), D WMk is the aqueous diffusion coefficients at 37 °C and a MK is an adjustable parameter. These relations were obtained using a previously proposed empirical equation  by adjusting a MK such that the biofilm diffusion coefficient was equal to the aqueous value when Z = 0 and equal to biofilm values reported elsewhere  when Z = Z max. Because pyocyanin has a comparable molecular weight to glucose, the inhibitor diffusion coefficient was chosen to follow the same relation.
We found a dearth of literature for determining species specific values for the 20 parameters needed to calculate uptake rates with respect to the five possible nutrients (glucose, oxygen, succinate, lactate, acetate). Consequently, the two species were assumed to have the same uptake parameter values. We used representative glucose [68, 69] and oxygen [69, 70] uptake parameter values reported for the model bacterium Escherichia coli under the assumption that P. aeruginosa and S. aureus should have similar values. Because we were not able to find reliable uptake parameter values for succinate, lactate and acetate, the associated v max and K m values were assumed to be equal to those for glucose (Table 1). Therefore results focused on differences in metabolic network structure of the two species and not on differences in uptake properties.
Other parameter values for the biofilm model were obtained from the literature to the extent possible (Table 2). We utilized a typical biofilm thickness W and assumed wound exudate concentrations consistent with published values. The air oxygen concentration O a was derived from the oxygen content of atmospheric air. P. aeruginosa and S. aureus cell death were implemented by exponentially increasing the death rate constants from zero to the values listed in Table 2 when local nutrient concentrations were not sufficient to meet ATP maintenance demands. The lower bound on the P. aeruginosa pyocyanin synthesis flux v P,min was tuned such that the average pyocyanin concentration within the biofilm was the same order of magnitude as that observed experimentally in . The S. aureus inhibitor-mediated death constant k L was tuned to achieve reasonable spatial distributions of the two species, which included P. aeruginosa dominance in the aerobic region of the biofilm, S. aureus dominance in the anaerobic region and a sharp spatial division between the two species [47, 72–74].
The maximum achievable biomass concentration Z max was chosen to be within the large range of published values . We established reasonable metabolite concentrations within the biofilm by adjusting a single mass transfer coefficient for glucose, acetate, succinate, lactate and pyocyanin such that their average concentrations were the same order of magnitude as those observed experimentally in  and . The P. aeruginosa aerotaxis rate constant k A was chosen such that P. aeruginosa was dominant in the aerobic region of the biofilm and a sharp spatial division between the two species was established as the biofilm matured towards a steady-state condition [43, 47, 76]. Initial conditions for each simulation were generated by first running a simulation with each species biomass concentration constrained to be 1 g/L and capturing the resulting steady-state solution. These initial conditions reflected a newly developed, nearly spatially homogeneous biofilm with small cell densities, high nutrient levels and low byproduct concentrations.
The two species biofilm model consisted of a set of partial differential equations (PDEs) with mixed boundary conditions and embedded linear programs (LPs). The efficient and stable solution of such models is a challenging problem at the forefront of microbial metabolic modeling . As described in our previous publications [53, 54], we pursued a spatial discretization approach based on converting the PDEs into a large set of ordinary differential equations (ODEs) in time with embedded LPs (Fig. 1b). The spatial domain [0, W] was discretized using N = 50 node points at which the diffusion terms in Eqs. (2) – (5) were discretized using central difference approximations with second-order accuracy. The specified boundary conditions were incorporated into the central difference approximations at the boundary node points. This procedure yielded a set of 8 ODEs at each node point for the local concentrations of P. aeruginosa and S. aureus biomass, glucose, oxygen, acetate, succinate, lactate and pyocyanin.
This ODE system was solved using DFBAlab , a MATLAB tool that explicitly addresses problems associated with LP alternative optima and possible infeasibilities . DFBAlab employs a lexicographic optimization strategy in which a series of LP problems are sequentially solved to ensure the determination of unique exchange fluxes necessary for a well-defined dynamic system. Each LP is solved subject to constraints that the previous objectives are equal to their optimal values, with the required number of LPs equal to the number of exchange fluxes. We specified the lexicographic optimization objectives to reflect known or anticipated physiology of the two species biofilm community (Table 3). We found that reordering these objectives had no noticeable effect on simulation results. Each node point was represented by 8 ODEs for the local species and metabolite concentrations and 12 LPs for lexicographic optimization. We employed 50 node points such that the discretized biofilm model consisted of 400 ODEs and 600 LPs.
All simulations were performed with MATLAB 8.5 (R2015a) using DFAlab, the stiff MATLAB integrator ode15s for dynamic flux balance model solution and Gurobi 6.0 for linear program solution. A typical 1000-h dynamic simulation for determining a steady-state solution required about 25 min running on an ASUS computer with Intel Core i7-960 processor and 24 GB RAM. As compared to alternative computational methods for spatiotemporal metabolic modeling based on table lookups of precomputed FBA solutions combined with integration of the PDEs on a coarse spatial grid [48–50] and real-time FBA solution combined with lattice-based descriptions of metabolite diffusion [51, 52], we believe our approach offers several important advantages including the use of DFBAlab, the ability to directly embed LPs within the discretized ODEs, and the flexibility to solve the ODE-LP system using stiff integrators with variable step size and error control.
Flux balance analysis (FBA) was performed on the single species, genome-scale reconstructions to investigate P. aeruginosa and S. aureus growth rates in different metabolic niches consistent with the bottom, middle and top of the biofilms. More specifically, FBA was used to predict growth rates at three different glucose and oxygen uptake rates. We also performed FBA at three different acetate and lactate uptake rates to investigate the effect of byproduct uptake on growth rates. The growth rate of S. aureus was predicted to equal or exceed that of P. aeruginosa for all combinations of glucose and oxygen uptake rates, with the difference most pronounced under anaerobic conditions expected at the bottom of the biofilm (Table 4). No growth was predicted for either species under anaerobic, low glucose conditions which may prevail in mature biofilms of sufficient thickness. When acetate was used as the carbon source, P. aeruginosa was predicted to have low growth rates that depended only on the acetate uptake rate until oxygen became limiting. S. aureus was not able to grow on acetate at any oxygenation level, as has been observed experimentally . As compared to glucose, the absolute growth rates and differences between species growth rates was relatively small for growth on lactate. Collectively, these results suggest that S. aureus has a distinct growth rate advantage over P. aeruginosa for all conditions anticipated in the simulated two species biofilm.
Dynamic simulations were performed for single species biofilms consisting of only P. aeruginosa or S. aureus with glucose and oxygen as the only substrates. Simulations were performed over a time window of 1000 h to capture a steady-state solution, although near steady-state behavior was typically established in <100 h. For each species, biofilms of different thicknesses were simulated to determine the maximum thickness W max that could be sustained under the given environmental conditions. If the biomass concentration dropped below 10 g/L (5 % of the maximum value Z max = 200 g/L) anywhere in the mature biofilm obtained after 1000 h of simulation, the thickness was deemed too large and reduced. These simulations predicted that S. aureus could grow much thicker biofilms with W max = 90 μm compared to P. aeruginosa with W max = 30 μm.
Spatially resolved predictions obtained after 1000 h showed that the P. aeruginosa biofilm (Fig. 2a) were characterized by high biomass concentrations throughout the biofilm, low oxygen concentrations near the bottom of the biofilm furthest from the oxygen source, low glucose concentrations near the top of the biofilm furthest from the glucose source, and acetate and succinate as the primary metabolic byproducts. If the biofilm thickness was chosen larger than W max = 30 μm, the P. aeruginosa biomass concentration dropped below 10 g/L at the bottom of the biofilm due to the combination of low oxygen penetration and very small anaerobic growth rates (see glucose results in Table 4). The S. aureus biofilm model predicted much deeper oxygen penetration (~50 μm vs. ~25 μm) due to more efficient use of oxygen for glucose oxidation (Fig. 2b). Superior anaerobic growth allowed S. aureus to produce much thicker biofilms under the same environmental conditions. Otherwise, the predictions were similar to those obtained with P. aeruginosa with lactate replacing succinate as a primary byproduct. If the S. aureus biofilm thickness was chosen larger than W max = 90 μm, the biomass concentration dropped below 10 g/L near the top of the biofilm due to inadequate glucose penetration. The effect of the metabolite mass transfer coefficients on concentration gradients within the S. aureus biofilm are shown in Additional file 1: Figure S1.
If the two species biofilm thickness was chosen larger than W max = 80 μm, the biomass concentration dropped below 10 g/L near the top of the biofilm due to inadequate glucose penetration. The relationship between the two species was interpreted to be antagonistic with the incorporation of P. aeruginosa into a S. aureus biofilm reducing the achievable thickness for a given set of conditions.
When the two species biofilm thickness was set equal to W max = 80 μm, pseudo steady-state solutions were obtained after only 50 h of simulation (Fig. 3a). Oxygen was quickly depleted throughout most of the biofilm, except near the biofilm-air interface where an aerobic region was established as observed experimentally . Similarly, glucose was rapidly depleted in all regions except near the tissue-biofilm interface where a glucose rich region was maintained. S. aureus was predicted to quickly establish dominance throughout the biofilm due to its higher local growth rates, especially in the anaerobic region. Initially acetate and succinate levels increased but thereafter they were predicted to decrease due to metabolite removal at the tissue-biofilm boundary. Lactate levels were predicted to remain high throughout the biofilm due to S. aureus synthesis in the anaerobic region and diffusion into the aerobic region. As in the single species biofilms, multispecies biofilm spatial profiles obtained after 1000 h of simulation (Fig. 3b) were characterized by the presence of a glucose rich, anaerobic region near the tissue-biofilm interface and a glucose depleted, aerobic region near the biofilm-air interface. S. aureus was predicted to be dominant throughout the biofilm, especially in the anaerobic region, while P. aeruginosa was predicted to be present only in the aerobic region. Byproduct profiles were similar to those obtained for the S. aureus single species biofilm (see Fig. 2b) with high lactate levels, low acetate levels and no succinate production. We attributed this behavior to partitioning of P. aeruginosa to the aerobic region where the synthesis of byproducts was substantially reduced.
To further analyze how multispecies metabolism depended on position in the biofilm, local effective growth rates and nutrient uptake rates were determined from the base case (BC) simulation data. For species i, the local effective growth rate was calculated as the difference between the biomass restricted growth rate μ i (1-Z i /Z max) and the energy associated death rate k di at a given position z. Consequently, the effective growth rate could be negative in nutrient lean regions. The calculations were performed using data collected at t = 10 h because these initial rates offered insights into biofilm physiology. S. aureus growth rates exceeded P. aeruginosa growth rates at all positions, especially in the anaerobic region near the bottom of the biofilm where P. aeruginosa death was predicted (Fig. 3c). Both species were predicted to have constant growth rates in the upper aerobic region. The P. aeruginosa growth rate decreased rapidly in the lower half due to decreasing oxygen availability such that death occurred in the first 35 μm. By contrast, the S. aureus growth rate increased rapidly in this region due to the increasing availability of glucose to support anaerobic growth. As time progressed, these local growth rates resulted in S. aureus dominance throughout the biofilm and P. aeruginosa presence only in the aerobic region (Fig. 4a). The glucose uptake rate increased monotonically from bottom to top of the biofilm, while the oxygen uptake rate was predicted to exhibit a maximum near the center because that location offered the optimal combined availability of glucose and oxygen to support consortium growth.
We also performed simulations assuming additional oxygen was available at the biofilm-tissue interface at a blood plasma concentration of 0.05 mmol/L . This oxygen source allowed P. aeruginosa growth near the biofilm-tissue interface and therefore reduced spatial partitioning of the two species (Additional file 2: Figure S2). However, S. aureus still dominated throughout the biofilm due to it higher aerobic and anaerobic growth rates.
We hypothesized that cross feeding of secreted metabolic byproducts (lactate, succinate, acetate) would enhance the competitiveness of P. aeruginosa in the aerobic portion of the biofilm. Except for the inability of S. aureus to consume acetate, experimental studies as well as our FBA results (see Table 4) show that the two species are capable of metabolizing these byproducts in the presence of sufficient oxygen. Therefore, we investigated the impact of putative cross feeding by allowing each species to enhance its growth through uptake of the three byproducts (C-f scenario). The two species were assumed to uptake each byproduct with the same kinetics (see Table 2) due to lack of data on species and substrate specific uptake parameters.
The cross-feed scenario model predicted that W max remained 80 μm when byproduct cross feeding was incorporated. Because glucose was the more energetically favorable carbon source, most of the available oxygen was used for glucose oxidation and little oxygen remained for lactate oxidation. Contrary to our hypothesis, cross-feeding reduced the region where P. aeruginosa was present and did not substantially increase the P. aeruginosa biomass concentration within this region (Fig. 4a). To succinctly quantify this behavior, the P. aeruginosa and S. aureus biomass concentrations were averaged across the biofilm and compared to average concentrations obtained for the base case scenario without cross feeding. While the total biomass concentration was not affected, cross-feeding increased the fraction of S. aureus relative to P. aeruginosa (Fig. 4e). Since lactate was the primary byproduct of the two species biofilm, we attributed this behavior to S. aureus having more efficient lactate metabolism. Single species FBA results show that S. aureus has higher growth rates on lactate under oxygen sufficient and oxygen limited conditions (see Table 4). Because similar behavior was observed for glucose metabolism in single species biofilms, the addition of lactate consumption was predicted to further increased S. aureus dominance in the aerobic region where sufficient oxygen was available for lactate oxidation.
P. aeruginosa is known to secrete a number of small molecules including pyocyanin which inhibit and lyse competing bacteria such as S. aureus. Additional simulations were performed to explore the impact of a putative pyocyanin-mediated lysis mechanism on the two species biofilm. When this mechanism was combined with nutrient competition (Ly scenario), the model predicted that W max was slightly increased to 90 μm. Reduction of S. aureus biomass in the anaerobic region resulted in slightly higher glucose levels throughout the biofilm, allowing increased P. aeruginosa growth in the upper aerobic region and a greater biofilm thickness.
To allow direct comparison with the other species interaction scenarios, simulations also were performed for an 80 μm thick biofilm. Spatial profiles showed sharp partitioning of the two species with P. aeruginosa dominant in the upper aerobic region of the biofilm (Fig. 5a). This effect was achieved at the expense of the S. aureus biomass concentration, which was substantially reduced in the lower anaerobic region and dropped to zero at 40 μm. While the oxygen profile was largely unaffected, reduction of S. aureus biomass in the anaerobic region resulted in slightly higher glucose levels throughout the biofilm. Pyocyanin levels were highest in the aerobic region due to diffusion and removal at the tissue-biofilm interface. The metabolic burden of synthesizing pyocyanin was predicted to have a minimal effect on P. aeruginosa growth due to the small enforced bound of 0.1 mmol/gDW/h. At a maximum glucose uptake rate of 10 mmol/gDW/h, only 2.2 % of available carbon was used for pyocyanin synthesis.
As before, local effective growth rates and nutrient uptake rates were calculated at t = 10 h to investigate how pyocyanin affected multispecies metabolism as a function of position in the biofilm. In this case, the effective local growth rate of S. aureus was calculated by subtracting the maintenance energy associated death rate and the pyocyanin associated death rate from the biomass restricted growth rate at a given point z (see Eq. (2)). In the upper aerobic region, pyocyanin release caused the S. aureus death rate to exceed the growth rate (Fig. 5b) such that S. aureus was eventually eliminated from this region. This balance was reversed in the anaerobic region, although S. aureus growth was substantially reduced. By contrast, the P. aeruginosa growth rate was very similar to the pyocyanin-free case (see Fig. 3c). The long-term effects of these local growth rates were a sharp spatial partitioning of the two species and reduced S. aureus biomass concentrations in the anaerobic region. As a result, slightly higher glucose uptake rates were predicted across the biofilm while the oxygen uptake rates were similar to the pyocyanin-free case (see Fig. 3c). When biomass concentrations were averaged across the biofilm, the pyocyanin-mediated lysis mechanism was predicted to substantially increase P. aeruginosa competitiveness at the expense of both S. aureus and total biomass (Fig. 4e).
Combining byproduct cross feeding and the lysis mechanism (Ly + C-f scenario) did not change W max from the pyocyanin-free case (C-f scenario) but did increase S. aureus competitiveness by shifting the location where the species partitioned approximately 15 μm towards the biofilm-air interface (Fig. 4b). The addition of cross feeding resulted in average biomass concentrations that were roughly equal, while total biomass was reduced (Fig. 4e). We hypothesized that this unexpected effect was due to increased oxygen utilization by S. aureus for lactate oxidation. Although S. aureus biomass was simultaneously reduced by pyocyanin- mediated lysis, the oxygen used for S. aureus growth was not available for P. aeruginosa oxidative growth and total biomass decreased. Therefore, the pyocyanin mechanism was interpreted as an antagonistic mechanism by which P. aeruginosa increased its own competitiveness.
P. aeruginosa has a single flagellum that may allow motility in complex, heterogeneous environments such as biofilms  while S. aureus is generally viewed as non-motile . More specifically, P. aeruginosa has been observed to chemotax towards higher oxygen environments, a process known as aerotaxis, which offer more favorable growth conditions . To explore the impact of this putative aerotaxis mechanism on two species biofilm metabolism, the P. aeruginosa biomass equation included a chemotaxis term (see Eq. 2) and simulations were performed with both nutrient competition and aerotaxis (AT scenario). The energy requirements for chemotaxis were assumed negligible compared to growth. When aerotaxis was combined with nutrient competition, the model predicted that W max was increased to 120 μm. Aerotaxis increased spatial partitioning of the two species (see below) such that P. aeruginosa had access to more oxygen for lactate respiration, resulting in a thicker biofilm.
Spatial profiles generated for an 80 μm biofilm show almost complete partitioning of the two species, with P. aeruginosa dominating in the upper aerobic half, only S. aureus present in the lower anaerobic region, and the two species coexisting for about 10 μm near the middle of the biofilm (Fig. 6). Aerotaxis allowed P. aeruginosa to substantially improve its competitiveness by increasing its concentration in the upper portion of the biofilm rather than by moving the transition region between the two species (see Fig. 3a). When averaged across the biofilm, the biomass concentrations of the two species were approximately equal while total biomass was unaffected compared to the aerotaxis-free case (Fig. 4e). Unlike pyocyanin-mediated lysis, aerotaxis could be viewed as an antagonistic mechanism by which P. aeruginosa increased its own competitiveness without reducing total cell densities.
When byproduct cross feeding was added to P. aeruginosa aerotaxis and nutrient competition (AT + C-f scenario), the model predicted W max = 120 μm, the same value obtained in the absence of cross feeding. When simulations were performed for an 80 μm thick biofilm, the addition of cross feeding substantially increased S. aureus biomass in the biofilm while having a small negative impact on total biomass (Fig. 4e). Of the eight scenarios investigated, a maximum W max = 130 μm was predicted when nutrient competition and aerotaxis were combined with pyocyanin-mediated lysis of S. aureus (AT + Ly scenario). In this case, P. aeruginosa had access to increased glucose due to S. aureus death in the anaerobic region and increased lactate due to the absence of S. aureus in the aerobic region, which combined to enhance P. aeruginosa growth and allow a thicker biofilm. For a nominal biofilm thickness of 80 μm, only P. aeruginosa was present in the upper 50 μm of the biofilm and the amount of S. aureus was relatively small in the lower anaerobic region (Fig. 4d). The further addition of byproduct cross feeding did not affect W max but did increase S. aureus competitiveness (AT + Ly + C-f scenario) (Fig. 4e). Collectively, these predictions suggest that both pyocyanin-mediated lysis and aerotaxis are potentially powerful mechanisms for P. aeruginosa to enhance its competitiveness in multispecies biofilms with the faster growing facultative anaerobe S. aureus.
We hypothesized that the tendency of bacteria to self-organize into complex biofilm structures is at least partially driven by the appearance of metabolic niches . To test this hypothesis, we developed spatiotemporal metabolic models of chronic wound biofilms consisting of the aerobe Pseudomonas aeruginosa and/or the facultative anaerobe Staphylococcus aureus. The models were comprised of genome-scale metabolic reconstructions combined with partial differential equations (PDEs) for the diffusion and consumption/synthesis of nutrients/byproducts within the biofilm. Both single species and two species biofilms were simulated by supplying glucose at the tissue-biofilm interface (bottom of the biofilm) and oxygen at the biofilm-air interface (top of the biofilm).
Simulations of single species biofilms were performed to compare the metabolic behavior of each species and to provide a basis for comparing the two species biofilm simulations. Although the biofilm thickness was specified a priori for each simulation, we gained insights into the collective biofilm substrate utilization efficiency as quantified by the maximum achievable thickness W max under the specified environmental conditions. By assuming that the thickness was too large if the biomass concentration dropped below 10 g/L (5 % of the maximum value of 200 g/L) anywhere in the mature biofilm, the models predicted that S. aureus could grow much thicker biofilms with W max = 90 μm compared to P. aeruginosa with W max = 30 μm. Other differences predicted by the single species models were that S. aureus allowed much deeper oxygen penetration and produced much larger amounts of its primary metabolic byproduct lactate, a putative cross-fed metabolite. We attributed these differences in biofilm phenotypes to S. aureus having more efficient oxygen utilization in the upper aerobic region as well as substanially high growth rates in the lower anaerobic region. These model predictions could be experimentally tested by measuring metabolite concentration profiles using spatially resolved metabolomics [84–86] and gene expression profiles using spatially revolved transcriptomics [87, 88].
Two species biofilm simulations were performed for eight different scenarios to investigate the impact of putative mechanisms for interspecies interactions. For the base case involving only competition for the two nutrients glucose and oxygen, we found a maximum thickness W max = 80 μm for the two species biofilm. This value represented a linear combination of the single species W max values weighted by the average biomass concentrations in the two species biofilm. Dynamic simulations for an 80 μm thickness predicted that the two species would form a nearly mature biofilm in approximately 50 h, the time frame of many experimental studies [89, 90]. Two metabolic niches were quickly established in the simulated biofilm: an anoxic, glucose rich region in the lower half and an aerobic, glucose lean region in the upper half. Due to its superior anaerobic growth and more efficient oxygen utilization, S. aureus quickly established dominance throughout the biofilm. Steady-state spatial profiles demonstrated that this short-term behavior was maintained over long time periods, with P. aeruginosa only present in the aerobic region and lactate synthesized by S. aureus in the anaerobic region being the main byproduct. The experimental determination of spatially resolved biomass concentrations  would be beneficial in this context.
Cross feeding of secreted metabolic byproducts is common in bacterial communities [92, 93] and multispecies biofilms [94, 95]. For example, a cross feeding mechanism has been proposed for a polymicrobial infection system consisting of the two facultative anaerobes Aggregatibacter actinomycetemcomitans and Streptococcus gordonii . To explore the hypothesis that cross feeding would enhance P. aeruginosa competitiveness (C-f scenario), we allowed each species to uptake all possible carbon sources (glucose, lactate, succinate, acetate) with the same uptake kinetics. Therefore, the results reflected differences in carbon metabolism efficiency and not differences in uptake properties. The model predicted that cross feeding would not increase W max but would further enhance S. aureus competitiveness by reducing the region where P. aeruginosa was present. This prediction, which was consistent across all cross feeding scenarios investigated, was attributed to S. aureus having more efficient oxidative metabolism for the primary byproduct lactate. Byproduct cross feeding in P. aeruginosa/S. aureus chronic wound biofilms has not been experimentally studied to our knowledge and represents a promising area of research.
Cross feeding predictions were obtained assuming identical parameters for lactate uptake kinetics and non-growth associated ATP maintenance for the two species. Experimental studies with P. aeruginosa biofilms in the cystic fibrosis lung show that lactate actually can be a preferred carbon source to glucose [96, 97], suggesting enhanced lactate uptake capabilities. This environmental dependence emphasizes the importance of conducting uptake experiments and studying cross feeding under chronic wound relevant conditions. Well controlled planktonic growth experiments are needed to accurately estimate ATP maintenance demands of the two species, since lactate oxidation might confer a growth advantage to P. aeruginosa if the energetics are more favorable than those modeled.
P. aeruginosa secretes a wide variety of inhibitory compounds that have been shown to enhance its competitiveness against competing bacteria in multispecies biofilm communities [98–100]. To quantify the impact of a representative small molecule inhibitor, P. aeruginosa was forced to secrete pyocyanin and S. aureus was lysed by pyocyanin diffusing from high concentration regions. When combined with nutrient competition, the pyocyanin-mediated lysis mechanism produced a slightly larger W max = 90 μm and substantially increased P. aeruginosa competitiveness by inducing S. aureus death in the aerobic region. While we tuned our model to obtain reasonable extracellular pyocyanin concentrations , key parameters (i.e. synthesis rate, diffusion coefficient, killing rate) associated with the mechanism are unknown and need to be experimentally determined to accurately quantify the effect in chronic wound environments.
The motility of P. aeruginosa allows coordinated movement of cells towards higher oxygen levels through a chemotactic response known as aerotaxis . When putative P. aeruginosa aerotaxis was combined with nutrient competition (AT scenario), our biofilm model predicted substantially thicker biofilms (W max = 120 μm) and nearly complete species partitioning as observed experimentally [47, 101], with P. aeruginosa dominant in the upper aerobic half and only S. aureus present in the lower anaerobic region. While not producing as dramatic increase in the P. aeruginosa/S. aureus ratio as the pyocyanin mechanism, aerotaxis did not negatively impact total biomass within the biofilm. Therefore, we interpreted aerotaxis as a less antagonistic mechanism than pyocyanin secretion by which P. aeruginosa could enhance its own competitiveness. However, P. aeruginosa aerotaxis within biofilm environments has not been demonstrated to our knowledge. Therefore, biofilm reactor experiments aimed at demonstrating and quantifying the aerotactic response would be highly beneficial. Experimental testing could be achieved through a combination of traditional and spatially resolved omics technologies [86, 87, 102].
The two species system was predicted to support a maximum biofilm thickness much greater than P. aeruginosa alone but slightly less than S. aureus alone, suggesting an antagonistic metabolic effect of P. aeruginosa on S. aureus.
Nutrient gradients imposed by supplying glucose at the bottom and oxygen at the top of the biofilm induced spatial partitioning of the two species, with S. aureus most concentrated in the lower anaerobic region and P. aeruginosa present only in the upper aerobic region.
When each species was allowed to enhance its growth through consumption of secreted metabolic byproducts assuming identical uptake kinetics, the competitiveness of S. aureus was further enhanced due to its more efficient lactate oxidative metabolism.
Lysis of S. aureus by the small molecule inhibitor pyocyanin secreted from P. aeruginosa and/or P. aeruginosa aerotaxis towards high oxygen levels were predicted to enhance spatial portioning of the two species and to increase P. aeruginosa competitiveness in the aerobic region.
These model predictions require further validation through the execution of targeted experiments that augment existing results in the literature that support our conclusions. We believe that P. aeruginosa lysis of S. aureus combined with nutrient competition is a particularly relevant scenario for which model predictions could be tested experimentally.
The authors wish to thank Prof. Jason Papin (U. Virginia) for providing the P. aeruginosa metabolic reconstruction, Prof. Matthias Heinemann (U. Groningen) for providing the S. aureus metabolic reconstruction, and Prof. Paul Barton, Dr. Kai Höffner and Jose Gomez (MIT) for providing assistance with the DFAlab code.
DFBAlab is freely available for both education and nonprofit research purposes from https://yoric.mit.edu/dfbalab. The MATLAB simulation codes for the two species biofilm model can be downloaded from www.ecs.umass.edu/che/henson_group/downloads.html.
PP, RPC and MAH conceived the study. PP, JC and MAH developed the model and the model solution method. PP and MAH performed the simulations and analyzed the results. PP, JC, RPC and MAH prepared the manuscript. All authors read and approved the final manuscript.
Filkins LM, Graber JA, Olson DG, Dolben EL, Lynd LR, Bhuju S, O'Toole GA. Co-culture of Staphylococcus aureus with Pseudomonas aeruginosa drives S. aureus towards fermentative metabolism and reduced viability in a cystic fibrosis model. J Bacteriol. 2015. doi: 10.1128/JB.00059-15.


text:
seeds of Artemisia sieversiana, Sagebrush
Artemisia sieversiana, also known as the sagebrush is a plant species of the family Asteraceae. The plant's synonyms include Artemisia sieversiana Artemisia sieversiana var. sieversiana and Artemisia sieversiana f. sieversiana. Another common name for the plant is sierversian wormwood. Artemisia sieversiana has its origins in China. Artemisia sieversiana is an erect many-branched plant that can grow to a height of between 30 and 100 cm. The plant's leaves are hairy, ovate and alternate in the arrangement. Each leaf has a length of 1.5 to 15 cm and a width of 2 to 9 cm. Flowers are semi-spherical and have a diameter of 0.6 to 1.2 mm. Seeds are flat and oval. The plant has both male and female flowers. Insects are responsible for most of the pollination. The plant's leaves are a delicacy among many communities who regard them as ideal vegetables. An additional use of the plant is as a source of herbal remedies that have been in use for centuries. The remedies supposedly have antirheumatic, antithelmic, antiseptic, tonic and deobstruent attributes. Such qualities aid the remedies in not only curing the disease but also in preserving health. A common belief holds that the remedy can cure boils and relieve joint pains. Artemisia sieversiana is a hardy plant that is capable of growing under extreme temperature conditions and in poor soils. The plant gets more aromatic the more the soils around it get poorer. The plant is not only safe from poor soils and high temperatures, but also from most herbivorous animals that may want to consume it. The plant's strong aroma together with its bitter-tasting leaves wards off animals. Artemisia sieversiana thrives in well-drained soils that are either of neutral pH or slightly basic. The plant can easily cope with hot conditions and, therefore, prefers direct sunlight to growing in the shade.
You're reviewing:seeds of Artemisia sieversiana, Sagebrush
Gold Nugget Pumpkin seeds
Ipomoea nil Scarlett O'hara seeds
Cyphomandra betacea tree tomato "Tamarillo" (Solanum betacea) red
Humulus japonicus, Japanese hop
Aloe polyphylla - Spiral Aloe seeds, RARE
Citrus junos seeds
seeds of Pinus yunnanensis, Yunnan Pine
Strophanthus hispidius seeds
Cedrus deodora, Himalayan cedar seeds
Bubblegum 7 Pot, Chili pepper seeds
Salvia apiana, White sage, Sacred sage
spores of Cyathea dealbata, Silver Tree Fern

redpajama_set_name: RedPajamaCommonCrawl


Moose Juice is the best binding lubricant that was created by a competitive water skier for an ENVIRONMENTALLY SAFE way to ease into tight bindings. Moose Juice is fish freindly and contains no odor. It is designed to slip your foot in quick and wash out fast. The original GO FOR IT formula. Moose Juice is phosphate-free, odorless, bio-degradable, and slime-free. It will not damage your skin or harm our waterways. MooseJuice will create easy exit and entry into your water-sport bindings while optimizing performance. Contains NO moose!
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© 2013 Northshore Industries, Inc.. All Rights Reserved.


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Topic: James Bailey
James Bailey News
Odd News // 1 year ago
Doorbell camera records cow's late night front porch visit
A British Columbia couple whose doorbell camera issued an unusual number of overnight alerts checked the footage and identified the late night visitor to their front porch: an escaped cow.
Top News // 2 years ago
The Almanac
On July 4, 1939, Lou Gehrig gave his "luckiest man on the face of the Earth" speech in announcing his retirement from the New York Yankees.
In 1997, NASA's Pathfinder reached Mars to become the first U.S. spacecraft to land on the planet in more than two decades.
On July 4, 1910, African American boxer Jack Johnson took on former undefeated heavyweight champion James J. Jeffries, beating him in 15 rounds, to stake his claim as the as the greatest heavyweight in the world.
Did chase end Seattle-area crime spree?
A high-speed pursuit Tuesday night may have brought an end to a Seattle-area crime spree allegedly carried out by four suspects in the past month.
We're proud to support the intelligence community with products and services
Armedia software to support defense agency Mar 18, 2009
James M. Bailey is an American author and a TV actor. James Bailey is listed as James Michael Bailey on his acting credits ().
Bailey was born in Arkansas City, Kansas and completed high school in Westlake, Louisiana. Some time after this he went to Hollywood seeking an acting career where he went to acting school and eventually got some minor roles in television and films.
Not long after his arrival in Hollywood, Obsessive Compulsive Disorder (OCD) began to manifest along with severe depression and anxiety. Bailey suffered for many years before finally seeking psychiatric treatment for his emotional problems. He spent almost three months in an American clinic that specialized in OCD before his complex interactions with other patients led to him to be discharged.
It uses material from the Wikipedia article "James Bailey."


Casablanca (ADV) – The Presidency on Tuesday said there is no foreign pressure on President Muhammadu Buhari not to contest the 2019 presidential election.
Special Adviser to the President on Media and Publicity, Femi Adesina, in a statement described the story as fiction being paraded as news claiming that foreign powers, including the European Union, United States of America, United Kingdom, and Saudi Arabia, were putting pressure on Buhari not to seek re-election, due to some concocted reasons.
"If there are quarters in which President Buhari is held in very high esteem, and the work he is doing for Nigeria highly lauded, it is in the countries mentioned. Some of them are equally bastions of democracy, and will not interfere in the democratic affairs of a sovereign country. The newspaper should simply tell its story to the marines," Adesina said.


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Jupiter Goes Home to Sagittarius.
On November 8 Jupiter returned home to the constellation it feels most at home, Sagittarius. It takes Jupiter twelve years to go around the zodiac so it remains in a constellation just about a year.
For those with strong Jupiter/Sagittarius influence in their charts – Sun, Moon, ascendant – it is a time for growth, learning and reaching higher levels. Jupiter is constantly pushing us to go beyond boundaries – to see what is on the other side of fence. Set your goals for what you wish to overcome!
However, Jupiter is sometimes too big, too ambitious – and can lead to reckless behavior, drinking, gambling and other forms of risk taking. Jupiter-people sometimes think of themselves as too grand, too smart, too invincible. Keep your head on your shoulders!
For those with Sagittarius North or South Nodes this period is especially profound. If your North Node is involved this is a perfect time to work at living into it's positive energies. For South Node Sagittarius people, look at the downside – what can you leave behind? How can you make the leap into Gemini North Node while leaving behind the largess, aggrandizement and prevarications that come from a being too deep into your Sagittarius South.
Note: Donald Trump is a Sagittarius South Node. How he will react to this transit? Do you think he will take the route of leaving behind his negatives? Do you, really? Let's watch and see.


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The income tax brackets are slightly more for 2016 due to inflation, though tax rates don't change.
The 2016 standard deductions go up for heads of household $9,300 plus $1,550 more once they reach age 65. Married couples can take $12,600 plus $1,250 for each spouse age 65 or older. Single persons get $6,300 or $7,850 if they are 65 or older in 2016.


text:
Home » The President's Radio Address
The President's Radio Address
Weekly Compilation of Presidential Documents;12/24/2001, Vol. 37 Issue 51, p1799
Presents the text of a speech given by United States President George W. Bush, on December 15, 2001, which deals with the country's economic policy.
Remarks Following a Meeting With the Economic Team. Bush, George W. // Weekly Compilation of Presidential Documents;12/17/2002, Vol. 37 Issue 50, p1780
Presents the text of a speech given by United States President George W. Bush dated December 12, 2001 which deals with the economic stimulus package.
The President's Radio Address. Bush, George W. // Weekly Compilation of Presidential Documents;01/21/2002, Vol. 38 Issue 3, p59
Presents the text of a speech given by U.S. President George W. Bush on January 18, 2002 regarding the issues on economic policies and public expenditures.
A New International Economic Policy. Munk, Bernard E. // Orbis;Summer2001, Vol. 45 Issue 3, p401
Suggests agenda for the international economic policy (IEP) of United States (U.S.) President George W. Bush. Reason for the importance of a global macroeconomy; Coordinated policies for President Bush's agenda; Problem for the Bush administration's IEP; Proposed goal for President Bush's...
Is President W. repeating the sins of his father? // New Hampshire Business Review;04/20/2001-05/03/2001, Vol. 23 Issue 6, p13
Comments on the economic policy of United States President George W. Bush. Reason for proposing a tax cut; Lack of care about the loss of value in the Nasdaq composite index; Silence on the issue of low savings rate.
Remarks Prior to a Meeting With Senate Minority Leader Trent Lott and an Exchange With Reporters. Bush, George W. // Weekly Compilation of Presidential Documents;9/10/2001, Vol. 37 Issue 36, p1264
Presents the text of a speech given by United States President George W. Bush on September 4, 2001, which deals with the country's economic policy.
Remarks Prior to a Meeting With Congressional Leaders. Bush, George W. // Weekly Compilation of Presidential Documents;12/24/2001, Vol. 37 Issue 51, p1806
Presents the text of a speech given by United States President George W. Bush on December 19, 2001, which deals with the country's economic policy.
Statement on House of Representatives Action on the Economic Stimulus Package. Bush, George W. // Weekly Compilation of Presidential Documents;12/24/2001, Vol. 37 Issue 51, p1811
Presents the statement given by United States (U.S.) President George W. Bush on December 20, 2001, which deals with the U.S. House action on the 2002 economic stimulus package.
Bush tax cut could backfire in 2004. Cantor, Martin // Long Island Business News (7/1993 to 5/2009);5/9/2003, Vol. 50 Issue 20, p46A
Comments on the economic plan of U.S. President George W. Bush. Domination of the plan by a tax cut that Bush says would create jobs; Adverse effects of a decline in consumer spending; Federal spending cuts designed to match tax cuts; Analysis of the Bush plan by the Tax Policy Center at the...
Bush Is a Leader The Economy Can Trust. Mankiw, N. Gregory // Fortune;11/13/2000, Vol. 142 Issue 11, p60
Presents the author's perception of the benefits of United States presidential candidate George W. Bush's economic proposals. Details of Bush's tax cut plan; Efforts of Bush to move toward the privatization of Social Security; His stance on antitrust policy and school choice.


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Secret Visit (Egyptian Arabic: زيارة سرية translit: Zeyara Serreya) is a 1981 Egyptian film starring Salah Zulfikar, Athar El-Hakim, and Mahmoud El Meliguy. The film is written by Atef El-Ghamry, and directed by Nagy Anglo.

Synopsis
An accused who is sentenced to death by the judge Ismail, and all the evidence against him, but the accused denies committing the crime, which prompted his daughter to visit the judge in his home, and she is confident in the innocence of her father. The judge goes to the accused in a secret visit.

Cast

 Salah Zulfikar as Judge Ismail
 Athar El-Hakim as Nawal Attia Al-Nabrawi
 Mahmoud El-Meliguy as Attia Al-Nabawy
 Mariam Fakhr Eddine as Olfat, Ismail's wife
 Nabil Noureddine as Nabil Ismail
 Galal Issa as Ibrahim
 Samah Anwar as Noha Ismail
 Nabil El-Desouky as Owaidah El-Shamy
 Ali El-Sharif as coffee shop owner
 Abdul Rahim Al-Zarqani as Fahmy Al-Sayed
 Nadia Rafeeq as Nawal's mother
 Soad Hussein as Azhaar, Fahmy El-Sayed's wife

See also
 Salah Zulfikar filmography
 List of Egyptian films of 1981
 List of Egyptian films of the 1980s

References

External links

 Secret Visit on elCinema
 Shafik, Viola. "Youssef Chahine: DEVOURING MIMICRIES OR JUGGLING WITH SELF AND OTHER (EGYPT)." Ten Arab Filmmakers: Political Dissent and Social Critique, edited by Josef Gugler, Indiana University Press, 2015, pp. 98–121. JSTOR, www.jstor.org/stable/j.ctt16gzbbw.10. Accessed 6 Sept. 2021.

1981 films
1980s Arabic-language films
20th-century Egyptian films
Egyptian drama films
Films shot in Egypt

redpajama_set_name: RedPajamaWikipedia


text:
Q: JAVA After adding a seccond button and its actionListener, actions get executed twice After adding a listener to a second button, the first created button executes twice the same action:
public class ControladorTablaMaterial implements ActionListener {

 private VistaTablaMaterial vistaTablaMaterial;
    private JPanel jContentPane = null;
    private JScrollPane scrollPane = null;
    private JTable tablaMaterial;
    private JButton mostrarElementoButton;
    private JButton eliminarElementoButton;
    private ModeloTablaMaterial modeloTablaMaterial;

    public ControladorTablaMaterial(ArrayList<Material> coleccionMaterial, ActionListener listener) {
        String[] cabecera = {"Material", "Titulo"};
        this.vistaTablaMaterial = new VistaTablaMaterial(cabecera, coleccionMaterial);
        setupVistaTablaMAterial(listener);
    }

    private void setupVistaTablaMAterial(ActionListener listener) {
        this.scrollPane = vistaTablaMaterial.getScrollPane();
        this.tablaMaterial = vistaTablaMaterial.getTablaMaterial();
        this.modeloTablaMaterial = vistaTablaMaterial.getModeloTablaMaterial();
        this.mostrarElementoButton = vistaTablaMaterial.getMostrarElementoButton();
        this.eliminarElementoButton = vistaTablaMaterial.getMostrarElementoButton();
        this.initListeners(listener);
    }

    private void initListeners (ActionListener listener) {
        getMostrarElementoButton().addActionListener(listener);
        getEliminarElementoButton().addActionListener(listener);

    }

     @Override
    public void actionPerformed(ActionEvent e) {

    }

}

Everything works fine if I delete the line:
getEliminarElementoButton().addAtionListener(listener);

but of course I need that button to be listened to too.
Inside the listener class, in the actionPerformed(actionEvent e) method, I use the following code to differentiate both buttons:
if (e.getSource().equals(this.getControladorTablaMaterial().getMostrarElementoButton())) {

That seems to work fine except for this frame. Any guess?
Off topic: why isn't the code indentation working properly on Stackoverflow's editor?

A: The problem is in these lines:
this.mostrarElementoButton = vistaTablaMaterial.getMostrarElementoButton();
this.eliminarElementoButton = vistaTablaMaterial.getMostrarElementoButton();

That you are getting the same button for both.

redpajama_set_name: RedPajamaStackExchange


text:
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Witness & Grace Conversations
Democracy is collapsing, but let's imagine how the church could change that
Demonstrators marching together, while a person holds a sign that reads "We are on fire, act now." (Unsplash/Markus Spiske)
by Julia Walsh
Follow on Twitter at @juliafspa?lang=en
I was in another Zoom meeting with my friend, another Catholic sister who is a professor on a college campus. Together we were lamenting the extreme polarities in the church and society. Rubbing our foreheads. Sighing. Praying right out loud: God, help us.
Together we were bemoaning how the tensions impact our freedom to love and serve, to remain fierce in our hope. I asked her what she's seeing from her vantage as a professor who spends a lot of time with young adults. I know she is often buoyed by the zeal and vision of Generation Z. I was hoping to hear something encouraging.
But her expression turned grim as she started to speak about the conversations she's having with her colleagues — who are sociologists and historians.
"I hate to say it, but we're headed toward a civil war," she said.
She explained that because President Donald Trump's base will vote in person and Biden voters will vote by mail, we won't know the results quickly. Her colleagues are predicting that Trump will declare victory prematurely, and then his base will refuse to accept any other outcome.
The war might not be a bloody war but a culture war, she said — a power struggle fought across every sphere of society: online, in families, in churches and in the marketplace. Yet an image haunted me: those who say they have stockpiled guns emerging from the shadows, armed and angry, feeling entitled and emboldened to fight for what they believe belongs to them. Many will retaliate. Others will run and hide in fear. I can't imagine this scenario without bloodshed.
I realized, then, that this isn't too far from what we've already been going through. In the past decade, there's been an increase of militias organized in the backroads of the nation. Gun violence continues to rise, impacting both rural and urban dwellers. Since George Floyd's murder in May, a mixture of protest, counter-protest, looting and a militarized police presence have ripped wounds into our cities and stirred up doubts that downtowns can recover. In many places the battles continue. Meanwhile, we have surpassed 200,000 dead from COVID-19 this week, a harrowing juncture. This is the highest number among developed nations by far. It's embarrassing and a cause for reckoning, repenting and conversion. We are already fighting, and the body bags are piling up.
I'm afraid the war has already started. Its battles have already begun, with armed conflict and civil unrest exploding throughout the nation. Maybe the war is gradual and awful; much like the slow removal of a bandage from an infected wound. Some want monuments to come down; others are willing to use weapons to protect them. Some want to defund the police; others are still denying the reality of systemic racism in our police departments. Some wear masks to protect others from COVID19; others refuse to sacrifice their personal comfort.
We've sorted ourselves into echo-chambers online and in real life. We've succumbed to a mentality that our disagreements must make enemies. Moderates have become an endangered species, and middle grounds are the stuff for historical novels and nature preserves.
The good news is that we have a formula to move forward. We have inherited two frameworks that can help to restore civility and promote the common good, and liberty and justice for all.
First, the Constitution. Its authors were well aware of the fallen nature of humanity and offered us a path to uphold peace. The editors of America wrote recently:
The Constitution itself is inspired by a theological worldview. Our founders believed that they were creating a form of government for a fallen world. Their true genius lay in how they accounted for the human predilection toward sin and division by creating a strict separation of powers that, paradoxically, would serve to unify the country and guard it against would-be tyrants and demagogues.
Second, the Gospel of Jesus Christ and its accompanying Scriptures: from Matthew to Revelation, we are presented with a promise of covenantal love that reconciles differences and establishes peace and justice. In this vision, the church — the people of God — love their enemies, forgive wrongs, offer kindness and generosity, practice nonviolence, build relationships and act with charity and justice. The church needs to imagine how we can protect against the collapse of democracy and heed the call to be peacemakers. We must also heed the words of Scripture that remind us to persevere and be encouraging to one another. We may already be at war, and so we cannot delay in striving for reconciliation and peace.
History shows us that Christians have done this before. I'd like to offer one example that could be a map to start planning our route to reconciliation. During apartheid in South Africa, the South African Council of Churches work was twofold: they advocated for the end of apartheid and they began to prepare for the Truth and Reconciliation Commission.
Just two years after the election of Nelson Mandela, the commission began its proceedings. During its seven year duration, it received over 21,000 statements from victims and more than 7,000 amnesty applications; it granted 849 amnesties for crimes. Through each act of truth-telling, narratives were rewritten and civility was recovered. Forgiveness, transparency, lament and reconciliation helped move a nation closer to healing. Gradually the nation recovered.
When it comes to working to restore trust and unity, the church in America is uniquely positioned to end the collapse of democracy and bridge the divides that are destroying us. We are called to get to work, God's work, the work of love and mercy. Now is the time.
[Sr. Julia Walsh, Franciscan Sister of Perpetual Adoration, is a retreat director, speaker, educator, activist and award-winning writer who blogs at MessyJesusBusiness.com. Follow her on Twitter: @juliafspa.]
We all are called to community
USCCB official: The church must admit its role in destroying Native American culture
Basketball sisters head out on different paths, wind up on same journey 40 years later
LCWR leaders talk about unprecedented transformation of religious life
What part do Mary and the rosary play in a sister's vocation?
ByA Nun's Life Ministry
Black joy and 'The Embrace'
ByColleen Gibson
A new year invites us to see in new ways
ByNancy Sylvester
ByLinda Reeves, OSV News
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text:
Review: The Last Exorcism is a True Exercise in Great Horror
by Jeremy Kirk
Amidst the bland cavalcade of stagnant horror movies, there is one sub-genre (or film style, depending on how you view it) that doesn't seem to ever be tiring. No matter how stale the narratives get, the style of the mockumentary or "found footage" horror film hasn't hit the cinematic brick wall yet. Sure, there are those who complain about it, that it's overused or that it's never as effective as straight, smooth filmmaking. Yet, since The Blair Witch Project hit theaters in 1999, there have been films of this ilk that have consistently found their audience and brought out the best in fright the style has to offer.
The latest such film that rides right along that path is The Last Exorcism, a faux documentary style horror film that offers genuine scares, highly engrossing atmosphere, and a story that is as far from cookie cutter as you can get.
It's about much more than a documentary crew wanting to cover a priest's latest exorcism. The priest at the center of the film is Cotton Marcus, played by Patrick Fabian. He has performed over a hundred exorcisms on people who feel their bodies are being inhabited by demons or even the devil himself. The one thing about Cotton, though, is that he doesn't believe in demonic possession. He is a priest who, after the birth and eventual poor health of his son, is finding it harder and harder to believe in the higher power he so vehemently preaches. When it is learned that people may, in fact, be getting harmed from the fake exorcisms being performed, he decides it is time to shelve the holy water, hang up the cross, and allow a film crew to document just how he does it.
This is all built up in the film's first quarter, and it sets an incredible tone for the characters that are to be portrayed in the film. Nothing horrific occurs in the opening, and, if you didn't know precisely what film you were watching, you would swear you were watching a real documentary about a man who decides it is time to move on from the life he has been living.
Director Daniel Stamm and the screenwriters Huck Botko and Andrew Gurland do a fine job building Cotton's character up. Likewise, Fabian is completely solid in his portrayal here. Cotton is a character you sympathize with. You understand why he does what he does and why he feels he must come clean to the world about it. Even if you don't agree with his motives, you still have no qualms understanding them.
The real horrors come later as Cotton and the film crew travel to the titular last exorcism. It involves a young girl living with her father and brother deep in the back woods of Louisiana. They are a particularly devout family, so they have are fully in Cotton's hold as he goes through the motions of performing the exorcism on the girl. Then, after everything seems to go as planned, the real horrors begin.
The Last Exorcism is a horror film that utilizes both the commonplace jump scares but also one that builds a terrific atmosphere. The setting is muggy. The homes are the broken down homes of Louisiana you might see sitting in the tall grass as you travel the back roads of the deep South. It all feels real, so, once the scares begin, whether you know full well it is all made up or not, you are left unnerved and unsure of what is about to happen next.
Stamm and crew continuously move the story forward. It never gets bogged down in a swamp of needless exposition or revisiting of the same scare over and over again. Very early into the last half of the film, we realize full well the dangers involved in staying in the house and who the culprit is behind those dangers. In one of the film's more terrifying moments, the young girl, played to unsettling effect by Ashley Bell, takes the crew's camera with her on one of her late night escapades. What transpires is entirely horrifying yet cleverly shot in what it shows, what it doesn't show, what it keeps in full focus, and what it lets stay blurred out just enough. It's without a doubt the scariest moment of a film filled with heart-pounding terrors.
With an excellent setup, solid build throughout, and some of the best uses of atmosphere and sound seen in recent horror films, you would hope for a breathtaking yet absolutely revelatory ending. Unfortunately, that's not found here. It's on paper. What transpires really isn't the issue. It's the way it is presented, almost matter of factly that provides very little impact. When Blair Witch and Paranormal Activity had their final moments, they were shocking yet logical endings that left you completely uncomfortable, perfect for the drive home late at night. That same feeling is missing in The Last Exorcism, and, sadly, the best moments of the film are buried deep within the confines of the middle section.
It's also hard with these films to understand the camera man's motives. Sure, they're a film crew. They are filming a documentary. However, once someone begins chasing you with a shotgun or an ax, it's probably not the best time to worry about your Best Documentary Oscar acceptance speech, put down the damn camera, and run. Not found here, but, for the sake of capturing everything on film, it's almost forgivable.
The Last Exorcism is a breath of fresh air for horror fans, a highly effective thriller that offers great scares, great atmosphere, and presents a story that is much deeper than your average slasher or ghost story. It's a thrilling good time that makes you turn on the lights in your house once you get home from the theater, an effect that hasn't been found since… well, the last found footage movie about demons and exorcisms. It's a style of film making that is still in full swing, but, as long as the scares and story continue to hit as accurately as The Last Exorcism, it's not one I want exorcised out of the film industry for a long while.
Jeremy's Rating: 8 out of 10
Find more posts: Opinions, Review
I liked It!!
Adrian C. on Aug 31, 2010
i liked it to, the ending where they all die and u are left to wonder if the fire demon kills the preacher is very intriguing. I would have been really mad if it would have ended when they left the farm the last time. besides that cat getting massacred the rest was semi boring, but the ending was like -click- to me, it fulfilled the desires of a horror movie for me. HUGE fire demon, Satanic cults, the girls drawing of the preacher standing in front of the massive fire, the camera lady getting chopped into pieces, the camerman getting his head cut off, Pro ;D
Brutal on May 30, 2011
Super film! Eli Roth is a genius!
Lincoln on Aug 31, 2010
This was a crappy movie.
sharts on Aug 31, 2010
Ugh, really? You had to use a pun in the last line? And why is it you insist that the documentary style used in The Last Exorcism and Paranormal Activity is the only way to make a successful horror movie nowadays? If anything that particular style is, as you suggested, overused. Plus, it's manipulative. The only reason people go to see movies which employ such a style is for shock value. To put it another way, the only reason people go to see movies like that is the same reason people watch bum fights or be-headings on YouTube. I'm still going to go see this movie and appreciate it for what it is, but let's not claim that the success and artistry of a horror film is dependent on the documentary technique.
Bee on Aug 31, 2010
Well done, Mr. Kirk. You actually made me want to see this whereas before I had absolutely not interest. And I'm thinking that since my expectations for the end are so low I might actually enjoy it more.
Traveler on Aug 31, 2010
Good Movie but the Ending SUCKED ASS!
ModernRock on Aug 31, 2010
This was simply a bad movie. You've all seen the trailers, they make it out to be a horror movie along the lines of paranormal activity. What you don't realize, however, is that 90% of the material in the trailers is cut from a 10 minute segment of the movie and the rest of the film is entirely bereft of any fright factor whatsoever. I was actually more freaked out by the trailer for "Let Me In" they showed before the movie began. Furthermore, the ending was terrible. The movie raises quite a few questions that you'll expect an answer to, and then ends without answering a single one. It's as if the writer/director got bored after filming the first 90 minutes (talk about a short movie) and simply ended the movie in lieu of actually thinking up a fitting conclusion.
The G on Sep 1, 2010
@The G I haven't seen this yet but what you said reminds me of another major disappointment where the best scenes in the whole movie were in the trailer. Legions. God that movie sucked so bad. If you saw the trailer with the Granny in the diner and the ice cream truck man, you basically saw the best part of that crappy film.
JimD on Sep 1, 2010
Nowhere near as effective as Blair Witch Project or Paranormal Activity but def not the worst thing I've seen.
Alex on Sep 1, 2010
I liked this movie a lot the first time I saw it 20 years ago when it was called Rosemary's Baby.
Brian Ricci on Sep 1, 2010
@ The G I completely agree saw this last week and it was a complete let down. Ending was terrible, questions left unanswered and all the "scary" parts were shown in the trailer.
David on Sep 4, 2010
This I would have to disagree with you on, I was highly upset and disappointed with this crappy viral generation version of Rosemary's Baby. By the end of the movie I was really hoping that it would pay out and be worth the time I had already wasted. So I stay into the movie until near the end when they go back to the house after finally departing, which they were so desperate to do a few scenes prior, where they walk back into her doodled up room with a bunch of gibberish caked on the walls in which they couldnt even get the sigil of baphomet correct. At that point I was giggling until they wander out into the woods, where the Cotton character turns into Gandalf and walks towards the flames as if about to shout "YOU SHALL NOT PASS!," then I was just laughing my way to the exit. I just didnt buy it and it had lost all credibility at that point.
Hairu on Sep 6, 2010
Believing in God means believing in the existence of the devil. Spanish exorcist Fr. Jose Antonio Fortea Cucurull stated in an interview recently that though Satan does indeed exist, he does not have "horns, wings or a tail." Read this milestone book on demonology and diabolic possession by Fr. Gabriele Amorth, Rome's chief exorcist - An Exorcist Tells His Story Fr. Gabriele Amorth is also the honorary life president of the International Association of Exorcists, which he founded. Watch this ABC News' 20/20 documentary showing a real exorcism.
Andy Stone on Sep 30, 2010
Well, all things considered...
free rapidshare premium accounts on Dec 1, 2010
Ok, i don't know what the hell you have been smoking, Mr. Jeremy, but fuck you, this movie was a complete waste of fucking time. The trailer showed all the good parts, which were pretty lame, and the whole movie was like watching the shitfest called Quarantine only with more shit up the ass! Seriously, anyone who had to watch that movie because of reviews like this, i feel sorry for you for watching a movie that horrible, but anybody who liked it can kiss my ass.
Jacky on Feb 20, 2011
Interesting review, couple good points. I really enjoyed the ending and thought that it helped bring the movie together, even though it did raise a few questions (like how did the tape get from the camera to the media?), but it was definitely an entertaining horror flick. I gave the movie a B in my review for horror-movie.org, and I stand by the rating. You can read my review, if you're interested, at http://horror-movie.org/wp/2011/05/the-last-exorcism/
horror-movie.org on May 8, 2011


In November a mission followed in Strean Presbyterian Church, but the attendance was so great, the meetings were moved to the larger First Presbyterian Church. One elderly man who remembered the 1859 revival, said that some of the effects of the Holy Spirit's working even exceeded what happened in '59.
A band of seventy voluntary workers from the various churches were employed as ushers, enquiry room attendants, booksellers and pew watchers. It was the duty of the latter to get in touch with any person who appeared to be anxious and seek to lead them out. I would say in this respect I had the great joy of seeing two to whom I spoke turn back and wait for the after-meeting, at which they came out for the Lord. One of these is a very bright young Christian and a really active worker. For about three weeks there was an average of fifty each night passing through the enquiry room, some truly remarkable cases being amongst these. That the Lord is no respecter of persons was clearly seen by the varied types of sinners who sought Eternal Life. On the last Sunday, exactly 117 signed the decision card. The latter is similar to that used in 1859. After the mission, 350 decision card tear-offs were sent to the Presbyterian Church, and 250, 140 and 100 to three other churches in the town.
From 'All for Jesus' by Stanley Barnes, published by Ambassador Productions, p64.


text:
After President Trump's announcement of his intent to renegotiate the North American Free Trade Agreement (NAFTA) in May 2017, CDM launched a trinational, coordinated campaign with allied organizations from Mexico, the United States, and Canada to strengthen protections for migrant workers in the renegotiation process.
CDM has supported migrant workers in filing multiple petitions under the NAFTA labor side accord. Through this experience, we have seen firsthand NAFTA's failure to protect and vindicate the rights of migrant workers who rightfully claim that the US government has failed to enforce its labor laws. Worker petitioners under the existing agreement have waited years for responses to their complaints, and the remedies offered by the U.S. government have not made workers or their communities whole. Through supporting workers like Adareli Ponce Hernandez and Leonardo Cortez Vitela, CDM knows that NAFTA's complaint and dispute resolution mechanisms are deeply flawed.
The work ahead is clear: we will keep the pressure on the Trump administration to protect migrant workers in NAFTA renegotiation. Through reports, supplements to our previous petitions, and educational advocacy, we will continue to demand that the renegotiated NAFTA protect migrant workers.
Recent Actions
On March 23, 2021, migrant worker women submitted the first-ever petition against the U.S. under the USMCA. The petition was signed by a binational coalition of dozens of allies led by CDM. Together, we're arguing that the United States government failed to live up to its obligations under the trade agreement by allowing systemic gender-based discrimination against migrant worker women.
On June 30, almost four years after submitting the complaint and hours before the new treaty came into effect, the Mexico National Administrative Office finally issued a report on our complaint, stating that they "recognize the existence of mechanisms in the United States, to which people can go to manifest the facts that they consider discriminatory." The Office absurdly offered that workers can call ICE to report abusive employers or turn to brochures and posters about how to submit complaints to federal government agencies. This response is frustrating and disappointing.
On August 28, 2018, we celebrated a big win: announced that the preliminary agreement for a new trade deal reached by the United States and Mexico will ensure migrant workers are protected under labor laws, bring labor obligations from a side accord into the core of the agreement, and make labor obligations fully enforceable.
On August 17, 2018, we delivered a letter to the USTR on behalf of 30 organizations recommending that labor protections, including those for the tens of thousands of migrant workers, be strong, enforceable and binding in the new NAFTA. On August 21, we delivered it to the Ministry of Economy in Mexico City.
On January 22, 2018, on the eve of the sixth round of the renegotiation, a group of migrant worker women and supporting nongovernmental organizations across the trinational trade corridor took the next step in holding the United States government accountable for violating its obligations under the NAFTA labor side accord. The group argues that the United States fails to protect women and remedy labor rights violations in temporary labor migration programs.
On December 13, 2017, we presented our report, Coerced under NAFTA: Abuses of Migrant Workers in the TN Visa Program and Recommendations for Reform. This report highlights the range of abuses to which thousands of Mexican professionals are subjected each year under the TN visa program, created under NAFTA, and recommends actions the governments should take to address them. Workers experience fraud, economic coercion, and retaliation, the root of which is employer control of the visa and hiring processes.
On October 17, 2017, in line with the fourth round of NAFTA negotiations, CDM submitted a petition signed by 98 individuals and organizations, urging the USTR to protect migrant workers in the renegotiation.
On June 28, 2017, CDM, the International Labor Recruitment Working Group, and petitioners Adareli Ponce Hernandez and Leonardo Cortez Vitela testified before an interagency panel at the U.S. International Trade Commission. We demanded that a renegotiated NAFTA must include all current NAFTA labor principles, as well as additional protections for internationally recruited workers; eliminate barriers to accessing justice and benefits; and include a clear and enforceable labor dispute resolution process.
On June 27, 2017, CDM met with key members of the Senate Finance and House Ways and Means Committees, which have jurisdiction over the negotiations.
On June 26, 2017 — the eve of the U.S. Trade Representative's hearing on NAFTA negotiating objectives –, CDM organized allied workers, organizations, and academics from the U.S., Canada, and Mexico for a day-long strategy meeting in Washington, D.C. to create a coordinated strategy for demanding migrant worker protections in all three countries.
On June 12, 2017, CDM submitted comments in response to the U.S. Trade Representative's request for input on the NAFTA negotiating objectives. We recommended the inclusion of a NAFTA labor chapter that would use the same dispute settlement mechanism as other complaints, rather than a labor side accord, along with other recommendations that would strengthen the complaint and dispute mechanisms and bolster legal protections for migrant workers.


Your Opinions On Cross Platform Play?..
Feel sorry for the PS4 and PC players tbh.
E60 wrote Feel sorry for the PS4 and PC players tbh.
It's neat, they've been adding it to a few games lately, I have it enabled on Gears of War 4, Rocket League, and mine craft as of recently.


text:
Erik Thomas Cole (* 6. November 1978 in Oswego, New York) ist ein ehemaliger US-amerikanischer Eishockeyspieler, der im Verlauf seiner aktiven Karriere zwischen 1996 und 2015 unter anderem 938 Spiele für die Carolina Hurricanes, Edmonton Oilers, Canadiens de Montréal, Dallas Stars und Detroit Red Wings in der National Hockey League auf der Position des linken Flügelstürmers bestritten hat. Mit den Carolina Hurricanes gewann Cole im Jahr 2006 den Stanley Cup.

Karriere
Cole begann seine Laufbahn im Jahr 1996 in der United States Hockey League als Juniorenspieler bei den Des Moines Buccaneers. Von dort wechselte er ins nordamerikanische Collegeeishockey an die Clarkson University. In dieser Zeit wurde er von den Carolina Hurricanes beim NHL Entry Draft 1998 in der zweiten Runde an 71. Stelle gezogen. Hier schaffte Cole es mehrmals in eine All-Star-Auswahl. Als vollwertiger Profi spielte er dann in der International Hockey League IHL bei den Cincinnati Cyclones, den Partnern der Carolina Hurricanes. In seiner zweiten Spielzeit konnte er 23 Tore schießen und spielte sich damit zur Saison 2001/02 zu den Hurricanes in die NHL.

Seine erste NHL-Saison konnte Cole als fünftbester Torschütze unter den NHL-Neulingen beenden. Außerdem konnte er in dieser Saison mit den Hurricanes die Finalserie der Play-offs um den Stanley Cup erreichen, die man allerdings gegen die Detroit Red Wings verlor. In der kommenden Saison konnte Cole aufgrund von Verletzungen nicht in allen Spielen antreten. In der Spielzeit 2003/04 erreichte er dann 42 Punkte.

Den Lockout der NHL-Saison 2004/05 verbrachte Cole in Deutschland bei den Eisbären Berlin in der Deutschen Eishockey Liga, wo er den Meistertitel feiern konnte. In den Play-offs konnte Cole fünf Tore und einen Assist erzielen. Im dritten Spiel der Best-of-Five-Serie in der letzten Play-off-Runde der Eisbären gegen die Adler Mannheim konnte Cole einen Unterzahltreffer erzielen und lieferte so den Ausgleich, den die Eisbären dann zum Sieg ausbauten, zu dem Cole noch einen weiteren Treffer beitrug. Negative Aufmerksamkeit erregte Cole, als er im Play-off-Viertelfinale gegen die Augsburger Panther den Verteidiger Arvīds Reķis hart gegen den Kopf checkte, so dass Rekis auf dem Eis aufschlug und sich eine ernste Gehirnerschütterung zuzog. Cole sollte für sechs Spiele gesperrt werden und zur Zahlung einer Strafe 1800 Euro verpflichtet werden. Nach dem Einspruch der Eisbären wurde Cole noch für vier Spiele gesperrt. Zwar entschuldigte sich Cole in aller Öffentlichkeit bei Rekis, erklärte aber ebenso, dass er nie die Absicht hatte, seinen Gegenspieler zu verletzen. Die Fans der Eisbären Berlin selbst zeigten öffentlich Kritik, indem sie in Absprache mit dem Club während der Sperre bei der Mannschaftsaufstellung der Eisbären immer den Namen "Cole" riefen. In der folgenden Saison kehrte Cole zu den Carolina Hurricanes zurück. Er hatte bis zuletzt immer wieder betont, dass wenn seine Leistungen einmal nicht mehr für die NHL reichen sollten, er eine Rückkehr nach Berlin nicht ausschließen würde.

In der regulären Saison konnte er acht Mal das spielentscheidende Tor schießen. Im März 2006 erlitt Cole bei einem Check von hinten gegen die Bande durch Brooks Orpik, Verteidiger der Pittsburgh Penguins, einen Bruch eines Halswirbels, zuvor hatte Cole im gleichen Spiel zum zweiten Mal hintereinander zwei Tore in einem Spiel erzielen können. Orpik wurde für drei Spiele gesperrt. Erst im sechsten Spiel der letzten Play-off-Serie 2006 konnte Cole auf das Eis zurückkehren und gewann mit den Hurricanes den Stanley Cup. Trotz des verletzungsbedingten Ausfalls konnte er mit 30 Toren und 29 Assists auf seine bisher beste NHL Saison zurückblicken. Im Sommer 2006 erhielt er von den Hurricanes einen Vertrag über weitere drei Jahre.

Am 1. Juli 2011 unterzeichnete Cole einen Kontrakt für vier Jahre bei den Montréal Canadiens. Im Februar 2013 wurde er zu den Dallas Stars transferiert, im Austausch wechselte Michael Ryder – gemeinsam mit einem Drittrunden-Wahlrecht im NHL Entry Draft 2013 – zu den Montréal Canadiens.

Nach knapp zwei Jahren wurde Cole gemeinsam mit einem leistungsabhängigen Drittrunden-Wahlrecht im NHL Entry Draft 2015 zu den Detroit Red Wings transferiert. Im Gegenzug erhielt Dallas Mattias Bäckman, Mattias Janmark und ein Zweitrunden-Wahlrecht im Draft 2015. Bis zum Saisonende kam Cole auf elf Einsätze für die Red Wings, die danach bekanntgaben, seinen Vertrag nicht zu verlängern. Im September 2017 gab Cole schließlich seinen Rücktritt vom aktiven Sport zurück.

International
Cole vertrat die USA bei der Weltmeisterschaft 2005. Hier erzielte er in sieben Spielen ein Tor und fünf Assists und erreichte mit seinem Team den sechsten Platz. Bei den Olympischen Winterspielen 2006 erzielte er ein Tor und ein Assist. Die USA erreichte dabei den achten Platz.

Erfolge und Auszeichnungen

Karrierestatistik

International
Vertrat die USA bei:
 Weltmeisterschaft 2005
 Olympischen Winterspielen 2006
 Weltmeisterschaft 2007

Weblinks

Eishockeynationalspieler (Vereinigte Staaten)
Eishockeyspieler (Clarkson Golden Knights)
Eishockeyspieler (Cincinnati Cyclones, IHL)
Eishockeyspieler (Carolina Hurricanes)
Eishockeyspieler (Eisbären Berlin)
Eishockeyspieler (Edmonton Oilers)
Eishockeyspieler (Canadiens de Montréal)
Eishockeyspieler (Dallas Stars)
Eishockeyspieler (Detroit Red Wings)
Olympiateilnehmer (Vereinigte Staaten)
Teilnehmer der Olympischen Winterspiele 2006
Stanley-Cup-Sieger
Deutscher Meister (Eishockey)
US-Amerikaner
Geboren 1978
Mann


Our kids event will be a fun swim/run race. All the kids will get times and show up in results just like all the other athletes.
The run will be up to a turn point on the promenade then straight to the finish.