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Accurate and robust finite element methods for computing flows with differential constitutive equations require approximation methods that numerically preserve the ellipticity of the saddle point problem formed by the momentum and continuity equations and give numerically stable and accurate solutions to the hyperbolic constitutive equation. We present a new finite element formulation based on the synthesis of three ideas: the discrete adaptive splitting method for preserving the ellipticity of the momentum/continuity pair (the DAVSS formulation), independent interpolation of the components of the velocity gradient tensor (DAVSS-G), and application of the discontinuous Galerkin (DG) method for solving the constitutive equation. We call the method DAVSS-G/DG. The DAVSS-G/DG method is compared with several other methods for flow past a cylinder in a channel with the Oldroyd-B and Giesekus constitutive models. Results using the Streamline Upwind Petrov–Galerkin method (SUPG) show that introducing the adaptive splitting increases considerably the range of Deborah number (De) for convergence of the calculations over the well established EVSS-G formulation. When both formulations converge, the DAVSS-G and DEVSS-G methods give comparable results. Introducing the DG method for solution of the constitutive equation extends further the region of convergence without sacrificing accuracy. Calculations with the Oldroyd-B model are only limited by approximation of the almost singular gradients of the axial normal stress that develop near the rear stagnation point on the cylinder. These gradients are reduced in calculations with the Giesekus model. Calculations using the Giesekus model with the DAVSS-G/DG method can be continued to extremely large De and converge with mesh refinement.

Septic shock is a leading cause of mortality in intensive care units. No new interventions in the last 20 years have made a substantial impact on the outcome of patients with septic shock. Identification of inhibitable pathways that mediate death in shock is an important goal. Two novel caspase inhibitors, (2-indolyl)-carbonyl-Ala-Asp-fluoromethylketone (IDN 1529) and (1-methyl-3-methyl-2-indolyl)-carbonyl-Val-Asp-fluoromethylketone (IDN 1965), were studied in a murine model of endotoxic shock. IDN 1529 prolonged survival when given before or up to 3 hr after high-dose LPS (p < 0.01) and increased by 2.2-fold the number of animals surviving longterm after a lower dose of LPS (p < 0.01). Despite its similar chemical structure, IDN 1965 lacked these protective effects. Both compounds inhibited caspases 1, 2, 3, 6, 8, and 9, and both afforded comparable reduction in Fas- and LPS-induced caspase 3-like activity and apoptosis. Paradoxically, administration of IDN 1529 but not IDN 1965 led to an increase in the LPS-induced elevation of serum cytokines related directly (IL-1β, IL-18) or indirectly (IL-1α, IL-1Ra) to the action of caspase 1. A process that appears to be distinct from both apoptosis and the release of inflammatory cytokines is a late-acting requirement for lethality in endotoxic shock. Inhibition of this process can rescue mice even when therapy is initiated after LPS has made the mice severely ill.

Ultimately, numerical simulation of viscoelastic flows will prove most useful if the calculations can predict the details of steady-state processing conditions as well as the linear stability and non-linear dynamics of these states. We use finite element spatial discretization coupled with a semi-implicit θ -method for time integration to explore the linear and non-linear dynamics of two, two-dimensional viscoelastic flows: plane Couette flow and pressure-driven flow past a linear, periodic array of cylinders in a channel. For the upper convected Maxwell (UCM) fluid, the linear stability analysis for the plane Couette flow can be performed in closed form and the two most dangerous, although always stable, eigenvalues and eigenfunctions are known in closed form. The eigenfunctions are non-orthogonal in the usual inner product and hence, the linear dynamics are expected to exhibit non-normal (non-exponential) behavior at intermediate times. This is demonstrated by numerical integration and by the definition of a suitable growth function based on the eigenvalues and the eigenvectors. Transient growth of the disturbances at intermediate times is predicted by the analysis for the UCM fluid and is demonstrated in linear dynamical simulations for the Oldroyd-B model. Simulations for the fully non-linear equations show the amplification of this transient growth that is caused by non-linear coupling between the non-orthogonal eigenvectors. The finite element analysis of linear stability to two-dimensional disturbances is extended to the two-dimensional flow past a linear, periodic array of cylinders in a channel, where the steady-state motion itself is known only from numerical calculations. For a single cylinder or widely separated cylinders, the flow is stable for the range of Deborah number (De) accessible in the calculations. Moreover, the dependence of the most dangerous eigenvalue on De≡ λV / R resembles its behavior in simple shear flow, as does the spatial structure of the associated eigenfunction. However, for closely spaced cylinders, an instability is predicted with the critical Deborah number De c scaling linearly with the dimensionless separation distance L between the cylinders, that is, the critical Deborah number De L c ≡ λV / L is shown to be an O (1) constant. The unstable eigenfunction appears as a family of two-dimensional vortices close to the channel wall which travel downstream. This instability is possibly caused by the interaction between a shear mode which approaches neutral stability for De ≫ 1 and the periodic modulation caused by the presence of the cylinders. Nonlinear time-dependent simulations show that this secondary flow eventually evolves into a stable limit cycle, indicative of a supercritical Hopf bifurcation from the steady base state.

The mechanism by which membrane-bound Bcl-2 inhibits the activation of cytoplasmic procaspases is unknown. Here we characterize an intracellular, membrane-associated form of procaspase-3 whose activation is controlled by Bcl-2. Heavy membranes isolated from control cells contained a spontaneously activatable caspase-3 zymogen. In contrast, in Bcl-2 overexpressing cells, although the caspase-3 zymogen was still associated with heavy membranes, its spontaneous activation was blocked. However, Bcl-2 expression had little effect on the levels of cytoplasmic caspase activity in unstimulated cells. Furthermore, the membrane-associated caspase-3 differed from cytosolic caspase-3 in its responsiveness to activation by exogenous cytochrome c. Our results demonstrate that intracellular membranes can generate active caspase-3 by a Bcl-2–inhibitable mechanism, and that control of caspase activation in membranes is distinct from that observed in the cytoplasm. These data suggest that Bcl-2 may control cytoplasmic events in part by blocking the activation of membrane-associated procaspases.

Bcl-xL, an antiapoptotic member of the Bcl-2 family, inhibits programmed cell death in a broad variety of cell types. Recent reports have demonstrated that cytochrome c is released from mitochondria during apoptosis and have suggested that this release may be a critical step in the activation of proapoptotic caspases and subsequent cell death. Furthermore, it has been demonstrated that Bcl-2 can prevent the release of cytochrome c from mitochondria in cells triggered to undergo apoptosis. This has led to the hypothesis that the antiapoptotic effects of Bcl-2 family members are due specifically to their ability to prevent cytochrome c release thus preventing subsequent cytochrome c-dependent caspase activation. In the present report, we use microinjection techniques to investigate the relationship between cytochrome c release, induction of apoptosis, and Bcl-xL activity in intact cells. We demonstrate that microinjection of cytochrome c into the cytosol of human kidney 293 cells results in a dose-dependent induction of apoptosis. In contrast, MCF7 breast carcinoma cells (stably transfected to express the Fas antigen CD95, and denoted MCF7F) that lack detectable levels of caspase 3 (CPP32), are totally resistant to microinjection of cytochrome c. However, transfection of MCF7F cells with an expression plasmid coding for pro-caspase 3, but not other pro-caspases, restores cytochrome c sensitivity. Although MCF7F cells are insensitive to cytochrome c microinjection, they rapidly undergo apoptosis in a caspase-dependent manner in response to either tumor necrosis factor or anti-Fas plus cycloheximide, and these deaths are strongly inhibited by Bcl-xL expression. Furthermore, microinjection of cytochrome c does not overcome these antiapoptotic effects of Bcl-xL. Our results support the concept that the release of cytochrome c into the cytoplasm can promote the apoptotic process in cells expressing pro-caspase 3 but that cytochrome c release is not sufficient to induce death in all cells. Importantly, the ability of Bcl-xL to inhibit cell death in the cytochrome c-insensitive MCF7F cells cannot be due solely to inhibition of cytochrome c release from mitochondria.

Fas- and tumor necrosis factor receptor 1 (TNFR1)-induced apoptosis is mediated by the interaction of FADD with caspase-8. Here, we report that the bovine herpesvirus 4 (BHV4) BORFE2 gene encodes a protein that inhibits Fas- and TNFR1-induced apoptosis and contains death effector domains (DEDs). Using the yeast two-hybrid system, we found that the BORFE2 protein interacts with the prodomain of caspase-8. Furthermore, we show that BHV4 BORFE2 is a member of a family of DED-containing proteins that includes other gamma-2 herpesviruses, such as Kaposi's sarcoma-associated herpesvirus and herpesvirus saimiri.

Local similarity solutions are presented for the stress field of a fluid described by the Oldroyd-B viscoelastic constitutive equation near the singularity caused by the intersection of a planar free surface and a solid surface along which Navier’s slip law holds, the partial-slip/slip problem. For the case where the velocity field is given by Newtonian kinematics, the elastic stress field is predicted to have a logarithmic singularity as the point of attachment of the free surface is approached. Asymptotic analysis for the fully-coupled flow, where the stress and flow fields are determined simultaneously, results in a local form for the flow and elastic stress fields that is similar in form to that for the decoupled case. For both the coupled and decoupled flow problems, the strength of the singularity depends on the dimensionless solvent viscosity and the slip coefficient, but not upon the Deborah number. The asymptotic results for the coupled flow differ from the predictions with Newtonian kinematics i...

We identified and cloned a novel human protein that contains FADD/Mort1 death effector domain homology regions, designated FLAME-1. FLAME-1, although most similar in structure to Mch4 and Mch5, does not possess caspase activity but can interact specifically with FADD, Mch4, and Mch5. Interestingly, FLAME-1 is recruited to the Fas receptor complex and can abrogate Fas/TNFR-induced apoptosis upon expression in FasL/tumor necrosis factor-sensitive MCF-7 cells, possibly by acting as a dominant-negative inhibitor. These findings identify a novel endogenous control point that regulates Fas/TNFR1-mediated apoptosis.

The local solution behavior near corners formed by the intersection of a slip surface with either a no-slip or a shear-free boundary is analyzed by finite element calculations of the two-dimensional flow of an inertialess Newtonian fluid in several model flow geometries; these flows are the flow in a tapered contraction, a sudden expansion and the extrudate swell from a planar die. Local finite element mesh refinement based on irregular, embedded elements is used to obtain extremely fine resolution of the velocity and pressure fields near the region where there is a sudden change in boundary condition. The calculations accurately reproduce the expected asymptotic behavior for a shear-free surface intersecting a no-slip boundary, where the solution is given by a self-similar form for the velocity and pressure fields. Replacing the shear-free condition with a slip condition yields a similar form for the local velocity and pressure fields and indicates that the slip boundary behaves, to leading order, as a shear-free surface. Calculations for a slip boundary intersecting a shear-free surface yield similar results, with the local behavior being given by asymptotic analysis for two shear-free surfaces intersecting to form a wedge. These results suggest that replacing the no-slip boundary condition in planar Newtonian die swell with a slip boundary condition can give rise to local behavior of velocity gradients and pressure which is more singular than the flow created with no-slip boundary conditions. This prediction is confirmed by calculations of Newtonian die swell with slip. These calculations also demonstrate that the local solution in Newtonian die swell is sensitive to the details of the numerical method.

To identify novel antiapoptotic proteins encoded by DNA viruses, we searched viral genomes for proteins that might interfere with Fas and TNFR1 apoptotic signaling pathways. We report here that equine herpesvirus type 2 E8 protein and molluscum contagiosum virus MC159 protein both show sequence similarity to the death effector domains (DEDs) of the Fas/TNFR1 signaling components FADD and caspase-8. Yeast two-hybrid analysis revealed that E8 protein interacted with the caspase-8 prodomain whereas MC159 protein interacted with FADD. Furthermore, expression of either E8 protein or MC159 protein protected cells from Fas- and TNFR1-induced apoptosis indicating that certain herpesviruses and poxviruses use DED-mediated interactions to interfere with apoptotic signaling pathways. These findings identify a novel control point exploited by viruses to regulate Fas- and TNFR1-mediated apoptosis.

Neuronal apoptosis occurs during nervous system development and after pathological insults to the adult nervous system. Inhibition of CED3/ICE-related proteases has been shown to inhibit neuronal apoptosisin vitroandin vivo, indicating a role for these cysteine proteases in neuronal apoptosis. We have studied the activation of the CED3/ICE-related protease CPP32 in twoin vitromodels of mouse cerebellar granule neuronal cell death: K+/serum deprivation-induced apoptosis and glutamate-induced necrosis. Pretreatment of granule neurons with a selective, irreversible inhibitor of CED3/ICE family proteases, ZVAD-fluoromethylketone, specifically inhibited granule neuron apoptosis but not necrosis, indicating a selective role for CED3/ICE proteases in granule neuron apoptosis. Extracts prepared from apoptotic, but not necrotic, granule neurons contained a protease activity that cleaved the CPP32 substrate Ac-DEVD-aminomethylcoumarin. Induction of the protease activity was prevented by inhibitors of RNA or protein synthesis or by the CED3/ICE protease inhibitor. Affinity labeling of the protease activity with an irreversible CED3/ICE protease inhibitor, ZVK(biotin)D-fluoromethylketone, identified two putative protease subunits, p20 and p18, that were present in apoptotic but not necrotic granule neuron extracts. Western blotting with antibodies to the C terminus of the large subunit of mouse CPP32 (anti-CPP32) identified p20 and p18 as processed subunits of the CPP32 proenzyme. Anti-CPP32 specifically inhibited the DEVD-amc cleaving activity, verifying the presence of active CPP32 protease in the apoptotic granule neuron extracts. Western blotting demonstrated that the CPP32 proenzyme was expressed in granule neurons before induction of apoptosis. These results demonstrate that the CED3/ICE homolog CPP32 is processed and activated during cerebellar granule neuron apoptosis. CPP32 activation requires macromolecular synthesis and CED3/ICE protease activity. The lack of CPP32 activation during granule neuron necrosis suggests that proteolytic processing and activation of CED3/ICE proteases are specific biochemical markers of apoptosis.

Purpose FMS-like tyrosine kinase 3–internal tandem duplication (FLT3-ITD) mutations in acute myeloid leukemia (AML) are associated with early relapse and poor survival. Quizartinib potently and selectively inhibits FLT3 kinase activity in preclinical AML models. Patients and Methods Quizartinib was administered orally at escalating doses of 12 to 450 mg/day to 76 patients (median age, 60 years; range, 23 to 86 years; with a median of three prior therapies [range, 0 to 12 therapies]), enrolled irrespective of FLT3-ITD mutation status in a phase I, first-in-human study in relapsed or refractory AML. Results Responses occurred in 23 (30%) of 76 patients, including 10 (13%) complete remissions (CR) of any type (two CRs, three CRs with incomplete platelet recovery [CRp], five CRs with incomplete hematologic recovery [CRi]) and 13 (17%) with partial remissions (PRs). Of 17 FLT3-ITD–positive patients, nine responded (53%; one CR, one CRp, two CRis, five PRs); of 37 FLT3-ITD–negative patients, five responded (14%; two CRps, three PRs); of 22 with FLT3-ITD–indeterminate/not tested status, nine responded (41%; one CR, three CRis, five PRs). Median duration of response was 13.3 weeks; median survival was 14.0 weeks. The most common drug-related adverse events (> 10% incidence) were nausea (16%), prolonged QT interval (12%), vomiting (11%), and dysgeusia (11%); most were ≤ grade 2. The maximum-tolerated dose was 200 mg/day, and the dose-limiting toxicity was grade 3 QT prolongation. FLT3-ITD phosphorylation was completely inhibited in an in vitro plasma inhibitory assay. Conclusion Quizartinib has clinical activity in patients with relapsed/refractory AML, particularly those with FLT3-ITD, and is associated with an acceptable toxicity profile.

Recent evidence suggests that CPP32 is an essential component of an aspartate-specific cysteine protease (ASCP) cascade responsible for apoptosis execution in mammalian cells. Activation of CPP32 could lead to activation of other downstream ASCPs, resulting in late morphological changes such as lamin cleavage and DNA fragmentation, observed in cells undergoing apoptosis. Here we describe the identification and cloning of a novel human ASCP named Mch6 from Jurkat T lymphocytes. We demonstrate that the pro-enzymes of Mch6 and the lamin-cleaving enzyme Mch2alpha are substrates for mature CPP32. Site-directed mutagenesis revealed that CPP32 processes pro-Mch6 preferentially at Asp330 to generate two subunits of molecular masses 37 kDa (p37) and 10 kDa (p10). However, CPP32 processes pro-Mch2alpha at three aspartate processing sites (Asp23, Asp179, and Asp193) to produce the large (p18) and small (p11) subunits of the mature Mch2alpha enzyme. The CPP32-processed Mch2alpha is capable of cleaving the VEIDN lamin cleavage site, indicating that CPP32 can, in fact, activate pro-Mch2alpha. Granzyme B at a concentration that allows processing and activation of CPP32 failed to process pro-Mch2alpha. However, incubation of pro-Mch2alpha with granzyme B in the presence of a cellular extract containing pro-CPP32 resulted in activation of pro-CPP32 and subsequent processing of pro-Mch2alpha. Interestingly, granzyme B can also process pro-Mch6 but at a site N-terminal to that cleaved by CPP32. These data suggest that Mch2alpha and Mch6 are downstream proteases activated in CPP32- and granzyme B-mediated apoptosis. This is the first demonstration of a protease cascade involving granzyme B, CPP32, Mch2alpha, and Mch6 and evidence that the lamin-cleaving enzyme Mch2 is a target of mature CPP32.

Emerging evidence suggests that an amplifiable protease cascade consisting of multiple aspartate specific cysteine proteases (ASCPs) is responsible for the apoptotic changes observed in mammalian cells undergoing programmed cell death. Here we describe the cloning of two novel ASCPs from human Jurkat T-lymphocytes. Like other ASCPs, the new proteases, named Mch4 and Mch5, are derived from single chain proenzymes. However, their putative active sites contain a QACQG pentapeptide instead of the QACRG present in ail known ASCPs. Also, their N termini contain FADD-like death effector domains, suggesting possible interaction with FADD. Expression of Mch4 in Escherichia coli produced an active protease that, like other ASCPs, was potently inhibited (Kj = 14 nM) by the tetrapeptide aldehyde DEVD-CHO. Interestingly, both Mch4 and the serine protease granzyme B cleave recombinant proCPP32 and proMch3 at a conserved IXXD-S sequence to produce the large and small subunits of the active proteases. Granzyme B also cleaves proMch4 at a homologous IXXD-A processing sequence to produce mature Mch4. These observations suggest that CPP32 and Mch3 are targets of mature Mch4 protease in apoptotic cells. The presence of the FADD-like domains in Mch4 and Mch5 suggests a role for these proteases in the Fas-apoptotic pathway. In addition, these proteases could participate in the granzyme B apoptotic pathways.

This study involves an examination of the changes in the rheological behavior (specifically the non-Newtonian viscosity) of blends of various starches with a synthetic polymer (polyethylene-co-vinyl alcohol) as a result of irradiation-modification. The results are generally consistent with fact that starch degrades substantially upon irradiation, while the synthetic polymer is relatively unaffected. However, the high-amylose starch-containing blends are less sensitive to irradiation than we would expect ; this is attributed to possible interactions between the linear molecules from the high-amylose starch and the synthetic polymer.

Irradiation-modification of the blends of various starches with a synthetic polymer [poly(ethylene-co-vinyl alcohol)] was carried out using an electron beam. The effect of irradiation on neat starches was studies using gel permeation chromatography. Changes in the thermal and mechanical properties of the blends, as well as in their microstructures, were also evaluated. The data indicate, consistent with other reports in literature, that starch molecules fragment under the effect of ionizing radiation, while the EVOH is relatively unaffected. These substantial (mainly physical) modifications to the starch molecules manifest themselves in changes in the thermal behavior of the blends. Furthermore, the mechanical properties of filaments obtained from molten irradiated pellets were quite different from those of control filaments, at least for some starches. Micrographic examination of some blends indicated a correspondence between a modification in the microstructure of the filaments and a change in their mechanical properties. It seems likely that the enhanced mobility of the fragmented starch molecules in the melt is responsible for these changes in the microstructure and concomitantly, the mechanical properties of the blend. Such an irradiation-based physical modification of starch may be of use in tailoring the properties of commercial blends of starches with synthetic thermoplastics. © 1996 John Wiley & Sons, Inc.

The human proto-oncogene bcl-2 and its Caenorhabditis elegans homologue ced-9 inhibit programmed cell death. In contrast, members of the human interleukin-1beta converting enzyme (ICE) family of cysteine proteases and their C. elegans homologue CED-3 promote the death program. Genetic experiments in C. elegans have shown that ced-9 is formally a negative regulator of ced-3 function, but neither those studies nor others have determined whether CED-9 or Bcl-2 proteins act biochemically upstream or downstream of CED-3/ICE proteases. CPP32, like all known members of the CED-3/ICE family, is synthesized as a proenzyme that is subsequently processed into an active protease with specificity for cleavage at Asp-X peptide bonds. In this report, we demonstrate that the CPP32 proenzyme is proteolytically processed and activated in Jurkat cells induced to die by Fas ligation. CPP32 activation is blocked by cell-permeable inhibitors of aspartate-directed, cysteine proteases, suggesting that pro-CPP32 is cleaved by active CPP32 or by other ICE family members. Heterologous expression of Bcl-2 in Jurkat cells prevents Fas-induced cell death as well as proteolytic processing and activation of CPP32. Thus, Bcl-2 acts at or upstream of the CPP32 activation step to inhibit apoptosis induced by Fas stimulation.

This study tests the survey responses of clients of accounting firms to determine what marketing cues influence their selection of firms and what relationship these cues have on the perceived service quality of the firms' products. The study argues that three marketing cue factors are influential in the clients' selection of accounting firms. Using Principle Components Analysis, three factors were identified: Factor 1, termed Internal Factors-Tangibles includes the following cues: fees, physical facilities, appearance/dress of employees, convenience of location, and available parking. Factor 2, termed Internal Factors-Intangibles includes reputation of the firm, professional quality of employees, and “my own personal experience with other accounting firms.” Factor 3, termed External Factors comprises the opinion of friends/relatives, manner of employees, and word-of-mouth reputation. The relationship betwecn the resultant cues and perceived service quality measures suggests that the intangible an...

Laser Doppler velocimetry and flow‐induced birefringence are used to measure the rate of deformation and the principal components of the refractive index tensor in a 5% polyisobutylene (PIB) solution in tetradecane (C14) flowing along the centerplane of an abrupt 3.97:1 planar contraction. The stress optical law is used to interpret the birefringence data in terms of the normal stress difference, which is used to calculate a transient elongational viscosity defined along the centerplane. These measurements are compared directly to predictions of six multimode, differential constitutive models (Oldroyd‐B, White–Metzner, Acierno et al., Giesekus, Bird–DeAguiar, and Phan‐Thien–Tanner) that are fit to steady and small amplitude oscillatory shear flow data for the PIB/C14 solution. The fluid exhibits slight elongational thickening followed by apparent extensional thinning at higher elongation rates. We believe that this ‘‘thinning’’ behavior is due to the decreased residence time of the polymer molecules in the high‐strain‐rate region as the flow rate (and maximum elongation rate) is increased. The nonlinear constitutive equations, except for the White–Metzner model, are virtually indistinguishable in their description of the dynamical response of the fluid in this experiment; however, the Phan‐Thien–Tanner model gives the best quantitative fit to the data. These results point to the need for experiments in which the fluid flowing along the centerline is subjected to a greater total elongational strain.

The two‐dimensional, free‐surface flow of a Newtonian fluid exiting from a planar die is computed by finite element analysis using quasiorthogonal mesh generation and local mesh refinement with irregular, embedded elements to obtain extreme resolution of the velocity and pressure fields near the die edge, where the fluid sheet attaches to the solid boundary. Calculations for the limit of large surface tension, the stick‐slip problem, reproduce the singular behavior near the die edge expected from asymptotic analysis using a self‐similar form for the velocity field. Results for finite capillary number (Ca) predict that the meniscus separates from the die at a finite contact angle and suggest that the capillary force enters the dominant normal stress balance at the die edge through an infinite curvature, as previously suggested by Schultz and Gervasio. The size of this region with large positive curvature increases with increasing Ca, and the strength of the singularity is in good agreement with theoretical predictions for a straight meniscus attached to the die at the appropriate contact angle predicted by the simulations. The contact angle appears to be determined from matching of the inner solution structure valid near the singularity with the bulk flow, in agreement with arguments made by Ramalingam; increasing the Reynolds number decreases the contact angle, corroborating this effect. Introducing fluid slip along the surface of the die changes the structure of the singularity in the pressure and stresses, but does not alleviate the singular behavior. In fact, the calculations with slip coefficients small enough not to change the bulk solution are more difficult than calculations with the no‐slip boundary condition.

A new mixed finite element method is presented that has improved numerical stability and has similar numerical accuracy compared to the EVSS/FEM developed by Rajagopalan et al. (J. Non-Newtonian Fluid Mech. 36 (1990) 159–192). The new method, denoted EVSS-G/FEM, is based on separate bilinear interpolation of all relevant components of the velocity gradient tensor. Lagrangian bilinear approximations of this variable are formed by least-squares interpolation of the derivatives of a Lagrangian biquadratic approximation to the velocity field that is generated by solution of the momentum and continuity equations using a standard mixed formulation for velocity and pressure. Elastic components of the stress are computed by solving the constitutive equation using either the streamline upwinding (SU) or streamline upwind Petrov-Galerkin (SUPG) methods with bilinear interpolation. The motivation for the interpolation of the velocity gradients and for the stress interpolation is to force the compatibility between these variables that is needed to satisfy differential viscoelastic constitutive equations in the limit where the velocity field vanishes, as it does near solid boundaries. The enhanced numerical stability of the EVSS-G/FEM over the EVSS/FEM formulation is demonstrated for the calculation of the linear stability of planar Couette flow of a UCM fluid, where closed form results exist for the linear stability of the steady-state flow; hence, any instability detected in the numerical solution is a result of either the finite element approximation or the time integration method. The second possibility is eliminated by using a fully implicit time integration method. Calculations with the EVSS-G/FEM are stable for De >100, whereas calculations with the EVSS/FEM become numerically unstable for De >5. The EVSS-G formulation is combined with finite element discretizations that incorporate local adaptive mesh refinement for increasing the accuracy of the calculations in regions where the stress and velocity gradients change rapidly. The accuracy of the EVSS-G/FEM is demonstrated by steady-state calculations for the flow between eccentric rotating cylinders, flow through a wavy-walled tube, and for flow through a square array of cylinders; each has been used before as a test problem for the accuracy of viscoelastic flow calculations. As in previous calculations, discretization of the constitutive equation by SUPG gives superior accuracy at high values of De compared to the application of SU.

Blends consisting of a systematic compositional series of starch/poly(ethylene-co-vinyl alcohol) were studied by using capillary viscometry. Three different types of starches were used: waxy maize, a starch containing essentially 100% amylopectin; native corn, possessing approximately 70% amylopectin and 30% amylose; and a treated, high-amylose starch, consisting of approximately 30% amylopectin and 70% amylose. Poly(ethylene-co-vinyl alcohol) with 44 mol% ethylene was blended in varying proportions with the starches, and glycerine and water were added as plasticizers. The viscosity of the blends was measured over the shear-rate range 1–1000 s−1. Samples unconditioned (as stored) as well as conditioned at 65% relative humidity and 23°C were measured at temperatures between 140 and 170°C. All viscosity results fall in the power-law regime, and there is no indication of approach to a zero-shear-rate viscosity in any of the starch blend systems. As in starch-based food materials, the viscosity has an Arrhenius dependence on temperature, an exponential dependence on moisture content and a power-law dependence on shear rate. The power-law exponents from the starch blends are less than unity, indicating that the melts are shear thinning. Results on the starch/poly(ethylene-co-vinyl alcohol) blends indicate that the viscosity decreases with decreasing amylose content or increasing poly(ethylene-co-vinyl alcohol) content. The shear-rate dependence increases with an increase in the amylose content of the blends.

A series of potent, selective platelet-derived growth factor receptor-family kinase inhibitors was optimized starting from a globally selective lead molecule 4 through structural modifications aimed at improving the physiochemical and pharmacokinetic properties, as exemplified by 18b. Further clearance reduction via per-methylation of the α-carbons of a solubilizing piperidine nitrogen resulted in advanced leads 22a and 22b. Results from a mouse tumor xenograft, a collagen-induced arthritis model, and a 7 day rat in vivo tolerability study culminated in the selection of compound 22b (AC710) as a preclinical development candidate.

Formal methods describe a class of system analysis techniques that seek to prove specific propertiesabout analyzed designs, or locate flaws compromising those properties. As an analysis capability,these techniques are the subject of increased interest fromboth internal and external customersof Sandia National Laboratories. Given this lab's other areas of expertise, Sandia is uniquelypositioned to advance the state-of-the-art with respect toseveral research and application areaswithin formal methods. This research project was a one-yeareffort funded by Sandia's CyberSecurity S&T Investment Area in its Laboratory Directed Research&Development program toinvestigate the opportunities for formal methods to impactSandia's present mission areas, morefully understand the needs of the research community in the area of formal methods and whereSandia can contribute, and clarify from those potential research paths those that would best advancethe mission-area interests of Sandia. The accomplishmentsfrom this project reinforce the utilityof formal methods in Sandia, particularly in areas relevantto Cyber Security, and set the stagefor continued Sandia investments to ensure this capabilityis utilized and advanced within thislaboratory to serve the national interest.4

Mutations in the BRAF gene have been identified in approximately 7% of cancers, including 60% to 70% of melanomas, 29% to 83% of papillary thyroid carcinomas, 4% to 16% colorectal cancers, and a lesser extent in serous ovarian and non–small cell lung cancers. The V600E mutation is found in the vast majority of cases and is an activating mutation, conferring transforming and immortalization potential to cells. CEP-32496 is a potent BRAF inhibitor in an in vitro binding assay for mutated BRAFV600E (Kd BRAFV600E = 14 nmol/L) and in a mitogen-activated protein (MAP)/extracellular signal–regulated (ER) kinase (MEK) phosphorylation (pMEK) inhibition assay in human melanoma (A375) and colorectal cancer (Colo-205) cell lines (IC50 = 78 and 60 nmol/L). In vitro, CEP-32496 has multikinase binding activity at other cancer targets of interest; however, it exhibits selective cellular cytotoxicity for BRAFV600E versus wild-type cells. CEP-32496 is orally bioavailable in multiple preclinical species (>95% in rats, dogs, and monkeys) and has single oral dose pharmacodynamic inhibition (10–55 mg/kg) of both pMEK and pERK in BRAFV600E colon carcinoma xenografts in nude mice. Sustained tumor stasis and regressions are observed with oral administration (30–100 mg/kg twice daily) against BRAFV600E melanoma and colon carcinoma xenografts, with no adverse effects. Little or no epithelial hyperplasia was observed in rodents and primates with prolonged oral administration and sustained exposure. CEP-32496 benchmarks favorably with respect to other kinase inhibitors, including RAF-265 (phase I), sorafenib, (approved), and vemurafenib (PLX4032/RG7204, approved). CEP-32496 represents a novel and pharmacologically active BRAF inhibitor with a favorable side effect profile currently in clinical development. Mol Cancer Ther; 11(4); 930–41. ©2012 AACR.

Finite‐amplitude waves propagating at constant speed down an inclined fluid layer are computed by finite element analysis of the Navier–Stokes equations written in a reference frame translating at the wave speed. The velocity and pressure fields, free‐surface shape and wave speed are computed simultaneously as functions of the Reynolds number Re and the wave number μ. The finite element results are compared with predictions of long‐wave, asymptotic theories and boundary‐layer approximations for the form and nonlinear transitions of finite‐amplitude waves that evolve from the flat film state. Comparisons between the finite element calculations and the long‐wave predictions for fixed μ and increasing Re agree well for small‐amplitude waves. However, for larger‐amplitude waves the long‐wave results diverge qualitatively from the finite element predictions; the long‐wave theories predict limit points in the solution families that do not exist in the finite element solutions. Comparisons between the finite ele...

The Ras/RAF/MEK/ERK mitogen-activated protein kinase (MAPK) signaling pathway plays a central role in the regulation of cell growth, differentiation, and survival. Expression of mutant BRAF(V600E) results in constitutive activation of the MAPK pathway, which can lead to uncontrolled cellular growth. Herein, we describe an SAR optimization campaign around a series of quinazoline derived BRAF(V600E) inhibitors. In particular, the bioisosteric replacement of a metabolically sensitive tert-butyl group with fluorinated alkyl moieties is described. This effort led directly to the identification of a clinical candidate, compound 40 (CEP-32496). Compound 40 exhibits high potency against several BRAF(V600E)-dependent cell lines and selective cytotoxicity for tumor cell lines expressing mutant BRAF(V600E) versus those containing wild-type BRAF. Compound 40 also exhibits an excellent PK profile across multiple preclinical species. In addition, significant oral efficacy was observed in a 14-day BRAF(V600E)-dependent human Colo-205 tumor xenograft mouse model, upon dosing at 30 and 100 mg/kg BID.

We present a rigorous mathematical framework for analyzing dynamics of a broad class of Boolean network models. We use this framework to provide the first formal proof of many of the standard critical transition results in Boolean network analysis, and offer analogous characterizations for novel classes of random Boolean networks. We precisely connect the short-run dynamic behavior of a Boolean network to the average influence of the transfer functions. We show that some of the assumptions traditionally made in the more common mean-field analysis of Boolean networks do not hold in general. For example, we offer some evidence that imbalance, or expected internal inhomogeneity, of transfer functions is a crucial feature that tends to drive quiescent behavior far more strongly than previously observed., To appear as a Letter in Physical Review Letters 8 pages, 4 figures

Aryl phenyl ureas with a 4-quinazolinoxy substituent at the meta-position of the phenyl ring are potent inhibitors of mutant and wild type BRAF kinase. Compound 7 (1-(5-tert-butylisoxazol-3-yl)-3-(3-(6,7-dimethoxyquinazolin-4-yloxy)phenyl)urea hydrochloride) exhibits good pharmacokinetic properties in rat and mouse and is efficacious in a mouse tumor xenograft model following oral dosing.

Mixed finite-element methods for computation of viscoelastic flows governed by differential constitutive equations vary by the polynomial approximations used for the velocity, pressure, and stress fields, and by the weighted residual methods used to discretize the momentum, continuity, and constitutive equations. This paper focuses on computation of the linear stability of the planar Couette flow as a test of the numerical stability for solution of the upper-convected Maxwell model. Previous theoretical results prove this inertialess flow to be always stable, but that accurate calculation is difficult at high De because eigenvalues with fine spatial structure and high temporal frequency approach neutral stability. Computations with the much used biquadratic finite-element approximations for velocity and deviatoric stress and bilinear interpolation for pressure demonstrate numerical instability beyond a critical value of De for either the explicitly elliptic momentum equation (EEME) or elastic-viscous split-stress (EVSS) formulations, applying Galerkin's method for solution of the momentum and continuity equations, and using streamline upwind Petrov-Galerkin (SUPG) method for solution of the hyperbolic constitutive equation. The disturbance that causes the instability is concentrated near the stationary streamline of the base flow. The removal of this instability in a slightly modified form of the EEME formulation suggests that the instability results from coupling the approximations to the variables. A new mixed finite-element method, EVSS-G, is presented that includes smooth interpolation of the velocity gradients in the constitutive equation that is compatible with bilinear interpolation of the stress field. This formulation is tested with SUPG, streamline upwinding (SU), and Galerkin least squares (GLS) discretization of the constitutive equation. The EVSS-G/SUPG and EVSS-G/SU do not have the numerical instability described above; linear stability calculations for planar Couette flow are stable to values of De in excess of 50 and converge with mesh and time step. Calculations for the steady-state flow and its linear stability for a sphere falling in a tube demonstrate the appearance of linear instability to a time-periodic instability simultaneously with the apparent loss of existence of the steady-state solution. The instability appears as finely structured secondary cells that move from the front to the back of the sphere.

Laser Doppler velocimetry (LDV) and video flow visualization are used to investigate the creeping motion of a highly elastic, constant-viscosity fluid flowing past a cylinder mounted centrally in a rectangular channel. A sequence of viscoelastic flow transitions are documented as the volumetric flow rate past the cylinder is increased and elastic effects in the fluid become increasingly important. Velocity profiles clearly show that elasticity has almost no effect on the kinematics upstream of the cylinder, but that the streamlines in the wake of the cylinder are gradually shifted further downstream . Finite element calculations with a nonlinear constitutive model closely reproduce the evolution of the steady two-dimensional velocity field. However, at a well defined set of flow conditions the steady planar stagnation ow in the downstream wake is experimentally observed to become unstable to a steady, three-dimensional cellular structure. The Reynolds number at the onset of the flow instability is less than 0.05 and inertia plays little role in the flow transition, LDV measurements in the wake close to the cylinder reveal large spatially periodic fluctuations of the streamwise velocity that extend along the length of the cylinder and more than five cylinder radii downstream of the cylinder. Fourier analysis shows that the characteristic spatial wavelength of these flow perturbations scales closely with the cylinder radius R . Flow visualization combined with LDV measurements also indicates that the perturbations in the velocity field are confined to the narrow region of strongly extensional flow near the downstream stagnation point. A second flow transition is observed at higher flow rates that leads to steady translation of the cellular structure along the length of the cylinder and time-dependent velocity oscillations in the wake. Measurements of the flow instability are presented for a range of cylinder sizes, and a stability diagram is constructed which shows that the onset point of the wake instability depends on both the extensional deformation of the fluid in the stagnation flow and the shearing flow between the cylinder and the channel.

Finite element analysis is used to compute the inertialess flow of a viscoelastic fluid around a sphere falling in a cylindrical tube. Calculations are reported for three differential constitutive models with constant viscosities: the Upper-Convected Maxwell model (UCM), the Oldroyd-B model (OLDB) and the dumbbell model of Chilcott and Rallison (CR). Calculations are based on the Explicitly Elliptic Momentum Equation (EEME) and the Elastic Viscous Split Stress (EVSS) methods for calculations with models without and with a Newtonian solvent contribution, respectively. The calculations converged with mesh refinement for all three models for values of De below 1.6. Calculations with the UCM and OLDB models are limited below De = 2 by loss of convergence of the Newton iteration. Mesh refinement does not seem to alleviate these limits. Calculations with the CR model and moderate values of the maximum extensibility of the dumbbell L converge to higher values of De ; however, computations for large values of L require fine meshes in the stagnation region behind the sphere.

A decoupled finite-element method (INT/FEM) is presented for calculation of two-dimensional viscoelastic flows with integral constitutive models. The momentum and continuity equations are solved by Galerkin's method with the viscoelastic stress treated as a fixed body force. The viscoelastic stress is computed by using the stream function to track fluid particles upstream, integrating a system of ordinary differential equations that govern the displacement-gradient tensor, and evaluating the integral constitutive equation by numerical quadrature. The quasi-linear upper-convected Maxwell and Oldroyd-B models, as well as the nonlinear model of Papanastasiou, Scriven and Macosko (PSM), are used in the simulations. The efficiency of the integral method is compared to that of the recently developed finite-element method (EVSS/FEM) for differential constitutive models. Convergence and accuracy of the INT/FEM are shown by calculations for flow between eccentric cylinders. The upper limit in De attainable by using the INT/FEM is comparable to values for the EVSS/FEM only for constitutive models with a shear-thinning ratio of the first normal stress difference to the shear stress and a large solvent contribution to the solution viscosity. The INT/FEM becomes the more efficient technique for simulation with this type of constitutive equation when three or more relaxation modes are included in the memory function.

The goal of this research was to explore first principles associated with mixing of diverse implementations in a redundant fashion to increase the security and/or reliability of information systems. Inspired by basic results in computer science on the undecidable behavior of programs and by previous work on fault tolerance in hardware and software, we have investigated the problem and solution space for addressing potentially unknown and unknowable vulnerabilities via ensembles of implementations. We have obtained theoretical results on the degree of security and reliability benefits from particular diverse system designs, and mapped promising approaches for generating and measuring diversity. We have also empirically studied some vulnerabilities in common implementations of the Linux operating system and demonstrated the potential for diversity to mitigate these vulnerabilities. Our results provide foundational insights for further research on diversity and redundancy approaches for information systems.

Finite‐element predictions of steady, two‐dimensional viscoelastic flow based on the elastic‐viscous split stress formulation of Rajagopalan et al. [Rajagopalan, D., R. C. Armstrong, and R. A. Brown, ‘‘Finite Element Methods for Calculation of Steady, Viscoelastic Flow Using Constitutive Equations with a Newtonian Viscosity,’’ J. Non‐Newt. Fluid Mech. 36, 159–192 (1990)] are compared to birefringence measurements of stress for flow between eccentric rotating cylinders of a solution of polystyrene in tricresyl phosphate. Steady and small‐amplitude oscillatory shearing measurements are used to characterize the shear flow rheology of the fluid, and a four‐mode Giesekus model with a Newtonian solvent viscosity is fit to the rheological data. Comparison of the stress fields measured by flow‐induced birefringence in the one‐dimensional flow between concentric cylinders to the mechanically measured viscometric data show that the optical data compare reasonably well with the mechanical data. Comparison of optical...

The purpose of this work is to extend a theoretical model of layered (laminar) media for SHS combustion presented in an earlier article [1] to explore possible mechanisms for after-burning in SHS (i.e., gasless) combustion. As before, our particular interest is how the microscopic geometry of the solid reactants is reflected in the combustion wave and in the reaction product. The model is constructed from alternating lamina of two pure reactants that interdiffuse exothermically to form a product. Here, the laminar model is extended to contain layers of differing thicknesses. Using asymptotic theory, it was found that under certain conditions, the combustion wave can become “detached,” and an initial thin flame propagates through the media, leaving a slower, thicker flame following behind (i.e., afterburning). Thin laminae are consumed in the initial flame and are thick in the secondary. The thin flame has a width determined by the inverse of the activation energy of diffusion, as found previously. The width of the afterburning zone, however, is determined by the absolute time of diffusion for the thicker laminae. Naturally, when the laminae are all the same thickness, there is only one thin flame. The condition for the appearance of afterburning is found to be contingent on the square of the ratio of smallestto-largest thicknesses being considerably less than unity.

The EEME/finite-element method is used to compute steady flows of fluids with constitutive behavior described by the Modified Upper Convected Maxwell Model of Apelian and a modified form of the dumbbell model of Chilcott and Rallison through abrupt, axisymmetric contractions. Both constitutive models predict constant viscosity and shear thinning first normal stress coefficient, but differ qualitatively in the behavior of the elongational viscosity. Asymptotic analysis for both models predicts that the solution has Newtonian-like spatial structure near the reentrant corner and integrable stresses and velocity gradients there. With the Newtonian-like asymptotics, the stress field can be approximated by conventional Lagrangian finite elements and computed by the streamline upwind Petrov Galerkin (SUPG) method. The finite element calculations are stable and convergent: higher values of Deborah number are reached with increasing mesh refinement. Moreover, the predicted asymptotic structure of the stress and velocity fields is recovered near the corner in the calculations. Calculations with both constitutive equations show the stretching of the Newtonian corner vortex toward the reentrant corner and its growth upstream with increasing Deborah number for 4:1 and 8:1 contraction ratios. The characteristics of the computed vortex are in semi-quantitative agreement with experiments for Boger fluids for which the flow is axisymmetric and steady.

Time-dependent simulations using the EEME finite-element method are reported for calculation of the linear stability and nonlinear dynamics of the transition to time-periodic, viscoelastic flow in axisymmetric TaylorCouette flow between parallel cylinders. The linear stability analysis is based on time integration of the linearized finite-element equations and reproduces the oscillatory linear instability recently analyzed by Larson et al. past a critical Deborah number Dec. Linear analysis presented here shows that the viscoelastic instability corresponds to a secondary flow structure composed of multiple toroidal flow cells nested radially, and that these cells travel across the gap between the cylinders. Nonlinear simulations demonstrate the existence of supercritical, i.e. for De - Dec > 0, time-periodic flow states. For only small changes in De >Dec, the flatness of the neutral stability curve with variation in the flow cell height leads to nonlinear interactions between flows that are closely spaced in De. These interactions will make the observation of simple oscillations near onset extremely difficult.

A constitutive equation for computing particle concentration and velocity fields in concentrated monomodal suspensions is proposed that consists of two parts: a Newtonian constitutive equation in which the viscosity depends on the local particle volume fraction and a diffusion equation that accounts for shear‐induced particle migration. Particle flux expressions used to obtain the diffusion equation are derived by simple scaling arguments. Predictions are made for the particle volume fraction and velocity fields for steady Couette and Poiseuille flow, and for transient start‐up of steady shear flow in a Couette apparatus. Particle concentrations for a monomodal suspension of polymethyl methacrylate spheres in a Newtonian solvent are measured by nuclear magnetic resonance (NMR) imaging in the Couette geometry for two particle sizes and volume fractions. The predictions agree remarkably well with the measurements for both transient and steady‐state experiments as well as for different particle sizes.

Often the adoption of component-based scientific software requires the developer to abandon comfortable practices and embrace an unfamiliar software methodology. OnRamp provides a mechanism for the developer to generate CCA components, through commented markup in their original software, and keep their familiar software development practices. The developer uses these annotations to identify which methods in their code belong to which Port interfaces, and which Ports belong to which CCA components. Taken by itself, the markup is sufficient to create a skeleton of components representing the exported functionality of the original code, but because the entire code is available to OnRamp the implementation can also be generated. The functionality of the original code is wrapped in annotation-specified components, exporting part or all of its original functionality. OnRamp provides a model for component software engineering that allows developers to improve their code with their familiar software and in their established software practices and also interoperate with external developers easily when the need arises.

The goal of this research was to combine theoretical and computational approaches to better understand the potential emergent behaviors of large-scale cyber systems, such as networks of {approx} 10{sup 6} computers. The scale and sophistication of modern computer software, hardware, and deployed networked systems have significantly exceeded the computational research community's ability to understand, model, and predict current and future behaviors. This predictive understanding, however, is critical to the development of new approaches for proactively designing new systems or enhancing existing systems with robustness to current and future cyber threats, including distributed malware such as botnets. We have developed preliminary theoretical and modeling capabilities that can ultimately answer questions such as: How would we reboot the Internet if it were taken down? Can we change network protocols to make them more secure without disrupting existing Internet connectivity and traffic flow? We have begun to address these issues by developing new capabilities for understanding and modeling Internet systems at scale. Specifically, we have addressed the need for scalable network simulation by carrying out emulations of a network with {approx} 10{sup 6} virtualized operating system instances on a high-performance computing cluster - a 'virtual Internet'. We have also explored mappings between previouslymore » studied emergent behaviors of complex systems and their potential cyber counterparts. Our results provide foundational capabilities for further research toward understanding the effects of complexity in cyber systems, to allow anticipating and thwarting hackers.« less

A method for assessing statistically quantifiable improvements in security for software vulnerabilities is presented. Drawing on concepts in complexity theory, undecidability, and previous work in high-reliability systems, we show that ensembles of similar implementations have statistical value even though each by itself is inscrutable. Research questions are identified that may allow practical application of these concepts.

This is meant as a place to put commentary on the whitepaper and is meant to be pretty much ad-hoc. Because the whitepaper describes a potential program in DOE ASCR and because it concerns many researchers in the field, these notes are meant to be extendable by anyone willing to put in the effort. Of course criticisms of the contents of the notes themselves are also welcome.

Experimental measurements of the shear stress and normal stresses in the viscometric flow of a well-characterized polyisobutylene/polybutene solution between a cone-and-plate and between parallel plates show the onset of an “anti-thixotropic” flow transition, which results in a time-dependent, apparent shear-thickening of the viscosity and first normal stress difference. Measurements of the critical conditions for the onset of the flow transition, made by systematically varying the plate separation in the parallel-plate geometry, demonstrate that this rotational instability is a function of the angular velocity in the flow and initially grows exponentially in time, in agreement with theoretical predictions for an Oldroyd-B model. However, experiments show that the disturbance which causes this flow transition is nonaxisymmetric, over-stable in time and subcritical in γ. Spectral analysis also shows that the nonlinear flow which ultimately develops is temporally aperiodic. These features of the flow transition do not agree with the existing theoretical analysis. The experimentally constructed flow-stability diagram shows that, for a given fluid, the instability occurs at a critical Deborah number which is insensitive to the specific geometrical parameters, temperature, and shear-rate of the flow. This interpretation of the elastic instability clarifies recent measurements of the flow stability for the Ml viscoelastic test fluid.

A phase‐space kinetic theory of dilute polymer solutions is developed to account for the effects of nonhomogeneous velocity and stress fields. The theory allows the configuration distribution function to depend on spatial location and explicitly treats the polymer molecule as an extended object in space. Constitutive equations for the mass flux vector and stress tensor are derived that predict polymer migration induced by stress gradients and nonuniform velocity gradients. In addition, the constitutive equation for stress contains a diffusive term in stress, and hence the model does not fall within the class of simple fluids. Simple shear flow between parallel plates is solved to illustrate the features of the constitutive equations. Asymptotic analysis and numerical calculations show the formation of boundary layers in stress, velocity gradient, and polymer concentration that arise near solid walls as a result of preferential orientation of the polymer molecules there. The thickness of these layers scale...

We report on the work done in the late-start LDRDUsing Emulation and Simulation toUnderstand the Large-Scale Behavior of the Internet. We describe the creation of a researchplatform that emulates many thousands of machines to be used for the study of large-scale inter-net behavior. We describe a proof-of-concept simple attack we performed in this environment.We describe the successful capture of a Storm bot and, from the study of the bot and furtherliterature search, establish large-scale aspects we seek to understand via emulation of Storm onour research platform in possible follow-on work. Finally, we discuss possible future work.3