Phosphatidylinositol 4-Kinase III Alpha Governs Cytoskeletal Organization for Invasiveness of Liver Cancer Cells.

BACKGROUND & AIMS: High expression of phosphatidylinositol 4-kinase III alpha (PI4KIIIα) correlates with poor survival rates in patients with hepatocellular carcinoma. In addition, hepatitis C virus (HCV) infections activate PI4KIIIα and contribute to hepatocellular carcinoma progression. We aimed at mechanistically understanding the impact of PI4KIIIα on the progression of liver cancer and the potential contribution of HCV in this process. METHODS: Several hepatic cell culture and mouse models were used to study the functional importance of PI4KIIIα on liver pathogenesis. Antibody arrays, gene silencing, and PI4KIIIα-specific inhibitor were applied to identify the involved signaling pathways. The contribution of HCV was examined by using HCV infection or overexpression of its nonstructural protein. RESULTS: High PI4KIIIα expression and/or activity induced cytoskeletal rearrangements via increased phosphorylation of paxillin and cofilin. This led to morphologic alterations and higher migratory and invasive properties of liver cancer cells. We further identified the liver-specific lipid kinase phosphatidylinositol 3-kinase C2 domain-containing subunit gamma (PIK3C2γ) working downstream of PI4KIIIα in regulation of the cytoskeleton. PIK3C2γ generates plasma membrane phosphatidylinositol 3,4-bisphosphate-enriched, invadopodia-like structures that regulate cytoskeletal reorganization by promoting Akt2 phosphorylation. CONCLUSIONS: PI4KIIIα regulates cytoskeleton organization via PIK3C2γ/Akt2/paxillin-cofilin to favor migration and invasion of liver cancer cells. These findings provide mechanistic insight into the contribution of PI4KIIIα and HCV to the progression of liver cancer and identify promising targets for therapeutic intervention.

Anti-merozoite antibodies induce natural killer cell effector function and are associated with immunity against malaria.

Natural killer (NK) cells are potent immune effectors that can be activated via antibody-mediated Fc receptor engagement. Using multiparameter flow cytometry, we found that NK cells degranulate and release IFN-γ upon stimulation with antibody-opsonized Plasmodium falciparum merozoites. Antibody-dependent NK (Ab-NK) activity was largely strain transcending and enhanced invasion inhibition into erythrocytes. Ab-NK was associated with the successful control of parasitemia after experimental malaria challenge in African adults. In an independent cohort study in children, Ab-NK increased with age, was boosted by concurrent P. falciparum infections, and was associated with a lower risk of clinical episodes of malaria. Nine of the 14 vaccine candidates tested induced Ab-NK, including some less well-characterized antigens: P41, P113, MSP11, RHOPH3, and Pf_11363200. These data highlight an important role of Ab-NK activity in immunity against malaria and provide a potential mechanism for evaluating vaccine candidates.

Simulation and control of dissolved air flotation and column froth flotation with simultaneous sedimentation.

Flotation is a separation process where particles or droplets are removed from a suspension with the aid of floating gas bubbles. Applications include dissolved air flotation (DAF) in industrial wastewater treatment and column froth flotation (CFF) in wastewater treatment and mineral processing. One-dimensional models of flotation have been limited to steady-state situations for half a century by means of the drift-flux theory. A newly developed dynamic one-dimensional model formulated in terms of partial differential equations can be used to predict the process of simultaneous flotation of bubbles and sedimentation of particles that are not attached to bubbles. The governing model is a pair of first-order conservation laws for the aggregate and solids volume fractions as functions of height and time. An analysis of nonlinear ingredients of the governing equations helps to identify desired steady-state operating conditions. These can be chosen by means of operating charts, which are diagrams that visualize regions of admissible values of the volumetric flows of the feed input and underflow outlet. This is detailed for the DAF thickening process. Dynamic simulations are obtained with a recently developed numerical method. Responses to control actions are demonstrated with scenarios in CFF and DAF.

Epstein-Barr virus and human papillomavirus serum antibodies define the viral status of nasopharyngeal carcinoma in a low endemic country.

Epstein-Barr virus (EBV) causes nasopharyngeal carcinoma (NPC) in endemic regions, where almost every tumor is EBV-positive. In Western populations, NPC is rare, and human papillomavirus infection (HPV) has been suggested as another viral cause. We validated multiplex serology with molecular tumor markers, to define EBV-positive, HPV-positive and EBV-/HPV-negative NPCs in the United Kingdom, and analyzed survival differences between those groups. Sera from NPC cases (n = 98) and age- and sex-matched controls (n = 142) from the Head and Neck 5000 clinical cohort study were analyzed. IgA and IgG serum antibodies against 13 EBV antigens were measured and compared with EBER in situ hybridization (EBER-ISH) data of 41 NPC tumors (29 EBER-ISH positive, 12 negative). IgG antibodies to EBV LF2 correctly diagnosed EBV-positive NPCs in 28 of 29 cases, while all EBER-ISH negative NPCs were seronegative to LF2 IgG (specificity = 100%, sensitivity = 97%). HPV early antigen serology was compared to HPV molecular markers (p16 expression, HPV DNA and RNA) available for 41 NPCs (13 positive, 28 negative). Serology matched molecular HPV markers in all but one case (specificity = 100%, sensitivity = 92%). EBV and HPV infections were mutually exclusive. Overall, 67% of the analyzed NPCs were defined as EBV-positive, 18% as HPV-positive and 14% as EBV/HPV-negative. There was no statistical evidence of a difference in survival between the three groups. These data provide evidence that both, EBV-positive and HPV-positive NPCs are present in a low incidence country, and that EBV and HPV serum antibodies correlate with the viral status of the tumor.

Analysis of photobioreactors in series.

A photobioreactor (PBR) contains microalgae which under illumination consume carbon dioxide and substrate dissolved in water, and produce oxygen. The process is used in water recovery resource facilities with a continuous flow of wastewaster through the PBR. With several PBRs in series the reduction of substrate can be improved. This paper contains a thorough analysis of a model of PBRs in series, where each PBR is modelled with a system of three ordinary differential equations for the concentrations of dissolved substrate and biomass (algae), and the internal cell quota of substrate to biomass. Each PBR has a certain volume and irradiation. The absorption rate of substrate into the cells is modelled with Monod kinetics, whereas the biomass growth rate is modelled with Droop kinetics, in which both a minimum and a maximum internal cell quota are assumed. The main result is that the model has a unique stable steady-state solution with algae in all PBRs. Another stable steady-state solution is the wash-out solution with no algae in the system. Other steady-state solutions are combinations of these two with no algae in some of the first PBRs and algae in the rest of the PBRs in the series. Conditions on the illumination, volumetric flow and volumes of the PBRs are given for the respective solution. Numerical solutions illustrate the theoretical results and indicate further properties.

Concentration-driven models revisited: towards a unified framework to model settling tanks in water resource recovery facilities.

A new perspective on the modelling of settling behaviour in water resource recovery facilities is introduced. The ultimate goal is to describe in a unified way the processes taking place both in primary settling tanks (PSTs) and secondary settling tanks (SSTs) for a more detailed operation and control. First, experimental evidence is provided, pointing out distributed particle properties (such as size, shape, density, porosity, and flocculation state) as an important common source of distributed settling behaviour in different settling unit processes and throughout different settling regimes (discrete, hindered and compression settling). Subsequently, a unified model framework that considers several particle classes is proposed in order to describe distributions in settling behaviour as well as the effect of variations in particle properties on the settling process. The result is a set of partial differential equations (PDEs) that are valid from dilute concentrations, where they correspond to discrete settling, to concentrated suspensions, where they correspond to compression settling. Consequently, these PDEs model both PSTs and SSTs.

On constitutive functions for hindered settling velocity in 1-D settler models: Selection of appropriate model structure.

Advanced 1-D models for Secondary Settling Tanks (SSTs) explicitly account for several phenomena that influence the settling process (such as hindered settling and compression settling). For each of these phenomena a valid mathematical expression needs to be selected and its parameters calibrated to obtain a model that can be used for operation and control. This is, however, a challenging task as these phenomena may occur simultaneously. Therefore, the presented work evaluates several available expressions for hindered settling based on long-term batch settling data. Specific attention is paid to the behaviour of these hindered settling functions in the compression region in order to evaluate how the modelling of sludge compression is influenced by the choice of a certain hindered settling function. The analysis shows that the exponential hindered settling forms, which are most commonly used in traditional SST models, not only account for hindered settling but partly lump other phenomena (compression) as well. This makes them unsuitable for advanced 1-D models that explicitly include each phenomenon in a modular way. A power-law function is shown to be more appropriate to describe the hindered settling velocity in advanced 1-D SST models.

Efficient simulations of tubulin-driven axonal growth.

This work concerns efficient and reliable numerical simulations of the dynamic behaviour of a moving-boundary model for tubulin-driven axonal growth. The model is nonlinear and consists of a coupled set of a partial differential equation (PDE) and two ordinary differential equations. The PDE is defined on a computational domain with a moving boundary, which is part of the solution. Numerical simulations based on standard explicit time-stepping methods are too time consuming due to the small time steps required for numerical stability. On the other hand standard implicit schemes are too complex due to the nonlinear equations that needs to be solved in each step. Instead, we propose to use the Peaceman-Rachford splitting scheme combined with temporal and spatial scalings of the model. Simulations based on this scheme have shown to be efficient, accurate, and reliable which makes it possible to evaluate the model, e.g. its dependency on biological and physical model parameters. These evaluations show among other things that the initial axon growth is very fast, that the active transport is the dominant reason over diffusion for the growth velocity, and that the polymerization rate in the growth cone does not affect the final axon length.

On time discretizations for the simulation of the batch settling-compression process in one dimension.

The main purpose of the recently introduced Bürger-Diehl simulation model for secondary settling tanks was to resolve spatial discretization problems when both hindered settling and the phenomena of compression and dispersion are included. Straightforward time integration unfortunately means long computational times. The next step in the development is to introduce and investigate time-integration methods for more efficient simulations, but where other aspects such as implementation complexity and robustness are equally considered. This is done for batch settling simulations. The key findings are partly a new time-discretization method and partly its comparison with other specially tailored and standard methods. Several advantages and disadvantages for each method are given. One conclusion is that the new linearly implicit method is easier to implement than another one (semi-implicit method), but less efficient based on two types of batch sedimentation tests.

A consistent modelling methodology for secondary settling tanks: a reliable numerical method.

The consistent modelling methodology for secondary settling tanks (SSTs) leads to a partial differential equation (PDE) of nonlinear convection-diffusion type as a one-dimensional model for the solids concentration as a function of depth and time. This PDE includes a flux that depends discontinuously on spatial position modelling hindered settling and bulk flows, a singular source term describing the feed mechanism, a degenerating term accounting for sediment compressibility, and a dispersion term for turbulence. In addition, the solution itself is discontinuous. A consistent, reliable and robust numerical method that properly handles these difficulties is presented. Many constitutive relations for hindered settling, compression and dispersion can be used within the model, allowing the user to switch on and off effects of interest depending on the modelling goal as well as investigate the suitability of certain constitutive expressions. Simulations show the effect of the dispersion term on effluent suspended solids and total sludge mass in the SST. The focus is on correct implementation whereas calibration and validation are not pursued.

Fundamental nonlinearities of the reactor-settler interaction in the activated sludge process.

The activated sludge process can be modelled by ordinary and partial differential equations for the biological reactors and secondary settlers, respectively. Because of the complexity of such a system, simulation models are most often used to investigate them. However, simulation models cannot give general rules on how to control a complex nonlinear process. For a reduced-order model with only two components, soluble substrate and particulate biomass, general results on steady-state solutions have recently been obtained, such as existence, uniqueness and stability of solutions. The aim of the present paper is to utilize those results to formulate some implications of practical importance. In particular, strategies are described for the manual control of the effluent substrate concentration subject to the constraint that the settler is maintained in normal operation (with a sludge blanket in the thickening zone) in steady state. Such strategies contain how the two control parameters, the recycle and waste volumetric flow ratios, should be chosen for any (steady-state) values of the input variables.

Theory of EPR lineshape in samples concentrated in paramagnetic spins: effect of enhanced internal magnetic field on high-field high-frequency (HFHF) EPR lineshape.

A theoretical treatment is provided for the calculation of EPR (electron paramagnetic resonance) lineshape as affected by interactions with paramagnetic ions in the vicinity. The internal fields seen by the various paramagnetic ions due to interactions with paramagnetic ions in their vicinity, as well as the resulting lineshapes, become quite significant at high magnetic fields required in high-frequency (HFHF) EPR. The resulting EPR signals for the various ions are therefore characterized by different g-shifts and lineshapes, so that the overall EPR lineshape, which is an overlap of these, becomes distorted, or even split in HFHF EPR, from that observed at lower frequencies. The observed EPR lineshapes in MnSO(4)⋅H(2)O powder and K(3)CrO(8) single-crystal samples have been simulated here taking into account g-shifts and modified lineshapes. These simulations show that in these samples, concentrated in paramagnetic spins, the position and lineshapes of EPR signals are significantly modified in HFHF EPR involving very high magnetic fields.

A consistent modelling methodology for secondary settling tanks in wastewater treatment.

The aim of this contribution is partly to build consensus on a consistent modelling methodology (CMM) of complex real processes in wastewater treatment by combining classical concepts with results from applied mathematics, and partly to apply it to the clarification-thickening process in the secondary settling tank. In the CMM, the real process should be approximated by a mathematical model (process model; ordinary or partial differential equation (ODE or PDE)), which in turn is approximated by a simulation model (numerical method) implemented on a computer. These steps have often not been carried out in a correct way. The secondary settling tank was chosen as a case since this is one of the most complex processes in a wastewater treatment plant and simulation models developed decades ago have no guarantee of satisfying fundamental mathematical and physical properties. Nevertheless, such methods are still used in commercial tools to date. This particularly becomes of interest as the state-of-the-art practice is moving towards plant-wide modelling. Then all submodels interact and errors propagate through the model and severely hamper any calibration effort and, hence, the predictive purpose of the model. The CMM is described by applying it first to a simple conversion process in the biological reactor yielding an ODE solver, and then to the solid-liquid separation in the secondary settling tank, yielding a PDE solver. Time has come to incorporate established mathematical techniques into environmental engineering, and wastewater treatment modelling in particular, and to use proven reliable and consistent simulation models.

A multifrequency EPR study of Fe2+ and Mn2+ ions in a ZnSiF(6).6H2O single crystal at liquid-helium temperatures.

A liquid-helium temperature study of Fe2+ and Mn2+ ions has been carried out on a single crystal of Fe2+-doped ZnSiF(6).6H2O at 5-35K at 170, 222.4 and 333.2G Hz. The spectra are found to be an overlap of two magnetically inequivalent Fe2+ ions, as well as that of an Mn2+ ion. From the simulation of the EPR line positions for the Fe2+ (d6, S=2) ion the spin-Hamiltonian parameters were estimated for the two inequivalent Fe2+ ions at the various temperatures. From the relative intensities of lines the absolute sign of the fine-structure parameters have been estimated. In addition, the fine-structure and hyperfine-structure spin-Hamiltonian parameters for the Mn2+ ion, present as impurity at interstitial sites, were estimated from the hyperfine allowed and forbidden line positions. The particular virtues of such a single-crystal study vs. that on powders are noted.

The solids-flux theory--confirmation and extension by using partial differential equations.

The solids-flux theory has been used for half a century as a tool for estimating concentration and fluxes in the design and operation of secondary settling tanks during stationary conditions. The flux theory means that the conservation of mass is used in one dimension together with the batch-settling flux function according to the Kynch assumption. The flux theory results correspond to stationary solutions of a partial differential equation, a conservation law, with discontinuous coefficients modelling the continuous-sedimentation process in one dimension. The mathematical analysis of such an equation is intricate, partly since it cannot be interpreted in the classical sense. Recent results, however, make it possible to partly confirm and extend the previous flux theory statements, partly draw new conclusions also on the dynamic behaviour and the possibilities and limitations for control. We use here a single example of an ideal settling tank and a given batch-settling flux in a whole series of calculations. The mathematical results are adapted towards the application and many of them are conveniently presented in terms of operating charts.