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Sample records for 2-d flow cell

  1. Flow past 2-D Hemispherical Rigid Canopies

    NASA Astrophysics Data System (ADS)

    Carnasciali, Maria-Isabel

    2013-11-01

    The flow past a 2-dimensional rigid hemispherical shape is investigated using PIV. Flow field measurements and images were generated with the use of a Thermoflow® apparatus. Results of this study are compared to prior work (APS DFD 2012 Session E9.00003) which employed CFD to investigate the flow in the near wake of hemispherical parachutes. The various sized gaps/open areas were positioned at distinct locations. The work presented here is part of a larger research project to investigate flow fields in deceleration devices and parachutes. Understanding the pitch-stability of parachutes is essential for accurate design and implementation of these deceleration devices but they present a difficult system to analyze. The flexibility of the parachute fabric results in large variations in the parachute geometry leading to complex fluid-structure interactions. Such flow, combined with flow through gaps and open areas, has been postulated to shed alternating vortices causing pitching/oscillations of the canopy. The results presented provide some insight into which geometric features affect vortex shedding and may enable the redesign of the baseline parachute to minimize instabilities.

  2. Mean flow and anisotropic cascades in decaying 2D turbulence

    NASA Astrophysics Data System (ADS)

    Liu, Chien-Chia; Cerbus, Rory; Gioia, Gustavo; Chakraborty, Pinaki

    2015-11-01

    Many large-scale atmospheric and oceanic flows are decaying 2D turbulent flows embedded in a non-uniform mean flow. Despite its importance for large-scale weather systems, the affect of non-uniform mean flows on decaying 2D turbulence remains unknown. In the absence of mean flow it is well known that decaying 2D turbulent flows exhibit the enstrophy cascade. More generally, for any 2D turbulent flow, all computational, experimental and field data amassed to date indicate that the spectrum of longitudinal and transverse velocity fluctuations correspond to the same cascade, signifying isotropy of cascades. Here we report experiments on decaying 2D turbulence in soap films with a non-uniform mean flow. We find that the flow transitions from the usual isotropic enstrophy cascade to a series of unusual and, to our knowledge, never before observed or predicted, anisotropic cascades where the longitudinal and transverse spectra are mutually independent. We discuss implications of our results for decaying geophysical turbulence.

  3. Flow transitions in a 2D directional solidification model

    NASA Technical Reports Server (NTRS)

    Larroude, Philippe; Ouazzani, Jalil; Alexander, J. Iwan D.

    1992-01-01

    Flow transitions in a Two Dimensional (2D) model of crystal growth were examined using the Bridgman-Stockbarger me thod. Using a pseudo-spectral Chebyshev collocation method, the governing equations yield solutions which exhibit a symmetry breaking flow tansition and oscillatory behavior indicative of a Hopf bifurcation at higher values of Ra. The results are discussed from fluid dynamic viewpoint, and broader implications for process models are also addressed.

  4. Advecting Procedural Textures for 2D Flow Animation

    NASA Technical Reports Server (NTRS)

    Kao, David; Pang, Alex; Moran, Pat (Technical Monitor)

    2001-01-01

    This paper proposes the use of specially generated 3D procedural textures for visualizing steady state 2D flow fields. We use the flow field to advect and animate the texture over time. However, using standard texture advection techniques and arbitrary textures will introduce some undesirable effects such as: (a) expanding texture from a critical source point, (b) streaking pattern from the boundary of the flowfield, (c) crowding of advected textures near an attracting spiral or sink, and (d) absent or lack of textures in some regions of the flow. This paper proposes a number of strategies to solve these problems. We demonstrate how the technique works using both synthetic data and computational fluid dynamics data.

  5. 2-D Model for Normal and Sickle Cell Blood Microcirculation

    NASA Astrophysics Data System (ADS)

    Tekleab, Yonatan; Harris, Wesley

    2011-11-01

    Sickle cell disease (SCD) is a genetic disorder that alters the red blood cell (RBC) structure and function such that hemoglobin (Hb) cannot effectively bind and release oxygen. Previous computational models have been designed to study the microcirculation for insight into blood disorders such as SCD. Our novel 2-D computational model represents a fast, time efficient method developed to analyze flow dynamics, O2 diffusion, and cell deformation in the microcirculation. The model uses a finite difference, Crank-Nicholson scheme to compute the flow and O2 concentration, and the level set computational method to advect the RBC membrane on a staggered grid. Several sets of initial and boundary conditions were tested. Simulation data indicate a few parameters to be significant in the perturbation of the blood flow and O2 concentration profiles. Specifically, the Hill coefficient, arterial O2 partial pressure, O2 partial pressure at 50% Hb saturation, and cell membrane stiffness are significant factors. Results were found to be consistent with those of Le Floch [2010] and Secomb [2006].

  6. Enhancement of biomixing by swimming cells in 2D films

    NASA Astrophysics Data System (ADS)

    Gollub, Jerry; Kurtuldu, Huseyin; Guasto, Jeffrey; Johnson, Karl

    2011-11-01

    Fluid mixing in active suspensions of microorganisms is important to ecological phenomena and shows surprising statistical behavior. We investigate the mixing produced by swimming unicellular algal cells (Chlamydomonas) in quasi-2D films by tracking the motions of cells and of microscopic passive tracer particles advected by the fluid. The reduced spatial dimension of the system leads to long-range flows and a surprisingly strong dependence of tracer transport on the swimmer concentration. The mean square displacements are well described by a stochastic Langevin model, with an effective diffusion coefficient D growing as the 3/2 power of the swimmer concentration, due to the interaction of tracer particles with multiple swimmers. We also discuss the anomalous probability distributions of tracer displacements, which become Gaussian at high concentration, but show strong power-law tails at low concentration. Supported by NSF Grant DMR-0803153.

  7. CFD code comparison for 2D airfoil flows

    NASA Astrophysics Data System (ADS)

    Sørensen, Niels N.; Méndez, B.; Muñoz, A.; Sieros, G.; Jost, E.; Lutz, T.; Papadakis, G.; Voutsinas, S.; Barakos, G. N.; Colonia, S.; Baldacchino, D.; Baptista, C.; Ferreira, C.

    2016-09-01

    The current paper presents the effort, in the EU AVATAR project, to establish the necessary requirements to obtain consistent lift over drag ratios among seven CFD codes. The flow around a 2D airfoil case is studied, for both transitional and fully turbulent conditions at Reynolds numbers of 3 × 106 and 15 × 106. The necessary grid resolution, domain size, and iterative convergence criteria to have consistent results are discussed, and suggestions are given for best practice. For the fully turbulent results four out of seven codes provide consistent results. For the laminar-turbulent transitional results only three out of seven provided results, and the agreement is generally lower than for the fully turbulent case.

  8. Position control using 2D-to-2D feature correspondences in vision guided cell micromanipulation.

    PubMed

    Zhang, Yanliang; Han, Mingli; Shee, Cheng Yap; Ang, Wei Tech

    2007-01-01

    Conventional camera calibration that utilizes the extrinsic and intrinsic parameters of the camera and the objects has certain limitations for micro-level cell operations due to the presence of hardware deviations and external disturbances during the experimental process, thereby invalidating the extrinsic parameters. This invalidation is often neglected in macro-world visual servoing and affects the visual image processing quality, causing deviation from the desired position in micro-level cell operations. To increase the success rate of vision guided biological micromanipulations, a novel algorithm monitoring the changing image pattern of the manipulators including the injection micropipette and cell holder is designed and implemented based on 2 dimensional (2D)-to 2D feature correspondences and can adjust the manipulator and perform position control simultaneously. When any deviation is found, the manipulator is retracted to the initial focusing plane before continuing the operation.

  9. Flow-induced 2D protein crystallization: characterization of the coupled interfacial and bulk flows.

    PubMed

    Young, James E; Posada, David; Lopez, Juan M; Hirsa, Amir H

    2015-05-14

    Two-dimensional crystallization of the protein streptavidin, crystallizing below a biotinylated lipid film spread on a quiescent air-water interface is a well studied phenomenon. More recently, 2D crystallization induced by a shearing interfacial flow has been observed at film surface pressures significantly lower than those required in a quiescent system. Here, we quantify the interfacial and bulk flow associated with 2D protein crystallization through numerical modeling of the flow along with a Newtonian surface model. Experiments were conducted over a wide range of conditions resulting in a state diagram delineating the flow strength required to induce crystals for various surface pressures. Through measurements of the velocity profile at the air-water interface, we found that even in the cases where crystals are formed, the macroscopic flow at the interface is well described by the Newtonian model. However, the results show that even in the absence of any protein in the system, the viscous response of the biotinylated lipid film is complicated and strongly dependent on the strength of the flow. This observation suggests that the insoluble lipid film plays a key role in flow-induced 2D protein crystallization.

  10. NKG2D ligands mediate immunosurveillance of senescent cells.

    PubMed

    Sagiv, Adi; Burton, Dominick G A; Moshayev, Zhana; Vadai, Ezra; Wensveen, Felix; Ben-Dor, Shifra; Golani, Ofra; Polic, Bojan; Krizhanovsky, Valery

    2016-02-01

    Cellular senescence is a stress response mechanism that limits tumorigenesis and tissue damage. Induction of cellular senescence commonly coincides with an immunogenic phenotype that promotes self-elimination by components of the immune system, thereby facilitating tumor suppression and limiting excess fibrosis during wound repair. The mechanisms by which senescent cells regulate their immune surveillance are not completely understood. Here we show that ligands of an activating Natural Killer (NK) cell receptor (NKG2D), MICA and ULBP2 are consistently up-regulated following induction of replicative senescence, oncogene-induced senescence and DNA damage - induced senescence. MICA and ULBP2 proteins are necessary for efficient NK-mediated cytotoxicity towards senescent fibroblasts. The mechanisms regulating the initial expression of NKG2D ligands in senescent cells are dependent on a DNA damage response, whilst continuous expression of these ligands is regulated by the ERK signaling pathway. In liver fibrosis, the accumulation of senescent activated stellate cells is increased in mice lacking NKG2D receptor leading to increased fibrosis. Overall, our results provide new insights into the mechanisms regulating the expression of immune ligands in senescent cells and reveal the importance of NKG2D receptor-ligand interaction in protecting against liver fibrosis. PMID:26878797

  11. Tetramethypyrazine inhibits renal cell carcinoma cells through inhibition of NKG2D signaling pathways.

    PubMed

    Luan, Yun; Liu, Junli; Liu, Xiaoli; Xue, Xia; Kong, Feng; Sun, Chao; Wang, Jue; Liu, Ling; Jia, Hongying

    2016-10-01

    Tetramethypyrazine (TMP), one of the active compounds extracted from the traditional Chinese medicinal herb (Chuanxiong), has been verified as an anticancer compound against several types of cancer. However, understanding of the molecular mechanisms have not been fully elucidated. In the present study, the anticancer efficacy of TMP was investigated in human clear cell renal cell carcinoma (ccRCC) cells. We showed that TMP significantly inhibited ccRCC cell viability, proliferation, apoptosis, invasion and migration through the methods of MTT, flow cytometry, wound healing and transwell assays. Furthermore, reverse transcription polymerase chain reaction (RT-PCR), western blotting and immunofluorescence results demonstrated TMP upregulation of the expression of NKG2D ligands (NKG2DLs) MHC class I chain-related molecules A and B (MICA/B) and epithelial cell expression marker of E-cadherin, and downregulation of mesenchymal cell expression markers of vimentin and fibronectin. Taken together, the inhibition of TMP on ccRCC cells might be mediated via inhibition of NKG2D related signaling pathway to further suppress epithelial-mesenchymal transition (EMT) progression. The binding of NKG2D to its ligands activates NK cells, giving the rationale for studies on the utilization of TMP as a potential cancer therapeutic compound to increase NK cells-mediated cytotoxicity against high MICA/B expression in cancer cells. PMID:27633040

  12. Cryogenic cavitating flow in 2D laval nozzle

    NASA Astrophysics Data System (ADS)

    Tani, Naoki; Nagashima, Toshio

    2003-05-01

    Cavitation is one of the troublesome problems in rocket turbo pumps, and since most of high-efficiency rocket propellants are cryogenic fluids, so called “thermodynamic effect” becomes more evident than in water. In the present study, numerical and experimental study of liquid nitrogen cavitation in 2D Laval nozzle was carried out, so that the influence of thermodynamic effect was examined. It was revealed that temperature and cavitation have strong inter-relationship with each other in thermo-sensitive cryogenic fluids.

  13. Improvement of a 2D numerical model of lava flows

    NASA Astrophysics Data System (ADS)

    Ishimine, Y.

    2013-12-01

    I propose an improved procedure that reduces an improper dependence of lava flow directions on the orientation of Digital Elevation Model (DEM) in two-dimensional simulations based on Ishihara et al. (in Lava Flows and Domes, Fink, JH eds., 1990). The numerical model for lava flow simulations proposed by Ishihara et al. (1990) is based on two-dimensional shallow water model combined with a constitutive equation for a Bingham fluid. It is simple but useful because it properly reproduces distributions of actual lava flows. Thus, it has been regarded as one of pioneer work of numerical simulations of lava flows and it is still now widely used in practical hazard prediction map for civil defense officials in Japan. However, the model include an improper dependence of lava flow directions on the orientation of DEM because the model separately assigns the condition for the lava flow to stop due to yield stress for each of two orthogonal axes of rectangular calculating grid based on DEM. This procedure brings a diamond-shaped distribution as shown in Fig. 1 when calculating a lava flow supplied from a point source on a virtual flat plane although the distribution should be circle-shaped. To improve the drawback, I proposed a modified procedure that uses the absolute value of yield stress derived from both components of two orthogonal directions of the slope steepness to assign the condition for lava flows to stop. This brings a better result as shown in Fig. 2. Fig. 1. (a) Contour plots calculated with the original model of Ishihara et al. (1990). (b) Contour plots calculated with a proposed model.

  14. Photonic crystal based 2D integrating cell for sensing applications

    NASA Astrophysics Data System (ADS)

    Fohrmann, Lena Simone; Petrov, Alexander Y.; Sommer, Gerrit; Krauss, Thomas; Eich, Manfred

    2016-04-01

    We present a concept of a silicon slab based 2D integrating cell where photonic crystal (PhC) reflectors are used in order to confine light in a two-dimensional area to acquire a long propagation length. The evanescent field of the guided wave can be used for sensing applications. We use FDTD simulations to investigate the dependence of the reflectivity of photonic crystal mirrors with a hexagonal lattice. The reflectivity in ΓM direction demonstrates reduced vertical losses compared to the ΓK direction and can be further improved by adiabatically tapering the hole radii of the photonic crystal. A small hexagonal 2D integrating cell was studied with PhC boundaries oriented in ΓM and ΓK direction. It is shown that average reflectivities of 99% can be obtained in a rectangular 2D cell with optimized reflector design, limited only by residual vertical scattering losses at the PhC boundary. This reflectivity is already comparable to the best metallic reflectors.

  15. COYOTE: A computer program for 2-D reactive flow simulations

    SciTech Connect

    Cloutman, L.D.

    1990-04-01

    We describe the numerical algorithm used in the COYOTE two- dimensional, transient, Eulerian hydrodynamics program for reactive flows. The program has a variety of options that provide capabilities for a wide range of applications, and it is designed to be robust and relatively easy to use while maintaining adequate accuracy and efficiency to solve realistic problems. It is based on the ICE method, and it includes a general species and chemical reaction network for simulating reactive flows. It also includes swirl, turbulence transport models, and a nonuniform mesh capability. We describe several applications of the program. 33 refs., 4 figs.

  16. 2D VARIABLY SATURATED FLOWS: PHYSICAL SCALING AND BAYESIAN ESTIMATION

    EPA Science Inventory

    A novel dimensionless formulation for water flow in two-dimensional variably saturated media is presented. It shows that scaling physical systems requires conservation of the ratio between capillary forces and gravity forces. A direct result of this finding is that for two phys...

  17. 2D Mixed Convection Thermal Incompressible Viscous Flows

    NASA Astrophysics Data System (ADS)

    Bermudez, Blanca; Nicolas, Alfredo

    2005-11-01

    Mixed convection thermal incomprressible viscous fluid flows in rectangular cavities are presented. These kind of flows may be governed by the time-dependent Boussinesq approximation in terms of the stream function-vorticity variables formulation. The results are obtained with a simple numerical scheme based mainly on a fixed point iterative process applied to the non-linear system of elliptic equations that is obtained after a second order time discretization. Numerical experiments are reported for the problem of a cavity with fluid boundary motion on the top. Some results correspond to validation examples and others, to the best of our knowledge, correspond to new results. To show that the new results are correct, a mesh size and time independence studies are carried out, and the acceptable errors are measured point-wise. For the optimal mesh size and time step the final times when the steady state is reached, as solution from the unsteady problem, are reported; it should be seen that they are larger than the ones for natural convection which, physically speaking, show the agreement that mixed convection flows are more active than those of natural convection due to the fluid boundary motion on the top of the cavity. The flow parameters are: the Reynolds number, the Grashof number and the aspect ratio.

  18. Lagrangian statistics and flow topology in forced 2-D turbulence

    SciTech Connect

    Kadoch, B.; Del-Castillo-Negrete, Diego B; Bos, W.J.T.; Schneider, Kai

    2011-01-01

    A study of the relationship between Lagrangian statistics and flow topology in fluid turbulence is presented. The topology is characterized using the Weiss criterion, which provides a conceptually simple tool to partition the flow into topologically different regions: elliptic (vortex dominated), hyperbolic (deformation dominated), and intermediate (turbulent background). The flow corresponds to forced two-dimensional Navier-Stokes turbulence in doubly periodic and circular bounded domains, the latter with no-slip boundary conditions. In the double periodic domain, the probability density function (pdf) of the Weiss field exhibits a negative skewness consistent with the fact that in periodic domains the flow is dominated by coherent vortex structures. On the other hand, in the circular domain, the elliptic and hyperbolic regions seem to be statistically similar. We follow a Lagrangian approach and obtain the statistics by tracking large ensembles of passively advected tracers. The pdfs of residence time in the topologically different regions are computed introducing the Lagrangian Weiss field, i.e., the Weiss field computed along the particles' trajectories. In elliptic and hyperbolic regions, the pdfs of the residence time have self-similar algebraic decaying tails. In contrast, in the intermediate regions the pdf has exponential decaying tails. The conditional pdfs (with respect to the flow topology) of the Lagrangian velocity exhibit Gaussian-like behavior in the periodic and in the bounded domains. In contrast to the freely decaying turbulence case, the conditional pdfs of the Lagrangian acceleration in forced turbulence show a comparable level of intermittency in both the periodic and the bounded domains. The conditional pdfs of the Lagrangian curvature are characterized, in all cases, by self-similar power-law behavior with a decay exponent of order - 2.

  19. Controlling cell growth with tailorable 2D nanoholes arrays.

    PubMed

    Fragal, Vanessa H; Cellet, Thelma Sley P; Fragal, Elizângela H; Pereira, Guilherme M; Garcia, Francielle P; Nakamura, Celso V; Asefa, Tewodros; Rubira, Adley F; Silva, Rafael

    2016-03-15

    A facile and reproducible route that can lead to two-dimensional arrays of nanopores in thin polymer films is demonstrated. The formation of the pores in the polymer films involves breath figure phenomenon and occurs during the film deposition by spin coating. The formation of nanoporous thin films takes only few seconds, and the method does not require complex equipment or expensive chemicals. This method also constitutes a straightforward approach to control the size of the pores formed in thin films. Besides allowing control over the average pore size of the porous films, the use of dynamic deposition with the breath figure phenomenon causes the reduction in the pore size to nanometer scale. The nanoporous arrays obtained by the breath figure are applied as substrates for cell growth, and the effect of their nanopore size on cell growth was evaluated. Notably, it is found that cell viability is related to pore size, where 2D nanoporous structure is more beneficial for cell culture than 2D microporous structures. The change in the average pore size of the polymer films from 1.22 μm to 346 nm results in a threefold increase in cell viability. PMID:26722796

  20. Flow Solver for Incompressible 2-D Drive Cavity

    NASA Technical Reports Server (NTRS)

    Kalb, Virginia L.

    2008-01-01

    This software solves the Navier-Stokes equations for the incompressible driven cavity flow problem. The code uses second-order finite differencing on a staggered grid using the Chorin projection method. The resulting intermediate Poisson equation is efficiently solved using the fast Fourier transform. Time stepping is done using fourth-order Runge-Kutta for stability at high Reynolds numbers. Features include check-pointing, periodic field snapshots, ongoing reporting of kinetic energy and changes between time steps, time histories at selected points, and optional streakline generation.

  1. In-Cell Protein Structures from 2D NMR Experiments.

    PubMed

    Müntener, Thomas; Häussinger, Daniel; Selenko, Philipp; Theillet, Francois-Xavier

    2016-07-21

    In-cell NMR spectroscopy provides atomic resolution insights into the structural properties of proteins in cells, but it is rarely used to solve entire protein structures de novo. Here, we introduce a paramagnetic lanthanide-tag to simultaneously measure protein pseudocontact shifts (PCSs) and residual dipolar couplings (RDCs) to be used as input for structure calculation routines within the Rosetta program. We employ this approach to determine the structure of the protein G B1 domain (GB1) in intact Xenopus laevis oocytes from a single set of 2D in-cell NMR experiments. Specifically, we derive well-defined GB1 ensembles from low concentration in-cell NMR samples (∼50 μM) measured at moderate magnetic field strengths (600 MHz), thus offering an easily accessible alternative for determining intracellular protein structures. PMID:27379949

  2. An Integrative Model of Excitation Driven Fluid Flow in a 2D Uterine Channel

    NASA Astrophysics Data System (ADS)

    Maggio, Charles; Fauci, Lisa; Chrispell, John

    2009-11-01

    We present a model of intra-uterine fluid flow in a sagittal cross-section of the uterus by inducing peristalsis in a 2D channel. This is an integrative multiscale computational model that takes as input fluid viscosity, passive tissue properties of the uterine channel and a prescribed wave of membrane depolarization. This voltage pulse is coupled to a model of calcium dynamics inside a uterine smooth muscle cell, which in turn drives a kinetic model of myosin phosphorylation governing contractile muscle forces. Using the immersed boundary method, these muscle forces are communicated to a fluid domain to simulate the contractions which occur in a human uterus. An analysis of the effects of model parameters on the flow properties and emergent geometry of the peristaltic channel will be presented.

  3. Numerical Simulation of Supersonic Compression Corners and Hypersonic Inlet Flows Using the RPLUS2D Code

    NASA Technical Reports Server (NTRS)

    Kapoor, Kamlesh; Anderson, Bernhard H.; Shaw, Robert J.

    1994-01-01

    A two-dimensional computational code, PRLUS2D, which was developed for the reactive propulsive flows of ramjets and scramjets, was validated for two-dimensional shock-wave/turbulent-boundary-layer interactions. The problem of compression corners at supersonic speeds was solved using the RPLUS2D code. To validate the RPLUS2D code for hypersonic speeds, it was applied to a realistic hypersonic inlet geometry. Both the Baldwin-Lomax and the Chien two-equation turbulence models were used. Computational results showed that the RPLUS2D code compared very well with experimentally obtained data for supersonic compression corner flows, except in the case of large separated flows resulting from the interactions between the shock wave and turbulent boundary layer. The computational results compared well with the experiment results in a hypersonic NASA P8 inlet case, with the Chien two-equation turbulence model performing better than the Baldwin-Lomax model.

  4. Quantitative and functional analysis of CD69(+) NKG2D(+) T regulatory cells in healthy subjects.

    PubMed

    Vitales-Noyola, M; Doníz-Padilla, L; Álvarez-Quiroga, C; Monsiváis-Urenda, A; Portillo-Salazar, H; González-Amaro, R

    2015-07-01

    T regulatory (Treg) cells have a key role in immune homeostasis and the pathogenesis of chronic inflammatory and autoimmune diseases. CD69 is an early leukocyte activation molecule that under steady state conditions is detected in a small proportion of lymphocytes in peripheral blood and lymphoid tissues. Although it has been reported that a subset of CD69(+) T cells behaves as Treg lymphocytes, the possible relationship between CD69(+) Treg cells and CD4(+)NKG2D(+) T lymphocytes, which also exert immunosuppressive activity, has not been explored. In this study, we analyzed the expression of CD69 and NKG2D by T lymphocytes from the peripheral blood of twenty-five healthy subjects by multi-parametric flow cytometry analysis, and their suppressive activity by an assay of inhibition of lymphocyte activation (CD40L expression) and proliferation (carboxyfluorescein partition assay). We found a very small percentage of CD4(+)CD69(+)NKG2D(+) T cells (median 0.002%, Q1-Q3, 0.001-0.004%), which also expressed TGF-β (Latency Associated Peptide or LAP) and IL-10, in all samples analyzed. These cells exerted an important in vitro suppressive effect on both activation and proliferation of T effector cells. Our data suggest that at very small numbers, CD4(+)CD69(+)NKG2D(+) lymphocytes seem to exert a relevant functional immune-regulatory role in healthy subjects.

  5. Effects of NKG2D haplotypes on the cell-surface expression of NKG2D protein on natural killer and CD8 T cells of peripheral blood among atomic-bomb survivors.

    PubMed

    Imai, Kazue; Hayashi, Tomonori; Yamaoka, Mika; Kajimura, Junko; Yoshida, Kengo; Kusunoki, Yoichiro; Nakachi, Kei

    2012-06-01

    NKG2D is a primary activating receptor that triggers cell-mediated cytotoxicity in NK cells against tumor and virus-infected cells. We previously identified the NKG2D haplotypes in the natural killer gene complex region on chromosome 12p. Two major haplotype alleles, LNK1 and HNK1, were closely related to low and high natural cytotoxic activity phenotypes, respectively. Furthermore, the haplotype of HNK1/HNK1 has revealed a decreased risk of cancer compared with LNK1/LNK1. In the present study, using flow cytometry, we evaluated the functional effects of NKG2D haplotypes and five htSNPs in terms of the cell-surface expression of NKG2D protein on NK and CD8 T cells of peripheral blood among 732 atomic-bomb survivors. NKG2D expression on NK cells showed significant increases, in the order of LNK1/LNK1, LNK1/HNK1 and HNK1/HNK1 haplotypes (p for trend=0.003), or with major homozygous, heterozygous, and minor homozygous genotypes for individual htSNPs (p for trend=0.02-0.003). The same trend was observed for NKG2D expression on CD8 T cells. Our findings indicate that the NKG2D haplotypes are associated with the expression levels of NKG2D protein on NK and CD8 T cells, resulting in inter-individual variations in human cytotoxic response.

  6. Effects of NKG2D haplotypes on the cell-surface expression of NKG2D protein on natural killer and CD8 T cells of peripheral blood among atomic-bomb survivors.

    PubMed

    Imai, Kazue; Hayashi, Tomonori; Yamaoka, Mika; Kajimura, Junko; Yoshida, Kengo; Kusunoki, Yoichiro; Nakachi, Kei

    2012-06-01

    NKG2D is a primary activating receptor that triggers cell-mediated cytotoxicity in NK cells against tumor and virus-infected cells. We previously identified the NKG2D haplotypes in the natural killer gene complex region on chromosome 12p. Two major haplotype alleles, LNK1 and HNK1, were closely related to low and high natural cytotoxic activity phenotypes, respectively. Furthermore, the haplotype of HNK1/HNK1 has revealed a decreased risk of cancer compared with LNK1/LNK1. In the present study, using flow cytometry, we evaluated the functional effects of NKG2D haplotypes and five htSNPs in terms of the cell-surface expression of NKG2D protein on NK and CD8 T cells of peripheral blood among 732 atomic-bomb survivors. NKG2D expression on NK cells showed significant increases, in the order of LNK1/LNK1, LNK1/HNK1 and HNK1/HNK1 haplotypes (p for trend=0.003), or with major homozygous, heterozygous, and minor homozygous genotypes for individual htSNPs (p for trend=0.02-0.003). The same trend was observed for NKG2D expression on CD8 T cells. Our findings indicate that the NKG2D haplotypes are associated with the expression levels of NKG2D protein on NK and CD8 T cells, resulting in inter-individual variations in human cytotoxic response. PMID:22507622

  7. A Cartesian grid finite-difference method for 2D incompressible viscous flows in irregular geometries

    NASA Astrophysics Data System (ADS)

    Sanmiguel-Rojas, Enrique; Ortega-Casanova, Joaquin; del Pino, Carlos; Fernandez-Feria, Ramon

    2004-11-01

    A method for generating a non-uniform cartesian grid for irregular two-dimensional (2D) geometries such that all the boundary points are regular mesh points is given. The resulting non-uniform grid is used to discretize the Navier-Stokes equations for 2D incompressible viscous flows using finite difference approximations. To that end, finite-difference approximations of the derivatives on a non-uniform mesh are given. We test the method with two different examples: the shallow water flow on a lake with irregular contour, and the pressure driven flow through an irregular array of circular cylinders.

  8. Fluctuating Pressure Data from 2-D Nozzle Cold Flow Tests (Dual Bell)

    NASA Technical Reports Server (NTRS)

    Nesman, Tomas E.

    2001-01-01

    Rocket engines nozzle performance changes as a vehicle climbs through the atmosphere. An altitude compensating nozzle, ACN, is intended to improve on a fixed geometry bell nozzle that performs at optimum at only one trajectory point. In addition to nozzle performance, nozzle transient loads are an important consideration. Any nozzle experiences large transient toads when shocks pass through the nozzle at start and shutdown. Additional transient toads will occur at transitional flow conditions. The objectives of cold flow nozzle testing at MSFC are CFD benchmark / calibration and Unsteady flow / sideloads. Initial testing performed with 2-D inserts to 14" transonic wind tunnel. Recent review of 2-D data in preparation for nozzle test facility 3-D testing. This presentation shows fluctuating pressure data and some observations from 2-D dual-bell nozzle cold flow tests.

  9. A droplet-to-digital (D2D) microfluidic device for single cell assays.

    PubMed

    Shih, Steve C C; Gach, Philip C; Sustarich, Jess; Simmons, Blake A; Adams, Paul D; Singh, Seema; Singh, Anup K

    2015-01-01

    We have developed a new hybrid droplet-to-digital microfluidic platform (D2D) that integrates droplet-in-channel microfluidics with digital microfluidics (DMF) for performing multi-step assays. This D2D platform combines the strengths of the two formats-droplets-in-channel for facile generation of droplets containing single cells, and DMF for on-demand manipulation of droplets including control of different droplet volumes (pL-μL), creation of a dilution series of ionic liquid (IL), and parallel single cell culturing and analysis for IL toxicity screening. This D2D device also allows for automated analysis that includes a feedback-controlled system for merging and splitting of droplets to add reagents, an integrated Peltier element for parallel cell culture at optimum temperature, and an impedance sensing mechanism to control the flow rate for droplet generation and preventing droplet evaporation. Droplet-in-channel is well-suited for encapsulation of single cells as it allows the careful manipulation of flow rates of aqueous phase containing cells and oil to optimize encapsulation. Once single cell containing droplets are generated, they are transferred to a DMF chip via a capillary where they are merged with droplets containing IL and cultured at 30 °C. The DMF chip, in addition to permitting cell culture and reagent (ionic liquid/salt) addition, also allows recovery of individual droplets for off-chip analysis such as further culturing and measurement of ethanol production. The D2D chip was used to evaluate the effect of IL/salt type (four types: NaOAc, NaCl, [C2mim] [OAc], [C2mim] [Cl]) and concentration (four concentrations: 0, 37.5, 75, 150 mM) on the growth kinetics and ethanol production of yeast and as expected, increasing IL concentration led to lower biomass and ethanol production. Specifically, [C2mim] [OAc] had inhibitory effects on yeast growth at concentrations 75 and 150 mM and significantly reduced their ethanol production compared to cells grown

  10. 2D light scattering static cytometry for label-free single cell analysis with submicron resolution.

    PubMed

    Xie, Linyan; Yang, Yan; Sun, Xuming; Qiao, Xu; Liu, Qiao; Song, Kun; Kong, Beihua; Su, Xuantao

    2015-11-01

    Conventional optical cytometric techniques usually measure fluorescence or scattering signals at fixed angles from flowing cells in a liquid stream. Here we develop a novel cytometer that employs a scanning optical fiber to illuminate single static cells on a glass slide, which requires neither microfluidic fabrication nor flow control. This static cytometric technique measures two dimensional (2D) light scattering patterns via a small numerical aperture (0.25) microscope objective for label-free single cell analysis. Good agreement is obtained between the yeast cell experimental and Mie theory simulated patterns. It is demonstrated that the static cytometer with a microscope objective of a low resolution around 1.30 μm has the potential to perform high resolution analysis on yeast cells with distributed sizes. The capability of the static cytometer for size determination with submicron resolution is validated via measurements on standard microspheres with mean diameters of 3.87 and 4.19 μm. Our 2D light scattering static cytometric technique may provide an easy-to-use, label-free, and flow-free method for single cell diagnostics.

  11. CAS2D- NONROTATING BLADE-TO-BLADE, STEADY, POTENTIAL TRANSONIC CASCADE FLOW ANALYSIS CODE

    NASA Technical Reports Server (NTRS)

    Dulikravich, D. S.

    1994-01-01

    An exact, full-potential-equation model for the steady, irrotational, homoentropic, and homoenergetic flow of a compressible, inviscid fluid through a two-dimensional planar cascade together with its appropriate boundary conditions has been derived. The CAS2D computer program numerically solves an artificially time-dependent form of the actual full-potential-equation, providing a nonrotating blade-to-blade, steady, potential transonic cascade flow analysis code. Comparisons of results with test data and theoretical solutions indicate very good agreement. In CAS2D, the governing equation is discretized by using type-dependent, rotated finite differencing and the finite area technique. The flow field is discretized by providing a boundary-fitted, nonuniform computational mesh. This mesh is generated by using a sequence of conformal mapping, nonorthogonal coordinate stretching, and local, isoparametric, bilinear mapping functions. The discretized form of the full-potential equation is solved iteratively by using successive line over relaxation. Possible isentropic shocks are captured by the explicit addition of an artificial viscosity in a conservative form. In addition, a four-level, consecutive, mesh refinement feature makes CAS2D a reliable and fast algorithm for the analysis of transonic, two-dimensional cascade flows. The results from CAS2D are not directly applicable to three-dimensional, potential, rotating flows through a cascade of blades because CAS2D does not consider the effects of the Coriolis force that would be present in the three-dimensional case. This program is written in FORTRAN IV for batch execution and has been implemented on an IBM 370 series computer with a central memory requirement of approximately 200K of 8 bit bytes. The CAS2D program was developed in 1980.

  12. Extension and application of the Preissmann slot model to 2D transient mixed flows

    NASA Astrophysics Data System (ADS)

    Maranzoni, Andrea; Dazzi, Susanna; Aureli, Francesca; Mignosa, Paolo

    2015-08-01

    This paper presents an extension of the Preissmann slot concept for the modeling of highly transient two-dimensional (2D) mixed flows. The classic conservative formulation of the 2D shallow water equations for free surface flows is adapted by assuming that two fictitious vertical slots, aligned along the two Cartesian plane directions and normally intersecting, are added on the ceiling of each integration element. Accordingly, transitions between free surface and pressurized flow can be handled in a natural and straightforward way by using the same set of governing equations. The opportunity of coupling free surface and pressurized flows is actually useful not only in one-dimensional (1D) problems concerning sewer systems but also for modeling 2D flooding phenomena in which the pressurization of bridges, culverts, or other crossing hydraulic structures can be expected. Numerical simulations are performed by using a shock-capturing MUSCL-Hancock finite volume scheme combined with the FORCE (First-Order Centred) solver for the evaluation of the numerical fluxes. The validation of the mathematical model is accomplished on the basis of both exact solutions of 1D discontinuous initial value problems and reference radial solutions of idealized test cases with cylindrical symmetry. Furthermore, the capability of the model to deal with practical field-scale applications is assessed by simulating the transit of a bore under an arch bridge. Numerical results show that the proposed model is suitable for the prediction of highly transient 2D mixed flows.

  13. The Sobolev Stability Threshold for 2D Shear Flows Near Couette

    NASA Astrophysics Data System (ADS)

    Bedrossian, Jacob; Vicol, Vlad; Wang, Fei

    2016-08-01

    We consider the 2D Navier-Stokes equation on T × R , with initial datum that is ɛ -close in H^N to a shear flow (U(y), 0), where Vert U(y) - yVert _{H^{N+4}} ≪ 1 and N>1 . We prove that if ɛ ≪ ν ^{1/2} , where ν denotes the inverse Reynolds number, then the solution of the Navier-Stokes equation remains ɛ -close in H^1 to (e^{t ν partial _{yy}}U(y),0) for all t>0 . Moreover, the solution converges to a decaying shear flow for times t ≫ ν ^{-1/3} by a mixing-enhanced dissipation effect, and experiences a transient growth of gradients. In particular, this shows that the stability threshold in finite regularity scales no worse than ν ^{1/2} for 2D shear flows close to the Couette flow.

  14. Evaluation of 2D shallow-water model for spillway flow with a complex geometry

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Although the two-dimensional (2D) shallow water model is formulated based on several assumptions such as hydrostatic pressure distribution and vertical velocity is negligible, as a simple alternative to the complex 3D model, it has been used to compute water flows in which these assumptions may be ...

  15. Parallelized CCHE2D flow model with CUDA Fortran on Graphics Process Units

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This paper presents the CCHE2D implicit flow model parallelized using CUDA Fortran programming technique on Graphics Processing Units (GPUs). A parallelized implicit Alternating Direction Implicit (ADI) solver using Parallel Cyclic Reduction (PCR) algorithm on GPU is developed and tested. This solve...

  16. Use of finite volume radiation for predicting the Knudsen minimum in 2D channel flow

    SciTech Connect

    Malhotra, Chetan P.; Mahajan, Roop L.

    2014-12-09

    In an earlier paper we employed an analogy between surface-to-surface radiation and free-molecular flow to model Knudsen flow through tubes and onto planes. In the current paper we extend the analogy between thermal radiation and molecular flow to model the flow of a gas in a 2D channel across all regimes of rarefaction. To accomplish this, we break down the problem of gaseous flow into three sub-problems (self-diffusion, mass-motion and generation of pressure gradient) and use the finite volume method for modeling radiation through participating media to model the transport in each sub-problem as a radiation problem. We first model molecular self-diffusion in the stationary gas by modeling the transport of the molecular number density through the gas starting from the analytical asymptote for free-molecular flow to the kinetic theory limit of gaseous self-diffusion. We then model the transport of momentum through the gas at unit pressure gradient to predict Poiseuille flow and slip flow in the 2D gas. Lastly, we predict the generation of pressure gradient within the gas due to molecular collisions by modeling the transport of the forces generated due to collisions per unit volume of gas. We then proceed to combine the three radiation problems to predict flow of the gas over the entire Knudsen number regime from free-molecular to transition to continuum flow and successfully capture the Knudsen minimum at Kn ∼ 1.

  17. Simulation of the flow and mass transfer for KDP crystals undergoing 2D translation during growth

    NASA Astrophysics Data System (ADS)

    Zhou, Chuan; Li, Mingwei; Hu, Zhitao; Yin, Huawei; Wang, Bangguo; Cui, Qidong

    2016-09-01

    In this study, a novel motion mode for crystals during growth, i.e., 2D translation, is proposed. Numerical simulations of flow and mass transfer are conducted for the growth of large-scale potassium dihydrogen phosphate (KDP) crystals subjected to the new motion mode. Surface supersaturation and shear stress are obtained as functions of the translational velocity, distance, size, orientation of crystals. The dependence of these two parameters on the flow fields around the crystals is also discussed. The thicknesses of the solute boundary layer varied with translational velocity are described. The characteristics of solution flow and surface supersaturation distribution are summarized, where it suggests that the morphological stability of a crystal surface can be enhanced if the proposed 2D translation is applied to crystal growth.

  18. A depth-averaged 2-D model of flow and sediment transport in coastal waters

    NASA Astrophysics Data System (ADS)

    Sanchez, Alejandro; Wu, Weiming; Beck, Tanya M.

    2016-11-01

    A depth-averaged 2-D model has been developed to simulate unsteady flow and nonuniform sediment transport in coastal waters. The current motion is computed by solving the phase-averaged 2-D shallow water flow equations reformulated in terms of total-flux velocity, accounting for the effects of wave radiation stresses and general diffusion or mixing induced by current, waves, and wave breaking. The cross-shore boundary conditions are specified by assuming fully developed longshore current and wave setup that are determined using the reduced 1-D momentum equations. A 2-D wave spectral transformation model is used to calculate the wave height, period, direction, and radiation stresses, and a surface wave roller model is adopted to consider the effects of surface roller on the nearshore currents. The nonequilibrium transport of nonuniform total-load sediment is simulated, considering sediment entrainment by current and waves, the lag of sediment transport relative to the flow, and the hiding and exposure effect of nonuniform bed material. The flow and sediment transport equations are solved using an implicit finite volume method on a variety of meshes including nonuniform rectangular, telescoping (quadtree) rectangular, and hybrid triangular/quadrilateral meshes. The flow and wave models are integrated through a carefully designed steering process. The model has been tested in three field cases, showing generally good performance.

  19. F2D users manual: A two-dimensional compressible gas flow code

    NASA Astrophysics Data System (ADS)

    Suo-Anttila, A.

    1993-08-01

    The F2D computer code is a general purpose, two-dimensional, fully compressible thermal-fluids code that models most of the phenomena found in situations of coupled fluid flow and heat transfer. The code solves momentum, continuity, gas-energy, and structure-energy equations using a predictor-corrector solution algorithm. The corrector step includes a Poisson pressure equation. The finite difference form of the equation is presented along with a description of input and output. Several example problems are included that demonstrate the applicability of the code in problems ranging from free fluid flow, shock tubes, and flow in heated porous media.

  20. F2D. A Two-Dimensional Compressible Gas Flow Code

    SciTech Connect

    Suo-Anttila, A.

    1993-08-01

    F2D is a general purpose, two dimensional, fully compressible thermal-fluids code that models most of the phenomena found in situations of coupled fluid flow and heat transfer. The code solves momentum, continuity, gas-energy, and structure-energy equations using a predictor-correction solution algorithm. The corrector step includes a Poisson pressure equation. The finite difference form of the equation is presented along with a description of input and output. Several example problems are included that demonstrate the applicability of the code in problems ranging from free fluid flow, shock tubes and flow in heated porous media.

  1. F2D users manual: A two-dimensional compressible gas flow code

    SciTech Connect

    Suo-Anttila, A.

    1993-08-01

    The F2D computer code is a general purpose, two-dimensional, fully compressible thermal-fluids code that models most of the phenomena found in situations of coupled fluid flow and heat transfer. The code solves momentum, continuity, gas-energy, and structure-energy equations using a predictor-corrector solution algorithm. The corrector step includes a Poisson pressure equation. The finite difference form of the equation is presented along with a description of input and output. Several example problems are included that demonstrate the applicability of the code in problems ranging from free fluid flow, shock tubes and flow in heated porous media.

  2. Simulation of Pyroclastic Flows of Colima Volcano, Mexico, Using the TITAN2D Program

    NASA Astrophysics Data System (ADS)

    Rupp, B.; Bursik, M.; Patra, A.; Pitman, B.; Bauer, A.; Nichita, C.; Saucedo, R.; Macias, J.

    2003-04-01

    A new numerical code for simulating granular avalanches, TITAN2D, was used to model block-and-ash flows from the 1991-1999 eruptions of Colima Volcano, Mexico. The block-and-ash flows were simulated on a gridded Digital Elevation Model(DEM), which was prepared and imported using a standard GIS function library (GRASS). The TITAN2D program is based upon a model for an incompressible Coulomb continuum, a 'shallow-water' granular flow. The conservation equations for mass and momentum are solved with a Coulomb-type friction term at the interface between the granular material and the basal surface. It is assumed that conservation of energy can be neglected to first order because the coarse grain size typical of the basal avalanche results in minimal thermal effects on avalanche propagation. The resulting hyperbolic system of equations is solved using a parallel, adaptive mesh, Godunov scheme. The Message Passing Interface (MPI) API allows for computing on multiple processors, which increases computational power, decreases computing time, and allows the use of large data sets. Adaptive gridding allows for the concentration of computing power on regions of special interest. Mesh refinement captures the leading edge of the avalanche, as well as locations where the topography changes rapidly. Mesh unrefinement is applied where solution values are relatively constant or small. There were thousands of rockfalls and numerous block-and-ash flows during the 1991-1999 eruptions of Colima Volcano, with volumes ranging from a few cubic meters to 10^6 m^3. We have records of numerous flows, which include volume, run out distance, deposit area, and in some cases a videotape record of flow propagation. The flows originated from a vent plugging dome, lava flows or minor column collapse. All flows followed cross-slope concavities on the upper edifice, and channels or relative topographic lows on the lower edifice. The flows propagated for distances up to 4 km from the source. We are

  3. Investigation of the effect of wall friction on the flow rate in 2D and 3D Granular Flow

    NASA Astrophysics Data System (ADS)

    Carballo-Ramirez, Brenda; Pleau, Mollie; Easwar, Nalini; Birwa, Sumit; Shah, Neil; Tewari, Shubha

    We have measured the mass flow rate of spherical steel spheres under gravity in vertical, straight-walled 2 and 3-dimensional hoppers, where the flow velocity is controlled by the opening size. Our measurements focus on the role of friction and its placement along the walls of the hopper. In the 2D case, an increase in the coefficient of static friction from μ = 0.2 to 0.6 is seen to decrease the flow rate significantly. We have changed the placement of frictional boundaries/regions from the front and back walls of the 2D hopper to the side walls and floor to investigate the relative importance of the different regions in determining the flow rate. Fits to the Beverloo equation show significant departure from the expected exponent of 1.5 in the case of 2D flow. In contrast, 3D flow rates do not show much dependence on wall friction and its placement. We compare the experimental data to numerical simulations of gravity driven hopper granular flow with varying frictional walls constructed using LAMMPS*. *http://lammps.sandia.gov Supported by NSF MRSEC DMR 0820506.

  4. Moment Invariants for 2D Flow Fields via Normalization in Detail.

    PubMed

    Bujack, Roxana; Hotz, Ingrid; Scheuermann, Gerik; Hitzer, Eckhard

    2015-08-01

    The analysis of 2D flow data is often guided by the search for characteristic structures with semantic meaning. One way to approach this question is to identify structures of interest by a human observer, with the goal of finding similar structures in the same or other datasets. The major challenges related to this task are to specify the notion of similarity and define respective pattern descriptors. While the descriptors should be invariant to certain transformations, such as rotation and scaling, they should provide a similarity measure with respect to other transformations, such as deformations. In this paper, we propose to use moment invariants as pattern descriptors for flow fields. Moment invariants are one of the most popular techniques for the description of objects in the field of image recognition. They have recently also been applied to identify 2D vector patterns limited to the directional properties of flow fields. Moreover, we discuss which transformations should be considered for the application to flow analysis. In contrast to previous work, we follow the intuitive approach of moment normalization, which results in a complete and independent set of translation, rotation, and scaling invariant flow field descriptors. They also allow to distinguish flow features with different velocity profiles. We apply the moment invariants in a pattern recognition algorithm to a real world dataset and show that the theoretical results can be extended to discrete functions in a robust way. PMID:26357255

  5. Mouse tumor vasculature expresses NKG2D ligands and can be targeted by chimeric NKG2D-modified T cells.

    PubMed

    Zhang, Tong; Sentman, Charles L

    2013-03-01

    Tumor angiogenesis plays an important role in the development of solid tumors, and targeting the tumor vasculature has emerged as a strategy to prevent growth and progression of solid tumors. In this study, we show that murine tumor vasculature expresses Rae1, a ligand for a stimulatory NK receptor NKG2D. By genetic modification of T cells with an NKG2D-based chimeric Ag receptor, referred to as chNKG2D in which the NKG2D receptor is fused to the signaling domain of CD3ζ-chain, T cells were capable of targeting tumor vasculature leading to reduced tumor angiogenesis and tumor growth. This occurred even in tumors where the tumor cells themselves did not express NKG2D ligands. H5V, an endothelial cell line, expresses Rae1 and was lysed by chNKG2D-bearing T cells in a perforin-dependent manner. In vitro capillary tube formation was inhibited by chNKG2D T cells through IFN-γ and cell-cell contact mechanisms. The in vivo antiangiogenesis effects mediated by chNKG2D-bearing T cells at the tumor site were dependent on IFN-γ and perforin. These results provide a novel mechanism for NKG2D-based targeting of solid tumors.

  6. Grid Cell Responses in 1D Environments Assessed as Slices through a 2D Lattice.

    PubMed

    Yoon, KiJung; Lewallen, Sam; Kinkhabwala, Amina A; Tank, David W; Fiete, Ila R

    2016-03-01

    Grid cells, defined by their striking periodic spatial responses in open 2D arenas, appear to respond differently on 1D tracks: the multiple response fields are not periodically arranged, peak amplitudes vary across fields, and the mean spacing between fields is larger than in 2D environments. We ask whether such 1D responses are consistent with the system's 2D dynamics. Combining analytical and numerical methods, we show that the 1D responses of grid cells with stable 1D fields are consistent with a linear slice through a 2D triangular lattice. Further, the 1D responses of comodular cells are well described by parallel slices, and the offsets in the starting points of the 1D slices can predict the measured 2D relative spatial phase between the cells. From these results, we conclude that the 2D dynamics of these cells is preserved in 1D, suggesting a common computation during both types of navigation behavior. PMID:26898777

  7. Flow Quantification from 2D Phase Contrast MRI in Renal Arteries Using Clustering

    NASA Astrophysics Data System (ADS)

    Zöllner, Frank G.; Monnsen, Jan Ankar; Lundervold, Arvid; Rørvik, Jarle

    We present an approach based on clustering to segment renal arteries from 2D PC Cine MR images to measure blood velocity and flow. Such information are important in grading renal artery stenosis and support the decision on surgical interventions like percutan transluminal angioplasty. Results show that the renal arteries could be extracted automatically and the corresponding velocity profiles could be calculated. Furthermore, the clustering could detect possible phase wrap effects automatically as well as differences in the blood flow patterns within the vessel.

  8. Creeping motion and deformation of liquid drops in flow through 2D model porous media

    SciTech Connect

    Fong, I. )

    1988-01-01

    The motion, deformation and breakup of immiscible drops suspended in low Reynolds number flow through cylinder arrays has been studied experimentally to assess the applicability of the 2D model as a prototype for 2-phase flow through porous media. Both Newtonian and non-Newtonian fluid systems are considered. The relationship between key flow and geometric parameters and the critical condition for breakup, the resulting drop site distribution and the drop mobility is investigated. It is observed that the headon impact of a drop with a cylinder is an effective precursor to severe drop deformation and even breakup. The sequence of flow leading to impact is also important in determining the effectiveness of impact to result in breakup. When many drops fragments are present, the interaction between nearby drops strongly influences the final disposition of the fragments. Fluid elasticity appears to enhance the elongation of drops to form strands, but also to stabilize the strand against breakup.

  9. 1,25(OH)2D3 increases membrane associated protein kinase C in MDBK cells.

    PubMed

    Simboli-Campbell, M; Franks, D J; Welsh, J

    1992-01-01

    To determine whether 1,25-dihydroxycholecalciferol [1,25(OH)2D3] affects protein kinase C (PKC) activity in kidney, as has been demonstrated in HL-60 cells we measured 1,25(OH)2D3 binding, PKC activity and PKC immunoreactivity in Madin Darby bovine kidney (MDBK) cells, a normal renal epithelial cell line derived from bovine kidney. Our data demonstrate that MDBK cells exhibit specific high affinity binding for 1,25(OH)2D3, indicating the presence of the vitamin D receptor (VDR). Treatment of MDBK cells with 1,25(OH)2D3 for 24 h increased membrane PKC activity and immunoreactivity. The effect of 1,25(OH)2D3 was dose-dependent, with a peak effect observed at 10(-7)M 1,25(OH)2D3. The 1,25(OH)2D3 induced increase in membrane PKC was paralleled by a comparable decrease in cytosolic PKC activity and amount. Although time course studies were consistent with a VDR mediated effect of 1,25(OH)2D3 on PKC protein synthesis, total PKC activity was not increased by 1,25(OH)2D3, suggesting an effect on PKC translocation or localization. These results suggest that 1,25(OH)2D3 modulates PKC mediated events in kidney, a classic target for this steroid hormone.

  10. Upregulation of NKG2D ligands and enhanced natural killer cell cytotoxicity by hydralazine and valproate.

    PubMed

    Chávez-Blanco, A; De la Cruz-Hernández, E; Domínguez, G I; Rodríguez-Cortez, O; Alatorre, B; Pérez-Cárdenas, E; Chacón-Salinas, R; Trejo-Becerril, C; Taja-Chayeb, L; Trujillo, J E; Contreras-Paredes, A; Dueñas-González, A

    2011-12-01

    Natural killer cells play a role in the immune antitumor response by recognizing and eliminating tumor cells through the engagement of NKG2D receptors with their ligands on target cells. This work aimed to investigate whether epigenetic drugs are able to increase MICA and MICB expression as well as NK cell cytotoxicity. Prostate, colon, breast and cervical cancer cell lines were analyzed for the expression of MICA and MICB at the mRNA and protein levels by RT-PCR, Western blot, flow cytometry and ELISA. The activating mark H3K4m2 at the MICA and MICB promoters was investigated by ChIP assays. Cytotoxicity of NK cells against the target epithelial cancer cells was investigated with the CD107 cytotoxicity assay. The results show that hydralazine and valproic acid not only increase the expression of MICA and MICB ligands of target cells, but also reduce their shedding to the supernatant. This upregulation occurs at the transcriptional level as revealed by increase of the H3K4 activating mark at the promoter of MICA and MICB genes. These effects are paralleled by increased cytotoxicity of NK cells, which was attenuated at different degrees by using blocking antibodies against the NKG2D receptor and ligands. In conclusion, our results demonstrate the ability of hydralazine and valproate to increase the NK activity against epithelial cancer cell lines and suggest that these drugs could reduce the levels of soluble MICA and MICB helping in avoiding tumor-induced suppression of NK cytotoxicity against the tumor.

  11. Inhibitory effects of phytochemicals on metabolic capabilities of CYP2D6*1 and CYP2D6*10 using cell-based models in vitro

    PubMed Central

    Qu, Qiang; Qu, Jian; Han, Lu; Zhan, Min; Wu, Lan-xiang; Zhang, Yi-wen; Zhang, Wei; Zhou, Hong-hao

    2014-01-01

    Aim: Herbal products have been widely used, and the safety of herb-drug interactions has aroused intensive concerns. This study aimed to investigate the effects of phytochemicals on the catalytic activities of human CYP2D6*1 and CYP2D6*10 in vitro. Methods: HepG2 cells were stably transfected with CYP2D6*1 and CYP2D6*10 expression vectors. The metabolic kinetics of the enzymes was studied using HPLC and fluorimetry. Results: HepG2-CYP2D6*1 and HepG2-CYP2D6*10 cell lines were successfully constructed. Among the 63 phytochemicals screened, 6 compounds, including coptisine sulfate, bilobalide, schizandrin B, luteolin, schizandrin A and puerarin, at 100 μmol/L inhibited CYP2D6*1- and CYP2D6*10-mediated O-demethylation of a coumarin compound AMMC by more than 50%. Furthermore, the inhibition by these compounds was dose-dependent. Eadie-Hofstee plots demonstrated that these compounds competitively inhibited CYP2D6*1 and CYP2D6*10. However, their Ki values for CYP2D6*1 and CYP2D6*10 were very close, suggesting that genotype-dependent herb-drug inhibition was similar between the two variants. Conclusion: Six phytochemicals inhibit CYP2D6*1 and CYP2D6*10-mediated catalytic activities in a dose-dependent manner in vitro. Thus herbal products containing these phytochemicals may inhibit the in vivo metabolism of co-administered drugs whose primary route of elimination is CYP2D6. PMID:24786236

  12. Plane-wave transverse oscillation for high-frame-rate 2-D vector flow imaging.

    PubMed

    Lenge, Matteo; Ramalli, Alessandro; Tortoli, Piero; Cachard, Christian; Liebgott, Hervé

    2015-12-01

    Transverse oscillation (TO) methods introduce oscillations in the pulse-echo field (PEF) along the direction transverse to the ultrasound propagation direction. This may be exploited to extend flow investigations toward multidimensional estimates. In this paper, the TOs are coupled with the transmission of plane waves (PWs) to reconstruct high-framerate RF images with bidirectional oscillations in the pulse-echo field. Such RF images are then processed by a 2-D phase-based displacement estimator to produce 2-D vector flow maps at thousands of frames per second. First, the capability of generating TOs after PW transmissions was thoroughly investigated by varying the lateral wavelength, the burst length, and the transmission frequency. Over the entire region of interest, the generated lateral wavelengths, compared with the designed ones, presented bias and standard deviation of -3.3 ± 5.7% and 10.6 ± 7.4% in simulations and experiments, respectively. The performance of the ultrafast vector flow mapping method was also assessed by evaluating the differences between the estimated velocities and the expected ones. Both simulations and experiments show overall biases lower than 20% when varying the beam-to-flow angle, the peak velocity, and the depth of interest. In vivo applications of the method on the common carotid and the brachial arteries are also presented. PMID:26670852

  13. Plane-wave transverse oscillation for high-frame-rate 2-D vector flow imaging.

    PubMed

    Lenge, Matteo; Ramalli, Alessandro; Tortoli, Piero; Cachard, Christian; Liebgott, Hervé

    2015-12-01

    Transverse oscillation (TO) methods introduce oscillations in the pulse-echo field (PEF) along the direction transverse to the ultrasound propagation direction. This may be exploited to extend flow investigations toward multidimensional estimates. In this paper, the TOs are coupled with the transmission of plane waves (PWs) to reconstruct high-framerate RF images with bidirectional oscillations in the pulse-echo field. Such RF images are then processed by a 2-D phase-based displacement estimator to produce 2-D vector flow maps at thousands of frames per second. First, the capability of generating TOs after PW transmissions was thoroughly investigated by varying the lateral wavelength, the burst length, and the transmission frequency. Over the entire region of interest, the generated lateral wavelengths, compared with the designed ones, presented bias and standard deviation of -3.3 ± 5.7% and 10.6 ± 7.4% in simulations and experiments, respectively. The performance of the ultrafast vector flow mapping method was also assessed by evaluating the differences between the estimated velocities and the expected ones. Both simulations and experiments show overall biases lower than 20% when varying the beam-to-flow angle, the peak velocity, and the depth of interest. In vivo applications of the method on the common carotid and the brachial arteries are also presented.

  14. TRENT2D WG: a smart web infrastructure for debris-flow modelling and hazard assessment

    NASA Astrophysics Data System (ADS)

    Zorzi, Nadia; Rosatti, Giorgio; Zugliani, Daniel; Rizzi, Alessandro; Piffer, Stefano

    2016-04-01

    Mountain regions are naturally exposed to geomorphic flows, which involve large amounts of sediments and induce significant morphological modifications. The physical complexity of this class of phenomena represents a challenging issue for modelling, leading to elaborate theoretical frameworks and sophisticated numerical techniques. In general, geomorphic-flows models proved to be valid tools in hazard assessment and management. However, model complexity seems to represent one of the main obstacles to the diffusion of advanced modelling tools between practitioners and stakeholders, although the UE Flood Directive (2007/60/EC) requires risk management and assessment to be based on "best practices and best available technologies". Furthermore, several cutting-edge models are not particularly user-friendly and multiple stand-alone software are needed to pre- and post-process modelling data. For all these reasons, users often resort to quicker and rougher approaches, leading possibly to unreliable results. Therefore, some effort seems to be necessary to overcome these drawbacks, with the purpose of supporting and encouraging a widespread diffusion of the most reliable, although sophisticated, modelling tools. With this aim, this work presents TRENT2D WG, a new smart modelling solution for the state-of-the-art model TRENT2D (Armanini et al., 2009, Rosatti and Begnudelli, 2013), which simulates debris flows and hyperconcentrated flows adopting a two-phase description over a mobile bed. TRENT2D WG is a web infrastructure joining advantages offered by the software-delivering model SaaS (Software as a Service) and by WebGIS technology and hosting a complete and user-friendly working environment for modelling. In order to develop TRENT2D WG, the model TRENT2D was converted into a service and exposed on a cloud server, transferring computational burdens from the user hardware to a high-performing server and reducing computational time. Then, the system was equipped with an

  15. A fast and accurate method to predict 2D and 3D aerodynamic boundary layer flows

    NASA Astrophysics Data System (ADS)

    Bijleveld, H. A.; Veldman, A. E. P.

    2014-12-01

    A quasi-simultaneous interaction method is applied to predict 2D and 3D aerodynamic flows. This method is suitable for offshore wind turbine design software as it is a very accurate and computationally reasonably cheap method. This study shows the results for a NACA 0012 airfoil. The two applied solvers converge to the experimental values when the grid is refined. We also show that in separation the eigenvalues remain positive thus avoiding the Goldstein singularity at separation. In 3D we show a flow over a dent in which separation occurs. A rotating flat plat is used to show the applicability of the method for rotating flows. The shown capabilities of the method indicate that the quasi-simultaneous interaction method is suitable for design methods for offshore wind turbine blades.

  16. Field Evaluation of a Novel 2D Preferential Flow Snowpack Hydrology Model

    NASA Astrophysics Data System (ADS)

    Leroux, N.; Pomeroy, J. W.; Kinar, N. J.

    2015-12-01

    Accurate estimation of snowmelt flux is of primary importance for runoff hydrograph prediction, which is used for water management and flood forecasting. Lateral flows and preferential flow pathways in porous media flow have proven critical for improving soil and groundwater flow models, but though many physically-based layered snowmelt models have been developed, only 1D matrix flow is accounted for in these models. Therefore, there is a need for snowmelt models that include these processes so as to examine the potential to improve snowmelt hydrological modelling. A 2D model is proposed that enables an improved understanding of energy and water flows within deep heterogeneous snowpacks, including those on slopes. A dual pathway theory is presented that simulates the formation of preferential flow paths, vertical and lateral water flows through the snow matrix and flow fingers, internal energy fluxes, melt, wet snow metamorphism, and internal refreezing. The dual pathway model utilizes an explicit finite volume method to solve for the energy and water flux equations over a non-orthogonal grid. It was run and evaluated using in-situ data collected from snowpit - accessed gravimetric, thermometric, photographic, and dielectric observations and novel non-invasive acoustic observations of layering, temperature, flowpath geometry, density and wetness at the Fortress Mountain Snow Laboratory, Alberta, Canada. The melt of a natural snowpack was artificially generated after detailed observation of snowpack initial conditions such as snow layer properties, temperature, and liquid water content. Snowpack ablation and liquid water content distribution over time were then measured and used for model parameterization and validation. Energy available at the snow surface and soil slope angle were set as mondel inputs. Model verification was based on snowpack property evolution. The heterogeneous flow model can be an important tool to help understand snowmelt flow processes, how

  17. Aspects of Turbulent Flow over 2D and 3D Bedforms

    NASA Astrophysics Data System (ADS)

    Venditti, J. G.; Church, M. A.

    2004-05-01

    Sediment transport in sand bedded alluvial channels is strongly conditioned by bedforms. Understanding the turbulent flow field over bedforms is crucial to understanding flow resistance in rivers. Most of the research on flow over bedforms has focused on straight crested, two-dimensional bedforms, and the characteristics of the turbulent flow field are fairly well understood. In contrast, few studies have examined flow over 3D bedforms, which typically have irregular heights, lengths, and crestlines. This paper reports on experiments undertaken to examine how 3D dune morphology affects the turbulent flow field and, ultimately, flow resistance. An experiment was designed to examine flow over fixed bedforms 0.45 m long and 25 mm high in a 0.5 m wide and 17 m long flume. In each experimental run, discharge and dune size were held constant, but the crest shape was varied. Flow over six bedform crest shapes was examined, including a 2D crest, a saddle shaped crest, a lobe shaped crest, a regular 3D crest alignment, an irregular 3D crest alignment and a sinuous crest. Measurements of velocity were made at a sampling rate of 50 Hz using an acoustic Doppler velocimeter at 350-500 points over a dune in each morphology. Three-dimensional bedforms significantly modify the flow field over a dune. Lobe shaped configurations cause lateral and vertical divergence of momentum and turbulent energy, thereby enhancing the level of turbulence compared to a 2D bedform. Saddle shaped crestlines cause lateral and vertical convergence of momentum and turbulent energy towards a small area in the lee, thereby reducing the level of turbulence. Other bedform morphologies (regular, irregular and sinuous crests) exhibited characteristics of both lobes and saddles, but the net effect was to reduce levels of turbulence. Total drag, calculated from spatially averaged Reynolds stress profiles, can be enhanced or reduced by as much as 50 %. These results suggest that current conceptions of bedforms

  18. CAS2D: FORTRAN program for nonrotating blade-to-blade, steady, potential transonic cascade flows

    NASA Technical Reports Server (NTRS)

    Dulikravich, D. S.

    1980-01-01

    An exact, full-potential-equation (FPE) model for the steady, irrotational, homentropic and homoenergetic flow of a compressible, homocompositional, inviscid fluid through two dimensional planar cascades of airfoils was derived, together with its appropriate boundary conditions. A computer program, CAS2D, was developed that numerically solves an artificially time-dependent form of the actual FPE. The governing equation was discretized by using type-dependent, rotated finite differencing and the finite area technique. The flow field was discretized by providing a boundary-fitted, nonuniform computational mesh. The mesh was generated by using a sequence of conforming mapping, nonorthogonal coordinate stretching, and local, isoparametric, bilinear mapping functions. The discretized form of the FPE was solved iteratively by using successive line overrelaxation. The possible isentropic shocks were correctly captured by adding explicitly an artificial viscosity in a conservative form. In addition, a three-level consecutive, mesh refinement feature makes CAS2D a reliable and fast algorithm for the analysis of transonic, two dimensional cascade flows.

  19. Line relaxation methods for the solution of 2D and 3D compressible flows

    NASA Technical Reports Server (NTRS)

    Hassan, O.; Probert, E. J.; Morgan, K.; Peraire, J.

    1993-01-01

    An implicit finite element based algorithm for the compressible Navier-Stokes equations is outlined, and the solution of the resulting equation by a line relaxation on general meshes of triangles or tetrahedra is described. The problem of generating and adapting unstructured meshes for viscous flows is reexamined, and an approach for both 2D and 3D simulations is proposed. An efficient approach appears to be the use of an implicit/explicit procedure, with the implicit treatment being restricted to those regions of the mesh where viscous effects are known to be dominant. Numerical examples demonstrating the computational performance of the proposed techniques are given.

  20. Dephosphorylation of MAP2D enhances its binding to vimentin in preovulatory ovarian granulosa cells.

    PubMed

    Flynn, Maxfield P; Fiedler, Sarah E; Karlsson, Amelia B; Carr, Daniel W; Maizels, Evelyn T; Hunzicker-Dunn, Mary

    2016-08-01

    Preovulatory granulosa cells express the low-molecular-mass MAP2D variant of microtubule-associated protein 2 (MAP2). Activation of the luteinizing hormone choriogonadotropin receptor by human choriogonadotropin (hCG) promotes dephosphorylation of MAP2D on Thr256 and Thr259. We sought to evaluate the association of MAP2D with the cytoskeleton, and the effect of hCG on this association. MAP2D partially colocalized, as assessed by confocal immunofluorescence microscopy, with the vimentin intermediate filament and microtubule cytoskeletons in naive cells. In vitro binding studies showed that MAP2D bound directly to vimentin and β-tubulin. Phosphorylation of recombinant MAP2D on Thr256 and Thr259, which mimics the phosphorylation status of MAP2D in naive cells, reduces binding of MAP2D to vimentin and tubulin by two- and three-fold, respectively. PKA-dependent phosphorylation of vimentin (Ser32 and Ser38) promoted binding of vimentin to MAP2D and increased contraction of granulosa cells with reorganization of vimentin filaments and MAP2D from the periphery into a thickened layer surrounding the nucleus and into prominent cellular extensions. Chemical disruption of vimentin filament organization increased progesterone production. Taken together, these results suggest that hCG-stimulated dephosphorylation of MAP2D at Thr256 and Thr259, phosphorylation of vimentin at Ser38 and Ser72, and the resulting enhanced binding of MAP2D to vimentin might contribute to the progesterone synthetic response required for ovulation.

  1. Disruption of KMT2D perturbs germinal center B cell development and promotes lymphomagenesis.

    PubMed

    Zhang, Jiyuan; Dominguez-Sola, David; Hussein, Shafinaz; Lee, Ji-Eun; Holmes, Antony B; Bansal, Mukesh; Vlasevska, Sofija; Mo, Tongwei; Tang, Hongyan; Basso, Katia; Ge, Kai; Dalla-Favera, Riccardo; Pasqualucci, Laura

    2015-10-01

    Mutations in the gene encoding the KMT2D (or MLL2) methyltransferase are highly recurrent and occur early during tumorigenesis in diffuse large B cell lymphoma (DLBCL) and follicular lymphoma (FL). However, the functional consequences of these mutations and their role in lymphomagenesis are unknown. Here we show that FL- and DLBCL-associated KMT2D mutations impair KMT2D enzymatic activity, leading to diminished global H3K4 methylation in germinal-center (GC) B cells and DLBCL cells. Conditional deletion of Kmt2d early during B cell development, but not after initiation of the GC reaction, results in an increase in GC B cells and enhances B cell proliferation in mice. Moreover, genetic ablation of Kmt2d in mice overexpressing Bcl2 increases the incidence of GC-derived lymphomas resembling human tumors. These findings suggest that KMT2D acts as a tumor suppressor gene whose early loss facilitates lymphomagenesis by remodeling the epigenetic landscape of the cancer precursor cells. Eradication of KMT2D-deficient cells may thus represent a rational therapeutic approach for targeting early tumorigenic events. PMID:26366712

  2. Reconfiguration of a flexible fiber immersed in a 2D dense granular flow close to the jamming transition

    NASA Astrophysics Data System (ADS)

    Kolb, Evelyne; Algarra, Nicolas; Vandembroucq, Damien; Lazarus, Arnaud

    2015-11-01

    We propose a new fluid/structure interaction in the unusual case of a dense granular medium flowing against an elastic fibre acting as a flexible intruder. We experimentally studied the deflection of a mylar flexible beam clamped at one side, the other free side facing a 2D granular flow in a horizontal cell moving at a constant velocity. We investigated the reconfiguration of the fibre as a function of the fibre's rigidity and of the granular packing fraction close but below the jamming in 2D. Imposing the fibre geometry like its length or thickness sets the critical buckling force the fibre is able to resist if it was not supported by lateral grains, while increasing the granular packing fraction might laterally consolidate the fibre and prevent it from buckling. But on the other side, the approach to jamming transition by increasing the granular packing fraction will be characterized by a dramatically increasing size of the cluster of connected grains forming a solid block acting against the fibre, which might promote the fibre's deflection. Thus, we investigated the granular flow fields, the fibre's deflexion as well as the forces experienced by the fibre and compared them with theoretical predictions from elastica for different loadings along the fibre. PMMH, CNRS UMR 7636, UPMC, ESPCI-ParisTech, 10 rue Vauquelin, 75231 Paris Cedex 05, France.

  3. Chaotic advection and heat transfer in two similar 2-D periodic flows and in their corresponding 3-D periodic flows

    NASA Astrophysics Data System (ADS)

    Vinsard, G.; Dufour, S.; Saatdjian, E.; Mota, J. P. B.

    2016-03-01

    Chaotic advection can effectively enhance the heat transfer rate between a boundary and fluids with high Prandtl number. These fluids are usually highly viscous and thus turbulent agitation is not a viable solution since the energy required to mix the fluid would be prohibitive. Here, we analyze previously obtained results on chaotic advection and heat transfer in two similar 2-D periodic flows and on their corresponding 3-D periodic flows when an axial velocity component is superposed. The two flows studied are the flow between eccentric rotating cylinders and the flow between confocal ellipses. For both of these flows the analysis is simplified because the Stokes equations can be solved analytically to obtain a closed form solution. For both 2-D periodic flows, we show that chaotic heat transfer is enhanced by the displacement of the saddle point location during one period. Furthermore, the enhancement by chaotic advection in the elliptical geometry is approximately double that obtained in the cylindrical geometry because there are two saddle points instead of one. We also explain why, for high eccentricity ratios, there is no heat transfer enhancement in the cylindrical geometry. When an axial velocity component is added to both of these flows so that they become 3-D, previous work has shown that there is an optimum modulation frequency for which chaotic advection and heat transfer enhancement is a maximum. Here we show that the optimum modulation frequency can be derived from results without an axial flow. We also explain by physical arguments other previously unanswered questions in the published data.

  4. A shed NKG2D ligand that promotes natural killer cell activation and tumor rejection

    PubMed Central

    Deng, Weiwen; Gowen, Benjamin G.; Zhang, Li; Wang, Lin; Lau, Stephanie; Iannello, Alexandre; Xu, Jianfeng; Rovis, Tihana L.; Xiong, Na; Raulet, David H.

    2016-01-01

    Immune cells, including natural killer (NK) cells, recognize transformed cells and eliminate them in a process termed immunosurveillance. It is thought that tumor cells evade immunosurveillance by shedding membrane ligands that bind to the NKG2D activating receptor on NK cells and/or T cells, and desensitize these cells. In contrast, we show that in mice, shedding of MULT1, a high affinity NKG2D ligand, causes NK cell activation and tumor rejection. Recombinant soluble MULT1 stimulated tumor rejection in mice. Soluble MULT1 functions, at least in part, by competitively reversing a global desensitization of NK cells imposed by engagement of membrane NKG2D ligands on tumor-associated cells, such as myeloid cells. The results overturn conventional wisdom that soluble ligands are inhibitory, and suggest a new approach for cancer immunotherapy. PMID:25745066

  5. Evolution of vortices in 2D boundary layer and in the Couette flow

    NASA Astrophysics Data System (ADS)

    Zametaev, Vladimir B.; Gorbushin, Anton R.

    2016-10-01

    A 2D incompressible laminar boundary layer and the Couette flow having the low velocity fluctuations are considered using asymptotic methods at high Reynolds number. Two classes of solutions for the first order inviscid perturbations have been derived. The integral-differential equation with initial data describing evolution of vortices in time have been solved numerically. It was found that the discontinuities are formed in a smooth solution for a vertical velocity component with the time increase. This first type solution explains instability mechanism in the Couette flow. The second class of solutions contains a singularity at the boundary layer bottom which reminds a source-sink with a variable intensity. The singularity can absorb the fluid from the main part of the boundary layer and eject it back with a possibly "new" vorticity.

  6. Self-Organization in 2D Traffic Flow Model with Jam-Avoiding Drive

    NASA Astrophysics Data System (ADS)

    Nagatani, Takashi

    1995-04-01

    A stochastic cellular automaton (CA) model is presented to investigate the traffic jam by self-organization in the two-dimensional (2D) traffic flow. The CA model is the extended version of the 2D asymmetric exclusion model to take into account jam-avoiding drive. Each site contains either a car moving to the up, a car moving to the right, or is empty. A up car can shift right with probability p ja if it is blocked ahead by other cars. It is shown that the three phases (the low-density phase, the intermediate-density phase and the high-density phase) appear in the traffic flow. The intermediate-density phase is characterized by the right moving of up cars. The jamming transition to the high-density jamming phase occurs with higher density of cars than that without jam-avoiding drive. The jamming transition point p 2c increases with the shifting probability p ja. In the deterministic limit of p ja=1, it is found that a new jamming transition occurs from the low-density synchronized-shifting phase to the high-density moving phase with increasing density of cars. In the synchronized-shifting phase, all up cars do not move to the up but shift to the right by synchronizing with the move of right cars. We show that the jam-avoiding drive has an important effect on the dynamical jamming transition.

  7. Quasi-simultaneous interaction method for solving 2D boundary layer flows over plates and airfoils

    NASA Astrophysics Data System (ADS)

    Bijleveld, H. A.; Veldman, A. E. P.

    2012-11-01

    This paper studies unsteady 2D boundary layer flows over dented plates and a NACA 0012 airfoil. An inviscid flow is assumed to exist outside the boundary layer and is solved iteratively with the boundary layer flow together with the interaction method until a matching solution is achieved. Hereto a quasi-simultaneous interaction method is applied, in which the integral boundary layer equations are solved together with an interaction-law equation. The interaction-law equation is an approximation of the external flow and based on thin-airfoil theory. It is an algebraic relation between the velocity and displacement thickness. The interaction-law equation ensures that the eigenvalues of the system of equations do not have a sign change and that no singularities occur. Three numerical schemes are used to solve the boundary layer flow with the interaction method. These are: a standard scheme, a splitting method and a characteristics solver. All schemes use a finite difference discretization. The three schemes yield comparable results for the simulations carried out. The standard scheme is deviating most from the splitting and characteristics solvers. The results show that the eigenvalues remain positive, even in separation. As expected, the addition of the interaction-law equation prevents a sign change of the eigenvalues. The quasi-simultaneous interaction scheme is applicable to the three numerical schemes tested.

  8. Integrated Coupling of Surface and Subsurface Flow with HYDRUS-2D

    NASA Astrophysics Data System (ADS)

    Hartmann, Anne; Šimůnek, Jirka; Wöhling, Thomas; Schütze, Niels

    2016-04-01

    Describing interactions between surface and subsurface flow processes is important to adequately define water flow in natural systems. Since overland flow generation is highly influenced by rainfall and infiltration, both highly spatially heterogeneous processes, overland flow is unsteady and varies spatially. The prediction of overland flow needs to include an appropriate description of the interactions between the surface and subsurface flow. Coupling surface and subsurface water flow is a challenging task. Different approaches have been developed during the last few years, each having its own advantages and disadvantages. A new approach by Weill et al. (2009) to couple overland flow and subsurface flow based on a generalized Richards equation was implemented into the well-known subsurface flow model HYDRUS-2D (Šimůnek et al., 2011). This approach utilizes the one-dimensional diffusion wave equation to model overland flow. The diffusion wave model is integrated in HYDRUS-2D by replacing the terms of the Richards equation in a pre-defined runoff layer by terms defining the diffusion wave equation. Using this approach, pressure and flux continuity along the interface between both flow domains is provided. This direct coupling approach provides a strong coupling of both systems based on the definition of a single global system matrix to numerically solve the coupled flow problem. The advantage of the direct coupling approach, compared to the loosely coupled approach, is supposed to be a higher robustness, when many convergence problems can be avoided (Takizawa et al., 2014). The HYDRUS-2D implementation was verified using a) different test cases, including a direct comparison with the results of Weill et al. (2009), b) an analytical solution of the kinematic wave equation, and c) the results of a benchmark test of Maxwell et al. (2014), that included several known coupled surface subsurface flow models. Additionally, a sensitivity analysis evaluating the effects

  9. Differentiation of normal and leukemic cells by 2D light scattering label-free static cytometry.

    PubMed

    Xie, Linyan; Liu, Qiao; Shao, Changshun; Su, Xuantao

    2016-09-19

    Two-dimensional (2D) light scattering patterns of single microspheres, normal granulocytes and leukemic cells are obtained by label-free static cytometry. Statistical results of experimental 2D light scattering patterns obtained from standard microspheres with a mean diameter of 4.19 μm agree well with theoretical simulations. High accuracy rates (greater than 92%) for label-free differentiation of normal granulocytes and leukemic cells, both the acute and chronic leukemic cells, are achieved by analyzing the 2D light scattering patterns. Our label-free static cytometry is promising for leukemia screening in clinics. PMID:27661908

  10. Simulation of abrasive flow machining process for 2D and 3D mixture models

    NASA Astrophysics Data System (ADS)

    Dash, Rupalika; Maity, Kalipada

    2015-12-01

    Improvement of surface finish and material removal has been quite a challenge in a finishing operation such as abrasive flow machining (AFM). Factors that affect the surface finish and material removal are media viscosity, extrusion pressure, piston velocity, and particle size in abrasive flow machining process. Performing experiments for all the parameters and accurately obtaining an optimized parameter in a short time are difficult to accomplish because the operation requires a precise finish. Computational fluid dynamics (CFD) simulation was employed to accurately determine optimum parameters. In the current work, a 2D model was designed, and the flow analysis, force calculation, and material removal prediction were performed and compared with the available experimental data. Another 3D model for a swaging die finishing using AFM was simulated at different viscosities of the media to study the effects on the controlling parameters. A CFD simulation was performed by using commercially available ANSYS FLUENT. Two phases were considered for the flow analysis, and multiphase mixture model was taken into account. The fluid was considered to be a

  11. Debris Flow Hazard Map Simulation using FLO-2D For Selected Areas in the Philippines

    NASA Astrophysics Data System (ADS)

    Khallil Ferrer, Peter; Llanes, Francesca; dela Resma, Marvee; Realino, Victoriano, II; Obrique, Julius; Ortiz, Iris Jill; Aquino, Dakila; Narod Eco, Rodrigo; Mahar Francisco Lagmay, Alfredo

    2014-05-01

    On December 4, 2012, Super Typhoon Bopha wreaked havoc in the southern region of Mindanao, leaving 1,067 people dead and causing USD 800 million worth of damage. Classified as a Category 5 typhoon by the Joint Typhoon Warning Center (JTWC), Bopha brought intense rainfall and strong winds that triggered landslides and debris flows, particularly in Barangay (village) Andap, New Bataan municipality, in the southern Philippine province of Compostela Valley. The debris flow destroyed school buildings and covered courts and an evacuation center. Compostela Valley also suffered the most casualties of any province: 612 out of a total of 1,067. In light of the disaster in Compostela, measures were immediately devised to improve available geohazard maps to raise public awareness about landslides and debris flows. A debris flow is a very rapid to extremely rapid flow of saturated non-plastic debris in a steep channel. They are generated when heavy rainfall saturates sediments, causing them to flow down river channels within an alluvial fan situated at the base of the slope of a mountain drainage network. Many rural communities in the Philippines, such as Barangay Andap, are situated at the apex of alluvial fans and in the path of potential debris flows. In this study, we conducted simulations of debris flows to assess the risks in inhabited areas throughout the Philippines and validated the results in the field, focusing on the provinces of Pangasinan and Aurora as primary examples. Watersheds that drain in an alluvial fan using a 10-m resolution Synthetic Aperture Radar (SAR)-derived Digital Elevation Model (DEM) was first delineated, and then a 1 in 100-year rain return rainfall scenario for the watershed was used to simulate debris flows using FLO-2D, a flood-routing software. The resulting simulations were used to generate debris flow hazard maps which are consistent with danger zones in alluvial fans delineated previously from satellite imagery and available DEMs. The

  12. An Experimental Study of Flow Separation over 2D Transverse Grooves

    NASA Astrophysics Data System (ADS)

    Jones, Emily; Lang, Amy; Afroz, Farhana; Wheelus, Jennifer; Smith, Drew

    2011-11-01

    A shark's scales help to reduce drag over its body by controlling boundary layer separation over its skin. It is theorized that the scales bristle when encountering a reversing flow, thereby trapping vortices between the scales, creating a partial slip condition over the surface and inducing turbulence augmentation in the boundary layer. In an attempt to replicate and study these effects, a spinning cylinder was used in a water tunnel to induce separation over a flat plate with 2 millimeter square 2D transverse grooves. The results were compared to separation occurring over a flat plate without grooves using DPIV. The angular speed of the cylinder was varied. The observed delays in separation, changes in separation bubble shedding frequency and other effects upon the boundary layer are discussed.

  13. Micro PIV measurements of turbulent flow over 2D structured roughness

    NASA Astrophysics Data System (ADS)

    Hartenberger, Joel; Perlin, Marc

    2015-11-01

    We investigate the turbulent boundary layer over surfaces with 2D spanwise square and triangular protrusions having nominal heights of 100 - 300 microns for Reynolds numbers ranging from Reτ ~ 1500 through Reτ ~ 4500 using a high speed, high magnification imaging system. Micro PIV analysis gives finely resolved velocity fields of the flow (on the order of 10 microns between vectors) enabling a detailed look at the inner region as well as the flow in the immediate vicinity of the roughness elements. Additionally, planar PIV with lower resolution is performed to capture the remainder of the boundary layer to the freestream flow. Varying the streamwise distance between individual roughness elements from one to ten times the nominal heights allows investigation of k-type and d-type roughness in both the transitionally rough and fully rough regimes. Preliminary results show a shift in the mean velocity profile similar to the results of previous studies. Turbulent statistics will be presented also. The authors would like to acknowledge the support of NAVSEA which funded this project through the Naval Engineering Education Center (NEEC).

  14. A friction to flow constitutive law and its application to a 2-D modeling of earthquakes

    NASA Astrophysics Data System (ADS)

    Shimamoto, Toshihiko; Noda, Hiroyuki

    2014-11-01

    Establishment of a constitutive law from friction to high-temperature plastic flow has long been a challenging task for solving problems such as modeling earthquakes and plate interactions. Here we propose an empirical constitutive law that describes this transitional behavior using only friction and flow parameters, with good agreements with experimental data on halite shear zones. The law predicts steady state and transient behaviors, including the dependence of the shear resistance of fault on slip rate, effective normal stress, and temperature. It also predicts a change in velocity weakening to velocity strengthening with increasing temperature, similar to the changes recognized for quartz and granite gouge under hydrothermal conditions. A slight deviation from the steady state friction law due to the involvement of plastic deformation can cause a large change in the velocity dependence. We solved seismic cycles of a fault across the lithosphere with the law using a 2-D spectral boundary integral equation method, revealing dynamic rupture extending into the aseismic zone and rich evolution of interseismic creep including slow slip prior to earthquakes. Seismic slip followed by creep is consistent with natural pseudotachylytes overprinted with mylonitic deformation. Overall fault behaviors during earthquake cycles are insensitive to transient flow parameters. The friction-to-flow law merges "Christmas tree" strength profiles of the lithosphere and rate dependency fault models used for earthquake modeling on a unified basis. Strength profiles were drawn assuming a strain rate for the flow regime, but we emphasize that stress distribution evolves reflecting the fault behavior. A fault zone model was updated based on the earthquake modeling.

  15. Altered NK Cell Development and Enhanced NK Cell-Mediated Resistance to MCMV in NKG2D-Deficient Mice

    PubMed Central

    Zafirova, Biljana; Mandarić, Sanja; Antulov, Ronald; Krmpotić, Astrid; Jonsson, Helena; Yokoyama, Wayne M.; Jonjić, Stipan; Polić, Bojan

    2009-01-01

    Summary NKG2D is a potent activating receptor on NK cells which acts as a molecular sensor for stressed cells expressing NKG2D ligands such as infected or tumor transformed cells. Although NKG2D is expressed on NK cell precursors, its role in NK cell development is still not known. We have generated NKG2D-deficient mice by targeting the Klrk1 locus. Here we provide evidence for an important regulatory role of NKG2D in the development of NK cells. The absence of NKG2D causes faster division of NK cells, perturbation in size of some NK cell subpopulations and their augmented sensitivity to apoptosis. As expected, NKG2D−/− NK cells are less responsive to tumor targets expressing NKG2D ligands. NKG2D−/− mice, however, show an enhanced NK cell-mediated resistance to MCMV infection as a consequence of NK cell dysregulation. Altogether, these findings provide evidence for yet unknown regulatory function of NKG2D in NK cell physiology. PMID:19631564

  16. Heat Flow Partitioning Between Continents and Oceans - from 2D to 3D

    NASA Astrophysics Data System (ADS)

    Moresi, L. N.; Cooper, C. M.; Lenardic, A.

    2010-12-01

    Scalings derived from thermal network theory explain how the presence of continents can influence the Earth’s overall heat loss. Intuitively, it may seem that increasing the proportion of a planet’s surface area covered by continents would decrease the efficiency of heat transfer given that continents do not participate in convective overturn. However, this ignores the potential feedback between the insulating effect of continents and the temperature-dependent viscosity of the mantle (Lenardic et al, 2005, Cooper et al, 2007). When this feedback is considered, a clear regime exists in which the partial stagnation and insulation of the surface by buoyant continental crust can lead to an increase in heat flow compared to the uninsulated case. The numerical results used to verify the scalings have mostly been conducted in two dimensions in order to cover a very wide range of Rayleigh number, fraction of continental coverage, and continental thickness. However as more recent results show that the configuration of the crust also plays a role in determining the heat flow partitioning and global heat flow (See Lenardic et al, “Continents, Super-Continents, Mantle Thermal Mixing, and Mantle Thermal Isolation” in this session), we have begun to repeat this exhaustive and exhausting 2D study in 3D. Cooper, C.M., A. Lenardic, and L.-N. Moresi "Effects of continental insulation and the partioning of heat producing elements on the Earth's heat loss." Geophys. Res. Lett., 33 ,10.1029, 2006. Lenardic, A., L.-N. Moresi, A.M. Jellinek, and M. Manga "Continental insulation, mantle cooling, and the surface area of oceans and continents." Earth Planet. Sci. Lett., 234 ,317-333, 2005.

  17. 4-D flow magnetic resonance imaging: blood flow quantification compared to 2-D phase-contrast magnetic resonance imaging and Doppler echocardiography

    PubMed Central

    Gabbour, Maya; Schnell, Susanne; Jarvis, Kelly; Robinson, Joshua D.; Markl, Michael

    2015-01-01

    Background Doppler echocardiography (echo) is the reference standard for blood flow velocity analysis, and two-dimensional (2-D) phase-contrast magnetic resonance imaging (MRI) is considered the reference standard for quantitative blood flow assessment. However, both clinical standard-of-care techniques are limited by 2-D acquisitions and single-direction velocity encoding and may make them inadequate to assess the complex three-dimensional hemodynamics seen in congenital heart disease. Four-dimensional flow MRI (4-D flow) enables qualitative and quantitative analysis of complex blood flow in the heart and great arteries. Objectives The objectives of this study are to compare 4-D flow with 2-D phase-contrast MRI for quantification of aortic and pulmonary flow and to evaluate the advantage of 4-D flow-based volumetric flow analysis compared to 2-D phase-contrast MRI and echo for peak velocity assessment in children and young adults. Materials and methods Two-dimensional phase-contrast MRI of the aortic root, main pulmonary artery (MPA), and right and left pulmonary arteries (RPA, LPA) and 4-D flow with volumetric coverage of the aorta and pulmonary arteries were performed in 50 patients (mean age: 13.1±6.4 years). Four-dimensional flow analyses included calculation of net flow and regurgitant fraction with 4-D flow analysis planes similarly positioned to 2-D planes. In addition, 4-D flow volumetric assessment of aortic root/ascending aorta and MPA peak velocities was performed and compared to 2-D phase-contrast MRI and echo. Results Excellent correlation and agreement were found between 2-D phase-contrast MRI and 4-D flow for net flow (r=0.97, P<0.001) and excellent correlation with good agreement was found for regurgitant fraction (r= 0.88, P<0.001) in all vessels. Two-dimensional phase-contrast MRI significantly underestimated aortic (P= 0.032) and MPA (P<0.001) peak velocities compared to echo, while volumetric 4-D flow analysis resulted in higher (aortic: P=0

  18. Full 2D observation of water surface elevation from SWOT under different flow conditions

    NASA Astrophysics Data System (ADS)

    Domeneghetti, Alessio; Schumann, Guy; Rui, Wei; Durand, Michael; Pavelsky, Tamlin

    2016-04-01

    The upcoming Surface Water and Ocean Topography (SWOT) satellite mission is a joint project of NASA, Centre National d'Etudes Spatiales (CNES, France), the Canadian Space Agency, and the Space Agency of the UK that will provide a first global, high-resolution observation of ocean and terrestrial water surface heights. Characterized by an observation swath of 120 km and an orbit repeat interval of about 21 days, SWOT will provide unprecedented bi-dimensional observations of rivers wider than 50-100 m. Despite many research activities that have investigated potential uses of remotely sensed data from SWOT, potentials and limitations of the spatial observations provided by the satellite mission for flood modeling still remain poorly understood and investigated. In this study we present a first analysis of the spatial observation of water surface elevation that is expected from SWOT for a 140 km reach of the middle-lower portion of the Po River, in Northern Italy. The river stretch is characterized by a main channel varying from 200-500 m in width and a floodplain that can be as wide as 5 km and that is delimited by a system of major embankments. The reconstruction of the hydraulic behavior of the Po River is performed by means of a quasi-2d model built with detailed topographic and bathymetric information (LiDAR, 2 m resolution), while the simulation of the spatial observation sensed by SWOT is performed with a SWOT simulator that mimics the satellite sensor characteristics. Referring to water surface elevations associated with different flow conditions (maximum, minimum and average flow reproduced by means of the quasi-2d numerical model) this work provides a first characterization of the spatial observations provided by SWOT and highlights the strengths and limitations of the expected products. By referring to a real river reach the analysis provides a credible example of the type of spatial observations that will be available after launch of SWOT and offers a first

  19. Enhanced photon absorption in spiral nanostructured solar cells using layered 2D materials.

    PubMed

    Tahersima, Mohammad H; Sorger, Volker J

    2015-08-28

    Recent investigations of semiconducting two-dimensional (2D) transition metal dichalcogenides have provided evidence for strong light absorption relative to its thickness attributed to high density of states. Stacking a combination of metallic, insulating, and semiconducting 2D materials enables functional devices with atomic thicknesses. While photovoltaic cells based on 2D materials have been demonstrated, the reported absorption is still just a few percent of the incident light due to their sub-wavelength thickness leading to low cell efficiencies. Here we show that taking advantage of the mechanical flexibility of 2D materials by rolling a molybdenum disulfide (MoS(2))/graphene (Gr)/hexagonal boron nitride stack to a spiral solar cell allows for optical absorption up to 90%. The optical absorption of a 1 μm long hetero-material spiral cell consisting of the aforementioned hetero stack is about 50% stronger compared to a planar MoS(2) cell of the same thickness; although the volumetric absorbing material ratio is only 6%. A core-shell structure exhibits enhanced absorption and pronounced absorption peaks with respect to a spiral structure without metallic contacts. We anticipate these results to provide guidance for photonic structures that take advantage of the unique properties of 2D materials in solar energy conversion applications.

  20. T cell triggering: insights from 2D kinetics analysis of molecular interactions

    NASA Astrophysics Data System (ADS)

    Zarnitsyna, Veronika; Zhu, Cheng

    2012-08-01

    Interaction of the T cell receptor (TCR) with pathogen-derived peptide presented by the major histocompatibility complex (pMHC) molecule is central to adaptive immunity as it initiates intracellular signaling to trigger T cell response to infection. Kinetic parameters of this interaction have been under intensive investigation for more than two decades using soluble pMHCs and/or TCRs with at least one of them in the solution (three-dimensional (3D) methods). Recently, several techniques have been developed to enable kinetic analysis on live T cells with pMHCs presented by surrogate antigen presenting cells (APCs) or supported planar lipid bilayers (two-dimensional (2D) methods). Comparison of 2D versus 3D parameters reveals drastic differences with broader ranges of 2D affinities and on-rates and orders of magnitude faster 2D off-rates for functionally distinct pMHCs. Here we review new 2D data and discuss how it may impact previously developed models of T cell discrimination between pMHCs of different potencies.

  1. L-plastin is involved in NKG2D recruitment into lipid rafts and NKG2D-mediated NK cell migration.

    PubMed

    Serrano-Pertierra, Esther; Cernuda-Morollón, Eva; Brdička, Tomáš; Hoøejši, Václav; López-Larrea, Carlos

    2014-09-01

    Membrane rafts are microdomains of the plasma membrane that have multiple biological functions. The involvement of these structures in the biology of T cells, namely in signal transduction by the TCR, has been widely studied. However, the role of membrane rafts in immunoreceptor signaling in NK cells is less well known. We studied the distribution of the activating NKG2D receptor in lipid rafts by isolating DRMs in a sucrose density gradient or by raft fractionation by β-OG-selective solubility in the NKL cell line. We found that the NKG2D-DAP10 complex and pVav are recruited into rafts upon receptor stimulation. Qualitative proteomic analysis of these fractions showed that the actin cytoskeleton is involved in this process. In particular, we found that the actin-bundling protein L-plastin plays an important role in the clustering of NKG2D into lipid rafts. Moreover, coengagement of the inhibitory receptor NKG2A partially disrupted NKG2D recruitment into rafts. Furthermore, we demonstrated that L-plastin participates in NKG2D-mediated inhibition of NK cell chemotaxis.

  2. Solution-Adaptive Program for Computing 2D/Axi Viscous Flow

    NASA Technical Reports Server (NTRS)

    Wood, William A.

    2003-01-01

    A computer program solves the Navier- Stokes equations governing the flow of a viscous, compressible fluid in an axisymmetric or two-dimensional (2D) setting. To obtain solutions more accurate than those generated by prior such programs that utilize regular and/or fixed computational meshes, this program utilizes unstructured (that is, irregular triangular) computational meshes that are automatically adapted to solutions. The adaptation can refine to regions of high change in gradient or can be driven by a novel residual minimization technique. Starting from an initial mesh and a corresponding data structure, the adaptation of the mesh is controlled by use of minimization functional. Other improvements over prior such programs include the following: (1) Boundary conditions are imposed weakly; that is, following initial specification of solution values at boundary nodes, these values are relaxed in time by means of the same formulations as those used for interior nodes. (2) Eigenvalues are limited in order to suppress expansion shocks. (3) An upwind fluctuation-splitting distribution scheme applied to inviscid flux requires fewer operations and produces less artificial dissipation than does a finite-volume scheme, leading to greater accuracy of solutions.

  3. Titan2D Based Pyroclastic Flows Hazard Maps for Santa Ana Volcano, El Salvador

    NASA Astrophysics Data System (ADS)

    Bajo, J. V.; Martinez-Hackert, B.; Escobar, C. D.; Gutierrez, R. E.

    2009-05-01

    Santa Ana Volcano is located in the Apaneca Volcanic Field located to the west of El Salvador, Central America. It is one the six active volcanoes monitor by the Servicios Nacionales de Estudios Territoriales (SNET) in El Salvador, out of twenty that are considered active in this small country by Smithsonian definition. The Santa Ana Volcano is surrounded by rural communities in its proximal areas and in its close distal areas by the second largest city of the country. On October 1st 2005, after a few months of increased fumarolic and seismic activity, it erupted generating a 10 km high steam and ash plume, reportedly seen by some aircraft and estimated using photography by SNET members. Ash was deposited to the west, north-west part of the country, following typical wind pattern for the region, as well as small pyroclastic flows and major lahars in its eastern part. Coffee plantations were lost, as was some crop of coffee in the following season. However, to the west the ash fertilized the land and resulted in an enhanced harvest of coffee beans. Only 2 people were killed from the Blast, thanks to the auto evacuation of proximal communities. Whilst the last eruption had a relatively low human life toll, a stronger eruption spells havoc almost certainly for the region. At this moment no exhaustive study and understanding exists of the pyroclastic flows generated by the Santa Ana Volcano nor a map for this particular hazard. This study proposes the use of Titan2D for those two purposes, using a DEM generated by the SNET using topographic maps as well as DEMs generated using Advanced Spaceborne Thermal Emission and Reflection Radiometer Images (ASTER).

  4. Increased serum NKG2D-ligands and downregulation of NKG2D in peripheral blood NK cells of patients with major burns.

    PubMed

    Haik, Josef; Nardini, Gil; Goldman, Noga; Galore-Haskel, Gilli; Harats, Moti; Zilinsky, Isaac; Weissman, Oren; Schachter, Jacob; Winkler, Eyal; Markel, Gal

    2016-01-19

    Immune suppression following major thermal injury directly impacts the recovery potential. Limited data from past reports indicate that natural killer cells might be suppressed due to a putative soluble factor that has remained elusive up to date. Here we comparatively study cohorts of patients with Major and Non-Major Burns as well as healthy donors. MICB and ULBP1 are stress ligands of NKG2D that can be induced by heat stress. Remarkably, serum concentration levels of MICB and ULBP1 are increased by 3-fold and 20-fold, respectively, already within 24h post major thermal injury, and are maintained high for 28 days. In contrast, milder thermal injuries do not similarly enhance the serum levels of MICB and ULBP1. This kinetics coincides with a significant downregulation of NKG2D expression among peripheral blood NK cells. Downregulation of NKG2D by high concentration of soluble MICB occurs in cancer patients and during normal pregnancy due to over production by cancer cells or extravillous trophoblasts, respectively, as an active immune-evasion mechanism. In burn patients this seems an incidental outcome of extensive thermal injury, leading to reduced NKG2D expression. Enhanced susceptibility of these patients to opportunistic viral infections, particularly herpes viruses, could be explained by the reduced NKG2D expression. Further studies are warranted for translation into innovative diagnostic or therapeutic technologies. PMID:26745675

  5. Activated and expanded natural killer cells target osteosarcoma tumor initiating cells in an NKG2D-NKG2DL dependent manner.

    PubMed

    Fernández, L; Valentín, J; Zalacain, M; Leung, W; Patiño-García, A; Pérez-Martínez, A

    2015-11-01

    Current therapies fail to cure most metastatic or recurrent bone cancer. We explored the efficacy and the pathways involved in natural killer (NK) cells' elimination of osteosarcoma (OS) cells, including tumor initiating cells (TICs), which are responsible for chemotherapy resistance, recurrence, and metastasis. The expression of ligands for NK cell receptors was studied in primary OS cell lines by flow cytometry. In vitro cytotoxicity of activated and expanded NK (NKAE) cells against OS was tested, and the pathways involved explored by using specific antibody blockade. NKAE cells' ability to target OS TICs was analyzed by flow cytometry and sphere formation assays. Spironolactone (SPIR) was tested for its ability to increase OS cells' susceptibility to NK cell lysis in vitro and in vivo. We found OS cells were susceptible to NKAE cells' lysis both in vivo and in vitro, and this cytolytic activity relied on interaction between NKG2D receptor and NKG2D ligands (NKG2DL). SPIR increased OS cells' susceptibility to lysis by NKAE cells, and could shrink the OS TICs. Our results show NKAE cells target OS cells including the TICs compartment, supporting the use of NK-cell based immunotherapies for OS.

  6. Approaches to Modeling Coupled Flow and Reaction in a 2-D Cementation Experiment

    SciTech Connect

    Steefel, Carl; Cochepin, B.; Trotignon, L.; Bildstein, O.; Steefel, C.; Lagneau, V.; van der Lee, J.

    2008-04-01

    Porosity evolution at reactive interfaces is a key process that governs the evolution and performances of many engineered systems that have important applications in earth and environmental sciences. This is the case, for example, at the interface between cement structures and clays in deep geological nuclear waste disposals. Although in a different transport regime, similar questions arise for permeable reactive barriers used for biogeochemical remediation in surface environments. The COMEDIE project aims at investigating the coupling between transport, hydrodynamics and chemistry when significant variations of porosity occur. The present work focuses on a numerical benchmark used as a design exercise for the future COMEDIE-2D experiment. The use of reactive transport simulation tools like Hytec and Crunch provides predictions of the physico-chemical evolutions that are expected during the future experiments in laboratory. Focus is given in this paper on the evolution during the simulated experiment of precipitate, permeability and porosity fields. A first case is considered in which the porosity is constant. Results obtained with Crunch and Hytec are in relatively good agreement. Differences are attributable to the models of reactive surface area taken into account for dissolution/precipitation processes. Crunch and Hytec simulations taking into account porosity variations are then presented and compared. Results given by the two codes are in qualitative agreement, with differences attributable in part to the models of reactive surface area for dissolution/precipitation processes. As a consequence, the localization of secondary precipitates predicted by Crunch leads to lower local porosities than for predictions obtained by Hytec and thus to a stronger coupling between flow and chemistry. This benchmark highlights the importance of the surface area model employed to describe systems in which strong porosity variations occur as a result of dissolution

  7. Endothelial cell activation and proliferation modulate NKG2D activity by regulating MICA expression and shedding.

    PubMed

    Chauveau, Annabelle; Tonnerre, Pierre; Pabois, Angélique; Gavlovsky, Pierre-Jean; Chatelais, Mathais; Coupel, Stéphanie; Charreau, Béatrice

    2014-01-01

    MICA are major histocompatibility complex class I-related molecules, expressed by endothelial cells (ECs), that may be targets for alloantibodies and NKG2D-expressing natural killer (NK) and T effector cells in organ allografts. This study shows that basal levels of MICA expressed on vascular ECs is sufficient to functionally modulate the expression and activity of the immunoreceptor NKG2D in allogeneic NK cells. We found that MICA expression is differentially regulated at the EC surface in response to cytokines. TNFα upregulates MICA while IFNγ significantly decreases MICA at the EC surface. Both cytokines induce the release of soluble MICA by ECs. Modulation of NKG2D correlates with the MICA level on the EC surface. Glycosylation and metalloproteinase activities account for major post-transcriptional mechanisms controlling MICA level and the function in ECs. Our results indicate that, in addition to the NFκB pathway, the mitogen-activated protein kinase pathways JNK, ERK1/2 and p38 are key signaling pathways in the control of MICA by the cytokines. Finally, we show that EC proliferation mediated by FGF-2 or wound healing increases the MICA level. Together, our data suggest that inflammation and proliferation regulate endothelial MICA expression and shedding, enabling ECs to modulate NKG2D activity on effector NK and T cells, and provide further evidence of a role for ECs in immunoregulation.

  8. SIMULATIONS OF 2D AND 3D THERMOCAPILLARY FLOWS BY A LEAST-SQUARES FINITE ELEMENT METHOD. (R825200)

    EPA Science Inventory

    Numerical results for time-dependent 2D and 3D thermocapillary flows are presented in this work. The numerical algorithm is based on the Crank-Nicolson scheme for time integration, Newton's method for linearization, and a least-squares finite element method, together with a matri...

  9. Defining an optimal surface chemistry for pluripotent stem cell culture in 2D and 3D

    NASA Astrophysics Data System (ADS)

    Zonca, Michael R., Jr.

    Surface chemistry is critical for growing pluripotent stem cells in an undifferentiated state. There is great potential to engineer the surface chemistry at the nanoscale level to regulate stem cell adhesion. However, the challenge is to identify the optimal surface chemistry of the substrata for ES cell attachment and maintenance. Using a high-throughput polymerization and screening platform, a chemically defined, synthetic polymer grafted coating that supports strong attachment and high expansion capacity of pluripotent stem cells has been discovered using mouse embryonic stem (ES) cells as a model system. This optimal substrate, N-[3-(Dimethylamino)propyl] methacrylamide (DMAPMA) that is grafted on 2D synthetic poly(ether sulfone) (PES) membrane, sustains the self-renewal of ES cells (up to 7 passages). DMAPMA supports cell attachment of ES cells through integrin beta1 in a RGD-independent manner and is similar to another recently reported polymer surface. Next, DMAPMA has been able to be transferred to 3D by grafting to synthetic, polymeric, PES fibrous matrices through both photo-induced and plasma-induced polymerization. These 3D modified fibers exhibited higher cell proliferation and greater expression of pluripotency markers of mouse ES cells than 2D PES membranes. Our results indicated that desirable surfaces in 2D can be scaled to 3D and that both surface chemistry and structural dimension strongly influence the growth and differentiation of pluripotent stem cells. Lastly, the feasibility of incorporating DMAPMA into a widely used natural polymer, alginate, has been tested. Novel adhesive alginate hydrogels have been successfully synthesized by either direct polymerization of DMAPMA and methacrylic acid blended with alginate, or photo-induced DMAPMA polymerization on alginate nanofibrous hydrogels. In particular, DMAPMA-coated alginate hydrogels support strong ES cell attachment, exhibiting a concentration dependency of DMAPMA. This research provides a

  10. The MICA-129 dimorphism affects NKG2D signaling and outcome of hematopoietic stem cell transplantation

    PubMed Central

    Isernhagen, Antje; Malzahn, Dörthe; Viktorova, Elena; Elsner, Leslie; Monecke, Sebastian; von Bonin, Frederike; Kilisch, Markus; Wermuth, Janne Marieke; Walther, Neele; Balavarca, Yesilda; Stahl-Hennig, Christiane; Engelke, Michael; Walter, Lutz; Bickeböller, Heike; Kube, Dieter; Wulf, Gerald; Dressel, Ralf

    2015-01-01

    The MHC class I chain-related molecule A (MICA) is a highly polymorphic ligand for the activating natural killer (NK)-cell receptor NKG2D. A single nucleotide polymorphism causes a valine to methionine exchange at position 129. Presence of a MICA-129Met allele in patients (n = 452) undergoing hematopoietic stem cell transplantation (HSCT) increased the chance of overall survival (hazard ratio [HR] = 0.77, P = 0.0445) and reduced the risk to die due to acute graft-versus-host disease (aGVHD) (odds ratio [OR] = 0.57, P = 0.0400) although homozygous carriers had an increased risk to experience this complication (OR = 1.92, P = 0.0371). Overall survival of MICA-129Val/Val genotype carriers was improved when treated with anti-thymocyte globulin (HR = 0.54, P = 0.0166). Functionally, the MICA-129Met isoform was characterized by stronger NKG2D signaling, triggering more NK-cell cytotoxicity and interferon-γ release, and faster co-stimulation of CD8+ T cells. The MICA-129Met variant also induced a faster and stronger down-regulation of NKG2D on NK and CD8+ T cells than the MICA-129Val isoform. The reduced cell surface expression of NKG2D in response to engagement by MICA-129Met variants appeared to reduce the severity of aGVHD. PMID:26483398

  11. The MICA-129 dimorphism affects NKG2D signaling and outcome of hematopoietic stem cell transplantation.

    PubMed

    Isernhagen, Antje; Malzahn, Dörthe; Viktorova, Elena; Elsner, Leslie; Monecke, Sebastian; von Bonin, Frederike; Kilisch, Markus; Wermuth, Janne Marieke; Walther, Neele; Balavarca, Yesilda; Stahl-Hennig, Christiane; Engelke, Michael; Walter, Lutz; Bickeböller, Heike; Kube, Dieter; Wulf, Gerald; Dressel, Ralf

    2015-10-19

    The MHC class I chain-related molecule A (MICA) is a highly polymorphic ligand for the activating natural killer (NK)-cell receptor NKG2D. A single nucleotide polymorphism causes a valine to methionine exchange at position 129. Presence of a MICA-129Met allele in patients (n = 452) undergoing hematopoietic stem cell transplantation (HSCT) increased the chance of overall survival (hazard ratio [HR] = 0.77, P = 0.0445) and reduced the risk to die due to acute graft-versus-host disease (aGVHD) (odds ratio [OR] = 0.57, P = 0.0400) although homozygous carriers had an increased risk to experience this complication (OR = 1.92, P = 0.0371). Overall survival of MICA-129Val/Val genotype carriers was improved when treated with anti-thymocyte globulin (HR = 0.54, P = 0.0166). Functionally, the MICA-129Met isoform was characterized by stronger NKG2D signaling, triggering more NK-cell cytotoxicity and interferon-γ release, and faster co-stimulation of CD8(+) T cells. The MICA-129Met variant also induced a faster and stronger down-regulation of NKG2D on NK and CD8(+) T cells than the MICA-129Val isoform. The reduced cell surface expression of NKG2D in response to engagement by MICA-129Met variants appeared to reduce the severity of aGVHD.

  12. Pattern formation in 2D flow of non-Newtonian fluids

    NASA Astrophysics Data System (ADS)

    Shelley, Michael; Ljubinko; Kondic; Palffy-Muhoray, Peter

    1997-03-01

    We explore the dynamics of the interface between a gas and a non-Newtonian fluid in a Hele-Shaw cell. If gas expands into fluid, the interface is unstable (Saffman-Taylor instability). This instability leads to viscous fingering for Newtonian fluids, but can produce dendritic morphology for non-Newtonian ones. Our analysis is based on the formulation of modified Darcy's law (Kondic, Palffy-Muhoray, and Shelley, Phys. Rev. E 54), 4536 R, 1996., where the problem reduces to nonlinear boundary value problem for pressure field in the fluid. We perform full numerical simulation of the time evolution of the interface. In the flow regime where elastic effects are negligible, it is found that shear-thinning character of the fluid considerably influences the morphology of the interface. We hope to understand experimentally observed dendritic structure, which also appears in many related physical problems, such as directional solidification.

  13. Towards 2D field-flow fractionation - Vector separation over slanted open cavities

    NASA Astrophysics Data System (ADS)

    Bernate, Jorge A.; Yang, Mengfei; Zhao, Hong; Risbud, Sumedh; Paul, Colin; Dallas, Matthew; Konstantopoulos, Konstantinos; Drazer, German; Shaqfeh, Eric S. G.

    2013-11-01

    Planar microfluidic platforms for vector chromatography, in which different species fan out in different directions and can be continuously sorted, are particularly promising for the high throughput separation of multicomponent mixtures. We carry out a computational study of the vector separation of dilute suspensions of rigid and flexible particles transported by a pressure-driven flow over an array of slanted open cavities. The numerical scheme is based on a Stokes flow boundary integral equation method. The simulations are performed in a periodic system without lateral confinement, relevant to microfluidic devices with negligible recirculation in the main channel. We study the deflection of rigid spherical particles, of flexible capsules as a model of white and red blood cells, and of rigid discoidal particles as a model of platelets. We characterize the deflection of different particles as a function of their size, shape, shear elasticity, their release position, and the geometric parameters of the channel. The simulations provide insight into the separation mechanism and allow the optimization of specific devices depending on the application. Good agreement with experiments is observed.

  14. T Cells Engineered With Chimeric Antigen Receptors Targeting NKG2D Ligands Display Lethal Toxicity in Mice.

    PubMed

    VanSeggelen, Heather; Hammill, Joanne A; Dvorkin-Gheva, Anna; Tantalo, Daniela G M; Kwiecien, Jacek M; Denisova, Galina F; Rabinovich, Brian; Wan, Yonghong; Bramson, Jonathan L

    2015-10-01

    Ligands for the NKG2D receptor are overexpressed on tumors, making them interesting immunotherapy targets. To assess the tumoricidal properties of T cells directed to attack NKG2D ligands, we engineered murine T cells with two distinct NKG2D-based chimeric antigen receptors (CARs): (i) a fusion between the NKG2D receptor and the CD3ζ chain and (ii) a conventional second-generation CAR, where the extracellular domain of NKG2D was fused to CD28 and CD3ζ. To enhance the CAR surface expression, we also engineered T cells to coexpress DAP10. In vitro functionality and surface expression levels of all three CARs was greater in BALB/c T cells than C57BL/6 T cells, indicating strain-specific differences. Upon adoptive transfer of NKG2D-CAR-T cells into syngeneic animals, we observed significant clinical toxicity resulting in morbidity and mortality. The severity of these toxicities varied between the CAR configurations and paralleled their in vitro NKG2D surface expression. BALB/c mice were more sensitive to these toxicities than C57BL/6 mice, consistent with the higher in vitro functionality of BALB/c T cells. Treatment with cyclophosphamide prior to adoptive transfer exacerbated the toxicity. We conclude that while NKG2D ligands may be useful targets for immunotherapy, the pursuit of NKG2D-based CAR-T cell therapies should be undertaken with caution. PMID:26122933

  15. VARSAT2D: Finite-element analysis of variably saturated two-dimensional flow. Information circular/1994

    SciTech Connect

    Nieber, J.L.; Friedel, M.J.; Munir, H.M.

    1994-01-01

    This information circular describes a computer program called VARSAT2D, a comprehensive unsaturated fluid flow simulator developed by the U.S. Bureau of Mines. VARSAT2D solves for either a vertical or horizontal, transient or steady-state solution in variably saturated, heterogeneous, anisotropic porous media using the Galerkin finite-element approach. Simplex triangular elements are used. Moisture retention characteristics are described by specifying either the Brooks and Corey, Brutsaert, or Van Genuchten power functions, with hysteresis described using the Maulem independent domain model. Boundary conditions may include any combination of time-varying seepage, pressure along solution domain boundaries and/or at internal node points, unit hydraulic gradient at the lower boundary, and a uniform source and/or sink. The program should be a welcome addition for mining and environmental hydrologists, researchers, and engineers interested in modeling unsaturated fluid flow.

  16. Renormalizability of the gradient flow in the 2D O(N) non-linear sigma model

    NASA Astrophysics Data System (ADS)

    Makino, Hiroki; Suzuki, Hiroshi

    2015-03-01

    It is known that the gauge field and its composite operators evolved by the Yang-Mills gradient flow are ultraviolet (UV) finite without any multiplicative wave function renormalization. In this paper, we prove that the gradient flow in the 2D O(N) non-linear sigma model possesses a similar property: The flowed N-vector field and its composite operators are UV finite without multiplicative wave function renormalization. Our proof in all orders of perturbation theory uses a (2+1)-dimensional field theoretical representation of the gradient flow, which possesses local gauge invariance without gauge field. As an application of the UV finiteness of the gradient flow, we construct the energy-momentum tensor in the lattice formulation of the O(N) non-linear sigma model that automatically restores the correct normalization and the conservation law in the continuum limit.

  17. Global 2D stability analysis of the cross lid-driven cavity flow with a streamfunction-vorticity approach

    NASA Astrophysics Data System (ADS)

    Gogoi, Bidyut B.

    2016-07-01

    We have recently analyzed the global two-dimensional (2D) stability of the staggered lid-driven cavity (LDC) flow with a higher order compact (HOC) approach. In the analysis, critical parameters are determined for both the parallel and anti-parallel motion of the lids and a detailed analysis has been carried out on either side of the critical values. In this article, we carry out an investigation of flow stabilities inside a two-sided cross lid-driven cavity with a pair of opposite lids moving in both parallel and anti-parallel directions. On discretization, the governing 2D Navier-Stokes (N-S) equations describing the steady flow and flow perturbations results in a generalized eigenvalue problem which is solved for determining the critical parameters on four different grids. Elaborate computation is performed for a wide range of Reynolds numbers (Re) on either side of the critical values in the range 200 ⩽ Re ⩽ 10000. For flows below the critical Reynolds number Rec, our numerical results are compared with established steady-state results and excellent agreement is obtained in all the cases. For Reynolds numbers above Rec, phase plane and spectral density analysis confirmed the existence of periodic, quasi-periodic, and stable flow patterns.

  18. A 2-D oscillating flow analysis in Stirling engine heat exchangers

    NASA Technical Reports Server (NTRS)

    Ahn, Kyung H.; Ibrahim, Mounir B.

    1991-01-01

    A two-dimensional oscillating flow analysis was conducted, simulating the gas flow inside Stirling heat exchangers. Both laminar and turbulent oscillating pipe flow were investigated numerically for Re(max) = 1920 (Va = 80), 10800 (Va = 272), 19300 (Va = 272), and 60800 (Va = 126). The results are compared with experimental results of previous investigators. Also, predictions of the flow regime on present oscillating flow conditions were checked by comparing velocity amplitudes and phase differences with those from laminar theory and quasi-steady profile. A high Reynolds number k-epsilon turbulence model was used for turbulent oscillating pipe flow. Finally, performance evaluation of the K-epsilon model was made to explore the applicability of quasi-steady turbulent models to unsteady oscillating flow analysis.

  19. A 2-D oscillating flow analysis in Stirling engine heat exchangers

    NASA Technical Reports Server (NTRS)

    Ahn, Kyung H.; Ibrahim, Mounir B.

    1991-01-01

    A two dimensional oscillating flow analysis was conducted, simulating the gas flow inside Stirling heat exchangers. Both laminar and turbulent oscillating pipe flow were investigated numerically for Re(max) = 1920 (Va = 80), 10800 (Va = 272), 19300 (Va = 272), and 60800 (Va = 126). The results are compared with experimental results of previous investigators. Also, predictions of the flow regime on present oscillating flow conditions were checked by comparing velocity amplitudes and phase differences with those from laminar theory and quasi-steady profile. A high Reynolds number k-epsilon turbulence model was used for turbulent oscillating pipe flow. Finally, performance evaluation of the K-epsilon model was made to explore the applicability of quasi-steady turbulent models to unsteady oscillating flow analysis.

  20. NKG2D-dependent cytotoxicity is controlled by ligand distribution in the target cell membrane

    PubMed Central

    Martinez, Emily; Brzostowski, Joseph A.; Long, Eric O.; Gross, Catharina C.

    2013-01-01

    While the importance of membrane microdomains in receptor-mediated activation of lymphocytes has been established, much less is known about the role of receptor ligand distribution on APC and target cells. Detergent-resistant membrane (DRM) domains, into which glycophosphatidylinositol (GPI)-linked proteins partition, are enriched in cholesterol and glycosphingolipids. ULBP1 is a GPI-linked ligand for natural cytotoxicity receptor NKG2D. To investigate how ULBP1 distribution on target cells affects NKG2D-dependent NK cell activation, we fused the extracellular domain of ULBP1 to the transmembrane domain of CD45. Introduction of this transmembrane domain eliminated the association of ULBP1 with the DRM fraction and caused a significant reduction of cytotoxicity and degranulation by NK cells. Clustering and lateral diffusion of ULBP1 was not affected by changes in the membrane anchor. These results show that the partitioning of receptor ligands in discrete membrane domains of target cells is an important determinant of NK cell activation. PMID:21464092

  1. 2D-CELL: image processing software for extraction and analysis of 2-dimensional cellular structures

    NASA Astrophysics Data System (ADS)

    Righetti, F.; Telley, H.; Leibling, Th. M.; Mocellin, A.

    1992-01-01

    2D-CELL is a software package for the processing and analyzing of photographic images of cellular structures in a largely interactive way. Starting from a binary digitized image, the programs extract the line network (skeleton) of the structure and determine the graph representation that best models it. Provision is made for manually correcting defects such as incorrect node positions or dangling bonds. Then a suitable algorithm retrieves polygonal contours which define individual cells — local boundary curvatures are neglected for simplicity. Using elementary analytical geometry relations, a range of metric and topological parameters describing the population are then computed, organized into statistical distributions and graphically displayed.

  2. Role of 2-D periodic symmetrical nanostructures in improving efficiency of thin film solar cells

    NASA Astrophysics Data System (ADS)

    Zhang, Wei; Jiang, Liyong; Li, Xiangyin

    2016-01-01

    We systematically investigated several different nanostructures in crystalline silicon (c-Si) thin film solar cells and then proposed a brand-new structure with two dimensional (2-D) periodic dielectric cylinders on the top and annular metal columns on bottom surface to enhance the optical harvesting. The periodic symmetrical nanostructures affect the solar cell efficiency due to the grating diffraction effect of dielectric columns and surface plasmon polaritons (SPPs) effect induced by metal nanostructures at the dielectric-metal interface. About 52.1% more optical absorption and 33.3% more power conversion efficiency are obtained, and the maximum short current reaches to 33.24 mA/cm2.

  3. Corticotropin-releasing factor induces immune escape of cervical cancer cells by downregulation of NKG2D.

    PubMed

    Song, Hyunkeun; Park, Hyunjin; Park, Gabin; Kim, Yeong Seok; Lee, Hyun-Kyung; Jin, Dong-Hoon; Kang, Hyung-Sik; Cho, Dae-Ho; Hur, Daeyoung

    2014-07-01

    Corticotropin-releasing factor (CRF), a coordinator of the body's responses to stress, is found in various cancer tissues and cell lines. However, the exact abilities of CRF to manipulate natural killer (NK) cells during immune response have not been studied. NKG2D is an activating receptor that is expressed on most NK and CD8+ T cells. MHC class I-related chain A (MICA) and UL16-binding protein (ULBP) 1, 2 and 3 are well-known ligands for NKG2D. In the present study, we reported our findings regarding the role of CRF in cervical cancer cell survival. Human cervical cancer cell line, HeLa cells, had significantly higher intracellular expression of UL16-binding protein 2 (ULBP2) following CRF treatment but had only slightly increased surface expression of ULBP2. Notably, MMPi (pan-metalloproteases inhibitor) blocked the release of ULBP2 molecules from the surface of HeLa cells. Furthermore, incubating NK cells with culture supernatants from CRF-treated HeLa cells, which contained soluble NKG2D ligand, reduced NK cell activity by decreasing surface expression of NKG2D. Collectively, downregulation of NKG2D by CRF-induced soluble NKG2D ligand provides a potential mechanism by which cervical cancer cells escape NKG2D-mediated attack under stress conditions.

  4. Levels of regulatory T cells CD69(+)NKG2D(+)IL-10(+) are increased in patients with autoimmune thyroid disorders.

    PubMed

    Rodríguez-Muñoz, Ana; Vitales-Noyola, Marlen; Ramos-Levi, Ana; Serrano-Somavilla, Ana; González-Amaro, Roberto; Marazuela, Mónica

    2016-03-01

    Regulatory T (Treg) cells play an important role in the pathogenesis of autoimmune thyroid disorders (AITD). New subsets of CD4(+)CD69(+) and CD4(+)NKG2D(+) T lymphocytes that behave as regulatory cells have been recently reported. The role of these immunoregulatory lymphocytes has not been previously explored in AITD. We analyzed by multi-parametric flow cytometry different Treg cell subsets in peripheral blood from 32 patients with AITD and 19 controls, and in thyroid tissue from seven patients. The suppressive activity was measured by an assay of inhibition of lymphocyte activation. We found a significant increased percentage of CD4(+)CD69(+)IL-10(+), CD4(+)CD69(+)NKG2D(+), and CD4(+)CD69(+)IL-10(+)NKG2D(+) cells, in peripheral blood from GD patients compared to controls. The increase in CD4(+)CD69(+)IL-10(+) and CD4(+)CD69(+)IL-10(+)NKG2D(+) T cells was especially remarkable in patients with active Graves' ophthalmopathy (GO), and a significant positive correlation between GO activity and CD4(+)CD69(+)IL-10(+) or CD4(+)CD69(+)IL-10(+)NKG2D(+) cells was also found. In addition, these cells were increased in patients with a more severe and/or prolonged disease. Thyroid from AITD patients showed an increased proportion of CD69(+) regulatory T cells subpopulations compared to autologous peripheral blood. The presence of CD69(+), NKG2D(+), and IL-10(+) cells was confirmed by immunofluorescence microscopy. In vitro functional assays showed that CD69(+) Treg cells exerted an important suppressive effect on the activation of T effector cells in controls, but not in AITD patients. Our findings suggest that the levels of CD69(+) regulatory lymphocytes are increased in AITD patients, but they are apparently unable to down-modulate the autoimmune response and tissue damage.

  5. 2-D Western blotting for evaluation of antibodies developed for detection of host cell protein.

    PubMed

    Berkelman, Tom; Harbers, Adriana; Bandhakavi, Sricharan

    2015-01-01

    Recombinant proteins generated for therapeutic use must be substantially free of residual host cell protein (HCP). The presence of host cell protein (HCP) is usually assayed by ELISA using a polyclonal antibody mixture raised against a population of proteins derived from the host cell background. This antibody should recognize as high a proportion as possible of the potential HCPs in a given sample. A recommended method for evaluating the assay involves two-dimensional electrophoretic separation followed by Western blotting.We present here a method using commercial anti-HCP antibody and samples derived from Chinese Hamster Ovary (CHO) cells. The 2-D electrophoresis procedure gives highly reproducible spot patterns and entire procedure can be completed in less than 2 days. Software analysis enables the straightforward generation of percent coverage values for the antibody when used to probe HCP-containing samples. PMID:25820736

  6. 2-D Western blotting for evaluation of antibodies developed for detection of host cell protein.

    PubMed

    Berkelman, Tom; Harbers, Adriana; Bandhakavi, Sricharan

    2015-01-01

    Recombinant proteins generated for therapeutic use must be substantially free of residual host cell protein (HCP). The presence of host cell protein (HCP) is usually assayed by ELISA using a polyclonal antibody mixture raised against a population of proteins derived from the host cell background. This antibody should recognize as high a proportion as possible of the potential HCPs in a given sample. A recommended method for evaluating the assay involves two-dimensional electrophoretic separation followed by Western blotting.We present here a method using commercial anti-HCP antibody and samples derived from Chinese Hamster Ovary (CHO) cells. The 2-D electrophoresis procedure gives highly reproducible spot patterns and entire procedure can be completed in less than 2 days. Software analysis enables the straightforward generation of percent coverage values for the antibody when used to probe HCP-containing samples.

  7. Selective targeting of IL-2 to NKG2D bearing cells for improved immunotherapy

    PubMed Central

    Ghasemi, Reza; Lazear, Eric; Wang, Xiaoli; Arefanian, Saeed; Zheleznyak, Alexander; Carreno, Beatriz M.; Higashikubo, Ryuji; Gelman, Andrew E.; Kreisel, Daniel; Fremont, Daved H.; Krupnick, Alexander Sasha

    2016-01-01

    Despite over 20 years of clinical use, IL-2 has not fulfilled expectations as a safe and effective form of tumour immunotherapy. Expression of the high affinity IL-2Rα chain on regulatory T cells mitigates the anti-tumour immune response and its expression on vascular endothelium is responsible for life threatening complications such as diffuse capillary leak and pulmonary oedema. Here we describe the development of a recombinant fusion protein comprised of a cowpox virus encoded NKG2D binding protein (OMCP) and a mutated form of IL-2 with poor affinity for IL-2Rα. This fusion protein (OMCP-mutIL-2) potently and selectively activates IL-2 signalling only on NKG2D-bearing cells, such as natural killer (NK) cells, without broadly activating IL-2Rα-bearing cells. OMCP-mutIL-2 provides superior tumour control in several mouse models of malignancy and is not limited by mouse strain-specific variability of NK function. In addition, OMCP-mutIL-2 lacks the toxicity and vascular complications associated with parental wild-type IL-2. PMID:27650575

  8. Selective targeting of IL-2 to NKG2D bearing cells for improved immunotherapy.

    PubMed

    Ghasemi, Reza; Lazear, Eric; Wang, Xiaoli; Arefanian, Saeed; Zheleznyak, Alexander; Carreno, Beatriz M; Higashikubo, Ryuji; Gelman, Andrew E; Kreisel, Daniel; Fremont, Daved H; Krupnick, Alexander Sasha

    2016-01-01

    Despite over 20 years of clinical use, IL-2 has not fulfilled expectations as a safe and effective form of tumour immunotherapy. Expression of the high affinity IL-2Rα chain on regulatory T cells mitigates the anti-tumour immune response and its expression on vascular endothelium is responsible for life threatening complications such as diffuse capillary leak and pulmonary oedema. Here we describe the development of a recombinant fusion protein comprised of a cowpox virus encoded NKG2D binding protein (OMCP) and a mutated form of IL-2 with poor affinity for IL-2Rα. This fusion protein (OMCP-mutIL-2) potently and selectively activates IL-2 signalling only on NKG2D-bearing cells, such as natural killer (NK) cells, without broadly activating IL-2Rα-bearing cells. OMCP-mutIL-2 provides superior tumour control in several mouse models of malignancy and is not limited by mouse strain-specific variability of NK function. In addition, OMCP-mutIL-2 lacks the toxicity and vascular complications associated with parental wild-type IL-2. PMID:27650575

  9. Comparison of 4D flow and 2D velocity-encoded phase contrast MRI sequences for the evaluation of aortic hemodynamics.

    PubMed

    Bollache, Emilie; van Ooij, Pim; Powell, Alex; Carr, James; Markl, Michael; Barker, Alex J

    2016-10-01

    The purpose of this study was to compare aortic flow and velocity quantification using 4D flow MRI and 2D CINE phase-contrast (PC)-MRI with either one-directional (2D-1dir) or three-directional (2D-3dir) velocity encoding. 15 healthy volunteers (51 ± 19 years) underwent MRI including (1) breath-holding 2D-1dir and (2) free breathing 2D-3dir PC-MRI in planes orthogonal to the ascending (AA) and descending (DA) aorta, as well as (3) free breathing 4D flow MRI with full thoracic aorta coverage. Flow quantification included the co-registration of the 2D PC acquisition planes with 4D flow MRI data, AA and DA segmentation, and calculation of AA and DA peak systolic velocity, peak flow and net flow volume for all sequences. Additionally, the 2D-3dir velocity taking into account the through-plane component only was used to obtain results analogous to a free breathing 2D-1dir acquisition. Good agreement was found between 4D flow and 2D-3dir peak velocity (differences = -3 to 6 %), peak flow (-7 %) and net volume (-14 to -9 %). In contrast, breath-holding 2D-1dir measurements exhibited indices significantly lower than free breathing 2D-3dir and 2D-1dir (differences = -35 to -7 %, p < 0.05). Finally, high correlations (r ≥ 0.97) were obtained for indices estimated with or without eddy current correction, with the lowest correlation observed for net volume. 4D flow and 2D-3dir aortic hemodynamic indices were in concordance. However, differences between respiration state and 2D-1dir and 2D-3dir measurements indicate that reference values should be established according to the PC-MRI sequence, especially for the widely used net flow (e.g. stroke volume in the AA). PMID:27435230

  10. Comparison of 4D flow and 2D velocity-encoded phase contrast MRI sequences for the evaluation of aortic hemodynamics.

    PubMed

    Bollache, Emilie; van Ooij, Pim; Powell, Alex; Carr, James; Markl, Michael; Barker, Alex J

    2016-10-01

    The purpose of this study was to compare aortic flow and velocity quantification using 4D flow MRI and 2D CINE phase-contrast (PC)-MRI with either one-directional (2D-1dir) or three-directional (2D-3dir) velocity encoding. 15 healthy volunteers (51 ± 19 years) underwent MRI including (1) breath-holding 2D-1dir and (2) free breathing 2D-3dir PC-MRI in planes orthogonal to the ascending (AA) and descending (DA) aorta, as well as (3) free breathing 4D flow MRI with full thoracic aorta coverage. Flow quantification included the co-registration of the 2D PC acquisition planes with 4D flow MRI data, AA and DA segmentation, and calculation of AA and DA peak systolic velocity, peak flow and net flow volume for all sequences. Additionally, the 2D-3dir velocity taking into account the through-plane component only was used to obtain results analogous to a free breathing 2D-1dir acquisition. Good agreement was found between 4D flow and 2D-3dir peak velocity (differences = -3 to 6 %), peak flow (-7 %) and net volume (-14 to -9 %). In contrast, breath-holding 2D-1dir measurements exhibited indices significantly lower than free breathing 2D-3dir and 2D-1dir (differences = -35 to -7 %, p < 0.05). Finally, high correlations (r ≥ 0.97) were obtained for indices estimated with or without eddy current correction, with the lowest correlation observed for net volume. 4D flow and 2D-3dir aortic hemodynamic indices were in concordance. However, differences between respiration state and 2D-1dir and 2D-3dir measurements indicate that reference values should be established according to the PC-MRI sequence, especially for the widely used net flow (e.g. stroke volume in the AA).

  11. Low NKp30, NKp46 and NKG2D expression and reduced cytotoxic activity on NK cells in cervical cancer and precursor lesions

    PubMed Central

    2009-01-01

    Background Persistent high risk HPV infection can lead to cervical cancer, the second most common malignant tumor in women worldwide. NK cells play a crucial role against tumors and virus-infected cells through a fine balance between activating and inhibitory receptors. Expression of triggering receptors NKp30, NKp44, NKp46 and NKG2D on NK cells correlates with cytolytic activity against tumor cells, but these receptors have not been studied in cervical cancer and precursor lesions. The aim of the present work was to study NKp30, NKp46, NKG2D, NKp80 and 2B4 expression in NK cells from patients with cervical cancer and precursor lesions, in the context of HPV infection. Methods NKp30, NKp46, NKG2D, NKp80 and 2B4 expression was analyzed by flow cytometry on NK cells from 59 patients with cervical cancer and squamous intraepithelial lesions. NK cell cytotoxicity was evaluated in a 4 hour CFSE/7-AAD flow cytometry assay. HPV types were identified by PCR assays. Results We report here for the first time that NK cell-activating receptors NKp30 and NKp46 are significantly down-regulated in cervical cancer and high grade squamous intraepithelial lesion (HGSIL) patients. NCRs down-regulation correlated with low cytolytic activity, HPV-16 infection and clinical stage. NKG2D was also down-regulated in cervical cancer patients. Conclusion Our results suggest that NKp30, NKp46 and NKG2D down-regulation represent an evasion mechanism associated to low NK cell activity, HPV-16 infection and cervical cancer progression. PMID:19531227

  12. An experimental study of flow separation over a flat plate with 2D transverse grooves

    NASA Astrophysics Data System (ADS)

    Jones, Emily Michelle

    Nature has long been an inspiration for research in engineering. In particular, the biological surfaces of aquatic swimmers have been studied for their potential as drag reducing surfaces. The hydrodynamic benefit of riblets, or grooves embedded parallel to the flow, which appear on many aquatic biological surfaces, have been well documented and implemented in practical engineering applications. However the skin of dolphins is embedded with grooves that run perpendicular to the flow of water over their bodies. It is theorized that the transverse grooves present on dolphin skin trap vortices between them, creating a partial slip condition over the surface and inducing turbulence augmentation in the boundary layer, thus controlling boundary layer separation over the dolphin's skin. Similarly, sharks are covered with scales that are flexible at the base and capable of bristling, forming grooves running transverse to the flow. It is theorized that the scales bristle when encountering a reversing flow, thereby trapping vortices between the scales and, similarly, delaying boundary layer separation. In an attempt to test this hypothesis and study these affects, a spinning cylinder was used in a water tunnel to induce separation over a flat plate with 2 mm, rectangular transverse grooves and sinusoidal grooves of similar scaling. The results were compared to tripped, turbulent boundary layer separation occurring over a flat plate without grooves using time-resolved particle image velocimetry. The strength of the adverse pressure gradient was varied, and the observed delay in flow separation and other affects upon the boundary layer are discussed.

  13. Wind-tunnel experiments of thermally-stratified turbulent boundary layer flow over a wall-mounted 2-D block

    NASA Astrophysics Data System (ADS)

    Zhang, Wei; Markfort, Corey; Porté-Agel, Fernando

    2014-05-01

    Turbulent boundary-layer flows over complex topography have been extensively studied in the atmospheric sciences and wind engineering communities. The upwind turbulence level, the atmospheric thermal stability and the shape of the topography as well as surface characteristics play important roles in turbulent transport of momentum and scalar fluxes. However, to the best of our knowledge, atmospheric thermal stability has rarely been taken into account in laboratory simulations, particularly in wind-tunnel experiments. Extension of such studies in thermally-stratified wind tunnels will substantially advance our understanding of thermal stability effects on the physics of flow over complex topography. Additionally, high-resolution experimental data can be used for development of new parameterization of surface fluxes and validation of numerical models such as Large-Eddy Simulation (LES). A series of experiments of neutral and thermally-stratified boundary-layer flows over a wall-mounted 2-D block were conducted at the Saint Anthony Falls Laboratory boundary-layer wind tunnel. The 2-D block, with a width to height ratio of 2:1, occupied the lowest 25% of the turbulent boundary layer. Stable and convective boundary layers were simulated by independently controlling the temperature of air flow, the test section floor, and the wall-mounted block surfaces. Measurements using high-resolution Particle Image Velocimetry (PIV), x-wire/cold-wire anemometry, thermal-couples and surface heat flux sensors were made to quantify the turbulent properties and surface fluxes in distinct macroscopic flow regions, including the separation/recirculation zones, evolving shear layer and the asymptotic far wake. Emphasis will be put on addressing thermal stability effects on the spatial distribution of turbulent kinetic energy (TKE) and turbulent fluxes of momentum and scalar from the near to far wake region. Terms of the TKE budget equation are also inferred from measurements and

  14. Numerical solution of 2D and 3D turbulent internal flow problems

    NASA Astrophysics Data System (ADS)

    Chen, Naixing; Xu, Yanji

    1991-08-01

    The paper describes a method for solving numerically two-dimensional or axisymmetric, and three-dimensional turbulent internal flow problems. The method is based on an implicit upwinding relaxation scheme with an arbitrarily shaped conservative control volume. The compressible Reynolds-averaged Navier-Stokes equations are solved with a two-equation turbulence model. All these equations are expressed by using a nonorthogonal curvilinear coordinate system. The method is applied to study the compressible internal flow in modern power installations. It has been observed that predictions for two-dimensional and three-dimensional channels show very good agreement with experimental results.

  15. Untangling tracer trajectories and clarifying coherence in 2D flows using braid theory

    NASA Astrophysics Data System (ADS)

    Filippi, Margaux; Atis, Séverine; Thiffeault, Jean-Luc; Budišić, Marko; Allshouse, Michael; Peacock, Thomas

    2014-11-01

    Interpreting ocean surface transport is crucial to many areas of oceanography, ranging from marine ecology to pollution management. To better understand surface mixing, we investigate a braid theory method to detect transport barriers bounding coherent structures in two-dimensional fluid flows. Whereas most existing techniques rely on an extensive spatiotemporal knowledge of the flow field, we seek to identify these structures from sparse data sets involving trajectories of a few tracer particles or floats. We present the results of model and laboratory experimental studies to test the robustness and applicability of the braid theory method, and discuss the potential applicability to oceanic data sets.

  16. Turbulence Measurements on a 2D NACA 0036 with Synthetic Jet Flow Control

    NASA Technical Reports Server (NTRS)

    Wilson, J. S.

    2006-01-01

    An active flow control experiment was conducted on a 2-ft chord NACA 0036 airfoil in a 3-ft by 4-ft Wind Tunnel at Re = 1 x 10(exp 6). The model was equipped with synthetic jet actuators at x/c = 0.30 and 0.65 that provided 120 Hz periodic excitation at a C(sub mu) 0.86% through 0.06-in wide slots. Three different slot con gurations were tested, including a baseline with no slots. Surface pressure data was collected to compare to previous tests and to combine with turbulence data to aid future CFD modeling efforts. Turbulence data, measured by hot-wire, was compared with and without flow control. Pressure data corroborates previous test data and provides more points for CFD validation. Hot-wire results showed ow control reduced the separated wake size and brought the high Reynolds stress shear layer closer to the airfoil surface. The position of this layer to the surface was altered more significantly than the magnitude of the peak stresses. Flow control was shown to increase turbulent energy in the attached boundary layer downstream of the slot but to have little effect upstream. These results provide further justification to continue assessing the potential of active flow control to reduce drag of helicopter airframe components.

  17. An investigation of design optimization using a 2-D viscous flow code with multigrid

    NASA Technical Reports Server (NTRS)

    Doria, Michael L.

    1990-01-01

    Computational fluid dynamics (CFD) codes have advanced to the point where they are effective analytical tools for solving flow fields around complex geometries. There is also a need for their use as a design tool to find optimum aerodynamic shapes. In the area of design, however, a difficulty arises due to the large amount of computer resources required by these codes. It is desired to streamline the design process so that a large number of design options and constraints can be investigated without overloading the system. There are several techniques which have been proposed to help streamline the design process. The feasibility of one of these techniques is investigated. The technique under consideration is the interaction of the geometry change with the flow calculation. The problem of finding the value of camber which maximizes the ratio of lift over drag for a particular airfoil is considered. In order to test out this technique, a particular optimization problem was tried. A NACA 0012 airfoil was considered at free stream Mach number of 0.5 with a zero angle of attack. Camber was added to the mean line of the airfoil. The goal was to find the value of camber for which the ratio of lift over drag is a maximum. The flow code used was FLOMGE which is a two dimensional viscous flow solver which uses multigrid to speed up convergence. A hyperbolic grid generation program was used to construct the grid for each value of camber.

  18. MEAN FLOW AND TURBULENCE MEASUREMENTS AROUND A 2-D ARRAY OF BUILDINGS IN A WIND TUNNEL

    EPA Science Inventory

    In order to predict the dispersion of harmful materials released in or near an urban environment, it is important to first understand the complex flow patterns which result from the interaction of the wind with buildings and, more commonly, clusters of buildings. Recent advanc...

  19. Lift force time delays on 2D and 3D wings in unsteady flows

    NASA Astrophysics Data System (ADS)

    Williams, David; Colling, Jesse; Quach, Vien; Colonius, Tim; Tadmor, Gilead

    2008-11-01

    Active flow control (AFC) used for enhancing the maneuverability of wings is usually applied during conditions of steady external flow. However, when the external flow is unsteady or the wing is maneuvering, then at least two time delays become important; namely, the time delay of the lift to changes in external flow, τf, and the time delay to changes in AFC actuation, τa. These time delays were measured in wind tunnel experiments using two- and three-dimensional wings in an oscillating freestream and with variable duty cycle actuation. Dimensionless freestream oscillation frequencies from k = 0.01 to k = 0.2 with amplitudes of 5 percent of the mean speed were used to characterize the system. As a demonstration of the important role of the two time constants, AFC is used to damp lift force oscillations occurring in an unsteady freestream using a feed forward control system. The instantaneous velocity provides input to a control algorithm which adjusts the duty cycle of the AFC actuator to suppress lift fluctuations.

  20. The NKG2D-IL-15 signaling pathway contributes to T-cell mediated pathology in inflammatory myopathies.

    PubMed

    Ruck, Tobias; Bittner, Stefan; Afzali, Ali Maisam; Göbel, Kerstin; Glumm, Sarah; Kraft, Peter; Sommer, Claudia; Kleinschnitz, Christoph; Preuße, Corinna; Stenzel, Werner; Wiendl, Heinz; Meuth, Sven G

    2015-12-22

    NKG2D is an activating receptor on T cells, which has been implicated in the pathogenesis of autoimmune diseases. T cells are critically involved in idiopathic inflammatory myopathies (IIM) and have been proposed as specific therapeutic targets. However, the mechanisms underlying T cell-mediated progressive muscle destruction in IIM remain to be elucidated. We here determined the involvement of the NKG2D - IL-15 signaling pathway. Primary human myoblasts expressed NKG2D ligands, which were further upregulated upon inflammatory stimuli. In parallel, shedding of the soluble NKG2D ligand MICA (sMICA) decreased upon inflammation potentially diminishing inhibition of NKG2D signaling. Membrane-related expression of IL-15 by myoblasts induced differentiation of naïve CD8+ T cells into highly activated, cytotoxic CD8+NKG2Dhigh T cells demonstrating NKG2D-dependent lysis of myoblasts in vitro. CD8+NKG2Dhigh T cell frequencies were increased in the peripheral blood of polymyositis (PM) patients and correlated with serum creatinine kinase concentrations, while serum sMICA levels were not significantly changed. In muscle biopsy specimens from PM patients expression of the NKG2D ligand MICA/B was upregulated, IL-15 was expressed by muscle cells, CD68+ macrophages as well as CD4+ T cells, and CD8+NKG2D+ cells were frequently detected within inflammatory infiltrates arguing for a local signaling circuit in the inflammatory muscle milieu. In conclusion, the NKG2D - IL-15 signaling pathway contributes to progressive muscle destruction in IIM potentially opening new therapeutic avenues. PMID:26646698

  1. The NKG2D – IL-15 signaling pathway contributes to T-cell mediated pathology in inflammatory myopathies

    PubMed Central

    Ruck, Tobias; Bittner, Stefan; Afzali, Ali Maisam; Göbel, Kerstin; Glumm, Sarah; Kraft, Peter; Sommer, Claudia; Kleinschnitz, Christoph; Preuβe, Corinna; Stenzel, Werner

    2015-01-01

    NKG2D is an activating receptor on T cells, which has been implicated in the pathogenesis of autoimmune diseases. T cells are critically involved in idiopathic inflammatory myopathies (IIM) and have been proposed as specific therapeutic targets. However, the mechanisms underlying T cell-mediated progressive muscle destruction in IIM remain to be elucidated. We here determined the involvement of the NKG2D – IL-15 signaling pathway. Primary human myoblasts expressed NKG2D ligands, which were further upregulated upon inflammatory stimuli. In parallel, shedding of the soluble NKG2D ligand MICA (sMICA) decreased upon inflammation potentially diminishing inhibition of NKG2D signaling. Membrane-related expression of IL-15 by myoblasts induced differentiation of naïve CD8+ T cells into highly activated, cytotoxic CD8+NKG2Dhigh T cells demonstrating NKG2D-dependent lysis of myoblasts in vitro. CD8+NKG2Dhigh T cell frequencies were increased in the peripheral blood of polymyositis (PM) patients and correlated with serum creatinine kinase concentrations, while serum sMICA levels were not significantly changed. In muscle biopsy specimens from PM patients expression of the NKG2D ligand MICA/B was upregulated, IL-15 was expressed by muscle cells, CD68+ macrophages as well as CD4+ T cells, and CD8+NKG2D+ cells were frequently detected within inflammatory infiltrates arguing for a local signaling circuit in the inflammatory muscle milieu. In conclusion, the NKG2D – IL-15 signaling pathway contributes to progressive muscle destruction in IIM potentially opening new therapeutic avenues. PMID:26646698

  2. Dataset of differentially expressed genes from SOX9 over-expressing NT2/D1 cells.

    PubMed

    Ludbrook, Louisa; Alankarage, Dimuthu; Bagheri-Fam, Stefan; Harley, Vincent

    2016-12-01

    The data presents the genes that are differentially up-regulated or down-regulated in response to SOX9 in a human Sertoli-like cell line, NT2/D1. The dataset includes genes that may be implicated in gonad development and are further explored in our associated article, "SOX9 Regulates Expression of the Male Fertility Gene Ets Variant Factor 5 (ETV5) during Mammalian Sex Development" (D. lankarage, R. Lavery, T. Svingen, S. Kelly, L.M. Ludbrook, S. Bagheri-Fam, et al., 2016) [1]. The necessity of SOX9 for male sex development is evident in instances where SOX9 is lost, as in 46, XY DSD where patients are sex reversed or in mouse knock-out models, where mice lacking Sox9 are sex reversed. Despite the crucial nature of this transcriptional activator, downstream target genes of SOX9 remain largely undiscovered. Here, we have utilized NT2/D1 cells to transiently over-express SOX9 and performed microarray analysis of the RNA. Microarray data are available in the ArrayExpress database (www.ebi.ac.uk/arrayexpress) under accession number E-MTAB-3378. PMID:27656672

  3. Critical role of the NKG2D receptor for NK cell-mediated control and immune escape of B-cell lymphoma.

    PubMed

    Belting, Lena; Hömberg, Nadine; Przewoznik, Margarethe; Brenner, Christoph; Riedel, Tanja; Flatley, Andrew; Polić, Bojan; Busch, Dirk H; Röcken, Martin; Mocikat, Ralph

    2015-09-01

    Little is known on the control of lymphomas by NK cells. Here, we study the role of the NK group 2D (NKG2D) receptor for the immunosurveillance of lymphoma. By using transplantable tumors as well as a λ-myc-transgenic model of endogenously arising lymphoma and NKG2D-deficient mice, we show that NK cells eliminate tumor cells in vivo after receiving two signals. One step involved the activation of NK cells giving rise to IFN-γ expression, which was effected by MHCI(low) tumor cells or DCs. However, this was necessary but not sufficient to mediate cytotoxicity. Triggering cytotoxicity additionally required a second step, which could be mediated by engagement of the NKG2D receptor. Thus, NKG2D-deficient NK cells could become activated in vivo, but they were not able to reject transplanted lymphomas or to degranulate in animals bearing autochthonous lymphomas. Tumor growth in NKG2D-deficient λ-myc-transgenic mice was significantly accelerated compared to NKG2D-competent animals. Whereas the latter developed tumors that lost expression of NKG2D ligands (NKG2D-L) in late disease stages, this did not occur in NKG2D-deficient mice. This indicates that NK cells and the NKG2D receptor play a role for control of lymphomas and that selection for NKG2D-L loss mutants provides a mechanism of tumor escape. PMID:26151313

  4. Critical role of the NKG2D receptor for NK cell-mediated control and immune escape of B-cell lymphoma.

    PubMed

    Belting, Lena; Hömberg, Nadine; Przewoznik, Margarethe; Brenner, Christoph; Riedel, Tanja; Flatley, Andrew; Polić, Bojan; Busch, Dirk H; Röcken, Martin; Mocikat, Ralph

    2015-09-01

    Little is known on the control of lymphomas by NK cells. Here, we study the role of the NK group 2D (NKG2D) receptor for the immunosurveillance of lymphoma. By using transplantable tumors as well as a λ-myc-transgenic model of endogenously arising lymphoma and NKG2D-deficient mice, we show that NK cells eliminate tumor cells in vivo after receiving two signals. One step involved the activation of NK cells giving rise to IFN-γ expression, which was effected by MHCI(low) tumor cells or DCs. However, this was necessary but not sufficient to mediate cytotoxicity. Triggering cytotoxicity additionally required a second step, which could be mediated by engagement of the NKG2D receptor. Thus, NKG2D-deficient NK cells could become activated in vivo, but they were not able to reject transplanted lymphomas or to degranulate in animals bearing autochthonous lymphomas. Tumor growth in NKG2D-deficient λ-myc-transgenic mice was significantly accelerated compared to NKG2D-competent animals. Whereas the latter developed tumors that lost expression of NKG2D ligands (NKG2D-L) in late disease stages, this did not occur in NKG2D-deficient mice. This indicates that NK cells and the NKG2D receptor play a role for control of lymphomas and that selection for NKG2D-L loss mutants provides a mechanism of tumor escape.

  5. CACNA2D2 promotes tumorigenesis by stimulating cell proliferation and angiogenesis.

    PubMed

    Warnier, M; Roudbaraki, M; Derouiche, S; Delcourt, P; Bokhobza, A; Prevarskaya, N; Mariot, P

    2015-10-16

    In the present study, we have assessed whether a putative calcium channel α2δ2 auxiliary subunit (CACNA2D2 gene) could be involved in prostate cancer (PCA) progression. We therefore carried out experiments to determine whether this protein is expressed in PCA LNCaP cells and in PCA tissues, and whether its expression may be altered during cancer development. In addition, we evaluated the influence on cell proliferation of overexpressing or downregulating this subunit. In vitro experiments show that α2δ2 subunit overexpression is associated with increased cell proliferation, alterations of calcium homeostasis and the recruitment of a nuclear factor of activated T-cells pathway. Furthermore, we carried out in vivo experiments on immuno-deficient nude mice in order to evaluate the tumorigenic potency of the α2δ2 subunit. We show that α2δ2-overexpressing PCA LNCaP cells are more tumorigenic than control LNCaP cells when injected into nude mice. In addition, gabapentin, a ligand of α2δ2, reduces tumor development in LNCaP xenografts. Finally, we show that the action of α2δ2 on tumor development occurs not only through a stimulation of proliferation, but also through a stimulation of angiogenesis, via an increased secretion of vascular endothelial growth factor in cells overexpressing α2δ2. PMID:25619833

  6. 2D models of gas flow and ice grain acceleration in Enceladus' vents using DSMC methods

    NASA Astrophysics Data System (ADS)

    Tucker, Orenthal J.; Combi, Michael R.; Tenishev, Valeriy M.

    2015-09-01

    The gas distribution of the Enceladus water vapor plume and the terminal speeds of ejected ice grains are physically linked to its subsurface fissures and vents. It is estimated that the gas exits the fissures with speeds of ∼300-1000 m/s, while the micron-sized grains are ejected with speeds comparable to the escape speed (Schmidt, J. et al. [2008]. Nature 451, 685-688). We investigated the effects of isolated axisymmetric vent geometries on subsurface gas distributions, and in turn, the effects of gas drag on grain acceleration. Subsurface gas flows were modeled using a collision-limiter Direct Simulation Monte Carlo (DSMC) technique in order to consider a broad range of flow regimes (Bird, G. [1994]. Molecular Gas Dynamics and the Direct Simulation of Gas Flows. Oxford University Press, Oxford; Titov, E.V. et al. [2008]. J. Propul. Power 24(2), 311-321). The resulting DSMC gas distributions were used to determine the drag force for the integration of ice grain trajectories in a test particle model. Simulations were performed for diffuse flows in wide channels (Reynolds number ∼10-250) and dense flows in narrow tubular channels (Reynolds number ∼106). We compared gas properties like bulk speed and temperature, and the terminal grain speeds obtained at the vent exit with inferred values for the plume from Cassini data. In the simulations of wide fissures with dimensions similar to that of the Tiger Stripes the resulting subsurface gas densities of ∼1014-1020 m-3 were not sufficient to accelerate even micron-sized ice grains to the Enceladus escape speed. In the simulations of narrow tubular vents with radii of ∼10 m, the much denser flows with number densities of 1021-1023 m-3 accelerated micron-sized grains to bulk gas speed of ∼600 m/s. Further investigations are required to understand the complex relationship between the vent geometry, gas source rate and the sizes and speeds of ejected grains.

  7. Experimental validation of a 2D overland flow model using high resolution water depth and velocity data

    NASA Astrophysics Data System (ADS)

    Cea, L.; Legout, C.; Darboux, F.; Esteves, M.; Nord, G.

    2014-05-01

    This paper presents a validation of a two-dimensional overland flow model using empirical laboratory data. Unlike previous publications in which model performance is evaluated as the ability to predict an outlet hydrograph, we use high resolution 2D water depth and velocity data to analyze to what degree the model is able to reproduce the spatial distribution of these variables. Several overland flow conditions over two impervious surfaces of the order of one square meter with different micro and macro-roughness characteristics are studied. The first surface is a simplified representation of a sinusoidal terrain with three crests and furrows, while the second one is a mould of a real agricultural seedbed terrain. We analyze four different bed friction parameterizations and we show that the performance of formulations which consider the transition between laminar, smooth turbulent and rough turbulent flow do not improve the results obtained with Manning or Keulegan formulas for rough turbulent flow. The simulations performed show that using Keulegan formula with a physically-based definition of the bed roughness coefficient, a two-dimensional shallow water model is able to reproduce satisfactorily the flow hydrodynamics. It is shown that, even if the resolution of the topography data and numerical mesh are high enough to include all the small scale features of the bed surface, the roughness coefficient must account for the macro-roughness characteristics of the terrain in order to correctly reproduce the flow hydrodynamics.

  8. 1α,25(OH)2D3 differentially regulates miRNA expression in human bladder cancer cells

    PubMed Central

    Ma, Yingyu; Hu, Qiang; Luo, Wei; Pratt, Rachel N.; Glenn, Sean T.; Liu, Song; Trump, Donald L.; Johnson, Candace S.

    2014-01-01

    Bladder cancer is the fourth most commonly diagnosed cancer in men and eighth leading cause of cancer-related death in the US. Epidemiological and experimental studies strongly suggest a role for 1α,25(OH)2D3 in cancer prevention and treatment. The antitumor activities of 1α,25(OH)2D3 are mediated by the induction of cell cycle arrest, apoptosis, differentiation and the inhibition of angiogenesis and metastasis. MiRNAs play important regulatory roles in cancer development and progression. However, the role of 1α,25(OH)2D3 in the regulation of miRNA expression and the potential impact in bladder cancer has not been investigated. Therefore, we studied 1α,25(OH)2D3-regulated miRNA expression profiles in human bladder cancer cell line 253J and the highly tumorigenic and metastatic derivative line 253J-BV by miRNA qPCR panels. 253 J and 253J-BV cells express endogenous vitamin D receptor (VDR) which can be further induced by 1α,25(OH)2D3. VDR target gene 24-hydroxylase was induced by 1α,25(OH)2D3 in both cell lines, indicating functional 1α,25(OH)2D3 signaling. The miRNA qPCR panel assay results showed that 253J and 253J-BV cells have distinct miRNA expression profiles. Further, 1α,25(OH)2D3 differentially regulated miRNA expression profiles in 253J and 253 J-BV cells in a dynamic manner. Pathway analysis of the miRNA target genes revealed distinct patterns of contribution to the molecular functions and biological processes in the two cell lines. In conclusion, 1α,25(OH)2D3 differentially regulates the expression of miRNAs, which may contribute to distinct biological functions, in human bladder 253J and 253J-BV cells. PMID:25263658

  9. Improved treatment of asthenosphere flow and melting in 2D numerical solutions for continental rifting: embedded vs nested modeling approaches.

    NASA Astrophysics Data System (ADS)

    de Monserrat, Albert; Morgan, Jason P.; Taramón, Jorge M.; Hall, Robert

    2016-04-01

    This work focuses on improving current 2D numerical approaches to modeling the boundary conditions associated with computing accurate deformation and melting associated with continental rifting. Recent models primarily use far-field boundary conditions that have been used for decades with little assessment of their effects on asthenospheric flow beneath the rifting region. All are extremely oversimplified. All are likely to significantly shape the pattern of asthenospheric flow beneath the stretching lithosphere which is associated with pressure-release melting and rift volcanism. The choice of boundary conditions may lead to different predictions of asthenospheric flow and melting associated with lithospheric stretching and breakup. We also find that they may affect the mode of crustal stretching. Here we discuss a suite of numerical experiments using a Lagrangian formulation, that compare these choices to likely more realistic boundary condition choices like the analytical solution for flow associated with two diverging plates stretching over a finite-width region. We also compare embedded and nested meshes with a high-resolution 2-D region within a cartesian 'whole mantle cross-section' box. Our initial results imply that the choice of far-field boundary conditions does indeed significantly influence predicted melting distributions and melt volumes associated with continental breakup. For calculations including asthenospheric melting, the 'finite width plate spreading' and embedded rifting boundary condition treatments lead to significantly smaller BC-influenced signals when using high-resolution calculation regions of order ~1000 km wide and 600 km deep within a lower resolution box of the order of >5000 km wide and 2800 km. We recommend their use when models are attempting to resolve the effects of asthenosphere flow and melting. We also discuss several examples of typical numerical 'artifacts' related to 'edge convection' at the sides of the stretching region

  10. A High Order Discontinuous Galerkin Method for 2D Incompressible Flows

    NASA Technical Reports Server (NTRS)

    Liu, Jia-Guo; Shu, Chi-Wang

    1999-01-01

    In this paper we introduce a high order discontinuous Galerkin method for two dimensional incompressible flow in vorticity streamfunction formulation. The momentum equation is treated explicitly, utilizing the efficiency of the discontinuous Galerkin method The streamfunction is obtained by a standard Poisson solver using continuous finite elements. There is a natural matching between these two finite element spaces, since the normal component of the velocity field is continuous across element boundaries. This allows for a correct upwinding gluing in the discontinuous Galerkin framework, while still maintaining total energy conservation with no numerical dissipation and total enstrophy stability The method is suitable for inviscid or high Reynolds number flows. Optimal error estimates are proven and verified by numerical experiments.

  11. Activating Receptor NKG2D Targets RAE-1-Expressing Allogeneic Neural Precursor Cells in a Viral Model of Multiple Sclerosis

    PubMed Central

    Weinger, Jason G.; Plaisted, Warren C.; Maciejewski, Sonia M.; Lanier, Lewis L.; Walsh, Craig M.; Lane, Thomas E.

    2014-01-01

    Transplantation of major histocompatibility complex (MHC)-mismatched mouse neural precursor cells (NPCs) into mice persistently infected with the neurotropic JHM strain of mouse hepatitis virus (JHMV) results in rapid rejection that is mediated, in part, by T cells. However, the contribution of the innate immune response to allograft rejection in a model of viral-induced neurological disease has not been well defined. Herein, we demonstrate that the natural killer (NK) cell-expressing activating receptor NKG2D participates in transplanted allogeneic NPC rejection in mice persistently infected with JHMV. Cultured NPCs derived from C57BL/6 (H-2b) mice express the NKG2D ligand retinoic acid early precursor transcript (RAE)-1 but expression was dramatically reduced upon differentiation into either glia or neurons. RAE-1+ NPCs were susceptible to NK cell-mediated killing whereas RAE-1- cells were resistant to lysis. Transplantation of C57BL/6-derived NPCs into JHMV-infected BALB/c (H-2d) mice resulted in infiltration of NKG2D+CD49b+ NK cells and treatment with blocking antibody specific for NKG2D increased survival of allogeneic NPCs. Further, transplantation of differentiated RAE-1- allogeneic NPCs into JHMV-infected BALB/c mice resulted in enhanced survival, highlighting a role for the NKG2D:RAE-1 signaling axis in allograft rejection. We also demonstrate that transplantation of allogeneic NPCs into JHMV-infected mice resulted in infection of the transplanted cells suggesting that these cells may be targets for infection. Viral infection of cultured cells increased RAE-1 expression, resulting in enhanced NK cell-mediated killing through NKG2D recognition. Collectively, these results show that in a viral-induced demyelination model, NK cells contribute to rejection of allogeneic NPCs through an NKG2D signaling pathway. PMID:24898518

  12. Film flows and self-organized patterns of 2D-localized structures

    SciTech Connect

    Frenkel, A.L.

    1996-12-31

    Films flowing down an inclined plane are considered. An unconventional perturbation approach is discussed. It yields the most general evolution equation for film thickness and the least restrictive conditions for its validity. Results of numerical simulations of the dissipative-dispersive evolution equation indicate that novel, more complex type of spatiotemporal patterns can exist for strange attractors of nonequilibrium systems. It is suggested that real-life experiments satisfying the validity conditions of this theory are possible.

  13. Locally modified QUICK scheme for highly convective 2-D and 3-D flows

    NASA Astrophysics Data System (ADS)

    Leonard, B. P.

    The positive and negative aspects of the QUICK scheme are discussed. QUICK is used in the bulk of the flow domain; however, when the local curvature of the convected variable exceeds a preset value, the algorithm switches to exponential upwinding or other compatible interpolation. Results are presented for the purely convective oblique-step test. A comparison is made between the sharp monotonic EULER-QUICK results and first-, second-, and third-order upwinding.

  14. Extraction of Individual Filaments from 2D Confocal Microscopy Images of Flat Cells.

    PubMed

    Basu, Saurav; Chi Liu; Rohde, Gustavo Kunde

    2015-01-01

    A crucial step in understanding the architecture of cells and tissues from microscopy images, and consequently explain important biological events such as wound healing and cancer metastases, is the complete extraction and enumeration of individual filaments from the cellular cytoskeletal network. Current efforts at quantitative estimation of filament length distribution, architecture and orientation from microscopy images are predominantly limited to visual estimation and indirect experimental inference. Here we demonstrate the application of a new algorithm to reliably estimate centerlines of biological filament bundles and extract individual filaments from the centerlines by systematically disambiguating filament intersections. We utilize a filament enhancement step followed by reverse diffusion based filament localization and an integer programming based set combination to systematically extract accurate filaments automatically from microscopy images. Experiments on simulated and real confocal microscope images of flat cells (2D images) show efficacy of the new method.

  15. A 2-D Implicit, Energy and Charge Conserving Particle In Cell Method

    SciTech Connect

    McPherson, Allen L.; Knoll, Dana A.; Cieren, Emmanuel B.; Feltman, Nicolas; Leibs, Christopher A.; McCarthy, Colleen; Murthy, Karthik S.; Wang, Yijie

    2012-09-10

    Recently, a fully implicit electrostatic 1D charge- and energy-conserving particle-in-cell algorithm was proposed and implemented by Chen et al ([2],[3]). Central to the algorithm is an advanced particle pusher. Particles are moved using an energy conserving scheme and are forced to stop at cell faces to conserve charge. Moreover, a time estimator is used to control errors in momentum. Here we implement and extend this advanced particle pusher to include 2D and electromagnetic fields. Derivations of all modifications made are presented in full. Special consideration is taken to ensure easy coupling into the implicit moment based method proposed by Taitano et al [19]. Focus is then given to optimizing the presented particle pusher on emerging architectures. Two multicore implementations, and one GPU (Graphics Processing Unit) implementation are discussed and analyzed.

  16. Numerical solution of 2D wet steam flow with non-equilibrium condensation and real thermodynamics

    SciTech Connect

    Hric, V.; Halama, J.

    2015-03-10

    An approach to modeling of wet steam flow with non-equilibrium condensation phenomenon is presented. The first part of our flow model is homogeneous Euler system of transport equations for mass, momentum and total energy of wet steam (mixture). The additional second part describes liquid phase via non-homogeneous system of transport equations for moments of droplets number distribution function and relies on corrected classical nucleation theory. Moment equations are closed by linearization of droplet growth rate model. All necessary relations for thermodynamic properties of steam are provided by IAPWS set of equations. However, properties of condensate are simply modeled by liquid saturation data. Two real equations of state are implemented. Recently developed CFD formulation for entropy (does not require iteration process) and so-called IAPWS special gas equation for Helmholtz energy (one iteration loop is necessary). Flow model is validated on converging-diverging supersonic nozzle with Barschdorff geometry. Simulations were performed by in-house CFD code based on finite volume method and stiff character of equations was solved by symmetrical time operator splitting. Achieved results satisfactorily agreed with experimental data.

  17. Enhanced Kalman Filtering for a 2D CFD NS Wind Farm Flow Model

    NASA Astrophysics Data System (ADS)

    Doekemeijer, B. M.; van Wingerden, J. W.; Boersma, S.; Pao, L. Y.

    2016-09-01

    Wind turbines are often grouped together for financial reasons, but due to wake development this usually results in decreased turbine lifetimes and power capture, and thereby an increased levelized cost of energy (LCOE). Wind farm control aims to minimize this cost by operating turbines at their optimal control settings. Most state-of-the-art control algorithms are open-loop and rely on low fidelity, static flow models. Closed-loop control relying on a dynamic model and state observer has real potential to further decrease wind's LCOE, but is often too computationally expensive for practical use. In this paper two time-efficient Kalman filter (KF) variants are outlined incorporating the medium fidelity, dynamic flow model “WindFarmSimulator” (WFSim). This model relies on a discretized set of Navier-Stokes equations in two dimensions to predict the flow in wind farms at low computational cost. The filters implemented are an Ensemble KF and an Approximate KF. Simulations in which a high fidelity simulation model represents the true wind farm show that these filters are 101 —102 times faster than a regular KF with comparable or better performance, correcting for wake dynamics that are not modeled in WFSim (noticeably, wake meandering and turbine hub effects). This is a first big step towards real-time closed-loop control for wind farms.

  18. Symplectically invariant flow equations for N = 2, D = 4 gauged supergravity with hypermultiplets

    NASA Astrophysics Data System (ADS)

    Klemm, Dietmar; Petri, Nicolò; Rabbiosi, Marco

    2016-04-01

    We consider N = 2 supergravity in four dimensions, coupled to an arbitrary number of vector- and hypermultiplets, where abelian isometries of the quaternionic hyperscalar target manifold are gauged. Using a static and spherically or hyperbolically symmetric ansatz for the fields, a one-dimensional effective action is derived whose variation yields all the equations of motion. By imposing a sort of Dirac charge quantization condition, one can express the complete scalar potential in terms of a superpotential and write the action as a sum of squares. This leads to first-order flow equations, that imply the second-order equations of motion. The first-order flow turns out to be driven by Hamilton's characteristic function in the Hamilton-Jacobi formalism, and contains among other contributions the superpotential of the scalars. We then include also magnetic gaugings and generalize the flow equations to a symplectically covariant form. Moreover, by rotating the charges in an appropriate way, an alternative set of non-BPS first-order equations is obtained that corresponds to a different squaring of the action. Finally, we use our results to derive the attractor equations for near-horizon geometries of extremal black holes.

  19. Video lensfree microscopy of 2D and 3D culture of cells

    NASA Astrophysics Data System (ADS)

    Allier, C. P.; Vinjimore Kesavan, S.; Coutard, J.-G.; Cioni, O.; Momey, F.; Navarro, F.; Menneteau, M.; Chalmond, B.; Obeid, P.; Haguet, V.; David-Watine, B.; Dubrulle, N.; Shorte, S.; van der Sanden, B.; Di Natale, C.; Hamard, L.; Wion, D.; Dolega, M. E.; Picollet-D'hahan, N.; Gidrol, X.; Dinten, J.-M.

    2014-03-01

    Innovative imaging methods are continuously developed to investigate the function of biological systems at the microscopic scale. As an alternative to advanced cell microscopy techniques, we are developing lensfree video microscopy that opens new ranges of capabilities, in particular at the mesoscopic level. Lensfree video microscopy allows the observation of a cell culture in an incubator over a very large field of view (24 mm2) for extended periods of time. As a result, a large set of comprehensive data can be gathered with strong statistics, both in space and time. Video lensfree microscopy can capture images of cells cultured in various physical environments. We emphasize on two different case studies: the quantitative analysis of the spontaneous network formation of HUVEC endothelial cells, and by coupling lensfree microscopy with 3D cell culture in the study of epithelial tissue morphogenesis. In summary, we demonstrate that lensfree video microscopy is a powerful tool to conduct cell assays in 2D and 3D culture experiments. The applications are in the realms of fundamental biology, tissue regeneration, drug development and toxicology studies.

  20. NKG2D functions as an activating receptor on natural killer cells in the common marmoset (Callithrix jacchus).

    PubMed

    Watanabe, Masamichi; Kudo, Yohei; Kawano, Mitsuko; Nakayama, Masafumi; Nakamura, Kyohei; Kameda, Mai; Ebara, Masamune; Sato, Takeki; Nakamura, Marina; Omine, Kaito; Kametani, Yoshie; Suzuki, Ryuji; Ogasawara, Kouetsu

    2014-11-01

    The natural killer group 2 membrane D (NKG2D) receptor is an NK-activating receptor that plays an important role in host defense against tumors and viral infections. Although the marmoset is an important and reliable animal model, especially for the study of human-specific viral infections, functional characterization of NKG2D on marmoset NK cells has not previously been conducted. In the present study, we investigated a subpopulation of marmoset NK cells that express NKG2D and exhibit cytolytic potential. On the basis of their CD16 and CD56 expression patterns, marmoset NK cells can be classified into three subpopulations: CD16(+) CD56(-), CD16(-) CD56(+) and CD16(-) CD56(-) cells. NKG2D expression on marmoset CD16(+) CD56(-) and CD16(-) CD56(+) splenocytes was confirmed using an NKG2D ligand composed of an MHC class I chain-related molecule A (MICA)-Fc fusion protein. When marmoset splenocytes were cultured with IL-2 for 4 days, NKG2D expression was retained on CD16(+) CD56(-) and CD16(-) CD56(+). In addition, CD16(+) CD56(+) cells within the marmoset NK population appeared which expressed NKG2D after IL-2 stimulation. IL-2-activated marmoset NK cells showed strong cytolytic activity against K562 target cells and target cells stably expressing MICA. Further, the cytolytic activity of marmoset splenocytes was significantly reduced after addition of MICA-Fc fusion protein. Thus, NKG2D functions as an activating receptor on marmoset NK cells that possesses cytotoxic potential, and phenotypic profiles of marmoset NK cell subpopulations are similar to those seen in humans. PMID:24860119

  1. Myocyte enhancer factor 2D regulates ectoderm specification and adhesion properties of animal cap cells in the early Xenopus embryo.

    PubMed

    Katz Imberman, Sandra; Kolpakova, Alina; Keren, Aviad; Bengal, Eyal

    2015-08-01

    In Xenopus, animal cap (AC) cells give rise to ectoderm and its derivatives: epidermis and the central nervous system. Ectoderm has long been considered a default pathway of embryonic development, with cells that are not under the influence of vegetal Nodal signaling adopting an ectodermal program of gene expression. In the present study, we describe the involvement of the animally-localized maternal transcription factor myocyte enhancer factor (Mef) 2D in regulating the identity of AC cells. We find that Mef2D is required for the formation of both ectodermal lineages: neural and epidermis. Gain and loss of function experiments indicate that Mef2D regulates early gastrula expression of key ectodermal/epidermal genes in the animal region. Mef2D controls the activity of zygotic bone morphogenetic protein (BMP) signaling known to dictate the epidermal differentiation program. Exogenous expression of Mef2D in vegetal blastomeres was sufficient to induce ectopic expression of ectoderm/epidermal genes in the vegetal half of the embryo, when Nodal signaling was inhibited. Depletion of Mef2D caused a loss of AC cell adhesion that was rescued by the expression of E-cadherin or bone morphogenetic protein 4. In addition, expression of Mef2D in the prospective endoderm caused unusual aggregation of vegetal cells with animal cells in vitro and inappropriate segregation to other germ layers in vivo. Mef2D cooperates with another animally-expressed transcription factor, FoxI1e. Together, they regulate the expression of genes encoding signaling proteins and the transcription factors that control the regional identity of animal cells. Therefore, we describe a new role for the animally-localized Mef2D protein in early ectoderm specification, which is similar to that of the vegetally-localized VegT in endoderm and mesoderm formation.

  2. The histone lysine methyltransferase KMT2D sustains a gene expression program that represses B cell lymphoma development.

    PubMed

    Ortega-Molina, Ana; Boss, Isaac W; Canela, Andres; Pan, Heng; Jiang, Yanwen; Zhao, Chunying; Jiang, Man; Hu, Deqing; Agirre, Xabier; Niesvizky, Itamar; Lee, Ji-Eun; Chen, Hua-Tang; Ennishi, Daisuke; Scott, David W; Mottok, Anja; Hother, Christoffer; Liu, Shichong; Cao, Xing-Jun; Tam, Wayne; Shaknovich, Rita; Garcia, Benjamin A; Gascoyne, Randy D; Ge, Kai; Shilatifard, Ali; Elemento, Olivier; Nussenzweig, Andre; Melnick, Ari M; Wendel, Hans-Guido

    2015-10-01

    The gene encoding the lysine-specific histone methyltransferase KMT2D has emerged as one of the most frequently mutated genes in follicular lymphoma and diffuse large B cell lymphoma; however, the biological consequences of KMT2D mutations on lymphoma development are not known. Here we show that KMT2D functions as a bona fide tumor suppressor and that its genetic ablation in B cells promotes lymphoma development in mice. KMT2D deficiency also delays germinal center involution and impedes B cell differentiation and class switch recombination. Integrative genomic analyses indicate that KMT2D affects methylation of lysine 4 on histone H3 (H3K4) and expression of a set of genes, including those in the CD40, JAK-STAT, Toll-like receptor and B cell receptor signaling pathways. Notably, other KMT2D target genes include frequently mutated tumor suppressor genes such as TNFAIP3, SOCS3 and TNFRSF14. Therefore, KMT2D mutations may promote malignant outgrowth by perturbing the expression of tumor suppressor genes that control B cell-activating pathways. PMID:26366710

  3. The histone lysine methyltransferase KMT2D sustains a gene expression program that represses B cell lymphoma development

    PubMed Central

    Ortega-Molina, Ana; Boss, Isaac W.; Canela, Andres; Pan, Heng; Jiang, Yanwen; Zhao, Chunying; Jiang, Man; Hu, Deqing; Agirre, Xabier; Niesvizky, Itamar; Lee, Ji-Eun; Chen, Hua-Tang; Ennishi, Daisuke; Scott, David W.; Mottok, Anja; Hother, Christoffer; Liu, Shichong; Cao, Xing-Jun; Tam, Wayne; Shaknovich, Rita; Garcia, Benjamin A.; Gascoyne, Randy D.; Ge, Kai; Shilatifard, Ali; Elemento, Olivier; Nussenzweig, Andre; Melnick, Ari M.; Wendel, Hans-Guido

    2015-01-01

    The lysine-specific histone methyltransferase KMT2D has emerged as one of the most frequently mutated genes in follicular lymphoma (FL) and diffuse large B cell lymphoma (DLBCL). However, the biological consequences of KMT2D mutations on lymphoma development are not known. Here we show that KMT2D functions as a bona fide tumor suppressor and that its genetic ablation in B cells promotes lymphoma development in mice. KMT2D deficiency also delays germinal center (GC) involution, impedes B cell differentiation and class switch recombination (CSR). Integrative genomic analyses indicate that KMT2D affects H3K4 methylation and expression of a specific set of genes including those in the CD40, JAK-STAT, Toll-like receptor, and B cell receptor pathways. Notably, other KMT2D target genes include frequently mutated tumor suppressor genes such as TNFAIP3, SOCS3, and TNFRSF14. Therefore, KMT2D mutations may promote malignant outgrowth by perturbing the expression of tumor suppressor genes that control B cell activating pathways. PMID:26366710

  4. Analysis of High-Speed Rotating Flow in 2D Polar (r - θ)Coordinate

    NASA Astrophysics Data System (ADS)

    Pradhan, S.

    2016-03-01

    The generalized analytical model for the radial boundary layer in a high-speed rotating cylinder is formulated for studying the gas flow field due to insertion of mass, momentum and energy into the rotating cylinder in the polar (r - θ) plane. The analytical solution includes the sixth order differential equation for the radial boundary layer at the cylindrical curved surface in terms of master potential (χ) , which is derived from the equations of motion in a polar (r - θ) plane. The linearization approximation (Wood & Morton, J. Fluid Mech-1980; Pradhan & Kumaran, J. Fluid Mech-2011; Kumaran & Pradhan, J. Fluid Mech-2014) is used, where the equations of motion are truncated at linear order in the velocity and pressure disturbances to the base flow, which is a solid-body rotation. Additional assumptions in the analytical model include constant temperature in the base state (isothermal condition), and high Reynolds number, but there is no limitation on the stratification parameter. In this limit, the gas flow is restricted to a boundary layer of thickness (Re (1 / 3) R) at the wall of the cylinder. Here, the stratification parameter A = √ ((mΩ 2R2) / (2kB T)) . This parameter Ais the ratio of the peripheral speed, ΩR , to the most probable molecular speed, √(2 k_B T/m), the Reynolds number Re = (ρ _w ΩR2 / μ) , where m is the molecular mass, Ω and R are the rotational speed and radius of the cylinder, k_B is the Boltzmann constant, T is the gas temperature, ρ_w is the gas density at wall, and μ is the gas viscosity. The analytical solutions are then compared with direct simulation Monte Carlo (DSMC) simulations.

  5. Development of 2D dynamic model for hydrogen-fed and methane-fed solid oxide fuel cells

    NASA Astrophysics Data System (ADS)

    Luo, X. J.; Fong, K. F.

    2016-10-01

    A new two-dimensional (2D) dynamic model is developed in Fortran to study the mass and energy transport, the velocity field and the electrochemical phenomena of high-temperature solid oxide fuel cell (SOFC). The key feature of this model is that gas properties, reaction heat, open circuit voltage, ohmic voltage and exchange current density are temperature-dependent. Based on this, the change of gas temperature and related characteristics can be evaluated in this study. The transient performances of SOFC, like heat-up and start-up processes, are therefore assessed accordingly. In this 2D dynamic SOFC model, chemical and electrochemical reaction, flow field, mass and energy transfer models are coupled in order to determine the current density, the mass fraction and the temperature of gas species. Mass, momentum and energy balance equations are discretized by finite difference method. Performance evaluation in current density, electrical efficiency and overall efficiency is conducted for the effects of different operating parameters in SOFC. The present model can serve as a valuable tool for in-depth performance evaluation of other design and operating parameters of SOFC unit, as well as further dynamic simulation and optimization of SOFC as a prime mover in cogeneration or trigeneration system.

  6. SWAT and River-2D Modelling of Pinder River for Analysing Snow Trout Habitat under Different Flow Abstraction Scenarios

    NASA Astrophysics Data System (ADS)

    Nale, J. P.; Gosain, A. K.; Khosa, R.

    2015-12-01

    Pinder River, one of major headstreams of River Ganga, originates in Pindari Glaciers of Kumaon Himalayas and after passing through rugged gorges meets Alaknanda at Karanprayag forming one of the five celestial confluences of Upper Ganga region. While other sub-basins of Upper Ganga are facing severe ecological losses, Pinder basin is still in its virginal state and is well known for its beautiful valleys besides being host to unique and rare biodiversity. A proposed 252 MW run-of-river hydroelectric project at Devsari on this river has been a major concern on account of its perceived potential for egregious environmental and social impacts. In this context, the study presented tries to analyse the expected changes in aquatic habitat conditions after this project is operational (with different operation policies). SWAT hydrological modelling platform has been used to derive stream flow simulations under various scenarios ranging from the present to the likely future conditions. To analyse the habitat conditions, a two dimensional hydraulic-habitat model 'River-2D', a module of iRIC software, is used. Snow trout has been identified as the target keystone species and its habitat preferences, in the form of flow depths, flow velocity and substrate condition, are obtained from diverse sources of related literature and are provided as Habitat Suitability Indices to River-2D. Bed morphology constitutes an important River-2D input and has been obtained, for the designated 1 km long study reach of Pinder upto Karanprayag, from a combination of actual field observations and supplemented by SRTM 1 Arc-Second Global digital elevation data. Monthly Weighted Usable Area for three different life stages (Spawning, Juvenile and Adult) of Snow Trout are obtained corresponding to seven different flow discharges ranging from 10 cumec to 1000 cumec. Comparing the present and proposed future river flow conditions obtained from SWAT modelling, losses in Weighted Usable Area, for the

  7. Emergent Criticality and Ricci Flow in a 2D Frustrated Heisenberg Model

    NASA Astrophysics Data System (ADS)

    Orth, Peter P.

    2014-03-01

    In most systems that exhibit order at low temperatures, the order occurs in the elementary degrees of freedom such as spin or charge. Prominent examples are magnetic or superconducting states of matter. In contrast, emergent order describes the phenomenon where composite objects exhibit longer range correlations. Such emergent order has been suspected to occur in a range of correlated materials. One specific example are spin systems with competing interactions, where long-range discrete order in the relative orientation of spins may occur. Interestingly, this order parameter may induce other phase transitions as is the case for the nematic transition in the iron pnictides. In this talk, we introduce and discuss a system with emergent Z6 symmetry, a two-dimensional frustrated Heisenberg antiferromagnet on the windmill lattice consisting of interpenetrating honeycomb and triangular lattices. The multiple spin stiffnesses can be captured in terms of a four-dimensional metric tensor, and the renormalization group flow of the stiffnesses is described by the Ricci flow of the metric tensor. The key result is a decoupling of an emergent collective degree of freedom given by the relative phase of spins on different sublattices. In particular, our results reveal a sequence of two Berezinskii-Kosterlitz-Thouless phase transitions that bracket a critical phase.

  8. Observing rearrangements in a 2D emulsion flowing through a hopper

    NASA Astrophysics Data System (ADS)

    Chen, Dandan; Desmond, Ken; Weeks, Eric R.

    2010-03-01

    Jamming in granular flow through a hopper has been well studied, and structures such as arches have been found in simulations both with and without friction, and in experiments with friction. To study if jamming can happen in other frictionless systems, we pump dense emulsions (oil in water) through a glass hopper. The oil droplets experience a viscous friction but do not have static friction acting between touching droplets, in contrast to granular particles. For easy imaging, we squeeze the droplets into quasi two-dimensional disks by injecting the emulsion into a thin chamber made from two parallel glass plates. Movies of the flow are taken from the top by a microscope. Due to the narrowing confinement in the hopper, droplets are forced to rearrange, and we observe topological changes such as T1 events. At the same time, the interdroplet forces are measured from the deformation of the droplets. By varying the hopper gap width and angle, we study how the constriction affects the particles' motions, and how this relates to the interdroplet forces.

  9. Manifest: A computer program for 2-D flow modeling in Stirling machines

    NASA Technical Reports Server (NTRS)

    Gedeon, David

    1989-01-01

    A computer program named Manifest is discussed. Manifest is a program one might want to use to model the fluid dynamics in the manifolds commonly found between the heat exchangers and regenerators of Stirling machines; but not just in the manifolds - in the regenerators as well. And in all sorts of other places too, such as: in heaters or coolers, or perhaps even in cylinder spaces. There are probably nonStirling uses for Manifest also. In broad strokes, Manifest will: (1) model oscillating internal compressible laminar fluid flow in a wide range of two-dimensional regions, either filled with porous materials or empty; (2) present a graphics-based user-friendly interface, allowing easy selection and modification of region shape and boundary condition specification; (3) run on a personal computer, or optionally (in the case of its number-crunching module) on a supercomputer; and (4) allow interactive examination of the solution output so the user can view vector plots of flow velocity, contour plots of pressure and temperature at various locations and tabulate energy-related integrals of interest.

  10. Natural killer cell NKG2D and granzyme B are critical for allergic pulmonary inflammation⋆

    PubMed Central

    Farhadi, Nazanin; Lambert, Laura; Triulzi, Chiara; Openshaw, Peter J.M.; Guerra, Nadia; Culley, Fiona J.

    2014-01-01

    Background The diverse roles of innate immune cells in the pathogenesis of asthma remain to be fully defined. Natural killer (NK) cells are innate lymphocytes that can regulate adaptive immune responses. NK cells are activated in asthma; however, their role in allergic airway inflammation is not fully understood. Objective We investigated the importance of NK cells in house dust mite (HDM)-triggered allergic pulmonary inflammation. Specifically, we aimed to determine the role of the major NK-cell activating receptor NKG2D and NK-cell effector functions mediated by granzyme B. Methods Allergic airway inflammation was induced in the airways of mice by repeated intranasal HDM extract administration and responses in wild-type and NKG2D-deficient mice were compared. Adoptive transfer studies were used to identify the cells and mechanisms involved. Results Mice that lacked NKG2D were resistant to the induction of allergic inflammation and showed little pulmonary eosinophilia, few airway TH2 cells, and no rise in serum IgE after multiple HDM-allergen exposures. However, NKG2D was not required for pulmonary inflammation after a single inoculation of allergen. NKG2D-deficient mice showed no alteration in responses to respiratory virus infection. Transfer of wild-type NK cells (but not CD3+ cells) into NKG2D-deficient mice restored allergic inflammatory responses only if the NK cells expressed granzyme B. Conclusions These studies established a pivotal role for NK-cell NKG2D and granzyme B in the pathogenesis of HDM-induced allergic lung disease, and identified novel therapeutic targets for the prevention and treatment of asthma. PMID:24290277

  11. A Beam-Fourier Technique for the Numerical Investigation of 2D Nonlinear Convective Flows

    NASA Astrophysics Data System (ADS)

    Papanicolaou, N. C.

    2011-11-01

    In the current work, we develop a numerical method suitable for treating the problem of nonlinear two-dimensional flows in rectangular domains. For the spatial approximation we employ the Fourier-Galerkin approach. More specifically, our basis functions are products of trigonometric and Beam functions. This choice means that the solutions automatically satisfy the boundary and periodic conditions in the x and y directions respectively. The accuracy of the method is assessed by applying it to model problems which admit exact analytical solutions. The numerical and analytic solutions are found to be in good agreement. The convergence rate of the spectral coefficients is found to be fifth-order algebraic in the x-direction and y-direction, confirming the efficiency and speed of our technique.

  12. 2D transient granular flows over obstacles: experimental and numerical work

    NASA Astrophysics Data System (ADS)

    Juez, Carmelo; Caviedes-Voullième, Daniel; Murillo, Javier; García-Navarro, Pilar

    2016-04-01

    Landslides are an ubiquitous natural hazard, and therefore human infrastructure and settlements are often at risk in mountainous regions. In order to better understand and predict landslides, systematic studies of the phenomena need to be undertaken. In particular, computational tools which allow for analysis of field problems require to be thoroughly tested, calibrated and validated under controlled conditions. And to do so, it is necessary for such controlled experiments to be fully characterized in the same terms as the numerical model requires. This work presents an experimental study of dry granular flow over a rough bed with topography which resembles a mountain valley. It has an upper region with a very high slope. The geometry of the bed describes a fourth order polynomial curve, with a low point with zero slope, and afterwards a short region with adverse slope. Obstacles are present in the lower regions which are used as model geometries of human structures. The experiments consisted of a sudden release a mass of sand on the upper region, and allowing it to flow downslope. Furthermore, it has been frequent in previous studies to measure final states of the granular mass at rest, but seldom has transient data being provided, and never for the entire field. In this work we present transient measurements of the moving granular surfaces, obtained with a consumer-grade RGB-D sensor. The sensor, developed for the videogame industry, allows to measure the moving surface of the sand, thus obtaining elevation fields. The experimental results are very consistent and repeatable. The measured surfaces clearly show the distinctive features of the granular flow around the obstacles and allow to qualitatively describe the different flow patterns. More importantly, the quantitative description of the granular surface allows for benchmarking and calibration of predictive numerical models, key in scaling the small-scale experimental knowledge into the field. In addition, as

  13. Modeling water flow and nitrate dynamics in a plastic mulch vegetable cultivation system using HYDRUS-2D

    NASA Astrophysics Data System (ADS)

    Filipović, Vilim; Romić, Davor; Romić, Marija; Matijević, Lana; Mallmann, Fábio J. K.; Robinson, David A.

    2016-04-01

    Growing vegetables commercially requires intensive management and involves high irrigation demands and input of agrochemicals. Plastic mulch application in combination with drip irrigation is a common agricultural management technique practiced due to variety of benefits to the crop, mostly vegetable biomass production. However, the use of these techniques can result in various impacts on water and nutrient distribution in underlying soil and consequently affect nutrient leaching towards groundwater resources. The aim of this work is to estimate the effect of plastic mulch cover in combination with drip irrigation on water and nitrate dynamics in soil using HYDRUS-2D model. The field site was located in Croatian costal karst area on a Gleysol (WRB). The experiment was designed according to the split-plot design in three repetitions and was divided into plots with plastic mulch cover (MULCH) and control plots with bare soil (CONT). Each of these plots received applications of three levels of nitrogen fertilizer: 70, 140, and 210 kg per ha. All plots were equipped with drip irrigation and cropped with bell pepper (Capsicum annuum L. cv. Bianca F1). Lysimeters were installed at 90 cm depth in all plots and were used for monitoring the water and nitrate outflow. HYDRUS-2D was used for modeling the water and nitrogen outflow in the MULCH and CONT plots, implementing the proper boundary conditions. HYDRUS-2D simulated results showed good fitting to the field site observed data in both cumulative water and nitrate outflow, with high level of agreement. Water flow simulations produced model efficiency of 0.84 for CONT and 0.56 for MULCH plots, while nitrate simulations showed model efficiency ranging from 0.67 to 0.83 and from 0.70 to 0.93, respectively. Additional simulations were performed with the absence of the lysimeter, revealing faster transport of nitrates below drip line in the CONT plots, mostly because of the increased surface area subjected to precipitation

  14. Lensfree video microscopy: high throughput monitoring and cell tracking of 2D cell cultures

    NASA Astrophysics Data System (ADS)

    Allier, C. P.; Vinjimore Kesavan, S.; Cioni, O.; Momey, F.; Bordy, T.; Hervé, L.; Morel, S.; Navarro, F.; Menneteau, M.; Chalmond, B.; Freida, D.; Sulpice, E.; Gidrol, X.; Dinten, J.-M.

    2015-07-01

    In order to extend the analysis of the datasets produced by lensfree video microscopy we have implemented a cell tracking algorithm to combine and correlate cell motility to the previously devised metrics to quantify e.g. cell adhesion and spreading, cell division, and cell death. In this paper we present the assessment of these new methodology on experiments involving three different cell lines, namely 3T3 fibroblast cells, primary HUVEC cells and macrophage THP1 cells. We demonstrate that the good spatial resolution and the fast frame rate obtained with of our lensfree video microscope allows standard cell tracking algorithm to be computed. The results is the possibility to analyze thousands of cells successfully tracked over tens of hours. The results is the possibility to compare different cell cultures in terms of e.g. cell motility and cell confinement ration. Ultimately we managed to measure the doubling time at single cell level over a large number of N=235 cells tracked over two days.

  15. Presentation of the MERC work-flow for the computation of a 2D radial reflector in a PWR

    SciTech Connect

    Clerc, T.; Hebert, A.; Leroyer, H.; Argaud, J. P.; Poncot, A.; Bouriquet, B.

    2013-07-01

    This paper presents a work-flow for computing an equivalent 2D radial reflector in a pressurized water reactor (PWR) core, in adequacy with a reference power distribution, computed with the method of characteristics (MOC) of the lattice code APOLLO2. The Multi-modelling Equivalent Reflector Computation (MERC) work-flow is a coherent association of the lattice code APOLLO2 and the core code COCAGNE, structured around the ADAO (Assimilation de Donnees et Aide a l'Optimisation) module of the SALOME platform, based on the data assimilation theory. This study leads to the computation of equivalent few-groups reflectors, that can be spatially heterogeneous, which have been compared to those obtained with the OPTEX similar methodology developed with the core code DONJON, as a first validation step. Subsequently, the MERC work-flow is used to compute the most accurate reflector in consistency with all the R and D choices made at Electricite de France (EDF) for the core modelling, in terms of number of energy groups and simplified transport solvers. We observe important reductions of the power discrepancies distribution over the core when using equivalent reflectors obtained with the MERC work-flow. (authors)

  16. Common Effects on Cancer Cells Exerted by a Random Positioning Machine and a 2D Clinostat.

    PubMed

    Svejgaard, Benjamin; Wehland, Markus; Ma, Xiao; Kopp, Sascha; Sahana, Jayashree; Warnke, Elisabeth; Aleshcheva, Ganna; Hemmersbach, Ruth; Hauslage, Jens; Grosse, Jirka; Bauer, Johann; Corydon, Thomas Juhl; Islam, Tawhidul; Infanger, Manfred; Grimm, Daniela

    2015-01-01

    In this study we focused on gravity-sensitive proteins of two human thyroid cancer cell lines (ML-1; RO82-W-1), which were exposed to a 2D clinostat (CLINO), a random positioning machine (RPM) and to normal 1g-conditions. After a three (3d)- or seven-day-culture (7d) on the two devices, we found both cell types growing three-dimensionally within multicellular spheroids (MCS) and also cells remaining adherent (AD) to the culture flask, while 1g-control cultures only formed adherent monolayers, unless the bottom of the culture dish was covered by agarose. In this case, the cytokines IL-6 and IL-8 facilitated the formation of MCS in both cell lines using the liquid-overlay technique at 1g. ML-1 cells grown on the RPM or the CLINO released amounts of IL-6 and MCP-1 into the supernatant, which were significantly elevated as compared to 1g-controls. Release of IL-4, IL-7, IL-8, IL-17, eotaxin-1 and VEGF increased time-dependently, but was not significantly influenced by the gravity conditions. After 3d on the RPM or the CLINO, an accumulation of F-actin around the cellular membrane was detectable in AD cells of both cell lines. IL-6 and IL-8 stimulation of ML-1 cells for 3d and 7d influenced the protein contents of ß1-integrin, talin-1, Ki-67, and beta-actin dose-dependently in adherent cells. The ß1-integrin content was significantly decreased in AD and MCS samples compared with 1g, while talin-1 was higher expressed in MCS than AD populations. The proliferation marker Ki-67 was elevated in AD samples compared with 1g and MCS samples. The ß-actin content of R082-W-1 cells remained unchanged. ML-1 cells exhibited no change in ß-actin in RPM cultures, but a reduction in CLINO samples. Thus, we concluded that simulated microgravity influences the release of cytokines in follicular thyroid cancer cells, and the production of ß1-integrin and talin-1 and predicts an identical effect under real microgravity conditions.

  17. Common Effects on Cancer Cells Exerted by a Random Positioning Machine and a 2D Clinostat

    PubMed Central

    Svejgaard, Benjamin; Wehland, Markus; Ma, Xiao; Kopp, Sascha; Sahana, Jayashree; Warnke, Elisabeth; Aleshcheva, Ganna; Hemmersbach, Ruth; Hauslage, Jens; Grosse, Jirka; Bauer, Johann; Corydon, Thomas Juhl; Islam, Tawhidul; Infanger, Manfred; Grimm, Daniela

    2015-01-01

    In this study we focused on gravity-sensitive proteins of two human thyroid cancer cell lines (ML-1; RO82-W-1), which were exposed to a 2D clinostat (CLINO), a random positioning machine (RPM) and to normal 1g-conditions. After a three (3d)- or seven-day-culture (7d) on the two devices, we found both cell types growing three-dimensionally within multicellular spheroids (MCS) and also cells remaining adherent (AD) to the culture flask, while 1g-control cultures only formed adherent monolayers, unless the bottom of the culture dish was covered by agarose. In this case, the cytokines IL-6 and IL-8 facilitated the formation of MCS in both cell lines using the liquid-overlay technique at 1g. ML-1 cells grown on the RPM or the CLINO released amounts of IL-6 and MCP-1 into the supernatant, which were significantly elevated as compared to 1g-controls. Release of IL-4, IL-7, IL-8, IL-17, eotaxin-1 and VEGF increased time-dependently, but was not significantly influenced by the gravity conditions. After 3d on the RPM or the CLINO, an accumulation of F-actin around the cellular membrane was detectable in AD cells of both cell lines. IL-6 and IL-8 stimulation of ML-1 cells for 3d and 7d influenced the protein contents of ß1-integrin, talin-1, Ki-67, and beta-actin dose-dependently in adherent cells. The ß1-integrin content was significantly decreased in AD and MCS samples compared with 1g, while talin-1 was higher expressed in MCS than AD populations. The proliferation marker Ki-67 was elevated in AD samples compared with 1g and MCS samples. The ß-actin content of R082-W-1 cells remained unchanged. ML-1 cells exhibited no change in ß-actin in RPM cultures, but a reduction in CLINO samples. Thus, we concluded that simulated microgravity influences the release of cytokines in follicular thyroid cancer cells, and the production of ß1-integrin and talin-1 and predicts an identical effect under real microgravity conditions. PMID:26274317

  18. Common Effects on Cancer Cells Exerted by a Random Positioning Machine and a 2D Clinostat.

    PubMed

    Svejgaard, Benjamin; Wehland, Markus; Ma, Xiao; Kopp, Sascha; Sahana, Jayashree; Warnke, Elisabeth; Aleshcheva, Ganna; Hemmersbach, Ruth; Hauslage, Jens; Grosse, Jirka; Bauer, Johann; Corydon, Thomas Juhl; Islam, Tawhidul; Infanger, Manfred; Grimm, Daniela

    2015-01-01

    In this study we focused on gravity-sensitive proteins of two human thyroid cancer cell lines (ML-1; RO82-W-1), which were exposed to a 2D clinostat (CLINO), a random positioning machine (RPM) and to normal 1g-conditions. After a three (3d)- or seven-day-culture (7d) on the two devices, we found both cell types growing three-dimensionally within multicellular spheroids (MCS) and also cells remaining adherent (AD) to the culture flask, while 1g-control cultures only formed adherent monolayers, unless the bottom of the culture dish was covered by agarose. In this case, the cytokines IL-6 and IL-8 facilitated the formation of MCS in both cell lines using the liquid-overlay technique at 1g. ML-1 cells grown on the RPM or the CLINO released amounts of IL-6 and MCP-1 into the supernatant, which were significantly elevated as compared to 1g-controls. Release of IL-4, IL-7, IL-8, IL-17, eotaxin-1 and VEGF increased time-dependently, but was not significantly influenced by the gravity conditions. After 3d on the RPM or the CLINO, an accumulation of F-actin around the cellular membrane was detectable in AD cells of both cell lines. IL-6 and IL-8 stimulation of ML-1 cells for 3d and 7d influenced the protein contents of ß1-integrin, talin-1, Ki-67, and beta-actin dose-dependently in adherent cells. The ß1-integrin content was significantly decreased in AD and MCS samples compared with 1g, while talin-1 was higher expressed in MCS than AD populations. The proliferation marker Ki-67 was elevated in AD samples compared with 1g and MCS samples. The ß-actin content of R082-W-1 cells remained unchanged. ML-1 cells exhibited no change in ß-actin in RPM cultures, but a reduction in CLINO samples. Thus, we concluded that simulated microgravity influences the release of cytokines in follicular thyroid cancer cells, and the production of ß1-integrin and talin-1 and predicts an identical effect under real microgravity conditions. PMID:26274317

  19. Expansion of NK Cells and Reduction of NKG2D Expression in Chronic Lymphocytic Leukemia. Correlation with Progressive Disease

    PubMed Central

    Huergo-Zapico, Leticia; Acebes-Huerta, Andrea; Gonzalez-Rodriguez, Ana Pilar; Contesti, Juan; Gonzalez-García, Esther; Payer, Angel R.; Villa-Alvarez, Monica; Fernández-Guizán, Azahara; López-Soto, Alejandro; Gonzalez, Segundo

    2014-01-01

    The immune system may mediate anti-tumor responses in chronic lymphocytic leukemia (CLL) which may affect disease progression and survival. In this study, we analyzed the immune characteristics of 99 consecutive previously diagnosed CLL patients and 50 healthy controls. The distribution of lymphocyte subsets at diagnosis was retrospectively analyzed. Compared with controls, leukemia patients showed an expansion of NK and CD8 T cells at diagnosis. The relative number of CD8 T cells at diagnosis was associated with time to treatment, suggesting that CD8 T cells may modify disease progression. The distribution of lymphocyte subsets was analyzed again when patients were enrolled in this study. The median time since these patients were diagnosed was 277 weeks. Compared with diagnosis, the absolute number of CD8 T cells significantly decreased in these patients, reaching similar values to healthy controls; however NK cells kept significantly elevated overtime. Nevertheless, NK cells showed an impaired expression of NKG2D receptor and a defective cytotoxic activity. This down-regulation of NKG2D expression was further enhanced in patients with advanced and progressive disease. Additionally, membrane NKG2D levels significantly decreased on CD8 T cells, but a significant increase of NKG2D+CD4+ T cells was observed in CLL patients. The cytotoxic activity of NK cells was diminished in CLL patients; however the treatments with IL-2, IL-15, IL-21 and lenalidomide were able to restore their activity. The effect of IL-2 and IL-15 was associated with the increase of NKG2D expression on immune cells, but the effect of IL-21 and lenalidomide was not due to NKG2D up-regulation. The expansion of NK cells and the reversibility of NK cell defects provide new opportunities for the immunotherapeutic intervention in CLL. PMID:25286418

  20. 2D fluid model analysis for the effect of 3D gas flow on a capacitively coupled plasma deposition reactor

    NASA Astrophysics Data System (ADS)

    Kim, Ho Jun; Lee, Hae June

    2016-06-01

    The wide applicability of capacitively coupled plasma (CCP) deposition has increased the interest in developing comprehensive numerical models, but CCP imposes a tremendous computational cost when conducting a transient analysis in a three-dimensional (3D) model which reflects the real geometry of reactors. In particular, the detailed flow features of reactive gases induced by 3D geometric effects need to be considered for the precise calculation of radical distribution of reactive species. Thus, an alternative inclusive method for the numerical simulation of CCP deposition is proposed to simulate a two-dimensional (2D) CCP model based on the 3D gas flow results by simulating flow, temperature, and species fields in a 3D space at first without calculating the plasma chemistry. A numerical study of a cylindrical showerhead-electrode CCP reactor was conducted for particular cases of SiH4/NH3/N2/He gas mixture to deposit a hydrogenated silicon nitride (SiN x H y ) film. The proposed methodology produces numerical results for a 300 mm wafer deposition reactor which agree very well with the deposition rate profile measured experimentally along the wafer radius.

  1. IL-2-activated haploidentical NK cells restore NKG2D-mediated NK-cell cytotoxicity in neuroblastoma patients by scavenging of plasma MICA.

    PubMed

    Kloess, Stephan; Huenecke, Sabine; Piechulek, Daniel; Esser, Ruth; Koch, Joachim; Brehm, Claudia; Soerensen, Jan; Gardlowski, Tanja; Brinkmann, Andrea; Bader, Peter; Passweg, Jakob; Klingebiel, Thomas; Schwabe, Dirk; Koehl, Ulrike

    2010-11-01

    NK group 2D (NKG2D)-expressing NK cells exhibit cytolytic activity against various tumors after recognition of the cellular ligand MHC class I chain-related gene A (MICA). However, release of soluble MICA (sMICA) compromises NKG2D-dependent NK-cell cytotoxicity leading to tumor escape from immunosurveillance. Although some molecular details of the NKG2D-MICA interaction have been elucidated, its impact for donor NK (dNK) cell-based therapy of solid tumors has not been studied. Within an ongoing phase I/II trial, we used allogeneic IL-2 activated dNK cells after haploidentical stem cell transplantation for immunotherapy of patients with high-risk stage IV neuroblastoma. NKG2D levels on activated dNK cells increased strongly when compared with freshly isolated dNK cells and correlated with enhanced NK-cell cytotoxicity. Most importantly, elevated sMICA levels in patients plasma correlated significantly with impaired dNK-cell-mediated cytotoxicity. This effect could be reversed by high-dose infusion of activated dNK cells, which display high levels of surface NKG2D. Our data suggest that the provided excess of NKG2D leads to clearance of sMICA and preserves cytotoxicity of dNK cells via non-occupied NKG2D. In conclusion, our results identify this tumor immune escape mechanism as a target to improve immunotherapy of neuroblastoma and presumably other tumors.

  2. Drug-induced hyperploidy stimulates an antitumor NK cell response mediated by NKG2D and DNAM-1 receptors

    PubMed Central

    Acebes-Huerta, Andrea; Lorenzo-Herrero, Seila; Folgueras, Alicia R.; Huergo-Zapico, Leticia; Lopez-Larrea, Carlos; López-Soto, Alejandro; Gonzalez, Segundo

    2016-01-01

    ABSTRACT Formation of polyploid or aneuploid cells is a pathological hallmark of malignant tumors. Cell cycle checkpoint mechanisms play a crucial role in ensuring genomic integrity during mitosis, avoiding the generation of aneuploid cells. Additionally, cancer cell DNA ploidy is subjected to extrinsic controls operated by activation of adaptive immune responses mediated by T cells. NK cells exert a central role in the innate anticancer immunity; however, the mechanisms involved in the recognition of tumor cells by NK cells have not been fully elucidated. Herein, we report that drug-induced polyploidy in cancer cells activates antitumor responses mediated by NK cells. Thus, hyperploidy-inducing chemotherapeutic agents strongly upregulate the tumor expression of ligands for the NK cell activating receptors NKG2D and DNAM-1. Drug-induced hyperploidy modulated the repertoire of activating receptors and the cytokine profile of NK cells, rendering tumor cells more susceptible to NK cell-mediated lysis through the activation of NKG2D and DNAM-1 receptors. In addition, hyperploidization stimulated the production of IL-2 by CD4 T cells, which induced NK cell proliferation and activity. The stimulation of MICA, a key NKG2D ligand, in hyperploid cells was mainly mediated by ATM protein kinase. Likewise, pharmacological inhibition of key regulators of endoplasmic reticulum stress in certain cell models supports a role for this pathway in NKG2D ligand upregulation. Overall, our findings indicate that, besides the cytotoxic effect on tumor cells, the therapeutic activity of anti-mitotic drugs may be mediated by the induction of a coordinated antitumor immune response involving NK and T cells. PMID:27057443

  3. Development of drug loaded nanoparticles for tumor targeting. Part 1: synthesis, characterization, and biological evaluation in 2D cell cultures

    NASA Astrophysics Data System (ADS)

    El-Dakdouki, Mohammad H.; Puré, Ellen; Huang, Xuefei

    2013-04-01

    significantly enhanced through receptor mediated endocytosis. In addition, if the receptor is recycled to the cell surface, the NP cargo can be transported out of the cells, which is then taken up by neighboring cells thus enhancing solid tumor penetration. To validate our hypothesis, in the first of two articles, we report the synthesis of doxorubicin (DOX)-loaded, hyaluronan (HA) coated silica nanoparticles (SNPs) containing a highly fluorescent core to target CD44, a receptor expressed on the cancer cell surface. HA was conjugated onto amine-functionalized SNPs prepared through an oil-water microemulsion method. The immobilization of the cytotoxic drug DOX was achieved through an acid sensitive hydrazone linkage. The NPs were fully characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential measurements, thermogravimetric analysis (TGA), UV-vis absorbance, and nuclear magnetic resonance (NMR). Initial biological evaluation experiments demonstrated that compared to ligand-free SNPs, the uptake of HA-SNPs by the CD44-expressing SKOV-3 ovarian cancer cells was significantly enhanced when evaluated in the 2D monolayer cell culture. Mechanistic studies suggested that cellular uptake of HA-SNPs was mainly through CD44 mediated endocytosis. HA-SNPs with immobilized DOX were endocytosed efficiently by the SKOV-3 cells as well. The enhanced tumor penetration and drug delivery properties of HA-SNPs will be evaluated in 3D tumor models in the subsequent paper. Electronic supplementary information (ESI) available: Detailed experimental procedures for the synthesis and characterization of nanoparticles, confocal studies, flow cytometry, and supporting figures and tables. See DOI: 10.1039/c3nr33777d

  4. Myoblast alignment on 2D wavy patterns: dependence on feature characteristics and cell-cell interaction.

    PubMed

    Grigola, Michael S; Dyck, Casey L; Babacan, Derin S; Joaquin, Danielle N; Hsia, K Jimmy

    2014-08-01

    In this study, we investigate the effects of micron-scale surface patterns on the alignment of individual cells and groups of cells. Using a simple replication molding process we produce a number of micron-scale periodic wavy patterns with different pitch and depth. We observe C2C12 cells as they grow to confluence on these patterns and find that, for some geometries, cell-cell interaction leads to global alignment in a confluent culture when individual cells would not align on the same pattern. Three types of alignment behavior are thus defined: no alignment, immediate alignment, and alignment upon confluence. To further characterize this response, we introduce a non-dimensional parameter that describes the aligning power of a periodic pattern based on its geometry. The three types of alignment behavior can be distinguished by the value of the alignment parameter, and we identify values at which the transitions in alignment behavior occur. Applying this parameter to data from the current and several earlier studies reveals that the parameter successfully describes substrate aligning power over a wide range of length scales for both wavy and grooved features.

  5. TITAN2D simulations of pyroclastic flows at Cerro Machín Volcano, Colombia: Hazard implications

    NASA Astrophysics Data System (ADS)

    Murcia, H. F.; Sheridan, M. F.; Macías, J. L.; Cortés, G. P.

    2010-03-01

    Cerro Machín is a dacitic tuff ring located in the central part of the Colombian Andes. It lies at the southern end of the Cerro Bravo-Cerro Machín volcanic belt. This volcano has experienced at least six major explosive eruptions during the last 5000 years. These eruptions have generated pyroclastic flows associated with Plinian activity that have traveled up to 8 km from the crater, and pyroclastic flows associated with Vulcanian activity with shorter runouts of 5 km from the source. Today, some 21,000 people live within a 8 km radius of Cerro Machín. The volcano is active with fumaroles and has shown increasing seismic activity since 2004, and therefore represents a potentially increasing threat to the local population. To evaluate the possible effects of future eruptions that may generate pyroclastic density currents controlled by granular flow dynamics we performed flow simulations with the TITAN2D code. These simulations were run in all directions around the volcano, using the input parameters of the largest eruption reported. The results show that an eruption of 0.3 km 3 of pyroclastic flows from a collapsing Plinian column would travel up to 9 km from the vent, emplacing a deposit thicker than 60 m within the Toche River valley. Deposits >45 m thick can be expected in the valleys of San Juan, Santa Marta, and Azufral creeks, while 30 m thick deposits could accumulate within the drainages of the Tochecito, Bermellón, and Coello Rivers. A minimum area of 56 km 2 could be affected directly by this kind of eruption. In comparison, Vulcanian column-collapse pyroclastic flows of 0.1 km 3 would travel up to 6 km from the vent depositing >45 m thick debris inside the Toche River valley and more than 30 m inside the valleys of San Juan, Santa Marta, and Azufral creeks. The minimum area that could be affected directly by this kind of eruption is 33 km 2. The distribution and thickness of the deposits obtained by these simulations are consistent with the hazard

  6. 1α,25(OH)2D3 differentially regulates miRNA expression in human bladder cancer cells.

    PubMed

    Ma, Yingyu; Hu, Qiang; Luo, Wei; Pratt, Rachel N; Glenn, Sean T; Liu, Song; Trump, Donald L; Johnson, Candace S

    2015-04-01

    Bladder cancer is the fourth most commonly diagnosed cancer in men and eighth leading cause of cancer-related death in the US. Epidemiological and experimental studies strongly suggest a role for 1α,25(OH)2D3 in cancer prevention and treatment. The antitumor activities of 1α,25(OH)2D3 are mediated by the induction of cell cycle arrest, apoptosis, differentiation and the inhibition of angiogenesis and metastasis. miRNAs play important regulatory roles in cancer development and progression. However, the role of 1α,25(OH)2D3 in the regulation of miRNA expression and the potential impact in bladder cancer has not been investigated. Therefore, we studied 1α,25(OH)2D3-regulated miRNA expression profiles in human bladder cancer cell line 253J and the highly tumorigenic and metastatic derivative line 253J-BV by miRNA qPCR panels. 253J and 253J-BV cells express endogenous vitamin D receptor (VDR), which can be further induced by 1α,25(OH)2D3. VDR target gene 24-hydroxylase was induced by 1α,25(OH)2D3 in both cell lines, indicating functional 1α,25(OH)2D3 signaling. The miRNA qPCR panel assay results showed that 253J and 253J-BV cells have distinct miRNA expression profiles. Further, 1α,25(OH)2D3 differentially regulated miRNA expression profiles in 253J and 253J-BV cells in a dynamic manner. Pathway analysis of the miRNA target genes revealed distinct patterns of contribution to the molecular functions and biological processes in the two cell lines. In conclusion, 1α,25(OH)2D3 differentially regulates the expression of miRNAs, which may contribute to distinct biological functions, in human bladder 253J and 253J-BV cells. This article is part of a Special Issue entitled '17th Vitamin D Workshop'. PMID:25263658

  7. 1α,25(OH)2D3 differentially regulates miRNA expression in human bladder cancer cells.

    PubMed

    Ma, Yingyu; Hu, Qiang; Luo, Wei; Pratt, Rachel N; Glenn, Sean T; Liu, Song; Trump, Donald L; Johnson, Candace S

    2015-04-01

    Bladder cancer is the fourth most commonly diagnosed cancer in men and eighth leading cause of cancer-related death in the US. Epidemiological and experimental studies strongly suggest a role for 1α,25(OH)2D3 in cancer prevention and treatment. The antitumor activities of 1α,25(OH)2D3 are mediated by the induction of cell cycle arrest, apoptosis, differentiation and the inhibition of angiogenesis and metastasis. miRNAs play important regulatory roles in cancer development and progression. However, the role of 1α,25(OH)2D3 in the regulation of miRNA expression and the potential impact in bladder cancer has not been investigated. Therefore, we studied 1α,25(OH)2D3-regulated miRNA expression profiles in human bladder cancer cell line 253J and the highly tumorigenic and metastatic derivative line 253J-BV by miRNA qPCR panels. 253J and 253J-BV cells express endogenous vitamin D receptor (VDR), which can be further induced by 1α,25(OH)2D3. VDR target gene 24-hydroxylase was induced by 1α,25(OH)2D3 in both cell lines, indicating functional 1α,25(OH)2D3 signaling. The miRNA qPCR panel assay results showed that 253J and 253J-BV cells have distinct miRNA expression profiles. Further, 1α,25(OH)2D3 differentially regulated miRNA expression profiles in 253J and 253J-BV cells in a dynamic manner. Pathway analysis of the miRNA target genes revealed distinct patterns of contribution to the molecular functions and biological processes in the two cell lines. In conclusion, 1α,25(OH)2D3 differentially regulates the expression of miRNAs, which may contribute to distinct biological functions, in human bladder 253J and 253J-BV cells. This article is part of a Special Issue entitled '17th Vitamin D Workshop'.

  8. Miniaturized Electrochemical Flow Cells

    PubMed Central

    Sahlin, Eskil; Halle, Alexandra ter; Schaefer, Kathleen; Horn, Jeffery; Then, Matthew; Weber, Stephen G.

    2006-01-01

    Several novel types of miniaturized electrochemical flow cells are described. The flow cells are fabricated in fluorinated ethylene propylene using a novel technique where channels with inner diameters down to 13 μm are integrated with electrodes. The channel is formed by shrinking and simultaneous melting of a heat shrink/melt tubing around a channel template (a tungsten wire) and electrodes followed by removal of the channel template. The technique allows incorporation of different electrode materials of different sizes. The electrode configuration consists of one or two working electrodes inside the channel and a counter electrode located in the channel outlet reservoir. Electrode configurations with different channel and working electrode sizes, different electrode materials including carbon fibers, glassy carbon rods, poly(tetrafluoroethylene)/carbon composite material, and platinum wires, and different arrangements have been assembled. Hydrodynamic voltammograms in dual-electrode (generator–collector) experiments indicate good potential control for cells with 25-μm channels, while there is some iR drop in cells with 13-μm channels. Cells prepared with a cylindrical working electrode tangent and perpendicular to a flow channel show a flow rate dependence consistent with thin-layer cell behavior. Electrode areas can be made in the range of 10−10–10−8 m2. PMID:12622401

  9. Planar arrangement of eukaryotic cells in merged hydrogels combines the advantages of 3-D and 2-D cultures.

    PubMed

    Gordeev, Alexander A; Chetverina, Helena V; Chetverin, Alexander B

    2012-05-01

    We report an unordered 2-D array of eukaryotic cells completely embedded in a 3-D matrix. Every cell is located at the same distance from the gel surface, which ensures uniformity of growth conditions and ease of observation characteristic of a 2-D culture. Each cell is firmly immobilized, and each has a unique address in the array. The cells can be rapidly screened, individually monitored during extended time periods, and cultured with the formation of spheroid microcolonies characteristic of a 3-D culture. Individual microcolonies can be extracted from the gel and further propagated, thus enabling isolation of pure cell clones from rather dense cell populations and rapid drug-free generation of stable cell lines.

  10. Cytomegalovirus-Associated CD4(+) CD28(null) Cells in NKG2D-Dependent Glomerular Endothelial Injury and Kidney Allograft Dysfunction.

    PubMed

    Shabir, S; Smith, H; Kaul, B; Pachnio, A; Jham, S; Kuravi, S; Ball, S; Chand, S; Moss, P; Harper, L; Borrows, R

    2016-04-01

    Emerging data suggest that expansion of a circulating population of atypical, cytotoxic CD4(+) T cells lacking costimulatory CD28 (CD4(+) CD28(null) cells) is associated with latent cytomegalovirus (CMV) infection. The purpose of the current study was to increase the understanding of the relevance of these cells in 100 unselected kidney transplant recipients followed prospectively for a median of 54 months. Multicolor flow cytometry of peripheral blood mononuclear cells before transplantation and serially posttransplantation was undertaken. CD4(+) CD28(null) cells were found predominantly in CMV-seropositive patients and expanded in the posttransplantation period. These cells were predominantly effector-memory phenotype and expressed markers of endothelial homing (CX3CR1) and cytotoxicity (NKG2D and perforin). Isolated CD4(+) CD27(-) CD28(null) cells proliferated in response to peripheral blood mononuclear cells previously exposed to CMV-derived (but not HLA-derived) antigens and following such priming incubation with glomerular endothelium resulted in signs of endothelial damage and apoptosis (release of fractalkine and von Willebrand factor; increased caspase 3 expression). This effect was mitigated by NKG2D-blocking antibody. Increased CD4(+) CD28(null) cell frequencies were associated with delayed graft function and lower estimated glomerular filtration rate at end follow-up. This study suggests an important role for this atypical cytotoxic CD4(+) CD28(null) cell subset in kidney transplantation and points to strategies that may minimize the impact on clinical outcomes.

  11. 1-integrin and MT1-MMP promote tumor cell migration in 2D but not in 3D fibronectin microenvironments

    NASA Astrophysics Data System (ADS)

    Corall, Silke; Haraszti, Tamas; Bartoschik, Tanja; Spatz, Joachim Pius; Ludwig, Thomas; Cavalcanti-Adam, Elisabetta Ada

    2014-03-01

    Cell migration is a crucial event for physiological processes, such as embryonic development and wound healing, as well as for pathological processes, such as cancer dissemination and metastasis formation. Cancer cell migration is a result of the concerted action of matrix metalloproteinases (MMPs), expressed by cancer cells to degrade the surrounding matrix, and integrins, the transmembrane receptors responsible for cell binding to matrix proteins. While it is known that cell-microenvironment interactions are essential for migration, the role of the physical state of such interactions remains still unclear. In this study we investigated human fibrosarcoma cell migration in two-dimensional (2D) and three-dimensional (3D) fibronectin (FN) microenvironments. By using antibody blocking approach and cell-binding site mutation, we determined that -integrin is the main mediator of fibrosarcoma cell migration in 2D FN, whereas in 3D fibrillar FN, the binding of - and -integrins is not necessary for cell movement in the fibrillar network. Furthermore, while the general inhibition of MMPs with GM6001 has no effect on cell migration in both 2D and 3D FN matrices, we observed opposing effect after targeted silencing of a membrane-bound MMP, namely MT1-MMP. In 2D fibronectin, silencing of MT1-MMP results in decreased migration speed and loss of directionality, whereas in 3D FN matrices, cell migration speed is increased and integrin-mediated signaling for actin dynamics is promoted. Our results suggest that the fibrillar nature of the matrix governs the migratory behavior of fibrosarcoma cells. Therefore, to hinder migration and dissemination of diseased cells, matrix molecules should be directly targeted, rather than specific subtypes of receptors at the cell membrane.

  12. Differences in growth properties of endometrial cancer in three dimensional (3D) culture and 2D cell monolayer

    SciTech Connect

    Chitcholtan, Kenny; Asselin, Eric; Parent, Sophie; Sykes, Peter H.; Evans, John J.

    2013-01-01

    Three-dimensional (3D) in vitro models have an invaluable role in understanding the behaviour of tumour cells in a well defined microenvironment. This is because some aspects of tumour characteristics cannot be fully recapitulated in a cell monolayer (2D). In the present study, we compared growth patterns, expression of signalling molecules, and metabolism-associated proteins of endometrial cancer cell lines in 3D and 2D cell cultures. Cancer cells formed spherical structures in 3D reconstituted basement membrane (3D rBM), and the morphological appearance was cell line dependent. Cell differentiation was observed after 8 days in the 3D rBM. There was reduced proliferation, detected by less expression of PCNA in 3D rBM than in 2D cell monolayers. The addition of exogenous epidermal growth factor (EGF) to cancer cells induced phosphorylation of EGFR and Akt in both cell culture conditions. The uptake of glucose was selectively altered in the 3D rBM, but there was a lack of association with Glut-1 expression. The secretion of vascular endothelial growth factor (VEGF) and prostaglandin E{sub 2} (PGE{sub 2}) was selectively altered in 3D rBM, and it was cell line dependent. Our data demonstrated that 3D rBM as an in vitro model can influence proliferation and metabolism of endometrial cancer cell behaviour compared to 2D cell monolayer. Changes are specific to individual cell types. The use of 3D rBM is, therefore, important in the in vitro study of targeted anticancer therapies.

  13. Observation and simulation of heterogeneous 2D water and solute flow processes in ditch beds for subsequent catchment modelling

    NASA Astrophysics Data System (ADS)

    Dages, Cecile; Samouelian, Anatja; Lanoix, Marthe; Dollinger, Jeanne; Chakkour, Sara; Chovelon, Gabrielle; Trabelsi, Khouloud; Voltz, Marc

    2015-04-01

    Ditches are involved in the transfer of pesticide to surface and groundwaters (e.g. Louchart et al., 2001). Soil horizons underlying ditch beds may present specific soil characteristics compared to neighbouring field soils due to erosion/deposition processes, to the specific biological activities (rooting dynamic and animal habitat) in the ditches (e.g. Vaughan et al., 2008) and to management practices (burning, dredging, mowing,...). Moreover, in contrast to percolation processes in field soils that can be assumed to be mainly 1D vertical, those occurring in the ditch beds are by essence 2D or even 3D. Nevertheless, due to a lake of knowledge, these specific aspects of transfer within ditch beds are generally omitted for hydrological simulation at the catchment scale (Mottes et al., 2014). Accordingly, the aims of this study were i) to characterize subsurface solute transfer through ditch beds and ii) to determine equivalent hydraulic parameters of the ditch beds for use in catchment scale hydrological simulations. A complementary aim was to evaluate the error in predictions performed when percolation in ditches is assumed to be similar to that in the neighbouring field soil. First, bromide transfer experiments were performed on undisturbed soil column (15 cm long with a 15 cm inner-diameter), horizontally and vertically sampled within each soil horizon underlying a ditch bed and within the neighboring field. Columns were sampled at the Roujan catchment (Hérault, France), which belongs to the long term Mediterranean hydrological observatory OMERE (Voltz and Albergel, 2002). Second, for each column, a set of parameters was determined by inverse optimization with mobile-immobile or dual permeability models, with CXTFIT (Toride et al., 1999) or with HYDRUS (Simunek et al., 1998). Third, infiltration and percolation in the ditch was simulated by a 2D flow domain approach considering the 2D variation in hydraulic properties of the cross section of a ditch bed. Last

  14. Analytical description of 2D magnetic Freedericksz transition in a rectangular cell of a nematic liquid crystal.

    PubMed

    Burylov, S V; Zakhlevnykh, A N

    2016-06-01

    We study the Freedericksz transition induced by a magnetic field in a rectangular cell filled with a nematic liquid crystal. In the initial state the director of the nematic liquid crystal is uniformly aligned in the cross section plane of the cell with rigid anchoring of the director at cell walls: planar on the top and bottom walls, and homeotropic on the left and right ones. The magnetic field is directed perpendicular to the cell cross section plane. We consider two-dimensional (2D) orientational deformations of the nematic liquid crystal in the rectangular cell and determine the critical value of the Freedericksz transition field above which these orientational deformations occur. The 2D expression for the director alignment profile above the threshold of Freedericksz transition is analytically found and the profile shapes as functions of cell sizes, values of the Frank elastic constants of the nematic liquid crystal and the magnetic field are studied. PMID:27349554

  15. 2D design rule and layout analysis using novel large-area first-principles-based simulation flow incorporating lithographic and stress effects

    NASA Astrophysics Data System (ADS)

    Prins, Steven L.; Blatchford, James; Olubuyide, Oluwamuyiwa; Riley, Deborah; Chang, Simon; Hong, Qi-Zhong; Kim, T. S.; Borges, Ricardo; Lin, Li

    2009-03-01

    As design rules and corresponding logic standard cell layouts continue to shrink node-on-node in accordance with Moore's law, complex 2D interactions, both intra-cell and between cells, become much more prominent. For example, in lithography, lack of scaling of λ/NA implies aggressive use of resolution enhancement techniques to meet logic scaling requirements-resulting in adverse effects such as 'forbidden pitches'-and also implies an increasing range of optical influence relative to cell size. These adverse effects are therefore expected to extend well beyond the cell boundary, leading to lithographic marginalities that occur only when a given cell is placed "in context" with other neighboring cells in a variable design environment [1]. This context dependence is greatly exacerbated by increased use of strain engineering techniques such as SiGe and dual-stress liners (DSL) to enhance transistor performance, both of which also have interaction lengths on the order of microns. The use of these techniques also breaks the formerly straightforward connection between lithographic 'shapes' and end-of-line electrical performance, thus making the formulation of design rules that are robust to process variations and complex 2D interactions more difficult. To address these issues, we have developed a first-principles-based simulation flow to study contextdependent electrical effects in layout, arising not only from lithography, but also from stress and interconnect parasitic effects. This flow is novel in that it can be applied to relatively large layout clips- required for context-dependent analysis-without relying on semi-empirical or 'black-box' models for the fundamental electrical effects. The first-principles-based approach is ideal for understanding contextdependent effects early in the design phase, so that they can be mitigated through restrictive design rules. The lithographic simulations have been discussed elsewhere [1] and will not be presented in detail. The

  16. Numerical model of water flow and solute accumulation in vertisols using HYDRUS 2D/3D code

    NASA Astrophysics Data System (ADS)

    Weiss, Tomáš; Dahan, Ofer; Turkeltub, Tuvia

    2015-04-01

    Keywords: dessication-crack-induced-salinization, preferential flow, conceptual model, numerical model, vadose zone, vertisols, soil water retention function, HYDRUS 2D/3D Vertisols cover a hydrologically very significant area of semi-arid regions often through which water infiltrates to groundwater aquifers. Understanding of water flow and solute accumulation is thus very relevant to agricultural activity and water resources management. Previous works suggest a conceptual model of dessication-crack-induced-salinization where salinization of sediment in the deep section of the vadose zone (up to 4 m) is induced by subsurface evaporation due to convective air flow in the dessication cracks. It suggests that the salinization is induced by the hydraulic gradient between the dry sediment in the vicinity of cracks (low potential) and the relatively wet sediment further from the main cracks (high potential). This paper presents a modified previously suggested conceptual model and a numerical model. The model uses a simple uniform flow approach but unconventionally prescribes the boundary conditions and the hydraulic parameters of soil. The numerical model is bound to one location close to a dairy farm waste lagoon, but the application of the suggested conceptual model could be possibly extended to all semi-arid regions with vertisols. Simulations were conducted using several modeling approaches with an ultimate goal of fitting the simulation results to the controlling variables measured in the field: temporal variation in water content across thick layer of unsaturated clay sediment (>10 m), sediment salinity and salinity the water draining down the vadose zone to the water table. The development of the model was engineered in several steps; all computed as forward solutions by try-and-error approach. The model suggests very deep instant infiltration of fresh water up to 12 m, which is also supported by the field data. The paper suggests prescribing a special atmospheric

  17. Ganoderma lucidum stimulates NK cell cytotoxicity by inducing NKG2D/NCR activation and secretion of perforin and granulysin.

    PubMed

    Chang, Chih-Jung; Chen, Yi-Yuan M; Lu, Chia-Chen; Lin, Chuan-Sheng; Martel, Jan; Tsai, Sheng-Hui; Ko, Yun-Fei; Huang, Tsung-Teng; Ojcius, David M; Young, John D; Lai, Hsin-Chih

    2014-04-01

    Ganoderma lucidum (G. lucidum) is a medicinal mushroom long used in Asia as a folk remedy to promote health and longevity. Recent studies indicate that G. lucidum activates NK cells, but the molecular mechanism underlying this effect has not been studied so far. To address this question, we prepared a water extract of G. lucidum and examined its effect on NK cells. We observed that G. lucidum treatment increases NK cell cytotoxicity by stimulating secretion of perforin and granulysin. The mechanism of activation involves an increased expression of NKG2D and natural cytotoxicity receptors (NCRs), as well as increased phosphorylation of intracellular MAPKs. Our results indicate that G. lucidum induces NK cell cytotoxicity against various cancer cell lines by activating NKG2D/NCR receptors and MAPK signaling pathways, which together culminate in exocytosis of perforin and granulysin. These observations provide a cellular and molecular mechanism to account for the reported anticancer effects of G. lucidum extracts in humans.

  18. Ellipsoidal cell flow system

    DOEpatents

    Salzman, Gary C.; Mullaney, Paul F.

    1976-01-01

    The disclosure relates to a system incorporating an ellipsoidal flow chamber having light reflective walls for low level light detection in practicing cellular analysis. The system increases signal-to-noise ratio by a factor of ten over prior art systems. In operation, laser light passes through the primary focus of the ellipsoid. A controlled flow of cells simultaneously passes through this focus so that the laser light impinges on the cells and is modulated by the cells. The reflective walls of the ellipsoid reflect the cell-modulated light to the secondary focus of the ellipsoid. A tapered light guide at the secondary focus picks up a substantial portion of modulated reflective light and directs it onto a light detector to produce a signal. The signal is processed to obtain the intensity distribution of the modulated light and hence sought after characteristics of the cells. In addition, cells may be dyed so as to fluoresce in response to the laser light and their fluorescence may be processed as cell-modulated light above described. A light discriminating filter would be used to distinguish reflected modulated laser light from reflected fluorescent light.

  19. Study on the Multi-phase Flow and Fluid Saturation in 2D Fractured Media by Light Transmission Technique

    NASA Astrophysics Data System (ADS)

    Zhang, Y.; Ye, S.; Wu, J.

    2013-12-01

    Immiscible two-phase flows in fractured media are encountered in many engineering processes such as recovery of oil and gas, exploitation of geothermal energy, and groundwater contamination by immiscible chemicals. A two-dimensional rough wall parallel plate fracture model was set up and light transmission method (LTM) was applied to study two-phase flow system in fractured media. The fracture model stood with up and bottom flow and no flow on other two sides. A charge-coupled device (CCD) camera was used to monitor the migration of DNAPL and gas bubbles in the fracture model. To simulate two-phase system in fracture media, air was injected into the water saturated cell (C1) through the middle of the bottom and NAPL was injected into another water saturated cell (C2) through the middle of the top of the cell. The results show LTM was an effective way in monitoring the migration of DNAPL and gas bubbles in the fracture models. Gas moved upwards quickly to the top of C1 in the way of air bubbles generated at the injection position and formed a continuous distribution. The migration of TCE was controlled by its own weight and fracture aperture. TCE migrated to large aperture firstly when moving downwards, and intruded into smaller one with accumulation of TCE. Light Intensity-Saturation Models (LISMs) were developed to estimate the gas or NAPL saturation in two-phase system. The volume amount of infiltration of gas bubbles or NAPL could be estimated from light intensities by LISMs. There were strong correlations between the added and calculated amounts of gas or TCE. It is feasible to use the light transmission method to characterize the movement and spatial distribution of gas or NAPL in fractured media.

  20. Experimental investigation of compound-specific dilution of solute plumes in saturated porous media: 2-D vs. 3-D flow-through systems.

    PubMed

    Ye, Yu; Chiogna, Gabriele; Cirpka, Olaf; Grathwohl, Peter; Rolle, Massimo

    2015-01-01

    Dilution of solute plumes in groundwater strongly depends on transverse mixing. Thus, the correct parameterization of transverse dispersion is of critical importance for the quantitative description of solute transport. In this study we perform flow-through laboratory experiments to investigate the influence of transport dimensionality on transverse mixing. We present a high-resolution experimental setup to study solute dilution and transverse dispersion in three-dimensional porous media. We conduct multi-tracer experiments in the new 3-D setup and compare the results with the outcomes of analogous tracer experiments performed in a quasi 2-D system. We work under steady-state flow and transport conditions and consider a range of velocities relevant for groundwater flow (0.5-8 m/day). Transverse dispersion coefficients are determined from high-resolution concentration profiles at the outlet of the flow-through chambers (7×7 ports in the 3-D setup and 7 ports in the quasi 2-D system), considering conservative tracers with significantly different aqueous diffusion coefficients, namely fluorescein and dissolved oxygen. To quantify dilution in the 2-D and 3-D systems, we experimentally determine the flux-related dilution index using the flow rates and the concentrations measured at the inlet and outlet ports, and we propose semi-analytical expressions to predict its evolution with travel distance in uniform groundwater flow. The experimental results in the quasi 2-D and 3-D flow-through systems are consistent and show a compound-specific behavior of the transverse dispersion coefficient and its non-linear dependence on the seepage velocity in both setups. The degree of dilution and the compound-specific effects of transverse dispersion are considerably more pronounced in 3-D than in quasi 2-D transport systems.

  1. Documentation of computer program VS2D to solve the equations of fluid flow in variably saturated porous media

    USGS Publications Warehouse

    Lappala, E.G.; Healy, R.W.; Weeks, E.P.

    1987-01-01

    This report documents FORTRAN computer code for solving problems involving variably saturated single-phase flow in porous media. The flow equation is written with total hydraulic potential as the dependent variable, which allows straightforward treatment of both saturated and unsaturated conditions. The spatial derivatives in the flow equation are approximated by central differences, and time derivatives are approximated either by a fully implicit backward or by a centered-difference scheme. Nonlinear conductance and storage terms may be linearized using either an explicit method or an implicit Newton-Raphson method. Relative hydraulic conductivity is evaluated at cell boundaries by using either full upstream weighting, the arithmetic mean, or the geometric mean of values from adjacent cells. Nonlinear boundary conditions treated by the code include infiltration, evaporation, and seepage faces. Extraction by plant roots that is caused by atmospheric demand is included as a nonlinear sink term. These nonlinear boundary and sink terms are linearized implicitly. The code has been verified for several one-dimensional linear problems for which analytical solutions exist and against two nonlinear problems that have been simulated with other numerical models. A complete listing of data-entry requirements and data entry and results for three example problems are provided. (USGS)

  2. 2D dual permeability modeling of flow and transport in a two-scale structured lignitic mine soil

    NASA Astrophysics Data System (ADS)

    Dusek, J.; Gerke, H. H.; Vogel, T.; Maurer, T.; Buczko, U.

    2009-04-01

    Two-dimensional single- and dual-permeability simulations are used to analyze water and solute fluxes in heterogeneous lignitic mine soil at a forest-reclaimed mine spoil heap. The soil heterogeneity on this experimental site "Bärenbrücker Höhe" resulted from inclined dumping structures and sediment mixtures that consist of sand with lignitic dust and embedded lignitic fragments. Observations on undisturbed field suction-cell lysimeters including tracer experiments revealed funneling-type preferential flow with lateral water and bromide movement along inclined sediment structures. The spatial distribution of soil structures and fragment distributions was acquired by a digital camera and identified by a supervised classification of the digital profile image. First, a classical single-domain modeling approach was used, with spatially variable scaling factors inferred from image analyses. In the next step, a two-continuum scenario was constructed to examine additional effects of nonequilibrium on the flow regime. The scaling factors used for the preferential flow domain are here obtained from the gradient of the grayscale images. So far, the single domain scenarios failed to predict the bromide leaching patterns although water effluent could be described. Dual-permeability model allows the incorporation of structural effects and can be used as a tool to further testing other approaches that account for structure effects. The numerical study suggests that additional experiments are required to obtain better understanding of the highly complex transport processes on this experimental site.

  3. STAT3 contributes to NK cell recognition by modulating expression of NKG2D ligands in adriamycin-resistant K562/AO2 cells.

    PubMed

    Cai, Xiaohui; Lu, Xuzhang; Jia, Zhuxia; Zhang, Xiuwen; Han, Wenmin; Rong, Xiao; Ma, Lingdi; Zhou, Min; Chen, Baoan

    2015-11-01

    Leukemic cells can survive after chemotherapy by acquisition of multidrug resistance genes, but other phenotypes related to escape from immune recognition remain elusive. Adriamycin-resistant K562/AO2 cells are less susceptible to elimination by NK cells compared with wild type K562 cells due to lower expression of NKG2D ligands. Treatment of K562/AO2 cells with STAT3 inhibitor VII resulted in reduced expression of multidrug resistance gene P-glycoprotein, and up-regulation of NKG2D ligands on K562/AO2 cells. Meanwhile, K562/AO2 cells treated with STAT3 inhibitor proliferated less and were more susceptible to killing by NK cells than untreated K562/AO2 cells. The enhanced cytotoxicity of NK cells against K562/AO2 cells was partly blocked by treatment of NK cells with anti-NKG2D antibodies. These data suggest that STAT3 contributes to NK cell recognition by modulating NKG2D ligands in K562/AO2 cells, which may a mechanism by which cells survive and cause relapse of leukemia.

  4. NKG2D- and T-cell receptor-dependent lysis of malignant glioma cell lines by human γδ T cells: Modulation by temozolomide and A disintegrin and metalloproteases 10 and 17 inhibitors

    PubMed Central

    Chitadze, Guranda; Lettau, Marcus; Luecke, Stefanie; Wang, Ting; Janssen, Ottmar; Fürst, Daniel; Mytilineos, Joannis; Wesch, Daniela; Oberg, Hans-Heinrich; Held-Feindt, Janka; Kabelitz, Dieter

    2016-01-01

    ABSTRACT The interaction of the MHC class I-related chain molecules A and B (MICA and MICB) and UL-16 binding protein (ULBP) family members expressed on tumor cells with the corresponding NKG2D receptor triggers cytotoxic effector functions in NK cells and γδ T cells. However, as a mechanism of tumor immune escape, NKG2D ligands (NKG2DLs) can be released from the cell surface. In this study, we investigated the NKG2DL system in different human glioblastoma (GBM) cell lines, the most lethal brain tumor in adults. Flow cytometric analysis and ELISA revealed that despite the expression of various NKG2DLs only ULBP2 is released as a soluble protein via the proteolytic activity of “a disintegrin and metalloproteases” (ADAM) 10 and 17. Moreover, we report that temozolomide (TMZ), a chemotherapeutic agent in clinical use for the treatment of GBM, increases the cell surface expression of NKG2DLs and sensitizes GBM cells to γδ T cell-mediated lysis. Both NKG2D and the T-cell receptor (TCR) are involved. The cytotoxic activity of γδ T cells toward GBM cells is strongly enhanced in a TCR-dependent manner by stimulation with pyrophosphate antigens. These data clearly demonstrate the complexity of mechanisms regulating NKG2DL expression in GBM cells and further show that treatment with TMZ can increase the immunogenicity of GBM. Thus, TMZ might enhance the potential of the adoptive transfer of ex vivo expanded γδ T cells for the treatment of malignant glioblastoma. PMID:27141377

  5. Redirecting T cells to Ewing's sarcoma family of tumors by a chimeric NKG2D receptor expressed by lentiviral transduction or mRNA transfection.

    PubMed

    Lehner, Manfred; Götz, Gabriel; Proff, Julia; Schaft, Niels; Dörrie, Jan; Full, Florian; Ensser, Armin; Muller, Yves A; Cerwenka, Adelheid; Abken, Hinrich; Parolini, Ornella; Ambros, Peter F; Kovar, Heinrich; Holter, Wolfgang

    2012-01-01

    We explored the possibility to target Ewing's sarcoma family of tumors (ESFT) by redirecting T cells. To this aim, we considered NKG2D-ligands (NKG2D-Ls) as possible target antigens. Detailed analysis of the expression of MICA, MICB, ULBP-1, -2, and -3 in fourteen ESFT cell lines revealed consistent expression of at least one NKG2D-L. Thus, for redirecting T cells, we fused a CD3ζ/CD28-derived signaling domain to the ectodomain of NKG2D, however, opposite transmembrane orientation of this signaling domain and NKG2D required inverse orientation fusion of either of them. We hypothesized that the particularly located C-terminus of the NKG2D ectodomain should allow reengineering of the membrane anchoring from a native N-terminal to an artificial C-terminal linkage. Indeed, the resulting chimeric NKG2D receptor (chNKG2D) was functional and efficiently mediated ESFT cell death triggered by activated T cells. Notably, ESFT cells with even low NKG2D-L expression were killed by CD8(pos) and also CD4(pos) cells. Both, mRNA transfection and lentiviral transduction resulted in high level surface expression of chNKG2D. However, upon target-cell recognition receptor surface levels were maintained by tranfected RNA only during the first couple of hours after transfection. Later, target-cell contact resulted in strong and irreversible receptor down-modulation, whereas lentivirally mediated expression of chNKG2D remained constant under these conditions. Together, our study defines NKG2D-Ls as targets for a CAR-mediated T cell based immunotherapy of ESFT. A comparison of two different methods of gene transfer reveals strong differences in the susceptibility to ligand-induced receptor down-modulation with possible implications for the applicability of RNA transfection. PMID:22355347

  6. A 2-D FEM thermal model to simulate water flow in a porous media: Campi Flegrei caldera case study

    NASA Astrophysics Data System (ADS)

    Romano, V.; Tammaro, U.; Capuano, P.

    2012-05-01

    Volcanic and geothermal aspects both exist in many geologically young areas. In these areas the heat transfer process is of fundamental importance, so that the thermal and fluid-dynamic processes characterizing a viscous fluid in a porous medium are very important to understand the complex dynamics of the these areas. The Campi Flegrei caldera, located west of the city of Naples, within the central-southern sector of the large graben of Campanian plain, is a region where both volcanic and geothermal phenomena are present. The upper part of the geothermal system can be considered roughly as a succession of volcanic porous material (tuff) saturated by a mixture formed mainly by water and carbon dioxide. We have implemented a finite elements approach in transient conditions to simulate water flow in a 2-D porous medium to model the changes of temperature in the geothermal system due to magmatic fluid inflow, accounting for a transient phase, not considered in the analytical solutions and fluid compressibility. The thermal model is described by means of conductive/convective equations, in which we propose a thermal source represented by a parabolic shape function to better simulate an increase of temperature in the central part (magma chamber) of a box, simulating the Campi Flegrei caldera and using more recent evaluations, from literature, for the medium's parameters (specific heat capacity, density, thermal conductivity, permeability). A best-fit velocity for the permeant is evaluated by comparing the simulated temperatures with those measured in wells drilled by Agip (Italian Oil Agency) in the 1980s in the framework of geothermal exploration. A few tens of days are enough to reach the thermal steady state, showing the quick response of the system to heat injection. The increase in the pressure due to the heat transport is then used to compute ground deformation, in particular the vertical displacements characteristics of the Campi Flegrei caldera behaviour. The

  7. Collagen esterification enhances the function and survival of pancreatic β cells in 2D and 3D culture systems

    SciTech Connect

    Ko, Jae Hyung; Kim, Yang Hee; Jeong, Seong Hee; Lee, Song; Park, Si-Nae; Shim, In Kyong; Kim, Song Cheol

    2015-08-07

    Collagen, one of the most important components of the extracellular matrix (ECM), may play a role in the survival of pancreatic islet cells. In addition, chemical modifications that change the collagen charge profile to a net positive charge by esterification have been shown to increase the adhesion and proliferation of various cell types. The purpose of this study was to characterize and compare the effects of native collagen (NC) and esterified collagen (EC) on β cell function and survival. After isolation by the collagenase digestion technique, rat islets were cultured with NC and EC in 2 dimensional (2D) and 3 dimensional (3D) environments for a long-term duration in vitro. The cells were assessed for islet adhesion, morphology, viability, glucose-induced insulin secretion, and mRNA expression of glucose metabolism-related genes, and visualized by scanning electron microscopy (SEM). Islet cells attached tightly in the NC group, but islet cell viability was similar in both the NC and EC groups. Glucose-stimulated insulin secretion was higher in the EC group than in the NC group in both 2D and 3D culture. Furthermore, the mRNA expression levels of glucokinase in the EC group were higher than those in the NC group and were associated with glucose metabolism and insulin secretion. Finally, SEM observation confirmed that islets had more intact component cells on EC sponges than on NC sponges. These results indicate that modification of collagen may offer opportunities to improve function and viability of islet cells. - Highlights: • We changed the collagen charge profile to a net positive charge by esterification. • Islets cultured on esterified collagen improved survival in both 2D and 3D culture. • Islets cultured on esterified collagen enhanced glucose-stimulated insulin release. • High levels of glucokinase mRNA may be associated with increased insulin release.

  8. Scorpion venom activates natural killer cells in hepatocellular carcinoma via the NKG2D-MICA pathway.

    PubMed

    Chen, Han; Zhidan, Wang; Xia, Ren; Zhaoxia, Wang; Qing, Jia; Qiang, Guo; Haipeng, Yin; Hengxiao, Wang

    2016-06-01

    Previous studies have demonstrated that polypeptides extracted from scorpion venom (PESV) inhibited cell proliferation in several tumors, however, the effect on dysfunctional and exhausted natural killer cells which contribute to tumor escape from immune surveillance remain to be elucidated. In this study, we determined the effect of PESV on NK infiltration into H22 cells orthotopic transplantation tumors and on the expression of MHC class I chain-related proteins A (MICA) in HepG2 cells. We found that tumor growth in mice was significantly inhibited by PESV and the survival time of tumor-bearing mice treated with PESV was significantly prolonged. Moreover, levels of tumor-infiltrating NK cells, NKG2D protein, perforin and granzyme B mRNA were significantly increased in the group treated with PESV compared with the tumor-bearing control group. In addition, In addition, up-regulation of MICA by PESV enhances the susceptibility of HepG2 cells to NK lysis in vitro. These results indicate that the inhibitory effects of PESV on hepatic carcinoma are likely mediated by up-regulation of NK cell activity via the MICA-NKG2D pathway. PMID:27089390

  9. Scorpion venom activates natural killer cells in hepatocellular carcinoma via the NKG2D-MICA pathway.

    PubMed

    Chen, Han; Zhidan, Wang; Xia, Ren; Zhaoxia, Wang; Qing, Jia; Qiang, Guo; Haipeng, Yin; Hengxiao, Wang

    2016-06-01

    Previous studies have demonstrated that polypeptides extracted from scorpion venom (PESV) inhibited cell proliferation in several tumors, however, the effect on dysfunctional and exhausted natural killer cells which contribute to tumor escape from immune surveillance remain to be elucidated. In this study, we determined the effect of PESV on NK infiltration into H22 cells orthotopic transplantation tumors and on the expression of MHC class I chain-related proteins A (MICA) in HepG2 cells. We found that tumor growth in mice was significantly inhibited by PESV and the survival time of tumor-bearing mice treated with PESV was significantly prolonged. Moreover, levels of tumor-infiltrating NK cells, NKG2D protein, perforin and granzyme B mRNA were significantly increased in the group treated with PESV compared with the tumor-bearing control group. In addition, In addition, up-regulation of MICA by PESV enhances the susceptibility of HepG2 cells to NK lysis in vitro. These results indicate that the inhibitory effects of PESV on hepatic carcinoma are likely mediated by up-regulation of NK cell activity via the MICA-NKG2D pathway.

  10. A microfluidic device for 2D to 3D and 3D to 3D cell navigation

    NASA Astrophysics Data System (ADS)

    Shamloo, Amir; Amirifar, Leyla

    2016-01-01

    Microfluidic devices have received wide attention and shown great potential in the field of tissue engineering and regenerative medicine. Investigating cell response to various stimulations is much more accurate and comprehensive with the aid of microfluidic devices. In this study, we introduced a microfluidic device by which the matrix density as a mechanical property and the concentration profile of a biochemical factor as a chemical property could be altered. Our microfluidic device has a cell tank and a cell culture chamber to mimic both 2D to 3D and 3D to 3D migration of three types of cells. Fluid shear stress is negligible on the cells and a stable concentration gradient can be obtained by diffusion. The device was designed by a numerical simulation so that the uniformity of the concentration gradients throughout the cell culture chamber was obtained. Adult neural cells were cultured within this device and they showed different branching and axonal navigation phenotypes within varying nerve growth factor (NGF) concentration profiles. Neural stem cells were also cultured within varying collagen matrix densities while exposed to NGF concentrations and they experienced 3D to 3D collective migration. By generating vascular endothelial growth factor concentration gradients, adult human dermal microvascular endothelial cells also migrated in a 2D to 3D manner and formed a stable lumen within a specific collagen matrix density. It was observed that a minimum absolute concentration and concentration gradient were required to stimulate migration of all types of the cells. This device has the advantage of changing multiple parameters simultaneously and is expected to have wide applicability in cell studies.

  11. Recognition and killing of cancer stem-like cell population in hepatocellular carcinoma cells by cytokine-induced killer cells via NKG2d-ligands recognition

    PubMed Central

    Rong, Xiao-Xiang; Wei, Fang; Lin, Xiao-Lin; Qin, Yu-Juan; Chen, Lin; Wang, Hui-Yan; Shen, Hong-Fen; Jia, Li-Ting; Xie, Rao-Ying; Lin, Tao-Yan; Hao, Wei-Chao; Yang, Jie; Yang, Sheng; Cheng, Yu-Shuang; Huang, Wen-Hua; Li, Ai-min; Sun, Yan; Luo, Rong-Cheng; Xiao, Dong

    2016-01-01

    ABSTRACT There is an urgent need for more potent and safer approaches to eradicate cancer stem cells (CSCs) for curing cancer. In this study, we investigate cancer-killing activity (CKA) of cytokine-induced killer (CIK) cells against CSCs of hepatocellular carcinoma (HCC). To visualize CSCs in vitro by fluorescence imaging, and image and quantify CSCs in tumor xenograft-bearing mice by bioluminescence imaging, HCC cells were engineered with CSC detector vector encoding GFP and luciferase controlled by Nanog promoter. We found that CIK cells have a strong CKA in vitro against putative CSCs of HCC, as shown by tumorsphere formation and time-lapse imaging. Additionally, time-lapse recording firstly revealed that putative CSCs were attacked simultaneously by many CIK cells and finally eradicated by CIK cells, indicating the necessity of achieving sufficient effector-to-target ratios. We firstly illustrated that anti-NKG2D antibody blocking partially but significantly inhibited CKA of CIK cells against putative CSCs. More importantly, intravenous infusion of CIK cells remarkably delayed tumor growth in mice with a significant decrease in putative CSC number monitored by bioluminescence imaging. Taken together, these findings demonstrate CKA of CIK cells against putative CSCs of HCC, at least in part, by NKG2D-ligands recognition. PMID:27141341

  12. Application of micro-PIV technique to study multiphase flow of water and liquid CO2 in 2D porous media

    NASA Astrophysics Data System (ADS)

    Kazemifar, F.; Blois, G.; Kyritsis, D. C.; Christensen, K. T.

    2014-11-01

    We study the multiphase flow of water and liquid/supercritical CO2 in 2D porous micromodels, with the goal of developing a more complete understanding of pore-scale flow dynamics for the scenario of geological sequestration of carbon dioxide. Fluorescent microscopy and the micro-PIV technique are employed to simultaneously visualize both phases and obtain the velocity field in the aqueous phase. This technique provides a powerful tool for studying such flow systems and the results give valuable insight into flow processes at the pore scale. The fluid-fluid interface curvature from the images can be used to estimate the local capillary pressure. The velocity measurements illustrate active and passive flow pathways and circulation regions near the fluid-fluid interfaces induced by shear. Thin water films observed on the solid surfaces confirm the hydrophilic nature of the micromodels. The velocity of the said films is measured by particle tracking.

  13. New insights from velocity field measurements in multiphase flow of water and liquid CO2 in 2D porous micromodels for

    NASA Astrophysics Data System (ADS)

    Kazemifar, F.; Blois, G.; Kyritsis, D. C.; Christensen, K. T.

    2014-12-01

    We study the multiphase flow of water and liquid/supercritical CO2 in 2D porous micromodels, with the goal of developing a more complete understanding of pore-scale flow dynamics for the scenario of geological sequestration of carbon dioxide. Fluorescent microscopy and the microscopic particle image velocimetry (micro-PIV) technique are employed to simultaneously visualize both phases and obtain the velocity field in the aqueous phase. This technique provides a powerful tool for studying such flow systems and the results give valuable insight into flow processes at the pore scale. The fluid-fluid interface curvature from the images can be used to estimate the local capillary pressure. The velocity measurements illustrate active and passive flow pathways and circulation regions near the fluid-fluid interfaces induced by shear. Thin water films observed on the solid surfaces confirm the hydrophilic nature of the micromodels. The velocity of the said films is measured by particle tracking.

  14. An application of the FLO-2D Model to debris-flow simulation - a case study of Shinfa village in southern Taiwan

    NASA Astrophysics Data System (ADS)

    Chen, J.-C.; Chuang, M.-R.; Jeng, C.-J.; Wang, J.-S.

    2012-04-01

    Taiwan is an island located in the subtropical zone where typhoons often bring heavy rainfall. Heavy rainfall, stream having steep slope, and weak geological condition resulted in a high susceptibility to debris flow. Especially, Typhoon Morakot struck southern Taiwan on August 8, 2009 with high rainfall intensity and accumulated rainfall as high as 2860 mm for 72 hours. Severe landslides and debris flow hazards were induced. In this work, debris-flow events caused by Typhoon Morakot in Shinfa Village of Liouguei District, where resulted in severe impacts to local communities, in southern Taiwan were selected for case study. A two-dimensional model (FLO-2D software) was used to simulate a debris flow, and the accuracy of the simulation, including flow depth, velocity, sediment, and inundation area, was analyzed in the case study. This study consists of three phases. In the first phase, debris flow data, including information on topography, rainfall and rheological parameters were compiled to establish a database of factors that influence debris flow. For the second phase, a numerical simulation was performed using FLO-2D with the results presented as area of debris-flow inundation, maximum deposit depth, and deposit volume. The simulation results were then compared with the aerial photos and the micro geomorphological study. Finally, suitable conditions for using this model and reasonable parameters needed for simulation are presented. In this study, parameters and processes needed for a numerical simulation method for debris flow routing and depositions are formulated to provide a reference for hazard zone mapping or debris-flow hazard mitigation.

  15. Collagen esterification enhances the function and survival of pancreatic β cells in 2D and 3D culture systems.

    PubMed

    Ko, Jae Hyung; Kim, Yang Hee; Jeong, Seong Hee; Lee, Song; Park, Si-Nae; Shim, In Kyong; Kim, Song Cheol

    2015-08-01

    Collagen, one of the most important components of the extracellular matrix (ECM), may play a role in the survival of pancreatic islet cells. In addition, chemical modifications that change the collagen charge profile to a net positive charge by esterification have been shown to increase the adhesion and proliferation of various cell types. The purpose of this study was to characterize and compare the effects of native collagen (NC) and esterified collagen (EC) on β cell function and survival. After isolation by the collagenase digestion technique, rat islets were cultured with NC and EC in 2 dimensional (2D) and 3 dimensional (3D) environments for a long-term duration in vitro. The cells were assessed for islet adhesion, morphology, viability, glucose-induced insulin secretion, and mRNA expression of glucose metabolism-related genes, and visualized by scanning electron microscopy (SEM). Islet cells attached tightly in the NC group, but islet cell viability was similar in both the NC and EC groups. Glucose-stimulated insulin secretion was higher in the EC group than in the NC group in both 2D and 3D culture. Furthermore, the mRNA expression levels of glucokinase in the EC group were higher than those in the NC group and were associated with glucose metabolism and insulin secretion. Finally, SEM observation confirmed that islets had more intact component cells on EC sponges than on NC sponges. These results indicate that modification of collagen may offer opportunities to improve function and viability of islet cells.

  16. Surface Acoustic Waves (SAW)-Based Biosensing for Quantification of Cell Growth in 2D and 3D Cultures

    PubMed Central

    Wang, Tao; Green, Ryan; Nair, Rajesh Ramakrishnan; Howell, Mark; Mohapatra, Subhra; Guldiken, Rasim; Mohapatra, Shyam Sundar

    2015-01-01

    Detection and quantification of cell viability and growth in two-dimensional (2D) and three-dimensional (3D) cell cultures commonly involve harvesting of cells and therefore requires a parallel set-up of several replicates for time-lapse or dose–response studies. Thus, developing a non-invasive and touch-free detection of cell growth in longitudinal studies of 3D tumor spheroid cultures or of stem cell regeneration remains a major unmet need. Since surface acoustic waves (SAWs) permit mass loading-based biosensing and have been touted due to their many advantages including low cost, small size and ease of assembly, we examined the potential of SAW-biosensing to detect and quantify cell growth. Herein, we demonstrate that a shear horizontal-surface acoustic waves (SH-SAW) device comprising two pairs of resonators consisting of interdigital transducers and reflecting fingers can be used to quantify mass loading by the cells in suspension as well as within a 3D cell culture platform. A 3D COMSOL model was built to simulate the mass loading response of increasing concentrations of cells in suspension in the polydimethylsiloxane (PDMS) well in order to predict the characteristics and optimize the design of the SH-SAW biosensor. The simulated relative frequency shift from the two oscillatory circuit systems (one of which functions as control) were found to be concordant to experimental data generated with RAW264.7 macrophage and A549 cancer cells. In addition, results showed that SAW measurements per se did not affect viability of cells. Further, SH-SAW biosensing was applied to A549 cells cultured on a 3D electrospun nanofiber scaffold that generate tumor spheroids (tumoroids) and the results showed the device's ability to detect changes in tumor spheroid growth over the course of eight days. Taken together, these results demonstrate the use of SH-SAW device for detection and quantification of cell growth changes over time in 2D suspension cultures and in 3D cell

  17. Surface Acoustic Waves (SAW)-Based Biosensing for Quantification of Cell Growth in 2D and 3D Cultures.

    PubMed

    Wang, Tao; Green, Ryan; Nair, Rajesh Ramakrishnan; Howell, Mark; Mohapatra, Subhra; Guldiken, Rasim; Mohapatra, Shyam Sundar

    2015-01-01

    Detection and quantification of cell viability and growth in two-dimensional (2D) and three-dimensional (3D) cell cultures commonly involve harvesting of cells and therefore requires a parallel set-up of several replicates for time-lapse or dose-response studies. Thus, developing a non-invasive and touch-free detection of cell growth in longitudinal studies of 3D tumor spheroid cultures or of stem cell regeneration remains a major unmet need. Since surface acoustic waves (SAWs) permit mass loading-based biosensing and have been touted due to their many advantages including low cost, small size and ease of assembly, we examined the potential of SAW-biosensing to detect and quantify cell growth. Herein, we demonstrate that a shear horizontal-surface acoustic waves (SH-SAW) device comprising two pairs of resonators consisting of interdigital transducers and reflecting fingers can be used to quantify mass loading by the cells in suspension as well as within a 3D cell culture platform. A 3D COMSOL model was built to simulate the mass loading response of increasing concentrations of cells in suspension in the polydimethylsiloxane (PDMS) well in order to predict the characteristics and optimize the design of the SH-SAW biosensor. The simulated relative frequency shift from the two oscillatory circuit systems (one of which functions as control) were found to be concordant to experimental data generated with RAW264.7 macrophage and A549 cancer cells. In addition, results showed that SAW measurements per se did not affect viability of cells. Further, SH-SAW biosensing was applied to A549 cells cultured on a 3D electrospun nanofiber scaffold that generate tumor spheroids (tumoroids) and the results showed the device's ability to detect changes in tumor spheroid growth over the course of eight days. Taken together, these results demonstrate the use of SH-SAW device for detection and quantification of cell growth changes over time in 2D suspension cultures and in 3D cell

  18. Surface Acoustic Waves (SAW)-Based Biosensing for Quantification of Cell Growth in 2D and 3D Cultures.

    PubMed

    Wang, Tao; Green, Ryan; Nair, Rajesh Ramakrishnan; Howell, Mark; Mohapatra, Subhra; Guldiken, Rasim; Mohapatra, Shyam Sundar

    2015-12-19

    Detection and quantification of cell viability and growth in two-dimensional (2D) and three-dimensional (3D) cell cultures commonly involve harvesting of cells and therefore requires a parallel set-up of several replicates for time-lapse or dose-response studies. Thus, developing a non-invasive and touch-free detection of cell growth in longitudinal studies of 3D tumor spheroid cultures or of stem cell regeneration remains a major unmet need. Since surface acoustic waves (SAWs) permit mass loading-based biosensing and have been touted due to their many advantages including low cost, small size and ease of assembly, we examined the potential of SAW-biosensing to detect and quantify cell growth. Herein, we demonstrate that a shear horizontal-surface acoustic waves (SH-SAW) device comprising two pairs of resonators consisting of interdigital transducers and reflecting fingers can be used to quantify mass loading by the cells in suspension as well as within a 3D cell culture platform. A 3D COMSOL model was built to simulate the mass loading response of increasing concentrations of cells in suspension in the polydimethylsiloxane (PDMS) well in order to predict the characteristics and optimize the design of the SH-SAW biosensor. The simulated relative frequency shift from the two oscillatory circuit systems (one of which functions as control) were found to be concordant to experimental data generated with RAW264.7 macrophage and A549 cancer cells. In addition, results showed that SAW measurements per se did not affect viability of cells. Further, SH-SAW biosensing was applied to A549 cells cultured on a 3D electrospun nanofiber scaffold that generate tumor spheroids (tumoroids) and the results showed the device's ability to detect changes in tumor spheroid growth over the course of eight days. Taken together, these results demonstrate the use of SH-SAW device for detection and quantification of cell growth changes over time in 2D suspension cultures and in 3D cell

  19. Easy fabrication of aligned PLLA nanofibers-based 2D scaffolds suitable for cell contact guidance studies.

    PubMed

    Mohanraj, John; Puzzi, Luca; Capria, Ennio; Corvaglia, Stefania; Casalis, Loredana; Mestroni, Luisa; Sbaizero, Orfeo; Fraleoni-Morgera, Alessandro

    2016-05-01

    An easy, low-cost and fast wet processing-based method named ASB-SANS (Auxiliary Solvent-Based Sublimation-Aided NanoStructuring) has been used to fabricate poly(l-lactic acid) (PLLA) highly ordered and hierarchically organized 2D fibrillar patterns, with fiber widths between 40 and 500 nm and lengths exceeding tens of microns. A clear contact guidance effect of these nanofibrillar scaffolds with respect to HeLa and NIH-3T3 cells growth has been observed, on top of an overall good viability. For NIH-3T3 pronounced elongation of the cells was observed, as well as a remarkable ability of the patterns to guide the extension of pseudopodia. Moreover, SEM imaging revealed filopodia stemming from both sides of the pseudopodia and aligned with the secondary PLLA nanofibrous structures created by the ASB-SANS procedure. These results validate ASB-SANS as a technique capable to provide biocompatible 2D nanofibrillar patterns suitable for studying phenomena of contact guidance (and, more in general, the behavior of cells onto nanofibrous scaffolds), at very low costs and in an extremely easy way, accessible to virtually any laboratory. PMID:26952427

  20. TPA decreases 1,25(OH)2D3 binding and calbindin D-28K in renal (MDBK) cells.

    PubMed

    Simboli-Campbell, M; Gagnon, A M; Franks, D J; Welsh, J

    1992-02-01

    The effect of the phorbol ester TPA (12-O-tetradecanoylphorbol 13-acetate) on vitamin D receptors (VDRs) was studied in MDBK cells, a normal bovine renal epithelial cell line. 24 h treatment of MDBK cells with TPA resulted in down-regulation of VDR number, with no change in the binding affinity for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) or approximate molecular weight determined by fast protein liquid chromatography (FPLC). TPA treatment also reduced the level of calbindin D-28K, a vitamin D-dependent renal protein. 4 alpha-Phorbol 12,13-didecanoate (4 alpha-PDD), an inactive phorbol ester, did not affect either 1,25(OH)2D3 binding or calbindin D-28K levels. TPA elicited a significant decrease in membrane-associated protein kinase C (PKC) activity which coincided with the reduction in VDR number and calbindin D-28K. These data support a link between TPA, PKC activity and vitamin D actions in kidney.

  1. Identification of the wave speed and the second viscosity of cavitation flows with 2D RANS computations - Part I

    NASA Astrophysics Data System (ADS)

    Decaix, J.; Alligné, S.; Nicolet, C.; Avellan, F.; Münch, C.

    2015-12-01

    1D hydro-electric models are useful to predict dynamic behaviour of hydro-power plants. Regarding vortex rope and cavitation surge in Francis turbines, the 1D models require some inputs that can be provided by numerical simulations. In this paper, a 2D cavitating Venturi is considered. URANS computations are performed to investigate the dynamic behaviour of the cavitation sheet depending on the frequency variation of the outlet pressure. The results are used to calibrate and to assess the reliability of the 1D models.

  2. Superdiffusive cell motility on 2D substrates modeled as a persistent Lévy walk

    NASA Astrophysics Data System (ADS)

    Passucci, Giuseppe; Brasch, Megan E.; Deakin, Nicholas O.; Turner, Christopher E.; Henderson, James H.; Manning, M. Lisa

    Cell motility is an essential part of many biological processes such as morphogenesis, wound healing and tumorigenesis. We quantified cell motility by tracking mouse fibroblast and human breast carcinoma nuclei to construct cell trajectories. The mean-squared displacement of these trajectories reveals that cell motion is super diffusive, where displacements scale faster than t 1 / 2 in all directions. Existing self-propelled particle (SPP) models that do not explicitly incorporate ensemble heterogeneity are unable to predict this super-diffusive behavior. Therefore we developed a run-and-tumble SPP model with Levy distributed run times that captures observed super-diffusive behavior in the mean-squared displacement as well as scaling collapse exponents of displacement probability distributions which match those of mouse fibroblast and human breast carcinoma cell trajectories. We additionally introduced small fluctuations in particle orientation during runs, which generates a crossover from super-diffusive to diffusive dynamics at a very long times. This timescale can be extracted in experiments from the velocity auto-correlation function, allowing us to explicitly test this model prediction.

  3. Cytochrome P450 2D6 enzyme neuroprotects against 1-methyl-4-phenylpyridinium toxicity in SH-SY5Y neuronal cells.

    PubMed

    Mann, Amandeep; Tyndale, Rachel F

    2010-04-01

    Cytochrome P450 (CYP) 2D6 is an enzyme that is expressed in liver and brain. It can inactivate neurotoxins such as 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, 1,2,3,4-tetrahydroisoquinoline and beta-carbolines. Genetically slow CYP2D6 metabolizers are at higher risk for developing Parkinson's disease, a risk that increases with exposure to pesticides. The goal of this study was to investigate the neuroprotective role of CYP2D6 in an in-vitro neurotoxicity model. SH-SY5Y human neuroblastoma cells express CYP2D6 as determined by western blotting, immunocytochemistry and enzymatic activity. CYP2D6 metabolized 3-[2-(N,N-diethyl-N-methylammonium)ethyl]-7-methoxy-4-methylcoumarin and the CYP2D6-specific inhibitor quinidine (1 microM) blocked 96 +/- 1% of this metabolism, indicating that CYP2D6 is functional in this cell line. Treatment of cells with CYP2D6 inhibitors (quinidine, propanolol, metoprolol or timolol) at varying concentrations significantly increased the neurotoxicity caused by 1-methyl-4-phenylpyridinium (MPP+) at 10 and 25 microM by between 9 +/- 1 and 22 +/- 5% (P < 0.01). We found that CYP3A is also expressed in SH-SY5Y cells and inhibiting CYP3A with ketoconazole significantly increased the cell death caused by 10 and 25 microM of MPP+ by between 8 +/- 1 and 30 +/- 3% (P < 0.001). Inhibiting both CYP2D6 and CYP3A showed an additive effect on MPP+ neurotoxicity. These data further support a possible role for CYP2D6 in neuroprotection from Parkinson's disease-causing neurotoxins, especially in the human brain where expression of CYP2D6 is high in some regions (e.g. substantia nigra). PMID:20345925

  4. Controlling integrin specificity and stem cell differentiation in 2-D and 3-D environments through regulation of fibronectin domain stability

    PubMed Central

    Martino, Mikaël M.; Mochizuki, Mayumi; Rothenfluh, Dominique A.; Rempel, Sandra A.; Hubbell, Jeffrey A.; Barker, Thomas H.

    2009-01-01

    The extracellular matrix (ECM) exerts powerful control over many cellular phenomena, including stem cell differentiation. As such, design and modulation of ECM analogs to ligate specific integrin is a promising approach to control cellular processes in vitro and in vivo for regenerative medicine strategies. Although fibronectin (FN), a crucial ECM protein in tissue development and repair, and its RGD peptide are widely used for cell adhesion, the promiscuity with which they engage integrins leads to difficulty in control of receptor-specific interactions. Recent simulations of force-mediated unfolding of FN domains and sequences analysis of human versus mouse FN suggest that the structural stability of the FN’s central cell-binding domains (FN III9-10) affects its integrin specificity. Through production of FN III9-10 variants with variable stabilities, we obtained ligands that present different specificities for the integrin α5β1 and that can be covalently linked into fibrin matrices. Here, we demonstrate the capacity of α5β1 integrin-specific engagement to influence human mesenchymal stem cell (MSC) behavior in 2D and 3D environments. Our data indicate that α5β1 has an important role in the control of MSC osteogenic differentiation. FN fragments with increased specificity for α5β1 versus αvβ3 results in significantly enhanced osteogenic differentiation of MSCs in 2D and in a clinically relevant 3D fibrin matrix system, although attachment/spreading and proliferation were comparable with that on full-length FN. This work shows how integrin-dependant cellular interactions with the ECM can be engineered to control stem cell fate, within a system appropriate for both 3D cell culture and tissue engineering. PMID:19027948

  5. Superresolution microscopy reveals nanometer-scale reorganization of inhibitory natural killer cell receptors upon activation of NKG2D.

    PubMed

    Pageon, Sophie V; Cordoba, Shaun-Paul; Owen, Dylan M; Rothery, Stephen M; Oszmiana, Anna; Davis, Daniel M

    2013-07-23

    Natural killer (NK) cell responses are regulated by a dynamic equilibrium between activating and inhibitory receptor signals at the immune synapse (or interface) with target cells. Although the organization of receptors at the immune synapse is important for appropriate integration of these signals, there is little understanding of this in detail, because research has been hampered by the limited resolution of light microscopy. Through the use of superresolution single-molecule fluorescence microscopy to reveal the organization of the NK cell surface at the single-protein level, we report that the inhibitory receptor KIR2DL1 is organized in nanometer-scale clusters at the surface of human resting NK cells. Nanoclusters of KIR2DL1 became smaller and denser upon engagement of the activating receptor NKG2D, establishing an unexpected crosstalk between activating receptor signals and the positioning of inhibitory receptors. These rearrangements in the nanoscale organization of surface NK cell receptors were dependent on the actin cytoskeleton. Together, these data establish that NK cell activation involves a nanometer-scale reorganization of surface receptors, which in turn affects models for signal integration and thresholds that control NK cell effector functions and NK cell development. PMID:23882121

  6. PPARγ suppresses the proliferation of cardiac myxoma cells through downregulation of MEF2D in a miR-122-dependent manner.

    PubMed

    Qiu, Youzhu; Yang, Jie; Bian, Shizhu; Chen, Guozhu; Yu, Jie

    2016-06-01

    Peroxisome proliferator-activated receptor gamma (PPARγ), a multiple functional transcription factor, has been reported to have anti-tumor effects through inhibition of cells proliferation. However, its effects on cardiac myxoma (CM) cells and the underlying signaling mechanism is unclear. In the present study, we demonstrated that the level of PPARγ is inversely correlated with that of myocyte enhancer factor 2D (MEF2D), a biomarker of CM. We found that activation of PPARγ inhibit MEF2D expression via upregulation of miR-122, which can target the 3'-UTR of MEF2D and inhibit MEF2D expression, by directly binding to the PPRE in the miR-122 promoter region. Functional experiments further showed that miR-122-dependent downregulation of MEF2D by PPARγ suppress the proliferation of CM cells. These results suggest that PPARγ may exert its antiproliferative effects by negatively regulating the MEF2D in CM cells, which through upregulation of miR-122, and PPARγ/miR-122/MEF2D signaling pathway may be a novel target for treatment of CM. PMID:27109478

  7. 2D Computational Fluid Dynamic Modeling of Human Ventricle System Based on Fluid-Solid Interaction and Pulsatile Flow.

    PubMed

    Masoumi, Nafiseh; Framanzad, F; Zamanian, Behnam; Seddighi, A S; Moosavi, M H; Najarian, S; Bastani, Dariush

    2013-01-01

    Many diseases are related to cerebrospinal fluid (CSF) hydrodynamics. Therefore, understanding the hydrodynamics of CSF flow and intracranial pressure is helpful for obtaining deeper knowledge of pathological processes and providing better treatments. Furthermore, engineering a reliable computational method is promising approach for fabricating in vitro models which is essential for inventing generic medicines. A Fluid-Solid Interaction (FSI)model was constructed to simulate CSF flow. An important problem in modeling the CSF flow is the diastolic back flow. In this article, using both rigid and flexible conditions for ventricular system allowed us to evaluate the effect of surrounding brain tissue. Our model assumed an elastic wall for the ventricles and a pulsatile CSF input as its boundary conditions. A comparison of the results and the experimental data was done. The flexible model gave better results because it could reproduce the diastolic back flow mentioned in clinical research studies. The previous rigid models have ignored the brain parenchyma interaction with CSF and so had not reported the back flow during the diastolic time. In this computational fluid dynamic (CFD) analysis, the CSF pressure and flow velocity in different areas were concordant with the experimental data.

  8. 2D Computational Fluid Dynamic Modeling of Human Ventricle System Based on Fluid-Solid Interaction and Pulsatile Flow.

    PubMed

    Masoumi, Nafiseh; Framanzad, F; Zamanian, Behnam; Seddighi, A S; Moosavi, M H; Najarian, S; Bastani, Dariush

    2013-01-01

    Many diseases are related to cerebrospinal fluid (CSF) hydrodynamics. Therefore, understanding the hydrodynamics of CSF flow and intracranial pressure is helpful for obtaining deeper knowledge of pathological processes and providing better treatments. Furthermore, engineering a reliable computational method is promising approach for fabricating in vitro models which is essential for inventing generic medicines. A Fluid-Solid Interaction (FSI)model was constructed to simulate CSF flow. An important problem in modeling the CSF flow is the diastolic back flow. In this article, using both rigid and flexible conditions for ventricular system allowed us to evaluate the effect of surrounding brain tissue. Our model assumed an elastic wall for the ventricles and a pulsatile CSF input as its boundary conditions. A comparison of the results and the experimental data was done. The flexible model gave better results because it could reproduce the diastolic back flow mentioned in clinical research studies. The previous rigid models have ignored the brain parenchyma interaction with CSF and so had not reported the back flow during the diastolic time. In this computational fluid dynamic (CFD) analysis, the CSF pressure and flow velocity in different areas were concordant with the experimental data. PMID:25337330

  9. Synthesizing 2D MoS2 Nanofins on carbon nanospheres as catalyst support for Proton Exchange Membrane Fuel Cells

    PubMed Central

    Hu, Yan; Chua, Daniel H. C.

    2016-01-01

    Highly dense 2D MoS2 fin-like nanostructures on carbon nanospheres were fabricated and formed the main catalyst support structure in the oxygen reduction reaction (ORR) for polymer electrolyte membrane (PEM) fuel cells. These nanofins were observed growing perpendicular to the carbon nanosphere surface in random orientations and high resolution transmission electron microscope confirmed 2D layers. The PEM fuel cell test showed enhanced electrochemical activity with good stability, generating over 8.5 W.mgPt−1 as compared to standard carbon black of 7.4 W.mgPt−1 under normal operating conditions. Electrochemical Impedance Spectroscopy confirmed that the performance improvement is highly due to the excellent water management of the MoS2 lamellar network, which facilitates water retention at low current density and flood prevention at high current density. Reliability test further demonstrated that these nanofins are highly stable in the electrochemical reaction and is an excellent ORR catalyst support. PMID:27302135

  10. Synthesizing 2D MoS2 Nanofins on carbon nanospheres as catalyst support for Proton Exchange Membrane Fuel Cells.

    PubMed

    Hu, Yan; Chua, Daniel H C

    2016-06-15

    Highly dense 2D MoS2 fin-like nanostructures on carbon nanospheres were fabricated and formed the main catalyst support structure in the oxygen reduction reaction (ORR) for polymer electrolyte membrane (PEM) fuel cells. These nanofins were observed growing perpendicular to the carbon nanosphere surface in random orientations and high resolution transmission electron microscope confirmed 2D layers. The PEM fuel cell test showed enhanced electrochemical activity with good stability, generating over 8.5 W.mgPt(-1) as compared to standard carbon black of 7.4 W.mgPt(-1) under normal operating conditions. Electrochemical Impedance Spectroscopy confirmed that the performance improvement is highly due to the excellent water management of the MoS2 lamellar network, which facilitates water retention at low current density and flood prevention at high current density. Reliability test further demonstrated that these nanofins are highly stable in the electrochemical reaction and is an excellent ORR catalyst support.

  11. Synthesizing 2D MoS2 Nanofins on carbon nanospheres as catalyst support for Proton Exchange Membrane Fuel Cells

    NASA Astrophysics Data System (ADS)

    Hu, Yan; Chua, Daniel H. C.

    2016-06-01

    Highly dense 2D MoS2 fin-like nanostructures on carbon nanospheres were fabricated and formed the main catalyst support structure in the oxygen reduction reaction (ORR) for polymer electrolyte membrane (PEM) fuel cells. These nanofins were observed growing perpendicular to the carbon nanosphere surface in random orientations and high resolution transmission electron microscope confirmed 2D layers. The PEM fuel cell test showed enhanced electrochemical activity with good stability, generating over 8.5 W.mgPt-1 as compared to standard carbon black of 7.4 W.mgPt-1 under normal operating conditions. Electrochemical Impedance Spectroscopy confirmed that the performance improvement is highly due to the excellent water management of the MoS2 lamellar network, which facilitates water retention at low current density and flood prevention at high current density. Reliability test further demonstrated that these nanofins are highly stable in the electrochemical reaction and is an excellent ORR catalyst support.

  12. Chimeric NKG2D CAR-Expressing T Cell-Mediated Attack of Human Ovarian Cancer Is Enhanced by Histone Deacetylase Inhibition

    PubMed Central

    Song, De-Gang; Ye, Qunrui; Santoro, Stephen; Fang, Chongyun; Best, Andrew

    2013-01-01

    Abstract NKG2D ligands (NKG2DLs) are widely expressed on ovarian cancers to various degrees, making them attractive targets for immunotherapy. Here, we applied a chimeric antigen receptor (CAR) approach for the targeting of NKG2DLs expressed on human ovarian cancer cells and evaluated the impact of pharmacological upregulation of NKG2DLs on immune recognition. Various NKG2DLs, including MICA/B and ULBP-1, -2, -3, and -4, were expressed at various levels on the surface of all established ovarian cancer cell lines and primary ovarian cancer samples tested. To redirect human T cells against NKG2DLs, an NKG2DL-specific CAR was generated by fusing the extracellular domain of the NKG2D receptor to the 4-1BB costimulatory and CD3-ζ chain signaling domains. In vitro expansion of chimeric NKG2D CAR T cells was delayed compared with untransduced T cells and control CAR T cells; the likely result of fratricide among activated T cells expressing NKG2DLs. However, NKG2D CAR T cells did expand and were selectively enriched during prolonged culture. In coculture, CD4+ and CD8+ NKG2D CAR T cells specifically recognized and killed NKG2DL-expressing ovarian cancer cell lines but not NKG2DL-negative cells. Notably, pretreatment of ovarian cancer cells expressing moderate to low levels of NKG2DLs with the histone deacetylase inhibitor sodium valproate (VPA) upregulated NKG2DL cell surface expression and consequently enhanced their immune recognition by chimeric NKG2D CAR T cells. Our results demonstrate that VPA-induced upregulation of NKG2DL expression enhances the immune recognition of ovarian cancer cells by engineered NKG2D CAR T cells, and rationalizes the use of VPA in combination with NKG2DL-targeted immunotherapy in ovarian cancer. PMID:23297870

  13. A 2D transient numerical model combining heat/mass transport effects in a tubular solid oxide fuel cell

    NASA Astrophysics Data System (ADS)

    Mollayi Barzi, Y.; Ghassemi, M.; Hamedi, M. H.

    The purpose of this study is to present a 2D transient numerical model to predict the dynamic behavior of a tubular SOFC. In this model, the transient conservation equations (momentum, species and energy equations) are solved numerically and electrical and electrochemical outputs are calculated with an equivalent electrical circuit for the cell. The developed model determines the cell electrical and thermal responses to the variation of load current. Also it predicts the local EMF, state variables (pressure, temperature and species concentration) and cell performance for different cell load currents. Using this comprehensive model the dynamic behavior of Tubular SOFC is studied. First an initial steady state operating condition is set for the SOFC model and then the time response of the fuel cell to changes of some interested input parameters (like electrical load) is analyzed. The simulation starts when the cell is at the steady state in a specific output load. When the load step change takes place, the solution continues to reach to the new steady state condition. Then the cell transient behavior is analyzed. The results show that when the load current is stepped up, the output voltage decreases to a new steady state voltage in about 67 min.

  14. Applicability of Modified Drift Flux Model for Bubbly Flow in 2-D/3-D Rectangular Box With Various Kinds of Obstacles

    SciTech Connect

    Tatsuya Matsumoto; Akihiro Uchibori; Ryo Akasaka; Toshinori Seki; Shyuji Kaminishi; Koji Morita; Kenji Fukuda

    2002-07-01

    In order to develop analytical tools for the analyses of multi dimensional two-phase flow in channels with obstacles, the modified drift flux model has been applied. Numerical simulations of multi dimensional gas-liquid two-phase flow in a channel, with some kinds of obstacles inserted to simulate a simple sub-channel in the fuel bundle, were carried out. Analytical results were compared with experiments, to show the validity of the modified drift flux model. Experiments were carried out with using an apparatus of 2-D/3-D rectangular box with a perforated plate or a horizontal plate with slit hole or a vertical rod inserted. Nitrogen gas-water adiabatic two phase flow was circulated in the box. The apparatus was made of acrylic resin plates and be able to make the flow inside visualized. Two-phase flow pattern were recorded with a high-speed video camera and the mass flow rate of nitrogen gas was measured with a digital gas-mass flow meter. Comparisons between the experimental results and the numerical ones showed good agreements, thus it was verified the model would be applied for predicting flows in more complex geometry with obstacles. (authors)

  15. A direct sensitivity comparison between flow-modulated comprehensive 2D and 1D GC in untargeted and targeted MS-based experiments.

    PubMed

    Tranchida, Peter Q; Franchina, Flavio A; Zoccali, Mariosimone; Bonaccorsi, Ivana; Cacciola, Francesco; Mondello, Luigi

    2013-09-01

    The present contribution is focused on the measurement of the analytical sensitivity attained in untargeted/targeted MS/MS experiments, performed using flow-modulator comprehensive 2D and 1D GC. The comprehensive 2D experiment was performed by diverting part of the high flow (circa 80%) to flush the accumulation loop (about 28 mL/min) to waste, to reduce the gas flow entering the ion source. 1D analyses were performed through: (i) unmodulated and (ii) single column applications. An equivalent temperature program was applied in the modulated and unmodulated analyses, while a faster one was employed in the single column one. In all application types, the (same) triple quadrupole instrument was operated in the full-scan and multiple reaction monitoring modes. A genuine sweet orange oil and the same sample spiked with 20 phytosanitary compounds were employed to reach the research objective. The results highlight the problems related to the flow modulation-MS combination. Specifically, it was found that sensitivity was on average three to four times higher in unmodulated and optimized single-column applications. PMID:23868497

  16. Resistance to Cytarabine Induces the Up-regulation of NKG2D Ligands and Enhances Natural Killer Cell Lysis of Leukemic Cells1

    PubMed Central

    Ogbomo, Henry; Michaelis, Martin; Klassert, Denise; Doerr, Hans Wilhelm; Cinatl, Jindrich

    2008-01-01

    Prolonged treatment of leukemic cells with chemotherapeutic agents frequently results in development of drug resistance. Moreover, selection of drug-resistant cell populations may be associated with changes in malignant properties such as proliferation rate, invasiveness, and immunogenicity. In the present study, the sensitivity of cytarabine (1-β-d-arabinofuranosylcytosine, araC)-resistant and parental human leukemic cell lines (T-lymphoid H9 and acute T-lymphoblastic leukemia Molt-4) to natural killer (NK) cell-mediated killing was investigated. The results obtained demonstrate that araC-resistant H9 and Molt-4 (H9rARAC100 and Molt-4rARAC100) cell lines are more sensitive to NK cell-mediated lysis than their respective parental cell lines. This increased sensitivity was associated with a higher surface expression of ligands for the NK cell-activating receptor NKG2D, notably UL16 binding protein-2 (ULBP-2) and ULBP-3 in H9rARAC100 and Molt-4rARAC100 cell lines. Blocking ULBP-2 and ULBP-3 or NKG2D with monoclonal antibody completely abrogated NK cell lysis. Constitutive phosphorylated extracellular signal-regulated kinase (ERK) but not pAKT was higher in araC-resistant cells than in parental cell lines. Inhibition of ERK using ERK inhibitor PD98059 decreased both ULBP-2/ULBP-3 expression and NK cell cytotoxicity. Furthermore, overexpression of constitutively active ERK in H9 parental cells resulted in increased ULBP-2/ULBP-3 expression and enhanced NK cell lysis. These results demonstrate that increased sensitivity of araC-resistant leukemic cells to NK cell lysis is caused by higher NKG2D ligand expression, resulting from more active ERK signaling pathway. PMID:19048119

  17. VFLOW2D - A Vorte-Based Code for Computing Flow Over Elastically Supported Tubes and Tube Arrays

    SciTech Connect

    WOLFE,WALTER P.; STRICKLAND,JAMES H.; HOMICZ,GREGORY F.; GOSSLER,ALBERT A.

    2000-10-11

    A numerical flow model is developed to simulate two-dimensional fluid flow past immersed, elastically supported tube arrays. This work is motivated by the objective of predicting forces and motion associated with both deep-water drilling and production risers in the oil industry. This work has other engineering applications including simulation of flow past tubular heat exchangers or submarine-towed sensor arrays and the flow about parachute ribbons. In the present work, a vortex method is used for solving the unsteady flow field. This method demonstrates inherent advantages over more conventional grid-based computational fluid dynamics. The vortex method is non-iterative, does not require artificial viscosity for stability, displays minimal numerical diffusion, can easily treat moving boundaries, and allows a greatly reduced computational domain since vorticity occupies only a small fraction of the fluid volume. A gridless approach is used in the flow sufficiently distant from surfaces. A Lagrangian remap scheme is used near surfaces to calculate diffusion and convection of vorticity. A fast multipole technique is utilized for efficient calculation of velocity from the vorticity field. The ability of the method to correctly predict lift and drag forces on simple stationary geometries over a broad range of Reynolds numbers is presented.

  18. Transcriptional profiles of valvular interstitial cells cultured on tissue culture polystyrene, on 2D hydrogels, or within 3D hydrogels

    PubMed Central

    Mabry, Kelly M.; Payne, Samuel Z.; Anseth, Kristi S.

    2015-01-01

    Valvular interstitial cells (VICs) actively maintain and repair heart valve tissue; however, persistent activation of VICs to a myofibroblast phenotype can lead to aortic stenosis (Chen and Simmons, 2011) [1]. To better understand and quantify how microenvironmental cues influence VIC phenotype, we compared expression profiles of VICs cultured on/in poly(ethylene glycol) (PEG) gels to those cultured on tissue culture polystyrene (TCPS), as well as fresh isolates. Here, we present both the raw and processed microarray data from these culture conditions. Interpretation of this data can be found in a research article entitled “Microarray analyses to quantify advantages of 2D and 3D hydrogel culture systems in maintaining the native valvular interstitial cell phenotype” (Mabry et al., 2015) [2]. PMID:26702427

  19. A 2D Particle in Cell model for ion extraction and focusing in electrostatic accelerators.

    PubMed

    Veltri, P; Cavenago, M; Serianni, G

    2014-02-01

    Negative ions are fundamental to produce intense and high energy neutral beams used to heat the plasma in fusion devices. The processes regulating the ion extraction involve the formation of a sheath on a scale comparable to the Debye length of the plasma. On the other hand, the ion acceleration as a beam is obtained on distances greater than λD. The paper presents a model for both the phases of ion extraction and acceleration of the ions and its implementation in a numerical code. The space charge of particles is deposited following usual Particle in Cell codes technique, while the field is solved with finite element methods. Some hypotheses on the beam plasma transition are described, allowing to model both regions at the same time. The code was tested with the geometry of the NIO1 negative ions source, and the results are compared with existing ray tracing codes and discussed.

  20. A 2D Particle in Cell model for ion extraction and focusing in electrostatic accelerators

    SciTech Connect

    Veltri, P. Serianni, G.; Cavenago, M.

    2014-02-15

    Negative ions are fundamental to produce intense and high energy neutral beams used to heat the plasma in fusion devices. The processes regulating the ion extraction involve the formation of a sheath on a scale comparable to the Debye length of the plasma. On the other hand, the ion acceleration as a beam is obtained on distances greater than λ{sub D}. The paper presents a model for both the phases of ion extraction and acceleration of the ions and its implementation in a numerical code. The space charge of particles is deposited following usual Particle in Cell codes technique, while the field is solved with finite element methods. Some hypotheses on the beam plasma transition are described, allowing to model both regions at the same time. The code was tested with the geometry of the NIO1 negative ions source, and the results are compared with existing ray tracing codes and discussed.

  1. A 2D Particle in Cell model for ion extraction and focusing in electrostatic accelerators

    NASA Astrophysics Data System (ADS)

    Veltri, P.; Cavenago, M.; Serianni, G.

    2014-02-01

    Negative ions are fundamental to produce intense and high energy neutral beams used to heat the plasma in fusion devices. The processes regulating the ion extraction involve the formation of a sheath on a scale comparable to the Debye length of the plasma. On the other hand, the ion acceleration as a beam is obtained on distances greater than λD. The paper presents a model for both the phases of ion extraction and acceleration of the ions and its implementation in a numerical code. The space charge of particles is deposited following usual Particle in Cell codes technique, while the field is solved with finite element methods. Some hypotheses on the beam plasma transition are described, allowing to model both regions at the same time. The code was tested with the geometry of the NIO1 negative ions source, and the results are compared with existing ray tracing codes and discussed.

  2. Seeing through the trick of cancer cells via 2D gels.

    PubMed

    Mao, Lei

    2013-12-01

    The advancement of modern therapy concepts has dramatically extended the postsurvival rates of patients with malignant gastric cancer. However, a remaining setback is the drug resistance of recurrent cancer, which casts a dark shadow over disease prognosis. The original work of Klein et al. [Proteomics Clin. Appl. 2013, 7, 813-824] has outlined a rational experimental approach to decipher the mechanistic pathway of cancer drug resistance by proteomic approach. They used gel-based comparative proteomics to analyze the nuclear proteome of a human gastric cancer cell line (AGS) with and without inactivation of hypoxia-inducible factor 1 (HIF-1), a transcription factor and master regulator of hypoxia adaptation. Using the classical 2DE-MS approach, these researchers observed 163 HIF-1 responsive proteins, among which over half of them could be confidently identified by MS. From this large dataset, the authors proposed an enhanced nuclear translocation of some proteasomal proteins upon inactivation of HIF-1. Overall, this work appropriately used proteomics as a hypothesis-free, top-down approach to dissect imperative clinical problems.

  3. 2D ratiometric fluorescent pH sensor for tracking of cells proliferation and metabolism.

    PubMed

    Ma, Jun; Ding, Changqin; Zhou, Jie; Tian, Yang

    2015-08-15

    Extracellular pH plays a vital role no matter in physiological or pathological studies. In this work, a hydrogel, CD@Nile-FITC@Gel (Gel sensor), entrapping the ratiometric fluorescent probe CD@Nile-FITC was developed. The Gel sensor was successfully used for real-time extracellular pH monitoring. In the case of CD@Nile-FITC, pH-sensitive fluorescent dye fluorescein isothiocyanate (FITC) was chosen as the response signal for H(+) and Nile blue chloride (Nile) as the reference signal. The developed fluorescent probe exhibited high selectivity for pH over other metal ions and amino acids. Meanwhile, the carbon-dots-based inorganic-organic probe demonstrated excellent photostability against long-term light illumination. In order to study the extracellular pH change in processes of cell proliferation and metabolism, CD@Nile-FITC probe was entrapped in sodium alginate gel and consequently formed CD@Nile-FITC@Gel. The MTT assay showed low cytotoxicity of the Gel and the pH titration indicated that it could monitor the pH fluctuations linearly and rapidly within the pH range of 6.0-9.0, which is valuable for physiological pH determination. As expected, the real-time bioimaging of the probe was successfully achieved.

  4. Identification of the wave speed and the second viscosity in cavitating flow with 2D RANS computations - Part II

    NASA Astrophysics Data System (ADS)

    Alligné, S.; Decaix, J.; Nicolet, C.; Avellan, F.; Münch, C.

    2015-12-01

    The 1D modelling of cavitation vortex rope dynamics in Francis turbine draft tube is decisive for prediction of pressure fluctuations in the system. However, models are defined with parameters which values must be quantified either experimentally or numerically. In this paper a methodology based on CFD simulations is setup to identify these parameters by exciting the flow through outlet boundary condition. A simplified test case is considered to assess if 1D cavitation model parameters can be identified from CFD simulations. It is shown that a low wave speed and a second viscosity due to the cavitating flow can be identified.

  5. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice.

    PubMed

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated.

  6. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice

    NASA Astrophysics Data System (ADS)

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, Yongkeun

    2016-09-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated.

  7. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice

    PubMed Central

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated. PMID:27605489

  8. Holographic intravital microscopy for 2-D and 3-D imaging intact circulating blood cells in microcapillaries of live mice.

    PubMed

    Kim, Kyoohyun; Choe, Kibaek; Park, Inwon; Kim, Pilhan; Park, YongKeun

    2016-01-01

    Intravital microscopy is an essential tool that reveals behaviours of live cells under conditions close to natural physiological states. So far, although various approaches for imaging cells in vivo have been proposed, most require the use of labelling and also provide only qualitative imaging information. Holographic imaging approach based on measuring the refractive index distributions of cells, however, circumvent these problems and offer quantitative and label-free imaging capability. Here, we demonstrate in vivo two- and three-dimensional holographic imaging of circulating blood cells in intact microcapillaries of live mice. The measured refractive index distributions of blood cells provide morphological and biochemical properties including three-dimensional cell shape, haemoglobin concentration, and haemoglobin contents at the individual cell level. With the present method, alterations in blood flow dynamics in live healthy and sepsis-model mice were also investigated. PMID:27605489

  9. Testing the continuum mu(I) rheology for 2D granular flows on avalanches and collapse of columns

    NASA Astrophysics Data System (ADS)

    Lagrée, Pierre-Yves; Staron, Lydie; Popinet, Stéphane

    2010-11-01

    There is a large amount of experimental work dealing with dry granular flows (such as sand, glass beads, small rocks...) supporting the so called μ(I) rheology. This rheology states that the ratio of the tangential to the normal constraints behaves as a Coulomb like friction depending on the Inertial number (this number is the product of the grain size by the shear of the velocity divided by the square root of pressure divided by the grain density). Hence, we propose the implementation of this non newtonian rheology in a Navier Stokes Solver (the Gerris Flow Solver uses a finite-volume approach with the Volume-of-Fluid (VOF) method to describe variable-density two-phase flows). First we apply it on a steady infinite bi dimensional avalanching granular flow over a constant slope covered by a passive light fluid (it allows for a zero pressure boundary condition at the surface, bypassing an up to now difficulty which was to impose this condition on a unknown moving boundary). The classical analytical solution, known as Bagnold solution, is recovered numerically. Then the rheology is tested on the collapse of granular columns and quantitative comparisons with numerical simulations from Contact Dynamics are done.

  10. Developing a 2D vertical flow and sediment transport model for open channels using the Youngs-VOF method

    NASA Astrophysics Data System (ADS)

    Zhao, Dongmiao; Tang, Jun; Wu, Xiuguang; Lin, Changning; Liu, Lijun; Chen, Jian

    2016-05-01

    A 2D vertical (2DV) numerical model, without σ-coordinate transformation in the vertical direction, is developed for the simulation of fl ow and sediment transport in open channels. In the model, time-averaged Reynolds equations are closed by the k-ɛ nonlinear turbulence model. The modifi ed Youngs-VOF method is introduced to capture free surface dynamics, and the free surface slope is simulated using the ELVIRA method. Based on the power-law scheme, the k-ɛ model and the suspended-load transport model are solved numerically with an implicit scheme applied in the vertical plane and an explicit scheme applied in the horizontal plane. Bedload transport is modeled using the Euler-WENO scheme, and the grid-closing skill is adopted to deal with the moving channel bed boundary. Verifi cation of the model using laboratory data shows that the model is able to adequately simulate fl ow and sediment transport in open channels, and is a good starting point for the study of sediment transport dynamics in strong nonlinear fl ow scenarios.

  11. In situ cosmogenic radiocarbon production and 2-D ice flow line modeling for an Antarctic blue ice area

    NASA Astrophysics Data System (ADS)

    Buizert, Christo; Petrenko, Vasilii V.; Kavanaugh, Jeffrey L.; Cuffey, Kurt M.; Lifton, Nathaniel A.; Brook, Edward J.; Severinghaus, Jeffrey P.

    2012-06-01

    Radiocarbon measurements at ice margin sites and blue ice areas can potentially be used for ice dating, ablation rate estimates and paleoclimatic reconstructions. Part of the measured signal comes from in situ cosmogenic 14C production in ice, and this component must be well understood before useful information can be extracted from 14C data. We combine cosmic ray scaling and production estimates with a two-dimensional ice flow line model to study cosmogenic 14C production at Taylor Glacier, Antarctica. We find (1) that 14C production through thermal neutron capture by nitrogen in air bubbles is negligible; (2) that including ice flow patterns caused by basal topography can lead to a surface 14C activity that differs by up to 25% from the activity calculated using an ablation-only approximation, which is used in all prior work; and (3) that at high ablation margin sites, solar modulation of the cosmic ray flux may change the strength of the dominant spallogenic production by up to 10%. As part of this effort we model two-dimensional ice flow along the central flow line of Taylor Glacier. We present two methods for parameterizing vertical strain rates, and assess which method is more reliable for Taylor Glacier. Finally, we present a sensitivity study from which we conclude that uncertainties in published cosmogenic production rates are the largest source of potential error. The results presented here can inform ongoing and future 14C and ice flow studies at ice margin sites, including important paleoclimatic applications such as the reconstruction of paleoatmospheric 14C content of methane.

  12. Section 1. Simulation of surface-water integrated flow and transport in two-dimensions: SWIFT2D user's manual

    USGS Publications Warehouse

    Schaffranek, Raymond W.

    2004-01-01

    A numerical model for simulation of surface-water integrated flow and transport in two (horizontal-space) dimensions is documented. The model solves vertically integrated forms of the equations of mass and momentum conservation and solute transport equations for heat, salt, and constituent fluxes. An equation of state for salt balance directly couples solution of the hydrodynamic and transport equations to account for the horizontal density gradient effects of salt concentrations on flow. The model can be used to simulate the hydrodynamics, transport, and water quality of well-mixed bodies of water, such as estuaries, coastal seas, harbors, lakes, rivers, and inland waterways. The finite-difference model can be applied to geographical areas bounded by any combination of closed land or open water boundaries. The simulation program accounts for sources of internal discharges (such as tributary rivers or hydraulic outfalls), tidal flats, islands, dams, and movable flow barriers or sluices. Water-quality computations can treat reactive and (or) conservative constituents simultaneously. Input requirements include bathymetric and topographic data defining land-surface elevations, time-varying water level or flow conditions at open boundaries, and hydraulic coefficients. Optional input includes the geometry of hydraulic barriers and constituent concentrations at open boundaries. Time-dependent water level, flow, and constituent-concentration data are required for model calibration and verification. Model output consists of printed reports and digital files of numerical results in forms suitable for postprocessing by graphical software programs and (or) scientific visualization packages. The model is compatible with most mainframe, workstation, mini- and micro-computer operating systems and FORTRAN compilers. This report defines the mathematical formulation and computational features of the model, explains the solution technique and related model constraints, describes the

  13. Bifurcation structure of the special class of nonstationary regimes emerging in the 2D inertially coupled, unit-cell model: Analytical study

    NASA Astrophysics Data System (ADS)

    Vorotnikov, K.; Starosvetsky, Y.

    2016-09-01

    Present work is devoted to the analytical investigation of the bifurcation structure of special class of nonstationary low-energy regimes emerging in the locally resonant unit-cell model. System under consideration comprises an outer mass with internal rotator and subject to the 2D, nonlinear local potential. These regimes are characterized by the slow, purely rotational motion of the rotator synchronized with the periodic energy beats between the axial and the lateral vibrations of the outer element. Thus the angular speed of the rotator and the beating frequency of the outer element satisfy the 1:2 resonance condition. In the present study these regimes are referred to as regimes of synchronous nonlinear beats (RSNB). Using the regular muti-scale analysis in the limit of low energy excitation we derive the slow-flow model. To showcase the evolution of RSNBs we used the special Poincaré map technique applied on the slow-flow model. Results of the Poincaré sections unveiled some interesting local bifurcations undergone by these regimes. Further analysis of the slow-flow model enabled us to describe the RSNBs analytically as well as exposed their entire bifurcation structure. The bifurcation analysis has shown the coexistence of several branches of RSNBs corresponding to the regimes of weak and strong, two-dimensional, recurrent energy channeling. We substantiate the results of the analytical study with numerical simulations of the full model and find them to be in the very good agreement.

  14. NKG2D is a Key Receptor for Recognition of Bladder Cancer Cells by IL-2-Activated NK Cells and BCG Promotes NK Cell Activation

    PubMed Central

    García-Cuesta, Eva María; López-Cobo, Sheila; Álvarez-Maestro, Mario; Esteso, Gloria; Romera-Cárdenas, Gema; Rey, Mercedes; Cassady-Cain, Robin L.; Linares, Ana; Valés-Gómez, Alejandro; Reyburn, Hugh Thomson; Martínez-Piñeiro, Luis; Valés-Gómez, Mar

    2015-01-01

    Intravesical instillation of bacillus Calmette–Guérin (BCG) is used to treat superficial bladder cancer, either papillary tumors (after transurethral resection) or high-grade flat carcinomas (carcinoma in situ), reducing recurrence in about 70% of patients. Initially, BCG was proposed to work through an inflammatory response, mediated by phagocytic uptake of mycobacterial antigens and cytokine release. More recently, other immune effectors such as monocytes, natural killer (NK), and NKT cells have been suggested to play a role in this immune response. Here, we provide a comprehensive study of multiple bladder cancer cell lines as putative targets for immune cells and evaluated their recognition by NK cells in the presence and absence of BCG. We describe that different bladder cancer cells can express multiple activating and inhibitory ligands for NK cells. Recognition of bladder cancer cells depended mainly on NKG2D, with a contribution from NKp46. Surprisingly, exposure to BCG did not affect the immune phenotype of bladder cells nor increased NK cell recognition of purified IL-2-activated cell lines. However, NK cells were activated efficiently when BCG was included in mixed lymphocyte cultures, suggesting that NK activation after mycobacteria treatment requires the collaboration of various immune cells. We also analyzed the percentage of NK cells in peripheral blood of a cohort of bladder cancer patients treated with BCG. The total numbers of NK cells did not vary during treatment, indicating that a more detailed study of NK cell activation in the tumor site will be required to evaluate the response in each patient. PMID:26106390

  15. Development of drug loaded nanoparticles for tumor targeting. Part 1: synthesis, characterization, and biological evaluation in 2D cell cultures

    PubMed Central

    El-Dakdouki, Mohammad H.; Puré, Ellen; Huang, Xuefei

    2013-01-01

    Nanoparticles (NPs) are being extensively studied as carriers for drug delivery, but they often have limited penetration inside tumor. We envision that by targeting an endocytic receptor on cell surface, the uptake of NPs can be significantly enhanced through receptor mediated endocytosis. In addition, if the receptor is recycled to cell surface, the NP cargo can be transported out of the cells, which are then taken up by neighboring cells thus enhancing solid tumor penetration. To validate our hypothesis, in the first of two articles, we report the synthesis of doxorubicin (DOX)-loaded, hyaluronan (HA) coated silica nanoparticles (SNP) containing a highly fluorescent core to target CD44, a receptor expressed on cancer cell surface. HA was conjugated onto amine-functionalized SNPs prepared through an oil/water microemulsion method. The immobilization of the cytotoxic drug DOX was achieved through an acid sensitive hydrazone linkage. The NPs were fully characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS), zeta potential measurements, thermal gravimetric analysis (TGA), UV-vis absorbance, and nuclear magnetic resonance (NMR). Initial biological evaluation experiments demonstrated that compared to ligand-free SNPs, the uptake of HA-SNP by the CD44-expressing SKOV-3 ovarian cancer cells was significantly enhanced when evaluated in the 2D monolayer cell culture. Mechanistic studies suggested that cellular uptake of HA-SNP was mainly through CD44 mediated endocytosis. HA-SNPs with DOX immobilized were endocytosed efficiently by the SKOV-3 cells as well. The enhanced tumor penetration and drug delivery properties of HA-SNP will be evaluated in 3D tumor models in the subsequent paper. PMID:23529646

  16. ICEG2D (v2.0) - An Integrated Software Package for Automated Prediction of Flow Fields for Single-Element Airfoils With Ice Accretion

    NASA Technical Reports Server (NTRS)

    Thompson David S.; Soni, Bharat K.

    2001-01-01

    An integrated geometry/grid/simulation software package, ICEG2D, is being developed to automate computational fluid dynamics (CFD) simulations for single- and multi-element airfoils with ice accretions. The current version, ICEG213 (v2.0), was designed to automatically perform four primary functions: (1) generate a grid-ready surface definition based on the geometrical characteristics of the iced airfoil surface, (2) generate high-quality structured and generalized grids starting from a defined surface definition, (3) generate the input and restart files needed to run the structured grid CFD solver NPARC or the generalized grid CFD solver HYBFL2D, and (4) using the flow solutions, generate solution-adaptive grids. ICEG2D (v2.0) can be operated in either a batch mode using a script file or in an interactive mode by entering directives from a command line within a Unix shell. This report summarizes activities completed in the first two years of a three-year research and development program to address automation issues related to CFD simulations for airfoils with ice accretions. As well as describing the technology employed in the software, this document serves as a users manual providing installation and operating instructions. An evaluation of the software is also presented.

  17. Experimental results for film cooling in 2-D supersonic flow including coolant delivery pressure, geometry, and incident shock effects

    NASA Technical Reports Server (NTRS)

    Olsen, George C.; Nowak, Robert J.; Holden, Michael S.; Baker, N. R.

    1990-01-01

    An experimental program was conducted to establish some design parameters important to a supersonic film cooling system in a scramjet engine. A simple non-combusting two-dimensional flow configuration was used to isolate the film cooling phenomena. Parameters investigated include coolant delivery pressure, slot height and lip thickness, and incident shock location and strength. Design guidelines for use in engineering and trade studies are presented.

  18. Multi-GPU unsteady 2D flow simulation coupled with a state-to-state chemical kinetics

    NASA Astrophysics Data System (ADS)

    Tuttafesta, Michele; Pascazio, Giuseppe; Colonna, Gianpiero

    2016-10-01

    In this work we are presenting a GPU version of a CFD code for high enthalpy reacting flow, using the state-to-state approach. In supersonic and hypersonic flows, thermal and chemical non-equilibrium is one of the fundamental aspects that must be taken into account for the accurate characterization of the plasma and state-to-state kinetics is the most accurate approach used for this kind of problems. This model consists in writing a continuity equation for the population of each vibrational level of the molecules in the mixture, determining at the same time the species densities and the distribution of the population in internal levels. An explicit scheme is employed here to integrate the governing equations, so as to exploit the GPU structure and obtain an efficient algorithm. The best performances are obtained for reacting flows in state-to-state approach, reaching speedups of the order of 100, thanks to the use of an operator splitting scheme for the kinetics equations.

  19. Cytokine-induced killer cells efficiently kill stem-like cancer cells of nasopharyngeal carcinoma via the NKG2D-ligands recognition.

    PubMed

    Wei, Fang; Rong, Xiao-Xiang; Xie, Rao-Ying; Jia, Li-Ting; Wang, Hui-Yan; Qin, Yu-Juan; Chen, Lin; Shen, Hong-Fen; Lin, Xiao-Lin; Yang, Jie; Yang, Sheng; Hao, Wei-Chao; Chen, Yan; Xiao, Sheng-Jun; Zhou, Hui-Rong; Lin, Tao-Yan; Chen, Yu-Shuang; Sun, Yan; Yao, Kai-Tai; Xiao, Dong

    2015-10-27

    Cancer stem cells (CSCs) are considered to be the root cause for cancer treatment failure. Thus, there remains an urgent need for more potent and safer therapies against CSCs for curing cancer. In this study, the antitumor activity of cytokine-induced killer (CIK) cells against putative CSCs of nasopharyngeal carcinoma (NPC) was fully evaluated in vitro and in vivo. To visualize putative CSCs in vitro by fluorescence imaging, and image and quantify putative CSCs in tumor xenograft-bearing mice by in vivo bioluminescence imaging, NPC cells were engineered with CSC detector vector encoding GFP and luciferase (Luc) under control of Nanog promoter. Our study reported in vitro intense tumor-killing activity of CIK cells against putative CSCs of NPC, as revealed by percentage analysis of side population cells, tumorsphere formation assay and Nanog-promoter-GFP-Luc reporter gene strategy plus time-lapse recording. Additionally, time-lapse imaging firstly illustrated that GFP-labeled or PKH26-labeled putative CSCs or tumorspheres were usually attacked simultaneously by many CIK cells and finally killed by CIK cells, suggesting the necessity of achieving sufficient effector-to-target ratios. We firstly confirmed that NKG2D blockade by anti-NKG2D antibody significantly but partially abrogated CIK cell-mediated cytolysis against putative CSCs. More importantly, intravenous infusion of CIK cells significantly delayed tumor growth in NOD/SCID mice, accompanied by a remarkable reduction in putative CSC number monitored by whole-body bioluminescence imaging. Taken together, our findings suggest that CIK cells demonstrate the intense tumor-killing activity against putative CSCs of NPC, at least in part, by NKG2D-ligands recognition. These results indicate that CIK cell-based therapeutic strategy against CSCs presents a promising and safe approach for cancer treatment. PMID:26418951

  20. RAE-1 ligands for the NKG2D receptor are regulated by E2F transcription factors, which control cell cycle entry

    PubMed Central

    Jung, Heiyoun; Hsiung, Benjamin; Pestal, Kathleen; Procyk, Emily

    2012-01-01

    The NKG2D stimulatory receptor expressed by natural killer cells and T cell subsets recognizes cell surface ligands that are induced on transformed and infected cells and facilitate immune rejection of tumor cells. We demonstrate that expression of retinoic acid early inducible gene 1 (RAE-1) family NKG2D ligands in cancer cell lines and proliferating normal cells is coupled directly to cell cycle regulation. Raet1 genes are directly transcriptionally activated by E2F family transcription factors, which play a central role in regulating cell cycle entry. Induction of RAE-1 occurred in primary cell cultures, embryonic brain cells in vivo, and cells in healing skin wounds and, accordingly, wound healing was delayed in mice lacking NKG2D. Transcriptional activation by E2Fs is likely coordinated with posttranscriptional regulation by other stress responses. These findings suggest that cellular proliferation, as occurs in cancer cells but also other pathological conditions, is a key signal tied to immune reactions mediated by NKG2D-bearing lymphocytes. PMID:23166357

  1. Instantaneous 2D Velocity and Temperature Measurements in High Speed Flows Based on Spectrally Resolved Molecular Rayleigh Scattering

    NASA Technical Reports Server (NTRS)

    Seasholtz, Richard G.

    1995-01-01

    A Rayleigh scattering diagnostic for high speed flows is described for the simultaneous, instantaneous measurement of gas temperature and velocity at a number (up to about one hundred) of locations in a plane illuminated by an injection-seeded, frequency doubled Nd:YAG laser. Molecular Rayleigh scattered light is collected and passed through a planar mirror Fabry-Perot interferometer. The resulting image is analyzed to determine the gas temperature and bulk velocity at each of the regions. The Cramer Rao lower bound for measurement uncertainty is calculated. Experimental data is presented for a free jet and for preliminary measurements in the Lewis 4 inch by 10 inch supersonic wind tunnel.

  2. Human Lung Cancer Cells Grown in an Ex Vivo 3D Lung Model Produce Matrix Metalloproteinases Not Produced in 2D Culture

    PubMed Central

    Mishra, Dhruva K.; Sakamoto, Jason H.; Thrall, Michael J.; Baird, Brandi N.; Blackmon, Shanda H.; Ferrari, Mauro; Kurie, Jonathan M.; Kim, Min P.

    2012-01-01

    We compared the growth of human lung cancer cells in an ex vivo three-dimensional (3D) lung model and 2D culture to determine which better mimics lung cancer growth in patients. A549 cells were grown in an ex vivo 3D lung model and in 2D culture for 15 days. We measured the size and formation of tumor nodules and counted the cells after 15 days. We also stained the tissue/cells for Ki-67, and Caspase-3. We measured matrix metalloproteinase (MMP) levels in the conditioned media and in blood plasma from patients with adenocarcinoma of the lung. Organized tumor nodules with intact vascular space formed in the ex vivo 3D lung model but not in 2D culture. Proliferation and apoptosis were greater in the ex vivo 3D lung model compared to the 2D culture. After 15 days, there were significantly more cells in the 2D culture than the 3D model. MMP-1, MMP-9, and MMP-10 production were significantly greater in the ex vivo 3D lung model. There was no production of MMP-9 in the 2D culture. The patient samples contained MMP-1, MMP-2, MMP-9, and MMP-10. The human lung cancer cells grown on ex vivo 3D model form perfusable nodules that grow over time. It also produced MMPs that were not produced in 2D culture but seen in human lung cancer patients. The ex vivo 3D lung model may more closely mimic the biology of human lung cancer development than the 2D culture. PMID:23028922

  3. Fringe-controlled biodegradation under dynamic conditions: Quasi 2-D flow-through experiments and reactive-transport modeling

    NASA Astrophysics Data System (ADS)

    Eckert, Dominik; Kürzinger, Petra; Bauer, Robert; Griebler, Christian; Cirpka, Olaf A.

    2015-01-01

    Biodegradation in contaminated aquifers has been shown to be most pronounced at the fringe of contaminant plumes, where mixing of contaminated water and ambient groundwater, containing dissolved electron acceptors, stimulates microbial activity. While physical mixing of contaminant and electron acceptor by transverse dispersion has been shown to be the major bottleneck for biodegradation in steady-state plumes, so far little is known on the effect of flow and transport dynamics (caused, e.g., by a seasonally fluctuating groundwater table) on biodegradation in these systems. Towards this end we performed experiments in quasi-two-dimensional flow-through microcosms on aerobic toluene degradation by Pseudomonas putida F1. Plume dynamics were simulated by vertical alteration of the toluene plume position and experimental results were analyzed by reactive-transport modeling. We found that, even after disappearance of the toluene plume for two weeks, the majority of microorganisms stayed attached to the sediment and regained their full biodegradation potential within two days after reappearance of the toluene plume. Our results underline that besides microbial growth, also maintenance and dormancy are important processes that affect biodegradation performance under transient environmental conditions and therefore deserve increased consideration in future reactive-transport modeling.

  4. 2D dry granular free-surface flow over complex topography with obstacles. Part I: experimental study using a consumer-grade RGB-D sensor

    NASA Astrophysics Data System (ADS)

    Caviedes-Voullième, Daniel; Juez, Carmelo; Murillo, Javier; García-Navarro, Pilar

    2014-12-01

    Avalanches, debris flows and other types of gravity-driven granular flows are a common hazard in mountainous regions. These regions often have human settlements in the lower parts of valleys, with human structures dangerously exposed to the destructive effects of these geophysical flows. Therefore a scientific effort has been made to understand, model and simulate geophysical granular flows. In order for computer models and simulations to be of predictive value they need to be validated under controlled, yet nature-like conditions. This work presents an experimental study of granular flow over a simplified mountain slope and valley topography. The experimental facility has a rough bed with very high slope at the upstream end and adverse slope on the downstream end, following a parabolic profile. Obstacles are present in the lower regions. Transient measurements of the moving granular surfaces were taken with a consumer-grade RGB-D sensor, providing transient 2D elevation fields around the obstacles. Three experimental configurations were tested, with semispheres of different diameters and a square dike obstacle. The experimental results are very consistent and repeatable. The quantitative, transient and two-dimensional data for all three experiments constitute excellent benchmarking tests for computational models, such as the one presented in a companion paper.

  5. A novel 1D/2D model for simulating conjugate heat transfer applied to flow boiling in tubes with external fins

    NASA Astrophysics Data System (ADS)

    Ocłoń, Paweł; Łopata, Stanisław; Nowak, Marzena

    2014-09-01

    This study presents a novel, simplified model for the time-efficient simulation of transient conjugate heat transfer in round tubes. The flow domain and the tube wall are modeled in 1D and 2D, respectively and empirical correlations are used to model the flow domain in 1D. The model is particularly useful when dealing with complex physics, such as flow boiling, which is the main focus of this study. The tube wall is assumed to have external fins. The flow is vertical upwards. Note that straightforward computational fluid dynamics (CFD) analysis of conjugate heat transfer in a system of tubes, leads to 3D modeling of fluid and solid domains. Because correlation is used and dimensionality reduced, the model is numerically more stable and computationally more time-efficient compared to the CFD approach. The benefit of the proposed approach is that it can be applied to large systems of tubes as encountered in many practical applications. The modeled equations are discretized in space using the finite volume method, with central differencing for the heat conduction equation in the solid domain, and upwind differencing of the convective term of the enthalpy transport equation in the flow domain. An explicit time discretization with forward differencing was applied to the enthalpy transport equation in the fluid domain. The conduction equation in the solid domain was time discretized using the Crank-Nicholson scheme. The model is applied in different boundary conditions and the predicted boiling patterns and temperature fields are discussed.

  6. A novel 1D/2D model for simulating conjugate heat transfer applied to flow boiling in tubes with external fins

    NASA Astrophysics Data System (ADS)

    Ocłoń, Paweł; Łopata, Stanisław; Nowak, Marzena

    2015-04-01

    This study presents a novel, simplified model for the time-efficient simulation of transient conjugate heat transfer in round tubes. The flow domain and the tube wall are modeled in 1D and 2D, respectively and empirical correlations are used to model the flow domain in 1D. The model is particularly useful when dealing with complex physics, such as flow boiling, which is the main focus of this study. The tube wall is assumed to have external fins. The flow is vertical upwards. Note that straightforward computational fluid dynamics (CFD) analysis of conjugate heat transfer in a system of tubes, leads to 3D modeling of fluid and solid domains. Because correlation is used and dimensionality reduced, the model is numerically more stable and computationally more time-efficient compared to the CFD approach. The benefit of the proposed approach is that it can be applied to large systems of tubes as encountered in many practical applications. The modeled equations are discretized in space using the finite volume method, with central differencing for the heat conduction equation in the solid domain, and upwind differencing of the convective term of the enthalpy transport equation in the flow domain. An explicit time discretization with forward differencing was applied to the enthalpy transport equation in the fluid domain. The conduction equation in the solid domain was time discretized using the Crank-Nicholson scheme. The model is applied in different boundary conditions and the predicted boiling patterns and temperature fields are discussed.

  7. PC2D simulation and optimization of the selective emitter solar cells fabricated by screen printing phosphoric paste method

    NASA Astrophysics Data System (ADS)

    Jia, Xiaojie; Ai, Bin; Deng, Youjun; Xu, Xinxiang; Peng, Hua; Shen, Hui

    2015-08-01

    On the basis of perfect PC2D simulation to the measured current density vs voltage (J-V) curve of the best selective emitter (SE) solar cell fabricated by the CSG Company using the screen printing phosphoric paste method, we systematically investigated the effect of the parameters of gridline, base, selective emitter, back surface field (BSF) layer and surface recombination rate on performance of the SE solar cell. Among these parameters, we identified that the base minority carrier lifetime, the front and back surface recombination rate and the ratio of the sheet-resistance of heavily and lightly doped region are the four largest efficiency-affecting factors. If all the parameters have ideal values, the SE solar cell fabricated on a p-type monocrystalline silicon wafer can even obtain the efficiency of 20.45%. In addition, the simulation also shows that fine gridline combining dense gridline and increasing bus bar number while keeping the lower area ratio can offer the other ways to improve the efficiency.

  8. Parameterization of a numerical 2-D debris flow model with entrainment: a case study of the Faucon catchment, Southern French Alps

    NASA Astrophysics Data System (ADS)

    Hussin, H. Y.; Luna, B. Quan; van Westen, C. J.; Christen, M.; Malet, J.-P.; van Asch, Th. W. J.

    2012-10-01

    The occurrence of debris flows has been recorded for more than a century in the European Alps, accounting for the risk to settlements and other human infrastructure that have led to death, building damage and traffic disruptions. One of the difficulties in the quantitative hazard assessment of debris flows is estimating the run-out behavior, which includes the run-out distance and the related hazard intensities like the height and velocity of a debris flow. In addition, as observed in the French Alps, the process of entrainment of material during the run-out can be 10-50 times in volume with respect to the initially mobilized mass triggered at the source area. The entrainment process is evidently an important factor that can further determine the magnitude and intensity of debris flows. Research on numerical modeling of debris flow entrainment is still ongoing and involves some difficulties. This is partly due to our lack of knowledge of the actual process of the uptake and incorporation of material and due the effect of entrainment on the final behavior of a debris flow. Therefore, it is important to model the effects of this key erosional process on the formation of run-outs and related intensities. In this study we analyzed a debris flow with high entrainment rates that occurred in 2003 at the Faucon catchment in the Barcelonnette Basin (Southern French Alps). The historic event was back-analyzed using the Voellmy rheology and an entrainment model imbedded in the RAMMS 2-D numerical modeling software. A sensitivity analysis of the rheological and entrainment parameters was carried out and the effects of modeling with entrainment on the debris flow run-out, height and velocity were assessed.

  9. Substantial increase in the frequency of circulating CD4+NKG2D+ T cells in patients with cervical intraepithelial neoplasia grade 1

    PubMed Central

    2013-01-01

    Background The NKG2D receptor confers important activating signals to NK cells via ligands expressed during cellular stress and viral infection. This receptor has generated great interest because not only is it expressed on NK cells, but it is also seen in virtually all CD8+ cytotoxic T cells and is classically considered absent in CD4+ T cells. However, recent studies have identified a distinctive population of CD4+ T cells that do express NKG2D, which could represent a particular cytotoxic effector population involved in viral infections and chronic diseases. On the other hand, increased incidence of human papillomavirus-associated lesions in CD4+ T cell-immunocompromised individuals suggests that CD4+ T cells play a key role in controlling the viral infection. Therefore, this study was focused on identifying the frequency of NKG2D-expressing CD4+ T cells in patients with cervical intraepithelial neoplasia (CIN) 1. Additionally, factors influencing CD4+NKG2D+ T cell expansion were also measured. Results Close to 50% of patients with CIN 1 contained at least one of the 37 HPV types detected by our genotyping system. A tendency for increased CD4+ T cells and CD8+ T cells and decreased NK cells was found in CIN 1 patients. The percentage of circulating CD4+ T cells co-expressing the NKG2D receptor significantly increased in women with CIN 1 versus control group. Interestingly, the increase of CD4+NKG2D+ T cells was seen in patients with CIN 1, despite the overall levels of CD4+ T cells did not significantly increase. We also found a significant increase of soluble MICB in CIN 1 patients; however, no correlation with the presence of CD4+NKG2D+ T cells was seen. While TGF-beta was significantly decreased in the group of CIN 1 patients, both TNF-alpha and IL-15 showed a tendency to increase in this group. Conclusions Taken together, our results suggest that the significant increase within the CD4+NKG2D+ T cell population in CIN 1 patients might be the result of a

  10. Selection and Testing of Pressure and Temperature Sensitive Dyes for 2-D Flow Characterization via Synthesized Microbeads

    NASA Astrophysics Data System (ADS)

    Perez, Alex; Zhu, Cun; Xia, Younan; Khalil, Gamal; Dabiri, Dana

    2011-11-01

    Airborne temperature and pressure sensitive microbeads provide a vehicle with which to conduct two-dimensional flow characterization. An array of temperature and pressure sensitive dyes have been synthesized with microbeads (of silica, polystyrene, and polydimethylsiloxane) for this purpose. These microbeads were evaluated based on emission spectra, pressure response (0-760 torr), temperature response (5-45°C), and response time. Work will be presented showing the various combinations of dyes and microbead materials, as well as the testing process and examples of future application. This material is based upon work supported by the National Science Foundation Graduate Research Fellowship under Grant No. #DEG-0718124, as well as National Science Foundation Grant No. NSF/CBET-IDR- 0929864.

  11. Use of Caval Subtraction 2D Phase-Contrast MR Imaging to Measure Total Liver and Hepatic Arterial Blood Flow: Preclinical Validation and Initial Clinical Translation.

    PubMed

    Chouhan, Manil D; Mookerjee, Rajeshwar P; Bainbridge, Alan; Walker-Samuel, Simon; Davies, Nathan; Halligan, Steve; Lythgoe, Mark F; Taylor, Stuart A

    2016-09-01

    Purpose To validate caval subtraction two-dimensional (2D) phase-contrast magnetic resonance (MR) imaging measurements of total liver blood flow (TLBF) and hepatic arterial fraction in an animal model and evaluate consistency and reproducibility in humans. Materials and Methods Approval from the institutional ethical committee for animal care and research ethics was obtained. Fifteen Sprague-Dawley rats underwent 2D phase-contrast MR imaging of the portal vein (PV) and infrahepatic and suprahepatic inferior vena cava (IVC). TLBF and hepatic arterial flow were estimated by subtracting infrahepatic from suprahepatic IVC flow and PV flow from estimated TLBF, respectively. Direct PV transit-time ultrasonography (US) and fluorescent microsphere measurements of hepatic arterial fraction were the standards of reference. Thereafter, consistency of caval subtraction phase-contrast MR imaging-derived TLBF and hepatic arterial flow was assessed in 13 volunteers (mean age, 28.3 years ± 1.4) against directly measured phase-contrast MR imaging PV and proper hepatic arterial inflow; reproducibility was measured after 7 days. Bland-Altman analysis of agreement and coefficient of variation comparisons were undertaken. Results There was good agreement between PV flow measured with phase-contrast MR imaging and that measured with transit-time US (mean difference, -3.5 mL/min/100 g; 95% limits of agreement [LOA], ±61.3 mL/min/100 g). Hepatic arterial fraction obtained with caval subtraction agreed well with those with fluorescent microspheres (mean difference, 4.2%; 95% LOA, ±20.5%). Good consistency was demonstrated between TLBF in humans measured with caval subtraction and direct inflow phase-contrast MR imaging (mean difference, -1.3 mL/min/100 g; 95% LOA, ±23.1 mL/min/100 g). TLBF reproducibility at 7 days was similar between the two methods (95% LOA, ±31.6 mL/min/100 g vs ±29.6 mL/min/100 g). Conclusion Caval subtraction phase-contrast MR imaging is a simple and clinically

  12. 1α,25(OH)2D3 Suppresses the Migration of Ovarian Cancer SKOV-3 Cells through the Inhibition of Epithelial–Mesenchymal Transition

    PubMed Central

    Hou, Yong-Feng; Gao, Si-Hai; Wang, Ping; Zhang, He-Mei; Liu, Li-Zhi; Ye, Meng-Xuan; Zhou, Guang-Ming; Zhang, Zeng-Li; Li, Bing-Yan

    2016-01-01

    Ovarian cancer is the most lethal gynecological malignancy due to its high metastatic ability. Epithelial-mesenchymal transition (EMT) is essential during both follicular rupture and epithelium regeneration. However, it may also accelerate the progression of ovarian carcinomas. Experimental studies have found that 1α,25-dihydroxyvitamin-D3 [1α,25(OH)2D3] can inhibit the proliferation of ovarian cancer cells. In this study, we investigated whether 1α,25(OH)2D3 could inhibit the migration of ovarian cancer cells via regulating EMT. We established a model of transient transforming growth factor-β1(TGF-β1)-induced EMT in human ovarian adenocarcinoma cell line SKOV-3 cells. Results showed that, compared with control, 1α,25(OH)2D3 not only inhibited the migration and the invasion of SKOV-3 cells, but also promoted the acquisition of an epithelial phenotype of SKOV-3 cells treated with TGF-β1. We discovered that 1α,25(OH)2D3 increased the expression of epithelial marker E-cadherin and decreased the level of mesenchymal marker, Vimentin, which was associated with the elevated expression of VDR. Moreover, 1α,25(OH)2D3 reduced the expression level of transcription factors of EMT, such as slug, snail, and β-catenin. These results indicate that 1α,25(OH)2D3 suppresses the migration and invasion of ovarian cancer cells by inhibiting EMT, implying that 1α,25(OH)2D3 might be a potential therapeutic agent for the treatment of ovarian cancer. PMID:27548154

  13. Rising of a single Taylor drop in a stagnant liquid—2D laminar flow and axisymmetry limits

    NASA Astrophysics Data System (ADS)

    Direito, F. J. N.; Campos, J. B. L. M.; Miranda, J. M.

    2016-05-01

    A numerical (computational fluid dynamics (CFD)) study concerning the rise of individual liquid Taylor drops through vertical columns of stagnant heavier liquids is presented in this paper. CFD simulations were performed in Ansys Fluent, using its implementation of volume of fluid method, assuming the flow to be axisymmetric and laminar. Different physical conditions were tested, corresponding to different combinations of relevant dimensionless parameters and the numerical method was validated through experimental data available in the literature. The viscosity ratio between the lighter and the heavier liquid was within the range 0.01-40 and Eötvös number was between 8 and 30. Morton number was within the interval of 2.32 × 10-6-100. Froude number results were compared to data from a literature correlation. The accordance is acceptable for the ranges studied. Velocity profiles in significant regions are reported (drop nose, drop bottom and continuous phase liquid film). The influence of changing one dimensionless parameter alone was assessed. For small and large viscosity ratios, axisymmetric behavior is not a valid assumption.

  14. 2D-DIGE proteomic analysis of mesenchymal stem cell cultured on the elasticity-tunable hydrogels.

    PubMed

    Kuboki, Thasaneeya; Kantawong, Fahsai; Burchmore, Richard; Dalby, Matthew J; Kidoaki, Satoru

    2012-01-01

    The present study focuses on mechanotransduction in mesenchymal stem cells (MSCs) in response to matrix elasticity. By using photocurable gelatinous gels with tunable stiffness, proteomic profiles of MSCs cultured on tissue culture plastic, soft (3 kPa) and stiff (52 kPa) matrices were deciphered using 2-dimensional differential in-gel analysis (2D-DIGE). The DIGE data, tied to immunofluorescence, indicated abundance and organization changes in the cytoskeletonal proteins as well as differential regulation of important signaling-related proteins, stress-responsing proteins and also proteins involved in collagen synthesis. The major CSK proteins including actin, tubulin and vimentin of the cells cultured on the gels were remarkably changed their expressions. Significant down-regulation of α-tubulin and β-actin can be observed on gel samples in comparison to the rigid tissue culture plates. The expression abundance of vimentin appeared to be highest in the MSCs cultured on hard gels. These results suggested that the substrate stiffness significantly affects expression balances in cytoskeletal proteins of MSCs with some implications to cellular tensegrity. PMID:22971925

  15. Antigen Presenting Cells and Stromal Cells Trigger Human Natural Killer Lymphocytes to Autoreactivity: Evidence for the Involvement of Natural Cytotoxicity Receptors (NCR) and NKG2D

    PubMed Central

    Poggi, Alessandro; Zocchi, Maria Raffaella

    2006-01-01

    Human natural killer (NK) lymphocytes should not damage autologous cells due to the engagement of inhibitory receptor superfamily (IRS) members by HLA-I. Nevertheless, NK cells kill self cells expressing low levels or lacking HLA-I, as it may occur during viral infections (missing-self hypothesis). Herein, we show that human NK cells can be activated upon binding with self antigen presenting cells or stromal cells despite the expression of HLA-I. Indeed, NK cells can kill and produce pro-inflammatory and regulating cytokines as IFN-γ, TNF-α and IL10 during interaction with autologous dendritic cells or bone marrow stromal cells or skin fibroblasts. The killing of antigen presenting and stromal cells is dependent on LFA1/ICAM1 interaction. Further, the natural cytotoxicity receptors (NCR) NKp30 and NKp46 are responsible for the delivery of lethal hit to DC, whereas NKG2D activating receptor, the ligand of the MHC-related molecule MIC-A and the UL16 binding protein, is involved in stromal cell killing. These findings indicate that different activating receptors are involved in cell to self cell interaction. Finally, NK cells can revert the veto effect of stromal cells on mixed lymphocyte reaction further supporting the idea that NK cells may alter the interaction between T lymphocytes and microenvironment leading to autoreactivity. PMID:17162374

  16. 2D FTLE in 3D Flows: The accuracy of using two-dimensional data for Lagrangian analysis in a three-dimensional turbulent channel simulation

    NASA Astrophysics Data System (ADS)

    Rockwood, Matthew; Green, Melissa

    2012-11-01

    In experimental, three-dimensional vortex-dominated flows, common particle image velocimetry (PIV) data is often collected in only the plane of interest due to equipment constraints. For flows with significant out of plane velocities or velocity gradients, this can create large discrepancies in Lagrangian analyses that require accurate particle trajectories. A Finite Time Lyapunov Exponent (FTLE) analysis is one such example, and has been shown to be very powerful at examining vortex dynamics and interactions in a variety of aperiodic flows. In this work, FTLE analysis of a turbulent channel simulation was conducted using both full three-dimensional velocity data and modified planar data extracted from the same computational domain. When the out of plane velocity component is neglected the difference in FTLE fields is non-trivial. A quantitative comparison and computation of error is presented for several planes across the width of the channel to determine the efficacy of using 2D analyses on the inherently 3D flows.

  17. Detection of multi-scale secondary flow structures using anisotropic 2D Ricker wavelets in a bent tube model for curved arteries

    NASA Astrophysics Data System (ADS)

    Plesniak, Daniel H.; Bulusu, Kartik V.; Plesniak, Michael W.

    2012-11-01

    Interpretation of complex flow patterns observed in this study of a model curved artery required characterization of multiple, low-circulation secondary flow structures that were observed during the late systolic deceleration and diastolic phases under physiological inflow conditions. Phase-locked, planar vorticity PIV data were acquired at various cross-sectional locations of the 180-degree bent tube model. High circulation, deformed Dean- and Lyne-type vortices were observed during early stages of deceleration, while several smaller scale, highly deformed, low-circulation vortical patterns appeared in the core and near-wall regions during late systolic deceleration and diastolic phases. Due to the multiplicity of vortical scales and shapes, anisotropic 2D Ricker wavelets were used for coherent structure detection in a continuous wavelet transform algorithm (PIVlet 1.2). Our bio-inspired study is geared towards understanding whether optimizing the shape of the wavelet kernel will enable better resolution of several low-circulation, multi-scale secondary flow morphologies and whether new insights into the dynamics of arterial secondary flow structures can accordingly be gained. Supported by the National Science Foundation, Grant No. CBET-0828903 and GW Center for Biomimetics and Bioinspired Engineering (COBRE).

  18. Wind-tunnel experiments of turbulent flow over a surface-mounted 2-D block in a thermally-stratified boundary layer

    NASA Astrophysics Data System (ADS)

    Zhang, Wei; Markfort, Corey; Porté-Agel, Fernando

    2014-11-01

    Turbulent flows over complex surface topography have been of great interest in the atmospheric science and wind engineering communities. The geometry of the topography, surface roughness and temperature characteristics as well as the atmospheric thermal stability play important roles in determining momentum and scalar flux distribution. Studies of turbulent flow over simplified topography models, under neutrally stratified boundary-layer conditions, have provided insights into fluid dynamics. However, atmospheric thermal stability has rarely been considered in laboratory experiments, e.g., wind-tunnel experiments. Series of wind-tunnel experiments of thermally-stratified boundary-layer flow over a surface-mounted 2-D block, in a well-controlled boundary-layer wind tunnel, will be presented. Measurements using high-resolution PIV, x-wire/cold-wire anemometry and surface heat flux sensors were conducted to quantify the turbulent flow properties, including the size of the recirculation zone, coherent vortex structures and the subsequent boundary layer recovery. Results will be shown to address thermal stability effects on momentum and scalar flux distribution in the wake, as well as dominant mechanism of turbulent kinetic energy generation and consumption. The authors gratefully acknowledge funding from the Swiss National Foundation (Grant 200021-132122), the National Science Foundation (Grant ATM-0854766) and NASA (Grant NNG06GE256).

  19. Mesh2d

    2011-12-31

    Mesh2d is a Fortran90 program designed to generate two-dimensional structured grids of the form [x(i),y(i,j)] where [x,y] are grid coordinates identified by indices (i,j). The x(i) coordinates alone can be used to specify a one-dimensional grid. Because the x-coordinates vary only with the i index, a two-dimensional grid is composed in part of straight vertical lines. However, the nominally horizontal y(i,j0) coordinates along index i are permitted to undulate or otherwise vary. Mesh2d also assignsmore » an integer material type to each grid cell, mtyp(i,j), in a user-specified manner. The complete grid is specified through three separate input files defining the x(i), y(i,j), and mtyp(i,j) variations.« less

  20. Modelling river bank erosion using a 2D depth-averaged numerical model of flow and non-cohesive, non-uniform sediment transport

    NASA Astrophysics Data System (ADS)

    El Kadi Abderrezzak, Kamal; Die Moran, Andrés; Tassi, Pablo; Ata, Riadh; Hervouet, Jean-Michel

    2016-07-01

    Bank erosion can be an important form of morphological adjustment in rivers. With the advances made in computational techniques, two-dimensional (2D) depth-averaged numerical models have become valuable tools for resolving many engineering problems dealing with sediment transport. The objective of this research work is to present a simple, new, bank-erosion operator that is integrated into a 2D Saint-Venant-Exner morphodynamic model. The numerical code is based on an unstructured grid of triangular elements and finite-element algorithms. The slope of each element in the grid is compared to the angle of repose of the bank material. Elements for which the slope is too steep are tilted to bring them to the angle of repose along a horizontal axis defined such that the volume loss above the axis is equal to the volume gain below, thus ensuring mass balance. The model performance is assessed using data from laboratory flume experiments and a scale model of the Old Rhine. For the flume experiment case with uniform bank material, relevant results are obtained for bank geometry changes. For the more challenging case (i.e. scale model of the Old Rhine with non-uniform bank material), the numerical model is capable of reproducing the main features of the bank failure, induced by the newly designed groynes, as well as the transport of the mobilized sediment material downstream. Some deviations between the computed results and measured data are, however, observed. They are ascribed to the effects of three-dimensional (3D) flow structures, pore pressure and cohesion, which are not considered in the present 2D model.

  1. GEL-STATE NMR OF BALL-MILLED WHOLE CELL WALLS IN DMSO-d6 USING 2D SOLUTION-STATE NMR SPECTROSCOPY

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant cell walls were used for obtaining 2D solution-state NMR spectra without actual solubilization or structural modification. Ball-milled whole cell walls were swelled directly in the NMR tube with DMSO-d6 where they formed a gel. There are relatively few gel-state NMR studies. Most have involved...

  2. Characterization of a novel NKG2D and NKp46 double-mutant mouse reveals subtle variations in the NK cell repertoire

    PubMed Central

    Sheppard, Sam; Triulzi, Chiara; Ardolino, Michele; Serna, Daniel; Zhang, Lily; Raulet, David H.

    2013-01-01

    The immunoreceptors NKG2D and NKp46 are known for their capacity to activate natural killer (NK) cell cytotoxicity and secretory responses in the contexts of tumors and infections, yet their roles in NK cell education remain unclear. Here, we provide the first characterization of mice deficient for both NKG2D and NKp46 receptors to address the relevance of their concomitant absence during NK cell development and function. Our findings reveal that NK cells develop normally in double-mutant (DKO) mice. Mice lacking NKG2D but not NKp46 showed subtle differences in the percentages of NK cells expressing inhibitory Ly49 receptors and the adhesion molecule DNAM-1. A slightly increased percentage of terminally differentiated NK cells and functional response to in vitro stimuli was observed in some experiments. These alterations were modest and did not affect NK cell function in vivo in response to mouse cytomegalovirus infection. NKp46 deficiency alone, or in combination with NKG2D deficiency, had no effect on frequency or function of NK cells. PMID:23649470

  3. Proteomics Approaches for Identification of Tumor Relevant Protein Targets in Pulmonary Squamous Cell Carcinoma by 2D-DIGE-MS

    PubMed Central

    Yiping, Guo; Yang, Song; Xiaoyu, Qi; Zhuzhu, Guan; Xiaohan, Yang; Xin, Zhou; Liyan, Xue; Shujuan, Shao

    2014-01-01

    Potential markers for progression of pulmonary squamous cell carcinoma (SCC) were identified by examining samples of lung SCC and adjacent normal tissues using a combination of fluorescence two-dimensional difference gel electrophoresis (2D-DIGE), matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and electrospray ionization quadrupole-time of flight mass spectrometry (ESI-Q-TOF). The PANTHER System was used for gel image based quantification and statistical analysis. An analysis of proteomic data revealed that 323 protein spots showed significantly different levels of expression (P≤0.05) in lung SCC tissue compared to expression in normal lung tissue. A further analysis of these protein spots by MALDI-TOF-MS identified 81 different proteins. A systems biology approach was used to map these proteins to major pathways involved in numerous cellular processes, including localization, transport, cellular component organization, apoptosis, and reproduction. Additionally, the expression of several proteins in lung SCC and normal tissues was examined using immunohistochemistry and western blot. The functions of individual proteins are being further investigated and validated, and the results might provide new insights into the mechanism of lung SCC progression, potentially leading to the design of novel diagnostic and therapeutic strategies. PMID:24740010

  4. Effect of random structure on permeability and heat transfer characteristics for flow in 2D porous medium based on MRT lattice Boltzmann method

    NASA Astrophysics Data System (ADS)

    Yang, PeiPei; Wen, Zhi; Dou, RuiFeng; Liu, Xunliang

    2016-08-01

    Flow and heat transfer through a 2D random porous medium are studied by using the lattice Boltzmann method (LBM). For the random porous medium, the influence of disordered cylinder arrangement on permeability and Nusselt number are investigated. Results indicate that the permeability and Nusselt number for different cylinder locations are unequal even with the same number and size of cylinders. New correlations for the permeability and coefficient b‧Den of the Forchheimer equation are proposed for random porous medium composed of Gaussian distributed circular cylinders. Furthermore, a general set of heat transfer correlations is proposed and compared with existing experimental data and empirical correlations. Our results show that the Nu number increases with the increase of the porosity, hence heat transfer is found to be accurate considering the effect of porosity.

  5. A methodology for linking 2D overland flow models with the sewer network model SWMM 5.1 based on dynamic link libraries.

    PubMed

    Leandro, Jorge; Martins, Ricardo

    2016-01-01

    Pluvial flooding in urban areas is characterized by a gradually varying inundation process caused by surcharge of the sewer manholes. Therefore urban flood models need to simulate the interaction between the sewer network and the overland flow in order to accurately predict the flood inundation extents. In this work we present a methodology for linking 2D overland flow models with the storm sewer model SWMM 5. SWMM 5 is a well-known free open-source code originally developed in 1971. The latest major release saw its structure re-written in C ++ allowing it to be compiled as a command line executable or through a series of calls made to function inside a dynamic link library (DLL). The methodology developed herein is written inside the same DLL in C + +, and is able to simulate the bi-directional interaction between both models during simulation. Validation is done in a real case study with an existing urban flood coupled model. The novelty herein is that the new methodology can be added to SWMM without the need for editing SWMM's original code. Furthermore, it is directly applicable to other coupled overland flow models aiming to use SWMM 5 as the sewer network model. PMID:27332848

  6. Downregulation of Ubiquitin-conjugating Enzyme UBE2D3 Promotes Telomere Maintenance and Radioresistance of Eca-109 Human Esophageal Carcinoma Cells

    PubMed Central

    Yang, Hui; Wu, Lin; Ke, Shaobo; Wang, Wenbo; Yang, Lei; Gao, Xiaojia; Fang, Hongyan; Yu, Haijun; Zhong, Yahua; Xie, Conghua; Zhou, Fuxiang; Zhou, Yunfeng

    2016-01-01

    Ubiquitin-conjugating enzyme UBE2D3 is an important member of the ubiquitin-proteasome pathways. Our previous study showed that the expression of UBE2D3 was negatively related to human telomerase reverse transcriptase (hTERT) and radioresistance in human breast cancer cells. However, in esophageal carcinoma, the exact effects and mechanisms of UBE2D3 in radioresistance remain unclear. This study shows that UBE2D3 knockdown was associated with significant increases in radioresistance to X-rays, telomerase activity, telomere length, and telomere shelterins. UBE2D3 knockdown-mediated radioresistance was related to a decrease in the spontaneous and ionizing radiation-induced apoptosis, resulting from a decrease in the Bax/Bcl-2 ratio. Furthermore, UBE2D3 downregulation was associated with increased G1-S phase transition and prolonged IR-induced G2/M arrest through over expression of cyclin D1, decrease of CDC25A expression and promotion of the ATM/ATR-Chk1-CDC25C pathway. Moreover, UBE2D3 downregulation reduced spontaneous DNA double-strand breaks and accelerated the repair of DNA damage induced by IR. The current data thus demonstrate that UBE2D3 downregulation enhances radioresistance by increased telomere homeostasis and prolonged IR-induced G2/M arrest, but decreases the IR-induced apoptosis and the number of DNA damage foci. These results suggest that UBE2D3 might be a potential molecular target to improve radiotherapy effects in esophageal carcinoma. PMID:27326259

  7. 3D rotating wall vessel and 2D cell culture of four veterinary virus pathogens: A comparison of virus yields, portions of infectious particles and virus growth curves.

    PubMed

    Malenovská, Hana

    2016-02-01

    Only very few comparative studies have been performed that evaluate general trends of virus growth under 3D in comparison with 2D cell culture conditions. The aim of this study was to investigate differences when four animal viruses are cultured in 2D and 3D. Suid herpesvirus 1 (SuHV-1), Vesicular stomatitis virus (VSIV), Bovine adenovirus (BAdV) and Bovine parainfluenza 3 virus (BPIV-3) were cultivated in 3D rotating wall vessels (RWVs) and conventional 2D cultures. The production of virus particles, the portion of infectious particles, and the infectious growth curves were compared. For all viruses, the production of virus particles (related to cell density), including the non-infectious ones, was lower in 3D than in 2D culture. The production of only infectious particles was significantly lower in BAdV and BPIV-3 in 3D cultures in relation to cell density. The two cultivation approaches resulted in significantly different virus particle-to-TCID50 ratios in three of the four viruses: lower in SuHV-1 and BPIV-3 and higher in BAdV in 3D culture. The infectious virus growth rates were not significantly different in all viruses. Although 3D RWV culture resulted in lower production of virus particles compared to 2D systems, the portion of infectious particles was higher for some viruses.

  8. UL16-binding proteins, novel MHC class I-related proteins, bind to NKG2D and activate multiple signaling pathways in primary NK cells.

    PubMed

    Sutherland, Claire L; Chalupny, N Jan; Schooley, Kenneth; VandenBos, Tim; Kubin, Marek; Cosman, David

    2002-01-15

    The UL16-binding proteins (ULBPs) are a novel family of MHC class I-related molecules that were identified as targets of the human CMV glycoprotein, UL16. We have previously shown that ULBP expression renders a relatively resistant target cell sensitive to NK cytotoxicity, presumably by engaging NKG2D, an activating receptor expressed by NK and other immune effector cells. In this study we show that NKG2D is the ULBP counterstructure on primary NK cells and that its expression is up-regulated by IL-15 stimulation. Soluble forms of ULBPs induce marked protein tyrosine phosphorylation, and activation of the Janus kinase 2, STAT5, extracellular signal-regulated kinase, mitogen-activated protein kinase, and phosphatidylinositol 3-kinase (PI 3-kinase)/Akt signal transduction pathways. ULBP-induced activation of Akt and extracellular signal-regulated kinase and ULBP-induced IFN-gamma production are blocked by inhibitors of PI 3-kinase, consistent with the known binding of PI 3-kinase to DAP10, the membrane-bound signal-transducing subunit of the NKG2D receptor. While all three ULBPs activate the same signaling pathways, ULBP3 was found to bind weakly and to induce the weakest signal. In summary, we have shown that NKG2D is the ULBP counterstructure on primary NK cells and for the first time have identified signaling pathways that are activated by NKG2D ligands. These results increase our understanding of the mechanisms by which NKG2D activates immune effector cells and may have implications for immune surveillance against pathogens and tumors. PMID:11777960

  9. BET_VH probabilistic assessment of pyroclastic flows hazard at El Misti volcano, South Peru, based on geological record and numerical simulations with TITAN2D

    NASA Astrophysics Data System (ADS)

    Constantinescu, R.; Thouret, J. C.; Sandri, L.; Irimus, I. A.; Stefanescu, R.

    2012-04-01

    Pyroclastic density currents, which include pyroclastic surges and pyroclastic flows (PFs), are among the most dangerous volcanic phenomena. We present a probabilistic hazard assessment of the PFs generated from eruptive column collapse at El Misti volcano (5822 m) in South Peru. The high relief of the cone, the location of the city of Arequipa (~1,000,000 people) on two large volcanoclastic fans and the H (3.5 km)/L (17 km) ratio (0.2) between the summit and the city center, make PFs a direct threat. We consider three eruption scenario sizes: small Vulcanian/Phreatomagmatic (VEI 2), medium Sub-Plinian (VEI 3-4), and large Plinian (VEI 4+). We use the Event-Tree approach in a Bayesian scheme with BET_VH (Bayesian Event Tree for Volcanic Hazard) software. Quantitative data that stem from numerical simulations from TITAN2D (termed prior models) and from stratigraphic record (termed past data) are input to BET_VH, which enables us to compute the probabilities (in a 1-year time window) of (i) having an eruption (ii) in a selected location/vent (iii) of a specific size, (iv) and that this eruption will produce PFs (v) that will reach a location of interest around El Misti. TITAN2D simulation runs, expressed as color-coded thicknesses of PDC deposits, fit well the extent of past PFs deposits, including thick confined deposits (0.5-7 m) in the Rio Chili canyon and its tributary ravines (Quebradas San Lazaro, Huarangal and Agua Salada).The unconfined, thinner (≤10cm) deposits, as displayed by simulation runs on the interfluves, is attributed to ash-cloud surges. Such thin, fine ash deposits have not been emphasized in geological maps either because they have been removed away or remain yet unrecognized. The simulated Vulcanian flows, restricted to the upper part of the cone, become confined (0.1-1m thick) in the ravines which converge towards each of the three Quebradas. The simulated Subplinian PF deposits reach 0.1 to 1 m thick in the Quebradas and 1-4 m WNW of El

  10. Lysis of HIV-1 Infected Autologous CD4+ Primary T cells by Interferon-alpha Activated NK cells Requires NKp46 and NKG2D

    PubMed Central

    Tomescu, Costin; Mavilio, Domenico; Montaner, Luis J.

    2015-01-01

    Objective Autologous HIV-1 infected CD4+ primary T cells (aHIV+CD4) have been shown to be largely resistant to Natural Killer (NK) cell mediated lysis due to viral strategies of immune evasion. We have previously shown that a pre-activation of NK cells with Plasmacytoid Dendritic Cells can significantly augment lysis of aHIV+CD4 through a mechanism dependent on Interferon-alpha (IFN-α). Design The goal of the present study is to identify the specific NK activating receptors involved in NK lysis of aHIV+CD4 following IFN-α activation. Methods PBMC were incubated with aHIV+CD4 to induce the secretion of endogenous levels of IFN-α and drive NK activation. We then utilized a standard chromium lysis assay to assess the degree of IFN-α activated lysis of aHIV+CD4 in the presence or absence of masking antibodies to a panel of NK activating receptors and co-receptors. Results Direct recognition of HIV-1 infected, but not uninfected, autologous CD4+ primary T cells by PBMC induced the secretion IFN-α (Median 2280 pg/ml, p<0.001, n=9) that, in turn, activated NK cells (p<0.001, n=12) and significantly increased their cytolytic potential against aHIV+CD4 (p<0.01, n=12). The masking of NKp46 (p<0.01, n=8) and NKG2D (p<0.05, n=8), but not 2B4, NTBA, NKp30 or NKp44, significantly reduced IFN-α activated lysis of aHIV+CD4. Conclusions Taken together, these results demonstrate that endogenous levels of IFN-α secreted by pDCs induce NK cells to lyse aHIV+CD4 via the engagement of NKp46 and NKG2D. PMID:26372382

  11. A Rapid and Efficient 2D/3D Nuclear Segmentation Method for Analysis of Early Mouse Embryo and Stem Cell Image Data

    PubMed Central

    Lou, Xinghua; Kang, Minjung; Xenopoulos, Panagiotis; Muñoz-Descalzo, Silvia; Hadjantonakis, Anna-Katerina

    2014-01-01

    Summary Segmentation is a fundamental problem that dominates the success of microscopic image analysis. In almost 25 years of cell detection software development, there is still no single piece of commercial software that works well in practice when applied to early mouse embryo or stem cell image data. To address this need, we developed MINS (modular interactive nuclear segmentation) as a MATLAB/C++-based segmentation tool tailored for counting cells and fluorescent intensity measurements of 2D and 3D image data. Our aim was to develop a tool that is accurate and efficient yet straightforward and user friendly. The MINS pipeline comprises three major cascaded modules: detection, segmentation, and cell position classification. An extensive evaluation of MINS on both 2D and 3D images, and comparison to related tools, reveals improvements in segmentation accuracy and usability. Thus, its accuracy and ease of use will allow MINS to be implemented for routine single-cell-level image analyses. PMID:24672759

  12. 25(OH)D Is Effective to Repress Human Cholangiocarcinoma Cell Growth through the Conversion of 25(OH)D to 1α,25(OH)2D3

    PubMed Central

    Chiang, Kun-Chun; Yeh, Chun-Nan; Huang, Cheng-Cheng; Yeh, Ta-Sen; S. Pang, Jong-Hwei; Hsu, Jun-Te; Chen, Li-Wei; Kuo, Sheng-Fong; Kittaka, Atsushi; Chen, Tai C.; Juang, Horng-Heng

    2016-01-01

    Cholangiocarcinoma (CCA) is a devastating disease without effective treatments. 1α,25(OH)2D3, the active form of Vitamin D, has emerged as a new anti-cancer regimen. However, the side effect of hypercalcemia impedes its systemic administration. 25(OH)D is biologically inert and needs hydroxylation by CYP27B1 to form 1α,25(OH)2D3, which is originally believed to only take place in kidneys. Recently, the extra-renal expression of CYP27B1 has been identified and in vitro conversion of 25(OH)D to 1α,25(OH)2D3 has been found in some cancer cells with CYP27B1 expression. In this study, CYP27B1 expression was demonstrated in CCA cells and human CCA specimens. 25(OH)D effectively represses SNU308 cells growth, which was strengthened or attenuated as CYP27B1 overexpression or knockdown. Lipocalcin-2 (LCN2) was also found to be repressed by 25(OH)D. After treatment with 800 ng/mL 25(OH)D, the intracellular 1α,25(OH)2D3 concentration was higher in SNU308 cells with CYP27B1 overexpression than wild type SNU308 cells. In a xenograft animal experiment, 25(OH)D, at a dose of 6 μg/kg or 20 μg/kg, significantly inhibited SNU308 cells’ growth without inducing obvious side effects. Collectively, our results indicated that SNU308 cells were able to convert 25(OH)D to 1α,25(OH)2D3 and 25(OH)D CYP27B1 gene therapy could be deemed as a promising therapeutic direction for CCA. PMID:27529229

  13. Ubiquitin-dependent endocytosis of NKG2D-DAP10 receptor complexes activates signaling and functions in human NK cells.

    PubMed

    Quatrini, Linda; Molfetta, Rosa; Zitti, Beatrice; Peruzzi, Giovanna; Fionda, Cinzia; Capuano, Cristina; Galandrini, Ricciarda; Cippitelli, Marco; Santoni, Angela; Paolini, Rossella

    2015-10-27

    Cytotoxic lymphocytes share the presence of the activating receptor NK receptor group 2, member D (NKG2D) and the signaling-competent adaptor DNAX-activating protein 10 (DAP10), which together play an important role in antitumor immune surveillance. Ligand stimulation induces the internalization of NKG2D-DAP10 complexes and their delivery to lysosomes for degradation. In experiments with human NK cells and cell lines, we found that the ligand-induced endocytosis of NKG2D-DAP10 depended on the ubiquitylation of DAP10, which was also required for degradation of the internalized complexes. Moreover, through combined biochemical and microscopic analyses, we showed that ubiquitin-dependent receptor endocytosis was required for the activation of extracellular signal-regulated kinase (ERK) and NK cell functions, such as the secretion of cytotoxic granules and the inflammatory cytokine interferon-γ. These results suggest that NKG2D-DAP10 endocytosis represents a means to decrease cell surface receptor abundance, as well as to control signaling outcome in cytotoxic lymphocytes.

  14. Differential effects of MAPK pathway inhibitors on migration and invasiveness of BRAF(V600E) mutant thyroid cancer cells in 2D and 3D culture.

    PubMed

    Ingeson-Carlsson, Camilla; Martinez-Monleon, Angela; Nilsson, Mikael

    2015-11-01

    Tumor microenvironment influences targeted drug therapy. In this study we compared drug responses to RAF and MEK inhibitors on tumor cell migration in 2D and 3D culture of BRAF(V600E) mutant cell lines derived from human papillary (BCPAP) and anaplastic (SW1736) thyroid carcinomas. Scratch wounding was compared to a double-layered collagen gel model developed for analysis of directed tumor cell invasion during prolonged culture. In BCPAP both PLX4720 and U0126 inhibited growth and migration in 2D and decreased tumor cell survival in 3D. In SW1736 drugs had no effect on migration in 2D but decreased invasion in 3D, however this related to reduced growth. Dual inhibition of BRAF(V600E) and MEK reduced but did not prevent SW1736 invasion although rebound phosphorylation of ERK in response to PLX4720 was blocked by U0126. These findings indicate that anti-tumor drug effects in vitro differ depending on culture conditions (2D vs. 3D) and that the invasive features of anaplastic thyroid cancer depend on non-MEK mechanism(s).

  15. SUTRA: A model for 2D or 3D saturated-unsaturated, variable-density ground-water flow with solute or energy transport

    USGS Publications Warehouse

    Voss, Clifford I.; Provost, A.M.

    2002-01-01

    SUTRA (Saturated-Unsaturated Transport) is a computer program that simulates fluid movement and the transport of either energy or dissolved substances in a subsurface environment. This upgraded version of SUTRA adds the capability for three-dimensional simulation to the former code (Voss, 1984), which allowed only two-dimensional simulation. The code employs a two- or three-dimensional finite-element and finite-difference method to approximate the governing equations that describe the two interdependent processes that are simulated: 1) fluid density-dependent saturated or unsaturated ground-water flow; and 2) either (a) transport of a solute in the ground water, in which the solute may be subject to: equilibrium adsorption on the porous matrix, and both first-order and zero-order production or decay; or (b) transport of thermal energy in the ground water and solid matrix of the aquifer. SUTRA may also be used to simulate simpler subsets of the above processes. A flow-direction-dependent dispersion process for anisotropic media is also provided by the code and is introduced in this report. As the primary calculated result, SUTRA provides fluid pressures and either solute concentrations or temperatures, as they vary with time, everywhere in the simulated subsurface system. SUTRA flow simulation may be employed for two-dimensional (2D) areal, cross sectional and three-dimensional (3D) modeling of saturated ground-water flow systems, and for cross sectional and 3D modeling of unsaturated zone flow. Solute-transport simulation using SUTRA may be employed to model natural or man-induced chemical-species transport including processes of solute sorption, production, and decay. For example, it may be applied to analyze ground-water contaminant transport problems and aquifer restoration designs. In addition, solute-transport simulation with SUTRA may be used for modeling of variable-density leachate movement, and for cross sectional modeling of saltwater intrusion in

  16. Contrasting Effects of the Cytotoxic Anticancer Drug Gemcitabine and the EGFR Tyrosine Kinase Inhibitor Gefitinib on NK Cell-Mediated Cytotoxicity via Regulation of NKG2D Ligand in Non-Small-Cell Lung Cancer Cells

    PubMed Central

    Okita, Riki; Wolf, Diana; Yasuda, Koichiro; Maeda, Ai; Yukawa, Takuro; Saisho, Shinsuke; Shimizu, Katsuhiko; Yamaguchi, Yoshiyuki; Oka, Mikio; Nakayama, Eiichi; Lundqvist, Andreas; Kiessling, Rolf; Seliger, Barbara; Nakata, Masao

    2015-01-01

    Introduction Several cytotoxic anticancer drugs inhibit DNA replication and/or mitosis, while EGFR tyrosine kinase inhibitors inactivate EGFR signalling in cancer cell. Both types of anticancer drugs improve the overall survival of the patients with non-small-cell lung cancer (NSCLC), although tumors often become refractory to this treatment. Despite several mechanisms by which the tumors become resistant having been described the effect of these compounds on anti-tumor immunity remains largely unknown. Methods This study examines the effect of the cytotoxic drug Gemcitabine and the EGFR tyrosine kinase inhibitor Gefitinib on the expression of NK group 2 member D (NKG2D) ligands as well as the sensitivity of NSCLC cells to the NK-mediated lysis. Results We demonstrate that Gemcitabine treatment leads to an enhanced expression, while Gefitinib downregulated the expression of molecules that act as key ligands for the activating receptor NKG2D and promote NK cell-mediated recognition and cytolysis. Gemcitabine activated ATM and ATM- and Rad-3-related protein kinase (ATR) pathways. The Gemcitabine-induced phosphorylation of ATM as well as the upregulation of the NKG2D ligand expression could be blocked by an ATM-ATR inhibitor. In contrast, Gefitinib attenuated NKG2D ligand expression. Silencing EGFR using siRNA or addition of the PI3K inhibitor resulted in downregulation of NKG2D ligands. The observations suggest that the EGFR/PI3K pathway also regulates the expression of NKG2D ligands. Additionally, we showed that both ATM-ATR and EGFR regulate MICA/B via miR20a. Conclusion In keeping with the effect on NKG2D expression, Gemcitabine enhanced NK cell-mediated cytotoxicity while Gefitinib attenuated NK cell killing in NSCLC cells. PMID:26439264

  17. miR-103 promotes 3T3-L1 cell adipogenesis through AKT/mTOR signal pathway with its target being MEF2D.

    PubMed

    Li, Meihang; Liu, Zhenjiang; Zhang, Zhenzhen; Liu, Guannv; Sun, Shiduo; Sun, Chao

    2015-03-01

    MicroRNAs are small non-coding RNAs that partially bind to the 3' untranslated (3'UTR) regions of target genes in animals and regulate protein production of the target transcripts. MiR-103 has been confirmed to play a critical role in lipid metabolism, however, the target genes and signaling pathway regulated by miR-103 is still unclear. In our experiment, we observed a positive function of miR-103 on the adipogenic differentiation of 3T3-L1 pre-adipocyte. Furthermore, we proved that this function of miR-103 worked through activating AKT/mTOR signal pathway and impairing target gene MEF2D. By inhibiting and over-expressing the MEF2D gene, we found that MEF2D had a negative role in regulating adipocyte key genes, and this function of MEF2D could be impaired by miR-103. In conclusion, we found that miR-103 can promote 3T3-L1 cells differentiation by targeting MEF2D and activating AKT/mTOR signal pathway. These results will shed a light on further study of microRNAs. PMID:25400071

  18. Effects of 1α,25-(OH)2D3 on the formation and activity of osteoclasts in RAW264.7 cells.

    PubMed

    Gu, Jianhong; Tong, Xi-Shuai; Chen, Guo-Hong; Wang, Dong; Chen, Yang; Yuan, Yan; Liu, Xue-Zhong; Bian, Jian-Chun; Liu, Zong-Ping

    2015-08-01

    The hormonally active form of vitamin D3, 1α,25-(OH)2D3, has an important role in bone metabolism. This study examined the effects of 1α,25-(OH)2D3 on the ability of two cytokines, receptor activator of nuclear factor-κB ligand (RANKL) and macrophage-colony stimulating factor (M-CSF), to induce RAW 264.7 cells to form osteoclasts. A TRAP histochemical staining assay and bone resorption analysis were used to identify the rate of formation and activity of osteoclasts. The numbers of osteoclasts formed, and their bone resorption activity, was enhanced by the addition of 1α,25-(OH)2D3. The expression levels of osteoclast-specific proteins that are essential for bone resorption, integrin β3, V-ATPase, CAII, CTSK, TRAP and MMP-9, were detected by western blotting. During 48 h, the expression levels of all these proteins significantly increased. Quantitative real-time polymerase chain reaction was used to determine the expression levels of the transcription factors, c-Fos and NFATcl. The expression levels of c-Fos and NFATc1 also increased 24h after treatment with 1α,25-(OH)2D3. These results suggest that 1α,25-(OH)2D3 can regulate bone metabolism by directly enhancing the formation and maturation of osteoclasts.

  19. Design-support and performance estimation using HYDRUS/CW2D: a horizontal flow constructed wetland for polishing SBR effluent.

    PubMed

    Pálfy, Tamás Gábor; Gribovszki, Zoltán; Langergraber, Günter

    2015-01-01

    The 4,000 PE (700 m(3)/d) wastewater treatment plant at Balf, Hungary was based on sequencing batch reactor technology with phosphorus precipitation as the tertiary step. Its effluent met quality thresholds on average, with above-threshold peaks mainly in winter. The HYDRUS/CW2D model package (PC Progress s.r.o.) was used to simulate the treatment performance of a horizontal flow constructed wetland for polishing effluent. The goal of this study was to provide design-supportive information about the suitability of the proposed wetland and to prove the applicability of the computational tool used to gain that information. The simulations showed that the wetland with the proposed layout could not tackle peaks in NH4-N. Other effluent thresholds could be met, including chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total nitrogen, total inorganic nitrogen and total phosphorus. The tool was unable to simulate COD and BOD5 removal in periods when the electron acceptors were depleted because anaerobic processes are not modelled. Using a tool of such complexity for designing carries excessive work demands and involves many uncertainties. The simulation study highlighted that the model used could still facilitate the design of an effective system by showing the weaknesses of a test scenario as it was demonstrated. PMID:25860697

  20. Design-support and performance estimation using HYDRUS/CW2D: a horizontal flow constructed wetland for polishing SBR effluent.

    PubMed

    Pálfy, Tamás Gábor; Gribovszki, Zoltán; Langergraber, Günter

    2015-01-01

    The 4,000 PE (700 m(3)/d) wastewater treatment plant at Balf, Hungary was based on sequencing batch reactor technology with phosphorus precipitation as the tertiary step. Its effluent met quality thresholds on average, with above-threshold peaks mainly in winter. The HYDRUS/CW2D model package (PC Progress s.r.o.) was used to simulate the treatment performance of a horizontal flow constructed wetland for polishing effluent. The goal of this study was to provide design-supportive information about the suitability of the proposed wetland and to prove the applicability of the computational tool used to gain that information. The simulations showed that the wetland with the proposed layout could not tackle peaks in NH4-N. Other effluent thresholds could be met, including chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total nitrogen, total inorganic nitrogen and total phosphorus. The tool was unable to simulate COD and BOD5 removal in periods when the electron acceptors were depleted because anaerobic processes are not modelled. Using a tool of such complexity for designing carries excessive work demands and involves many uncertainties. The simulation study highlighted that the model used could still facilitate the design of an effective system by showing the weaknesses of a test scenario as it was demonstrated.

  1. Increased sMICA and TGFβ1 levels in HNSCC patients impair NKG2D-dependent functionality of activated NK cells

    PubMed Central

    Klöß, Stephan; Chambron, Nicole; Gardlowski, Tanja; Arseniev, Lubomir; Koch, Joachim; Esser, Ruth; Glienke, Wolfgang; Seitz, Oliver; Köhl, Ulrike

    2015-01-01

    Disseminated head-and-neck squamous cell carcinoma (HNSCC) escapes immune surveillance and thus frequently manifests as fatal disease. Here, we report on the distribution of distinct immune cell subpopulations, natural killer (NK) cell cytotoxicity and tumor immune escape mechanisms (TIEMs) in 55 HNSCC patients, either at initial diagnosis or present with tumor relapse. Compared to healthy controls, the regulatory NK cells and the ratio of pro/anti-inflammatory cytokines were decreased in HNSCC patients, while soluble major histocompatibility complex Class I chain-related peptide A (sMICA) and transforming growth factor β1 (TGFβ1) plasma levels were markedly elevated. Increased sMICA and TGFβ1 concentrations correlated with tumor progression and staging characteristics in 7 follow-up HNSCC patients, with significantly elevated levels of both soluble factors from the time of initial diagnosis to that of relapse. Patient plasma containing elevated sMICA and TGFβ1 markedly impaired NKG2D-dependent cytotoxicity against HNSCC cells upon incubation with patient-derived and IL-2 activated NK cells vs. those derived from healthy donors. Decreased antitumor recognition was accompanied by reduced NKG2D expression on the NK cell surface and an enhanced caspase-3 activity. In-vitro blocking and neutralization experiments demonstrated a synergistic negative impact of sMICA and TGFβ1 on NK cell functionality. Although we previously showed the feasibility and safety of transfer of allogeneic donor NK cells in a prior clinical study encompassing various leukemia and tumor patients, our present results suggest the need for caution regarding the sole use of adoptive NK cell transfer. The presence of soluble NKG2D ligands in the plasma of HNSCC patients and the decreased NK cell cytotoxicity due to several factors, especially TGFβ1, indicates timely depletion of these immunosuppressing molecules may promote NK cell-based immunotherapy. PMID:26451327

  2. DNT cell inhibits the growth of pancreatic carcinoma via abnormal expressions of NKG2D and MICA in vivo.

    PubMed

    Xu, Hong; Zhu, Xing-Xing; Chen, Jiong

    2016-01-01

    This research aimed to investigate the effects of natural killer group 2 member D (NKG2D) and its ligands major histocompatibility complex class I chain-related molecules A(MICA) in DNT cell killing pancreatic carcinoma. Antibodies adsorption was used to separate DNT cell from human peripheral blood. Human pancreatic tumor models were established via implanting BXPC-3 cells into nude mice. Then randomly divided mice into blank group, gemcitabine group and DNT group. Mice weights and mice tumor volumes were measured every 5 days. 50 days later mice were euthanized at cervical dislocation method. Tumor weights were measured. Relative tumor volume and tumor inhibition rate were calculated. Western blot and qPCR were used to detect the expressions of NKG2D and MICA in the transplanted tumors of the three groups. DNT cell significantly increased over time. The blank group tumor volume and weight were significantly larger than the other groups (p < 0.001, p < 0.001), but there were no significantly difference between DNT group and gemcitabine group (p > 0.05). Gemcitabine and DNT cell tumor inhibition rate were 40.4% and 35.5%. Western blot and qPCR showed that MICA mRNA and protein levels in blank group were significantly higher than DNT group (p = 0.001, p = 0.003). NKG2D mRNA and protein levels in blank group were significantly lower than DNT cells group (p < 0.001, p = 0.001). In conclusion DNT cell can significantly inhibit the growth of pancreatic carcinoma in vivo, and the mechanism may be involved in abnormal expressions of MICA and NKG2D. PMID:26616050

  3. DNT cell inhibits the growth of pancreatic carcinoma via abnormal expressions of NKG2D and MICA in vivo.

    PubMed

    Xu, Hong; Zhu, Xing-Xing; Chen, Jiong

    2016-01-01

    This research aimed to investigate the effects of natural killer group 2 member D (NKG2D) and its ligands major histocompatibility complex class I chain-related molecules A(MICA) in DNT cell killing pancreatic carcinoma. Antibodies adsorption was used to separate DNT cell from human peripheral blood. Human pancreatic tumor models were established via implanting BXPC-3 cells into nude mice. Then randomly divided mice into blank group, gemcitabine group and DNT group. Mice weights and mice tumor volumes were measured every 5 days. 50 days later mice were euthanized at cervical dislocation method. Tumor weights were measured. Relative tumor volume and tumor inhibition rate were calculated. Western blot and qPCR were used to detect the expressions of NKG2D and MICA in the transplanted tumors of the three groups. DNT cell significantly increased over time. The blank group tumor volume and weight were significantly larger than the other groups (p < 0.001, p < 0.001), but there were no significantly difference between DNT group and gemcitabine group (p > 0.05). Gemcitabine and DNT cell tumor inhibition rate were 40.4% and 35.5%. Western blot and qPCR showed that MICA mRNA and protein levels in blank group were significantly higher than DNT group (p = 0.001, p = 0.003). NKG2D mRNA and protein levels in blank group were significantly lower than DNT cells group (p < 0.001, p = 0.001). In conclusion DNT cell can significantly inhibit the growth of pancreatic carcinoma in vivo, and the mechanism may be involved in abnormal expressions of MICA and NKG2D.

  4. NKG2D performs two functions in invariant NKT cells: Direct TCR-independent activation of NK-like cytolysis, and co-stimulation of activation by CD1d

    PubMed Central

    Kuylenstierna, Carlotta; Björkström, Niklas K.; Andersson, Sofia K.; Sahlström, Peter; Bosnjak, Lidija; Paquin-Proulx, Dominic; Malmberg, Karl-Johan; Ljunggren, Hans-Gustaf; Moll, Markus; Sandberg, Johan K.

    2012-01-01

    Invariant NKT cells are important in the activation and regulation of immune responses. They can also function as CD1d-restricted killer cells. However, the role of activating innate NK cell receptors expressed on NKT cells in triggering cytolytic function is poorly characterized. Here, we initially confirmed that the cellular stress-ligand receptor NKG2D is expressed on CD4− NKT cells, whereas most CD4+ NKT cells lack this receptor. Interestingly, NKG2D+ NKT cells frequently expressed perforin, and both NKG2D and perforin localized at the site of contact with NKG2D ligand-expressing target cells. CD4− NKT cells degranulated in response to NKG2D engagement in a redirected activation assay independently of stimulation via their invariant TCR. NKT cells killed P815 cells coated with anti-NKG2D mAb and CD1d-negative K562 tumor target cells in an NKG2D-dependent manner. Furthermore, NKG2D engagement co-stimulated TCR-mediated NKT cell activation in response to endogenous CD1d-presented ligands or suboptimal levels of anti-CD3 triggering. These data indicate that the CD4− subset of human NKT cells can mediate direct lysis of target cells via NKG2D engagement independently of CD1d, and that NKG2D also functions as a co-stimulatory receptor in these cells. NKG2D thus plays both a direct and a co-stimulatory role in the activation of NKT cells. PMID:21590763

  5. Tamoxifen and raloxifene modulate gap junction coupling during early phases of retinoic acid-dependent neuronal differentiation of NTera2/D1 cells

    PubMed Central

    Dahm, Liane; Klugmann, Fanny; Gonzalez-Algaba, Angeles

    2010-01-01

    Gap junctions (GJ) represent a cellular communication system known to influence neuronal differentiation and survival. To assess a putative role of this system for neural effects of tamoxifen (TAM) and raloxifene (RAL), we used the human teratocarcinoma cell line NTera2/D1, retinoic acid (RA)-dependent neuronal differentiation of which is regulated by gap junctions formed of connexin43 (Cx43). As demonstrated by Western blot analysis, concentrations above 1 µmol/l for TAM, and 0.1 µmol/l for RAL lead to a temporary time- and concentration-dependent increase in Cx43 immunoreactivity, which reached a peak for TAM after 1 day and for RAL after 2 days. Immunocytochemical stainings revealed the increase in Cx43 immunoreactivity to result from an accumulation in intracellular compartments such as the Golgi apparatus or lysosomes. In addition, TAM and RAL were able to prevent the RA-dependent decrease of Cx43 immunoreactivity in NTera2/D1 cells, normally observed during neuronal differentiation. This suggested a suppression of neuronal differentiation to result from these substances. According to this, treatment of NTera2/D1 cells with 10 µmol/l TAM or RAL during weeks 1 and 2 of a 6 weeks RA-driven differentiation schedule impaired, whereas treatment during weeks 5 and 6 did not impair, neuronal differentiation of these cells. Modulation of GJ coupling between NTera2/D1 cells by TAM and RAL seems therefore to perturb early neuronal differentiation, whereas differentiated neurons in the mature brain seem to be not affected. These effects could be of importance for actions of TAM and RAL on early embryonic steps of nervous system formation. PMID:20437090

  6. Genotoxic Effects of Low- and High-LET Radiation on Human Epithelial Cells Grown in 2-D Versus 3-D Culture

    NASA Technical Reports Server (NTRS)

    Patel, Z. S.; Cucinotta, F. A.; Huff, J. L.

    2011-01-01

    Risk estimation for radiation-induced cancer relies heavily on human epidemiology data obtained from terrestrial irradiation incidents from sources such as medical and occupational exposures as well as from the atomic bomb survivors. No such data exists for exposures to the types and doses of high-LET radiation that will be encountered during space travel; therefore, risk assessment for space radiation requires the use of data derived from cell culture and animal models. The use of experimental models that most accurately replicate the response of human tissues is critical for precision in risk projections. This work compares the genotoxic effects of radiation on normal human epithelial cells grown in standard 2-D monolayer culture compared to 3-D organotypic co-culture conditions. These 3-D organotypic models mimic the morphological features, differentiation markers, and growth characteristics of fully-differentiated normal human tissue and are reproducible using defined components. Cultures were irradiated with 2 Gy low-LET gamma rays or varying doses of high-LET particle radiation and genotoxic damage was measured using a modified cytokinesis block micronucleus assay. Our results revealed a 2-fold increase in residual damage in 2 Gy gamma irradiated cells grown under organotypic culture conditions compared to monolayer culture. Irradiation with high-LET particle radiation gave similar results, while background levels of damage were comparable under both scenarios. These observations may be related to the phenomenon of "multicellular resistance" where cancer cells grown as 3-D spheroids or in vivo exhibit an increased resistance to killing by chemotherapeutic agents compared to the same cells grown in 2-D culture. A variety of factors are likely involved in mediating this process, including increased cell-cell communication, microenvironment influences, and changes in cell cycle kinetics that may promote survival of damaged cells in 3-D culture that would

  7. Proteomic analysis of doxorubicin-induced changes in the proteome of HepG2cells combining 2-D DIGE and LC-MS/MS approaches.

    PubMed

    Hammer, Elke; Bien, Sandra; Salazar, Manuela Gesell; Steil, Leif; Scharf, Christian; Hildebrandt, Petra; Schroeder, Henry W S; Kroemer, Heyo K; Völker, Uwe; Ritter, Christoph A

    2010-01-01

    HepG-2 cells are widely used as a cell model to investigate hepatocellular carcinomas and the effect of anticancer drugs such as doxorubicin, an effective antineoplastic agent, which has broad antitumoral activity against many solid and hematological malignancies. To investigate the effect of doxorubicin on the protein pattern, we used complementary proteomic workflows including 2-D gel-based and gel-free methods. The analysis of crude HepG2 cell extracts by 2-D DIGE provided data on 1835 protein spots which was then complemented by MS-centered analysis of stable isotope labeling by amino acids in cell culture-labeled cells. The monitoring of more than 1300 distinct proteins, including proteins of the membrane fraction provides the most comprehensive overview on the proteome of the widely used model cell line HepG2. Of the proteins monitored in total, 155 displayed doxorubicin-induced changes in abundance. Functional analysis revealed major influences of doxorubicin on proteins involved in protein synthesis, DNA damage control, electron transport/mitochondrial function, and tumor growth. The strongest decrease in level was found for proteins involved in DNA replication and protein synthesis, whereas proteins with a function in DNA damage control and oxidative stress management displayed increased levels following treatment with doxorubicin compared with control cells. Furthermore, the doxorubicin-associated increase in levels of multiple forms of keratins 8, 18, and 19 and other structural proteins revealed an influence on the cytoskeleton network.

  8. Glycosylated VCAM-1 isoforms revealed in 2D western blots of HUVECs treated with tumoral soluble factors of breast cancer cells

    PubMed Central

    2009-01-01

    Background Several common aspects of endothelial phenotype, such as the expression of cell adhesion molecules, are shared between metastasis and inflammation. Here, we analyzed VCAM-1 variants as biological markers of these two types of endothelial cell activation. With the combination of 2-DE and western blot techniques and the aid of tunicamycin, we analyzed N-glycosylation variants of VCAM-1 in primary human endothelial cells stimulated with either TNF or tumoral soluble factors (TSF's) derived from the human breast cancer cell line ZR75.30. Results Treatments induced a pro-adhesive endothelial phenotype. 2D western blots analysis of cells subjected to both treatments revealed the expression of the two known VCAM-1 isoforms and of previously unknown isoforms. In particular TSFZR75.30 induced an isoform with a relative molecular mass (Mr) and isoelectric point (pI) of 75-77 kDa and 5.0, respectively. Conclusion The unknown isoforms of VCAM-1 that were found to be overexpressed after treatment with TSF's compared with TNF, could serve as biomarkers to discriminate between inflammation and metastasis. 2D western blots revealed three new VCAM-1 isoforms expressed in primary human endothelial cells in response to TSF stimulation. Each of these isoforms varies in Mr and pI and could be the result of differential glycosylation states. PMID:19930605

  9. A robust cell counting approach based on a normalized 2D cross-correlation scheme for in-line holographic images.

    PubMed

    Ra, Ho-Kyeong; Kim, Hyungseok; Yoon, Hee Jung; Son, Sang Hyuk; Park, Taejoon; Moon, Sangjun

    2013-09-01

    To achieve the important aims of identifying and marking disease progression, cell counting is crucial for various biological and medical procedures, especially in a Point-Of-Care (POC) setting. In contrast to the conventional manual method of counting cells, a software-based approach provides improved reliability, faster speeds, and greater ease of use. We present a novel software-based approach to count in-line holographic cell images using the calculation of a normalized 2D cross-correlation. This enables fast, computationally-efficient pattern matching between a set of cell library images and the test image. Our evaluation results show that the proposed system is capable of quickly counting cells whilst reliably and accurately following human counting capability. Our novel approach is 5760 times faster than manual counting and provides at least 68% improved accuracy compared to other image processing algorithms. PMID:23839256

  10. 2D and 3D Stem Cell Models of Primate Cortical Development Identify Species-Specific Differences in Progenitor Behavior Contributing to Brain Size

    PubMed Central

    Otani, Tomoki; Marchetto, Maria C.; Gage, Fred H.; Simons, Benjamin D.; Livesey, Frederick J.

    2016-01-01

    Summary Variation in cerebral cortex size and complexity is thought to contribute to differences in cognitive ability between humans and other animals. Here we compare cortical progenitor cell output in humans and three nonhuman primates using directed differentiation of pluripotent stem cells (PSCs) in adherent two-dimensional (2D) and organoid three-dimensional (3D) culture systems. Clonal lineage analysis showed that primate cortical progenitors proliferate for a protracted period of time, during which they generate early-born neurons, in contrast to rodents, where this expansion phase largely ceases before neurogenesis begins. The extent of this additional cortical progenitor expansion differs among primates, leading to differences in the number of neurons generated by each progenitor cell. We found that this mechanism for controlling cortical size is regulated cell autonomously in culture, suggesting that primate cerebral cortex size is regulated at least in part at the level of individual cortical progenitor cell clonal output. PMID:27049876

  11. In Vitro Differentiation of Pluripotent Stem Cells into Functional β Islets Under 2D and 3D Culture Conditions and In Vivo Preclinical Validation of 3D Islets.

    PubMed

    Bose, Bipasha; Sudheer, P Shenoy

    2016-01-01

    Since the advent of pluripotent stem cells, (embryonic and induced pluripotent stem cells), applications of such pluripotent stem cells are of prime importance. Indeed, scientists are involved in studying the basic biology of pluripotent stem cells, but equal impetus is there to direct the pluripotent stem cells into multiple lineages for cell therapy applications. Scientists across the globe have been successful, to a certain extent, in obtaining cells of definitive endoderm and also pancreatic β islets by differentiating human pluripotent stem cells. Pluripotent stem cell differentiation protocols aim at mimicking in vivo embryonic development. As in vivo embryonic development is a complex process and involves interplay of multiple cytokines, the differentiation protocols also involve a stepwise use of multiple cytokines. Indeed the novel markers for pancreas organogenesis serve as the roadmaps to develop new protocols for pancreatic differentiation from pluripotent stem cells. Earliest developed protocols for pancreas differentiation involved "Nestin selection pathway," a pathway common for both neuronal and pancreatic differentiation lead to the generation of cells that were a combination of cells from neuronal lineage. Eventually with the discovery of hierarchy of β cell transcription factors like Pdx1, Pax4, and Nkx2.2, forced expression of such transcription factors proved successful in converting a pluripotent stem cell into a β cell. Protocols developed almost half a decade ago to the recent ones rather involve stepwise differentiations involving various cytokines and could generate as high as 25 % functional insulin-positive cells in vitro. Most advanced protocols for β islet differentiations from human pluripotent stem cells focused on 3D culture conditions, which reportedly produced 60-65 % functional β islet cells. Here, we describe the protocol for differentiation of human pluripotent stem cells into functional β cells under both 2D and 3D

  12. Marked change in microRNA expression during neuronal differentiation of human teratocarcinoma NTera2D1 and mouse embryonal carcinoma P19 cells

    SciTech Connect

    Hohjoh, Hirohiko Fukushima, Tatsunobu

    2007-10-19

    MicroRNAs (miRNAs) are small noncoding RNAs, with a length of 19-23 nucleotides, which appear to be involved in the regulation of gene expression by inhibiting the translation of messenger RNAs carrying partially or nearly complementary sequences to the miRNAs in their 3' untranslated regions. Expression analysis of miRNAs is necessary to understand their complex role in the regulation of gene expression during the development, differentiation and proliferation of cells. Here we report on the expression profile analysis of miRNAs in human teratocarcinoma NTere2D1, mouse embryonic carcinoma P19, mouse neuroblastoma Neuro2a and rat pheochromocytoma PC12D cells, which can be induced into differentiated cells with long neuritic processes, i.e., after cell differentiation, such that the resultant cells look similar to neuronal cells. The data presented here indicate marked changes in the expression of miRNAs, as well as genes related to neuronal development, occurred in the differentiation of NTera2D1 and P19 cells. Significant changes in miRNA expression were not observed in Neuro2a and PC12D cells, although they showed apparent morphologic change between undifferentiated and differentiated cells. Of the miRNAs investigated, the expression of miRNAs belonging to the miR-302 cluster, which is known to be specifically expressed in embryonic stem cells, and of miR-124a specific to the brain, appeared to be markedly changed. The miR-302 cluster was potently expressed in undifferentiated NTera2D1 and P19 cells, but hardly in differentiated cells, such that miR-124a showed an opposite expression pattern to the miR-302 cluster. Based on these observations, it is suggested that the miR-302 cluster and miR-124a may be useful molecular indicators in the assessment of degree of undifferentiation and/or differentiation in the course of neuronal differentiation.

  13. Mirrored serpentine flow channels for fuel cell

    SciTech Connect

    Rock, Jeffrey Allan

    2000-08-08

    A PEM fuel cell having serpentine flow field channels wherein the input/inlet legs of each channel border the input/inlet legs of the next adjacent channels in the same flow field, and the output/exit legs of each channel border the output/exit legs of the next adjacent channels in the same flow field. The serpentine fuel flow channels may be longer, and may contain more medial legs, than the serpentine oxidant flow channels.

  14. 2-D numerical simulations of groundwater flow, heat transfer and 4He transport — implications for the He terrestrial budget and the mantle helium heat imbalance

    NASA Astrophysics Data System (ADS)

    Castro, Maria Clara; Patriarche, Delphine; Goblet, Patrick

    2005-09-01

    Because helium and heat production results from a common source, a continental 4He crustal flux of 4.65 * 10 - 14 mol m - 2 s - 1 has been estimated based on heat flow considerations. In addition, because the observed mantle He / heat flux ratio at the proximity of mid-ocean ridges (6.6 * 10 - 14 mol J - 1 ) is significantly lower than the radiogenic production ratio (1.5 * 10 - 12 mol J - 1 ), the presence of a terrestrial helium-heat imbalance was suggested. The latter could be explained by the presence of a layered mantle in which removal of He is impeded from the lower mantle [R.K. O'Nions, E.R. Oxburgh, Heat and helium in the Earth, Nature 306 (1983) 429-431; E.R. Oxburgh, R.K. O'Nions, Helium loss, tectonics, and the terrestrial heat budget, Science 237 (1987) 1583-1588]. van Keken et al. [P.E. van Keken, C.J. Ballentine, D. Porcelli, A dynamical investigation of the heat and helium imbalance, Earth Planet, Sci. Lett. 188 (2001) 421-434] have recently claimed that the helium-heat imbalance remains a robust observation. Such conclusions, however, were reached under the assumption that a steady-state regime was in place for both tracers and that their transport properties are similar at least in the upper portion of the crust. Here, through 2-D simulations of groundwater flow, heat transfer and 4He transport carried out simultaneously in the Carrizo aquifer and surrounding formations in southwest Texas, we assess the legitimacy of earlier assumptions. Specifically, we show that the driving transport mechanisms for He and heat are of a fundamentally different nature for a high range of permeabilities ( k ≤ 10 - 16 m 2) found in metamorphic and volcanic rocks at all depths in the crust. The assumption that transport properties for these two tracers are similar in the crust is thus unsound. We also show that total 4He / heat flux ratios lower than radiogenic production ratios do not reflect a He deficit in the crust or mantle original reservoir. Instead, they

  15. Fuel cell with internal flow control

    SciTech Connect

    Haltiner, Jr., Karl J.; Venkiteswaran, Arun

    2012-06-12

    A fuel cell stack is provided with a plurality of fuel cell cassettes where each fuel cell cassette has a fuel cell with an anode and cathode. The fuel cell stack includes an anode supply chimney for supplying fuel to the anode of each fuel cell cassette, an anode return chimney for removing anode exhaust from the anode of each fuel cell cassette, a cathode supply chimney for supplying oxidant to the cathode of each fuel cell cassette, and a cathode return chimney for removing cathode exhaust from the cathode of each fuel cell cassette. A first fuel cell cassette includes a flow control member disposed between the anode supply chimney and the anode return chimney or between the cathode supply chimney and the cathode return chimney such that the flow control member provides a flow restriction different from at least one other fuel cell cassettes.

  16. 2D and 3D collagen and fibrin biopolymers promote specific ECM and integrin gene expression by vascular smooth muscle cells

    PubMed Central

    HONG, HELEN; STEGEMANN, JAN P.

    2009-01-01

    Collagen Type I and fibrin are polymeric proteins commonly used in the field of regenerative medicine as the foundational matrix of engineered tissues. We examined the response of vascular smooth muscle cells (VSMC) to both two-dimensional (2D) substrates as well as three-dimensional (3D) matrices of these biopolymers. Pure collagen Type I, pure fibrin and composite matrices consisting of 1:1 mixtures of collagen and fibrin were studied. Relative gene expression of three ECM molecules (collagen Type I and III, and tropoelastin) and three integrin subunits (integrins α1, β1 and β3) was determined over 7 days in culture using quantitative RT-PCR. Expression of all of these marker genes was up-regulated in 3D matrices, relative to 2D substrates. Tropoelastin, integrin α1 and integrin β1 were highest in collagen matrices, while collagen III and integrin β3 expression were highest in pure fibrin, and collagen I expression was highest in the collagen-fibrin composite materials. Both the compositional and temporal expression patterns of these specific ECM-related genes were suggestive of a wound healing response. These results illuminate the short-term responses of VSMC to 2D and 3D biopolymer matrices, and have relevance to tissue engineering and cardiovascular biology. PMID:18854122

  17. Verification and benchmarking of MAGNUM-2D: a finite element computer code for flow and heat transfer in fractured porous media

    SciTech Connect

    Eyler, L.L.; Budden, M.J.

    1985-03-01

    The objective of this work is to assess prediction capabilities and features of the MAGNUM-2D computer code in relation to its intended use in the Basalt Waste Isolation Project (BWIP). This objective is accomplished through a code verification and benchmarking task. Results are documented which support correctness of prediction capabilities in areas of intended model application. 10 references, 43 figures, 11 tables.

  18. ICEG2D: An Integrated Software Package for Automated Prediction of Flow Fields for Single-Element Airfoils with Ice Accretion

    NASA Technical Reports Server (NTRS)

    Thompson, David S.; Soni, Bharat K.

    2000-01-01

    An integrated software package, ICEG2D, was developed to automate computational fluid dynamics (CFD) simulations for single-element airfoils with ice accretion. ICEG2D is designed to automatically perform three primary functions: (1) generating a grid-ready, surface definition based on the geometrical characteristics of the iced airfoil surface, (2) generating a high-quality grid using the generated surface point distribution, and (3) generating the input and restart files needed to run the general purpose CFD solver NPARC. ICEG2D can be executed in batch mode using a script file or in an interactive mode by entering directives from a command line. This report summarizes activities completed in the first year of a three-year research and development program to address issues related to CFD simulations for aircraft components with ice accretion. Specifically, this document describes the technology employed in the software, the installation procedure, and a description of the operation of the software package. Validation of the geometry and grid generation modules of ICEG2D is also discussed.

  19. RGD-conjugated rod-like viral nanoparticles on 2D scaffold improved bone differentiation of mesenchymal stem cells

    NASA Astrophysics Data System (ADS)

    Wang, Qian; Pongkwan, Sitasuwan; Lee, L.; Li, Kai; Nguyen, Huong

    2014-05-01

    Viral nanoparticles have uniform and well-defined nano-structures and can be produced in large quantities. Several plant viral nanoparticles have been tested in biomedical applications due to the lack of mammalian cell infectivity. We are particularly interested in using Tobacco mosaic virus (TMV), which has been demonstrated to enhance bone tissue regeneration, as a tuneable nanoscale building block for biomaterials development. Unmodified TMV particles have been shown to accelerate osteogenic differentiation of adult stem cells by synergistically upregulating BMP2 and IBSP expression with dexamethasone. However, the lack of affinity to mammalian cell surface resulted in low initial cell adhesion. In this study, to increase cell binding capacity of TMV based material the chemical functionalization of TMV with arginine-glycine-aspartic acid (RGD) peptide was explored. An azide-derivatized RGD peptide was “clicked” to tyrosine residues on TMV outer surface via an efficient copper(I) catalysed azide-alkyne cycloaddition reaction. The ligand spacing is calculated to be 2-4 nm, which could offer a polyvalent ligand clustering effect for enhanced cell receptor signalling, further promoting the proliferation and osteogenic differentiation of bone marrow derived mesenchymal stem cells.

  20. Opportunities for Live Cell FT-Infrared Imaging: Macromolecule Identification with 2D and 3D Localization

    PubMed Central

    Mattson, Eric C.; Aboualizadeh, Ebrahim; Barabas, Marie E.; Stucky, Cheryl L.; Hirschmugl, Carol J.

    2013-01-01

    Infrared (IR) spectromicroscopy, or chemical imaging, is an evolving technique that is poised to make significant contributions in the fields of biology and medicine. Recent developments in sources, detectors, measurement techniques and speciman holders have now made diffraction-limited Fourier transform infrared (FTIR) imaging of cellular chemistry in living cells a reality. The availability of bright, broadband IR sources and large area, pixelated detectors facilitate live cell imaging, which requires rapid measurements using non-destructive probes. In this work, we review advances in the field of FTIR spectromicroscopy that have contributed to live-cell two and three-dimensional IR imaging, and discuss several key examples that highlight the utility of this technique for studying the structure and chemistry of living cells. PMID:24256815

  1. 31P NMR 2D Mapping of Creatine Kinase Forward Flux Rate in Hearts with Postinfarction Left Ventricular Remodeling in Response to Cell Therapy

    PubMed Central

    Gao, Ling; Cui, Weina; Zhang, Pengyuan; Jang, Albert; Zhu, Wuqiang; Zhang, Jianyi

    2016-01-01

    Utilizing a fast 31P magnetic resonance spectroscopy (MRS) 2-dimensional chemical shift imaging (2D-CSI) method, this study examined the heterogeneity of creatine kinase (CK) forward flux rate of hearts with postinfarction left ventricular (LV) remodeling. Immunosuppressed Yorkshire pigs were assigned to 4 groups: 1) A sham-operated normal group (SHAM, n = 6); 2) A 60 minutes distal left anterior descending coronary artery ligation and reperfusion (MI, n = 6); 3) Open patch group; ligation injury plus open fibrin patch over the site of injury (Patch, n = 6); and 4) Cell group, hiPSCs-cardiomyocytes, -endothelial cells, and -smooth muscle cells (2 million, each) were injected into the injured myocardium pass through a fibrin patch (Cell+Patch, n = 5). At 4 weeks, the creatine phosphate (PCr)/ATP ratio, CK forward flux rate (Flux PCr→ATP), and k constant of CK forward flux rate (kPCr→ATP) were severely decreased at border zone myocardium (BZ) adjacent to MI. Cell treatment results in significantly increase of PCr/ATP ratio and improve the value of kPCr→ATP and Flux PCr→ATP in BZ myocardium. Moreover, the BZ myocardial CK total activity and protein expression of CK mitochondria isozyme and CK myocardial isozyme were significantly reduced, but recovered in response to cell treatment. Thus, cell therapy results in improvement of BZ bioenergetic abnormality in hearts with postinfarction LV remodeling, which is accompanied by significantly improvements in BZ CK activity and CK isozyme expression. The fast 2D 31P MR CSI mapping can reliably measure the heterogeneity of bioenergetics in hearts with post infarction LV remodeling. PMID:27606901

  2. 31P NMR 2D Mapping of Creatine Kinase Forward Flux Rate in Hearts with Postinfarction Left Ventricular Remodeling in Response to Cell Therapy.

    PubMed

    Gao, Ling; Cui, Weina; Zhang, Pengyuan; Jang, Albert; Zhu, Wuqiang; Zhang, Jianyi

    2016-01-01

    Utilizing a fast 31P magnetic resonance spectroscopy (MRS) 2-dimensional chemical shift imaging (2D-CSI) method, this study examined the heterogeneity of creatine kinase (CK) forward flux rate of hearts with postinfarction left ventricular (LV) remodeling. Immunosuppressed Yorkshire pigs were assigned to 4 groups: 1) A sham-operated normal group (SHAM, n = 6); 2) A 60 minutes distal left anterior descending coronary artery ligation and reperfusion (MI, n = 6); 3) Open patch group; ligation injury plus open fibrin patch over the site of injury (Patch, n = 6); and 4) Cell group, hiPSCs-cardiomyocytes, -endothelial cells, and -smooth muscle cells (2 million, each) were injected into the injured myocardium pass through a fibrin patch (Cell+Patch, n = 5). At 4 weeks, the creatine phosphate (PCr)/ATP ratio, CK forward flux rate (Flux PCr→ATP), and k constant of CK forward flux rate (kPCr→ATP) were severely decreased at border zone myocardium (BZ) adjacent to MI. Cell treatment results in significantly increase of PCr/ATP ratio and improve the value of kPCr→ATP and Flux PCr→ATP in BZ myocardium. Moreover, the BZ myocardial CK total activity and protein expression of CK mitochondria isozyme and CK myocardial isozyme were significantly reduced, but recovered in response to cell treatment. Thus, cell therapy results in improvement of BZ bioenergetic abnormality in hearts with postinfarction LV remodeling, which is accompanied by significantly improvements in BZ CK activity and CK isozyme expression. The fast 2D 31P MR CSI mapping can reliably measure the heterogeneity of bioenergetics in hearts with post infarction LV remodeling. PMID:27606901

  3. Organic Solar Cells Based on a 2D Benzo[1,2-b:4,5-b']difuran-Conjugated Polymer with High-Power Conversion Efficiency.

    PubMed

    Huo, Lijun; Liu, Tao; Fan, Bingbing; Zhao, Zhiyuan; Sun, Xiaobo; Wei, Donghui; Yu, Mingming; Liu, Yunqi; Sun, Yanming

    2015-11-18

    A novel 2D benzodifuran (BDF)-based copolymer (PBDF-T1) is synthesized. Polymer solar cells fabricated with PBDF-T1 show high power conversion efficiency of 9.43% and fill factor of 77.4%, which is higher than the performance of its benzothiophene (BDT) counterpart (PBDT-T1). These results provide important progress for BDF-based copolymers and demonstrate that BDF-based copolymers can be competitive with the well-studied BDT counterparts via molecular structure design and device optimization.

  4. Blood Cell Interactions and Segregation in Flow

    PubMed Central

    Munn, Lance L.; Dupin, Michael M.

    2009-01-01

    For more than a century, pioneering researchers have been using novel experimental and computational approaches to probe the mysteries of blood flow. Thanks to their efforts, we know that blood cells generally prefer to migrate to the axis of flow, that red and white cells segregate in flow, and that cell deformability and their tendency to reversibly aggregate contribute to the non-Newtonian nature of this unique fluid. All of these properties have beneficial physiological consequences, allowing blood to perform a variety of critical functions. Our current understanding of these unusual flow properties of blood have been made possible by the ingenuity and diligence of a number of researchers, including Harry Goldsmith, who developed novel technologies to visualize and quantify the flow of blood at the level of individual cells. Here we summarize efforts in our lab to continue this tradition and to further our understanding of how blood cells interact with each other and with the blood vessel wall. PMID:18188702

  5. A bispecific protein rG7S-MICA recruits natural killer cells and enhances NKG2D-mediated immunosurveillance against hepatocellular carcinoma.

    PubMed

    Wang, Tong; Sun, Fumou; Xie, Wei; Tang, Mingying; He, Hua; Jia, Xuelian; Tian, Xuemei; Wang, Min; Zhang, Juan

    2016-03-28

    MHC class I-related chain A (MICA) is a principal immunoligand of the natural killer (NK) cell receptor NK group 2, member D (NKG2D) and plays a key role in NK cell-mediated immune recognition. Shedding of MICA from tumor cells leads to immunosuppression. To reconstitute the immunosurveilance function of NK cells, we constructed a fusion protein rG7S-MICA and explored its potential anti-tumor activity against hepatocellular carcinoma (HCC). rG7S-MICA consists of human MICA and a single-chain antibody fragment (scFv) targeting the tumor-associated antigen cluster of differentiation 24 (CD24). In vitro, rG7S-MICA engaged both NK cells and CD24(+) human HCC cells, and triggered NK cell-mediated cytolysis. Furthermore, in CD24(+) HCC-bearing nude mice, rG7S-MICA specifically targeted to the tumor tissue, where it effectively recruited NK cells and induced the release of cytokines, and showed superior anti-tumor activity. In conclusion, rG7S-MICA provides a new approach for HCC-targeting immunotherapy and has attracting potentials for clinical applications. PMID:26791237

  6. A bispecific protein rG7S-MICA recruits natural killer cells and enhances NKG2D-mediated immunosurveillance against hepatocellular carcinoma.

    PubMed

    Wang, Tong; Sun, Fumou; Xie, Wei; Tang, Mingying; He, Hua; Jia, Xuelian; Tian, Xuemei; Wang, Min; Zhang, Juan

    2016-03-28

    MHC class I-related chain A (MICA) is a principal immunoligand of the natural killer (NK) cell receptor NK group 2, member D (NKG2D) and plays a key role in NK cell-mediated immune recognition. Shedding of MICA from tumor cells leads to immunosuppression. To reconstitute the immunosurveilance function of NK cells, we constructed a fusion protein rG7S-MICA and explored its potential anti-tumor activity against hepatocellular carcinoma (HCC). rG7S-MICA consists of human MICA and a single-chain antibody fragment (scFv) targeting the tumor-associated antigen cluster of differentiation 24 (CD24). In vitro, rG7S-MICA engaged both NK cells and CD24(+) human HCC cells, and triggered NK cell-mediated cytolysis. Furthermore, in CD24(+) HCC-bearing nude mice, rG7S-MICA specifically targeted to the tumor tissue, where it effectively recruited NK cells and induced the release of cytokines, and showed superior anti-tumor activity. In conclusion, rG7S-MICA provides a new approach for HCC-targeting immunotherapy and has attracting potentials for clinical applications.

  7. Interactions of 1D- and 2D-layered inorganic nanoparticles with fibroblasts and human mesenchymal stem cells

    PubMed Central

    Rashkow, Jason Thomas; Talukdar, Yahfi; Lalwani, Gaurav; Sitharaman, Balaji

    2015-01-01

    Aim This study investigates the effects of tungsten disulfide nanotubes (WSNTs) and molybdenum disulfide nanoplatelets (MSNPs) on fibroblasts (NIH-3T3) and mesenchymal stem cells (MSCs) to determine safe dosages for potential biomedical applications. Materials & methods Cytotoxicity of MSNPs and WSNTs (5–300 µg/ml) on NIH-3T3 and MSCs was assessed at 6, 12 or 24 h. MSC differentiation to adipocytes and osteoblasts was assessed following treatment for 24 h. Results Only NIH-3T3 cells treated with MSNPs showed dose or time dependent increase in cytotoxicity. Differentiation markers of MSCs in treated groups were unaffected compared with untreated controls. Conclusion MSNPs and WSNTs at concentrations less than 50 µg/ml are potentially safe for treatment of fibroblasts or MSCs for up to 24 h. PMID:26080694

  8. Spatio-temporal morphology changes in and quenching effects on the 2D spreading dynamics of cell colonies in both plain and methylcellulose-containing culture media.

    PubMed

    Muzzio, N E; Pasquale, M A; Huergo, M A C; Bolzán, A E; González, P H; Arvia, A J

    2016-06-01

    To deal with complex systems, microscopic and global approaches become of particular interest. Our previous results from the dynamics of large cell colonies indicated that their 2D front roughness dynamics is compatible with the standard Kardar-Parisi-Zhang (KPZ) or the quenched KPZ equations either in plain or methylcellulose (MC)-containing gel culture media, respectively. In both cases, the influence of a non-uniform distribution of the colony constituents was significant. These results encouraged us to investigate the overall dynamics of those systems considering the morphology and size, the duplication rate, and the motility of single cells. For this purpose, colonies with different cell populations (N) exhibiting quasi-circular and quasi-linear growth fronts in plain and MC-containing culture media are investigated. For small N, the average radial front velocity and its change with time depend on MC concentration. MC in the medium interferes with cell mitosis, contributes to the local enlargement of cells, and increases the distribution of spatio-temporal cell density heterogeneities. Colony spreading in MC-containing media proceeds under two main quenching effects, I and II; the former mainly depending on the culture medium composition and structure and the latter caused by the distribution of enlarged local cell domains. For large N, colony spreading occurs at constant velocity. The characteristics of cell motility, assessed by measuring their trajectories and the corresponding velocity field, reflect the effect of enlarged, slow-moving cells and the structure of the medium. Local average cell size distribution and individual cell motility data from plain and MC-containing media are qualitatively consistent with the predictions of both the extended cellular Potts models and the observed transition of the front roughness dynamics from a standard KPZ to a quenched KPZ. In this case, quenching effects I and II cooperate and give rise to the quenched

  9. Graphene Paper Decorated with a 2D Array of Dendritic Platinum Nanoparticles for Ultrasensitive Electrochemical Detection of Dopamine Secreted by Live Cells.

    PubMed

    Zan, Xiaoli; Bai, Hongwei; Wang, Chenxu; Zhao, Faqiong; Duan, Hongwei

    2016-04-01

    To circumvent the bottlenecks of non-flexibility, low sensitivity, and narrow workable detection range of conventional biosensors for biological molecule detection (e.g., dopamine (DA) secreted by living cells), a new hybrid flexible electrochemical biosensor has been created by decorating closely packed dendritic Pt nanoparticles (NPs) on freestanding graphene paper. This innovative structural integration of ultrathin graphene paper and uniform 2D arrays of dendritic NPs by tailored wet chemical synthesis has been achieved by a modular strategy through a facile and delicately controlled oil-water interfacial assembly method, whereby the uniform distribution of catalytic dendritic NPs on the graphene paper is maximized. In this way, the performance is improved by several orders of magnitude. The developed hybrid electrode shows a high sensitivity of 2 μA cm(-2) μM(-1), up to about 33 times higher than those of conventional sensors, a low detection limit of 5 nM, and a wide linear range of 87 nM to 100 μM. These combined features enable the ultrasensitive detection of DA released from pheochromocytoma (PC 12) cells. The unique features of this flexible sensor can be attributed to the well-tailored uniform 2D array of dendritic Pt NPs and the modular electrode assembly at the oil-water interface. Its excellent performance holds much promise for the future development of optimized flexible electrochemical sensors for a diverse range of electroactive molecules to better serve society. PMID:26918612

  10. Graphene Paper Decorated with a 2D Array of Dendritic Platinum Nanoparticles for Ultrasensitive Electrochemical Detection of Dopamine Secreted by Live Cells.

    PubMed

    Zan, Xiaoli; Bai, Hongwei; Wang, Chenxu; Zhao, Faqiong; Duan, Hongwei

    2016-04-01

    To circumvent the bottlenecks of non-flexibility, low sensitivity, and narrow workable detection range of conventional biosensors for biological molecule detection (e.g., dopamine (DA) secreted by living cells), a new hybrid flexible electrochemical biosensor has been created by decorating closely packed dendritic Pt nanoparticles (NPs) on freestanding graphene paper. This innovative structural integration of ultrathin graphene paper and uniform 2D arrays of dendritic NPs by tailored wet chemical synthesis has been achieved by a modular strategy through a facile and delicately controlled oil-water interfacial assembly method, whereby the uniform distribution of catalytic dendritic NPs on the graphene paper is maximized. In this way, the performance is improved by several orders of magnitude. The developed hybrid electrode shows a high sensitivity of 2 μA cm(-2) μM(-1), up to about 33 times higher than those of conventional sensors, a low detection limit of 5 nM, and a wide linear range of 87 nM to 100 μM. These combined features enable the ultrasensitive detection of DA released from pheochromocytoma (PC 12) cells. The unique features of this flexible sensor can be attributed to the well-tailored uniform 2D array of dendritic Pt NPs and the modular electrode assembly at the oil-water interface. Its excellent performance holds much promise for the future development of optimized flexible electrochemical sensors for a diverse range of electroactive molecules to better serve society.

  11. Plasmonic spectrum on 1D and 2D periodic arrays of rod-shape metal nanoparticle pairs with different core patterns for biosensor and solar cell applications

    NASA Astrophysics Data System (ADS)

    Kumara, N. T. R. N.; Chou Chau, Yuan-Fong; Huang, Jin-Wei; Huang, Hung Ji; Lin, Chun-Ting; Chiang, Hai-Pang

    2016-11-01

    Simulations of surface plasmon resonance (SPR) on the near field intensity and absorption spectra of one-dimensional (1D) and two-dimensional (2D) periodic arrays of rod-shape metal nanoparticle (MNP) pairs using the finite element method (FEM) and taking into account the different core patterns for biosensor and solar cell applications are investigated. A tunable optical spectrum corresponding to the transverse SPR modes is observed. The peak resonance wavelength (λ res) can be shifted to red as the core patterns in rod-shape MNPs have been changed. We find that the 2D periodic array of core–shell MNP pairs (case 2) exhibit a red shifted SPR that can be tuned the gap enhancement and absorption efficiency simultaneously over an extended wavelength range. The tunable optical performances give us a qualitative idea of the geometrical properties of the periodic array of rod-shape MNP pairs on SPRs that can be as a promising candidate for plasmonic biosensor and solar cell applications.

  12. Graphene Paper Decorated with a 2D Array of Dendritic Platinum Nanoparticles for Ultrasensitive Electrochemical Detection of Dopamine Secreted by Live Cells

    PubMed Central

    Zan, Xiaoli; Wang, Chenxu

    2016-01-01

    Abstract To circumvent the bottlenecks of non‐flexibility, low sensitivity, and narrow workable detection range of conventional biosensors for biological molecule detection (e.g., dopamine (DA) secreted by living cells), a new hybrid flexible electrochemical biosensor has been created by decorating closely packed dendritic Pt nanoparticles (NPs) on freestanding graphene paper. This innovative structural integration of ultrathin graphene paper and uniform 2D arrays of dendritic NPs by tailored wet chemical synthesis has been achieved by a modular strategy through a facile and delicately controlled oil–water interfacial assembly method, whereby the uniform distribution of catalytic dendritic NPs on the graphene paper is maximized. In this way, the performance is improved by several orders of magnitude. The developed hybrid electrode shows a high sensitivity of 2 μA cm−2 μm −1, up to about 33 times higher than those of conventional sensors, a low detection limit of 5 nm, and a wide linear range of 87 nm to 100 μm. These combined features enable the ultrasensitive detection of DA released from pheochromocytoma (PC 12) cells. The unique features of this flexible sensor can be attributed to the well‐tailored uniform 2D array of dendritic Pt NPs and the modular electrode assembly at the oil–water interface. Its excellent performance holds much promise for the future development of optimized flexible electrochemical sensors for a diverse range of electroactive molecules to better serve society. PMID:26918612

  13. 1,25(OH)2D3 attenuates TGF-β1/β2-induced increased migration and invasion via inhibiting epithelial-mesenchymal transition in colon cancer cells.

    PubMed

    Chen, Shanwen; Zhu, Jing; Zuo, Shuai; Ma, Ju; Zhang, Junling; Chen, Guowei; Wang, Xin; Pan, Yisheng; Liu, Yucun; Wang, Pengyuan

    1,25-Dihydroxyvitamin D3 (1,25(OH)2D3) has been reported to inhibit proliferation and migration of multiple types of cancer cells. However, the mechanism underlying its anti-metastasis effect is not fully illustrated. In this study, the effect of 1,25(OH)2D3 on TGF-β1/β2-induced epithelial-mesenchymal transition (EMT) is tested in colon cancer cells. The results suggest that 1,25(OH)2D3 inhibited TGF-β1/β2-induced increased invasion and migration of in SW-480 and HT-29 cells. 1,25(OH)2D3 also inhibited the cadherin switch in SW-480 and HT-29 cells. TGF-β1/β2-induced increased expression of EMT-related transcription factors was also inhibited by 1,25(OH)2D3. 1,25(OH)2D3 also inhibited the secretion of MMP-2 and MMP-9 and increased expression of F-actin induced by TGF-β1/β2 in SW-480 cells. Taken together, this study suggests that the suppression of EMT might be one of the mechanisms underlying the anti-metastasis effect of 1,25(OH)2D3 in colon cancer cells.

  14. Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry.

    PubMed

    Davidenko, Natalia; Schuster, Carlos F; Bax, Daniel V; Farndale, Richard W; Hamaia, Samir; Best, Serena M; Cameron, Ruth E

    2016-10-01

    Studies of cell attachment to collagen-based materials often ignore details of the binding mechanisms-be they integrin-mediated or non-specific. In this work, we have used collagen and gelatin-based substrates with different dimensional characteristics (monolayers, thin films and porous scaffolds) in order to establish the influence of composition, crosslinking (using carbodiimide) treatment and 2D or 3D architecture on integrin-mediated cell adhesion. By varying receptor expression, using cells with collagen-binding integrins (HT1080 and C2C12 L3 cell lines, expressing α2β1, and Rugli expressing α1β1) and a parent cell line C2C12 with gelatin-binding receptors (αvβ3 and α5β1), the nature of integrin binding sites was studied in order to explain the bioactivity of different protein formulations. We have shown that alteration of the chemical identity, conformation and availability of free binding motifs (GxOGER and RGD), resulting from addition of gelatin to collagen and crosslinking, have a profound effect on the ability of cells to adhere to these formulations. Carbodiimide crosslinking ablates integrin-dependent cell activity on both two-dimensional and three-dimensional architectures while the three-dimensional scaffold structure also leads to a high level of non-specific interactions remaining on three-dimensional samples even after a rigorous washing regime. This phenomenon, promoted by crosslinking, and attributed to cell entrapment, should be considered in any assessment of the biological activity of three-dimensional substrates. Spreading data confirm the importance of integrin-mediated cell engagement for further cell activity on collagen-based compositions. In this work, we provide a simple, but effective, means of deconvoluting the effects of chemistry and dimensional characteristics of a substrate, on the cell activity of protein-derived materials, which should assist in tailoring their biological properties for specific tissue engineering

  15. Evaluation of cell binding to collagen and gelatin: a study of the effect of 2D and 3D architecture and surface chemistry.

    PubMed

    Davidenko, Natalia; Schuster, Carlos F; Bax, Daniel V; Farndale, Richard W; Hamaia, Samir; Best, Serena M; Cameron, Ruth E

    2016-10-01

    Studies of cell attachment to collagen-based materials often ignore details of the binding mechanisms-be they integrin-mediated or non-specific. In this work, we have used collagen and gelatin-based substrates with different dimensional characteristics (monolayers, thin films and porous scaffolds) in order to establish the influence of composition, crosslinking (using carbodiimide) treatment and 2D or 3D architecture on integrin-mediated cell adhesion. By varying receptor expression, using cells with collagen-binding integrins (HT1080 and C2C12 L3 cell lines, expressing α2β1, and Rugli expressing α1β1) and a parent cell line C2C12 with gelatin-binding receptors (αvβ3 and α5β1), the nature of integrin binding sites was studied in order to explain the bioactivity of different protein formulations. We have shown that alteration of the chemical identity, conformation and availability of free binding motifs (GxOGER and RGD), resulting from addition of gelatin to collagen and crosslinking, have a profound effect on the ability of cells to adhere to these formulations. Carbodiimide crosslinking ablates integrin-dependent cell activity on both two-dimensional and three-dimensional architectures while the three-dimensional scaffold structure also leads to a high level of non-specific interactions remaining on three-dimensional samples even after a rigorous washing regime. This phenomenon, promoted by crosslinking, and attributed to cell entrapment, should be considered in any assessment of the biological activity of three-dimensional substrates. Spreading data confirm the importance of integrin-mediated cell engagement for further cell activity on collagen-based compositions. In this work, we provide a simple, but effective, means of deconvoluting the effects of chemistry and dimensional characteristics of a substrate, on the cell activity of protein-derived materials, which should assist in tailoring their biological properties for specific tissue engineering

  16. Aniso2D

    2005-07-01

    Aniso2d is a two-dimensional seismic forward modeling code. The earth is parameterized by an X-Z plane in which the seismic properties Can have monoclinic with x-z plane symmetry. The program uses a user define time-domain wavelet to produce synthetic seismograms anrwhere within the two-dimensional media.

  17. Towards 2D nanocomposites

    NASA Astrophysics Data System (ADS)

    Jang, Hyun-Sook; Yu, Changqian; Hayes, Robert; Granick, Steve

    2015-03-01

    Polymer vesicles (``polymersomes'') are an intriguing class of soft materials, commonly used to encapsulate small molecules or particles. Here we reveal they can also effectively incorporate nanoparticles inside their polymer membrane, leading to novel ``2D nanocomposites.'' The embedded nanoparticles alter the capacity of the polymersomes to bend and to stretch upon external stimuli.

  18. 2D-DIGE screening of high-productive CHO cells under glucose limitation--basic changes in the proteome equipment and hints for epigenetic effects.

    PubMed

    Wingens, Marc; Gätgens, Jochem; Schmidt, Anica; Albaum, Stefan P; Büntemeyer, Heino; Noll, Thomas; Hoffrogge, Raimund

    2015-05-10

    CHO derivates (Chinese hamster ovary) belong to the most important mammalian cells for industrial recombinant protein production. Many efforts have been made to improve productivity and stability of CHO cells in bioreactor processes. Here, we followed up one barely understood phenomenon observed with process optimizations: a significantly increased cell-specific productivity in late phases of glucose-limited perfusion cultivations, when glucose (and lactate) reserves are exhausted. Our aim was to elucidate the cellular activities connected to the metabolic shift from glucose surplus to glucose limitation phase. With 2D-DIGE, we compared three stages in a perfusion culture of CHO cells: the initial growth with high glucose concentration and low lactate production, the second phase with glucose going to limitation and high lactate level, and finally the state of glucose limitation and also low lactate concentration but increased cell-specific productivity. With our proteomic approach we were able to demonstrate consequences of glucose limitation for the protein expression machinery which also could play a role for a higher recombinant protein production. Most interestingly, we detected epigenetic effects on the level of proteins involved in histone modification (HDAC1/-2, SET, RBBP7, DDX5). Together with shifts in the protein inventory of energy metabolism, cytoskeleton and protein expression, a picture emerges of basic changes in the cellular equipment under long-term glucose limitation of CHO cells.

  19. Solution-processed 2D niobium diselenide nanosheets as efficient hole-transport layers in organic solar cells.

    PubMed

    Gu, Xing; Cui, Wei; Song, Tao; Liu, Changhai; Shi, Xiaoze; Wang, Suidong; Sun, Baoquan

    2014-02-01

    Thin-layer, two-dimensional NbSe2 nanosheets with lower trap density have been obtained and act as an alternative hole-transporting layer to replace MoO3 in organic solar cells. If poly({4,8-bis[(2-ethylhexyl)oxy]benzo[1,2-b:4,5-b']dithiophene-2,6-diyl}{3-fluoro-2-[(2-ethylhexyl)carbonyl]thieno[3,4-b]thiophenediyl}):[6,6]-phenyl-C71-butyric acid methyl ester acts as an active layer, a power conversion efficiency of 8.10 % has been achieved without any further thermal treatment. The properties of this hole-transporting layer were investigated and the improvements in the devices are discussed.

  20. Performance Mapping Studies in Redox Flow Cells

    NASA Technical Reports Server (NTRS)

    Hoberecht, M. A.; Thaller, L. H.

    1981-01-01

    Pumping power requirements in any flow battery system constitute a direct parasitic energy loss. It is therefore useful to determine the practical lower limit for reactant flow rates. Through the use of a theoretical framework based on electrochemical first principles, two different experimental flow mapping techniques were developed to evaluate and compare electrodes as a function of flow rate. For the carbon felt electrodes presently used in NASA-Lewis Redox cells, a flow rate 1.5 times greater than the stoichiometric rate seems to be the required minimum.

  1. Differential response to 1α, 25-dihydroxyvitamin D3 (1α,25(OH)2D3) in non-small cell lung cancer cells with distinct oncogene mutations1

    PubMed Central

    Zhang, Qiuhong; Kanterewicz, Beatriz; Shoemaker, Suzanne; Hu, Qiang; Liu, Song; Atwood, Kristopher; Hershberger, Pamela

    2012-01-01

    We previously demonstrated that non-small cell lung cancer (NSCLC) cells and primary human lung tumors aberrantly express the vitamin D3-catabolizing enzyme, CYP24, and that CYP24 restricts transcriptional regulation and growth control by 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) in NSCLC cells. To ascertain the basis for CYP24 dysregulation, we assembled a panel of cell lines that represent distinct molecular classes of lung cancer: Cell lines were selected which harbored mutually exclusive mutations in either the K-ras or the Epidermal Growth Factor Receptor (EGFR) genes. We observed that K-ras mutant lines displayed a basal vitamin D receptor (VDR)lowCYP24high phenotype, whereas EGFR mutant lines had a VDRhighCYP24low phenotype. A mutation-associated difference in CYP24 expression was also observed in clinical specimens. Specifically, K-ras mutation was associated with a median 4.2-fold increase in CYP24 mRNA expression (p = 4.8 × 10−7) compared to EGFR mutation in a series of 147 primary lung adenocarcinoma cases. Because of their differential basal expression of VDR and CYP24, we hypothesized that NSCLC cells with an EGFR mutation would be more responsive to 1,25(OH)2D3 treatment than those with a K-ras mutation. To test this, we measured the ability of 1,25(OH)2D3 to increase reporter gene activity, induce transcription of endogenous target genes, and suppress colony formation. In each assay, the extent of 1,25(OH)2D3 response was greater in EGFR mutation-positive HCC827 and H1975 cells than in K-ras mutation-positive A549 and 128.88T cells. We subsequently examined the effect of combining 1,25(OH)2D3 with erlotinib, which is used clinically in the treatment of EGFR mutation-positive NSCLC. 1,25(OH)2D3/erlotinib combination resulted in significantly greater growth inhibition than either single agent in both the erlotinib-sensitive HCC827 cell line and the erlotinib-resistant H1975 cell line. These data are the first to suggest that EGFR mutations may

  2. Rapid 2D incoherent mirror fabrication by laser interference lithography and wet etching for III-V MQW solar cells

    NASA Astrophysics Data System (ADS)

    Wang, Wei; Freundlich, Alex

    2016-03-01

    Optimization of non-planar antireflective coating and back- (or front-) surface texturing are widely studied as advanced light management approach to further reduce the reflection losses and increase the sunlight absorption path in solar cells. Rear reflectors have been developed from coherent mirrors to incoherent mirrors in order to further increase light path, which can significantly improve the efficiency and allow for much thinner devices. A Lambertian surface, which has the most random texture, can theoretically raise the light path to 4n2 times that of a smooth surface. It's a challenge however to fabricate ideal Lambertian texture, especially in a fast and low cost way. In this work, a method is developed to overcome this challenge that combines the use of laser interference lithography (LIL) and selective wet etching. This approach allows for a rapid (10 min) wafer scale (3 inch wafer) texture processing with sub-wavelength (nano)-scale control of the pattern and the pitch. The technique appears as being particularly attractive for the development of ultrathin III-V devices, or in overcoming the weak sub-bandgap absorption in devices incorporating quantum dots or quantum wells. The structure of the device is demonstrated, without affecting active layers.

  3. A New Cell-Centered Implicit Numerical Scheme for Ions in the 2-D Axisymmetric Code Hall2de

    NASA Technical Reports Server (NTRS)

    Lopez Ortega, Alejandro; Mikellides, Ioannis G.

    2014-01-01

    We present a new algorithm in the Hall2De code to simulate the ion hydrodynamics in the acceleration channel and near plume regions of Hall-effect thrusters. This implementation constitutes an upgrade of the capabilities built in the Hall2De code. The equations of mass conservation and momentum for unmagnetized ions are solved using a conservative, finite-volume, cell-centered scheme on a magnetic-field-aligned grid. Major computational savings are achieved by making use of an implicit predictor/multi-corrector algorithm for time evolution. Inaccuracies in the prediction of the motion of low-energy ions in the near plume in hydrodynamics approaches are addressed by implementing a multi-fluid algorithm that tracks ions of different energies separately. A wide range of comparisons with measurements are performed to validate the new ion algorithms. Several numerical experiments with the location and value of the anomalous collision frequency are also presented. Differences in the plasma properties in the near-plume between the single fluid and multi-fluid approaches are discussed. We complete our validation by comparing predicted erosion rates at the channel walls of the thruster with measurements. Erosion rates predicted by the plasma properties obtained from simulations replicate accurately measured rates of erosion within the uncertainty range of the sputtering models employed.

  4. Application of rank-ordered multifractal analysis (ROMA) to intermittent fluctuations in 3D turbulent flows, 2D MHD simulation and solar wind data

    NASA Astrophysics Data System (ADS)

    Wu, C.; Chang, T.

    2010-12-01

    A new method in describing the multifractal characteristics of intermittent events was introduced by Cheng and Wu [Chang T. and Wu C.C., Physical Rev, E77, 045401(R), 2008]. The procedure provides a natural connection between the rank-ordered spectrum and the idea of one-parameter scaling for monofractals. This technique has been demonstrated using results obtained from a 2D MHD simulation. It has also been successfully applied to in-situ solar wind observations [Chang T., Wu, C.C. and Podesta, J., AIP Conf Proc. 1039, 75, 2008], and the broadband electric field oscillations from the auroral zone [Tam, S.W.Y. et al., Physical Rev, E81, 036414, 2010]. We take the next step in this procedure. By using the ROMA spectra and the scaled probability distribution functions (PDFs), raw PDFs can be calculated, which can be compared directly with PDFs from observations or simulation results. In addition to 2D MHD simulation results and in-situ solar wind observation, we show clearly using the ROMA analysis the multifractal character of the 3D fluid simulation data obtained from the JHU turbulence database cluster at http://turbulence.pha.jhu.edu. In particular, we show the scaling of the non-symmetrical PDF for the parallel-velocity fluctuations of this 3D fluid data.

  5. An optical technique for the direct measurement of the 2-D spectral density of a passive scalar in a turbulent flow

    SciTech Connect

    Robey, H.F.; Albrecht, G.F.; Moore, T.R.

    1990-04-06

    A new optical technique for quantitatively measuring the spectral density of passive scalar fluctuations in a turbulent flow has been developed. The technique exploits the photorefractive properties of BaTiO{sub 3} to separate the optical signal of the turbulent field from the coherent illumination background. It is a major improvement over existing techniques in that it is non-intrusive, has excellent frequency response and spatial resolution, and is capable of simultaneously measuring two components of the three-dimensional spectral density, {Phi}{theta}({kappa}). The technique is thus especially well suited to the directly study of anisotropic flows. We have applied this technique to study the spectrum of temperature fluctuations in a fully developed turbulent channel flow with heat addition. The flow is highly anisotropic, yet the spectrum in directions transverse to the flow is seen to exhibit an inertial--convective subrange behavior which is characteristic of isotropic flows. The spectral behavior in the flow direction, due to the direct influence of the mean strain rate, is observed to be markedly different. 17 refs., 7 figs.

  6. Overhead Projection Cell for Streamline Flow

    ERIC Educational Resources Information Center

    Waage, Harold M.

    1969-01-01

    Describes the construction and operation of an overhead projection apparatus designed to demonstrate streamline flow of a liquid. The apparatus consists of a Plexiglass tank containing water in which plates forming the cell are submerged, a constant level reservoir, an overflow device and a system for marking the flow lines with a dye. (LC)

  7. Flow regimes in a trapped vortex cell

    NASA Astrophysics Data System (ADS)

    Lasagna, D.; Iuso, G.

    2016-03-01

    This paper presents results of an experimental investigation on the flow in a trapped vortex cell, embedded into a flat plate, and interacting with a zero-pressure-gradient boundary layer. The objective of the work is to describe the flow features and elucidate some of the governing physical mechanisms, in the light of recent investigations on flow separation control using vortex cells. Hot-wire velocity measurements of the shear layer bounding the cell and of the boundary layers upstream and downstream are reported, together with spectral and correlation analyses of wall-pressure fluctuation measurements. Smoke flow visualisations provide qualitative insight into some relevant features of the internal flow, namely a large-scale flow unsteadiness and possible mechanisms driving the rotation of the vortex core. Results are presented for two very different regimes: a low-Reynolds-number case where the incoming boundary layer is laminar and its momentum thickness is small compared to the cell opening, and a moderately high-Reynolds-number case, where the incoming boundary layer is turbulent and the ratio between the momentum thickness and the opening length is significantly larger than in the first case. Implications of the present findings to flow control applications of trapped vortex cells are also discussed.

  8. Ontology analysis of global gene expression differences of human bone marrow stromal cells cultured on 3D scaffolds or 2D films.

    PubMed

    Baker, Bryan A; Pine, P Scott; Chatterjee, Kaushik; Kumar, Girish; Lin, Nancy J; McDaniel, Jennifer H; Salit, Marc L; Simon, Carl G

    2014-08-01

    Differences in gene expression of human bone marrow stromal cells (hBMSCs) during culture in three-dimensional (3D) nanofiber scaffolds or on two-dimensional (2D) films were investigated via pathway analysis of microarray mRNA expression profiles. Previous work has shown that hBMSC culture in nanofiber scaffolds can induce osteogenic differentiation in the absence of osteogenic supplements (OS). Analysis using ontology databases revealed that nanofibers and OS regulated similar pathways and that both were enriched for TGF-β and cell-adhesion/ECM-receptor pathways. The most notable difference between the two was that nanofibers had stronger enrichment for cell-adhesion/ECM-receptor pathways. Comparison of nanofibers scaffolds with flat films yielded stronger differences in gene expression than comparison of nanofibers made from different polymers, suggesting that substrate structure had stronger effects on cell function than substrate polymer composition. These results demonstrate that physical (nanofibers) and biochemical (OS) signals regulate similar ontological pathways, suggesting that these cues use similar molecular mechanisms to control hBMSC differentiation. PMID:24840613

  9. Vector-averaged gravity-induced changes in cell signaling and vitamin D receptor activity in MG-63 cells are reversed by a 1,25-(OH)2D3 analog, EB1089

    NASA Technical Reports Server (NTRS)

    Narayanan, R.; Smith, C. L.; Weigel, N. L.

    2002-01-01

    Skeletal unloading in an animal hindlimb suspension model and microgravity experienced by astronauts or as a result of prolonged bed rest causes site-specific losses in bone mineral density of 1%-2% per month. This is accompanied by reductions in circulating levels of 1,25-(OH)(2)D(3), the active metabolite of vitamin D. 1,25-(OH)(2)D(3), the ligand for the vitamin D receptor (VDR), is important for calcium absorption and plays a role in differentiation of osteoblasts and osteoclasts. To examine the responses of cells to activators of the VDR in a simulated microgravity environment, we used slow-turning lateral vessels (STLVs) in a rotating cell culture system. We found that, similar to cells grown in microgravity, MG-63 cells grown in the STLVs produce less osteocalcin, alkaline phosphatase, and collagen Ialpha1 mRNA and are less responsive to 1,25-(OH)(2)D(3). In addition, expression of VDR was reduced. Moreover, growth in the STLV caused activation of the stress-activated protein kinase pathway (SAPK), a kinase that inhibits VDR activity. In contrast, the 1,25-(OH)(2)D(3) analog, EB1089, was able to compensate for some of the STLV-associated responses by reducing SAPK activity, elevating VDR levels, and increasing expression of osteocalcin and alkaline phosphatase. These studies suggest that, not only does simulated microgravity reduce differentiation of MG-63 cells, but the activity of the VDR, an important regulator of bone metabolism, is reduced. Use of potent, less calcemic analogs of 1,25-(OH)(2)D(3) may aid in overcoming this defect. Copyright 2002 Elsevier Science Inc.

  10. Solution-state 2D NMR of ball-milled plant cell wall gels in DMSO-d6/pyridine-d5†

    PubMed Central

    Ralph, John

    2014-01-01

    NMR fingerprinting of the components of finely divided plant cell walls swelled in DMSO has been recently described. Cell wall gels, produced directly in the NMR tube with perdeutero-dimethylsulfoxide, allowed the acquisition of well resolved/dispersed 2D 13C–1H correlated solution-state NMR spectra of the entire array of wall polymers, without the need for component fractionation. That is, without actual solubilization, and without apparent structural modification beyond that inflicted by the ball milling and ultrasonication steps, satisfactorily interpretable spectra can be acquired that reveal compositional and structural details regarding the polysaccharide and lignin components in the wall. Here, the profiling method has been improved by using a mixture of perdeuterated DMSO and pyridine (4:1, v/v). Adding pyridine provided not only easier sample handling because of the better mobility compared to the DMSO-d6-only system but also considerably elevated intensities and improved resolution of the NMR spectra due to the enhanced swelling of the cell walls. This modification therefore provides a more rapid method for comparative structural evaluation of plant cell walls than is currently available. We examined loblolly pine (Pinus taeda, a gymnosperm), aspen (Populus tremuloides, an angiosperm), kenaf (Hibiscus cannabinus, an herbaceous plant), and corn (Zea mays L., a grass, i.e., from the Poaceae family). In principle, lignin composition (notably, the syringyl : guaiacyl : p-hydroxyphenyl ratio) can be quantified without the need for lignin isolation. Correlations for p-coumarate units in the corn sample are readily seen, and a variety of the ferulate correlations are also well resolved; ferulates are important components responsible for cell wall cross-linking in grasses. Polysaccharide anomeric correlations were tentatively assigned for each plant sample based on standard samples and various literature data. With the new potential for chemometric analysis

  11. Quantifying the flow dynamics of supercritical CO2-water displacement in a 2D porous micromodel using fluorescent microscopy and microscopic PIV

    NASA Astrophysics Data System (ADS)

    Kazemifar, Farzan; Blois, Gianluca; Kyritsis, Dimitrios C.; Christensen, Kenneth T.

    2016-09-01

    The multi-phase flow of liquid/supercritical CO2 and water (non-wetting and wetting phases, respectively) in a two-dimensional silicon micromodel was investigated at reservoir conditions (80 bar, 24 °C and 40 °C). The fluorescent microscopy and microscopic particle image velocimetry (micro-PIV) techniques were combined to quantify the flow dynamics associated with displacement of water by CO2 (drainage) in the porous matrix. To this end, water was seeded with fluorescent tracer particles, CO2 was tagged with a fluorescent dye and each phase was imaged independently using spectral separation in conjunction with microscopic imaging. This approach allowed simultaneous measurement of the spatially-resolved instantaneous velocity field in the water and quantification of the spatial configuration of the two fluid phases. The results, acquired with sufficient time resolution to follow the dynamic progression of both phases, provide a comprehensive picture of the flow physics during the migration of the CO2 front, the temporal evolution of individual menisci, and the growth of fingers within the porous microstructure. During that growth process, velocity jumps 20-25 times larger in magnitude than the bulk velocity were measured in the water phase and these bursts of water flow occurred both in-line with and against the bulk flow direction. These unsteady velocity events support the notion of pressure bursts and Haines jumps during pore drainage events as previously reported in the literature [1-3]. After passage of the CO2 front, shear-induced flow was detected in the trapped water ganglia in the form of circulation zones near the CO2-water interfaces as well as in the thin water films wetting the surfaces of the silicon micromodel. To our knowledge, the results presented herein represent the first quantitative spatially and temporally resolved velocity-field measurements at high pressure for water displacement by liquid/supercritical CO2 injection in a porous micromodel.

  12. A novel phosphoprotein analysis scheme for assessing changes in premalignant and malignant breast cell lines using 2D liquid separations, protein microarrays and tandem mass spectrometry

    PubMed Central

    Patwa, Tasneem H.; Wang, Yanfei; Miller, Fred R.; Goodison, Steve; Pennathur, Subramaniam; Barder, Timothy J.; Lubman, David M.

    2008-01-01

    An analysis of phosphorylation changes that occur during cancer progression would provide insights into the molecular pathways responsible for a malignant phenotype. In this study we employed a novel coupling of 2D-liquid separations and protein microarray technology to reveal changes in phosphoprotein status between premalignant (AT1) and malignant (CA1a) cell lines derived from the human MCF10A breast cell lines. Intact proteins were first separated according to their isoelectric point and hydrophobicities, then arrayed on SuperAmine glass slides. Phosphoproteins were detected using the universal, inorganic phospho-sensor dye, ProQ Diamond. Using this dye, out of 140 spots that were positive for phosphorylation, a total of 85 differentially expressed spots were detected over a pH range of 7.2 to 4.0. Proteins were identified and their peptides sequenced by mass spectrometry. The strategy enabled the identification of 75 differentially expressed phosphoproteins, from which 51 phosphorylation sites in 27 unique proteins were confirmed. Interestingly, the majority of differentially expressed phosphorylated proteins observed were nuclear proteins. Three regulators of apoptosis, Bad, Bax and Acinus, were also differentially phosphorylated in the two cell lines. Further development of this strategy will facilitate an understanding of the mechanisms involved in malignancy progression and other disease-related phenotypes. PMID:19194518

  13. Freestanding graphene paper decorated with 2D-assembly of Au@Pt nanoparticles as flexible biosensors to monitor live cell secretion of nitric oxide.

    PubMed

    Zan, Xiaoli; Fang, Zheng; Wu, Jin; Xiao, Fei; Huo, Fengwei; Duan, Hongwei

    2013-11-15

    We report the development of a new type of flexible electrochemical biosensors based on graphene paper loaded with closely-packed Au@Pt core-shell nanoparticles as a freestanding cell culture substrate for real-time monitoring cell secretion of nitric oxide. The hybrid electrode was fabricated through a modular approach in which 2D-assembly of nanoparticles formed at the oil-water interface was transferred onto graphene paper by dip-coating. We have shown that the independently optimized metal nanostructures and graphene paper were integrated into functional electrodes with high electrocatalytic activity. When used for the detection of nitric oxide, the flexible electrodes have demonstrated high sensitivity, a wide linear range, and a low detection limit, which, in combination with its biocompatibility, offer unique opportunities for the real-time monitoring of nitric oxide secretion by human endothelial vein cells grown on the electrode. These interesting findings collectively demonstrate the potential of our modular approach for designing high-performance flexible electrodes with tailored surface properties.

  14. Super pharmacological levels of calcitriol (1,25-(OH)2D3) inhibits mineral deposition and decreases cell proliferation in a strain dependent manner in chicken mesenchymal stem cells undergoing osteogenic differentiation in vitro

    PubMed Central

    Pande, Vivek V.; Chousalkar, Kapil C.; Bhanugopan, Marie S.; Quinn, Jane C.

    2015-01-01

    The biologically active form of vitamin D3, calcitriol (1,25-(OH)2D3), plays a key role in mineral homeostasis and bone formation and dietary vitamin D3 deficiency is a major cause of bone disorders in poultry. Supplementary dietary cholecalciferol (25-hydroxyvitamin D, 25-OH), the precursor of calcitriol, is commonly employed to combat this problem; however, dosage must be carefully determined as excess dietary vitamin D can cause toxicity resulting in a decrease in bone calcification, hypercalcinemia and renal failure. Despite much research on the therapeutic administration of dietary vitamin D in humans, the relative sensitivity of avian species to exogenous vitamin D has not been well defined. In order to determine the effects of exogenous 1,25-(OH)2D3 during avian osteogenesis, chicken bone marrow-derived mesenchymal stem cells (BM-MSCs) were exposed to varying doses of 1,25-(OH)2D3 during in vitro osteogenic differentiation and examined for markers of early proliferation and osteogenic induction. Similar to humans and other mammals, poultry BM-MSCs were found to be highly sensitive to exogenous 1,25-(OH)2D3 with super pharmacological levels exerting significant inhibition of mineralization and loss of cell proliferation in vitro. Strain related differences were apparent, with BM-MCSs derived from layers strains showing a higher level of sensitivity to 1,25-(OH)2D3 than those from broilers. These data suggest that understanding species and strain specific sensitivities to 1,25-(OH)2D3 is important for optimizing bone health in the poultry industry and that use of avian BM-MSCs are a useful tool for examining underlying effects of genetic variation in poultry. PMID:26500277

  15. Association between genetic taste sensitivity, 2D:4D ratio, dental caries prevalence, and salivary flow rate in 6-14-year-old children: a cross-sectional study

    PubMed Central

    Lakshmi, Chintamaneni Raja; Radhika, Doppalapudi; Prabhat, Mpv; Bhavana, Sujana mulk; Sai Madhavi, Nallamilli

    2016-01-01

    Background. The aim of this study was to assess the relationship between genetic taste sensitivity, dietary preferences and salivary flow rate in 6‒14-year-old children for identification of individuals at higher risk of developing dental caries. Methods. A total of 500 children 6‒14 years of age, of both genders, who reported to the Department of Oral Medicine and Radiology, were included. Propylthiouracil (PROP) sensitivity test was carried out and the subjects whose perception was bitter were grouped as tasters, whereas those who were unable to perceive any taste were grouped as non-tasters. The 2D:4D ratio was obtained by measuring the length ratio of index finger to ring finger with the help of a digital Vernier caliper. Evaluation of dietary preferences was carried out using a 24-hour dietary recall and accordingly they were categorized as sweet likers and dislikers. The salivary flow rate was estimated by collecting unstimulated saliva by spitting method. Data were analyzed with Student’s t-test and chi-squared test. Results. The results suggested a positive relation between low digit ratio (2D:4D), non-tasters, sweet likers and high caries index among the participants with a highly significant statistical difference (P ≤ 0.000). Tasters had high mean of USSR (0.48) than non-tasters (0.29), which was statistically significant. Conclusion. The present research revealed a positive correlation between all the parameters evaluated. Therefore an individual considered as non-taster by PROP was a sweet liker with low 2D:4D ratio, reduced salivary flow rate and high caries index.

  16. Association between genetic taste sensitivity, 2D:4D ratio, dental caries prevalence, and salivary flow rate in 6-14-year-old children: a cross-sectional study

    PubMed Central

    Lakshmi, Chintamaneni Raja; Radhika, Doppalapudi; Prabhat, Mpv; Bhavana, Sujana mulk; Sai Madhavi, Nallamilli

    2016-01-01

    Background. The aim of this study was to assess the relationship between genetic taste sensitivity, dietary preferences and salivary flow rate in 6‒14-year-old children for identification of individuals at higher risk of developing dental caries. Methods. A total of 500 children 6‒14 years of age, of both genders, who reported to the Department of Oral Medicine and Radiology, were included. Propylthiouracil (PROP) sensitivity test was carried out and the subjects whose perception was bitter were grouped as tasters, whereas those who were unable to perceive any taste were grouped as non-tasters. The 2D:4D ratio was obtained by measuring the length ratio of index finger to ring finger with the help of a digital Vernier caliper. Evaluation of dietary preferences was carried out using a 24-hour dietary recall and accordingly they were categorized as sweet likers and dislikers. The salivary flow rate was estimated by collecting unstimulated saliva by spitting method. Data were analyzed with Student’s t-test and chi-squared test. Results. The results suggested a positive relation between low digit ratio (2D:4D), non-tasters, sweet likers and high caries index among the participants with a highly significant statistical difference (P ≤ 0.000). Tasters had high mean of USSR (0.48) than non-tasters (0.29), which was statistically significant. Conclusion. The present research revealed a positive correlation between all the parameters evaluated. Therefore an individual considered as non-taster by PROP was a sweet liker with low 2D:4D ratio, reduced salivary flow rate and high caries index. PMID:27651879

  17. Association between genetic taste sensitivity, 2D:4D ratio, dental caries prevalence, and salivary flow rate in 6-14-year-old children: a cross-sectional study.

    PubMed

    Lakshmi, Chintamaneni Raja; Radhika, Doppalapudi; Prabhat, Mpv; Bhavana, Sujana Mulk; Sai Madhavi, Nallamilli

    2016-01-01

    Background. The aim of this study was to assess the relationship between genetic taste sensitivity, dietary preferences and salivary flow rate in 6‒14-year-old children for identification of individuals at higher risk of developing dental caries. Methods. A total of 500 children 6‒14 years of age, of both genders, who reported to the Department of Oral Medicine and Radiology, were included. Propylthiouracil (PROP) sensitivity test was carried out and the subjects whose perception was bitter were grouped as tasters, whereas those who were unable to perceive any taste were grouped as non-tasters. The 2D:4D ratio was obtained by measuring the length ratio of index finger to ring finger with the help of a digital Vernier caliper. Evaluation of dietary preferences was carried out using a 24-hour dietary recall and accordingly they were categorized as sweet likers and dislikers. The salivary flow rate was estimated by collecting unstimulated saliva by spitting method. Data were analyzed with Student's t-test and chi-squared test. Results. The results suggested a positive relation between low digit ratio (2D:4D), non-tasters, sweet likers and high caries index among the participants with a highly significant statistical difference (P ≤ 0.000). Tasters had high mean of USSR (0.48) than non-tasters (0.29), which was statistically significant. Conclusion. The present research revealed a positive correlation between all the parameters evaluated. Therefore an individual considered as non-taster by PROP was a sweet liker with low 2D:4D ratio, reduced salivary flow rate and high caries index. PMID:27651879

  18. Protection of rat intestinal epithelial cells from ischemia/reperfusion injury by (D-Ala2, D-Leu5)-enkephalin through inhibition of the MKK7-JNK signaling pathway

    PubMed Central

    WANG, ZHENRAN; TANG, BO; TANG, FANG; LI, YANG; ZHANG, GUANGYU; ZHONG, LI; DONG, CHENCHENG; HE, SONGQING

    2015-01-01

    Previous studies have demonstrated that (D-Ala2, D-Leu5)-enkephalin (DADLE) protects rats from hepatic ischemia/reperfusion (I/R) injury. In the present study, DADLE was also observed to alleviate IR-induced intestinal epithelial cell injury in rats by inhibiting mitogen-activated protein kinase kinase 7 (MKK7)-c-Jun N-terminal kinase (JNK) pathway signaling. To investigate the protective effect of DADLE on hypoxia/reoxygenation injury in rat intestinal epithelial cells, rat intestinal epithelial cells were treated with different concentrations of DADLE, following which the cell survival rate was determined using a tetrazolium (MTT) colorimetric assay, and apoptosis was determined using flow cytometry. To confirm whether the protective effect of DADLE was due to its effect on MKK7-JNK signaling, the phosphorylation levels of MKK7 and JNK were analyzed using western blot analysis following treatment with different concentrations of DADLE. The results demonstrated that, following treatment with DADLE, the survival rate of the rat intestinal cells subjected to I/R-induced injury increased significantly and the apoptotic rate decreased in a concentration-dependent manner. In addition, the levels of phosphorylated MKK7 and JNK decreased in a concentration-dependent manner following treatment with DADLE. Silencing the gene expression of MKK7 using small interfering RNA prior to DADLE treatment resulted in a reduction in the protective effects of DADLE on the rat intestinal epithelial cells subjected to I/R injury. Collectively, the results of the present study demonstrated that the protective effects of DADLE in I/R injury in rat intestinal cells occurred through inhibition of the MKK7-JNK pathway. PMID:26126577

  19. Non-Fullerene Polymer Solar Cells Based on Alkylthio and Fluorine Substituted 2D-Conjugated Polymers Reach 9.5% Efficiency.

    PubMed

    Bin, Haijun; Zhang, Zhi-Guo; Gao, Liang; Chen, Shanshan; Zhong, Lian; Xue, Lingwei; Yang, Changduk; Li, Yongfang

    2016-04-01

    Non-fullerene polymer solar cells (PSCs) with solution-processable n-type organic semiconductor (n-OS) as acceptor have seen rapid progress recently owing to the synthesis of new low bandgap n-OS, such as ITIC. To further increase power conversion efficiency (PCE) of the devices, it is of a great challenge to develop suitable polymer donor material that matches well with the low bandgap n-OS acceptors thus providing complementary absorption and nanoscaled blend morphology, as well as suppressed recombination and minimized energy loss. To address this challenge, we synthesized three medium bandgap 2D-conjugated bithienyl-benzodithiophene-alt-fluorobenzotriazole copolymers J52, J60, and J61 for the application as donor in the PSCs with low bandgap n-OS ITIC as acceptor. The three polymers were designed with branched alkyl (J52), branched alkylthio (J60), and linear alkylthio (J61) substituent on the thiophene conjugated side chain of the benzodithiophene (BDT) units for studying effect of the substituents on the photovoltaic performance of the polymers. The alkylthio side chain, red-shifted absorption down-shifted the highest occupied molecular orbital (HOMO) level and improved crystallinity of the 2D conjugated polymers. With linear alkylthio side chain, the tailored polymer J61 exhibits an enhanced JSC of 17.43 mA/cm(2), a high VOC of 0.89 V, and a PCE of 9.53% in the best non-fullerene PSCs with the polymer as donor and ITIC as acceptor. To the best of our knowledge, the PCE of 9.53% is one of the highest values reported in literature to date for the non-fullerene PSCs. The results indicate that J61 is a promising medium bandgap polymer donor in non-fullerene PSCs.

  20. Electrically rechargeable REDOX flow cell

    NASA Technical Reports Server (NTRS)

    Thaller, L. H. (Inventor)

    1976-01-01

    A bulk energy storage system is designed with an electrically rechargeable reduction-oxidation (REDOX) cell divided into two compartments by a membrane, each compartment containing an electrode. An anode fluid is directed through the first compartment at the same time that a cathode fluid is directed through the second compartment. Means are provided for circulating the anode and cathode fluids, and the electrodes are connected to an intermittent or non-continuous electrical source, which when operating, supplies current to a load as well as to the cell to recharge it. Ancillary circuitry is provided for disconnecting the intermittent source from the cell at prescribed times and for circulating the anode and cathode fluids according to desired parameters and conditions.

  1. Cellulose Structural Polymorphism in Plant Primary Cell Walls Investigated by High-Field 2D Solid-State NMR Spectroscopy and Density Functional Theory Calculations.

    PubMed

    Wang, Tuo; Yang, Hui; Kubicki, James D; Hong, Mei

    2016-06-13

    The native cellulose of bacterial, algal, and animal origins has been well studied structurally using X-ray and neutron diffraction and solid-state NMR spectroscopy, and is known to consist of varying proportions of two allomorphs, Iα and Iβ, which differ in hydrogen bonding, chain packing, and local conformation. In comparison, cellulose structure in plant primary cell walls is much less understood because plant cellulose has lower crystallinity and extensive interactions with matrix polysaccharides. Here we have combined two-dimensional magic-angle-spinning (MAS) solid-state nuclear magnetic resonance (solid-state NMR) spectroscopy at high magnetic fields with density functional theory (DFT) calculations to obtain detailed information about the structural polymorphism and spatial distributions of plant primary-wall cellulose. 2D (13)C-(13)C correlation spectra of uniformly (13)C-labeled cell walls of several model plants resolved seven sets of cellulose chemical shifts. Among these, five sets (denoted a-e) belong to cellulose in the interior of the microfibril while two sets (f and g) can be assigned to surface cellulose. Importantly, most of the interior cellulose (13)C chemical shifts differ significantly from the (13)C chemical shifts of the Iα and Iβ allomorphs, indicating that plant primary-wall cellulose has different conformations, packing, and hydrogen bonding from celluloses of other organisms. 2D (13)C-(13)C correlation experiments with long mixing times and with water polarization transfer revealed the spatial distributions and matrix-polysaccharide interactions of these cellulose structures. Celluloses f and g are well mixed chains on the microfibril surface, celluloses a and b are interior chains that are in molecular contact with the surface chains, while cellulose c resides in the core of the microfibril, outside spin diffusion contact with the surface. Interestingly, cellulose d, whose chemical shifts differ most significantly from those of

  2. Cellulose Structural Polymorphism in Plant Primary Cell Walls Investigated by High-Field 2D Solid-State NMR Spectroscopy and Density Functional Theory Calculations.

    PubMed

    Wang, Tuo; Yang, Hui; Kubicki, James D; Hong, Mei

    2016-06-13

    The native cellulose of bacterial, algal, and animal origins has been well studied structurally using X-ray and neutron diffraction and solid-state NMR spectroscopy, and is known to consist of varying proportions of two allomorphs, Iα and Iβ, which differ in hydrogen bonding, chain packing, and local conformation. In comparison, cellulose structure in plant primary cell walls is much less understood because plant cellulose has lower crystallinity and extensive interactions with matrix polysaccharides. Here we have combined two-dimensional magic-angle-spinning (MAS) solid-state nuclear magnetic resonance (solid-state NMR) spectroscopy at high magnetic fields with density functional theory (DFT) calculations to obtain detailed information about the structural polymorphism and spatial distributions of plant primary-wall cellulose. 2D (13)C-(13)C correlation spectra of uniformly (13)C-labeled cell walls of several model plants resolved seven sets of cellulose chemical shifts. Among these, five sets (denoted a-e) belong to cellulose in the interior of the microfibril while two sets (f and g) can be assigned to surface cellulose. Importantly, most of the interior cellulose (13)C chemical shifts differ significantly from the (13)C chemical shifts of the Iα and Iβ allomorphs, indicating that plant primary-wall cellulose has different conformations, packing, and hydrogen bonding from celluloses of other organisms. 2D (13)C-(13)C correlation experiments with long mixing times and with water polarization transfer revealed the spatial distributions and matrix-polysaccharide interactions of these cellulose structures. Celluloses f and g are well mixed chains on the microfibril surface, celluloses a and b are interior chains that are in molecular contact with the surface chains, while cellulose c resides in the core of the microfibril, outside spin diffusion contact with the surface. Interestingly, cellulose d, whose chemical shifts differ most significantly from those of

  3. Developing a flow control strategy to reduce nutrient load in a reclaimed multi-reservoir system using a 2D hydrodynamic and water quality model.

    PubMed

    Park, Yongeun; Cho, Kyung Hwa; Kang, Joo-Hyon; Lee, Seung Won; Kim, Joon Ha

    2014-01-01

    Blocking the natural bi-directional flow in an estuarine system using an artificial dyke has commonly caused serious water quality problems. In the southwestern part of South Korea, a parallel triple-reservoir system was constructed by blocking the mouth of three different rivers (Yeongsan, Okcheon, and Kumja), which were then interconnected using two open channels. This system has experienced a deterioration in water quality due to pollutants accumulated from the upper watershed, and has continually discharged pollutant loads to the outer ocean. Therefore, the objective of this study is to establish an effective dam operation plan for reducing nutrient loads released from the integrated reservoir. In this study, the CE-QUAL-W2 model, which is a 2-dimentional hydrodynamic and water quality model, was applied to predict the pollutant load released from each reservoir in response to different flow scenarios for the interconnecting channel. The model was calibrated using two novel methods: a sensitivity analysis to determine meaningful model parameters, and a pattern search to optimize the parameters. From the scenario analysis using flow control, it was determined that the total nitrogen (TN) and total phosphorus (TP) loadings could be reduced by 27.2% and 6.6%, respectively, under the optimal channel flow scenario by regulating the chlorophyll-a concentration in the reservoir. The results confirm that effective dam operation could contribute to a decrease in pollutant loads in the receiving seawater body. As such, this study suggests operational strategies for a multi-reservoir system that can be used to reduce the nutrient load being discharged from reservoirs.

  4. Pitot survey of exhaust flow field of a 2-D scramjet nozzle at Mach 6 with air or freon and argon used for exhaust simulation

    NASA Technical Reports Server (NTRS)

    Monta, William J.

    1992-01-01

    A pitot-rake survey of the simulated exhaust of a half-span scramjet nozzle model was conducted in the Langley 20-Inch Mach 6 Tunnel to provide an additional data set for computational fluid dynamics (CFD) code comparisons. A wind-tunnel model was tested with a 26-tube pitot rake that could be manually positioned along the mid-semispan plane of the model. The model configuration had an external expansion surface of 20 degrees and an internal cowl expansion of 12 degrees; tests were also performed with a flow fence. Tests were conducted at a free-stream Reynolds number of approximately 6.5 x 10(exp 6) per foot and a model angle of attack of -0.75 degrees. The two exhaust gas mediums that were tested were air and a Freon 12-argon mixture. Each medium was tested at two jet total pressures at approximately 28 and 14 psia. This document presents the flow-field survey results in graphical as well as tabular form, and several observations concerning the results are discussed. The surveys reveal the major expected flow-field characteristics for each test configuration. For a 50-percent freon 12 and 50-percent argon mixture by volume (Fr-Ar), the exhaust jet pressures were slightly higher than those for air. The addition of a flow fence slightly raised the pitot pressure for the Fr-Ar mixture, but it produced little change for air. For the Fr-Ar exhaust, the plume was larger and the region between the shock wave and plume was smaller.

  5. A numerical study of steady 2D flow around NACA 0015 and NACA 0012 hydrofoil with free surface using VOF method

    NASA Astrophysics Data System (ADS)

    Adjali, Saadia; Belkadi, Mustapha; Aounallah, Mohammed; Imine, Omar

    2015-05-01

    Accurate simulation of turbulent free surface flows around surface ships has a central role in the optimal design of such naval vessels. The flow problem to be simulated is rich in complexity and poses many modeling challenges because of the existence of breaking waves around the ship hull, and because of the interaction of the two-phase flow with the turbulent boundary layer. In this paper, our goal is to estimate the lift and drag coefficients for NACA 0012 of hydrofoil advancing in calm water under steady conditions with free surface and emerged NACA 0015. The commercial CFD software FLUENT version 14 is used for the computations in the present study. The calculated grid is established using the code computer GAMBIT 2.3.26.The shear stress k-ωSST model is used for turbulence modeling and the volume of fluid technique is employed to simulate the free-surface motion. In this computation, the second order upwind scheme is used for discretizing the convection terms in the momentum transport equations, the Modified HRIC scheme for VOF discretisation. The results obtained compare well with the experimental data.

  6. Electrochemical and impedance characterization of Microbial Fuel Cells based on 2D and 3D anodic electrodes working with seawater microorganisms under continuous operation.

    PubMed

    Hidalgo, D; Sacco, A; Hernández, S; Tommasi, T

    2015-11-01

    A mixed microbial population naturally presents in seawater was used as active anodic biofilm of two Microbial Fuel Cells (MFCs), employing either a 2D commercial carbon felt or 3D carbon-coated Berl saddles as anode electrodes, with the aim to compare their electrochemical behavior under continuous operation. After an initial increase of the maximum power density, the felt-based cell reduced its performance at 5 months (from 7 to 4 μW cm(-2)), while the saddle-based MFC exceeds 9 μW cm(-2) (after 2 months) and maintained such performance for all the tests. Electrochemical impedance spectroscopy was used to identify the MFCs controlling losses and indicates that the mass-transport limitations at the biofilm-electrolyte interface have the main contribution (>95%) to their internal resistance. The activation resistance was one order of magnitude lower with the Berl saddles than with carbon felt, suggesting an enhanced charge-transfer in the high surface-area 3D electrode, due to an increase in bacteria population growth.

  7. Expression of transcription factors after short-term exposure of Arabidopsis thaliana cell cultures to hypergravity and simulated microgravity (2-D/3-D clinorotation, magnetic levitation)

    NASA Astrophysics Data System (ADS)

    Babbick, M.; Dijkstra, C.; Larkin, O. J.; Anthony, P.; Davey, M. R.; Power, J. B.; Lowe, K. C.; Cogoli-Greuter, M.; Hampp, R.

    Gravity is an important environmental factor that controls plant growth and development. Studies have shown that the perception of gravity is not only a property of specialized cells, but can also be performed by undifferentiated cultured cells. In this investigation, callus of Arabidopsis thaliana cv. Columbia was used to investigate the initial steps of gravity-related signalling cascades, through altered expression of transcription factors (TFs). TFs are families of small proteins that regulate gene expression by binding to specific promoter sequences. Based on microarray studies, members of the gene families WRKY, MADS-box, MYB, and AP2/EREBP were selected for investigation, as well as members of signalling chains, namely IAA 19 and phosphoinositol-4-kinase. Using qRT-PCR, transcripts were quantified within a period of 30 min in response to hypergravity (8 g), clinorotation [2-D clinostat and 3-D random positioning machine (RPM)] and magnetic levitation (ML). The data indicated that (1) changes in gravity induced stress-related signalling, and (2) exposure in the RPM induced changes in gene expression which resemble those of magnetic levitation. Two dimensional clinorotation resulted in responses similar to those caused by hypergravity. It is suggested that RPM and ML are preferable to simulate microgravity than clinorotation.

  8. Electrochemical and impedance characterization of Microbial Fuel Cells based on 2D and 3D anodic electrodes working with seawater microorganisms under continuous operation.

    PubMed

    Hidalgo, D; Sacco, A; Hernández, S; Tommasi, T

    2015-11-01

    A mixed microbial population naturally presents in seawater was used as active anodic biofilm of two Microbial Fuel Cells (MFCs), employing either a 2D commercial carbon felt or 3D carbon-coated Berl saddles as anode electrodes, with the aim to compare their electrochemical behavior under continuous operation. After an initial increase of the maximum power density, the felt-based cell reduced its performance at 5 months (from 7 to 4 μW cm(-2)), while the saddle-based MFC exceeds 9 μW cm(-2) (after 2 months) and maintained such performance for all the tests. Electrochemical impedance spectroscopy was used to identify the MFCs controlling losses and indicates that the mass-transport limitations at the biofilm-electrolyte interface have the main contribution (>95%) to their internal resistance. The activation resistance was one order of magnitude lower with the Berl saddles than with carbon felt, suggesting an enhanced charge-transfer in the high surface-area 3D electrode, due to an increase in bacteria population growth. PMID:26166463

  9. Geophysical Fluid Flow Cell (GFFC) Simulation

    NASA Technical Reports Server (NTRS)

    1999-01-01

    These simulations of atmospheric flow use the same experimental parameters but started with slightly different initial conditions in the model. The simulations were part of data analysis for the Geophysical Fluid Flow Cell (GFFC), a planet in a test tube apparatus flown on Spacelab to mimic the atmospheres on gas giant planets and stars. (Credit: Dr. Tim Miller of Global Hydrology and Climate Center at the Marshall Space Flight Center)

  10. Poly(amido amine)-based multilayered thin films on 2D and 3D supports for surface-mediated cell transfection.

    PubMed

    Hujaya, Sry D; Marchioli, Giulia; Roelofs, Karin; van Apeldoorn, Aart A; Moroni, Lorenzo; Karperien, Marcel; Paulusse, Jos M J; Engbersen, Johan F J

    2015-05-10

    Two linear poly(amido amine)s, pCABOL and pCHIS, prepared by polyaddition of cystamine bisacrylamide (C) with 4-aminobutanol (ABOL) or histamine (HIS), were explored to form alternating multilayer thin films with DNA to obtain functionalized materials with transfection capacity in 2D and 3D. Therefore, COS-7 cells were cultured on top of multilayer films formed by layer-by-layer dipcoating of these polymers with GFP-encoded pDNA, and the effect of the number of layers and cell seeding density on the transfection efficiency was evaluated. Multilayer films with pCABOL were found to be superior to pCHIS in facilitating transfection, which was attributed to higher incorporation of pDNA and release of the transfection agent. High amounts of transfected cells were obtained on pCABOL films, correlating proportionally over a wide range with seeding density. Optimal transfection efficiency was obtained with pCABOL films composed of 10 bilayers. Further increase in the number of bilayers only marginally increased transfection efficiency. Using the optimal multilayer and cell seeding conditions, pCABOL multilayers were fabricated on poly(ε-caprolactone) (PCL), heparinized PCL (PCL-HEP), and poly(lactic acid) (PLA) disks as examples of common biomedical supports. The multilayers were found to completely mask the properties of the original substrates, with significant improvement in cell adhesion, which is especially pronounced for PCL and PLA disks. With all these substrates, transfection efficiency was found to be in the range of 25-50% transfected cells. The pCABOL/pDNA multilayer films can also conveniently add transfection capability to 3D scaffolds. Significant improvement in cell adhesion was observed after multilayer coating of 3D-plotted fibers of PCL (with and without an additional covalent heparin layer), especially for the PCL scaffold without heparin layer and transfection was observed on both 3D PCL and PCL-HEP scaffolds. These results show that layer

  11. Transfer of the human NKG2D ligands UL16 binding proteins (ULBP) 1-3 is related to lytic granule release and leads to ligand retransfer and killing of ULBP-recipient natural killer cells.

    PubMed

    López-Cobo, Sheila; Romera-Cárdenas, Gema; García-Cuesta, Eva M; Reyburn, Hugh T; Valés-Gómez, Mar

    2015-09-01

    After immune interactions, membrane fragments can be transferred between cells. This fast transfer of molecules is transient and shows selectivity for certain proteins; however, the constraints underlying acquisition of a protein are unknown. To characterize the mechanism and functional consequences of this process in natural killer (NK) cells, we have compared the transfer of different NKG2D ligands. We show that human NKG2D ligands can be acquired by NK cells with different efficiencies. The main findings are that NKG2D ligand transfer is related to immune activation and receptor-ligand interaction and that NK cells acquire these proteins during interactions with target cells that lead to degranulation. Our results further demonstrate that NK cells that have acquired NKG2D ligands can stimulate activation of autologous NK cells. Surprisingly, NK cells can also re-transfer the acquired molecule to autologous effector cells during this immune recognition that leads to their death. These data demonstrate that transfer of molecules occurs as a consequence of immune recognition and imply that this process might play a role in homeostatic tuning-down of the immune response or be used as marker of interaction.

  12. Flow over glycocalyx covered endothelial cells.

    NASA Astrophysics Data System (ADS)

    Waters, Sarah L.; Liu, Shu Q.; Grotberg, James B.

    1997-11-01

    Blood vessels are lined with a monolayer of endothelial cells that interact with the blood flow. Two structural features of the endothelial membrane may influence the fluid dynamics over the endothelium: 1) the endothelial membrane bumpiness; and 2) the porous glycocalyx layer covering the cell surface. The bumpiness of the endothelium may induce regions of recirculation, and the glycocalyx may effect the flow pattern at the cell surface. An analytical study is presented for pressure driven blood flow in the microcirculation. The vessel is modeled as a rigid, impermeable, symmetric two--dimensional channel, which has sinusoidally wavy walls. The vessel has two regions: 1) the glycocalyx layer which is modeled as a uniformly thick poroelastic deformable wall layer using biphasic mixture theory; and 2) the free lumen where the Navier--Stokes equations of motion apply. Analytical results are obtained by making the long wavelength approximation. The model predicts the fluid flow and hence the shear stress exerted by the flow on the individual endothelial cells and at the glycocalyx--lumen interface. Implications of the results for biological events such as molecular transport and signal transduction are considered.

  13. Comparison of 2D and 3D Computational Multiphase Fluid Flow Models of Oxygen Lancing of Pyrometallurgical Furnace Tap-Holes

    NASA Astrophysics Data System (ADS)

    Erwee, M. W.; Reynolds, Q. G.; Zietsman, J. H.

    2016-06-01

    Furnace tap-holes vary in design depending on the type of furnace and process involved, but they share one common trait: The tap-hole must be opened and closed periodically. In general, tap-holes are plugged with refractory clay after tapping, thereby stopping the flow of molten material. Once a furnace is ready to be tapped, drilling and/or lancing with oxygen are typically used to remove tap-hole clay from the tap-hole. Lancing with oxygen is an energy-intensive, mostly manual process, which affects the performance and longevity of the tap-hole refractory material as well as the processes inside the furnace. Computational modeling offers an opportunity to gain insight into the possible effects of oxygen lancing on various aspects of furnace operation.

  14. Free flow cell electrophoresis using zwitterionic buffer

    NASA Technical Reports Server (NTRS)

    Rodkey, R. Scott

    1990-01-01

    Studies of a zwitterionic buffer formulated for cell electrophoresis were done using the McDonnell-Douglas Continuous Flow Electrophoresis System. Standard buffers were analyzed for their stability in the electrical field and the results showed that both buffers tested were inherently unstable. Further, titration studies showed that the standards buffers buffered poorly at the pH employed for electrophoresis. The zwitterionic buffer buffered well at its nominal pH and was shown to be stable in the electrical field. Comparative studies of the buffer with standard cell separation buffers using formalin fixed rabbit and goose red blood cells showed that the zwitterionic buffer gave better resolution of the fixed cells. Studies with viable hybridoma cells showed that buffer Q supported cell viability equal to Hank's Balanced Salt Solution and that hybridoma cells in different stages of the growth cycle demonstrated reproducible differences in electrophoretic mobility.

  15. Comparative 2D-DIGE Proteomic Analysis of Bovine Mammary Epithelial Cells during Lactation Reveals Protein Signatures for Lactation Persistency and Milk Yield

    PubMed Central

    Janjanam, Jagadeesh; Singh, Surender; Jena, Manoj K.; Varshney, Nishant; Kola, Srujana; Kumar, Sudarshan; Kaushik, Jai K.; Grover, Sunita; Dang, Ajay K.; Mukesh, Manishi; Prakash, B. S.; Mohanty, Ashok K.

    2014-01-01

    Mammary gland is made up of a branching network of ducts that end with alveoli which surrounds the lumen. These alveolar mammary epithelial cells (MEC) reflect the milk producing ability of farm animals. In this study, we have used 2D-DIGE and mass spectrometry to identify the protein changes in MEC during immediate early, peak and late stages of lactation and also compared differentially expressed proteins in MEC isolated from milk of high and low milk producing cows. We have identified 41 differentially expressed proteins during lactation stages and 22 proteins in high and low milk yielding cows. Bioinformatics analysis showed that a majority of the differentially expressed proteins are associated in metabolic process, catalytic and binding activity. The differentially expressed proteins were mapped to the available biological pathways and networks involved in lactation. The proteins up-regulated during late stage of lactation are associated with NF-κB stress induced signaling pathways and whereas Akt, PI3K and p38/MAPK signaling pathways are associated with high milk production mediated through insulin hormone signaling. PMID:25111801

  16. Cell migration does not produce membrane flow

    PubMed Central

    1990-01-01

    We have previously reported that rearward migration of surface particles on slowly moving cells is not driven by membrane flow (Sheetz, M. P., S. Turney, H. Qian, and E. L. Elson. 1989. Nature (Lond.). 340:284-288) and recent photobleaching measurements have ruled out any rapid rearward lipid flow (Lee, J., M. Gustafsson, D. E. Magnussen, and K. Jacobson. 1990. Science (Wash. DC.) 247:1229-1233). It was not possible, however, to conclude from those studies that a slower or tank-tread membrane lipid flow does not occur. Therefore, we have used the technology of single particle tracking to examine the movements of diffusing particles on rapidly locomoting fish keratocytes where the membrane current is likely to be greatest. The keratocytes had a smooth lamellipodial surface on which bound Con A-coated gold particles were observed either to track toward the nuclear region (velocity of 0.35 +/- 0.15 micron/s) or to diffuse randomly (apparent diffusion coefficient of [3.5 +/- 2.0] x 10(-10) cm2/s). We detected no systematic drift relative to the cell edge of particles undergoing random diffusion even after the cell had moved many micrometers. The average net particle displacement was 0.01 +/- 2.7% of the cell displacement. These results strongly suggest that neither the motions of membrane proteins driven by the cytoskeleton nor other possible factors produce a bulk flow of membrane lipid. PMID:2211827

  17. Time-lapse microscopy and classification of 2D human mesenchymal stem cells based on cell shape picks up myogenic from osteogenic and adipogenic differentiation.

    PubMed

    Seiler, Christof; Gazdhar, Amiq; Reyes, Mauricio; Benneker, Lorin M; Geiser, Thomas; Siebenrock, Klaus A; Gantenbein-Ritter, Benjamin

    2014-09-01

    Current methods to characterize mesenchymal stem cells (MSCs) are limited to CD marker expression, plastic adherence and their ability to differentiate into adipogenic, osteogenic and chondrogenic precursors. It seems evident that stem cells undergoing differentiation should differ in many aspects, such as morphology and possibly also behaviour; however, such a correlation has not yet been exploited for fate prediction of MSCs. Primary human MSCs from bone marrow were expanded and pelleted to form high-density cultures and were then randomly divided into four groups to differentiate into adipogenic, osteogenic chondrogenic and myogenic progenitor cells. The cells were expanded as heterogeneous and tracked with time-lapse microscopy to record cell shape, using phase-contrast microscopy. The cells were segmented using a custom-made image-processing pipeline. Seven morphological features were extracted for each of the segmented cells. Statistical analysis was performed on the seven-dimensional feature vectors, using a tree-like classification method. Differentiation of cells was monitored with key marker genes and histology. Cells in differentiation media were expressing the key genes for each of the three pathways after 21 days, i.e. adipogenic, osteogenic and chondrogenic, which was also confirmed by histological staining. Time-lapse microscopy data were obtained and contained new evidence that two cell shape features, eccentricity and filopodia (= 'fingers') are highly informative to classify myogenic differentiation from all others. However, no robust classifiers could be identified for the other cell differentiation paths. The results suggest that non-invasive automated time-lapse microscopy could potentially be used to predict the stem cell fate of hMSCs for clinical application, based on morphology for earlier time-points. The classification is challenged by cell density, proliferation and possible unknown donor-specific factors, which affect the performance of

  18. Tracking Epithelial Cell Junctions in C. elegans Embryogenesis With Active Contours Guided by SIFT Flow

    PubMed Central

    Lee, Chen-Yu; Gonçalves, Monira; Chisholm, Andrew D.; Cosman, Pamela C.

    2015-01-01

    Quantitative analysis of cell shape in live samples is an important goal in developmental biology. Automated or semiautomated segmentation and tracking of cell nuclei has been successfully implemented in several biological systems. Segmentation and tracking of cell surfaces has been more challenging. Here, we present a new approach to tracking cell junctions in the developing epidermis of C. elegans embryos. Epithelial junctions as visualized with DLG-1::GFP form lines at the subapical circumference of differentiated epidermal cells and delineate changes in epidermal cell shape and position. We develop and compare two approaches for junction segmentation. For the first method (projection approach), 3-D cell boundaries are projected into 2D for segmentation using active contours with a nonintersecting force, and subsequently tracked using scale-invariant feature transform (SIFT) flow. The resulting 2-D tracked boundaries are then back-projected into 3-D space. The second method (volumetric approach) uses a 3-D extended version of active contours guided by SIFT flow in 3-D space. In both methods, cell junctions are manually located at the first time point and tracked in a fully automated way for the remainder of the video. Using these methods, we have generated the first quantitative description of ventral epidermal cell movements and shape changes during epidermal enclosure. PMID:24771564

  19. Artificial Hair Cells for Sensing Flows

    NASA Technical Reports Server (NTRS)

    Chen, Jack

    2007-01-01

    The purpose of this article is to present additional information about the flow-velocity sensors described briefly in the immediately preceding article. As noted therein, these sensors can be characterized as artificial hair cells that implement an approximation of the sensory principle of flow-sensing cilia of fish: A cilium is bent by an amount proportional to the flow to which it is exposed. A nerve cell at the base of the cilium senses the flow by sensing the bending of the cilium. In an artificial hair cell, the artificial cilium is a microscopic cantilever beam, and the bending of an artificial cilium is measured by means of a strain gauge at its base (see Figure 1). Figure 2 presents cross sections of a representative sensor of this type at two different stages of its fabrication process. The process consists of relatively- low-temperature metallization, polymer-deposition, microfabrication, and surface-micromachining subprocesses, including plastic-deformation magnetic assembly (PDMA), which is described below. These subprocesses are suitable for a variety of substrate materials, including silicon, some glasses, and some polymers. Moreover, because it incorporates a polymeric supporting structure, this sensor is more robust, relative to its silicon-based counterparts.

  20. 2D immunoblots show differential response of mouse IgG and IgM antibodies to antigens of mammary carcinoma 4 T1 cells

    PubMed Central

    2014-01-01

    Background Immunosuppression in breast cancer has been reported in women and in the highly metastatic mouse mammary tumor model 4 T1. The immunosuppressive environment complicates the use of the humoral response against the tumor as an immunodiagnostic tool. IgM has not been used in immunodiagnostic in part because its antitumor responses, both innate and adaptive, have not been studied in function of time in breast cancer. We show a new approach to analyzing the mouse humoral immune response, and compare the evolution with time of IgG and IgM responses against the antigens of 4 T1 cells. Methods The study is based on 2-dimensional immunoblotting detection of antigens from 4 T1 cells by the IgG and IgM antibodies in the serum of female mice injected with 4 T1 cells. Results There was a high variability in the intra-and inter-mouse response. Variability in the IgM response was manifested as a pattern of spots that could become a multibinomial variable of 0 and 1, which could represent a signature of the immune response. Different numbers of spots was found in the IgG and IgM responses from week 1 to 5. On average, the IgM had more but the IgG response decrease with the time. The natural IgM at t = 0 responds stronger than w1; the adaptive response of both IgM and IgG were elicited where, with the former being stronger better than the latter. Antigens that are recognized by some female mice in the first week are also recognized by other female mice at time 0. Contamination of the natural IgM makes difficult use the adaptive IgM as a tool for immunodiagnostic. Conclusions IgM and IgG response varied with the time and individuals. Spot variation in 2D pattern for the natural IgM could be expressed as a binomial signature, which opens up the way to correlate a particular pattern with resistance or susceptibility. This uncovers a battery of IgMs for each individual to confront cancer or infections. The possibility to differentiate between adaptive IgM antibodies

  1. Biofilm responses to smooth flow fields and chemical gradients in novel microfluidic flow cells

    PubMed Central

    Song, Jisun L.; Au, Kelly H.; Huynh, Kimberly T.

    2013-01-01

    We present two novel microfluidic flow cells developed to provide reliable control of flow distributions and chemical gradients in biofilm studies. We developed a single-inlet microfluidic flow cell to support biofilm growth under a uniform velocity field, and a double-inlet flow cell to provide a very smooth transverse concentration gradient. Both flow cells consist of a layer of polydimethylsiloxane (PDMS) bonded to glass cover slips and were fabricated using the replica molding technique. We demonstrate the capabilities of the flow cells by quantifying flow patterns before and after growth of Pseudomonas aeruginosa biofilms through particle imaging velocimetry, and by evaluating concentration gradients within the double-inlet microfluidic flow cell. Biofilm growth substantially increased flow complexity by diverting flow around biomass, creating high- and low-velocity regions and surface friction. Under a glucose gradient in the double-inlet flow cell, P. aeruginosa biofilms grew in proportion to the local glucose concentration, producing distinct spatial patterns in biofilm biomass relative to the imposed glucose gradient. When biofilms were subjected to a ciprofloxacin gradient, spatial patterns of fractions of dead cells were also in proportion to the local antibiotic concentration. These results demonstrate that the microfluidic flow cells are suitable for quantifying flow complexities resulting from flow-biofilm interactions and investigating spatial patterns of biofilm growth under chemical gradients. These novel microfluidic flow cells will facilitate biofilm research that requires flow control and in situ imaging, particularly investigations of biofilm-environment interactions. PMID:24038055

  2. Biofilm responses to smooth flow fields and chemical gradients in novel microfluidic flow cells.

    PubMed

    Song, Jisun L; Au, Kelly H; Huynh, Kimberly T; Packman, Aaron I

    2014-03-01

    We present two novel microfluidic flow cells developed to provide reliable control of flow distributions and chemical gradients in biofilm studies. We developed a single-inlet microfluidic flow cell to support biofilm growth under a uniform velocity field, and a double-inlet flow cell to provide a very smooth transverse concentration gradient. Both flow cells consist of a layer of polydimethylsiloxane (PDMS) bonded to glass cover slips and were fabricated using the replica molding technique. We demonstrate the capabilities of the flow cells by quantifying flow patterns before and after growth of Pseudomonas aeruginosa biofilms through particle imaging velocimetry, and by evaluating concentration gradients within the double-inlet microfluidic flow cell. Biofilm growth substantially increased flow complexity by diverting flow around biomass, creating high- and low-velocity regions and surface friction. Under a glucose gradient in the double-inlet flow cell, P. aeruginosa biofilms grew in proportion to the local glucose concentration, producing distinct spatial patterns in biofilm biomass relative to the imposed glucose gradient. When biofilms were subjected to a ciprofloxacin gradient, spatial patterns of fractions of dead cells were also in proportion to the local antibiotic concentration. These results demonstrate that the microfluidic flow cells are suitable for quantifying flow complexities resulting from flow-biofilm interactions and investigating spatial patterns of biofilm growth under chemical gradients. These novel microfluidic flow cells will facilitate biofilm research that requires flow control and in situ imaging, particularly investigations of biofilm-environment interactions. PMID:24038055

  3. High speed flow cytometric separation of viable cells

    DOEpatents

    Sasaki, D.T.; Van den Engh, G.J.; Buckie, A.M.

    1995-11-14

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  4. High speed flow cytometric separation of viable cells

    DOEpatents

    Sasaki, Dennis T.; Van den Engh, Gerrit J.; Buckie, Anne-Marie

    1995-01-01

    Hematopoietic cell populations are separated to provide cell sets and subsets as viable cells with high purity and high yields, based on the number of original cells present in the mixture. High-speed flow cytometry is employed using light characteristics of the cells to separate the cells, where high flow speeds are used to reduce the sorting time.

  5. Cytotoxicity of CD56(bright) NK cells towards autologous activated CD4+ T cells is mediated through NKG2D, LFA-1 and TRAIL and dampened via CD94/NKG2A.

    PubMed

    Nielsen, Natasja; Ødum, Niels; Ursø, Birgitte; Lanier, Lewis L; Spee, Pieter

    2012-01-01

    In mouse models of chronic inflammatory diseases, Natural Killer (NK) cells can play an immunoregulatory role by eliminating chronically activated leukocytes. Indirect evidence suggests that NK cells may also be immunoregulatory in humans. Two subsets of human NK cells can be phenotypically distinguished as CD16(+)CD56(dim) and CD16(dim/-)CD56(bright). An expansion in the CD56(bright) NK cell subset has been associated with clinical responses to therapy in various autoimmune diseases, suggesting an immunoregulatory role for this subset in vivo. Here we compared the regulation of activated human CD4(+) T cells by CD56(dim) and CD56(bright) autologous NK cells in vitro. Both subsets efficiently killed activated, but not resting, CD4(+) T cells. The activating receptor NKG2D, as well as the integrin LFA-1 and the TRAIL pathway, played important roles in this process. Degranulation by NK cells towards activated CD4(+) T cells was enhanced by IL-2, IL-15, IL-12+IL-18 and IFN-α. Interestingly, IL-7 and IL-21 stimulated degranulation by CD56(bright) NK cells but not by CD56(dim) NK cells. NK cell killing of activated CD4(+) T cells was suppressed by HLA-E on CD4(+) T cells, as blocking the interaction between HLA-E and the inhibitory CD94/NKG2A NK cell receptor enhanced NK cell degranulation. This study provides new insight into CD56(dim) and CD56(bright) NK cell-mediated elimination of activated autologous CD4(+) T cells, which potentially may provide an opportunity for therapeutic treatment of chronic inflammation. PMID:22384114

  6. Cell stretching in extensional flows for assaying cell mechanics

    NASA Astrophysics Data System (ADS)

    Gossett, Daniel; Tse, Henry; Adeyiga, Oladunni; Yang, Otto; Rao, Jianyu; di Carlo, Dino

    2013-03-01

    There is growing evidence that cell deformability is a useful indicator of cell state and may be a label-free biomarker of metastatic potential, degree of differentiation, and leukocyte activation. In order for deformability measurements to be clinically valuable given the heterogeneity of biological samples, there exists a need for a high-throughput assay of this biophysical property. We developed a robust method for obtaining high-throughput (>1,000 cells/sec) single-cell mechanical measurements which employs coupled hydrodynamic lift forces and curvature-induced secondary flows to uniformly position cells in flow, extensional flow stretching, high-speed imaging, and automated image analysis to extract diameter and deformability parameters. Using this method we have assayed numerous in vitro models of cellular transformations and clinical fluids where malignant cells manifest. We found transformations associated with increased motility or invasiveness increased deformability and the presence of large and deformable cells within clinical pleural fluids correlated well with cytological diagnoses of malignancy. This agrees with the hypothesis that cancerous cells are deformable by necessity-to be able to transverse tight endothelial gaps and invade tissues.

  7. Deterministic Aperiodic Sickle Cell Blood Flows

    NASA Astrophysics Data System (ADS)

    Atsaves, Louis; Harris, Wesley

    2013-11-01

    In this paper sickle cell blood flow in the capillaries is modeled as a hydrodynamical system. The hydrodynamical system consists of the axisymmetric unsteady, incompressible Navier-Stokes equations and a set of constitutive equations for oxygen transport. Blood cell deformation is not considered in this paper. The hydrodynamical system is reduced to a system of non-linear partial differential equations that are then transformed into a system of three autonomous non-linear ordinary differential equations and a set of algebraic equations. We examine the hydrodynamical system to discern stable/unstable, periodic/nonperiodic, reversible/irreversible properties of the system. The properties of the solutions are driven in large part by the coefficients of the governing system of equations. These coefficients depend on the physiological properties of the sickle cell blood. The chaotic nature of the onset of crisis in sickle cell patients is identified. Research Assistant.

  8. A novel bispecific protein (ULBP2-BB4) targeting the NKG2D receptor on natural killer (NK) cells and CD138 activates NK cells and has potent antitumor activity against human multiple myeloma in vitro and in vivo.

    PubMed

    von Strandmann, Elke Pogge; Hansen, Hinrich P; Reiners, Katrin S; Schnell, Roland; Borchmann, Peter; Merkert, Sabine; Simhadri, Venkateswara R; Draube, Andreas; Reiser, Marcel; Purr, Ingvill; Hallek, Michael; Engert, Andreas

    2006-03-01

    The inability of the immune system to recognize and kill malignant plasma cells in patients with multiple myeloma (MM) has been attributed in part to the ineffective activation of natural killer (NK) cells. In order to activate and target NK cells to the malignant cells in MM we designed a novel recombinant bispecific protein (ULBP2-BB4). While ULBP2 binds the activating NK receptor NKG2D, the BB4 moiety binds to CD138, which is overexpressed on a variety of malignancies, including MM. ULBP2-BB4 strongly activated primary NK cells as demonstrated by a significant increase in interferon-gamma (IFN-gamma) secretion. In vitro, ULBP2-BB4 enhanced the NK-mediated lysis of 2 CD138+ human MM cell lines, U-266 and RPMI-8226, and of primary malignant plasma cells in the allogenic and autologous setting. Moreover, in a nude mouse model with subcutaneously growing RPMI-8226 cells, the cotherapy with ULBP-BB4 and human peripheral blood lymphocytes abrogated the tumor growth. These data suggest potential clinical use of this novel construct in patients with MM. The use of recombinant NK receptor ligands that target NK cells to tumor cells might offer new approaches for other malignancies provided a tumor antigen-specific antibody is available.

  9. High divergent 2D grating

    NASA Astrophysics Data System (ADS)

    Wang, Jin; Ma, Jianyong; Zhou, Changhe

    2014-11-01

    A 3×3 high divergent 2D-grating with period of 3.842μm at wavelength of 850nm under normal incidence is designed and fabricated in this paper. This high divergent 2D-grating is designed by the vector theory. The Rigorous Coupled Wave Analysis (RCWA) in association with the simulated annealing (SA) is adopted to calculate and optimize this 2D-grating.The properties of this grating are also investigated by the RCWA. The diffraction angles are more than 10 degrees in the whole wavelength band, which are bigger than the traditional 2D-grating. In addition, the small period of grating increases the difficulties of fabrication. So we fabricate the 2D-gratings by direct laser writing (DLW) instead of traditional manufacturing method. Then the method of ICP etching is used to obtain the high divergent 2D-grating.

  10. A low-dimensional deformation model for cancer cells in flow.

    PubMed

    Lee, A M; Berny-Lang, M A; Liao, S; Kanso, E; Kuhn, P; McCarty, O J T; Newton, P K

    2012-08-01

    A low-dimensional parametric deformation model of a cancer cell under shear flow is developed. The model is built around an experiment in which MDA-MB-231 adherent cells are subjected to flow with increasing shear. The cell surface deformation is imaged using differential interference contrast microscopy imaging techniques until the cell releases into the flow. We post-process the time sequence of images using an active shape model from which we obtain the principal components of deformation. These principal components are then used to obtain the parameters in an empirical constitutive equation determining the cell deformations as a function of the fluid normal and shear forces imparted. The cell surface is modeled as a 2D Gaussian interface which can be deformed with three active parameters: H (height), σ(x) (x-width), and σ(y) (y-width). Fluid forces are calculated on the cell surface by discretizing the surface with regularized Stokeslets, and the flow is driven by a stochastically fluctuating pressure gradient. The Stokeslet strengths are obtained so that viscous boundary conditions are enforced on the surface of the cell and the surrounding plate. We show that the low-dimensional model is able to capture the principal deformations of the cell reasonably well and argue that active shape models can be exploited further as a useful tool to bridge the gap between experiments, models, and numerical simulations in this biological setting.

  11. Red blood cell in simple shear flow

    NASA Astrophysics Data System (ADS)

    Chien, Wei; Hew, Yayu; Chen, Yeng-Long

    2013-03-01

    The dynamics of red blood cells (RBC) in blood flow is critical for oxygen transport, and it also influences inflammation (white blood cells), thrombosis (platelets), and circulatory tumor migration. The physical properties of a RBC can be captured by modeling RBC as lipid membrane linked to a cytoskeletal spectrin network that encapsulates cytoplasm rich in hemoglobin, with bi-concave equilibrium shape. Depending on the shear force, RBC elasticity, membrane viscosity, and cytoplasm viscosity, RBC can undergo tumbling, tank-treading, or oscillatory motion. We investigate the dynamic state diagram of RBC in shear and pressure-driven flow using a combined immersed boundary-lattice Boltzmann method with a multi-scale RBC model that accurately captures the experimentally established RBC force-deformation relation. It is found that the tumbling (TU) to tank-treading (TT) transition occurs as shear rate increases for cytoplasm/outer fluid viscosity ratio smaller than 0.67. The TU frequency is found to be half of the TT frequency, in agreement with experiment observations. Larger viscosity ratios lead to the disappearance of stable TT phase and unstable complex dynamics, including the oscillation of the symmetry axis of the bi-concave shape perpendicular to the flow direction. The dependence on RBC bending rigidity, shear modulus, the order of membrane spectrin network and fluid field in the unstable region will also be discussed.

  12. Predictive value of endometrial thickness, pattern and sub-endometrial blood flows on the day of hCG by 2D doppler in in-vitro fertilization cycles: A prospective clinical study from a tertiary care unit

    PubMed Central

    Singh, Neeta; Bahadur, Anupama; Mittal, Suneeta; Malhotra, Neena; Bhatt, Ashok

    2011-01-01

    AIMS AND OBJECTIVES: To evaluate the role of endometrial thickness, pattern and sub-endometrial blood flows measured by 2D power Doppler ultrasound to predict pregnancy during in-vitro fertilization (IVF) treatment. STUDY DESIGN: Prospective, non-randomized clinical study. MATERIALS AND METHODS: This was a prospective observational study. A total of 101 infertile women were recruited from our IVF-ET program from January to December, 2009. Women with tubal factor, male factor and unexplained infertility were included in the study. RESULTS: The mean age was 35 years and mean duration of infertility was 8 years. Seventy five (74.25%) patients had primary infertility and 26 (25.74%) had secondary infertility. The mean endometrial thickness was 8.1 mm and endometrial blood flow was in Zone I in 18 patients, 28 patients had blood flow in Zone II and 54 had in Zone III. Overall, 27 (26.73%) patients conceived and in these women the endometrial thickness was between 6 and 12 mm. CONCLUSIONS: With a thin endometrium (≤7 mm) and no-triple-line endometrial pattern coexisting in an in-vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) candidate, cryopreservation should be recommended. With a thin endometrium and a good texture (triple-line), other prognostic factors, such as embryo quality, should be taken into account. The endometrial vascularity has a useful predictive value on the implantation rate in IVF cycles irrespective of the morphological appearance of the endometrium. However, further study is needed to make a definitive conclusion. PMID:21772737

  13. The Geophysical Fluid Flow Cell Experiment

    NASA Technical Reports Server (NTRS)

    Hart, J. E.; Ohlsen, D.; Kittleman, S.; Borhani, N.; Leslie, F.; Miller, T.

    1999-01-01

    The Geophysical Fluid Flow Cell (GFFC) experiment performed visualizations of thermal convection in a rotating differentially heated spherical shell of fluid. In these experiments dielectric polarization forces are used to generate a radially directed buoyancy force. This enables the laboratory simulation of a number of geophysically and astrophysically important situations in which sphericity and rotation both impose strong constraints on global scale fluid motions. During USML-2 a large set of experiments with spherically symmetric heating were carried out. These enabled the determination of critical points for the transition to various forms of nonaxisymmetric convection and, for highly turbulent flows, the transition latitudes separating the different modes of motion. This paper presents a first analysis of these experiments as well as data on the general performance of the instrument during the USML-2 flight.

  14. Multiparameter Lab-on-a-Chip flow cytometry of the cell cycle.

    PubMed

    Skommer, Joanna; Akagi, Jin; Takeda, Kazuo; Fujimura, Yuu; Khoshmanesh, Khashayar; Wlodkowic, Donald

    2013-04-15

    Multiparameter analysis of apoptosis in relation to cell cycle position is helpful in exploring mechanism of action of anticancer drugs that target specific molecular cogs of the cell cycle. This work demonstrates a new rationale for using microfluidic Lab-on-a-Chip flow cytometry (μFCM) with a simple 2D hydrodynamic focusing for the multiparameter analysis of apoptosis and DNA ploidy analysis in human hematopoietic cancer cells. The microfluidic system employs disposable microfluidic cartridges fabricated using injection moulding in optically transparent poly(methylmethacrylate). The dedicated and miniaturized electronic hardware interface enables up to six parameter detections using a combination of spatially separated solid-state 473 nm (10 mW) and 640 nm (20 mW) lasers and x-y stage for rapid laser alignment adjustment. We provide evidence that the simple 2D flow focusing on a chip-based device is sufficient to measure cellular DNA content in both fixed and living tumor cells. The feasibility of using the μFCM system for multiparameter analysis of caspase activation and dissipation of mitochondrial inner membrane potential (ΔΨ(m) loss) in relation to DNA content is also demonstrated. The data shows that straightforward microfluidic chip designs are sufficient to acquire high quality biological data when combined with sophisticated electronic interfaces. They can be a viable alternative to conventional FCM for multiparameter detection of programmed cell death.

  15. Ultrafast 2D IR microscopy

    PubMed Central

    Baiz, Carlos R.; Schach, Denise; Tokmakoff, Andrei

    2014-01-01

    We describe a microscope for measuring two-dimensional infrared (2D IR) spectra of heterogeneous samples with μm-scale spatial resolution, sub-picosecond time resolution, and the molecular structure information of 2D IR, enabling the measurement of vibrational dynamics through correlations in frequency, time, and space. The setup is based on a fully collinear “one beam” geometry in which all pulses propagate along the same optics. Polarization, chopping, and phase cycling are used to isolate the 2D IR signals of interest. In addition, we demonstrate the use of vibrational lifetime as a contrast agent for imaging microscopic variations in molecular environments. PMID:25089490

  16. Spatial response variations within biosensor flow cells

    NASA Astrophysics Data System (ADS)

    Cant, Nicola; Harrison, Sarah

    2012-02-01

    Biosensors are currently being developed for the detection of a wide range of analytes in a variety of scenarios. One such area is that of environmental monitoring for the presence of biological threats, from toxins through to viruses and bacteria. The varying nature, and in particular disparate size, of such a variety of analytes poses a significant challenge in the development of effective high confidence instruments. Many existing biosensors employ functionalised flow cells in which spatially defined arrays of surface immobilised recognition elements are present to specifically capture their analyte of interest. Experimental data obtained using a grating coupled SPR instrument, the BIAcore Flexchip, has revealed spatial dependency differences in response behaviours between proteinaceous and particulate analytes. In particular, the magnitude of responses seen with Bacillus anthracis spores across the instruments flow cell appear to be influenced by shear and gravitational effects whilst those from soluble proteins are more uniform. We have explored this dependence to understand its fundamental impact on the successful implementation of multi-analyte environmental biological detection systems.

  17. Webcam-based flow cytometer using wide-field imaging for low cell number detection at high throughput.

    PubMed

    Balsam, Joshua; Bruck, Hugh Alan; Rasooly, Avraham

    2014-09-01

    Here we describe a novel low-cost flow cytometer based on a webcam capable of low cell number detection in a large volume which may overcome the limitations of current flow cytometry. Several key elements have been combined to yield both high throughput and high sensitivity. The first element is a commercially available webcam capable of 187 frames per second video capture at a resolution of 320 × 240 pixels. The second element in this design is a 1 W 450 nm laser module for area-excitation, which combined with the webcam allows for rapid interrogation of a flow field. The final element is a 2D flow-cell which overcomes the flow limitation of hydrodynamic focusing and allows for higher sample throughput in a wider flow field. This cell allows for the linear velocity of target cells to be lower than in a conventional "1D" hydrodynamic focusing flow-cells typically used in cytometry at similar volumetric flow rates. It also allows cells to be imaged at the full frame rate of the webcam. Using this webcam-based flow cytometer with wide-field imaging, it was confirmed that the detection of fluorescently tagged 5 μm polystyrene beads in "1D" hydrodynamic focusing flow-cells was not practical for low cell number detection due to streaking from the motion of the beads, which did not occur with the 2D flow-cell design. The sensitivity and throughput of this webcam-based flow cytometer was then investigated using THP-1 human monocytes stained with SYTO-9 florescent dye in the 2D flow-cell. The flow cytometer was found to be capable of detecting fluorescently tagged cells at concentrations as low as 1 cell per mL at flow rates of 500 μL min(-1) in buffer and in blood. The effectiveness of detection was concentration dependent: at 100 cells per mL 84% of the cells were detected compared to microscopy, 10 cells per mL 79% detected and 1 cell per mL 59% of the cells were detected. With the blood samples spiked to 100 cells per mL, the average concentration for all samples

  18. AnisWave 2D

    2004-08-01

    AnisWave2D is a 2D finite-difference code for a simulating seismic wave propagation in fully anisotropic materials. The code is implemented to run in parallel over multiple processors and is fully portable. A mesh refinement algorithm has been utilized to allow the grid-spacing to be tailored to the velocity model, avoiding the over-sampling of high-velocity materials that usually occurs in fixed-grid schemes.

  19. Co-flow planar SOFC fuel cell stack

    DOEpatents

    Chung, Brandon W.; Pham, Ai Quoc; Glass, Robert S.

    2004-11-30

    A co-flow planar solid oxide fuel cell stack with an integral, internal manifold and a casing/holder to separately seal the cell. This construction improves sealing and gas flow, and provides for easy manifolding of cell stacks. In addition, the stack construction has the potential for an improved durability and operation with an additional increase in cell efficiency. The co-flow arrangement can be effectively utilized in other electrochemical systems requiring gas-proof separation of gases.

  20. Phenotyping breast cancer cell lines EM-G3, HCC1937, MCF7 and MDA-MB-231 using 2-D electrophoresis and affinity chromatography for glutathione-binding proteins

    PubMed Central

    2010-01-01

    Background Transformed phenotypes are common to cell lines derived from various cancers. Proteome profiling is a valuable tool that may reveal uncharacteristic cell phenotypes in transformed cells. Changes in expression of glutathione S-transferases (GSTs) and other proteins interacting with glutathione (GSH) in model cell lines could be of particular interest. Methods We compared the phenotypes of breast cell lines EM-G3, HCC1937, MCF7 and MDA-MB-231 using 2-D electrophoresis (2-DE). We further separated GSH-binding proteins from the cell lines using affinity chromatography with GSH-Sepharose 4B, performed 2-DE analysis and identified the main protein spots. Results Correlation coefficients among 2-DE gels from the cell lines were lower than 0.65, pointing to dissimilarity among the cell lines. Differences in primary constituents of the cytoskeleton were shown by the 2-D protein maps and western blots. The spot patterns in gels of GSH-binding fractions from primary carcinoma-derived cell lines HCC1937 and EM-G3 were similar to each other, and they differed from the spot patterns of cell lines MCF7 and MDA-MB-231 that were derived from pleural effusions of metastatic mammary carcinoma patients. Major differences in the expression of GST P1-1 and carbonyl reductase [NADPH] 1 were observed among the cell lines, indicating differential abilities of the cell lines to metabolize xenobiotics. Conclusions Our results confirmed the applicability of targeted affinity chromatography to proteome profiling and allowed us to characterize the phenotypes of four breast cancer cell lines. PMID:20731849

  1. Cancer cell glycocalyx mediates mechanotransduction and flow-regulated invasion.

    PubMed

    Qazi, Henry; Palomino, Rocio; Shi, Zhong-Dong; Munn, Lance L; Tarbell, John M

    2013-11-01

    Mammalian cells are covered by a surface proteoglycan (glycocalyx) layer, and it is known that blood vessel-lining endothelial cells use the glycocalyx to sense and transduce the shearing forces of blood flow into intracellular signals. Tumor cells in vivo are exposed to forces from interstitial fluid flow that may affect metastatic potential but are not reproduced by most in vitro cell motility assays. We hypothesized that glycocalyx-mediated mechanotransduction of interstitial flow shear stress is an un-recognized factor that can significantly enhance metastatic cell motility and play a role in augmentation of invasion. Involvement of MMP levels, cell adhesion molecules (CD44, α3 integrin), and glycocalyx components (heparan sulfate and hyaluronan) was investigated in a cell/collagen gel suspension model designed to mimic the interstitial flow microenvironment. Physiological levels of flow upregulated MMP levels and enhanced the motility of metastatic cells. Blocking the flow-enhanced expression of MMP activity or adhesion molecules (CD44 and integrins) resulted in blocking the flow-enhanced migratory activity. The presence of a glycocalyx-like layer was verified around tumor cells, and the degradation of this layer by hyaluronidase and heparinase blocked the flow-regulated invasion. This study shows for the first time that interstitial flow enhancement of metastatic cell motility can be mediated by the cell surface glycocalyx - a potential target for therapeutics.

  2. MAGNUM-2D computer code: user's guide

    SciTech Connect

    England, R.L.; Kline, N.W.; Ekblad, K.J.; Baca, R.G.

    1985-01-01

    Information relevant to the general use of the MAGNUM-2D computer code is presented. This computer code was developed for the purpose of modeling (i.e., simulating) the thermal and hydraulic conditions in the vicinity of a waste package emplaced in a deep geologic repository. The MAGNUM-2D computer computes (1) the temperature field surrounding the waste package as a function of the heat generation rate of the nuclear waste and thermal properties of the basalt and (2) the hydraulic head distribution and associated groundwater flow fields as a function of the temperature gradients and hydraulic properties of the basalt. MAGNUM-2D is a two-dimensional numerical model for transient or steady-state analysis of coupled heat transfer and groundwater flow in a fractured porous medium. The governing equations consist of a set of coupled, quasi-linear partial differential equations that are solved using a Galerkin finite-element technique. A Newton-Raphson algorithm is embedded in the Galerkin functional to formulate the problem in terms of the incremental changes in the dependent variables. Both triangular and quadrilateral finite elements are used to represent the continuum portions of the spatial domain. Line elements may be used to represent discrete conduits. 18 refs., 4 figs., 1 tab.

  3. Matrine increases NKG2D ligand ULBP2 in K562 cells via inhibiting JAK/STAT3 pathway: a potential mechanism underlying the immunotherapy of matrine in leukemia

    PubMed Central

    Lu, Xuzhang; Zhu, Zhichao; Jiang, Lijia; Sun, Xiao; Jia, Zhuxia; Qian, Sixuan; Li, Jianyong; Ma, Lingdi

    2015-01-01

    Purpose: The study aimed to investigate the role of the JAK/STAT3 pathway in the matrine induced ULBP2 expression on the human chronic myelogenous leukemia K562 cells. Methods: K562 cells were cultured, and the relevant mRNA expressions were detected. Results: Matrine induced the expression of four NKG2D ligands on K562 cells, of which ULBP2 had the highest increase. After treatment with 0.8 mg/mL matrine for 24 h, the mean fluorescence intensity (MFI) of ULBP2 increased. After matrine treatment, the sensitivity of K562 cells to NK cell-mediated killing increased significantly. After treatment with 0.2, 0.5 and 0.8 mg/ mL matrine, the percentage of K562 cells killed by NK cells was significantly higher than that of untreated cells (29.2%) (P<0.05). Matrine significantly inhibit the protein expression of phosphorylated STAT 3 and JAK2. Matrine markedly inhibited the IL-6 expression of K562 cells, and antagonized the IL-6 mediated STAT3 and JAK2 phosphorylation. In addition, matrine enhanced the inhibitory effect of STAT 3 inhibitor on STAT 3 activity. The silencing of STAT expression and inhibition of STAT3 activity significantly up-regulated the ULPB2 expression. Matrine had no effect on the expression of IL-6R and gp130 on K562 cells, the mRNA expression of IL-6R and gp130 increased slightly and the sgp 130 in cell supernatant significantly increased. Conclusions: Our findings reveal IL-6 and IL-6 receptor-mediated JAK/STAT3 pathway is involved in the matrine induced up-regulation of NKG2D ligands ULBP2 on K562 cells. Matrine might inhibit IL-6 expression and then suppress the activation of IL-6 receptor-mediated JAK/STAT3 pathway. PMID:26692928

  4. Investigations on proton exchange membrane fuel cells with different configurations and flow fields

    NASA Astrophysics Data System (ADS)

    Kazim, Ayoub Mohamed

    In this study, two mathematical models are developed. The first one is a simple mathematical approach that computes all transport and electrochemical parameters inside the different layers of a fuel cell regardless of its configuration. Through heat and mass transfer analogy, convective mass transfer coefficients at different Reynolds number are determined for both concentric cylindrical and conventional proton exchange membrane (PEM) fuel cells. Concentrations of oxygen and hydrogen are then determined at each layer of the fuel cell using steady-state diffusion analysis. The concentration equations are solved together with the electrochemical equations inside the fuel cell, to obtain the fuel cell voltage and power density. The results from this simple approach compared well with the existing numerical and experimental results. The second mathematical model is to study PEM fuel cell with conventional and non-conventional namely interdigitated flow fields. Through proper handling of the boundary conditions at the gas diffusion/catalyst layer interface, the numerical solution of the model resulted in the profiles of transport and electrochemical parameters in the cathode. Parameters such as pressure distribution, velocity profile, oxygen concentration, molar flux, current density, polarization and overall power density at different cell over-potentials in both flow fields were determined. The results demonstrates the superiority of interdigitated flow field over the conventional type in terms of overall performance and illustrated the importance of the convective term of the species equation in enhancing the reaction rates, leading to a significant improvement in the fuel cell performance. The effects of different parameters, such as cathode porosity, inlet oxygen mole fraction, and operating pressure on fuel cell performance have been studied using this 2-D mathematical model. Finally, a simple efficiency and economical analysis was formulated and implemented on

  5. A modular segmented-flow platform for 3D cell cultivation.

    PubMed

    Lemke, Karen; Förster, Tobias; Römer, Robert; Quade, Mandy; Wiedemeier, Stefan; Grodrian, Andreas; Gastrock, Gunter

    2015-07-10

    In vitro 3D cell cultivation is promised to equate tissue in vivo more realistically than 2D cell cultivation corresponding to cell-cell and cell-matrix interactions. Therefore, a scalable 3D cultivation platform was developed. This platform, called pipe-based bioreactors (pbb), is based on the segmented-flow technology: aqueous droplets are embedded in a water-immiscible carrier fluid. The droplet volumes range from 60 nL to 20 μL and are used as bioreactors lined up in a tubing like pearls on a string. The modular automated platform basically consists of several modules like a fluid management for a high throughput droplet generation for self-assembly or scaffold-based 3D cell cultivation, a storage module for incubation and storage, and an analysis module for monitoring cell aggregation and proliferation basing on microscopy or photometry. In this report, the self-assembly of murine embryonic stem cells (mESCs) to uniformly sized embryoid bodies (EBs), the cell proliferation, the cell viability as well as the influence on the cell differentiation to cardiomyocytes are described. The integration of a dosage module for medium exchange or agent addition will enable pbb as long-term 3D cell cultivation system for studying stem cell differentiation, e.g. cardiac myogenesis or for diagnostic and therapeutic testing in personalized medicine.

  6. Breakthrough Flow Battery Cell Stack: Transformative Electrochemical Flow Storage System (TEFSS)

    SciTech Connect

    2010-09-09

    GRIDS Project: UTRC is developing a flow battery with a unique design that provides significantly more power than today's flow battery systems. A flow battery is a cross between a traditional battery and a fuel cell. Flow batteries store their energy in external tanks instead of inside the cell itself. Flow batteries have traditionally been expensive because the battery cell stack, where the chemical reaction takes place, is costly. In this project, UTRC is developing a new stack design that achieves 10 times higher power than today’s flow batteries. This high power output means the size of the cell stack can be smaller, reducing the amount of expensive materials that are needed. UTRC’s flow battery will reduce the cost of storing electricity for the electric grid, making widespread use feasible.

  7. Counter-Flow Cooling Tower Test Cell

    NASA Astrophysics Data System (ADS)

    Dvořák, Lukáš; Nožička, Jiří

    2014-03-01

    The article contains a design of a functional experimental model of a cross-flow mechanical draft cooling tower and the results and outcomes of measurements. This device is primarily used for measuring performance characteristics of cooling fills, but with a simple rebuild, it can be used for measuring other thermodynamic processes that take part in so-called wet cooling. The main advantages of the particular test cell lie in the accuracy, size, and the possibility of changing the water distribution level. This feature is very useful for measurements of fills of different heights without the influence of the spray and rain zone. The functionality of this test cell has been verified experimentally during assembly, and data from the measurement of common film cooling fills have been compared against the results taken from another experimental line. For the purpose of evaluating the data gathered, computational scripts were created in the MATLAB numerical computing environment. The first script is for exact calculation of the thermal balance of the model, and the second is for determining Merkel's number via Chebyshev's method.

  8. Aqueous semi-solid flow cell: demonstration and analysis.

    PubMed

    Li, Zheng; Smith, Kyle C; Dong, Yajie; Baram, Nir; Fan, Frank Y; Xie, Jing; Limthongkul, Pimpa; Carter, W Craig; Chiang, Yet-Ming

    2013-10-14

    An aqueous Li-ion flow cell using suspension-based flow electrodes based on the LiTi2(PO4)3-LiFePO4 couple is demonstrated. Unlike conventional flow batteries, the semi-solid approach utilizes fluid electrodes that are electronically conductive. A model of simultaneous advection and electrochemical transport is developed and used to separate flow-induced losses from those due to underlying side reactions. The importance of plug flow to achieving high energy efficiency in flow batteries utilizing highly non-Newtonian flow electrodes is emphasized.

  9. Aqueous semi-solid flow cell: demonstration and analysis

    SciTech Connect

    Li, Z; Smith, KC; Dong, YJ; Baram, N; Fan, FY; Xie, J; Limthongkul, P; Carter, WC; Chiang, YM

    2013-01-01

    An aqueous Li-ion flow cell using suspension-based flow electrodes based on the LiTi2(PO4)(3)-LiFePO4 couple is demonstrated. Unlike conventional flow batteries, the semi-solid approach utilizes fluid electrodes that are electronically conductive. A model of simultaneous advection and electrochemical transport is developed and used to separate flow-induced losses from those due to underlying side reactions. The importance of plug flow to achieving high energy efficiency in flow batteries utilizing highly non-Newtonian flow electrodes is emphasized.

  10. 1,25(OH)2D3 and VDR Signaling Pathways Regulate the Inhibition of Dectin-1 Caused by Cyclosporine A in Response to Aspergillus Fumigatus in Human Corneal Epithelial Cells

    PubMed Central

    Xia, Yiping; Zhao, Guiqiu; Lin, Jing; Li, Cui; Cong, Lin; Jiang, Nan; Xu, Qiang; Wang, Qian

    2016-01-01

    Background The objective of this study is to observe whether cyclosporine A (CsA) inhibits the expression of dectin-1 in human corneal epithelial cells infected with Aspergillus fumigatus (A. fumigatus) and to investigate the molecular mechanisms of the inhibition. Methods Immortalized human corneal epithelial cells (HCECs) were pretreated with 1,25(OH)2D3 and VDR inhibitor for 1 h, and then they were pretreated with CsA for 12h. After these pretreatments, the HCECs were stimulated with A. fumigatus and curdlan respectively, and the expression of dectin-1 and proinflammatory cytokines (IL-1β and TNF-α) were detected by RT-PCR, western blot and ELISA. Results Dectin-1 mRNA and dectin-1 protein expression increased when HCECs were stimulated with A. fumigatus or curdlan, and CsA inhibited the dectin-1 expression both in mRNA and protein levels specifically. Dectin-1 and proinflammatory cytokine expression levels were higher when HCECs were pretreated with VDR inhibitor and CsA compared to pretreatment with CsA alone, while dectin-1 and proinflammatory cytokine levels were lower when HCECs were pretreated with 1,25(OH)2D3 and CsA compared to pretreatment with CsA alone. Conclusions These data provide evidence that CsA can inhibit the expression of dectin-1 and proinflammatory cytokines through dectin-1 when HCECs are stimulated by A. fumigatus or curdlan. The active form of vitamin D, 1,25(OH)2D3, and VDR signaling pathway regulate the inhibition of CsA. The inhibition is enhanced by 1,25(OH)2D3, and the VDR inhibitor suppresses the inhibition. PMID:27755569

  11. Traditional Chinese medicine herbal mixture LQ arrests FUCCI-expressing HeLa cells in G₀/G₁ phase in 2D plastic, 2.5D Matrigel, and 3D Gelfoam culture visualized with FUCCI imaging.

    PubMed

    Zhang, Lei; Wu, Chengyu; Bouvet, Michael; Yano, Shuya; Hoffman, Robert M

    2015-03-10

    We used the fluorescence ubiquitination-based cell cycle indicator (FUCCI) to monitor cell cycle arrest after treatment of FUCCI-expressing HeLa cells (FUCCI-HeLa) with a traditional Chinese medicine (TCM) herbal mixture LQ, previously shown to have anti-tumor and anti-metastatic activity in mouse models. Paclitaxel was used as the positive control. In 2D monolayer culture, the untreated control had approximately 45% of the cells in S/G₂/M phase. In contrast, the LQ-treated cells (9 mg/ml) were mostly in the G₀/G₁ (>90%) after 72 hours. After treatment with paclitaxel (0.01 μm), for 72 hours, 95% of the cells were in S/G₂/M. In 2.5D Matrigel culture, the colonies in the untreated control group had 40% of the cells in S/G₂/M. LQ arrested the cells in G₀/G₁ after 72 hours. Paclitaxel arrested almost all the cells in S/G₂/M after 72 hours. In 3D Gelfoam culture, the untreated control culture had approximately 45% of cells in G₂/M. In contrast, the LQ-treated cells were mostly in G₀/G₁ phase (>80%) after 72 hours treatment. Paclitaxel resulted in 90% of the cells arrested in S/G₂/M after 72 hours. The present report suggests the non-toxic LQ has potential to maintain cancers in a quiescent state for long periods of time.

  12. Label free cell-tracking and division detection based on 2D time-lapse images for lineage analysis of early embryo development.

    PubMed

    Cicconet, Marcelo; Gutwein, Michelle; Gunsalus, Kristin C; Geiger, Davi

    2014-08-01

    In this paper we report a database and a series of techniques related to the problem of tracking cells, and detecting their divisions, in time-lapse movies of mammalian embryos. Our contributions are (1) a method for counting embryos in a well, and cropping each individual embryo across frames, to create individual movies for cell tracking; (2) a semi-automated method for cell tracking that works up to the 8-cell stage, along with a software implementation available to the public (this software was used to build the reported database); (3) an algorithm for automatic tracking up to the 4-cell stage, based on histograms of mirror symmetry coefficients captured using wavelets; (4) a cell-tracking database containing 100 annotated examples of mammalian embryos up to the 8-cell stage; and (5) statistical analysis of various timing distributions obtained from those examples. PMID:24873887

  13. Label Free Cell-Tracking and Division Detection Based on 2D Time-Lapse Images For Lineage Analysis of Early Embryo Development

    PubMed Central

    Cicconet, Marcelo; Gutwein, Michelle; Gunsalus, Kristin C; Geiger, Davi

    2014-01-01

    In this paper we report a database and a series of techniques related to the problem of tracking cells, and detecting their divisions, in time-lapse movies of mammalian embryos. Our contributions are: (1) a method for counting embryos in a well, and cropping each individual embryo across frames, to create individual movies for cell tracking; (2) a semi-automated method for cell tracking that works up to the 8-cell stage, along with a software implementation available to the public (this software was used to build the reported database); (3) an algorithm for automatic tracking up to the 4-cell stage, based on histograms of mirror symmetry coefficients captured using wavelets; (4) a cell-tracking database containing 100 annotated examples of mammalian embryos up to the 8-cell stage; (5) statistical analysis of various timing distributions obtained from those examples. PMID:24873887

  14. DYNA2D96. Explicit 2-D Hydrodynamic FEM Program

    SciTech Connect

    Whirley, R.G.

    1992-04-01

    DYNA2D is a vectorized, explicit, two-dimensional, axisymmetric and plane strain finite element program for analyzing the large deformation dynamic and hydrodynamic response of inelastic solids. DYNA2D contains 13 material models and 9 equations of state (EOS) to cover a wide range of material behavior. The material models implemented in all machine versions are: elastic, orthotropic elastic, kinematic/isotropic elastic plasticity, thermoelastoplastic, soil and crushable foam, linear viscoelastic, rubber, high explosive burn, isotropic elastic-plastic, temperature-dependent elastic-plastic. The isotropic and temperature-dependent elastic-plastic models determine only the deviatoric stresses. Pressure is determined by one of 9 equations of state including linear polynomial, JWL high explosive, Sack Tuesday high explosive, Gruneisen, ratio of polynomials, linear polynomial with energy deposition, ignition and growth of reaction in HE, tabulated compaction, and tabulated.

  15. ( sup 3 H)(D-PEN sup 2 , D-PEN sup 5 ) enkephalin binding to delta opioid receptors on intact neuroblastoma-glioma (NG 108-15) hybrid cells

    SciTech Connect

    Knapp, R.J.; Yamamura, H.I. )

    1990-01-01

    ({sup 3}H)(D-Pen{sup 2}, D-Pen{sup 5})enkephalin binding to intact NG 108-15 cells has been measured under physiological conditions of temperature and medium. The dissociation constant, receptor density, and Hill slope values measured under these conditions are consistent with values obtained by others using membranes prepared from these cells. Kinetic analysis of the radioligand binding to these cells show biphasic association and monophasic dissociation processes suggesting the presence of different receptor affinity states for the agonist. The data show that the binding affinity of ({sup 3}H)(D-Pen{sup 2}, D-Pen{sup 5})enkephalin under physiological conditions is not substantially different to that measured in 50 mM Tris buffer using cell membrane fractions. Unlike DPDPE, the {mu} opioid agonists morphine, normorphine, PL-17, and DAMGO, have much lower affinity for the {delta} receptor measured under these conditions than is observed by studies using 50 mM Tris buffer. The results described here suggest that this assay may serve as a useful model of {delta} opioid receptor binding in vivo.

  16. Ultrasonic Enrichment of Flowing Blood Cells in Capillars: Influence of the Flow Rate

    NASA Astrophysics Data System (ADS)

    Carreras, Pilar; Gonzalez, Itziar; Ahumada, Oscar

    Red blood cells subjected to standing waves collect at the pressure nodes during their flow motion. Blood is a non-newtonian fluid whose density and other properties are defined by its flow velocity. Their drift motion is governed by the radiation force together with hydrodynamic conditions. This work presents a study of the blood cell enrichment performed in a rectangular capillar at f=1 MHz as a function of their flow motion. The cells collect along the central axis of the capillary in very few seconds, with a clearance in other lateral areas. Optimal flow rates below 100uL/min were found in the experiments.

  17. Parallel-plate flow chamber and continuous flow circuit to evaluate endothelial progenitor cells under laminar flow shear stress.

    PubMed

    Lane, Whitney O; Jantzen, Alexandra E; Carlon, Tim A; Jamiolkowski, Ryan M; Grenet, Justin E; Ley, Melissa M; Haseltine, Justin M; Galinat, Lauren J; Lin, Fu-Hsiung; Allen, Jason D; Truskey, George A; Achneck, Hardean E

    2012-01-01

    The overall goal of this method is to describe a technique to subject adherent cells to laminar flow conditions and evaluate their response to well quantifiable fluid shear stresses. Our flow chamber design and flow circuit (Fig. 1) contains a transparent viewing region that enables testing of cell adhesion and imaging of cell morphology immediately before flow (Fig. 11A, B), at various time points during flow (Fig. 11C), and after flow (Fig. 11D). These experiments are illustrated with human umbilical cord blood-derived endothelial progenitor cells (EPCs) and porcine EPCs. This method is also applicable to other adherent cell types, e.g. smooth muscle cells (SMCs) or fibroblasts. The chamber and all parts of the circuit are easily sterilized with steam autoclaving. In contrast to other chambers, e.g. microfluidic chambers, large numbers of cells (> 1 million depending on cell size) can be recovered after the flow experiment under sterile conditions for cell culture or other experiments, e.g. DNA or RNA extraction, or immunohistochemistry (Fig. 11E), or scanning electron microscopy. The shear stress can be adjusted by varying the flow rate of the perfusate, the fluid viscosity, or the channel height and width. The latter can reduce fluid volume or cell needs while ensuring that one-dimensional flow is maintained. It is not necessary to measure chamber height between experiments, since the chamber height does not depend on the use of gaskets, which greatly increases the ease of multiple experiments. Furthermore, the circuit design easily enables the collection of perfusate samples for analysis and/or quantification of metabolites secreted by cells under fluid shear stress exposure, e.g. nitric oxide (Fig. 12).

  18. Soluble MICA-NKG2D interaction upregulates IFN-γ production by activated CD3-CD56+ NK cells: potential impact on chronic graft versus host disease.

    PubMed

    Boukouaci, Wahid; Al-Daccak, Reem; Dulphy, Nicolas; Lauden, Laura; Amokrane, Kahina; Fortier, Catherine; Marzais, François; Bennabi, Meriem; Peffault de Latour, Regis; Socie, Gerard; Toubert, Antoine; Charron, Dominique; Krishnamoorthy, Rajagopal; Tamouza, Ryad

    2013-12-01

    A soluble isoform of MHC class I chain-related molecule A (soluble MICA), generated by proteolytic shedding from the membrane-bound MICA of various tumor cells, has been shown to downregulate both the expression of natural killer group 2-member D receptor and the cytotoxic function of effectors cells and was postulated as a mechanism for tumor immune evasion. Its effect on the expression of cytokines by the effector cells remained unexplored. Here we demonstrate that the sMICA molecules upregulate interferon gamma expression by interleukin-12/interleukin-18-activated CD3(-)CD56(+) natural killer cells, witnessing the pro-inflammatory effect of soluble MICA. Overall, these data are in line with our previous observations that the raised serum levels of soluble MICA, following allogeneic hematopoietic stem cell transplantation, confer susceptibility to and the presence of pre-transplantation anti-MICA antibodies in the patient's serum confer protection against chronic graft versus host disease.

  19. Mechanical loading and the synthesis of 1,25(OH)2D in primary human osteoblasts.

    PubMed

    van der Meijden, K; Bakker, A D; van Essen, H W; Heijboer, A C; Schulten, E A J M; Lips, P; Bravenboer, N

    2016-02-01

    The metabolite 1,25-dihydroxyvitamin D (1,25(OH)2D) is synthesized from its precursor 25-hydroxyvitamin D (25(OH)D) by human osteoblasts leading to stimulation of osteoblast differentiation in an autocrine or paracrine way. Osteoblast differentiation is also stimulated by mechanical loading through activation of various responses in bone cells such as nitric oxide signaling. Whether mechanical loading affects osteoblast differentiation through an enhanced synthesis of 1,25(OH)2D by human osteoblasts is still unknown. We hypothesized that mechanical loading stimulates the synthesis of 1,25(OH)2D from 25(OH)D in primary human osteoblasts. Since the responsiveness of bone to mechanical stimuli can be altered by various endocrine factors, we also investigated whether 1,25(OH)2D or 25(OH)D affect the response of primary human osteoblasts to mechanical loading. Primary human osteoblasts were pre-incubated in medium with/without 25(OH)D3 (400 nM) or 1,25(OH)2D3 (100 nM) for 24h and subjected to mechanical loading by pulsatile fluid flow (PFF). The response of osteoblasts to PFF was quantified by measuring nitric oxide, and by PCR analysis. The effect of PFF on the synthesis of 1,25(OH)2D3 was determined by subjecting osteoblasts to PFF followed by 24h post-incubation in medium with/without 25(OH)D3 (400 nM). We showed that 1,25(OH)2D3 reduced the PFF-induced NO response in primary human osteoblasts. 25(OH)D3 did not significantly alter the NO response of primary human osteoblasts to PFF, but 25(OH)D3 increased osteocalcin and RANKL mRNA levels, similar to 1,25(OH)2D3. PFF did not increase 1,25(OH)2D3 amounts in our model, even though PFF did increase CYP27B1 mRNA levels and reduced VDR mRNA levels. CYP24 mRNA levels were not affected by PFF, but were strongly increased by both 25(OH)D3 and 1,25(OH)2D3. In conclusion, 1,25(OH)2D3 may affect the response of primary human osteoblasts to mechanical stimuli, at least with respect to NO production. Mechanical stimuli may affect

  20. Traction Forces of Endothelial Cells under Slow Shear Flow

    PubMed Central

    Perrault, Cecile M.; Brugues, Agusti; Bazellieres, Elsa; Ricco, Pierre; Lacroix, Damien; Trepat, Xavier

    2015-01-01

    Endothelial cells are constantly exposed to fluid shear stresses that regulate vascular morphogenesis, homeostasis, and disease. The mechanical responses of endothelial cells to relatively high shear flow such as that characteristic of arterial circulation has been extensively studied. Much less is known about the responses of endothelial cells to slow shear flow such as that characteristic of venous circulation, early angiogenesis, atherosclerosis, intracranial aneurysm, or interstitial flow. Here we used a novel, to our knowledge, microfluidic technique to measure traction forces exerted by confluent vascular endothelial cell monolayers under slow shear flow. We found that cells respond to flow with rapid and pronounced increases in traction forces and cell-cell stresses. These responses are reversible in time and do not involve reorientation of the cell body. Traction maps reveal that local cell responses to slow shear flow are highly heterogeneous in magnitude and sign. Our findings unveil a low-flow regime in which endothelial cell mechanics is acutely responsive to shear stress. PMID:26488643

  1. Numerical simulations of cell interactions under shear flows in complex geometries

    NASA Astrophysics Data System (ADS)

    Peng, Gaozhu

    2005-11-01

    The receptor-mediated leukocyte adhesion and rolling on endothelium under shear flows are of crucial importance in governing a range of cell functions: inflammatory response, lymphocyte homing, and sickle cell vascular occlusion. In vivo, an endothelium-lined blood vessel lumen has a non-flat irregular complex geometry presented to blood flows, and adherent leukocytes can lead to further geometry complexity. This geometry factor can have a prominent impact on the mechanics and hemodynamics of cell interactions and adhesions in high endothelial venules, non-uniform capillaries and post-capillary expansions to name a few. In this work, a ghost-cell immerse boundary/front tracking method is presented to examine the physiological role of the blood vessel geometry in microcirculation. Motions of deformable blood cells are computed via a multiphase front tracking method. Boundary conditions for arbitrary geometries are enforced through a high-order ghost cell immersed boundary method. The current method is validated and used to explore the potential roles of vessel geometry in modulating hemodynamics and kinetics of 2d/3d cell interactions, in particular leukocyte adhesion and accumulation.

  2. MOSS2D V1

    2001-01-31

    This software reduces the data from two-dimensional kSA MOS program, k-Space Associates, Ann Arbor, MI. Initial MOS data is recorded without headers in 38 columns, with one row of data per acquisition per lase beam tracked. The final MOSS 2d data file is reduced, graphed, and saved in a tab-delimited column format with headers that can be plotted in any graphing software.

  3. Flow and Diffusion in Channel-Guided Cell Migration

    PubMed Central

    Marel, Anna-Kristina; Zorn, Matthias; Klingner, Christoph; Wedlich-Söldner, Roland; Frey, Erwin; Rädler, Joachim O.

    2014-01-01

    Collective migration of mechanically coupled cell layers is a notable feature of wound healing, embryonic development, and cancer progression. In confluent epithelial sheets, the dynamics have been found to be highly heterogeneous, exhibiting spontaneous formation of swirls, long-range correlations, and glass-like dynamic arrest as a function of cell density. In contrast, the flow-like properties of one-sided cell-sheet expansion in confining geometries are not well understood. Here, we studied the short- and long-term flow of Madin-Darby canine kidney (MDCK) cells as they moved through microchannels. Using single-cell tracking and particle image velocimetry (PIV), we found that a defined averaged stationary cell current emerged that exhibited a velocity gradient in the direction of migration and a plug-flow-like profile across the advancing sheet. The observed flow velocity can be decomposed into a constant term of directed cell migration and a diffusion-like contribution that increases with density gradient. The diffusive component is consistent with the cell-density profile and front propagation speed predicted by the Fisher-Kolmogorov equation. To connect diffusion-mediated transport to underlying cellular motility, we studied single-cell trajectories and occurrence of vorticity. We discovered that the directed large-scale cell flow altered fluctuations in cellular motion at short length scales: vorticity maps showed a reduced frequency of swirl formation in channel flow compared with resting sheets of equal cell density. Furthermore, under flow, single-cell trajectories showed persistent long-range, random-walk behavior superimposed on drift, whereas cells in resting tissue did not show significant displacements with respect to neighboring cells. Our work thus suggests that active cell migration manifests itself in an underlying, spatially uniform drift as well as in randomized bursts of short-range correlated motion that lead to a diffusion-mediated transport

  4. Targeted fluorescence imaging enhanced by 2D materials: a comparison between 2D MoS2 and graphene oxide.

    PubMed

    Xie, Donghao; Ji, Ding-Kun; Zhang, Yue; Cao, Jun; Zheng, Hu; Liu, Lin; Zang, Yi; Li, Jia; Chen, Guo-Rong; James, Tony D; He, Xiao-Peng

    2016-08-01

    Here we demonstrate that 2D MoS2 can enhance the receptor-targeting and imaging ability of a fluorophore-labelled ligand. The 2D MoS2 has an enhanced working concentration range when compared with graphene oxide, resulting in the improved imaging of both cell and tissue samples.

  5. Can anti-migratory drugs be screened in vitro? A review of 2D and 3D assays for the quantitative analysis of cell migration.

    PubMed

    Decaestecker, Christine; Debeir, Olivier; Van Ham, Philippe; Kiss, Robert

    2007-03-01

    The aim of the present review is to detail and analyze the pros and cons of in vitro tests available to quantify the anti-migratory effects of anti-cancer drugs for their eventual use in combating the dispersal of tumor cells, a clinical need which currently remains unsatisfied. We therefore briefly sum up why anti-migratory drugs constitute a promising approach in oncology while at the same time emphasizing that migrating cancer cells are resistant to apoptosis. To analyze the pros and cons of the various in vitro tests under review we also briefly sum up the molecular and cellular stages of cancer cell migration, an approach that enables us to argue both that no single in vitro test is sufficient to characterize the anti-migratory potential of a drug and that standardization is needed for the efficient quantitative analysis of cell locomotion in a 3D environment. Before concluding our review we devote the final two parts (i) to the description of new prototypes which, in the near future, could enter the screening process with a view to identifying novel anti-migratory compounds, and (ii) to the anti-migratory compounds currently developed against cancer, with particular emphasis on how these compounds were selected before entering the clinical trial phase.

  6. Modeling of flow field in polymer electrolyte membrane fuel cell

    NASA Astrophysics Data System (ADS)

    Karvonen, Suvi; Hottinen, Tero; Saarinen, Jaakko; Himanen, Olli

    Isothermal two- and three-dimensional polymer electrolyte membrane (PEM) fuel cell cathode flow field models were implemented to study the behavior of reactant and reaction product gas flow in a parallel channel flow field. The focus was on the flow distribution across the channels and the total pressure drop across the flow field. The effect of the density and viscosity variation in the gas resulting from the composition change due to cell reactions was studied and the models were solved with governing equations based on three different approximations. The focus was on showing how a uniform flow profile can be achieved by improving an existing channel design. The modeling results were verified experimentally. A close to uniform flow distribution was achieved in a parallel channel system.

  7. Research development of designing flow cells for optical absorption detectors.

    PubMed

    Yang, Sandong; Tang, Tao; Li, Tong; Wang, Fengyun; Hao, Qingli

    2016-02-01

    The optical absorption detector is one of the most commonly used detectors for high performance liquid chromatography (HPLC). As a core part of this kind of detector, the designs of flow cells, where light passes through samples for acquiring samples information, will affect the performance of a detector. In order to enhance the signal to noise ratio of detectors and reduce the bands broadening that come from flow cells, it is necessary to design a flow cell with a longer optical path length and a less cell volume while maintaining the luminous flux. However the limitations of the machining capacity make it difficult to increase the optical path length, reduce the cell volume and keep or increase the luminous flux simultaneously. It is a challenge to optimize the designing and machining of flow cells so as to improve the performance of detectors. This review discusses the development of designing flow cells based on the detection principle in some aspects of increasing the optical path length, reducing the cell volume, taking the advantages of total reflection and so on. At the same time, some of the designs are illustrated in detail. These various ideas and structures are significant references for designing flow cells and developing optical absorption detectors. PMID:27382716

  8. New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept

    PubMed Central

    Nava-Castro, Karen; Hernández-Bello, Romel; Muñiz-Hernández, Saé; Escobedo, Galileo; Morales-Montor, Jorge

    2011-01-01

    In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells) is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells. PMID:22187522

  9. Analysis of branched DNA replication and recombination intermediates from prokaryotic cells by two-dimensional (2D) native-native agarose gel electrophoresis.

    PubMed

    Robinson, Nicholas P

    2013-01-01

    Branched DNA molecules are generated by the essential processes of replication and recombination. Owing to their distinctive extended shapes, these intermediates migrate differently from linear double-stranded DNA under certain electrophoretic conditions. However, these branched species exist in the cell at much low abundance than the bulk linear DNA. Consequently, branched molecules cannot be visualized by conventional electrophoresis and ethidium bromide staining. Two-dimensional native-native agarose electrophoresis has therefore been developed as a method to facilitate the separation and visualization of branched replication and recombination intermediates. A wide variety of studies have employed this technique to examine branched molecules in eukaryotic, archaeal, and bacterial cells, providing valuable insights into how DNA is duplicated and repaired in all three domains of life.

  10. Engineering light outcoupling in 2D materials.

    PubMed

    Lien, Der-Hsien; Kang, Jeong Seuk; Amani, Matin; Chen, Kevin; Tosun, Mahmut; Wang, Hsin-Ping; Roy, Tania; Eggleston, Michael S; Wu, Ming C; Dubey, Madan; Lee, Si-Chen; He, Jr-Hau; Javey, Ali

    2015-02-11

    When light is incident on 2D transition metal dichalcogenides (TMDCs), it engages in multiple reflections within underlying substrates, producing interferences that lead to enhancement or attenuation of the incoming and outgoing strength of light. Here, we report a simple method to engineer the light outcoupling in semiconducting TMDCs by modulating their dielectric surroundings. We show that by modulating the thicknesses of underlying substrates and capping layers, the interference caused by substrate can significantly enhance the light absorption and emission of WSe2, resulting in a ∼11 times increase in Raman signal and a ∼30 times increase in the photoluminescence (PL) intensity of WSe2. On the basis of the interference model, we also propose a strategy to control the photonic and optoelectronic properties of thin-layer WSe2. This work demonstrates the utilization of outcoupling engineering in 2D materials and offers a new route toward the realization of novel optoelectronic devices, such as 2D LEDs and solar cells.

  11. Refinement Of Hexahedral Cells In Euler Flow Computations

    NASA Technical Reports Server (NTRS)

    Melton, John E.; Cappuccio, Gelsomina; Thomas, Scott D.

    1996-01-01

    Topologically Independent Grid, Euler Refinement (TIGER) computer program solves Euler equations of three-dimensional, unsteady flow of inviscid, compressible fluid by numerical integration on unstructured hexahedral coordinate grid refined where necessary to resolve shocks and other details. Hexahedral cells subdivided, each into eight smaller cells, as needed to refine computational grid in regions of high flow gradients. Grid Interactive Refinement and Flow-Field Examination (GIRAFFE) computer program written in conjunction with TIGER program to display computed flow-field data and to assist researcher in verifying specified boundary conditions and refining grid.

  12. Single chamber fuel cells: Flow geometry, rate and composition considerations

    SciTech Connect

    Stefan, Ionel C.; Jacobson, Craig P.; Visco, Steven J.; De Jonghe, Lutgard C.

    2003-11-17

    Four different single chamber fuel cell designs were compared using propane-air gas mixtures. Gas flow around the electrodes has a significant influence on the open circuit voltage and the power density of the cell. The strong influence of flow geometry is likely due to its effect on gas composition, particularly on the oxygen chemical potential at the two electrodes as a result of gas mixing. The chamber design which exposes the cathode first to the inlet gas was found to yield the best performance at lower flow rates, while the open tube design with the electrodes equally exposed to the inlet gas worked best at higher flow rates.

  13. Antibodies directed to the gram-negative bacterium Neisseria gonorrhoeae cross-react with the 60 kDa heat shock protein and lead to impaired neurite outgrowth in NTera2/D1 cells.

    PubMed

    Reuss, B; Asif, A R

    2014-09-01

    Children of mothers with prenatal gonococcal infections are of increased risk to develop schizophrenic psychosis in later life. The present study hypothesizes an autoimmune mechanism for this, investigating interactions of a commercial rabbit antiserum directed to Neisseria gonorrhoeae (α-NG) with human NTera2/D1 cells, an established in vitro model for human neuronal differentiation. Immunocytochemistry demonstrated α-NG to label antigens on an intracellular organelle, which by Western blot analysis showed a molecular weight shortly below 72 kDa. An antiserum directed to Neisseria meningitidis (α-NM) reacts with an antigen shortly below 95 kDa, confirming antibody specificity of these interactions. Two-dimensional gel electrophoresis and partial Western transfer, allowed to localize an α-NG reactive protein spot which was identified by LC-Q-TOF MS/MS analysis as mitochondrial heat shock protein Hsp60. This was confirmed by Western blot analysis of α-NG immunoreactivity with a commercial Hsp60 protein sample, with which α-NM failed to interact. Finally, analysis of neurite outgrowth in retinoic acid-stimulated differentiating NTera2-D1 cells, demonstrates that α-NG but not α-NM treatment reduces neurite length. These results demonstrate that α-NG can interact with Hsp60 in vitro, whereas pathogenetic relevance of this interaction for psychotic symptomatology remains to be clarified. PMID:24577885

  14. Numerical Simulation of Sickle Cell Blood Flow in the Microcirculation

    NASA Astrophysics Data System (ADS)

    Berger, Stanley A.; Carlson, Brian E.

    2001-11-01

    A numerical simulation of normal and sickle cell blood flow through the transverse arteriole-capillary microcirculation is carried out to model the dominant mechanisms involved in the onset of vascular stasis in sickle cell disease. The transverse arteriole-capillary network is described by Strahler's network branching method, and the oxygen and blood transport in the capillaries is modeled by a Krogh cylinder analysis utilizing Lighthill's lubrication theory, as developed by Berger and King. Poiseuille's law is used to represent blood flow in the arterioles. Applying this flow and transport model and utilizing volumetric flow continuity at each network bifurcation, a nonlinear system of equations is obtained, which is solved iteratively using a steepest descent algorithm coupled with a Newton solver. Ten different networks are generated and flow results are calculated for normal blood and sickle cell blood without and with precapillary oxygen loss. We find that total volumetric blood flow through the network is greater in the two sickle cell blood simulations than for normal blood owing to the anemia associated with sickle cell disease. The percentage of capillary blockage in the network increases dramatically with decreasing pressure drop across the network in the sickle cell cases while there is no blockage when normal blood flows through simulated networks. It is concluded that, in sickle cell disease, without any vasomotor dilation response to decreasing oxygen concentrations in the blood, capillary blockage will occur in the microvasculature even at average pressure drops across the transverse arteriole-capillary networks.

  15. Laser rastering flow cytometry: fast cell counting and identification

    NASA Astrophysics Data System (ADS)

    Vacca, G.; Junnarkar, M. R.; Goldblatt, N. R.; Yee, M. W.; Van Slyke, B. M.; Briese, T. C.

    2009-02-01

    We describe the concept of laser rastering flow cytometry, where a rapidly scanning laser beam allows counting and classification of cells at much higher rates than currently possible. Modifications to existing flow cytometers to implement the concept include an acousto-optic deflector, fast analog-to-digital conversion, and a two-step digital-signal-processing scheme that handles the high data rates and provides key assay information. Results are shown that prove the concept, demonstrating the ability to resolve closely spaced cells and to measure cells at rates more than an order of magnitude faster than on conventional flow-cytometer-based hematology analyzers.

  16. In situ spazialisation of flow laws in a fractured media with multi-parametric measurements (pressure, deformation, water conductivity) and 2d coupled hydro-mechanical (h.m) modelling.

    NASA Astrophysics Data System (ADS)

    Charmoille, A.; Cappa, F.; Guglielmie, Y.

    2003-04-01

    decreasing. The simultaneous record of deformations in the drain and joint show us that joint deformation is effective after the drain opening. It is necessarily to have a sufficient water pressure in the drain to open the joint, thus the joint deformation is strongly dependent of the water pressure in the drain. When the pressure decreases in the drain, the stress stopped on the joint, and it happens a reduction of the joint aperture which induces a pressure fall less fast than the pressure fall in the drain. In the very low permeable area of the joint, it only happens a lightly decrease of pressure. In this investigation area, the joint opening is less important than in the low permeable area. During the pressure decrease in the drain, the joint closed, and the new affected permeability only permits water to slowly flow out of the joint. Then, we induce the second pressure increase when the pressure decrease in this area doesn’t finish. The reopening of the joint with the arrivals of water induces a new pressure increase which adds to the residual pressure, thus, it happens an accumulation of pressure in the joint. This observation shows us that the flow in the fractured media isn’t governed by the same parameters in a low permeable discontinuity or in a very permeable discontinuity. The conductivity measurements in the joint and in drain clearly highlight the channelling of a fracture plan. In this stratigraphic joint studied, the conductivity variations show the complexity of a joint surface. These variations of conductivity resulted of the water flow from one channel to another one induced by the pressure variations in the drain. The last stage of this research work consists of to analyse the hydro-mechanical (H.M) behaviour of the two studied discontinuities with the UDEC 2D numerical code. The procedures of modelling examine the coupled H.M effects of a jointed rock mass affected, in a first test, by an identical joint aperture in the network, and, in a second

  17. Chaotic dynamics of red blood cells in oscillating shear flow

    NASA Astrophysics Data System (ADS)

    Bagchi, Prosenjit; Cordasco, Daniel

    2015-11-01

    A 3D computational study of deformable red blood cells in dilute suspension and subject to sinusoidally oscillating shear flow is considered. It is observed that the cell exhibits either a periodic motion or a chaotic motion. In the periodic motion, the cell reverses its orientation either about the flow direction or about the flow gradient, depending on the initial conditions. In certain parameter range, the initial conditions are forgotten and the cells become entrained in the same sequence of horizontal reversals. The chaotic dynamics is characterized by a nonperiodic sequence of horizontal and vertical reversals, and swings. The study provides the first conclusive evidence of the chaotic dynamics of fully deformable cells in oscillating flow using a deterministic numerical model without the introduction of any stochastic noise. An analysis of the chaotic dynamics shows that chaos is only possible in certain frequency bands when the cell membrane can rotate by a certain amount allowing the cells to swing near the maximum shear rate. We make a novel observation that the occurrence of the vertical or horizontal reversal depends only on whether a critical angle, that is independent of the flow frequency, is exceeded at the instant of flow reversal.

  18. Electrochemical cell for rebalancing REDOX flow system

    NASA Technical Reports Server (NTRS)

    Thaller, L. H. (Inventor)

    1979-01-01

    An electrically rechargeable REDOX cell or battery system including one of more rebalancing cells is described. Each rebalancing cell is divided into two chambers by an ion permeable membrane. The first chamber is fed with gaseous hydrogen and a cathode fluid which is circulated through the cathode chamber of the REDOX cell is also passed through the second chamber of the rebalancing cell. Electrochemical reactions take place on the surface of insert electrodes in the first and second chambers to rebalance the electrochemical capacity of the anode and cathode fluids of the REDOX system.

  19. oriGNAI3: a narrow zone of preferential replication initiation in mammalian cells identified by 2D gel and competitive PCR replicon mapping techniques.

    PubMed Central

    Toledo, F; Baron, B; Fernandez, M A; Lachagès, A M; Mayau, V; Buttin, G; Debatisse, M

    1998-01-01

    The nature of mammalian origins of DNA replication remains controversial and this is primarily because two-dimensional gel replicon mapping techniques have identified broad zones of replication initiation whereas several other techniques, such as quantitative PCR, have disclosed more discrete sites of initiation at the same chromosomal loci. In this report we analyze the replication of an amplified genomic region encompassing the 3'-end of the GNAI3 gene, the entire GNAT2 gene and the intergenic region between them in exponentially growing Chinese hamster fibroblasts. These cells express GNAI3 but not GNAT2 . The replication pattern was first analyzed by two-dimensional neutral-alkaline gel electrophoresis. Surprisingly, the results revealed a small preferential zone of replication initiation, of at most 1.7 kb, located in a limited part of the GNAI3 - GNAT2 intergenic region. Mapping of this initiation zone was then confirmed by quantitative PCR. The agreement between the two techniques exploited here strengthens the hypothesis that preferred sites of replication initiation do exist in mammalian genomes. PMID:9580680

  20. Methods for improved resolution of flow electrophoresis cells

    NASA Technical Reports Server (NTRS)

    Mccreight, L. R.; Fogal, G. L.

    1974-01-01

    First method involves remote adjusting of zeta potential. Second approach sandwiches two conducting metal plates between opposite cell walls and thin insulating layer. Third method forces buffer to flow in direction opposite particle streams.

  1. Gas-Particle Interactions in a Microgravity Flow Cell

    NASA Technical Reports Server (NTRS)

    Louge, Michel; Jenkins, James

    1999-01-01

    We are developing a microgravity flow cell in which to study the interaction of a flowing gas with relatively massive particles that collide with each other and with the moving boundaries of the cell. The absence of gravity makes possible the independent control of the relative motion of the boundaries and the flow of the gas. The cell will permit gas-particle interactions to be studied over the entire range of flow conditions over which the mixture is not turbulent. Within this range, we shall characterize the viscous dissipation of the energy of the particle fluctuations, measure the influence of particle-phase viscosity on the pressure drop along the cell, and observe the development of localized inhomogeneities that are likely to be associated with the onset of clusters. These measurements and observations should contribute to an understanding of the essential physics of pneumatic transport.

  2. Unparticle example in 2D.

    PubMed

    Georgi, Howard; Kats, Yevgeny

    2008-09-26

    We discuss what can be learned about unparticle physics by studying simple quantum field theories in one space and one time dimension. We argue that the exactly soluble 2D theory of a massless fermion coupled to a massive vector boson, the Sommerfield model, is an interesting analog of a Banks-Zaks model, approaching a free theory at high energies and a scale-invariant theory with nontrivial anomalous dimensions at low energies. We construct a toy standard model coupling to the fermions in the Sommerfield model and study how the transition from unparticle behavior at low energies to free particle behavior at high energies manifests itself in interactions with the toy standard model particles.

  3. Flow of Red Blood Cells in Stenosed Microvessels

    PubMed Central

    Vahidkhah, Koohyar; Balogh, Peter; Bagchi, Prosenjit

    2016-01-01

    A computational study is presented on the flow of deformable red blood cells in stenosed microvessels. It is observed that the Fahraeus-Lindqvist effect is significantly enhanced due to the presence of a stenosis. The apparent viscosity of blood is observed to increase by several folds when compared to non-stenosed vessels. An asymmetric distribution of the red blood cells, caused by geometric focusing in stenosed vessels, is observed to play a major role in the enhancement. The asymmetry in cell distribution also results in an asymmetry in average velocity and wall shear stress along the length of the stenosis. The discrete motion of the cells causes large time-dependent fluctuations in flow properties. The root-mean-square of flow rate fluctuations could be an order of magnitude higher than that in non-stenosed vessels. Several folds increase in Eulerian velocity fluctuation is also observed in the vicinity of the stenosis. Surprisingly, a transient flow reversal is observed upstream a stenosis but not downstream. The asymmetry and fluctuations in flow quantities and the flow reversal would not occur in absence of the cells. It is concluded that the flow physics and its physiological consequences are significantly different in micro- versus macrovascular stenosis. PMID:27319318

  4. Flow of Red Blood Cells in Stenosed Microvessels

    NASA Astrophysics Data System (ADS)

    Vahidkhah, Koohyar; Balogh, Peter; Bagchi, Prosenjit

    2016-06-01

    A computational study is presented on the flow of deformable red blood cells in stenosed microvessels. It is observed that the Fahraeus-Lindqvist effect is significantly enhanced due to the presence of a stenosis. The apparent viscosity of blood is observed to increase by several folds when compared to non-stenosed vessels. An asymmetric distribution of the red blood cells, caused by geometric focusing in stenosed vessels, is observed to play a major role in the enhancement. The asymmetry in cell distribution also results in an asymmetry in average velocity and wall shear stress along the length of the stenosis. The discrete motion of the cells causes large time-dependent fluctuations in flow properties. The root-mean-square of flow rate fluctuations could be an order of magnitude higher than that in non-stenosed vessels. Several folds increase in Eulerian velocity fluctuation is also observed in the vicinity of the stenosis. Surprisingly, a transient flow reversal is observed upstream a stenosis but not downstream. The asymmetry and fluctuations in flow quantities and the flow reversal would not occur in absence of the cells. It is concluded that the flow physics and its physiological consequences are significantly different in micro- versus macrovascular stenosis.

  5. Thermally-induced single-crystal-to-single-crystal transformations from a 2D two-fold interpenetrating square lattice layer to a 3D four-fold interpenetrating diamond framework and its application in dye-sensitized solar cells.

    PubMed

    Gao, Song; Fan, Rui Qing; Wang, Xin Ming; Wei, Li Guo; Song, Yang; Du, Xi; Xing, Kai; Wang, Ping; Yang, Yu Lin

    2016-07-28

    In this work, a rare 2D → 3D single-crystal-to-single-crystal transformation (SCSC) is observed in metal-organic coordination complexes, which is triggered by thermal treatment. The 2D two-fold interpenetrating square lattice layer [Cd(IBA)2]n (1) is irreversibly converted into a 3D four-fold interpenetrating diamond framework {[Cd(IBA)2(H2O)]·2.5H2O}n (2) (HIBA = 4-(1H-imidazol-1-yl)benzoic acid). Consideration is given to these two complexes with different interpenetrating structures and dimensionality, and their influence on photovoltaic properties are studied. Encouraged by the UV-visible absorption and HOMO-LUMO energy states matched for sensitizing TiO2, the two complexes are employed in combination with N719 in dye-sensitized solar cells (DSSCs) to compensate absorption in the ultraviolet and blue-violet region, offset competitive visible light absorption of I3(-) and reducing charge the recombination of injected electrons. After co-sensitization with 1 and 2, the device co-sensitized by 1/N719 and 2/N719 to yield overall efficiencies of 7.82% and 8.39%, which are 19.94% and 28.68% higher than that of the device sensitized only by N719 (6.52%). Consequently, high dimensional interpenetrating complexes could serve as excellent co-sensitizers and have application in DSSCs. PMID:27356177

  6. Thermally-induced single-crystal-to-single-crystal transformations from a 2D two-fold interpenetrating square lattice layer to a 3D four-fold interpenetrating diamond framework and its application in dye-sensitized solar cells.

    PubMed

    Gao, Song; Fan, Rui Qing; Wang, Xin Ming; Wei, Li Guo; Song, Yang; Du, Xi; Xing, Kai; Wang, Ping; Yang, Yu Lin

    2016-07-28

    In this work, a rare 2D → 3D single-crystal-to-single-crystal transformation (SCSC) is observed in metal-organic coordination complexes, which is triggered by thermal treatment. The 2D two-fold interpenetrating square lattice layer [Cd(IBA)2]n (1) is irreversibly converted into a 3D four-fold interpenetrating diamond framework {[Cd(IBA)2(H2O)]·2.5H2O}n (2) (HIBA = 4-(1H-imidazol-1-yl)benzoic acid). Consideration is given to these two complexes with different interpenetrating structures and dimensionality, and their influence on photovoltaic properties are studied. Encouraged by the UV-visible absorption and HOMO-LUMO energy states matched for sensitizing TiO2, the two complexes are employed in combination with N719 in dye-sensitized solar cells (DSSCs) to compensate absorption in the ultraviolet and blue-violet region, offset competitive visible light absorption of I3(-) and reducing charge the recombination of injected electrons. After co-sensitization with 1 and 2, the device co-sensitized by 1/N719 and 2/N719 to yield overall efficiencies of 7.82% and 8.39%, which are 19.94% and 28.68% higher than that of the device sensitized only by N719 (6.52%). Consequently, high dimensional interpenetrating complexes could serve as excellent co-sensitizers and have application in DSSCs.

  7. Low cost, radial flow, solid oxide fuel cell

    NASA Astrophysics Data System (ADS)

    Petrik, M. A.

    The Interscience Radial Flow (IRF) SOFC is designed to minimize problems in high-temperature operation and for low-cost fabrication. The cell has planar, non-sintered construction, uses particulate materials to form porous electrodes, and has internal radial flow. The object of this phase was to demonstrate feasibility of multi-cell stack operation. Performance milestone was 15% DC HHV efficiency with hydrogen at greater than 50 mW/sq cm over 100 h.

  8. Flow cytometric immunofluorescence of rat anterior pituitary cells

    NASA Technical Reports Server (NTRS)

    Hatfield, J. Michael; Hymer, W. C.

    1985-01-01

    A flow cytometric immunofluorescence technique was developed for the quantification of growth hormone, prolactin, and luteinizing hormone producing cells. The procedure is based on indirect-immunofluorescence of intracellular hormone using an EPICS V cell sorter and can objectively count 50,000 cells in about 3 minutes. It can be used to study the dynamics of pituitary cell populations under various physiological and pharmacological conditions.

  9. Functional characterization of CYP2D6 enhancer polymorphisms

    PubMed Central

    Wang, Danxin; Papp, Audrey C.; Sun, Xiaochun

    2015-01-01

    CYP2D6 metabolizes nearly 25% of clinically used drugs. Genetic polymorphisms cause large inter-individual variability in CYP2D6 enzyme activity and are currently used as biomarker to predict CYP2D6 metabolizer phenotype. Previously, we had identified a region 115 kb downstream of CYP2D6 as enhancer for CYP2D6, containing two completely linked single nucleotide polymorphisms (SNPs), rs133333 and rs5758550, associated with enhanced transcription. However, the enhancer effect on CYP2D6 expression, and the causative variant, remained to be ascertained. To characterize the CYP2D6 enhancer element, we applied chromatin conformation capture combined with the next-generation sequencing (4C assays) and chromatin immunoprecipitation with P300 antibody, in HepG2 and human primary culture hepatocytes. The results confirmed the role of the previously identified enhancer region in CYP2D6 expression, expanding the number of candidate variants to three highly linked SNPs (rs133333, rs5758550 and rs4822082). Among these, only rs5758550 demonstrated regulating enhancer activity in a reporter gene assay. Use of clustered regularly interspaced short palindromic repeats mediated genome editing in HepG2 cells targeting suspected enhancer regions decreased CYP2D6 mRNA expression by 70%, only upon deletion of the rs5758550 region. These results demonstrate robust effects of both the enhancer element and SNP rs5758550 on CYP2D6 expression, supporting consideration of rs5758550 for CYP2D6 genotyping panels to yield more accurate phenotype prediction. PMID:25381333

  10. Flow field measurements in the cell culture unit

    NASA Technical Reports Server (NTRS)

    Walker, Stephen; Wilder, Mike; Dimanlig, Arsenio; Jagger, Justin; Searby, Nancy

    2002-01-01

    The cell culture unit (CCU) is being designed to support cell growth for long-duration life science experiments on the International Space Station (ISS). The CCU is a perfused loop system that provides a fluid environment for controlled cell growth experiments within cell specimen chambers (CSCs), and is intended to accommodate diverse cell specimen types. Many of the functional requirements depend on the fluid flow field within the CSC (e.g., feeding and gas management). A design goal of the CCU is to match, within experimental limits, all environmental conditions, other than the effects of gravity on the cells, whether the hardware is in microgravity ( micro g), normal Earth gravity, or up to 2g on the ISS centrifuge. In order to achieve this goal, two steps are being taken. The first step is to characterize the environmental conditions of current 1g cell biology experiments being performed in laboratories using ground-based hardware. The second step is to ensure that the design of the CCU allows the fluid flow conditions found in 1g to be replicated from microgravity up to 2g. The techniques that are being used to take these steps include flow visualization, particle image velocimetry (PIV), and computational fluid dynamics (CFD). Flow visualization using the injection of dye has been used to gain a global perspective of the characteristics of the CSC flow field. To characterize laboratory cell culture conditions, PIV is being used to determine the flow field parameters of cell suspension cultures grown in Erlenmeyer flasks on orbital shakers. These measured parameters will be compared to PIV measurements in the CSCs to ensure that the flow field that cells encounter in CSCs is within the bounds determined for typical laboratory experiments. Using CFD, a detailed simulation is being developed to predict the flow field within the CSC for a wide variety of flow conditions, including microgravity environments. Results from all these measurements and analyses of the

  11. Secretory pathways generating immunosuppressive NKG2D ligands

    PubMed Central

    Baragaño Raneros, Aroa; Suarez-Álvarez, Beatriz; López-Larrea, Carlos

    2014-01-01

    Natural Killer Group 2 member D (NKG2D) activating receptor, present on the surface of various immune cells, plays an important role in activating the anticancer immune response by their interaction with stress-inducible NKG2D ligands (NKG2DL) on transformed cells. However, cancer cells have developed numerous mechanisms to evade the immune system via the downregulation of NKG2DL from the cell surface, including the release of NKG2DL from the cell surface in a soluble form. Here, we review the mechanisms involved in the production of soluble NKG2DL (sNKG2DL) and the potential therapeutic strategies aiming to block the release of these immunosuppressive ligands. Therapeutically enabling the NKG2D-NKG2DL interaction would promote immunorecognition of malignant cells, thus abrogating disease progression. PMID:25050215

  12. An innovative hybrid 3D analytic-numerical model for air breathing parallel channel counter-flow PEM fuel cells.

    PubMed

    Tavčar, Gregor; Katrašnik, Tomaž

    2014-01-01

    The parallel straight channel PEM fuel cell model presented in this paper extends the innovative hybrid 3D analytic-numerical (HAN) approach previously published by the authors with capabilities to address ternary diffusion systems and counter-flow configurations. The model's core principle is modelling species transport by obtaining a 2D analytic solution for species concentration distribution in the plane perpendicular to the cannel gas-flow and coupling consecutive 2D solutions by means of a 1D numerical pipe-flow model. Electrochemical and other nonlinear phenomena are coupled to the species transport by a routine that uses derivative approximation with prediction-iteration. The latter is also the core of the counter-flow computation algorithm. A HAN model of a laboratory test fuel cell is presented and evaluated against a professional 3D CFD simulation tool showing very good agreement between results of the presented model and those of the CFD simulation. Furthermore, high accuracy results are achieved at moderate computational times, which is owed to the semi-analytic nature and to the efficient computational coupling of electrochemical kinetics and species transport.

  13. An innovative hybrid 3D analytic-numerical model for air breathing parallel channel counter-flow PEM fuel cells.

    PubMed

    Tavčar, Gregor; Katrašnik, Tomaž

    2014-01-01

    The parallel straight channel PEM fuel cell model presented in this paper extends the innovative hybrid 3D analytic-numerical (HAN) approach previously published by the authors with capabilities to address ternary diffusion systems and counter-flow configurations. The model's core principle is modelling species transport by obtaining a 2D analytic solution for species concentration distribution in the plane perpendicular to the cannel gas-flow and coupling consecutive 2D solutions by means of a 1D numerical pipe-flow model. Electrochemical and other nonlinear phenomena are coupled to the species transport by a routine that uses derivative approximation with prediction-iteration. The latter is also the core of the counter-flow computation algorithm. A HAN model of a laboratory test fuel cell is presented and evaluated against a professional 3D CFD simulation tool showing very good agreement between results of the presented model and those of the CFD simulation. Furthermore, high accuracy results are achieved at moderate computational times, which is owed to the semi-analytic nature and to the efficient computational coupling of electrochemical kinetics and species transport. PMID:25125112

  14. Canard configured aircraft with 2-D nozzle

    NASA Technical Reports Server (NTRS)

    Child, R. D.; Henderson, W. P.

    1978-01-01

    A closely-coupled canard fighter with vectorable two-dimensional nozzle was designed for enhanced transonic maneuvering. The HiMAT maneuver goal of a sustained 8g turn at a free-stream Mach number of 0.9 and 30,000 feet was the primary design consideration. The aerodynamic design process was initiated with a linear theory optimization minimizing the zero percent suction drag including jet effects and refined with three-dimensional nonlinear potential flow techniques. Allowances were made for mutual interference and viscous effects. The design process to arrive at the resultant configuration is described, and the design of a powered 2-D nozzle model to be tested in the LRC 16-foot Propulsion Wind Tunnel is shown.

  15. Computational cell analysis for label-free detection of cell properties in a microfluidic laminar flow.

    PubMed

    Zhang, Alex Ce; Gu, Yi; Han, Yuanyuan; Mei, Zhe; Chiu, Yu-Jui; Geng, Lina; Cho, Sung Hwan; Lo, Yu-Hwa

    2016-06-20

    Although a flow cytometer, being one of the most popular research and clinical tools for biomedicine, can analyze cells based on the cell size, internal structures such as granularity, and molecular markers, it provides little information about the physical properties of cells such as cell stiffness and physical interactions between the cell membrane and fluid. In this paper, we propose a computational cell analysis technique using cells' different equilibrium positions in a laminar flow. This method utilizes a spatial coding technique to acquire the spatial position of the cell in a microfluidic channel and then uses mathematical algorithms to calculate the ratio of cell mixtures. Most uniquely, the invented computational cell analysis technique can unequivocally detect the subpopulation of each cell type without labeling even when the cell type shows a substantial overlap in the distribution plot with other cell types, a scenario limiting the use of conventional flow cytometers and machine learning techniques. To prove this concept, we have applied the computation method to distinguish live and fixed cancer cells without labeling, count neutrophils from human blood, and distinguish drug treated cells from untreated cells. Our work paves the way for using computation algorithms and fluidic dynamic properties for cell classification, a label-free method that can potentially classify over 200 types of human cells. Being a highly cost-effective cell analysis method complementary to flow cytometers, our method can offer orthogonal tests in companion with flow cytometers to provide crucial information for biomedical samples. PMID:27163941

  16. Upward swimming of a sperm cell in shear flow

    NASA Astrophysics Data System (ADS)

    Omori, Toshihiro; Ishikawa, Takuji

    2016-03-01

    Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation.

  17. Upward swimming of a sperm cell in shear flow.

    PubMed

    Omori, Toshihiro; Ishikawa, Takuji

    2016-03-01

    Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation. PMID:27078385

  18. Upward swimming of a sperm cell in shear flow.

    PubMed

    Omori, Toshihiro; Ishikawa, Takuji

    2016-03-01

    Mammalian sperm cells are required to swim over long distances, typically around 1000-fold their own length. They must orient themselves and maintain a swimming motion to reach the ovum, or egg cell. Although the mechanism of long-distance navigation is still unclear, one possible mechanism, rheotaxis, was reported recently. This work investigates the mechanism of the rheotaxis in detail by simulating the motions of a sperm cell in shear flow adjacent to a flat surface. A phase diagram was developed to show the sperm's swimming motion under different shear rates, and for varying flagellum waveform conditions. The results showed that, under shear flow, the sperm is able to hydrodynamically change its swimming direction, allowing it to swim upwards against the flow, which suggests that the upward swimming of sperm cells can be explained using fluid mechanics, and this can then be used to further understand physiology of sperm cell navigation.

  19. Flow distribution in the manifold of PEM fuel cell stack

    NASA Astrophysics Data System (ADS)

    Chen, Chung-Hsien; Jung, Shiauh-Ping; Yen, Shi-Chern

    In this study, the pressure variation and the flow distribution in the manifold of a fuel-cell stack are simulated by a computational fluid dynamics (CFD) approach. Two dimensional stack model composed of 72 cells filled with porous media is constructed to evaluate pressure drop caused by channel flow resistance. In order to simplify this model, electrochemical reactions, heat and mass transport phenomena are ignored and air is treated as working fluid to investigate flow distribution in stacks. Design parameters such as the permeability of the porous media, the manifold width and the air feeding rate were changed to estimate uniformity of the flow distribution in the manifold. A momentum-balance theory and a pressure-drop model are presented to explain the physical mechanism of flow distribution. Modeling results indicate that both the channel resistance and the manifold width can enhance the uniformity of the flow distribution. In addition, a lower air feeding rate can also enhance the uniformity of flow distribution. However, excessive pressure drop is not beneficial for realistic applications of a fuel-cell stack and hence enhanced manifold width is a better solution for flow distribution.

  20. Expression of toll-like receptors by human muscle cells in vitro and in vivo: TLR3 is highly expressed in inflammatory and HIV myopathies, mediates IL-8 release and up-regulation of NKG2D-ligands.

    PubMed

    Schreiner, Bettina; Voss, Joachim; Wischhusen, Jörg; Dombrowski, Yvonne; Steinle, Alexander; Lochmüller, Hanns; Dalakas, Marinos; Melms, Arthur; Wiendl, Heinz

    2006-01-01

    The particular microenvironment of the skeletal muscle can be the site of complex immune reactions. Toll-like receptors (TLRs) mediate inflammatory stimuli from pathogens and endogenous danger signals and link the innate and adaptive immune system. We investigated innate immune responses in human muscle. Analyzing TLR1-9 mRNA in cultured myoblasts and rhabdomyosarcoma cells, we found constitutive expression of TLR3. The TLR3 ligand Poly (I:C), a synthetic analog of dsRNA, and IFN-gamma increased TLR3 levels. TLR3 was mainly localized intracellularly and regulated at the protein level. Poly (I:C) challenge 1) activated nuclear factor-kappaB (NF-kappaB), 2) increased IL-8 release, and 3) up-regulated NKG2D ligands and NK-cell-mediated lysis of muscle cells. We examined muscle biopsy specimens of 6 HIV patients with inclusion body myositis/polymyositis (IBM/PM), 7 cases of sporadic IBM and 9 nonmyopathic controls for TLR3 expression. TLR3 mRNA levels were elevated in biopsy specimens from patients with IBM and HIV-myopathies. Muscle fibers in inflammatory myopathies expressed TLR3 in close proximity of infiltrating mononuclear cells. Taken together, our study suggests an important role of TLR3 in the immunobiology of muscle, and has substantial implications for the understanding of the pathogenesis of inflammatory myopathies or therapeutic interventions like vaccinations or gene transfer.

  1. Rapid Cell Population Identification in Flow Cytometry Data*

    PubMed Central

    Aghaeepour, Nima; Nikolic, Radina; Hoos, Holger H.; Brinkman, Ryan R.

    2011-01-01

    We have developed flowMeans, a time-efficient and accurate method for automated identification of cell populations in flow cytometry (FCM) data based on K-means clustering. Unlike traditional K-means, flowMeans can identify concave cell populations by modelling a single population with multiple clusters. flowMeans uses a change point detection algorithm to determine the number of sub-populations, enabling the method to be used in high throughput FCM data analysis pipelines. Our approach compares favourably to manual analysis by human experts and current state-of-the-art automated gating algorithms. flowMeans is freely available as an open source R package through Bioconductor. PMID:21182178

  2. Fluid flow plate for decreased density of fuel cell assembly

    DOEpatents

    Vitale, Nicholas G.

    1999-01-01

    A fluid flow plate includes first and second outward faces. Each of the outward faces has a flow channel thereon for carrying respective fluid. At least one of the fluids serves as reactant fluid for a fuel cell of a fuel cell assembly. One or more pockets are formed between the first and second outward faces for decreasing density of the fluid flow plate. A given flow channel can include one or more end sections and an intermediate section. An interposed member can be positioned between the outward faces at an interface between an intermediate section, of one of the outward faces, and an end section, of that outward face. The interposed member can serve to isolate the reactant fluid from the opposing outward face. The intermediate section(s) of flow channel(s) on an outward face are preferably formed as a folded expanse.

  3. Combustor air flow control method for fuel cell apparatus

    DOEpatents

    Clingerman, Bruce J.; Mowery, Kenneth D.; Ripley, Eugene V.

    2001-01-01

    A method for controlling the heat output of a combustor in a fuel cell apparatus to a fuel processor where the combustor has dual air inlet streams including atmospheric air and fuel cell cathode effluent containing oxygen depleted air. In all operating modes, an enthalpy balance is provided by regulating the quantity of the air flow stream to the combustor to support fuel cell processor heat requirements. A control provides a quick fast forward change in an air valve orifice cross section in response to a calculated predetermined air flow, the molar constituents of the air stream to the combustor, the pressure drop across the air valve, and a look up table of the orifice cross sectional area and valve steps. A feedback loop fine tunes any error between the measured air flow to the combustor and the predetermined air flow.

  4. Fuel cell with metal screen flow-field

    DOEpatents

    Wilson, M.S.; Zawodzinski, C.

    1998-08-25

    A polymer electrolyte membrane (PEM) fuel cell is provided with electrodes supplied with a reactant on each side of a catalyzed membrane assembly (CMA). The fuel cell includes a metal mesh defining a rectangular flow-field pattern having an inlet at a first corner and an outlet at a second corner located on a diagonal from the first corner, wherein all flow paths from the inlet to the outlet through the square flow field pattern are equivalent to uniformly distribute the reactant over the CMA. In a preferred form of metal mesh, a square weave screen forms the flow-field pattern. In a particular characterization of the present invention, a bipolar plate electrically connects adjacent fuel cells, where the bipolar plate includes a thin metal foil having an anode side and a cathode side; a first metal mesh on the anode side of the thin metal foil; and a second metal mesh on the cathode side of the thin metal foil. In another characterization of the present invention, a cooling plate assembly cools adjacent fuel cells, where the cooling plate assembly includes an anode electrode and a cathode electrode formed of thin conducting foils; and a metal mesh flow field there between for distributing cooling water flow over the electrodes to remove heat generated by the fuel cells. 11 figs.

  5. Fuel cell with metal screen flow-field

    DOEpatents

    Wilson, Mahlon S.; Zawodzinski, Christine

    2001-01-01

    A polymer electrolyte membrane (PEM) fuel cell is provided with electrodes supplied with a reactant on each side of a catalyzed membrane assembly (CMA). The fuel cell includes a metal mesh defining a rectangular flow-field pattern having an inlet at a first corner and an outlet at a second corner located on a diagonal from the first corner, wherein all flow paths from the inlet to the outlet through the square flow field pattern are equivalent to uniformly distribute the reactant over the CMA. In a preferred form of metal mesh, a square weave screen forms the flow-field pattern. In a particular characterization of the present invention, a bipolar plate electrically connects adjacent fuel cells, where the bipolar plate includes a thin metal foil having an anode side and a cathode side; a first metal mesh on the anode side of the thin metal foil; and a second metal mesh on the cathode side of the thin metal foil. In another characterization of the present invention, a cooling plate assembly cools adjacent fuel cells, where the cooling plate assembly includes an anode electrode and a cathode electrode formed of thin conducting foils; and a metal mesh flow field therebetween for distributing cooling water flow over the electrodes to remove heat generated by the fuel cells.

  6. Fuel cell with metal screen flow-field

    DOEpatents

    Wilson, Mahlon S.; Zawodzinski, Christine

    1998-01-01

    A polymer electrolyte membrane (PEM) fuel cell is provided with electrodes supplied with a reactant on each side of a catalyzed membrane assembly (CMA). The fuel cell includes a metal mesh defining a rectangular flow-field pattern having an inlet at a first corner and an outlet at a second corner located on a diagonal from the first corner, wherein all flow paths from the inlet to the outlet through the square flow field pattern are equivalent to uniformly distribute the reactant over the CMA. In a preferred form of metal mesh, a square weave screen forms the flow-field pattern. In a particular characterization of the present invention, a bipolar plate electrically connects adjacent fuel cells, where the bipolar plate includes a thin metal foil having an anode side and a cathode side; a first metal mesh on the anode side of the thin metal foil; and a second metal mesh on the cathode side of the thin metal foil. In another characterization of the present invention, a cooling plate assembly cools adjacent fuel cells, where the cooling plate assembly includes an anode electrode and a cathode electrode formed of thin conducting foils; and a metal mesh flow field therebetween for distributing cooling water flow over the electrodes to remove heat generated by the fuel cells.

  7. Inertia-dependent dynamics of three-dimensional vesicles and red blood cells in shear flow.

    PubMed

    Luo, Zheng Yuan; Wang, Shu Qi; He, Long; Xu, Feng; Bai, Bo Feng

    2013-10-28

    A three-dimensional (3D) simulation study of the effect of inertia on the dynamics of vesicles and red blood cells (RBCs) has not been reported. Here, we developed a 3D model based on the front tracking method to investigate how inertia affects the dynamics of spherical/non-spherical vesicles and biconcave-shaped RBCs with the Reynolds number ranging from 0.1 to 10. The results showed that inertia induced non-spherical vesicles transitioned from tumbling to swinging, which was not observed in previous 2D models. The critical viscosity ratio of inner/outer fluids for the tumbling–swinging transition remarkably increased with an increasing Reynolds number. The deformation of vesicles was greatly enhanced by inertia, and the frequency of tumbling and tank-treading was significantly decreased by inertia. We also found that RBCs can transit from tumbling to steady tank-treading through the swinging regime when the Reynolds number increased from 0.1 to 10. These results indicate that inertia needs to be considered at moderate Reynolds number (Re ~ 1) in the study of blood flow in the human body and the flow of deformable particle suspension in inertial microfluidic devices. The developed 3D model provided new insights into the dynamics of RBCs under shear flow, thus holding great potential to better understand blood flow behaviors under normal/disease conditions.

  8. New all-vanadium redox flow cell

    NASA Astrophysics Data System (ADS)

    Skyllas-Kazacos, M.; Rychcik, M.; Robins, R. G.; Fane, A. G.; Green, M. A.

    1986-05-01

    A laboratory-scale cell was constructed to test the performance of V(II)/V(III) and V(IV)/V(V) half-cells in an all-vanadium redox battery. Graphite plates were used as electrodes, and the membrane was manufactured from a sulfonated polyehylene anion-selective material. The average charging efficiency of the cell was over 90 percent. Stability tests on the reduced and oxidized electrolytes, measured over the temperature range of -5 C to 60 C, showed no accelerated decomposition at high temperatures and no crystallization at the lower temperatures. After prolonged usage, however, a slow deterioration of the positive electrode and the membrane was observed.

  9. Full dynamics of a red blood cell in shear flow.

    PubMed

    Dupire, Jules; Socol, Marius; Viallat, Annie

    2012-12-18

    At the cellular scale, blood fluidity and mass transport depend on the dynamics of red blood cells in blood flow, specifically on their deformation and orientation. These dynamics are governed by cellular rheological properties, such as internal viscosity and cytoskeleton elasticity. In diseases in which cell rheology is altered genetically or by parasitic invasion or by changes in the microenvironment, blood flow may be severely impaired. The nonlinear interplay between cell rheology and flow may generate complex dynamics, which remain largely unexplored experimentally. Under simple shear flow, only two motions, "tumbling" and "tank-treading," have been described experimentally and relate to cell mechanics. Here, we elucidate the full dynamics of red blood cells in shear flow by coupling two videomicroscopy approaches providing multidirectional pictures of cells, and we analyze the mechanical origin of the observed dynamics. We show that contrary to common belief, when red blood cells flip into the flow, their orientation is determined by the shear rate. We discuss the "rolling" motion, similar to a rolling wheel. This motion, which permits the cells to avoid energetically costly deformations, is a true signature of the cytoskeleton elasticity. We highlight a hysteresis cycle and two transient dynamics driven by the shear rate: an intermittent regime during the "tank-treading-to-flipping" transition and a Frisbee-like "spinning" regime during the "rolling-to-tank-treading" transition. Finally, we reveal that the biconcave red cell shape is highly stable under moderate shear stresses, and we interpret this result in terms of stress-free shape and elastic buckling. PMID:23213229

  10. Wavelet characterization of 2D turbulence and intermittency in magnetized electron plasmas

    NASA Astrophysics Data System (ADS)

    Romé, M.; Chen, S.; Maero, G.

    2016-06-01

    A study of the free relaxation of turbulence in a two-dimensional (2D) flow is presented, with a focus on the role of the initial vorticity conditions. Exploiting a well-known analogy with 2D inviscid incompressible fluids, the system investigated here is a magnetized pure electron plasma. The dynamics of this system are simulated by means of a 2D particle-in-cell code, starting from different spiral density (vorticity) distributions. A wavelet multiresolution analysis is adopted, which allows the coherent and incoherent parts of the flow to be separated. Comparison of the turbulent evolution in the different cases is based on the investigation of the time evolution of statistical properties, including the probability distribution functions and structure functions of the vorticity increments. It is also based on an analysis of the enstrophy evolution and its spectrum for the two components. In particular, while the statistical features assess the degree of flow intermittency, spectral analysis allows us not only to estimate the time required to reach a state of fully developed turbulence, but also estimate its dependence on the thickness of the initial spiral density distribution, accurately tracking the dynamics of both the coherent structures and the turbulent background. The results are compared with those relevant to annular initial vorticity distributions (Chen et al 2015 J. Plasma Phys. 81 495810511).

  11. Pockels-effect cell for gas-flow simulation

    NASA Technical Reports Server (NTRS)

    Weimer, D.

    1982-01-01

    A Pockels effect cell using a 75 cu cm DK*P crystal was developed and used as a gas flow simulator. Index of refraction gradients were produced in the cell by the fringing fields of parallel plate electrodes. Calibration curves for the device were obtained for index of refraction gradients in excess of .00025 m.

  12. Segmentation of touching cell nuclei using gradient flow tracking.

    PubMed

    Li, G; Liu, T; Nie, J; Guo, L; Chen, J; Zhu, J; Xia, W; Mara, A; Holley, S; Wong, S T C

    2008-07-01

    Reliable cell nuclei segmentation is an important yet unresolved problem in biological imaging studies. This paper presents a novel computerized method for robust cell nuclei segmentation based on gradient flow tracking. This method is composed of three key steps: (1) generate a diffused gradient vector flow field; (2) perform a gradient flow tracking procedure to attract points to the basin of a sink; and (3) separate the image into small regions, each containing one nucleus and nearby peripheral background, and perform local adaptive thresholding in each small region to extract the cell nucleus from the background. To show the generality of the proposed method, we report the validation and experimental results using microscopic image data sets from three research labs, with both over-segmentation and under-segmentation rates below 3%. In particular, this method is able to segment closely juxtaposed or clustered cell nuclei, with high sensitivity and specificity in different situations.

  13. Margination of White Blood Cells in Microcapillary Flow

    NASA Astrophysics Data System (ADS)

    Fedosov, Dmitry A.; Fornleitner, Julia; Gompper, Gerhard

    2012-01-01

    Margination of white blood cells (WBCs) towards vessel walls is an essential precondition for their efficient adhesion to the vascular endothelium. We perform numerical simulations with a two-dimensional blood flow model to investigate the dependence of WBC margination on hydrodynamic interactions of blood cells with the vessel walls, as well as on their collective behavior and deformability. We find WBC margination to be optimal in intermediate ranges of red blood cell (RBC) volume fractions and flow rates, while, beyond these ranges, it is substantially attenuated. RBC aggregation enhances WBC margination, while WBC deformability reduces it. These results are combined in state diagrams, which identify WBC margination for a wide range of flow and cell suspension conditions.

  14. Proteomic analysis of human bladder epithelial cells by 2D blue native SDS-PAGE reveals TCDD-induced alterations of calcium and iron homeostasis possibly mediated by nitric oxide.

    PubMed

    Verma, Nisha; Pink, Mario; Petrat, Frank; Rettenmeier, Albert W; Schmitz-Spanke, Simone

    2015-01-01

    A proteomic analysis of the interaction among multiprotein complexes involved in 2,3,7,8-dibenzo-p-dioxin (TCDD)-mediated toxicity in urinary bladder epithelial RT4 cells was performed using two-dimensional blue native SDS-PAGE (2D BN/SDS-PAGE). To enrich the protein complexes, unexposed and TCDD-exposed cells were fractionated. BN/SDS-PAGE of the resulting fractions led to an effective separation of proteins and protein complexes of various origins, including cell membrane, mitochondria, and other intracellular compartments. Major differences between the proteome of control and exposed cells involved the alteration of many calcium-regulated proteins (calmodulin, protein S100-A2, annexin A5, annexin A10, gelsolin isoform b) and iron-regulated proteins (ferritin, heme-binding protein 2, transferrin). On the basis of these findings, the intracellular calcium concentration was determined, revealing a significant increase after 24 h of exposure to TCDD. Moreover, the concentration of the labile iron pool (LIP) was also significantly elevated in TCDD-exposed cells. This increase was strongly inhibited by the calmodulin (CaM) antagonist W-7, which pointed toward a possible interaction between iron and calcium signaling. Because nitric oxide (NO) production was significantly enhanced in TCDD-exposed cells and was also inhibited by W-7, we hypothesize that alterations in calcium and iron homeostasis upon exposure to TCDD may be linked through NO generated by CaM-activated nitric oxide synthase. In our model, we propose that NO produced upon TCDD exposure interacts with the iron centers of iron-regulatory proteins (IRPs) that modulate the alteration of ferritin and transferrin, resulting in an augmented cellular LIP and, hence, increased toxicity.

  15. Numerical simulations of capsules and red blood cells under flow in complex geometries at non-zero Reynolds numbers

    NASA Astrophysics Data System (ADS)

    Mendez, Simon; Gibaud, Etienne; Siguenza, Julien; Nicoud, Franck; Acsiom-I3M Team

    2013-11-01

    Numerical simulation of flows of vesicles, capsules and cells is a growing field (Misbah 2012). With the objective of understanding the complex fluid-structure interactions involved in such flows, studying microcirculation and suspension rheology or improving drug vectorization, numerous research groups have developed numerical methods to compute the dynamics of deformable objects like capsules and red blood cells, composed by a drop of liquid enclosed by a membrane. However, the most mature methods rely on boundary integrals, the use of which is allowed by the Stokes flow hypothesis: boundary integral method (BIM) is thus an efficient tool to study microfluidics and microcirculation. In some flows, in particular in some medical devices, the Reynolds number may be high, which precludes the use of the BIM. In this talk, we will show how the immersed boundary method can be implemented in an unstructured finite-volume solver to tackle such flows of deformable objects. The method will be detailed and specific attention will be devoted to the validation of the solver, in particular in 2D, where reference results are scarce. Finally, applications of the method to flows of isolated cells will be shown.

  16. Liquid chromatography/Fourier transform IR spectrometry interface flow cell

    DOEpatents

    Johnson, Charles C.; Taylor, Larry T.

    1986-01-01

    A zero dead volume (ZDV) microbore high performance liquid chromatography (.mu.HPLC)/Fourier transform infrared (FTIR) interface flow cell includes an IR transparent crystal having a small diameter bore therein through which a sample liquid is passed. The interface flow cell further includes a metal holder in