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At least 865 records · Page 48

Irradiated HMEC from A-T Heterozygous Breast Tissue

Women who are heterozygous for ataxia-telangiectasia (A-T) carry a single defective ATM gene in chromosome 11 q22-23, and have been statistically determined with high significance within a defined database to be approximately 5-fold more susceptible for developing breast cancer than their noma1 counterpart. Breast cancer susceptibility of these A-T heterozygotes has been hypothesized to include consequence of response to damage caused by low levels of ionizing radiation. Prophylactic mastectomy specimens were donated by a 41 year-old obligate A-T heterozygote who was located prior to her elective surgery through an existing pedigree. Harvest of that breast tissue provided an isolate of long-term growth human mammary epithelial cells (HMEC), designated WH612/3. An isolate of presumed normal long-term growth HMEC, designated 48R, was obtained from Dr. Martha Stampfer (Lawrence Berkeley Laboratory, University of California), and the A-T heterozygous HMEC were transformed with E6 and E7 oncogenes of human papilloma virus Type-16 in the laboratory of Dr. Ray White (Hunt- Cancer Institute, University of Utah) for use in this study. The objective of this study is to study the expression of end points that may bear on cancer outcome following irradiation of HMEC. Specific end points are cell survival, cell cycle, p53 expression, and apoptosis. Survival curves, immunostaining, and flow cytometery are used to examine these end points. Radiation-induced cell killing shows less shoulder development in the survival curve for WH61U3 compared to 48R HMEC, suggesting less repair of damage in the former HMEC. Additional information is included in the original extended abstract.

Richmond, Robert↗

CD47 blockade enhances therapeutic efficacy of cisplatin against lung carcinoma in a murine model

Cisplatin (CDDP) is the first generation of platinum-based drug and is widely used to treat many cancers due to its potency. The present study aims to explore the effects of CDDP on lung carcinoma and its relationship with macrophage phagocytosis. In in vitro study, murine and human lung cancer cell lines were applied and treated with CDDP, CD47 antibody (aCD47), or CDDP plus aCD47. In in vivo study, a tumor xenograft animal model was treated with CDDP, aCD47, or CDDP plus aCD47. Real-time PCR was applied to determine the mRNA expressions. Enzyme-linked immunosorbent assay (ELISA), Western blotting, and Immunofluorescent staining were applied to determine the protein expressions. Flow cytometry was applied to analyze cell apoptosis, phagocytosis, and specific cell populations. CDDP enhanced the expressions of CD47 in lung cancer cells. Interestingly, the blockage of CD47 enhanced the macrophages' phagocytic activity on the CDDP-treated tumor cells. The treatment of CDDP and aCD47 exhibited anti-tumor effects and prolonged the LLC tumor-bearing mice survival time. Mechanistic studies revealed that the treatment of CDDP and aCD47 regulated the phagocytic activity of macrophage, percentage of CD8{sup +} T cells, and cytokines (tumor growth factor (TGF)-β, interleukin (IL)12p70, and interferon (IFN)-γ) in the tumor-bearing model. CD47 blockade enhanced therapeutic efficacy of cisplatin against lung carcinoma in vivo and in vitro.

60 APPLIED LIFE SCIENCES↗

A numerical procedure for analysis of finite rate reacting flows

Combustion processes in rocket propulsion systems are characterized by the existence of multiple, vastly differing time and length scales, as well as flow-speeds at wide variation of Mach numbers. The chemical kinetics processes in the highly active reaction zone are characterized by much smaller scales compared to fluid convective and diffusive time scales. An operator splitting procedure for transient finite rate chemistry problems has been developed using a pressure based method, which can be applied to all speed flows without difficulties. The splitting of chemical kinetics terms formed the fluid-mechanical terms of the species equation ameliorated the difficulties associated with the disparate time scales and stiffness in the set of equations which describes highly exothermic combustion. A combined efficient ordinary differential equations (ODE) solver was used to integrate the effective chemical source terms over the residence time at each grid cell. One and two dimensional reacting flow situations were carried out to demonstrate and verify the current procedure. Different chemical kinetics with different degrees of nonlinearity have also been incorporated to test the robustness and generality of the proposed method.

Shang, H. M.↗

Progress with multigrid schemes for hypersonic flow problems

Several multigrid schemes are considered for the numerical computation of viscous hypersonic flows. For each scheme, the basic solution algorithm uses upwind spatial discretization with explicit multistage time stepping. Two level versions of the various multigrid algorithms are applied to the two dimensional advection equation, and Fourier analysis is used to determine their damping properties. The capabilities of the multigrid methods are assessed by solving three different hypersonic flow problems. Some new multigrid schemes based on semicoarsening strategies are shown to be quite effective in relieving the stiffness caused by the high aspect ratio cells required to resolve high Reynolds number flows. These schemes exhibit good convergence rates for Reynolds numbers up to 200 x 10(exp 6) and Mach numbers up to 25.

Radespiel, R.↗

Physical bioenergetics: Energy fluxes, budgets, and constraints in cells

Cells are the basic units of all living matter which harness the flow of energy to drive the processes of life. While the biochemical networks involved in energy transduction are well-characterized, the energetic costs and constraints for specific cellular processes remain largely unknown. In particular, what are the energy budgets of cells? What are the constraints and limits energy flows impose on cellular processes? Do cells operate near these limits, and if so how do energetic constraints impact cellular functions? Physics has provided many tools to study nonequilibrium systems and to define physical limits, but applying these tools to cell biology remains a challenge. Physical bioenergetics, which resides at the interface of nonequilibrium physics, energy metabolism, and cell biology, seeks to understand how much energy cells are using, how they partition this energy between different cellular processes, and the associated energetic constraints. In this work, we review recent advances and discuss open questions and challenges in physical bioenergetics.

59 BASIC BIOLOGICAL SCIENCES↗

Oridonin interrupts cellular bioenergetics to suppress glioma cell growth by down‐regulating PCK2

We aim to evaluate the tumor metabolic suppressive activity of Oridonin (extract of Rabdosia rubescens ) in glioma and elucidate its potential mechanism. Effects of Oridonin on U251/U87 cells were determined by CCK8, RTCA, colony formation, flow cytometry, wound healing, and Transwell assay. Xenograft tumor model to evaluate the effect of Oridonin on glioma cells in vivo. Cellular bioenergetics were measured by Seahorse. RNA‐seq was performed to screen potential biological pathways in Oridonin treated cells. Bioinformatics analysis of PCK2 in glioma was performed based on TCGA/CGGA. Endogenous PCK2 was knocked‐down by lentivirus packaged shRNA. We found Oridonin significantly inhibited cell growth in U251/U87 in vitro and in vivo. Both oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) were decreased in Oridonin‐treated U251/U87 cells. Oridonin treatment led to PCK2 down‐regulation. Additionally, PCK2 was up‐regulated in higher grade glioma and correlated with poor outcomes. Furthermore, PCK2 depletion significantly inhibited cell growth and decreased OCR/ECAR in U251/U87 which coincided with the effects of Oridonin. Therefore, we evaluated the potent anti‐tumor property of Oridonin in glioma. Importantly, we demonstrated that PCK2 might be a novel target of Oridonin on glioma by inducing energy crisis and increasing oxidative stress.

Lin, Jianhu↗

Numerical design considerations for vapor transport deposition of metal-halide perovskite thin films

While metal-halide perovskites (MHPs) offer high efficiency and potential application in single junction and tandem solar cells, challenges remain in translating solution-based methods to commercial-scale production. Vapor transport deposition (VTD) offers advantages such as precise control over film composition and purity, as well as the possibility of achieving high thin film deposition rates for scale-up. In this work, a numerical model is developed to simulate the deposition process, analyzing the impact of parameters including source material temperature, carrier gas and dilution gas flow rates, and chamber pressure to optimize deposition rates and film uniformity. Geometrical parameters, such as throw distance, are also found to significantly affect rate and uniformity. Furthermore, the modeling results presented here offer valuable insight for improving the scalability of VTD for the processing of metal-halide perovskites.

Chemical vapor deposition↗

Stress-induced alterations in interferon production and class II histocompatibility antigen expression

Mild electric foot-shock has been shown to be a stressor that can alter immune responses. Male Lewis rats were exposed to one session of 16 5.0-s 1.6-mA foot-shocks. Production of interferon-gamma by splenocytes in response to concanavalin-A was decreased in spleens from the shocked rats compared to control spleens. Spleen cells from rats treated with nadolol, a peripherally acting beta-adrenergic receptor antagonist, and then shocked, showed dose-dependent attenuation of the suppression of interferon-gamma production. This suggests that catecholamines mediate shock-induced suppression of interferon-gamma production. The percentage of splenic mononuclear cells expressing class II histocompatibility (Ia) antigens on their surfaces from spleens of shocked rats was determined by flow cytometry. Significantly decreased class II positive mononuclear cells were present in the spleens of shocked rats in comparison to the spleens of control rats. This may reflect an alteration of cell trafficking or decreased production of class II antigens.

Non-NASA Center↗

Flux Transport and the Sun's Global Magnetic Field

The Sun s global magnetic field is produced and evolved through the emergence of magnetic flux in active regions and its transport across the solar surface by the axisymmetric differential rotation and meridional flow and the non-axisymmetric convective flows of granulation, supergranulation, and giant cell convection. Maps of the global magnetic field serve as the inner boundary condition for space weather. The photospheric magnetic field and its evolution determine the coronal and solar wind structures through which CMEs must propagate and in which solar energetic particles are accelerated and propagate. Producing magnetic maps which best represent the actual field configuration at any instant requires knowing the magnetic field over the observed hemisphere as well as knowing the flows that transport flux. From our Earth-based vantage point we only observe the front-side hemisphere and each pole is observable for only six months of the year at best. Models for the surface magnetic flux transport can be used to provide updates to the magnetic field configuration in those unseen regions. In this presentation I will describe successes and failures of surface flux transport and present new observations on the structure, the solar cycle variability, and the evolution of the flows involved in magnetic flux transport. I find that supergranules play the dominant role due to their strong flow velocities and long lifetimes. Flux is transported by differential rotation and meridional flow only to the extent that the supergranules participate in those two flows.

Hathaway, David H.↗

Tetrahedral-Mesh Simulation of Turbulent Flows with the Space-Time Conservative Schemes

Direct numerical simulations of turbulent flows are predominantly carried out using structured, hexahedral meshes despite decades of development in unstructured mesh methods. Tetrahedral meshes offer ease of mesh generation around complex geometries and the potential of an orientation free grid that would provide un-biased small-scale dissipation and more accurate intermediate scale solutions. However, due to the lack of consistent multi-dimensional numerical formulations in conventional schemes for triangular and tetrahedral meshes at the cell interfaces, numerical issues exist when flow discontinuities or stagnation regions are present. The space-time conservative conservation element solution element (CESE) method - due to its Riemann-solver-free shock capturing capabilities, non-dissipative baseline schemes, and flux conservation in time as well as space - has the potential to more accurately simulate turbulent flows using unstructured tetrahedral meshes. To pave the way towards accurate simulation of shock/turbulent boundary-layer interaction, a series of wave and shock interaction benchmark problems that increase in complexity, are computed in this paper with triangular/tetrahedral meshes. Preliminary computations for the normal shock/turbulence interactions are carried out with a relatively coarse mesh, by direct numerical simulations standards, in order to assess other effects such as boundary conditions and the necessity of a buffer domain. The results indicate that qualitative agreement with previous studies can be obtained for flows where, strong shocks co-exist along with unsteady waves that display a broad range of scales, with a relatively compact computational domain and less stringent requirements for grid clustering near the shock. With the space-time conservation properties, stable solutions without any spurious wave reflections can be obtained without a need for buffer domains near the outflow/farfield boundaries. Computational results for the isotropic turbulent flow decay, at a relatively high turbulent Mach number, show a nicely behaved spectral decay rate for medium to high wave numbers. The high-order CESE schemes offer very robust solutions even with the presence of strong shocks or widespread shocklets. The explicit formulation in conjunction with a close to unity theoretical upper Courant number bound has the potential to offer an efficient numerical framework for general compressible turbulent flow simulations with unstructured meshes.

Chang, Chau-Lyan↗

Asymmetric electroosmotic flow and mobility measurements at nonstationary positions in the rectangular chamber

The electrophoretic mobility of a cell in solution is defined by its velocity divided by the electric field strength it experiences. An obvious way to measure the mobility of cells is to apply a constant electric field to a suspension of cells in a glass chamber and clock the velocities of individual cells through a microscope. This microscope method is the classic technique in cell electrophoresis and it has been used for the bulk of research in this field. Two aspects of the microscope method can critically affect the accuracy and consistency of its cell mobility measurements: (1) the electroosmotic fluctuations in the chamber from measurement to measurement; and (2) the number of cells which can be practically measured for statistically meaningful results. A new method of analyzing microelectrophoretic data using a computer program has been developed which addresses both of these aspects. It makes possible the mobility measurements of individual cells as positions throughout the rectangular chamber depth during asymmetric electroosmotic flow.

Source record↗

Flow-dependent regulation of endothelial nitric oxide synthase: role of protein kinases

Vascular endothelial cells are directly and continuously exposed to fluid shear stress generated by blood flow. Shear stress regulates endothelial structure and function by controlling expression of mechanosensitive genes and production of vasoactive factors such as nitric oxide (NO). Though it is well known that shear stress stimulates NO production from endothelial nitric oxide synthase (eNOS), the underlying molecular mechanisms remain unclear and controversial. Shear-induced production of NO involves Ca2+/calmodulin-independent mechanisms, including phosphorylation of eNOS at several sites and its interaction with other proteins, including caveolin and heat shock protein-90. There have been conflicting results as to which protein kinases-protein kinase A, protein kinase B (Akt), other Ser/Thr protein kinases, or tyrosine kinases-are responsible for shear-dependent eNOS regulation. The functional significance of each phosphorylation site is still unclear. We have attempted to summarize the current status of understanding in shear-dependent eNOS regulation.

Non-NASA Center↗

Multigrid schemes for viscous hypersonic flows

Several multigrid schemes are considered for the numerical computation of viscous hypersonic flows. For each scheme, the basic solution algorithm employs upwind spatial discretization with explicit multistage time stepping. Two-level versions of the various multigrid algorithms are applied to the two-dimensional advection equation, and Fourier analysis is used to determine their damping properties. The capabilities of the multigrid methods are assessed by solving two different hypersonic flow problems. Some new multigrid schemes, based on semicoarsening strategies, are shown to be quite effective in relieving the stiffness caused by the high-aspect-ratio cells required to resolve high Reynolds number flows. These schemes exhibit good convergence rates for Reynolds numbers up to 200 x 10(exp 6).

Swanson, R. C.↗

Method and Apparatus for a Miniature Bioreactor System for Long-Term Cell Culture

A bioreactor and method that permits continuous and simultaneous short, moderate, or long term cell culturing of one or more cell types or tissue in a laminar flow configuration is disclosed, where the bioreactor supports at least two laminar flow zones, which are isolated by laminar flow without the need for physical barriers between the zones. The bioreactors of this invention are ideally suited for studying short, moderate and long term studies of cell cultures and the response of cell cultures to one or more stressors such as pharmaceuticals, hypoxia, pathogens, or any other stressor. The bioreactors of this invention are also ideally suited for short, moderate or long term cell culturing with periodic cell harvesting and/or medium processing for secreted cellular components.

Kleis, Stanley J.↗

Altered cytokine production by specific human peripheral blood cell subsets immediately following space flight

In this study, flow cytometry was used to positively identify the specific lymphocyte subsets exhibiting space flight-induced alterations in cytokine production. Whole blood samples were collected from 27 astronauts at three points (one preflight, two postflight) surrounding four space shuttle missions. Assays performed included serum/urine stress hormones, white blood cell (WBC) phenotyping, and intracellular cytokine production following mitogenic stimulation. Absolute levels of peripheral granulocytes were significantly elevated following space flight, but the levels of circulating lymphocytes and monocytes were unchanged. Lymphocyte subset analysis demonstrated a decreased percentage of T cells, whereas percentages of B cells and natural killer (NK) cells remained unchanged after flight. Nearly all the astronauts exhibited an increased CD4/CD8 T cell ratio. Assessment of naive (CD45RA+) vs. memory (CD45RO+) CD4+ T cell subsets was ambiguous, and subjects tended to group within specific missions. Although no significant trend was seen in absolute monocyte levels, a significant decrease in the percentage of the CD14+ CD16+ monocytes was seen following space flight in all subjects tested. T cell (CD3+) production of interleukin-2 (IL-2) was significantly decreased after space flight, as was IL-2 production by both CD4+ and CD8+ T cell subsets. Production of interferon-gamma (IFN-gamma) was not altered by space flight for the CD8+ cell subset, but there was a significant decrease in IFN-gamma production for the CD4+ T cell subset. Serum and urine stress hormone analysis indicated significant physiologic stresses in astronauts following space flight. Altered peripheral leukocyte subsets, altered serum and urine stress hormone levels, and altered T cell cytokine secretion profiles were all observed postflight. In addition, there appeared to be differential susceptibility to space flight regarding cytokine secretion by T cell subsets. These alterations may be the result of either microgravity exposure or the physiologic stresses of landing and readaptation to unit gravity. Future studies, including in-flight analysis or sampling, will be necessary to determine the cause of these alterations.

NASA Discipline Cell Biology↗

Oxygen plant breadboard design, and techniques for improving mission figure-of-merit

A breadboard oxygen plant to process anaerobic carbon dioxide is designed and constructed; the objective is not only to produce a key propellant component extraterrestrially, but also to develop the important technologies that are necessary for a successful operation of in-situ materials utilization hardware. The solid electrolytic cells are supplied to specifications by an established vendor. The cell thermal control, electrical control, and flow control are installed after detailed designs. Extensive data are obtained that characterize the operation of the plant as the input parameters are varied. The initial mass, energy, and volume-needs provide the input to a figure-of-merit software program to calculate the impact of various candidate technologies upon the overall mission. The desirability of studies on storage and high-density propellants is shown. This task dovetails into other tasks that are evaluating alternative cell materials, catalysis for compactness, and smart sensors for effective control.

Ramohalli, Kumar↗

The interaction of a magnetic cloud with the Earth - Ionospheric convection in the Northern and Southern Hemispheres for a wide range of quasi-steady interplanetary magnetic field conditions

Observations are presented of the ionospheric convection in cross sections of the polar cap and auroral zone as part of the study of the interaction of the Earth's magnetosphere with the magnetic cloud of January 13-15, 1988. For strongly northward IMF, the convection in the Southern Hemisphere is characterized by a two-cell convection pattern comfined to high latitudes with sunward flow over the pole. The strength of the flows is comparable to that later seen under southward IMF. Superimposed on this convection pattern there are clear dawn-dusk asymmetries associated with a one-cell convection component whose sense depends on the polarity of the magnetic cloud's large east-west magnetic field component. When the cloud's magnetic field turns southward, the convection is characterized by a two-cell pattern extending to lower latitude with antisunward flow over the pole. There is no evident interhemispheric difference in the structure and strength of the convection. Superimposed dawn-dusk asymmetries in the flow patterns are observed which are only in part attributable to the east-west component of the magnetic field.

Freeman, M. P.↗

Numerical Models of Human Circulatory System under Altered Gravity: Brain Circulation

A computational fluid dynamics (CFD) approach is presented to model the blood flow through the human circulatory system under altered gravity conditions. Models required for CFD simulation relevant to major hemodynamic issues are introduced such as non-Newtonian flow models governed by red blood cells, a model for arterial wall motion due to fluid-wall interactions, a vascular bed model for outflow boundary conditions, and a model for auto-regulation mechanism. The three-dimensional unsteady incompressible Navier-Stokes equations coupled with these models are solved iteratively using the pseudocompressibility method and dual time stepping. Moving wall boundary conditions from the first-order fluid-wall interaction model are used to study the influence of arterial wall distensibility on flow patterns and wall shear stresses during the heart pulse. A vascular bed modeling utilizing the analogy with electric circuits is coupled with an auto-regulation algorithm for multiple outflow boundaries. For the treatment of complex geometry, a chimera overset grid technique is adopted to obtain connectivity between arterial branches. For code validation, computed results are compared with experimental data for steady and unsteady non-Newtonian flows. Good agreement is obtained for both cases. In sin-type Gravity Benchmark Problems, gravity source terms are added to the Navier-Stokes equations to study the effect of gravitational variation on the human circulatory system. This computational approach is then applied to localized blood flows through a realistic carotid bifurcation and two Circle of Willis models, one using an idealized geometry and the other model using an anatomical data set. A three- dimensional anatomical Circle of Willis configuration is reconstructed from human-specific magnetic resonance images using an image segmentation method. The blood flow through these Circle of Willis models is simulated to provide means for studying gravitational effects on the brain circulation under auto-regulation.

Kim, Chang Sung↗