Analysis of metabolic flux distributions in relation to the extracellular environment in Avian cells

Continuous cell lines that proliferate in chemically defined and simple media have been highly regarded as suitable alternatives for vaccine production. One such cell line is the AG1.CR.pIX avian cell line developed by PROBIOGEN. This cell line can be cultivated in a fully scalable suspension culture and adapted to grow in chemically defined, calf serum free, medium [1]–[5]. The medium composition and cultivation strategy are important factors for reaching high virus titers. In this project, a series of computational methods was used to simulate the cell’s response to different environments. The study is based on the metabolic model of the central metabolism proposed in [1]. In a first step, Metabolic Flux Analysis (MFA) was used along with measured uptake and secretion fluxes to estimate intracellular flux values. The network and data were found to be consistent. In a second step, Flux Balance Analysis (FBA) was performed to access the cell’s biological objective. The objective that resulted in the best predicted results fit to the experimental data was the minimization of oxidative phosphorylation. Employing this objective, in the next step Flux Variability Analysis (FVA) was used to characterize the flux solution space. Furthermore, various scenarios, where a reaction deletion (elimination of the compound from the media) was simulated, were performed and the flux solution space for each scenario was calculated. Growth restrictions caused by essential and non-essential amino acids were accurately predicted. Fluxes related to the essential amino acids uptake and catabolism, the lipid synthesis and ATP production via TCA were found to be essential to exponential growth. Finally, the data gathered during the previous steps were analyzed using principal component analysis (PCA), in order to assess potential changes in the physiological state of the cell. Three metabolic states were found, which correspond to zero, partial and maximum biomass growth rate. Elimination of non-essential amino acids or pyruvate from the media showed no impact on the cell’s assumed normal metabolic state.

Surface-dependent strategies and energy flux in benthic marine communities or, why corals do not exist in the Mediterranean

Most structure-building organisms in rocky benthic communities are surface-dependent because their energy inputs depend mainly on the surface they expose to water. Two photosynthetic strategies, divided into calcareous and non calcareous algae, strict suspension-feeders and photosynthetic suspension feeders (e.g. hermatypic corals) are the four main strategies evolutively acquired by benthic organisms. Competition between those strategies occur in relation to productivity of the different species, in such a way that, for given environmental conditions, species with a higher growth (P/B ratio) would dominate. At a worldwide scale, littoral marine benthos can he considered to fit into the four fields defined by two main axes: the first, relates to productivity and relies atrophic and oligotrophic waters and the second is defined by the degree of environmental variability or seasonality (from high to low). Coral reefs (marine ecosystems dominated by photosynthetic suspension feeders) develop in the space of oligotrophic areas with low variability, while kelp beds (marine ecosystem dominated by large, non calcareous algae) are to be found only in eutrophic places with a high variability. The space of eutrophic waters with a low variability do not has specially adapted, high structured, benthic marine ecosystems, and in these conditions opportunistic algae and animals predominate. Finally, photophilic mediterranean benthos -devoid of kelps and without hermatypic corals- typifies the field of oligotrophic areas with high variability; in its more genuine aspect, Mediterranean benthos is represented by small algae with a high percentage of calcareous thallii. In all cases strict suspension-feeders compete successfully with photosynthetic organisms only in situations of low irradiances or very high inputs of POM. In its turn, Mediterranean rocky benthos, in spite of its relative uniformity, is geographically organized along the same axes. The Gulf of Lions and the insular bottoms (Balearic Islands, for example) would correspond to the extremes of eutrophic-high variability areas and oligotrophic-low variability areas, respectively. Irradiance, nutrient and POM concentration, and hydrodynamism are the three variables which mainly affect the distribution of the different surface-dependent strategies, and thus, these parameters are of paramount interest for understanding the trophic structure of Mediterranean benthic communities. In environments non limited by light, nutrient availability, defined as the product between nutrient -POM concentration and hydrodynamism, states the dominance of calcareous versus non calcareous algae. Calcareous algae dominate in oligotrophic waters while non-calcareous algae dominate in moderately eutrophic waters. In light-limited environments, passive suspension feeders (octocorallaria, gorgonians) become dominant species if POM availability is enhanced by a high hydrodynamism (strong currents); in waters with a low charge of POM organisms of other groups, mainly active suspension feeders, predominate (sponges, bryozoans, scleractiniarians). In any case, there always exists a very variable bathymetric zone, depending on light attenuation and nutrient-POM availability, where encrusting calcareous algae strongly compete with suspension feeders (coralligenous).

Do magnetospheric shear Alfvén waves generate sufficient electron energy flux to power the aurora?

Using a self-consistent drift-kinetic simulation code, we investigate whether electron acceleration owing to shear Alfvén waves in the plasma sheet boundary layer is sufficient to cause auroral brightening in the ionosphere. The free parameters used in the simulation code are guided by in situ observations of wave and plasma parameters in the magnetosphere at distances >4 RE from the Earth. For the perpendicular wavelength used in the study, which maps to ∼4 km at 110 km altitude, there is a clear amplitude threshold which determines whether magnetospheric shear Alfvén waves above the classical auroral acceleration region can excite sufficient electrons to create the aurora. Previous studies reported wave amplitudes that easily exceed this threshold; hence, the results reported in this paper demonstrate that auroral acceleration owing to shear Alfvén waves can occur in the magnetosphere at distances >4 RE from the Earth.

Characterization of energy flux partitioning in urban environments: links with surface seasonal properties

A better understanding of links between the properties of the urban environment and the exchange to the atmosphere is central to a wide range of applications. The numerous measurements of surface energy balance data in urban areas enable intercomparison of observed fluxes from distinct environments. This study analyzes a large database in two new ways. First, instead of normalizing fluxes using net all-wave radiation only the incoming radiative fluxes are used, to remove the surface attributes from the denominator. Second, because data are now available year-round, indices are developed to characterize the fraction of the surface (built; vegetation) actively engaged in energy exchanges. These account for shading patterns within city streets and seasonal changes in vegetation phenology; their impact on the partitioning of the incoming radiation is analyzed. Data from 19 sites in North America, Europe, Africa, and Asia (including 6-yr-long observation campaigns) are used to derive generalized surface–flux relations. The midday-period outgoing radiative fraction decreases with an increasing total active surface index, the stored energy fraction increases with an active built index, and the latent heat fraction increases with an active vegetated index. Parameterizations of these energy exchange ratios as a function of the surface indices [i.e., the Flux Ratio–Active Index Surface Exchange (FRAISE) scheme] are developed. These are used to define four urban zones that characterize energy partitioning on the basis of their active surface indices. An independent evaluation of FRAISE, using three additional sites from the Basel Urban Boundary Layer Experiment (BUBBLE), yields accurate predictions of the midday flux partitioning at each location.

Movable-bed laboratory experiments comparing radiation stress and energy flux factor as predictors of longshore transport rate /

"April 1981."

Computation of longshore energy flux using LEO current observations /

"March 1980."

The SPM energy flux method for predicting longshore transport rate /

"June 1980."

Computer algorithm to calculate longshore energy flux and wave direction from a two pressure sensor array /

"August 1982."

The temporal evolution of the energy flux across scales in homogeneous turbulence

A temporal study of energy transfer across length scales is performed in 3D numerical simulations of homogeneous shear flow and isotropic turbulence. The average time taken by perturbations in the energy flux to travel between scales is measured and shown to be additive. Our data suggests that the propagation of disturbances in the energy flux is independent of the forcing and that it defines a ‘velocity’ that determines the energy flux itself. These results support that the cascade is, on average, a scale-local process where energy is continuously transmitted from one scale to the next in order of decreasing size.

Distance Distributions and Energy of Designs in Hamming Spaces

We obtain new combinatorial upper and lower bounds for the potential energy of designs in q-ary Hamming space. Combined with results on reducing the number of all feasible distance distributions of such designs this gives reasonable good bounds. We compute and compare our lower bounds to recently obtained universal lower bounds. Some examples in the binary case are considered.

Algorithms and computational tools for metabolic flux analysis

PhD thesis in Biomedical Engineering

Spectral nonlocality, absolute equilibria and Kraichnan-Leith-Batchelor phenomenology in two-dimensional turbulent energy cascades

We study the degree to which Kraichnan–Leith–Batchelor (KLB) phenomenology describes two-dimensional energy cascades in α turbulence, governed by ∂θ/∂t+J(ψ,θ)=ν∇2θ+f, where θ=(−Δ)α/2ψ is generalized vorticity, and ψ^(k)=k−αθ^(k) in Fourier space. These models differ in spectral non-locality, and include surface quasigeostrophic flow (α=1), regular two-dimensional flow (α=2) and rotating shallow flow (α=3), which is the isotropic limit of a mantle convection model. We re-examine arguments for dual inverse energy and direct enstrophy cascades, including Fjørtoft analysis, which we extend to general α, and point out their limitations. Using an α-dependent eddy-damped quasinormal Markovian (EDQNM) closure, we seek self-similar inertial range solutions and study their characteristics. Our present focus is not on coherent structures, which the EDQNM filters out, but on any self-similar and approximately Gaussian turbulent component that may exist in the flow and be described by KLB phenomenology. For this, the EDQNM is an appropriate tool. Non-local triads contribute increasingly to the energy flux as α increases. More importantly, the energy cascade is downscale in the self-similar inertial range for 2.5<α<10. At α=2.5 and α=10, the KLB spectra correspond, respectively, to enstrophy and energy equipartition, and the triad energy transfers and flux vanish identically. Eddy turnover time and strain rate arguments suggest the inverse energy cascade should obey KLB phenomenology and be self-similar for α<4. However, downscale energy flux in the EDQNM self-similar inertial range for α>2.5 leads us to predict that any inverse cascade for α≥2.5 will not exhibit KLB phenomenology, and specifically the KLB energy spectrum. Numerical simulations confirm this: the inverse cascade energy spectrum for α≥2.5 is significantly steeper than the KLB prediction, while for α<2.5 we obtain the KLB spectrum.

Energy exchange in a dense urban environment – part I: temporal variability of long-term observations in central London

Though anthropogenic impacts on boundary layer climates are expected to be large in dense urban areas, to date very few studies of energy flux observations are available. We report on 3.5 years of measurements gathered in central London, UK. Radiometer and eddy covariance observations at two adjacent sites, at different heights, were analysed at various temporal scales and with respect to meteorological conditions, such as cloud cover. Although the evaporative flux is generally small due to low moisture availability and a predominately impervious surface, the enhancement following rainfall usually lasts for 12–18 h. As both the latent and sensible heat fluxes are larger in the afternoon, they maintain a relatively consistent Bowen ratio throughout the middle of the day. Strong storage and anthropogenic heat fluxes sustain high and persistently positive sensible heat fluxes. At the monthly time scale, the urban surface often loses more energy by this turbulent heat flux than is gained from net all-wave radiation. Auxiliary anthropogenic heat flux information suggest human activities in the study area are sufficient to provide this energy.

A comparison between ion characteristics observed by the POLAR and DMSP spacecraft in the high-latitude magnetosphere

We study here the injection and transport of ions in the convection-dominated region of the Earth’s magnetosphere. The total ion counts from the CAMMICE MICS instrument aboard the POLAR spacecraft are used to generate occurrence probability distributions of magnetospheric ion populations. MICS ion spectra are characterised by both the peak in the differential energy flux, and the average energy of ions striking the detector. The former permits a comparison with the Stubbs et al. (2001) survey of He2+ ions of solar wind origin within the magnetosphere. The latter can address the occurrences of various classifications of precipitating particle fluxes observed in the topside ionosphere by DMSP satellites (Newell and Meng, 1992). The peak energy occurrences are consistent with our earlier work, including the dawn-dusk asymmetry with enhanced occurrences on the dawn flank at low energies, switching to the dusk flank at higher energies. The differences in the ion energies observed in these two studies can be explained by drift orbit effects and acceleration processes at the magnetopause, and in the tail current sheet. Near noon at average ion energies of _1 keV, the cusp and open LLBL occur further poleward here than in the Newell and Meng survey, probably due to convectionrelated time-of-flight effects. An important new result is that the pre-noon bias previously observed in the LLBL is most likely due to the component of this population on closed field lines, formed largely by low energy ions drifting earthward from the tail. There is no evidence here of mass and momentum transfer from the solar wind to the LLBL by nonreconnection coupling. At higher energies (_2–20 keV), we observe ions mapping to the auroral oval and can distinguish between the boundary and central plasma sheets. We show that ions at these energies relate to a transition from dawnward to duskward dominated flow, this is evidence of how ion drift orbits in the tail influence the location and behaviour of the plasma populations in the magnetosphere.

Combining satellite observations and reanalysis energy transports to estimate global net surface energy fluxes 1985-2012

Two methods are developed to estimate net surface energy fluxes based upon satellite-based reconstructions of radiative fluxes at the top of atmosphere and the atmospheric energy tendencies and transports from the ERA-Interim reanalysis. Method 1 applies the mass adjusted energy divergence from ERA-Interim while method 2 estimates energy divergence based upon the net energy difference at the top of atmosphere and the surface from ERA-Interim. To optimise the surface flux and its variability over ocean, the divergences over land are constrained to match the monthly area mean surface net energy flux variability derived from a simple relationship between the surface net energy flux and the surface temperature change. The energy divergences over the oceans are then adjusted to remove an unphysical residual global mean atmospheric energy divergence. The estimated net surface energy fluxes are compared with other data sets from reanalysis and atmospheric model simulations. The spatial correlation coefficients of multi-annual means between the estimations made here and other data sets are all around 0.9. There are good agreements in area mean anomaly variability over the global ocean, but discrepancies in the trend over the eastern Pacific are apparent.