Temperature limited fed-batch technique for control of proteolysis in Pichia pastoris bioreactor cultures

BACKGROUND: A temperature limited fed-batch (TLFB) technique is described and used for Pichia pastoris Mut(+ )strain cultures and compared with the traditional methanol limited fed-batch (MLFB) technique. A recombinant fusion protein composed of a cellulose-binding module (CBM) from Neocallimastix patriciarum cellulase 6A and lipase B from Candida antarctica (CALB), was produced and secreted by this strain. RESULTS: A protein concentration of about 1 g L(-1 )was produced in the MLFB process. However, this product was considerably degraded by protease(s). By applying the TLFB process, the yield was increased to 2 g L(-1 )full-length product and no proteolytic degradation was observed. Flow cytometry analysis showed that the percentage of dead cells increased rapidly during the initial methanol feed phase in the MLFB process and reached a maximum of about 12% after about 40–70 hours of methanol feeding. In the TLFB process, cell death rate was low and constant and reached 4% dead cells at the end of cultivation (about 150 hours methanol feeding time). The lower cell death rate in the TLFB correlated with a lower protease activity in the culture supernatant. The specific alcohol oxidase (AOX) activity in the TLFB process was 3.5 times higher than in the MLFB process. CONCLUSION: Three mechanisms that may contribute to the much higher accumulation of product in the TLFB process are: 1) reduced proteolysis due to lower temperature, 2) reduced proteolysis due to lower cell death and protease release to the medium, 3) increased synthesis rate due to higher AOX activity

From Heisenberg matrix mechanics to EBK quantization: theory and first applications

Despite the seminal connection between classical multiply-periodic motion and Heisenberg matrix mechanics and the massive amount of work done on the associated problem of semiclassical (EBK) quantization of bound states, we show that there are, nevertheless, a number of previously unexploited aspects of this relationship that bear on the quantum-classical correspondence. In particular, we emphasize a quantum variational principle that implies the classical variational principle for invariant tori. We also expose the more indirect connection between commutation relations and quantization of action variables. With the help of several standard models with one or two degrees of freedom, we then illustrate how the methods of Heisenberg matrix mechanics described in this paper may be used to obtain quantum solutions with a modest increase in effort compared to semiclassical calculations. We also describe and apply a method for obtaining leading quantum corrections to EBK results. Finally, we suggest several new or modified applications of EBK quantization.Comment: 37 pages including 3 poscript figures, submitted to Phys. Rev.

Hybrid Natural Low Scale Inflation

We discuss the phenomenological implications of hybrid natural inflation models in which the inflaton is a pseudo-Goldstone boson but inflation is terminated by a second scalar field. A feature of the scheme is that the scale of breaking of the Goldstone symmetry can be lower than the Planck scale and so gravitational corrections are under control. We show that, for supersymmetric models, the scale of inflation can be chosen anywhere between the Lyth upper bound and a value close to the electroweak breaking scale. Unlike previous models of low scale inflation the observed density perturbations and spectral index are readily obtained by the choice of the free parameters

A large stellar evolution database for population synthesis studies: VI. White dwarf cooling sequences

We present a new set of cooling models and isochrones for both H- and He-atmosphere white dwarfs, incorporating accurate boundary conditions from detailed model atmosphere calculations, and carbon-oxygen chemical abundance profiles based on updated stellar evolution calculations from the BaSTI stellar evolution archive - a theoretical data center for the Virtual Observatory. We discuss and quantify the uncertainties in the cooling times predicted by the models, arising from the treatment of mixing during the central H- and He-burning phases, number of thermal pulses experienced by the progenitors, progenitor metallicity and the $^{12}C(\alpha,\gamma)^{16}O$ reaction rate. The largest sources of uncertainty turn out to be related to the treatment of convection during the last stages of the progenitor central He-burning phase, and the $^{12}C(\alpha,\gamma)^{16}O$ reaction rate. We compare our new models to previous calculations performed with the same stellar evolution code, and discuss their application to the estimate of the age of the solar neighborhood, and the interpretation of the observed number ratios between H- and He-atmosphere white dwarfs. The new white dwarf sequences and an extensive set of white dwarf isochrones that cover a large range of ages and progenitor metallicities are made publicly available at the official BaSTI website.Comment: 34 pages, 13 figures, The Astrophysical Journal, in pres

Optimized clustering estimators for BAO measurements accounting for significant redshift uncertainty

We determine an optimized clustering statistic to be used for galaxy samples with significant redshift uncertainty, such as those that rely on photometric redshifts. To do so, we study the baryon acoustic oscillation (BAO) information content as a function of the orientation of galaxy clustering modes with respect to their angle to the line of sight (LOS). The clustering along the LOS, as observed in a redshift-space with significant redshift uncertainty, has contributions from clustering modes with a range of orientations with respect to the true LOS. For redshift uncertainty σz ≥ 0.02(1 + z), we find that while the BAO information is confined to transverse clustering modes in the true space, it is spread nearly evenly in the observed space. Thus, measuring clustering in terms of the projected separation (regardless of the LOS) is an efficient and nearly lossless compression of the signal for σz ≥ 0.02(1 + z). For reduced redshift uncertainty, a more careful consideration is required. We then use more than 1700 realizations (combining two separate sets) of galaxy simulations mimicking the Dark Energy Survey Year 1 (DES Y1) sample to validate our analytic results and optimized analysis procedure. We find that using the correlation function binned in projected separation, we can achieve uncertainties that are within 10 per cent of those predicted by Fisher matrix forecasts. We predict that DES Y1 should achieve a 5 per cent distance measurement using our optimized methods. We expect the results presented here to be important for any future BAO measurements made using photometric redshift data.Please visit publisher's website for further information

Clarifying the effects of interacting dark energy on linear and nonlinear structure formation processes

We present a detailed numerical study of the impact that cosmological models featuring a direct interaction between the Dark Energy component that drives the accelerated expansion of the Universe and Cold Dark Matter can have on the linear and nonlinear stages of structure formation. By means of a series of collisionless N-body simulations we study the influence that each of the different effects characterizing these cosmological models - which include among others a fifth force, a time variation of particle masses, and a velocity-dependent acceleration - separately have on the growth of density perturbations and on a series of observable quantities related to linear and nonlinear cosmic structures, as the matter power spectrum, the gravitational bias between baryons and Cold Dark Matter, the halo mass function and the halo density profiles. We perform our analysis applying and comparing different numerical approaches previously adopted in the literature, and we address the partial discrepancies recently claimed in a similar study by Li & Barrow (2010b) with respect to the first outcomes of Baldi et al. (2010), which are found to be related to the specific numerical approach adopted in the former work. Our results fully confirm the conclusions of Baldi et al. (2010) and show that when linear and nonlinear effects of the interaction between Dark Energy and Cold Dark Matter are properly disentangled, the velocity-dependent acceleration is the leading effect acting at nonlinear scales, and in particular is the most important mechanism in lowering the concentration of Cold Dark Matter halos.Comment: 14 pages, 1 Table, 6 Figures. MNRAS accepte

CFHTLS weak-lensing constraints on the neutrino masses

We use measurements of cosmic shear from CFHTLS, combined with WMAP-5 cosmic microwave background anisotropy data, baryonic acoustic oscillations from SDSS and 2dFGRS and supernovae data from SNLS and Gold-set, to constrain the neutrino mass. We obtain a 95% confidence level upper limit of 0.54 eV for the sum of the neutrino masses, and a lower limit of 0.03 eV. The preference for massive neutrinos vanishes when shear-measurement systematics are included in the analysis.Comment: 10 pages. Published versio

The Impact of an Extra Background of Relativistic Particles on the Cosmological Parameters derived from Microwave Background Anisotropies

Recent estimates of cosmological parameters derived from Cosmic Microwave Background (CMB) anisotropies are based on the assumption that we know the precise amount of energy density in relativistic particles in the universe, $\omega_{rel}$, at all times. There are, however, many possible mechanisms that can undermine this assumption. In this paper we investigate the effect that removing this assumption has on the determination of the various cosmological parameters. We obtain fairly general bounds on the redshift of equality, $z_{eq}= \omega_{m}/\omega_{rel}=3100_{-400}^{+600}$. We show that $\omega_{rel}$ is nearly degenerate with the amount of energy in matter, $\omega_m$, and that its inclusion in CMB parameter estimation also affects the present constraints on other parameters such as the curvature or the scalar spectral index of primordial fluctuations. This degeneracy has the effect of limiting the precision of parameter estimation from the MAP satellite, but it can be broken by measurements on smaller scales such as those provided by the Planck satellite mission.Comment: 9 pages, 6 figures, Submitted to MNRAS. High resolution colour pictures can be obtained on request from the author

Tests of model predictions for the responses of stellar spectra and absorption-line indices to element abundance variations. Tests of model predictions for the responses of stellar spectra and absorption-line indices to element abundance variations.

A method that is widely used to analyse stellar populations in galaxies is to apply the theoretically derived responses of stellar spectra and line indices to element abundance variations, which are hereafter referred to as response functions. These are applied in a differential way, to base models, in order to generate spectra or indices with different abundance patterns. In this paper, sets of such response functions for three different stellar evolutionary stages are tested with new empirical [Mg/Fe] abundance data for the medium-resolution Isaac Newton Telescope library of empirical spectra (MILES). Recent theoretical models and observations are used to investigate the effects of [Fe/H], [Mg/H] and overall [Z/H] on spectra, via ratios of spectra for similar stars. The global effects of changes in abundance patterns are investigated empirically through direct comparisons of similar stars from MILES, highlighting the impact of abundance effects in the blue part of the spectrum, particularly for lower temperature stars. It is found that the relative behaviour of iron-sensitive line indices are generally well predicted by response functions, whereas Balmer line indices are not. Other indices tend to show large scatter about the predicted mean relations. Implications for element abundance and age studies in stellar populations are discussed and ways forward are suggested to improve the match with the behaviour of spectra and line-strength indices observed in real stars