Efficient cloning system for construction of gene silencing vectors in Aspergillus niger

An approach based on Gateway recombination technology to efficiently construct silencing vectors was developed for use in the biotechnologically important fungus Aspergillus niger. The transcription activator of xylanolytic and cellulolytic genes XlnR of A. niger was chosen as target for gene silencing. Silencing was based on the expression vector pXLNRir that was constructed and used in co-transformation. From all the strains isolated (N = 77), nine showed poor xylan-degrading activities in two semi-quantitative plate assays testing different activities for xylan degradation. Upon induction on D-xylose, transcript levels of xlnR were decreased in the xlnR-silenced strains, compared to a wild-type background. Under these conditions, the transcript levels of xyrA and xynB (two genes regulated by XlnR) were also decreased for these xlnR-silenced strains. These results indicate that the newly developed system for rapid generation of silencing vectors is an effective tool for A. niger, and this can be used to generate strains with a tailored spectrum of enzyme activities or product formation by silencing specific genes encoding, e.g., regulators such as Xln

Shotgun proteomics of Aspergillus niger microsomes upon D-xylose induction

Protein secretion plays an eminent role in cell maintenance and adaptation to the extracellular environment of microorganisms. Although protein secretion is an extremely efficient process in filamentous fungi, the mechanisms underlying protein secretion have remained largely uncharacterized in these organisms. In this study, we analyzed the effects of the d-xylose induction of cellulase and hemicellulase enzyme secretion on the protein composition of secretory organelles in Aspergillus niger. We aimed to systematically identify the components involved in the secretion of these enzymes via mass spectrometry of enriched subcellular microsomal fractions. Under each condition, fractions enriched for secretory organelles were processed for tandem mass spectrometry, resulting in the identification of peptides that originate from 1,081 proteins, 254 of which-many of them hypothetical proteins-were predicted to play direct roles in the secretory pathway. d-Xylose induction led to an increase in specific small GTPases known to be associated with polarized growth, exocytosis, and endocytosis. Moreover, the endoplasmic-reticulum-associated degradation (ERAD) components Cdc48 and all 14 of the 20S proteasomal subunits were recruited to the secretory organelles. In conclusion, induction of extracellular enzymes results in specific changes in the secretory subproteome of A. niger, and the most prominent change found in this study was the recruitment of the 20S proteasomal subunits to the secretory organelle

Morris-Thorne wormholes with a cosmological constant

First, the ideas introduced in the wormhole research field since the work of Morris and Thorne are briefly reviewed, namely, the issues of energy conditions, wormhole construction, stability, time machines and astrophysical signatures. Then, spherically symmetric and static traversable Morris-Thorne wormholes in the presence of a generic cosmological constant are analyzed. A matching of an interior solution to the unique exterior vacuum solution is done using directly the Einstein equations. The structure as well as several physical properties and characteristics of traversable wormholes due to the effects of the cosmological term are studied. Interesting equations appear in the process of matching. For instance, one finds that for asymptotically flat and anti-de Sitter spacetimes the surface tangential pressure of the thin-shell, at the boundary of the interior and exterior solutions, is always strictly positive, whereas for de Sitter spacetime it can take either sign as one could expect, being negative (tension) for relatively high cosmological constant and high wormhole radius, positive for relatively high mass and small wormhole radius, and zero in-between. Finally, some specific solutions with generic cosmological constant, based on the Morris-Thorne solutions, are provided.Comment: latex, 49 pages, 8 figures. Expanded version of the paper published in Physical Review

New Upper Limit of Terrestrial Equivalence Principle Test for Rotating Extended Bodies

Improved terrestrial experiment to test the equivalence principle for rotating extended bodies is presented, and a new upper limit for the violation of the equivalence principle is obtained at the level of 1.6$10^{\text{-7}}$, which is limited by the friction of the rotating gyroscope. It means the spin-gravity interaction between the extended bodies has not been observed at this level.Comment: 4 page

Thermal Density Functional Theory in Context

This chapter introduces thermal density functional theory, starting from the ground-state theory and assuming a background in quantum mechanics and statistical mechanics. We review the foundations of density functional theory (DFT) by illustrating some of its key reformulations. The basics of DFT for thermal ensembles are explained in this context, as are tools useful for analysis and development of approximations. We close by discussing some key ideas relating thermal DFT and the ground state. This review emphasizes thermal DFT's strengths as a consistent and general framework.Comment: Submitted to Spring Verlag as chapter in "Computational Challenges in Warm Dense Matter", F. Graziani et al. ed

Gravitational field around a time-like current-carrying screwed cosmic string in scalar-tensor theories

In this paper we obtain the space-time generated by a time-like current-carrying superconducting screwed cosmic string(TCSCS). This gravitational field is obtained in a modified scalar-tensor theory in the sense that torsion is taken into account. We show that this solution is comptible with a torsion field generated by the scalar field $\phi$. The analysis of gravitational effects of a TCSCS shows up that the torsion effects that appear in the physical frame of Jordan-Fierz can be described in a geometric form given by contorsion term plus a symmetric part which contains the scalar gradient. As an important application of this solution, we consider the linear perturbation method developed by Zel'dovich, investigate the accretion of cold dark matter due to the formation of wakes when a TCSCS moves with speed $v$ and discuss the role played by torsion. Our results are compared with those obtained for cosmic strings in the framework of scalar-tensor theories without taking torsion into account.Comment: 21 pages, no figures, Revised Version, presented at the "XXIV- Encontro Nacional de Fisica de Particulas e Campos ", Caxambu, MG, Brazil, to appear in Phys. Rev.