Setting of graded levels of a protein in yeast by a t-degron technique as applied to phosphoglycerate mutase

BACKGROUND: Setting of graded levels of a protein for in vivo studies by controlled gene expression has inconveniences, and we here explore the use of the t-degron technique instead. RESULTS: In a yeast t-degron (ubiquitin-argDHFR(ts))- phosphoglycerate mutase (GPM1) fusion strain, increasing periods of exposure to the non-permissive temperature 37°C, even in the presence of cycloheximide, gave decreasing function, as assessed at 23°C in vivo by glucose metabolism and confirmed by immunoblot. CONCLUSION: An ideal system would set a range of lower levels of a protein, do so without compensating protein synthesis, and give stable activity for in vitro comparisons. Although the first two aims appear obtainable, the third was not in this example of the application, limiting its uses for some but not all purposes

Words with the Maximum Number of Abelian Squares

An abelian square is the concatenation of two words that are anagrams of one another. A word of length $n$ can contain $\Theta(n^2)$ distinct factors that are abelian squares. We study infinite words such that the number of abelian square factors of length $n$ grows quadratically with $n$.Comment: To appear in the proceedings of WORDS 201

Advanced experimental applications for x-ray transmission gratings Spectroscopy using a novel grating fabrication method

A novel fabrication method for soft x-ray transmission grating and other optical elements is presented. The method uses Focused-Ion-Beam (FIB) technology to fabricate high-quality free standing grating bars on Transmission Electron Microscopy grids (TEM-grid). High quality transmission gratings are obtained with superb accuracy and versatility. Using these gratings and back-illuminated CCD camera, absolutely calibrated x-ray spectra can be acquired for soft x-ray source diagnostics in the 100-3000 eV spectral range. Double grating combinations of identical or different parameters are easily fabricated, allowing advanced one-shot application of transmission grating spectroscopy. These applications include spectroscopy with different spectral resolutions, bandwidths, dynamic ranges, and may serve for identification of high-order contribution, and spectral calibrations of various x-ray optical elements

A chemical screen identifies small molecules that regulate hepcidin expression

AbstractHepcidin, a peptide hormone produced in the liver, decreases intestinal iron absorption and macrophage iron release via effects on ferroportin. Bone morphogenic protein and Stat3 signaling regulate Hepcidin's transcription. Hepcidin is a potential drug target for patients with iron overload syndromes because its levels are inappropriately low in these individuals. To generate a tool for identifying small molecules that modulate Hepcidin expression, we stably transfected human hepatocytes (HepG2) cells with a reporter construct containing 2.7kb of the human Hepcidin promoter upstream of a firefly reporter gene. We used high throughput methods to screen 10,169 chemicals in duplicate for their effect on Hepcidin expression and cell viability. Regulators were identified as chemicals that caused a change >3 standard deviations above or >1 standard deviation below the mean of the other chemicals (z-score >3 or <1), while not adversely affecting cell viability, quantified by fluorescence assay. Following validation assays, we identified 16 chemicals in a broad range of functional classes that promote Hepcidin expression. All of the chemicals identified increased expression of bone morphogenic protein-dependent and/or Stat3-dependent genes, however none of them strongly increased phosphorylation of Smad1,5,8 or Stat3

A pedagogical example: a family of stochastic cellular automata that plays Alesia

International audienceAlesia is a two-player zero-sum game which is quite similar to the rock-paper-scissors game: the two players simultaneously move and do not know what the opponent plays at given round. The simultaneity of the moves implies that there is no deterministic good strategy in this game, otherwise one would anticipate the moves of the opponent and easily win the game. We explore how to build a family of one-dimensional stochastic cellular automata to play this game. The rules are built in an iterative way by progressively increasing the complexity of the transitions. We show the possibility to construct a family of rules with interesting results, including a good performance when confronted to the Nash-equilibrium strategy

Predicting the deleterious effects of mutation load in fragmented populations.

Human-induced habitat fragmentation constitutes a major threat to biodiversity. Both genetic and demographic factors combine to drive small and isolated populations into extinction vortices. Nevertheless, the deleterious effects of inbreeding and drift load may depend on population structure, migration patterns, and mating systems and are difficult to predict in the absence of crossing experiments. We performed stochastic individual-based simulations aimed at predicting the effects of deleterious mutations on population fitness (offspring viability and median time to extinction) under a variety of settings (landscape configurations, migration models, and mating systems) on the basis of easy-to-collect demographic and genetic information. Pooling all simulations, a large part (70%) of variance in offspring viability was explained by a combination of genetic structure (F(ST)) and within-deme heterozygosity (H(S)). A similar part of variance in median time to extinction was explained by a combination of local population size (N) and heterozygosity (H(S)). In both cases the predictive power increased above 80% when information on mating systems was available. These results provide robust predictive models to evaluate the viability prospects of fragmented populations