Light Dominates Peripheral Circadian Oscillations in Drosophila melanogaster During Sensory Conflict

In Drosophila, as in other animals, the circadian clock is a singular entity in name and concept only. In reality, clock functions emerge from multiple processes and anatomical substrates. One distinction has conventionally been made between a central clock (in the brain) and peripheral clocks (e.g., in the gut and the eyes). Both types of clock generate robust circadian oscillations, which do not require external input. Furthermore, the phases of these oscillations remain exquisitely sensitive to specific environmental cues, such as the daily changes of light and temperature. When these cues conflict with one another, the central clock displays complex forms of sensory integration; how peripheral clocks respond to conflicting input is unclear. We therefore explored the effects of light and temperature misalignments on peripheral clocks. We show that under conflict, peripheral clocks preferentially synchronize to the light stimulus. This photic dominance requires the presence of the circadian photoreceptor, Cryptochrome

Proteomic identification of s-nitrosylated proteins in arabidopsis thaliana in response to pathogen infection

Comunicaciones a congreso

Increasing the sensivity of proteomic identification of s-nitrosylated proteins: towards the S-nitrosoproteomes in (patho)physiological settings

Comunicaciones a congreso

Development of an All-in-One Lentiviral Vector System Based on the Original TetR for the Easy Generation of Tet-ON Cell Lines

Lentiviral vectors (LVs) are considered one of the most promising vehicles to efficiently deliver genetic information for basic research and gene therapy approaches. Combining LVs with drug-inducible expression systems should allow tight control of transgene expression with minimal side effect on relevant target cells. A new doxycycline-regulated system based on the original TetR repressor was developed in 1998 as an alternative to the TetR-VP16 chimeras (tTA and rtTA) to avoid secondary effects due to the expression of transactivator domains. However, previously described TetR-based systems required cell cloning and/or antibiotic selection of tetracycline-responsive cells in order to achieve good regulation. In the present manuscript we have constructed a dual Tet-ON system based on two lentiviral vectors, one expressing the TetR through the spleen focus forming virus (SFFV) promoter (STetR) and a second expressing eGFP through the regulatable CMV-TetO promoter (CTetOE). Using these vectors we have demonstrated that the TetR repressor, contrary to the reverse transactivator (rtTA), can be expressed in excess to bind and modulate a high number of TetO operons. We have also showed that this dual vector system can generate regulatable bulk cell lines (expressing high levels of TetR) that are able to modulate transgene expression either by varying doxycycline concentration and/or by varying the amount of CTetOE vector genomes per cell. Based on these results we have developed a new all-in-one lentiviral vector (CEST) driving the expression of TetR through the SFFV promoter and the expression of eGFP through the doxycycline-responsive CMV-TetO operon. This vector efficiently produced Tet-ON regulatable immortalized (293T) and primary (human mesenchymal stem cells and human primary fibroblasts) cells. Bulk doxycycline-responsive cell lines express high levels of the transgene with low amount of doxycycline and are phenotypically indistinct from its parental cells

Quantifying variation in the ability of yeasts to attract Drosophila melanogaster

Yeasts that invade and colonise fruit significantly enhance the volatile chemical diversity of this ecosystem. These modified bouquets are thought to be more attractive to Drosophila flies than the fruit alone, but the variance of attraction in natural yeast populations is uncharacterised. Here we investigate how a range of yeast isolates affect the attraction of female D. melanogaster to fruit in a simple two choice assay comparing yeast to sterile fruit. Of the 43 yeast isolates examined, 33 were attractive and seven repellent to the flies. The results of isolate-versus-isolate comparisons provided the same relative rankings. Attractiveness varied significantly by yeast, with the strongly fermenting Saccharomyces species generally being more attractive than the mostly respiring non-Saccharomyces species (P = 0.0035). Overall the habitat (fruit or other) from which the isolates were directly sampled did not explain attraction (P = 0.2352). However, yeasts isolated from fruit associated niches were more attractive than those from non-fruit associated niches (P = 0.0188) regardless of taxonomic positioning. These data suggest that while attractiveness is primarily correlated with phylogenetic status, the ability to attract Drosophila is a labile trait among yeasts that is potentially associated with those inhabiting fruit ecosystems. Preliminary analysis of the volatiles emitted by four yeast isolates in grape juice show the presence/absence of ethanol and acetic acid were not likely explanations for the observed variation in attraction. These data demonstrate variation among yeasts for their ability to attract Drosophila in a pattern that is consistent with the hypothesis that certain yeasts are manipulating fruit odours to mediate interactions with their Drosophila dispersal agent. © 2013 Palanca et al

Dialectical relations between standard Building Information Modeling tools and specific user requirements

Thesis (S.M. in Architecture Studies)--Massachusetts Institute of Technology, Dept. of Architecture, 2012.This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections.Cataloged from student submitted PDF version of thesis.Includes bibliographical references (p. 144-147).The use of Building Information Modeling (BIM) tools is increasing across the Architectural, Engineering and Construction (AEC) industry. This technology is being adopted in many different countries, in a wide range of types of projects, and by professionals from different disciplines. In other words, BIM tools are being applied in many different contexts of use. Consequently, the requirements of its users are becoming heterogeneous and this heterogeneity hinders the development of BIM tools that can satisfy all possible user requirements. Instead, tools are developed to satisfy more broad, general, and generic needs of the AEC industry. The present thesis examines how BIM users are adapting the standard tools to satisfy their specific requirements. Utilizing the user innovation theory as the framework of analysis, the thesis examines whether and how BIM users are adapting the tools to respond to their requirements through user innovation. Studying eight specific BIM user innovation cases, from different contexts of use, the thesis presents and analyzes the processes underlying BIM user innovation, from the starting motivation, to the final distribution of the actual innovation. This analysis has two main objectives: first, to recognize whether there is user innovation in the case of BIM tools; and second, to understand how that innovation is developed. Finally, the thesis extracts patterns of innovation, and examines whether these user innovation cases fit the model described by the user innovation theory.by Carolina M. Soto Ogueta.S.M.in Architecture Studie