Top-Down Mass Analysis of Protein Tyrosine Nitration: Comparison of Electron Capture Dissociation with “Slow-Heating” Tandem Mass Spectrometry Methods

Tyrosine nitration in proteins is an important post-translational modification (PTM) linked to various pathological conditions. When multiple potential sites of nitration exist, tandem mass spectrometry (MS/MS) methods provide unique tools to locate the nitro-tyrosine(s) precisely. Electron capture dissociation (ECD) is a powerful MS/MS method, different in its mechanisms to the “slow-heating” threshold fragmentation methods, such as collision-induced dissociation (CID) and infrared multiphoton dissociation (IRMPD). Generally, ECD provides more homogeneous cleavage of the protein backbone and preserves labile PTMs. However recent studies in our laboratory demonstrated that ECD of doubly charged nitrated peptides is inhibited by the large electron affinity of the nitro group, while CID efficiency remains unaffected by nitration. Here, we have investigated the efficiency of ECD versus CID and IRMPD for top-down MS/MS analysis of multiply charged intact nitrated protein ions of myoglobin, lysozyme, and cytochrome c in a commercial Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer. CID and IRMPD produced more cleavages in the vicinity of the sites of nitration than ECD. However the total number of ECD fragments was greater than those from CID or IRMPD, and many ECD fragments contained the site(s) of nitration. We conclude that ECD can be used in the top-down analysis of nitrated proteins, but precise localization of the sites of nitration may require either of the “slow-heating” methods

Diseño de un estudio ergonómico para evaluar patologías osteomusculares en puestos laborales. Estudio piloto

El propósito de este estudio es la posibilidad de diseñar un estudio ergonómico de los diferentes puestos de trabajo de una empresa, atendiendo a los de mayor significación en cuanto a producir lesiones osteomusculares en los trabajadores, intentando valorar dichos puestos desde el punto de vista biomecánica, para evaluar el impacto de los movimientos y sobreesfuerzos realizados durante toda la jornada laboral y relacionarlos con los índices de lesiones producidos en dichos trabajadores. Una vez realizado este estudio y si aporta valores significativos, se procederá a valorar las posibles soluciones y mejoras que se puedan implantar, siempre dentro de una viabilidad empresarial y posteriormente ver si dicha implantación ha sido beneficiosa para la salud de los trabajadores

Identifying past social-ecological thresholds to understand long-term temporal dynamics in Spain

A thorough understanding of long-term temporal social-ecological dynamics at the national scale helps to explain the current condition of a country’s ecosystems and to support environmental policies to tackle future sustainability challenges. We aimed to develop a methodological approach to understand past long-term (1960-2010) social-ecological dynamics in Spain. First, we developed a methodical framework that allowed us to explore complex social-ecological dynamics among biodiversity, ecosystem services, human well-being, drivers of change, and institutional responses. Second, we compiled 21 long-term, national-scale indicators and analyzed their temporal relationships through a redundancy analysis. Third, we used a Bayesian change point analysis to detect evidence of past social-ecological thresholds and historical time periods. Our results revealed that Spain has passed through four socialecological thresholds that define five different time periods of nature and society relationships. Finally, we discussed how the proposed methodological approach helps to reinterpret national-level ecosystem indicators through a new conceptual lens to develop a more systems-based way of understanding long-term social-ecological patterns and dynamicsThis work was supported by the Biodiversity Foundation (http://www.fundacion-biodiversidad.es/) of the Spanish Ministry of Agriculture, Food and Environment. Partial financial support was also provided by the Ministry of Economy and Competitiveness of Spain (project CGL2014-53782-P: ECOGRADIENTES). The Spanish National Institute for Agriculture and Food Research and Technology (INIA) funded Marina García-Llorente as part of the European Social Fund. Blanca González García-Mon participated in this article as a “la Caixa” Banking Foundation scholar. The funders had no role in the study design, data collection and analysis, preparation of the report, or the decision to submit the study for publicatio

Fully resolved quiet-Sun magnetic flux tube observed with the Sunrise IMaX instrument

Until today, the small size of magnetic elements in quiet Sun areas has required the application of indirect methods, such as the line-ratio technique or multi-component inversions, to infer their physical properties. A consistent match to the observed Stokes profiles could only be obtained by introducing a magnetic filling factor that specifies the fraction of the observed pixel filled with magnetic field. Here, we investigate the properties of a small magnetic patch in the quiet Sun observed with the IMaX magnetograph on board the balloon-borne telescope Sunrise with unprecedented spatial resolution and low instrumental stray light. We apply an inversion technique based on the numerical solution of the radiative transfer equation to retrieve the temperature stratification and the field strength in the magnetic patch. The observations can be well reproduced with a one-component, fully magnetized atmosphere with a field strength exceeding 1 kG and a significantly enhanced temperature in the mid- to upper photosphere with respect to its surroundings, consistent with semi-empirical flux tube models for plage regions. We therefore conclude that, within the framework of a simple atmospheric model, the IMaX measurements resolve the observed quiet-Sun flux tube.Comment: Accepted for publication in The Astrophysical Journal Letters on Aug 11 201

Detection of vortex tubes in solar granulation from observations with Sunrise

We have investigated a time series of continuum intensity maps and corresponding Dopplergrams of granulation in a very quiet solar region at the disk center, recorded with the Imaging Magnetograph eXperiment (IMaX) on board the balloon-borne solar observatory Sunrise. We find that granules frequently show substructure in the form of lanes composed of a leading bright rim and a trailing dark edge, which move together from the boundary of a granule into the granule itself. We find strikingly similar events in synthesized intensity maps from an ab initio numerical simulation of solar surface convection. From cross sections through the computational domain of the simulation, we conclude that these `granular lanes' are the visible signature of (horizontally oriented) vortex tubes. The characteristic optical appearance of vortex tubes at the solar surface is explained. We propose that the observed vortex tubes may represent only the large-scale end of a hierarchy of vortex tubes existing near the solar surface.Comment: Astrophysical Journal Letters: Sunrise Special Issue, reveived 2010 June 16; accepted 2010 August

Moving Magnetic Features around a Pore

Spectropolarimetric observations from Sunrise II/IMaX obtained in June 2013 are used for a statistical analysis to determine the physical properties of moving magnetic features (MMFs) observed near a pore. MMFs of the same and opposite polarity with respect to the pore are found to stream from its border at an average speed of 1.3 km s$^{-1}$ and 1.2 km s$^{-1}$ respectively, with mainly same-polarity MMFs found further away from the pore. MMFs of both polarities are found to harbor rather weak, inclined magnetic fields. Opposite-polarity MMFs are blue-shifted, while same-polarity MMFs do not show any preference for up- or downflows. Most of the MMFs are found to be of sub-arcsecond size and carry a mean flux of $\sim$ 1.2$\times 10^{17}$ Mx.Comment: 8 pages, 4 figures, accepted for publication in ApJ

Applicability of Milne-Eddington inversions to high spatial resolution observations of the quiet Sun

The physical conditions of the solar photosphere change on very small spatial scales both horizontally and vertically. Such a complexity may pose a serious obstacle to the accurate determination of solar magnetic fields. We examine the applicability of Milne-Eddington (ME) inversions to high spatial resolution observations of the quiet Sun. Our aim is to understand the connection between the ME inferences and the actual stratifications of the atmospheric parameters. We use magnetoconvection simulations of the solar surface to synthesize asymmetric Stokes profiles such as those observed in the quiet Sun. We then invert the profiles with the ME approximation. We perform an empirical analysis of the heights of formation of ME measurements and analyze the uncertainties brought about by the ME approximation. We also investigate the quality of the fits and their relationship with the model stratifications. The atmospheric parameters derived from ME inversions of high-spatial resolution profiles are reasonably accurate and can be used for statistical analyses of solar magnetic fields, even if the fit is not always good. We also show that the ME inferences cannot be assigned to a specific atmospheric layer: different parameters sample different ranges of optical depths, and even the same parameter may trace different layers depending on the physical conditions of the atmosphere. Despite this variability, ME inversions tend to probe deeper layers in granules as compared with intergranular lanes.Comment: Accepted for publication in Astronomy and Astrophysic

Caulobacter crescentus CdnL is a non-essential RNA polymerase-binding protein whose depletion impairs normal growth and rRNA transcription.

CdnL is an essential RNA polymerase (RNAP)-binding activator of rRNA transcription in mycobacteria and myxobacteria but reportedly not in Bacillus. Whether its function and mode of action are conserved in other bacteria thus remains unclear. Because virtually all alphaproteobacteria have a CdnL homolog and none of these have been characterized, we studied the homolog (CdnL <sub>Cc</sub> ) of the model alphaproteobacterium Caulobacter crescentus. We show that CdnL <sub>Cc</sub> is not essential for viability but that its absence or depletion causes slow growth and cell filamentation. CdnL <sub>Cc</sub> is degraded in vivo in a manner dependent on its C-terminus, yet excess CdnL <sub>Cc</sub> resulting from its stabilization did not adversely affect growth. We find that CdnL <sub>Cc</sub> interacts with itself and with the RNAP β subunit, and localizes to at least one rRNA promoter in vivo, whose activity diminishes upon depletion of CdnL <sub>Cc</sub> . Interestingly, cells expressing CdnL <sub>Cc</sub> mutants unable to interact with the RNAP were cold-sensitive, suggesting that CdnL <sub>Cc</sub> interaction with RNAP is especially required at lower than standard growth temperatures in C. crescentus. Our study indicates that despite limited sequence similarities and regulatory differences compared to its myco/myxobacterial homologs, CdnL <sub>Cc</sub> may share similar biological functions, since it affects rRNA synthesis, probably by stabilizing open promoter-RNAP complexes