On the origin of the selectivity of plasmidic H-NS towards horizontally acquired DNA: Linking H-NS oligomerization and cooperative DNA binding

The nucleoid-associated protein H-NS is a global modulator of the expression of genes associated with adaptation to environmental changes. A variant of H-NS expressed in the R27 plasmid was previously shown to selectively modulate the expression of horizontally acquired genes, with minimal effects on core genes that are repressed by the chromosomal form of H-NS. Both H-NS proteins are formed by an oligomerization domain and a DNA-binding domain, which are connected by a linker that is highly flexible in the absence of DNA. We studied DNA binding by means of oligomer-forming chimeric proteins in which domains of the chromosomal and plasmidic variants are exchanged, as well as in monomeric truncated forms containing the DNA-binding domain and variable portions of the linker. Point mutations in the linker were also examined in full-length and truncated H-NS constructs. These experiments show that the linker region contributes to DNA binding affinity and that it is a main component of the distinct DNA binding properties of chromosomal and plasmidic H-NS. We propose that interactions between the linker and DNA limit the flexibility of the connection between H- NS oligomerization and DNA binding and provide an allosteric indirect readout mechanism to detect long- range distortions of DNA, thus enabling discrimination between core and horizontally acquired DNA.

Protein loop compaction and the origin of the effect of arginine and glutamic acid mixtures on solubility, stability and transient oligomerization of proteins

Addition of a 50 mM mixture of l-arginine and l-glutamic acid (RE) is extensively used to improve protein solubility and stability, although the origin of the effect is not well understood. We present Small Angle X-ray Scattering (SAXS) and Nuclear Magnetic Resonance (NMR) results showing that RE induces protein compaction by collapsing flexible loops on the protein core. This is suggested to be a general mechanism preventing aggregation and improving resistance to proteases and to originate from the polyelectrolyte nature of RE. Molecular polyelectrolyte mixtures are expected to display long range correlation effects according to dressed interaction site theory. We hypothesize that perturbation of the RE solution by dissolved proteins is proportional to the volume occupied by the protein. As a consequence, loop collapse, minimizing the effective protein volume, is favored in the presence of RE.

Determination and characterization of quinolones in foodstuffs of animal origin by CE-UV, LC-UV, LC-Fl, LC-MS and LC-MS/MS

In this work methods for the multiresidue determination of the series of quinolones include in the European regulation in food of animal origin are de veloped and validated in line with Commission Decision 2002/657/EC in terms of linearity, decision limit, capability detection, precision and stability. Mult iresidue methods were established to allow the determination of quinolones covered by EU legislation in 2377/90/EC in muscle of chicken, turkey, pig and cow, plasma of cow and pig, liver of pig and milk of cow. First an extraction step was optimized and a SPE step was applied to clean!up and preconcentrate quinolones prior to their separation by CE or LC and determination by CE!UV, LC!UV, LC!Fl, LC!MS with different ion sources (ESI ,ApCI) and different mass analyser (Q, ToF) and LC!E SI!QqQ tandem mass spectrometry. The limits of quantification obtained are always lower than Maxim um Residue Limit (MRL) established by EU for quinolones in animal products and they can be applied to the control of quinolones in foodstuffs of animal origin . Finally the proposed methods were applied to determine quinolones in samples of turkey and pig muscle, pig plasma and milk of cow. Excellent quality parameters and reduced time of analysis were obtained when LC!ESI!MS/MS is used, although the others techniques presented too satisfactory results.

Orofacial pain of cardiac origin: review literature and clinical cases

The most common types of orofacial pain originate at the dental or periodontal level or in the musculoskeletal structures. However, the patient may present pain in this region even though the source is located elsewhere in the body. One possible source of heterotopic pain is of cardiac origin. Objectives: Report two cases of orofacial pain of cardiac origin and review the clinical cases described in the literature. Study Design: Description of clinical cases and review of clinical cases. Results and conclusions: Nine cases of atypical pain of cardiac origin are recorded, which include 5 females and 4 males. In craniofacial structures, pain of cardiac origin is usually bilateral. At the craniofacial level, the most frequent location described is in the throat and jaw. Pain of cardiac origin is considered atypical due to its location, although roughly 10% of the cases of cardiac ischemia manifest primarily in craniofacial structures. Finally, the differential diagnosis of pain of odontogenic origin must be taken into account with pain of non-odontogenic origin (muscle, psychogenic, neuronal, cardiac, sinus and neurovascular pain) in order to avoid diagnostic errors in the dental practice as well as unnecessary treatments.

Comparative genomics of Hedgehog loci in chordates and the origin of Shh regulatory novelties

The origin and evolution of the complex regulatory landscapes of some vertebrate developmental genes, often spanning hundreds of Kbp and including neighboring genes, remain poorly understood. The Sonic Hedgehog (Shh) genomic regulatory block (GRB) is one of the best functionally characterized examples, with several discrete enhancers reported within its introns, vast upstream gene-free region and neighboring genes (Lmbr1 and Rnf32). To investigate the origin and evolution of this GRB, we sequenced and characterized the Hedgehog (Hh) loci from three invertebrate chordate amphioxus species, which share several early expression domains with Shh. Using phylogenetic footprinting within and between chordate lineages, and reporter assays in zebrafish probing >30 Kbp of amphioxus Hh, we report large sequence and functional divergence between both groups. In addition, we show that the linkage of Shh to Lmbr1 and Rnf32, necessary for the unique gnatostomate-specific Shh limb expression, is a vertebrate novelty occurred between the two whole-genome duplications.

Orofacial pain of cardiac origin, serial of clinical cases

Objective: To determine the clinical characteristics of the orofacial pain of cardiac origin in patients visited when doing a treadmill exercise test, at the cardiology service of the Can Ruti Hospital in Badalona (Barcelona, Spain). Study design: The sample of that study included thirty patients visiteding when doing a treadmill exercise test, at the cardiology service. The questionnaire has been asked to a sample of 30 patients. Results: Eleven of the 30 patients included in this study presented craniofacial pain before or during the cardiac seizure. The location of the pain was bilateral, non-irradiated at the mandible in all cases. The intensity of the pain was from slight to severe. The frequency of the appearance of the pain was paroxysmal in 8 cases and constant in three cases, and the duration was from a few hours to a maximum of 14 days. Discussion: The cardiac pain in craniofacial structures is usually bilateral, compared to odontogenic pain which is always unilateral. The pain of cardiac origin is considered atypical because of its location, but about the 10 % of the cases, the cardiac ischemia has its primary manifestation in orofacial structures. Conclusions: Eleven patients referred a bilateral non-irradiated mandibular pain, with intensity from slight to severe, and with a paroxystic frequency in eight cases and a constant frequency in three cases. Just one patient referred pain during the treadmill exercise test. In all cases the pain disappeared after the cardiac surgery or the administration of vasodilators.

Geographic assignment of seabirds to their origin: combining morphology, genetics and biogeochemical analyses

Longline fisheries, oil spills, and offshore wind farms are some of the major threats increasing seabird mortality at sea, but the impact of these threats on specific populations has been difficult to determine so far. We tested the use of molecular markers, morphometric measures, and stable isotope (δ15N and δ13C) and trace element concentrations in the first primary feather (grown at the end of the breeding period) to assign the geographic origin of Calonectris shearwaters. Overall, we sampled birds from three taxa: 13 Mediterranean Cory's Shearwater (Calonectris diomedea diomedea) breeding sites, 10 Atlantic Cory's Shearwater (Calonectris diomedea borealis) breeding sites, and one Cape Verde Shearwater (C. edwardsii) breeding site. Assignment rates were investigated at three spatial scales: breeding colony, breeding archipelago, and taxa levels. Genetic analyses based on the mitochondrial control region (198 birds from 21 breeding colonies) correctly assigned 100% of birds to the three main taxa but failed in detecting geographic structuring at lower scales. Discriminant analyses based on trace elements composition achieved the best rate of correct assignment to colony (77.5%). Body measurements or stable isotopes mainly succeeded in assigning individuals among taxa (87.9% and 89.9%, respectively) but failed at the colony level (27.1% and 38.0%, respectively). Combining all three approaches (morphometrics, isotopes, and trace elements on 186 birds from 15 breeding colonies) substantially improved correct classifications (86.0%, 90.7%, and 100% among colonies, archipelagos, and taxa, respectively). Validations using two independent data sets and jackknife cross-validation confirmed the robustness of the combined approach in the colony assignment (62.5%, 58.8%, and 69.8% for each validation test, respectively). A preliminary application of the discriminant model based on stable isotope δ15N and δ13C values and trace elements (219 birds from 17 breeding sites) showed that 41 Cory's Shearwaters caught by western Mediterranean long-liners came mainly from breeding colonies in Menorca (48.8%), Ibiza (14.6%), and Crete (31.7%). Our findings show that combining analyses of trace elements and stable isotopes on feathers can achieve high rates of correct geographic assignment of birds in the marine environment, opening new prospects for the study of seabird mortality at sea.

Origin of magnetic and magnetoelastic tweedlike precursor modulations in ferroic materials

Based on experimental observations of modulated magnetic patterns in a Co0.5Ni0.205Ga0.295 alloy, we propose a model to describe a (purely) magnetic tweed and a magnetoelastic tweed. The former arises above the Curie (or Nel) temperature due to magnetic disorder. The latter results from compositional fluctuations coupling to strain and then to magnetism through the magnetoelastic interaction above the structural transition temperature. We discuss the origin of purely magnetic and magnetoelastic precursor modulations and their experimental thermodynamic signatures.

Reliable entertainment: Spanish consumers' preferences regarding a film's country of origin

This article contributes to the study of cinema audiences in Europe by analyzing theactual behavior of Spanish moviegoers and their level of satisfaction. We modeledmoviegoers’ choice of film by country of origin (U.S.A., Spain, and other countries)according to a set of determinants: (1) consumers’ interpretation of several sources of information, (2) motivations and (3) choice rules. We found three clear consumerstereotypes related to each type of film: (1) U.S.A. films were preferred by almost everyone (especially families and younger audiences); (2) Spanish films had audiences composed of middle-age and middle-class moviegoers; and (3) European productions were preferred by a social or intellectual elite. U.S.A. films dominate the Spanish market for the reason that they provide most of what moviegoers prefer, namely, familiar, reliable entertainment in Spanish; three characteristics that are not satisfied by Spanish and European films. Additionally, we discuss the implications for the European cultural policy

Tracing the Origin and Spread of Agriculture in Europe

The origins of early farming and its spread to Europe have been the subject of major interest for some time. The main controversy today is over the nature of the Neolithic transition in Europe: the extent to which the spread was, for the most part, indigenous and animated by imitatio (cultural diffusion) or else was driven by an influx of dispersing populations (demic diffusion). We analyze the spatiotemporal dynamics of the transition using radiocarbon dates from 735 early Neolithic sites in Europe, the Near East, and Anatolia. We compute great-circle and shortest-path distances from each site to 35 possible agricultural centers of origin—ten are based on early sites in the Middle East and 25 arehypothetical locations set at 58 latitude/longitude intervals. We perform a linear fit of distance versus age (and viceversa) for each center. For certain centers, high correlation coefficients (R . 0.8) are obtained. This implies that a steady rate or speed is a good overall approximation for this historical development. The average rate of the Neolithic spread over Europe is 0.6–1.3 km/y (95% confidence interval). This is consistent with the prediction of demic diffusion(0.6–1.1 km/y). An interpolative map of correlation coefficients, obtained by using shortest-path distances, shows thatthe origins of agriculture were most likely to have occurred in the northern Levantine/Mesopotamian area