The genomic and physical organization of Ty1-copia-like sequences as a component of large genomes in Pinus elliottii var. elliottii and other gymnosperms.

A DNA sequence, TPE1, representing the internal domain of a Ty1-copia retroelement, was isolated from genomic DNA of Pinus elliottii Engelm. var. elliottii (slash pine). Genomic Southern analysis showed that this sequence, carrying partial reverse transcriptase and integrase gene sequences, is highly amplified within the genome of slash pine and part of a dispersed element >4.8 kbp. Fluorescent in situ hybridization to metaphase chromosomes shows that the element is relatively uniformly dispersed over all 12 chromosome pairs and is highly abundant in the genome. It is largely excluded from centromeric regions and intercalary chromosomal sites representing the 18S-5.8S-25S rRNA genes. Southern hybridization with specific DNA probes for the reverse transcriptase gene shows that TPE1 represents a large subgroup of heterogeneous Ty1-copia retrotransposons in Pinus species. Because no TPE1 transcription could be detected, it is most likely an inactive element--at least in needle tissue. Further evidence for inactivity was found in recombinant reverse transcriptase and integrase sequences. The distribution of TPE1 within different gymnosperms that contain Ty1-copia group retrotransposons, as shown by a PCR assay, was investigated by Southern hybridization. The TPE1 family is highly amplified and conserved in all Pinus species analyzed, showing a similar genomic organization in the three- and five-needle pine species investigated. It is also present in spruce, bald cypress (swamp cypress), and in gingko but in fewer copies and a different genomic organization.

Increased mutation frequency of feline immunodeficiency virus lacking functional deoxyuridine-triphosphatase.

Feline immunodeficiency virus (FIV) encodes the enzyme deoxyuridine-triphosphatase (DU; EC 3.6.1.23) between the coding regions for reverse transcriptase and integrase in the pol gene. Here, we report the in vivo infection of cats with a DU- variant of the PPR strain of FIV and compare its growth properties and tissue distribution with those of wild-type FIV-PPR. The results reveal several important points: (i) DU- FIV is able to infect the cat, with kinetics similar to that observed with wild-type FIV; (ii) both wild-type and DU- FIV-infected specific-pathogen free cats mount a strong humoral antibody response which is able to limit the virus burden in both groups of animals; (iii) the virus burden is reduced in the DU- FIV-infected cats, particularly in tissues such as spleen and salivary gland; and (iv) the mutation frequency in DU- FIVs integrated in the DNA of primary macrophages after 9 months of infection is approximately 5-fold greater than the frequency observed in DU- FIV DNA integrated in T lymphocytes. Mutation rate with wild-type FIV remains the same in both cell types in vivo. The dominant mutations seen in macrophages with DU- FIV are G-->A base changes, consistent with an increased misincorporation of deoxyuridine into viral DNA of DU- FIVs during reverse transcription. Because this enzyme is absent from human immunodeficiency virus type 1 and other primate lentiviruses, virus replication in cell environments with low DU activity may lead to increased mutation and contribute to the rapid expansion of the viral repertoire.

Isolation and characterization of integron-containing bacteria without antibiotic selection

The emergence of antibiotic resistance among pathogenic and commensal bacteria has become a serious problem worldwide. The use and overuse of antibiotics in a number of settings are contributing to the development of antibiotic-resistant microorganisms. The class 1 and 2 integrase genes (intI1 and intI2, respectively) were identified in mixed bacterial cultures enriched from bovine feces by growth in buffered peptone water (BPW) followed by integrase-specific PCR. Integrase-positive bacterial colonies from the enrichment cultures were then isolated by using hydrophobic grid membrane filters and integrase-specific gene probes. Bacterial clones isolated by this technique were then confirmed to carry integrons by further testing by PCR and DNA sequencing. Integron-associated antibiotic resistance genes were detected in bacteria such as Escherichia coli, Aeromonas spp., Proteus spp., Morganella morganii, Shewanella spp., and urea-positive Providencia stuartii isolates from bovine fecal samples without the use of selective enrichment media containing antibiotics. Streptomycin and trimethoprim resistance were commonly associated with integrons. The advantages conferred by this methodology are that a wide variety of integron-containing bacteria may be simultaneously cultured in BPW enrichments and culture biases due to antibiotic selection can be avoided. Rapid and efficient identification, isolation, and characterization of antibiotic resistance-associated integrons are possible by this protocol. These methods will facilitate greater understanding of the factors that contribute to the presence and transfer of integron-associated antibiotic resistance genes in bacterial isolates from red meat production animals.

Active genetic elements present in the locus of enterocyte effacement in Escherichia coli O26 and their role in mobility

The locus of enterocyte effacement (LEE) is a large multigene chromosomal segment encoding gene products responsible for the generation of attaching and effacing lesions in many diarrheagenic Escherichia coli strains. A recently sequenced LEE harboring a pathogenicity island (PAI) from a Shiga toxin E. coli serotype 026 strain revealed a LEE PAI (designated LEE 026) almost identical to that obtained from a rabbit-specific enteropathogenic 015:H- strain. LEE 026 comprises 59,540 bp and is inserted at 94 min within the mature pheU tRNA locus. The LEE 026 PAI is flanked by two direct repeats of 137 and 136 bp (DR1 and DR2), as well as a gene encoding an integrase belonging to the P4 integrase family. We examined LEE 026 for horizontal gene transfer. By generating mini-LEE plasmids harboring only DR1 or DR2 with or without the integrase-like gene, we devised a simple assay to examine recombination processes between these sequences. Recombination was shown to be integrase dependent in a Delta recA E. coli K-12 strain background. Recombinant plasmids harboring a single direct repeat cloned either with or without the LEE 026 integrase gene were found to insert within the chromosomal pheU locus of E. coli K-12 strains with equal efficiency, suggesting that an endogenous P4-like integrase can substitute for this activity. An integrase with strong homology to the LEE 026 integrase was detected on the K-12 chromosome associated with the leuX tRNA locus at 97 min. Strains deleted for this integrase demonstrated a reduction in the insertion frequency of plasmids harboring only the DR into the pheU locus. These results provide strong evidence that LEE-harboring elements are indeed mobile and suggest that closely related integrases present on the chromosome of E. coli strains contribute to the dynamics of PAI mobility.

Circular migration and the Internet: The role of social networks in the sociocultural context of Mexican residents in Barcelona

This work focuses on the study of the circular migration between America and Europe, particularly in the discussion about knowledge transfer and the way that social networks reconfigure the form of information distribution among people, that due to labor and academic issues have left their own country. The main purpose of this work is to study the impact of social media use in migration flows between Mexico and Spain, more specifically the use by Mexican migrants who have moved for  multiple years principally for educational purposes and then have returned to their respective locations in Mexico seeking to integrate themselves into the labor market. Our data collection concentrated exclusively on a group created on Facebook by Mexicans who mostly reside in Barcelona, Spain or wish to travel to the city for economic, educational or tourist reasons.  The results of this research show that while social networks are spaces for exchange and integration, there is a clear tendency by this group to "narrow lines" and to look back to their homeland, slowing the process of opening socially in their new context.

Cross-resistance to elvitegravir and dolutegravir in 502 patients failing on raltegravir: a French national study of raltegravir-experienced HIV-1-infected patients

International audience

Integración de dispositivos electrónicos inteligentes en Smart Grid

El sector eléctrico está experimentando cambios importantes tanto a nivel de gestión como a nivel de mercado. Una de las claves que están acelerando este cambio es la penetración cada vez mayor de los Sistemas de Generación Distribuida (DER), que están dando un mayor protagonismo al usuario a la hora de plantear la gestión del sistema eléctrico. La complejidad del escenario que se prevé en un futuro próximo, exige que los equipos de la red tenga la capacidad de interactuar en un sistema mucho más dinámico que en el presente, donde la interfaz de conexión deberá estar dotada de la inteligencia necesaria y capacidad de comunicación para que todo el sistema pueda ser gestionado en su conjunto de manera eficaz. En la actualidad estamos siendo testigos de la transición desde el modelo de sistema eléctrico tradicional hacia un nuevo sistema, activo e inteligente, que se conoce como Smart Grid. En esta tesis se presenta el estudio de un Dispositivo Electrónico Inteligente (IED) orientado a aportar soluciones para las necesidades que la evolución del sistema eléctrico requiere, que sea capaz de integrase en el equipamiento actual y futuro de la red, aportando funcionalidades y por tanto valor añadido a estos sistemas. Para situar las necesidades de estos IED se ha llevado a cabo un amplio estudio de antecedentes, comenzando por analizar la evolución histórica de estos sistemas, las características de la interconexión eléctrica que han de controlar, las diversas funciones y soluciones que deben aportar, llegando finalmente a una revisión del estado del arte actual. Dentro de estos antecedentes, también se lleva a cabo una revisión normativa, a nivel internacional y nacional, necesaria para situarse desde el punto de vista de los distintos requerimientos que deben cumplir estos dispositivos. A continuación se exponen las especificaciones y consideraciones necesarias para su diseño, así como su arquitectura multifuncional. En este punto del trabajo, se proponen algunos enfoques originales en el diseño, relacionados con la arquitectura del IED y cómo deben sincronizarse los datos, dependiendo de la naturaleza de los eventos y las distintas funcionalidades. El desarrollo del sistema continua con el diseño de los diferentes subsistemas que lo componen, donde se presentan algunos algoritmos novedosos, como el enfoque del sistema anti-islanding con detección múltiple ponderada. Diseñada la arquitectura y funciones del IED, se expone el desarrollo de un prototipo basado en una plataforma hardware. Para ello se analizan los requisitos necesarios que debe tener, y se justifica la elección de una plataforma embebida de altas prestaciones que incluye un procesador y una FPGA. El prototipo desarrollado se somete a un protocolo de pruebas de Clase A, según las normas IEC 61000-4-30 e IEC 62586-2, para comprobar la monitorización de parámetros. También se presentan diversas pruebas en las que se han estimado los retardos implicados en los algoritmos relacionados con las protecciones. Finalmente se comenta un escenario de prueba real, dentro del contexto de un proyecto del Plan Nacional de Investigación, donde este prototipo ha sido integrado en un inversor dotándole de la inteligencia necesaria para un futuro contexto Smart Grid.

Multiresistant Salmonella enterica serovar 4,[5],12:i:- in Europe: a new pandemic strain?

A marked increase in the prevalence of S. enterica serovar 4,[5],12:i:- with resistance to ampicillin, streptomycin, sulphonamides and tetracyclines (R-type ASSuT) has been noted in food-borne infections and in pigs/pig meat in several European countries in the last ten years. One hundred and sixteen strains of S. enterica serovar 4,[5],12:i:- from humans, pigs and pig meat isolated in England and Wales, France, Germany, Italy, Poland, Spain and the Netherlands were further subtyped by phage typing, pulsed-field gel electrophoresis and multilocus variable number tandem repeat analysis to investigate the genetic relationship among strains. PCR was performed to identify the fljB flagellar gene and the genes encoding resistance to ampicillin, streptomycin, sulphonamides and tetracyclines. Class 1 and 2 integrase genes were also sought. Results indicate that genetically related serovar 4,[5],12:i:- strains of definitive phage types DT193 and DT120 with ampicillin, streptomycin, sulphonamide and tetracycline resistance encoded by blaTEM, strA-strB, sul2 and tet(B) have emerged in several European countries, with pigs the likely reservoir of infection. Control measures are urgently needed to reduce spread of infection to humans via the food chain and thereby prevent the possible pandemic spread of serovar 4,[5],12:i:- of R-type ASSuT as occurred with S. Typhimurium DT104 during the 1990s.