Polymorphism in the corticotropin-releasing factor receptor 1 (CRF1-R) gene plays a role in shaping the high anxious phenotype of Marchigian Sardinian alcohol-preferring (msP) rats

Introduction: Marchigian Sardinian alcohol-preferring (msP) rats exhibit innate preference for alcohol along with anxious phenotype. In these animals, two single-nucleotide polymorphisms in position -1,836 and -2,097 from the first start codon of the CRF1-R transcript have been found. Materials and Methods: Here, we examined whether these point mutations account for the heightened anxiety-like behavior and stress responsiveness of msP rats. We rederived the msP rats to obtain two distinct lines carrying the wild-type (GG) and point mutations (AA), respectively. Results: CRF1-R gene expression analysis revealed significant dysregulation of the system in the extended amygdala of AA rats. At the behavioral level, using the elevated plus maze, we found that both AA and GG lines had higher basal anxiety compared to Wistar rats. In the defensive burying test, AA rats showed decreased burying behavior compared to the GG and the unselected Wistar lines. Freezing/immobility did not differ among AA and GG but was higher than that of Wistars. The selective CRF1-R antagonist antalarmin (0, 10, and 20 mg/kg) reduced burying behavior in Wistar animals. However, antalarmin (10 mg/kg) tended to increase rather than reducing this behavior when tested in the msP lines, an effect that appeared more marked in the GG as compared to the AA line. Conclusion: The present data suggest that rats with msP genetic background are more anxious and show different sensitivity to stress and CRF1-R blockade than Wistars. The point mutations occurring in the CRF1-R gene do not seem to influence basal anxiety while they appear to affect active responses to stress

A new serotype 14 variant of the pneumococcal Spain9V-3 international clone detected in the central region of Argentina

The penicillin-resistant Spain9V-3 clone of Streptococcus pneumoniae is widespread and presents different serotype variants originating from recombination of the capsular genes. In this work, the genetic relatedness of 29 invasive pneumococci isolated from the central region of Argentina (Cordoba, Buenos Aires, Santa Fe and La Pampa provinces) was assessed by multilocus sequence typing (MLST). All of the penicillin-non-susceptible isolates studied (21/29) belonged to a serotype 14 variant of the Spain 9V-3 clone. This clone was predominant, suggesting that it was responsible for the penicillin resistance spread in this region. Interestingly, this serotype 14 variant (named Cordoba S14V) could be differentiated from the European one by its pbp1a gene, suggesting a different recombinational replacement of the capsular genes. The putative recombination sites were analysed, resulting in the proximal crossover point being clearly localized in the spr0309 gene, with the distal site restricted to the recU gene, confirming a different recombination event. Analysis of the dexB, cpsB, aliA and pbp1a genes from these strains showed a high similarity with the corresponding genes of the Spain14-5 clone, suggesting that the capsular genes were provided by this international clone. Analysis of the genetic polymorphisms of the pbp1a (nt 1473-1922) and spr0309 (nt 1-790) genes is proposed as an epidemiological tool to help recognize the Cordoba S14V of the Spain9V-3 clone. On the other hand, BOX-repeat-based PCR and MLST analyses of serotype 14 strains revealed a divergent epidemiology of the Cordoba S14V, suggesting a non-recent dissemination in the paediatric population. It is suggested that this molecular epidemiology work will be a reference for monitoring the evolution of S14Vs of Spain9V-3, the emergence of new clones and the impact of pneumococcal vaccination programmes in Argentina.Fil: Albarracín Orio, Andrea Georgina. Universidad Católica de Córdoba. Instituto de Investigaciones en Recursos Naturales y Sustentabilidad José Sanchez Labrador S. J. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones en Recursos Naturales y Sustentabilidad José Sanchez Labrador S. J.; ArgentinaFil: Cortes, Paulo. Hospital Pediátrico del Niño Jesús; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Tregnaghi, Miguel. Centro de Desarrollo de Proyectos Avanzados en Pediatria; ArgentinaFil: Piñas, German Eduardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Echenique, Jose Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Yudowski, Silvia. Centro de Desarrollo de Proyectos Avanzados en Pediatria; ArgentinaFil: Carvajal, Lydia. Centro de Desarrollo de Proyectos Avanzados en Pediatria; ArgentinaFil: Culasso, Catalina. Centro de Desarrollo de Proyectos Avanzados en Pediatria; ArgentinaFil: Nobile, Carmen Beatriz. Centro de Desarrollo de Proyectos Avanzados en Pediatria; ArgentinaFil: Figueroa, Myriam Haydee. Centro de Desarrollo de Proyectos Avanzados en Pediatria; ArgentinaFil: Lopardo, Horacio. Fundacion Hospital de Pediatria Professor Dr. Juan P. Garrahan; ArgentinaFil: Hernández, Claudia. Fundacion Hospital de Pediatria Professor Dr. Juan P. Garrahan; ArgentinaFil: Regueira, Mabel. Instituto Nacional de Enfermedades Infecciosas; Argentin

Room-temperature single-phase Li insertion/extraction in nanoscale LixFePO4

Classical electrodes for Li-ion technology operate by either single-phase or two-phase Li insertion/de-insertion processes, with single-phase mechanisms presenting some intrinsic advantages with respect to various storage applications. We report the feasibility to drive the well-established two-phase room-temperature insertion process in LiFePO4 electrodes into a single-phase one by modifying the material's particle size and ion ordering. Electrodes made of LiFePO4 nanoparticles (40 nm) formed by a low-temperature precipitation process exhibit sloping voltage charge/discharge curves, characteristic of a single-phase behaviour. The presence of defects and cation vacancies, as deduced by chemical/physical analytical techniques, is crucial in accounting for our results. Whereas the interdependency of particle size, composition and structure complicate the theorists' attempts to model phase stability in nanoscale materials, it provides new opportunities for chemists and electrochemists because numerous electrode materials could exhibit a similar behaviour at the nanoscale once their syntheses have been correctly worked out

Resistencia de Shigella spp. a los antimicrobianos en Córdoba, Argentina, durante el período 1990-1997

En este estudio se analiza la evolución de la resistencia a los antimicrobianos en 771 aislados de Shigella spp. obtenidos de un total de 9 195 coprocultivos efectuados entre 1990 y 1997 en un hospital infantil de Córdoba, Argentina. S. flexneri, responsable de 73% de las infecciones por Shigella, fue la especie más resistente. La frecuencia de la multirresistencia de S. flexneri a los tres antibacterianos más utilizados (ampicilina, trimetoprima-sulfametoxazol y cloranfenicol) aumentó de 10% en 1990 a 58% en 1997 (P < 0,001). Cuando se considera separadamente cada uno de ellos, la resistencia a la ampicilina aumentó de 60 a 100% (P < 0,001), la resistencia al cloranfenicol de 13 a 71% (P < 0,001) y la resistencia a la trimetoprima-sulfametoxazol de 79 a 84% (P = 0,22). Para S. sonnei, el aumento de la resistencia a la ampicilina (de 36% en 1990 a 54% en 1997) no fue estadísticamente significativo (P = 0,20), ni tampoco lo fue la disminución de la resistencia a la trimetoprima-sulfametoxazol, que pasó de 82% en 1990 a 55% en 1997 (P = 0,08); solo se detectaron dos aislados resistentes al cloranfenicol, uno en 1995 y otro en 1997, y dos con resistencia múltiple a los tres antibióticos. Consideramos obligatorio realizar pruebas de susceptibilidad en cada aislado clínico de Shigella, ya que permiten detectar cambios en el perfil de la resistencia y, con ello, adecuar el tratamiento empírico

Relationship between dorsal muscle progenitors and adult musculature

Resumen del póster presentado al 2nd Meeting of the Portuguese Society for Developmental Biology, celebrado en Lisboa (Portugal) del 24 al 26 de octubre de 2013.The determination and specification of skeletal muscle in vertebrates is orchestrated by the myogenic regulatory factors (MRFs): Myf5, Mrf4, MyoD and Myogenin. Myf5 is the first to be expressed in the embryo, initiating and co-ordinating the myogenic cascade. In absence of Myf5 progenitors fail to be specified at the correct developmental stage. Activation of MyoD rescues the phenotype and myogenesis progresses. In Myf5/MyoD KO animals rescue does not take place and animals lack all skeletal muscle. We and others have shown that Myf5 transcription is controlled by over 25 regulatory elements, most of them drive expression in a specific spatiotemporal context. The Early Epaxial Enhancer (EEE) operates in the dermomyotomal dorsomedial lip (DML) and is the first enhancer to activate Myf5 expression at E8.5 Although the contribution of the different regulatory elements to the expression pattern is well defined, we still lack an understanding on the contribution of the different subpopulations of muscle progenitor cells to adult musculature. Furthermore, there is still no connection between the spatiotemporal activation of Myf5 and its function within the particular set of myogenic precursors in which it is active. In order to address these questions we are following two strategies: 1/ Generation of transgenic strains to analyse the function of the EEE by means of lineage tracing and cell ablation experiments and 2/ Generation of a new mouse knockout strain in which the EEE has been targeted. Comparison of Evolutionary Conserved Regions between different species shows two highly conserved peaks within the experimentally defined EEE element (EEE1 and EEE2). As these could represent different functions within the enhancer, we have cloned each peak in our standard vector and used to generate transgenic animals. Preliminary data show that EEE2 drives reporter gene expression at E8.5 and is turned off at E9.5, while EEE1 activates gene expression at E9.5. We are now transferring these refined enhancers into cell ablation and lineage tracing constructs. We are in the process of analyzing the data obtained by RNAseq from wild type and Myf5EEE-/EEE- embryos in order to determine the downstream targets of Myf5 in the DML. Crucially, we have crossed this new allele into the MyoD KO and show that in the absence of MyoD rescue, some muscles are lost (or severely reduced), linking for the first time a particular set of progenitors to specific adult muscles.Peer Reviewe

“peut-être on peut improviser un peu” cas de co-construction de références ad hoc en situation interactionnelle de jeu

International audienc

“peut-être on peut improviser un peu” cas de co-construction de références ad hoc en situation interactionnelle de jeu

International audienc

: The emergence of joint construction of reference in a card game situation

International audienceFollowing the methodological approach of multimodal conversation analysis, our study focuses on the emergence of a jointly constructed reference during a card game interaction: the participants need to find a common solution in order to continue their activity. The data collected in situ allow us to apprehend the different verbal and non-verbal resources mobilised by the players in order to show that in social interaction reference construction is a joint achievement that involves various types of resources which are temporally finely tuned (among others joint visual attention on the object gesturally put in focus). We also show that once a referent-function association is established and grounded, it can be "activated" later on by using an iconic gesture

New roles of MYF5 in dorsal somitic progenitors

Resumen del trabajo presentado en el European Developmental Biology Congress, celebrado en Alicante (España), del 23 al 26 de octubre de 2019Skeletal muscles originate from different mesoderms: presomitic (limb and trunk muscles), head (extraocular muscles) and pharyngeal mesoderm (facial muscles). Independently from their origin, all muscles develop under the control of the four Myogenic Regulatory Factors: Myf5, Mrf4, MyoD and MyoG. Myf5 is the first to be expressed, and controls myogenic specification. MyoD function overlaps with that of Myf5, and rescues the Myf5-KO phenotype. Myf5/MyoD double mutants have a general lack of skeletal muscles as myogenic precursors are not specified. MyoG drives terminal differentiation and in its absence adult muscles are not formed. Mrf4 plays roles in specification and differentiation, although its function remains poorly understood. The Early Epaxial Enhancer (EEE) directs the earliest Myf5 expression, starting in the somitic dorsomedial lip. We have generated a new mouse allele in which the EEE has been removed. This new mutant only loses Myf5 expression in the dorsal part of the first 5-6 somites. RNA-seq reveals differences in myogenic, innervation and limb, neurogenesis and chondrogenesis gene-networks. We have now validated several of the identified genes by qPCR and/or ISH. Because EEE-KO animals lack an overt phenotype, we generated EEE/MyoD double mutants, thus abolishing MyoD rescue in dorsal somitic progenitors. In these animals, we observe severe defects in diaphragm, ribcage and posture (kyphosis), which lead us to study the epaxial musculature more in detail to see if there is a particular group of muscles affected. Also, we are trying to elucidate how absence of Myf5 impacts upon Pax1 expression, presumably causing the ribcage defects