Isolation and characterization of ten microsatellite loci from the Patagonian notothenioid fish Eleginops maclovinus

Eleginops maclovinus, the Patagonian blennie, is a notothenioid (Perciformes) endemic to South American temperate and sub-Antarctic waters. Here, we report ten polymorphic microsatellite loci isolated from a dinucleotide-enriched E. maclovinus genomic library. Among 48 individuals, the number of alleles per locus ranged from eight to 41, and the observed and expected heterozygosity ranged from 0.688 to 0.938 and from 0.695 to 0.968, respectively. No locus significantly deviated from Hardy-Weinberg equilibrium, and no significant linkage disequilibrium between pairs of loci was found. These polymorphic microsatellite loci will be useful for investigating genetic population structure and connectivity among natural populations.

Observed supersymmetry of hadrons and the tachyonless string models

The experimental mesonic density of states ρmeson(m)≃ρbaryon(m) from 0.9 to 1.3 GeV. In this region the ρmeson fits the ρ(m) deduced for it from discrete bag model states. Beyond 1.3 GeV one can expect exotic mesons. If ρmeson is replaced by the baryon density (as suggested by string model studies [D. Kutasov and N. Seiberg, Nucl. Phys. B 358 (1991) 600; P.G.O. Freund and J.L. Rosner, Phys. Rev. Lett. 68 (1992) 765]), agreement with theory is obtained up to 1.7 GeV. Beyond 1.7 GeV exotic baryons may be expected.

Isolation and characterization of microsatellite loci for the mud-dauber wasp Trypoxylon (Trypargilum) albitarse (Hymenoptera: Crabronidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Development and characterization of microsatellite loci of microglanis cottoides (siluriformes: pseudopimelodidae) and cross-species amplification

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Development and characterization of microsatellite markers for Vriesea simplex (Bromeliaceae) and cross-amplification in other species of Bromeliaceae

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

New loci of Lychnophora ericoides and transferability to Lychnophora pinaster, endangered medicinal species from Brazil

Lychnophora ericoides and Lychnophora pinaster are species used in popular medicine as analgesic or anti-inflammatory agents to treat contusions, rheumatism, and insect bites. In this study, 21 simple sequence repeat loci of L. ericoides were developed and transferred to L. pinaster. Three populations of L. ericoides and 2 populations of L. pinaster were evaluated; they were collected in the State of Minas Gerais. Population parameters were estimated, and the mean values of observed and expected heterozygosity were 0.297 and 0.408 (L. ericoides) and 0.228 and 0.310 (L. pinaster), respectively. Greater genetic variability was observed within populations than between populations of L. ericoides (62 and 37%) and L. pinaster (97 and 2.8%). These results provide information for genetic conservation and taxonomic studies of these endangered species.

Geoffroy's side-necked turtle [Phrynops geoffroanus (Schweigger, 1812), Testudines: Chelidae] as a model for evolutionary ecotoxicology: relationship between environmental contamination, conditions and genetic variability

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Genetic variability and differentiation in the Mediterranean endemic starry ray Raja asterias (Delaroche, 1809)

The present study deal with the population structure and connectivity of the Mediterranean endemic starry ray Raja asterias (Delaroche, 1809) in the Western and Eastern Mediterranean basin. A panel of eight microsatellite loci which cross-amplify in Rajidae (El Nagar, 2010) was used to assess population connectivity and structure. Those aims were investigated by analyzing the genetic variation of 9 population sample for a total of 185 individuals collected during past scientific surveys (MEDITS, GRUND), commercial trawling and also directly at fish markets. The purpose of this thesis is to estimate the genetic divergence occurring between the Mediterranean populations and, in particular, to assess the presence of any barrier (geographic, hydrogeological and biological) to gene flow for this species. Different statistical approaches were performed to reach this aim evaluating both the genetic diversity (nucleotide diversity, allelic richness, observed and expected heterozygosity and Hardy-Weinberg equilibrium test) and the population differentiation patterns (pairwise Fst estimated and population structure analysis). The results obtained from the analysis of the microsatellite dataset suggest a geographic and genetic separation between the starry ray populations of the Mediterranean basin into three or four distinct groups: Western and Eastern Mediterranean basins and Sicilian coast always clustering as an independent group and Algeria which could be or not considered another separate group. The data were discussed from both an evolutionary and a conservation point of view and in relation to previous results obtained by the analysis of mitochondrial marker. A comparison with other Mediterranean demersal skate species was performed in order to better contextualise our results. Finally, our results could offer useful information to protect vulnerable species as R. asterias and developing effective conservation plans in the Mediterranean.

Genetic and morphological variation and differentiation of Raja clavata populations in the European seas: marker development and preliminary data

In this study the population structure and connectivity of the Mediterranean and Atlantic Raja clavata (L., 1758) were investigated by analyzing the genetic variation of six population samples (N = 144) at seven nuclear microsatellite loci. The genetic dataset was generated by selecting population samples available in the tissue databases of the GenoDREAM laboratory (University of Bologna) and of the Department of Life Sciences and Environment (University of Cagliari), all collected during past scientific surveys (MEDITS, GRUND) from different geographical locations in the Mediterranean basin and North-east Atlantic sea, as North Sea, Sardinian coasts, Tuscany coasts and Cyprus Island. This thesis deals with to estimate the genetic diversity and differentiation among 6 geographical samples, in particular, to assess the presence of any barrier (geographic, hydrogeological or biological) to gene flow evaluating both the genetic diversity (nucleotide diversity, observed and expected heterozygosity, Hardy- Weinberg equilibrium analysis) and population differentiation (Fst estimates, population structure analysis). In addition to molecular analysis, quantitative representation and statistical analysis of morphological individuals shape are performed using geometric morphometrics methods and statistical tests. Geometric coordinates call landmarks are fixed in 158 individuals belonging to two population samples of Raja clavata and in population samples of closely related species, Raja straeleni (cryptic sibling) and Raja asterias, to assess significant morphological differences at multiple taxonomic levels. The results obtained from the analysis of the microsatellite dataset suggested a geographic and genetic separation between populations from Central-Western and Eastern Mediterranean basins. Furthermore, the analysis also showed that there was no separation between geographic samples from North Atlantic Ocean and central-Western Mediterranean, grouping them to a panmictic population. The Landmark-based geometric morphometry method results showed significant differences of body shape able to discriminate taxa at tested levels (from species to populations).

Unique mixed phenotype and unexpected functional effect revealed by novel compound heterozygosity mutations involving SCN5A.

BACKGROUND Functional characterization of mutations involving the SCN5A-encoded cardiac sodium channel has established the pathogenic mechanisms for type 3 long QT syndrome and type 1 Brugada syndrome and has provided key insights into the physiological importance of essential structure-function domains. OBJECTIVE This study sought to present the clinical and biophysical phenotypes discerned from compound heterozygosity mutations in SCN5A on different alleles in a toddler diagnosed with QT prolongation and fever-induced ventricular arrhythmias. METHODS A 22-month-old boy presented emergently with fever and refractory ventricular tachycardia. Despite restoration of sinus rhythm, the infant sustained profound neurological injury and died. Using polymerase chain reaction, denaturing high-performance liquid chromatography, and direct DNA sequencing, comprehensive open-reading frame/splice mutational analysis of the 12 known long QT syndrome susceptibility genes was performed. RESULTS The infant had 2 SCN5A mutations: a maternally inherited N-terminal frame shift/deletion (R34fs/60) and a paternally inherited missense mutation, R1195H. The mutations were engineered by site-directed mutagenesis and heterologously expressed transiently in HEK293 cells. As expected, the frame-shifted and prematurely truncated peptide, SCN5A-R34fs/60, showed no current. SCN5A-R1195H had normal peak and late current but abnormal voltage-dependent gating parameters. Surprisingly, co-expression of SCN5A-R34fs/60 with SCN5A-R1195H elicited a significant increase in late sodium current, whereas co-expression of SCN5A-WT with SCN5A-R34fs/60 did not. CONCLUSIONS A severe clinical phenotype characterized by fever-induced monomorphic ventricular tachycardia and QT interval prolongation emerged in a toddler with compound heterozygosity involving SCN5A: R34fs/60, and R1195H. Unexpectedly, the 94-amino-acid fusion peptide derived from the R34fs/60 mutation accentuated the late sodium current of R1195H-containing Na(V)1.5 channels in vitro.

Echinococcus P29 antigen: molecular characterization and implication on post-surgery follow-up of CE patients infected with different species of the Echinococcus granulosus complex.

The protein P29 is a potential serological marker for post-treatment monitoring of cystic echinococcosis (CE) especially in young patients. We now have demonstrated that P29 is encoded in the Echinococcus genus by a single gene consisting of 7 exons spanning 1.2 kb of DNA. Variability of the p29 gene at inter- and intra-species level was assessed with 50 cDNA and 280 genomic DNA clones isolated from different E. granulosus s.l. isolates (E. granulosus sensu stricto (G1), E. equinus (G4), E. ortleppi (G5), E. canadensis (G6), E. canadensis (G7) and E. canadensis (G10)) as well as four E. multilocularis isolates. Scarce interspecies polymorphism at the p29 locus was observed and affected predominantly E. granulosus s.s. (G1), where we identified two alleles (A1 and A2) coding for identical P29 proteins and yielding in three genotypes (A1/A1, A2/A2 and A1/A2). Genotypic frequencies expected under Hardy-Weinberg equilibrium revealed a high rate of heterozygosity (47%) that strongly supports the hypothesis that E. granulosus s.s. (G1) is predominantly outbreeding. Comparative sequence analyses of the complete p29 gene showed that phylogenetic relationships within the genus Echinococcus were in agreement with those of previous nuclear gene studies. At the protein level, the deduced P29 amino acid (AA) sequences exhibited a high level of conservation, ranging from 97.9% AA sequence identity among the whole E. granulosus s.l. group to 99.58% identity among E. multilocularis isolates. We showed that P29 proteins of these two species differ by three AA substitutions without implication for antigenicity. In Western-blot analyses, serum antibodies from a human CE patient infected with E. canadensis (G6) strongly reacted with recombinant P29 from E. granulosus s.s. (G1) (recEg(G1)P29). In the same line, human anti-Eg(G1)P29 antibodies bound to recEcnd(G6)P29. Thus, minor AA sequence variations appear not to impair the prognostic serological use of P29.

67P/Churyumov-Gerasimenko: Activity between March and June 2014 as observed from Rosetta/OSIRIS

Aims. 67P/Churyumov-Gerasimenko is the target comet of the ESA’s Rosetta mission. After commissioning at the end of March 2014, the Optical, Spectroscopic, and Infrared Remote Imaging System (OSIRIS) onboard Rosetta, started imaging the comet and its dust environment to investigate how they change and evolve while approaching the Sun. Methods. We focused our work on Narrow Angle Camera (NAC) orange images and Wide Angle Camera (WAC) red and visible-610 images acquired between 2014 March 23 and June 24 when the nucleus of 67P was unresolved and moving from approximately 4.3 AU to 3.8 AU inbound. During this period the 67P – Rosetta distance decreased from 5 million to 120 thousand km. Results. Through aperture photometry, we investigated how the comet brightness varies with heliocentric distance. 67P was likely already weakly active at the end of March 2014, with excess flux above that expected for the nucleus. The comet’s brightness was mostly constant during the three months of approach observations, apart from one outburst that occurred around April 30 and a second increase in flux after June 20. Coma was resolved in the profiles from mid-April. Analysis of the coma morphology suggests that most of the activity comes from a source towards the celestial north pole of the comet, but the outburst that occurred on April 30 released material in a different direction.

Assessment of expected damage on buildings subjected to Lorca earthquake through an energy-based seismic index method and nonlinear dynamic response analyses

The city of Lorca (Spain) was hit on May 11th, 2011, by two consecutive earth-quakes of magnitudes 4.6 and 5.2 Mw, causing casualties and important damage in buildings. Many of the damaged structures were reinforced concrete frames with wide beams. This study quantifies the expected level of damage on this structural type in the case of the Lorca earth-quake by means of a seismic index Iv that compares the energy input by the earthquake with the energy absorption/dissipation capacity of the structure. The prototype frames investigated represent structures designed in two time periods (1994–2002 and 2003–2008), in which the applicable codes were different. The influence of the masonry infill walls and the proneness of the frames to concentrate damage in a given story were further investigated through nonlinear dynamic response analyses. It is found that (1) the seismic index method predicts levels of damage that range from moderate/severe to complete collapse; this prediction is consistent with the observed damage; (2) the presence of masonry infill walls makes the structure very prone to damage concentration and reduces the overall seismic capacity of the building; and (3) a proper hierarchy of strength between beams and columns that guarantees the formation of a strong column-weak beam mechanism (as prescribed by seismic codes), as well as the adoption of counter-measures to avoid the negative interaction between non-structural infill walls and the main frame, would have reduced the level of damage from Iv=1 (collapse) to about Iv=0.5 (moderate/severe damage)

Brown trout thermal niche and climate change: expected changes in the distribution of cold-water fish in central Spain

This paper addresses the determination of the realized thermal niche and the effects of climate change on the range distribution of two brown trout populations inhabiting two streams in the Duero River basin (Iberian Peninsula) at the edge of the natural distribution area of this species. For reaching these goals, new methodological developments were applied to improve reliability of forecasts. Water temperature data were collected using 11 thermographs located along the altitudinal gradient, and they were used to model the relationship between stream temperature and air temperature along the river continuum. Trout abundance was studied using electrofishing at 37 sites to determine the current distribution. The RCP4.5 and RCP8.5 change scenarios adopted by the International Panel of Climate Change for its Fifth Assessment Report were used for simulations and local downscaling in this study. We found more reliable results using the daily mean stream temperature than maximum daily temperature and their respective seven days moving-average to determine the distribution thresholds. Thereby, the observed limits of the summer distribution of brown trout were linked to thresholds between 18.1ºC and 18.7ºC. These temperatures characterise a realised thermal niche narrower than the physiological thermal range. In the most unfavourable climate change scenario, the thermal habitat loss of brown trout increased to 38% (Cega stream) and 11% (Pirón stream) in the upstream direction at the end of the century; however, at the Cega stream, the range reduction could reach 56% due to the effect of a ?warm-window? opening in the piedmont reach.

A resistant genetic background leading to incomplete penetrance of intestinal neoplasia and reduced loss of heterozygosity in ApcMin/+ mice

Previous studies of Min/+ (multiple intestinal neoplasia) mice on a sensitive genetic background, C57BL/6 (B6), showed that adenomas have lost heterozygosity for the germ-line ApcMin mutation in the Apc (adenomatous polyposis coli) gene. We now report that on a strongly resistant genetic background, AKR/J (AKR), Min-induced adenoma multiplicity is reduced by about two orders of magnitude compared with that observed on the B6 background. Somatic treatment with a strong mutagen increases tumor number in AKR Min/+ mice in an age-dependent manner, similar to results previously reported for B6 Min/+ mice. Immunohistochemical analyses indicate that Apc expression is suppressed in all intestinal tumors from both untreated and treated AKR Min/+ mice. However, the mechanism of Apc inactivation in AKR Min/+ mice often differs from that observed for B6 Min/+ mice. Although loss of heterozygosity is observed in some tumors, a significant percentage of tumors showed neither loss of heterozygosity nor Apc truncation mutations. These results extend our understanding of the effects of genetic background on Min-induced tumorigenesis in several ways. First, the AKR strain carries modifiers of Min in addition to Mom1. This combination of AKR modifiers can almost completely suppress spontaneous intestinal tumorigenesis associated with the Min mutation. Second, even on such a highly resistant genetic background, tumor formation continues to involve an absence of Apc function. The means by which Apc function is inactivated is affected by genetic background. Possible scenarios are discussed.