Heterologous reporter expression in the planarian Schmidtea mediterranea through somatic mRNA transfection

Planarians have long been studied for their regenerative abilities. Moving forward, tools for ectopic expression of non-native proteins will be of substantial value. Using a luminescent reporter to overcome the strong autofluorescence of planarian tissues, we demonstrate heterologous protein expression in planarian cells and live animals. Our approach is based on the introduction of mRNA through several nanotechnological and chemical transfection methods. We improve reporter expression by altering untranslated region (UTR) sequences and codon bias, facilitating the measurement of expression kinetics in both isolated cells and whole planarians using luminescence imaging. We also examine protein expression as a function of variations in the UTRs of delivered mRNA, demonstrating a framework to investigate gene regulation at the post-transcriptional level. Together, these advances expand the toolbox for the mechanistic analysis of planarian biology and establish a foundation for the development and expansion of transgenic techniques in this unique model system

Branching ratio change in K- absorption at rest and the nature of the Lambda(1405)

We investigate in-medium corrections to the branching ratio in K- absorption at rest and their effect on the (positively and negatively) charged pion spectrum. The in-medium corrections are due to Pauli blocking, which arises if the Lambda(1405) is assumed to be a $\bar{K}$-nucleon bound state and leads to a density and momentum dependent mass shift of the Lambda(1405). Requiring that the optical potential as well as the branching ratio are derived from the same elementary T-matrix, we find that the in-medium corrected, density dependent T-matrix gives a better description of the K- absorption reaction than the free, density-independent one. This result suggests that the dominant component of the Lambda(1405) wave function is the $\bar{K}N$ bound state.Comment: 8 Pages, Revtex with epsf, and embedded 8 ps figure

The effects of siblings on the migration of women in two rural areas of Belgium and the Netherlands, 1829-1940

This study explores the extent to which the presence and activities of siblings shaped the chances of women migrating to rural and urban areas in two rural areas of Belgium and the Netherlands during the second half of the nineteenth and first decades of the twentieth century. Shared-frailty Cox proportional hazard analyses of longitudinal data from historical population registers show that siblings exerted an additive impact on women's migration, independently of temporal and household characteristics. Just how siblings influenced women's migration depended on regional modes of production and on employment opportunities. In the Zeeland region, sisters channelled each other into service positions. In the Pays de Herve, where men and women found industrial work in the Walloon cities, women were as much influenced by their brothers' activities. Evidence is found for two mechanisms explaining the effects of siblings: micro-economic notions of joint-household decision-making and social capital theory

Position of the Third Na+ Site in the Aspartate Transporter GltPh and the Human Glutamate Transporter, EAAT1

Glutamate transport via the human excitatory amino acid transporters is coupled to the co-transport of three Na+ ions, one H+ and the counter-transport of one K+ ion. Transport by an archaeal homologue of the human glutamate transporters, GltPh, whose three dimensional structure is known is also coupled to three Na+ ions but only two Na+ ion binding sites have been observed in the crystal structure of GltPh. In order to fully utilize the GltPh structure in functional studies of the human glutamate transporters, it is essential to understand the transport mechanism of GltPh and accurately determine the number and location of Na+ ions coupled to transport. Several sites have been proposed for the binding of a third Na+ ion from electrostatic calculations and molecular dynamics simulations. In this study, we have performed detailed free energy simulations for GltPh and reveal a new site for the third Na+ ion involving the side chains of Threonine 92, Serine 93, Asparagine 310, Aspartate 312, and the backbone of Tyrosine 89. We have also studied the transport properties of alanine mutants of the coordinating residues Threonine 92 and Serine 93 in GltPh, and the corresponding residues in a human glutamate transporter, EAAT1. The mutant transporters have reduced affinity for Na+ compared to their wild type counterparts. These results confirm that Threonine 92 and Serine 93 are involved in the coordination of the third Na+ ion in GltPh and EAAT1

Nycthemeral and Monthly Occupation of the Fish Assemblage on a Sheltered Beach of Baía Norte, Florianópolis, Santa Catarina State, Brazil

Interpreting fish community records is challenging for several reasons, including the lack of past ichthyofauna data, the cyclical temporal variations in the community, and the methodology employed, which usually underestimates fish assemblages. The objective of this study was to describe short-scale and meso-scale (nycthemeral period and months, respectively) temporal variations in the ichthyofauna composition and structure of a sheltered beach of Baía Norte (Florianópolis, Santa Catarina state, Brazil), using a capéchade net. Samples were collected monthly for a period of 48 hours. During the period from December 2010 to November 2011, a total of 19,302 individuals belonging to 89 species and 39 families were captured. The number of individuals that were sampled during the day and/or night was dependent on the sampling month. On average, the daytime assemblage was more abundant and different in structure and composition than the nighttime assemblage. Of the eight species that had the highest Index of Relative Importance (%IRI), five had higher variations (ANOVA F) between the day and night than between the months. This finding reinforced the need for sampling during both the day and night. The capéchade net effectively captured demersal and pelagic individuals in a broad range of sizes

Metabolic Profiling of a Mapping Population Exposes New Insights in the Regulation of Seed Metabolism and Seed, Fruit, and Plant Relations

To investigate the regulation of seed metabolism and to estimate the degree of metabolic natural variability, metabolite profiling and network analysis were applied to a collection of 76 different homozygous tomato introgression lines (ILs) grown in the field in two consecutive harvest seasons. Factorial ANOVA confirmed the presence of 30 metabolite quantitative trait loci (mQTL). Amino acid contents displayed a high degree of variability across the population, with similar patterns across the two seasons, while sugars exhibited significant seasonal fluctuations. Upon integration of data for tomato pericarp metabolite profiling, factorial ANOVA identified the main factor for metabolic polymorphism to be the genotypic background rather than the environment or the tissue. Analysis of the coefficient of variance indicated greater phenotypic plasticity in the ILs than in the M82 tomato cultivar. Broad-sense estimate of heritability suggested that the mode of inheritance of metabolite traits in the seed differed from that in the fruit. Correlation-based metabolic network analysis comparing metabolite data for the seed with that for the pericarp showed that the seed network displayed tighter interdependence of metabolic processes than the fruit. Amino acids in the seed metabolic network were shown to play a central hub-like role in the topology of the network, maintaining high interactions with other metabolite categories, i.e., sugars and organic acids. Network analysis identified six exceptionally highly co-regulated amino acids, Gly, Ser, Thr, Ile, Val, and Pro. The strong interdependence of this group was confirmed by the mQTL mapping. Taken together these results (i) reflect the extensive redundancy of the regulation underlying seed metabolism, (ii) demonstrate the tight co-ordination of seed metabolism with respect to fruit metabolism, and (iii) emphasize the centrality of the amino acid module in the seed metabolic network. Finally, the study highlights the added value of integrating metabolic network analysis with mQTL mapping