Structural polymorphism of the major capsid protein of rotavirus

Rotaviruses are important human pathogens with a triple-layered icosahedral capsid. The major capsid protein VP6 is shown here to self-assemble into spherical or helical particles mainly depending upon pH. Assembly is inhibited either by low pH (<3.0) or by a high concentration (>100 mM) of divalent cations (Ca(2+) and Zn(2+)). The structures of two types of helical tubes were determined by electron cryomicroscopy and image analysis to a resolution of 2.0 and 2.5 nm. In both reconstructions, the molecular envelope of VP6 fits the atomic model determined by X-ray crystallography remarkably well. The 3-fold symmetry of the VP6 trimer, being incompatible with the helical symmetry, is broken at the level of the trimer contacts. One type of contact is maintained within all VP6 particles (tubes and virus), strongly suggesting that VP6 assemblies arise from different packings of a unique dimer of trimers. Our data show that the protonation state and thus the charge distribution are important switches governing the assembly of macromolecular assemblies.     

Effect of aproteic diet and fasting on insulin, pancreatic noradrenaline and luteinizing hormone. Changes after 24-hour refeeding

OBJECTIVE AND DESIGN: the objective of this study performed in adult male rats was to determine the alteration in glycemic, insulin and gonadotrophin luteinizing hormone secretion, and noradrenaline pancreatic concentration caused by fasting (F) and aproteic diet (Ap) during 7 and 21 days respectively, as well as the recovery after 24-hour refeeding with control diet (Co). RESULTS: a significant decrease in glycemic levels was only achieved through fasting (F: 86 +/- 5.1 mg %), when compared with controls (Co: 107 +/- 5 mg %). In spite of the high levels of carbohydrates (89%) present in the aproteic diet, the animals fed with this diet showed no differences in glycemic levels (Ap: 120.3 +/- 12.2 mg %), compared with controls. As a result of fasting and aproteic diet, there was a significant decrease in insulin (F: 8.67 +/- 1.36; Ap: 5.7 +/- 0.67; Co: 31 +/- 3.4 uU/ml) and LH levels (F: 10.175 +/- 1.74; Ap: 13.7 +/- 4; Co: 29.83 +/- 4.91 ng/ml). The refed recovered insulin (FR: 50.57 +/- 6.63; ApR: 43.5 +/- 6.85 uU/ml), but not LH levels (FR: 14.25 +/- 3.54; ApR: 13.03 +/- 4.25 ng/ml). A significant increase was observed in the pancreatic noradrenaline concentration (P<0.001) of rats receiving aproteic diet (889.9 +/- 34.65 ng/mg tissue) and fasting during 7 days (827.5 +/- 55.7 ng/mg tissue), compared with controls (531.1 +/- 48.6 ng/mg tissue). CONCLUSIONS: fasting and aproteic diets altered gonadal and metabolic control. When returning to a normal nutritional condition, only the metabolic control, not the reproductive function, could be recovered in the first 24 hours of refeeding. Malnutrition-induced hypoinsulinemia would be caused by an increase in a specific noradrenergic tone.     

Establishment and characterization of a new continuous cell line from Lutzomyia longipalpis (Diptera: psychodidae) and its susceptibility to infections with arboviruses and Leishmania chagasi

Embryonic tissue explants of the sand fly Lutzomyia longipalpis (Lutz & Neiva 1912) the main vector of Leishmania chagasi (Cunha and Chagas), were used to obtain a continuous cell line (Lulo). The tissues were seeded in MM/VP12 medium and these were incubated at 28 masculineC. The first subculture was obtained 45 days after explanting and 96 passages have been made to date. Lulo is composed of epithelioid cells, showed a 0.04 generations/hour exponential growth rate and population doubling time at 24.7 h. The cell line isoenzymatic profiles were determined by using PGI, PGM, MPI and 6-PGDH systems, coinciding with patterns obtained from the same species and colony's pupae and adults. The species karyotype characteristics were recognized (2n = 8), in which pair 1 is subtelocentric and pairs 2, 3 and 4 are metacentric. Lulo was free from bacterial, fungal, mycoplasmic and viral infection. Susceptibility to five arbovirus was determined, the same as Lulo interaction with Leishmania promastigotes.     

Simultaneous determination of albendazole sulfoxide enantiomers and albendazole sulfone in plasma

A high-performance liquid chromatographic method has been developed for the simultaneous determination of albendazole sulfoxide (ABZSO) enantiomers and albendazole sulfone (ABZSO₂) in human plasma. The resolution of ABZSO enantiomers and ABZSO₂ was obtained on a Chiralpak^® AD column using hexane–isopropanol–ethanol (81:14.25:4.75, v/v/v) as the mobile phase. The drugs were detected by fluorescence (λ_exc=280 nm, λ_em=320 nm). The drugs were extracted from 500 μl plasma with ethyl acetate, and after solvent evaporation, the residues were dissolved in the mobile phase and chromatographed. The method was precise and accurate for the three compounds, as judged by the coefficients of variation and relative errors observed. Linear standard curves were obtained in the concentration range of 5–2500 ng/ml for ABZSO enantiomers and 1–500 ng/ml for ABZSO₂. A typical plasma concentration–time profile is presented for one patient under treatment for neurocysticercosis.     

Novel Bread Wheat Lines Enriched in Carotenoids Carrying Hordeum chilense Chromosome Arms in the ph1b Background

The use of crop wild relative species to improve major crops performance is well established. Hordeum chilense has a high potential as a genetic donor to increase the carotenoid content of wheat. Crosses between the 7H^ch H. chilense substitution lines in wheat and the wheat pairing homoeologous1b (ph1b) mutant allowed the development of wheat-H. chilense translocation lines for both 7H^chα and 7H^chβ chromosome arms in the wheat background. These translocation lines were characterized by in situ hybridization and using molecular markers. In addition, reverse phase chromatography (HPLC) analysis was carried out to evaluate the carotenoid content and both 7H^chα∙7AL and 7AS∙7H^chβ disomic translocation lines. The carotenoid content in 7H^chα∙7AL and 7AS∙7H^chβ disomic translocation lines was higher than the wheat-7H^ch addition line and double amount of carotenoids than the wheat itself. A proteomic analysis confirmed that the presence of chromosome 7H^ch introgressions in wheat scarcely altered the proteomic profile of the wheat flour. The Psy1 (Phytoene Synthase1) gene, which is the first committed step in the carotenoid biosynthetic pathway, was also cytogenetically mapped on the 7H^chα chromosome arm. These new wheat-H. chilense translocation lines can be used as a powerful tool in wheat breeding programs to enrich the diet in bioactive compounds.     

RECK-Mediated β1-Integrin Regulation by TGF-β1 Is Critical for Wound Contraction in Mice

Fibroblasts are critical for wound contraction; a pivotal step in wound healing. They produce and modify the extracellular matrix (ECM) required for the proper tissue remodeling. Reversion-inducing cysteine-rich protein with Kazal motifs (RECK) is a key regulator of ECM homeostasis and turnover. However, its role in wound contraction is presently unknown. Here we describe that Transforming growth factor type β1 (TGF-β1), one of the main pro-fibrotic wound-healing promoting factors, decreases RECK expression in fibroblasts through the Smad and JNK dependent pathways. This TGF-β1 dependent downregulation of RECK occurs with the concomitant increase of β1-integrin, which is required for fibroblasts adhesion and wound contraction through the activation of focal adhesion kinase (FAK). Loss and gain RECK expression experiments performed in different types of fibroblasts indicate that RECK downregulation mediates TGF-β1 dependent β1-integrin expression. Also, reduced levels of RECK potentiate TGF-β1 effects over fibroblasts FAK-dependent contraction, without affecting its cognate signaling. The above results were confirmed on fibroblasts derived from the Reck ^+/- mice compared to wild type-derived fibroblasts. We observed that Reck ^+/- mice heal dermal wounds more efficiently than wild type mice. Our results reveal a critical role for RECK in skin wound contraction as a key mediator in the axis: TGF-β1—RECK- β1-integrin.     

Microfossils,a Key to Unravel Cold-Water Carbonate Mound Evolution through Time: Evidence from the Eastern Alboran Sea

Cold-water coral (CWC) ecosystems occur worldwide and play a major role in the ocean''s carbonate budget and atmospheric CO₂ balance since the Danian (~65 m.y. ago). However their temporal and spatial evolution against climatic and oceanographic variability is still unclear. For the first time, we combine the main macrofaunal components of a sediment core from a CWC mound of the Melilla Mounds Field in the Eastern Alboran Sea with the associated microfauna and we highlight the importance of foraminifera and ostracods as indicators of CWC mound evolution in the paleorecord. Abundances of macrofauna along the core reveal alternating periods dominated by distinct CWC taxa (mostly Lophelia pertusa, Madrepora oculata) that correspond to major shifts in foraminiferal and ostracod assemblages. The period dominated by M. oculata coincides with a period characterized by increased export of refractory organic matter to the seafloor and rather unstable oceanographic conditions at the benthic boundary layer with periodically decreased water energy and oxygenation, variable bottom water temperature/density and increased sediment flow. The microfaunal and geochemical data strongly suggest that M. oculata and in particular Dendrophylliidae show a higher tolerance to environmental changes than L. pertusa. Finally, we show evidence for sustained CWC growth during the Alleröd-Younger-Dryas in the Eastern Alboran Sea and that this period corresponds to stable benthic conditions with cold/dense and well oxygenated bottom waters, high fluxes of labile organic matter and relatively strong bottom currents     

Chromosomal Evolution and Patterns of Introgression in Helianthus

Knowledge of the nature and extent of karyotypic differences between species provides insight into the evolutionary history of the genomes in question and, in the case of closely related species, the potential for genetic exchange between taxa. We constructed high-density genetic maps of the silverleaf sunflower (Helianthus argophyllus) and Algodones Dune sunflower (H. niveus ssp. tephrodes) genomes and compared them to a consensus map of cultivated sunflower (H. annuus) to identify chromosomal rearrangements between species. The genetic maps of H. argophyllus and H. niveus ssp. tephrodes included 17 linkage groups each and spanned 1337 and 1478 cM, respectively. Comparative analyses revealed greater divergence between H. annuus and H. niveus ssp. tephrodes (13 inverted segments, 18 translocated segments) than between H. annuus and H. argophyllus (10 inverted segments, 8 translocated segments), consistent with their known phylogenetic relationships. Marker order was conserved across much of the genome, with 83 and 64% of the H. argophyllus and H. niveus ssp. tephrodes genomes, respectively, being syntenic with H. annuus. Population genomic analyses between H. annuus and H. argophyllus, which are sympatric across a portion of the natural range of H. annuus, revealed significantly elevated genetic structure in rearranged portions of the genome, indicating that such rearrangements are associated with restricted gene flow between these two species.     

Geographic distributions,host plants and biology of species in the genus Huequenia (Coleoptera: Cerambycidae), with new records from Southwestern Argentina

Araucaria trees as host plants of the longhorned beetle Huequenia livida (Coleoptera: Cerambycidae) in Argentina are reviewed. Araucaria araucana is its natural host plant in SW Argentina, but the larvae also developed in dead branches of A. angustifolia and A. bidwillii (new host plant records), when both plants were kept in the same rearing cage with the natural host plant. Pinus contorta var. murrayana, also mentioned from Argentina, may be a recently adopted secondary host. A winter and a summer generation of H. livida was documented for the first time. Huequenia livida exceeds the actual natural distribution of A. araucana following the distribution of cultivated A. araucana and Pinus trees.