The Atomic Structure of the HIV-1 gp41 Transmembrane Domain and Its Connection to the Immunogenic Membrane-proximal External Region

The membrane-proximal external region (MPER) C-terminal segment and the transmembrane domain (TMD) of gp41 are involved in HIV-1 envelope glycoprotein-mediated fusion and modulation of immune responses during viral infection. However, the atomic structure of this functional region remains unsolved. Here, based on the high resolution NMR data obtained for peptides spanning the C-terminal segment of MPER and the TMD, we report two main findings: (i) the conformational variability of the TMD helix at a membrane-buried position; and (ii) the existence of an uninterrupted α-helix spanning MPER and the N-terminal region of the TMD. Thus, our structural data provide evidence for the bipartite organization of TMD predicted by previous molecular dynamics simulations and functional studies, but they do not support the breaking of the helix at Lys-683, as was suggested by some models to mark the initiation of the TMD anchor. Antibody binding energetics examined with isothermal titration calorimetry and humoral responses elicited in rabbits by peptide-based vaccines further support the relevance of a continuous MPER-TMD helix for immune recognition. We conclude that the transmembrane anchor of HIV-1 envelope is composed of two distinct subdomains: 1) an immunogenic helix at the N terminus also involved in promoting membrane fusion; and 2) an immunosuppressive helix at the C terminus, which might also contribute to the late stages of the fusion process. The unprecedented high resolution structural data reported here may guide future vaccine and inhibitor developments.     

No Major Host Genetic Risk Factor Contributed to A(H1N1)2009 Influenza Severity

While most patients affected by the influenza A(H1N1) pandemic experienced mild symptoms, a small fraction required hospitalization, often without concomitant factors that could explain such a severe course. We hypothesize that host genetic factors could contribute to aggravate the disease. To test this hypothesis, we compared the allele frequencies of 547,296 genome-wide single nucleotide polymorphisms (SNPs) between 49 severe and 107 mild confirmed influenza A cases, as well as against a general population sample of 549 individuals. When comparing severe vs. mild influenza A cases, only one SNP was close to the conventional p = 5×10⁻⁸. This SNP, rs28454025, sits in an intron of the GSK233 gene, which is involved in a neural development, but seems not to have any connections with immunological or inflammatory functions. Indirectly, a previous association reported with CD55 was replicated. Although sample sizes are low, we show that the statistical power in our design was sufficient to detect highly-penetrant, quasi-Mendelian genetic factors. Hence, and assuming that rs28454025 is likely to be a false positive, no major genetic factor was detected that could explain poor influenza A course.     

Characterization of Prorocentrum elegans and Prorocentrum levis (Dinophyceae) from the southeastern Bay of Biscay by morphology and molecular phylogeny

Benthic Prorocentrum species can produce toxins that adversely affect animals and human health. They are known to co‐occur with other bloom‐forming, potentially toxic, benthic dinoflagellates of the genera Ostreopsis, Coolia, and Gambierdiscus. In this study, we report on the presence of P. elegans M.Faust and P. levis M.A.Faust, Kibler, Vandersea, P.A. Tester & Litaker from the southeastern Bay of Biscay. Sampling was carried out in the Summer‐Autumn 2010–2012 along the Atlantic coast of the Iberian Peninsula, but these two species were only found in the northeastern part of the Peninsula. Strains were isolated from macroalgae collected from rocky‐shore areas bordering accessible beaches. Morphological traits of isolated strains were analyzed by LM and SEM, whereas molecular analyses were performed using the LSU and internal transcribed spacer (ITS)1‐5.8S‐ITS2 regions of the rDNA. A bioassay with Artemia fransciscana and liquid chromatography–high‐resolution mass spectrometry analyses were used to check the toxicity of the species, whose results were negative. The strains mostly corresponded to their species original morphological characterization, which is supported by the phylogenetic analyses in the case of P. levis, whereas for P. elegans, this is the first known molecular characterization. This is also the second known report of P. elegans.     

Arthropod fauna on grassland–heathland associations under different grazing managements with domestic ruminants

The effects of two grazer species (cattle or sheep) and two flock types (single or mixed with goats) on vegetation and arthropod fauna were studied in a factorial design on eight plots which comprised two thirds of mechanically cleared heathland and one third of improved ryegrass-clover grassland. After six grazing seasons, the shrubland areas were dominated by gorse (Ulex gallii) in all treatments. Herbaceous cover was higher under mixed than under single grazing, and under sheep than under cattle grazing. Higher captures of Opiliones, Julida, Lithobiomorpha, Microcoryphia and Carabidae were recorded in shrublands than in grasslands, while the reverse was observed for Linyphiidae, Lycosidae and Hemiptera. Within shrublands, fauna responded to the flock type but not to the grazer species. More arthropod groups favoured the patchier areas with higher herbaceous biomass generated by mixed herds with goats. Within grasslands, species-specific responses to the grazer species were observed. Mixed grazing schemes which include goats within partially improved heathlands could contribute to maintain higher biodiversity levels in these marginal areas.     

Changes in heathland vegetation under goat grazing: effects of breed and stocking rate

Questions: How are heathland vegetation dynamics affected by different goat grazing management? Location: Cantabrian heathlands in Illano, Asturias, northern Spain. Methods: During 4 years, vegetation dynamics (structural composition, canopy height and floristic diversity) were studied under three goat grazing treatments with three replicates: high stocking rate (11.7 goats ha^?1) with a local Celtiberic breed, and high (15 goats ha^?1) and low (6.7 goats ha^?1) stocking rates with a commercial Cashmere breed. Results: The relative cover of woody plants, particularly heather species, decreased more while herbaceous cover increased more under local Celtiberic than under Cashmere breed grazing. Within Cashmere treatments, the cover and height of live shrubs decreased more and the herbaceous cover increased more under high than under low stocking rate. Redundancy analysis showed a significant effect of treatment × year interaction on floristic composition. Greater species richness was recorded under local goat grazing, but Shannon diversity index fell in the fourth year on these plots because of dominance by two grass species. Conclusions: Local Celtiberic goat grazing at such a high stocking rate (11.7 goats ha^?1) hinders the development of sustainable systems on these heathlands, both in environmental and productive terms, owing to the limitations in soil fertility. Nevertheless, Celtiberic goats could be useful for controlling excessive shrub encroachment and reducing fire hazard. Cashmere goat grazing at high stocking rate promoted the highest Shannon diversity by generating a better balance between woody and herbaceous plants, while shrub dominance was not altered under the low stocking rate.     

MHC class II DRB1 gene polymorphism in the pathogenesis of Maedi–Visna and pulmonary adenocarcinoma viral diseases in sheep

Ovine pulmonary adenocarcinoma (OPA) and Maedi–Visna (Maedi) are two chronic respiratory diseases of retroviral origin which occur worldwide. It is known that different host genetic factors influence the outcome of viral infections. To determine if variation in the Mhc-DRB1 gene was associated with progression to these ovine diseases, sheep lungs with and without OPA and Maedi lesions were collected. A sequence-based method was applied and 40 different alleles were detected in the sample analysed. In the allele-by-allele association analysis, allele DRB1*0325 had a significant association with susceptibility to Maedi (P = 0.045). For OPA, DRB1*0143 and DRB1*0323 were significantly associated with susceptibility (P = 0.024 and P = 0.029), and allele DRB1*0702 was significantly associated with resistance (P = 0.012). Based on these results, the Mhc-DRB1 alleles were classified by effect in three categories—susceptible (S), resistant (R) and neutral (N)—and animals were reassigned the genotypes as S/S, S/R, S/N, R/R, R/N and N/N. In a second analysis, penalised logistic regression models including a flock effect were run. In Maedi, significant association was detected for the N/S heterozygote (P = 0.0007), but not for the S/S homozygote, probably as a result of the low number of S/S animals. In OPA, association was detected for both the S/S and R/R homozygotes (P = 0.005 and P = 0.047). This allele grouping method may be applied in association studies with highly variable genes. This is the first study demonstrating significant associations between sheep Mhc-DRB1 alleles and susceptibility to OPA and Maedi. Therefore, both diseases are suitable candidates for more comprehensive genetic studies.