Cell-to-cell spread of Borna disease virus proceeds in the absence of the virus primary receptor and furin-mediated processing of the virus surface glycoprotein

Borna disease virus (BDV) is an enveloped virus with a nonsegmented negative-strand RNA genome whose organization is characteristic of Mononegavirales. BDV cell entry follows a receptor-mediated endocytosis pathway, which is initiated by the recognition of an as-yet-unidentified receptor at the cell surface by the virus glycoprotein G. BDV G is synthesized as a precursor (GPC) that is cleaved by the cellular protease furin to produce the mature glycoproteins GP1 and GP2, which have been implicated in receptor recognition and pH-dependent fusion events, respectively. BDV is highly neurotropic and its spread in cultured cells proceeds in the absence of detectable extracellular virus or syncytium formation. BDV spread has been proposed to be strictly dependent on the expression and correct processing of BDV G. Here we present evidence that cell-to-cell spread of BDV required neither the expression of cellular receptors involved in virus primary infection, nor the furin-mediated processing of BDV G. We also show that in furin-deficient cells, the release of BDV particles induced by the treatment of BDV-infected cells with hypertonic buffer was not significantly affected, while virion infectivity was dramatically impaired, correlating with the decreased incorporation of BDV G species into viral particles. These findings support the view that the propagation of BDV within the central nervous systems of infected hosts involves both a primary infection that follows a receptor-mediated endocytosis pathway and a subsequent cell-to-cell spread that is independent of the expression of the primary receptor and does not require the processing of BDV G into GP1 and GP2.     

Vasoactive intestinal peptide enhances growth and angiogenesis of human experimental prostate cancer in a xenograft model

We show that vasoactive intestinal peptide (VIP) exerts trophic and proangiogenic activities in experimental prostate cancer in vivo. Nude mice were subcutaneously injected with Matrigel impregnated with LNCaP prostate cancer cells. Cell treatment with 100 nM VIP for 1h before xenograft resulted in increased tumor growth after 8 and, more remarkably, 15 days of injection. The same occurred with the mRNA expression of the main angiogenic factor, vascular endothelial growth factor (VEGF), as shown by real-time RT-PCR quantification. The proangiogenic activity of VIP was further established by showing increases of hemoglobin levels, Masson trichromic staining, and immunohistochemical CD34 staining in tumors excised 15 days after subcutaneous injection of VIP-treated cells as compared to control conditions. All these parameters indicate that VIP increases vessel formation. This xenograft model is a useful tool to study in vivo the effects of VIP-related peptides in tumor growth and development of blood supply as well as their therapeutical potential in prostate cancer.     

Phylogenetic reconstruction from non-genomic data

MOTIVATION: Recent results related to horizontal gene transfer suggest that phylogenetic reconstruction cannot be determined conclusively from sequence data, resulting in a shift from approaches based on polymorphism information in DNA or protein sequence to studies aimed at understanding the evolution of complete biological processes. The increasing amount of available information on metabolic pathways for several species makes it of greater relevance to understand the similarities and differences among such pathways. These similarities can then be used to infer phylogenetic trees not based exclusively in sequence data, therefore avoiding the previously mentioned problems. RESULTS: In this article, we present a method to assess the structural similarity of metabolic pathways for several organisms. Our algorithms work by using one of the three possible enzyme similarity measures (hierarchical, information content, gene ontology), and one of the two clustering methods (neighbor-joining, unweighted pair group method with arithmetic mean), to produce a phylogenetic tree both in Newick and graphic format. The web server implementing our algorithms is optimized to answer queries in linear time. AVAILABILITY: The software is available for free public use on a web server, at the address http://www.jaist.ac.jp/~clemente/cgi-bin/phylo.pl. It is available on demand in source code form for research use to educational institutions, non-profit research institutes, government research laboratories and individuals, for non-exclusive use, without the right of the licensee to further redistribute the source code.     

Crystal structure of a soluble form of human monoglyceride lipase in complex with an inhibitor at 1.35 Å resolution

A high-resolution structure of a ligand-bound, soluble form of human monoglyceride lipase (MGL) is presented. The structure highlights a novel conformation of the regulatory lid-domain present in the lipase family as well as the binding mode of a pharmaceutically relevant reversible inhibitor. Analysis of the structure lacking the inhibitor indicates that the closed conformation can accommodate the native substrate 2-arachidonoyl glycerol. A model is proposed in which MGL undergoes conformational and electrostatic changes during the catalytic cycle ultimately resulting in its dissociation from the membrane upon completion of the cycle. In addition, the study outlines a successful approach to transform membrane associated proteins, which tend to aggregate upon purification, into a monomeric and soluble form.     

Light and ultrastructural description of Meglitschia mylei n. sp. (myxozoa) from Myleus rubripinnis (Teleostei: Serrasalmidae) in the Amazon River system

Meglitschia mylei n. sp. found in the gall bladder of the teleostean fish Myleus rubripinnis (Serrasalmidae) from the middle Amazonian region of Brazil is described using light and transmission electron microscopy. The spores observed in the bile averaged 24.6±0.8 μm long, 8.7±0.4 μm wide and 5.1±0.3 μm thick and were strongly furcate and arcuate ∩-shaped composed of two symmetric equal-sized valves, up to ～70 nm thick. Each valve possessed one opposed tapering appendage, 20.1±0.7 μm long, oriented parallel towards the basal tip of the appendages and joined along a right suture line forming a thick strand. The strand goes around the central part of the spore, which in turn surrounds two equal and symmetric spherical polar capsules (PC), 2.1±0.3 μm in diameter, located at the same level. Each capsule contains a polar filament with five (rarely six) coils. The binucleate sporoplasm was irregular in shape, contained several sporoplasmosomes, ～175 nm in diameter and filled all the space of the two caudal appendages. Based on the arc shape of the spore with two tapering caudal appendages oriented to the basis of spores, on the number and position of the PC and of the polar filament coils and arrangements, and on the host specificity, we propose the name M. mylei n. sp. for this new myxozoan. Accordingly, this is the second described species of this genus.     

Evaluating models to assess the distribution of Buxus balearica in southern Spain

Question: Which is the best model to predict the habitat distribution of Buxus balearica Lam. in southern Spain? Location: Málaga and Granada, Spain, across an area of 38 180 km². Methods: Prediction models based on 17 environmental variables were tested. Six methods were compared: multivariate adaptive regression spline (MARS), maximum entropy approach to modelling species' distributions (Maxent), two generic algorithms based on environmental metrics dissimilarity (BIOCLIM and DOMAIN), Genetic Algorithm for Rule‐set Prediction (GARP), and supervised learning methods based on generalized linear classifiers (support vector machines, SVMs). To test the predictive power of the models we used the Kappa index. Results: Maxent most accurately predicted the habitat distribution of B. balearica, followed by MARS models. The other models tested yielded lower accuracy values. A comparison of the predictive power of the models revealed that climate variables made the highest contributions among the environmental variables studied. The variables that made the lowest contributions were the insolation models. To examine the sensitivity of the models to a reduction in the number of variables, a test showed that accuracy of over 0.90 was maintained by applying just three climatic variables (spring rainfall, mean temperature of the warmest month, and mean temperature of the coldest month). Maps derived from the algorithms of all models tested coincided well with the known distribution of the species. Conclusions: Model habitat prediction is a preliminary step towards highlighting areas of high habitat suitability of B. balearica. These data support the results of previous research, which show that MaxEnt is the best technique for modelling species distributions with small sample sizes.     

Warm-up optimization in amateur male soccer players: A comparison of small-sided games and traditional warm-up routines on physical fitness qualities

The aim of this study was to compare the effects of small-sided soccer games (SSSGs) and traditional warm-up (TWU) routines on physical fitness qualities in soccer players. Following a between-subject, randomized design, amateur-level soccer players were assigned to a SSSG warm-up (n = 10; age: 19.3 ± 2.8 years) or TWU group (n = 10; age: 19.3 ± 2.4 years). Players completed multiple trials of 10-m and 30-m linear sprints, change-of-direction speed (CODS) tests, and countermovement jumps (CMJ) prior to and following the warm-up routine. Separate mixed ANOVAs were performed to assess group effects (SSSG vs. TWU), time effects within each group (pre- vs. post-warm-up), and their interaction for each physical fitness quality. No significant interaction effects were observed for any dependent variable. Significant improvements were evident between baseline and follow-up measurements for 10-m sprint time (p = 0.002, Hedges’ g effect size [g] = 0.59) and CMJ variables (height: p = 0.016, g = 0.20; power: p = 0.003, g = 0.19; force: p = 0.002, g = 0.14) in the TWU group and for CODS performance time (p = 0.012, g = 0.51) and CMJ variables (height: p < 0.001, g = 0.46; power: p = 0.002, g = 0.35; force: p = 0.001, g = 0.27) in the SSSG warm-up group. Both SSSG and TWU protocols improved selected physical fitness qualities with SSSG more effective at improving CODS and CMJ performance, and TWU more effective at improving linear speed. Soccer coaches may choose between SSSG or traditional warm-up activities according to player needs and preferences; however, the superior effects of SSSG suggest it might offer greater benefits than TWU in preparing players for optimal physical output.     

Multiple forms of lactate dehydrogenase in Staphylococcus aureus

Activities for nicotinamide adenine dinucleotide (NAD)-dependent and NAD-independent forms of lactate dehydrogenase (LDH) were measured in cell-free extracts of Staphylococcus aureus strain PS 6 for the d and l isomers of lactate. Data obtained for the NAD-dependent lactate dehydrogenases indicate that oxidation of both isomers of lactate is due to both an l-lactate-specific LDH and a lactate racemase. After acrylamide gel electrophoresis, two bands exhibiting LDH activity were detected in crude or in partially purified cell-free extracts. The fast band exhibited LDH activity that was not NAD-dependent for both isomers of lactate, whereas, the slow band had very high NAD-dependent LDH activity for the l isomer but just detectable activity or the d isomer. Both bands appeared when d-lactate was used as the substrate, but only the slow band was formed when l-lactate was the substrate. NAD-dependent LDH, in apparent association with a nonspecific tetrazolium-reducing protein, is responsible for the production of the slow band.