Road-associated edge effects in Amazonia change termite community composition by modifying environmental conditions

Roads and road-building are among the most important environmental impacts on forests near urban areas, but their effects on ecosystem processes and species distributions remain poorly known. Termites are the primary decomposer organisms in tropical forests and their spatial distribution is strongly affected by vegetation and soil structure. We studied the impacts of road construction on termite community structure in an Amazonian forest fragment near Manaus, Brazil. One leading question was whether the fragment under study was large enough to maintain the termite species pool present in nearby continuous forests. We also asked how soil moisture and canopy openness varied with proximity to roads, and whether these changes were associated with changes in termite species richness and composition in the fragment. While the forest fragment had a termite composition very similar to that of continuous forests, roads caused important changes in soil moisture and canopy openness, especially when close to forest edges. At distances of up to 81 m from roads, changes in soil moisture were significantly related to changes in termite species composition, but there was no correlation between canopy openness and species richness or composition. These results suggest that fragmentation caused by roads impacts termites in a different and less damaging manner than fragmentation caused by other kinds of degradation, and that even fragments bisected by roads can support very diverse communities and even undescribed taxa of termites. We conclude that a buffer zone should be established for conservation purposes in the reserves surrounded by roads.     

Molecular cross-talk between sponge host and associated microbes

Marine organisms especially those that live sessile, as sponges, are well known to have specific relationships with a great variety of microorganisms including bacteria and fungi. As most simple metazoan phylum, the Porifera, which emerged first during the transition from the non-Metazoa to the Metazoa from the common ancestor, comprise wide arrays of recognition molecules, both for Gram-negative bacteria and for Gram-positive bacteria as well as for fungi. They react specifically with effector molecules to inhibit or kill the invading microorganisms. The elicitation and the subsequent effector reactions of the sponges towards these microbes are outlined. However, besides of the elimination of bacteria and fungi, some of those taxa are kept as symbionts of the sponges, allowing them, for example, to accumulate the essential element manganese or to synthesize carotinoids. The sponges produce low-molecular-weight bioactive compounds, secondary metabolites, to eliminate the microorganisms. In addition, they are armed with cationic antimicrobial peptides allowing them to defend against invasive microorganisms and, in parallel, to kill or repel also metazoan invaders. The broad range of chemically and functionally different compounds qualifies the Porifera as the most important animal phylum to be exploited as a source for the isolation of new potential drugs. First molecular biological strategies have been outlined to obtain those compounds in a sustainable way, by producing them recombinantly.     

LPS Unmasking of Shigella flexneri Reveals Preferential Localisation of Tagged Outer Membrane Protease IcsP to Septa and New Poles

The Shigella flexneri outer membrane (OM) protease IcsP (SopA) is a member of the enterobacterial Omptin family of proteases which cleaves the polarly localised OM protein IcsA that is essential for Shigella virulence. Unlike IcsA however, the specific localisation of IcsP on the cell surface is unknown. To determine the distribution of IcsP, a haemagglutinin (HA) epitope was inserted into the non-essential IcsP OM loop 5 using Splicing by Overlap Extension (SOE) PCR, and IcsP^HA was characterised. Quantum Dot (QD) immunofluorescence (IF) surface labelling of IcsP^HA was then undertaken. Quantitative fluorescence analysis of S. flexneri 2a 2457T treated with and without tunicaymcin to deplete lipopolysaccharide (LPS) O antigen (Oag) showed that IcsP^HA was asymmetrically distributed on the surface of septating and non-septating cells, and that this distribution was masked by LPS Oag in untreated cells. Double QD IF labelling of IcsP^HA and IcsA showed that IcsP^HA preferentially localised to the new pole of non-septating cells and to the septum of septating cells. The localisation of IcsP^HA in a rough LPS S. flexneri 2457T strain (with no Oag) was also investigated and a similar distribution of IcsP^HA was observed. Complementation of the rough LPS strain with rmlD resulted in restored LPS Oag chain expression and loss of IcsP^HA detection, providing further support for LPS Oag masking of surface proteins. Our data presents for the first time the distribution for the Omptin OM protease IcsP, relative to IcsA, and the effect of LPS Oag masking on its detection.     

DNA sequence analysis to guide the release of blue-and-yellow macaws (Ara ararauna, Psittaciformes, Aves) from the illegal trade back into the wild

The illegal wildlife trade is one of the major threats to Brazil’s biodiversity. Approximately 80 % of illegally captured animals are birds, and 4 % of those are parrots. Although many seized birds do not survive, those that are recovered may be returned to the wild. The release of seized individuals into the wild should be conducted with caution, as local populations may suffer adverse effects if genetically different individuals are introduced. In this study, we evaluated the genetic relationships between 13 illegally captured blue-and-yellow macaws selected for release in northeastern Goiás, Brazil, and previously studied Brazilian macaw populations. We identified the seized macaws that were genetically similar to those from northwestern Goiás and that were therefore most suitable for release in that area. The genetic relationship was evaluated by sequence analysis of 403 bp of mitochondrial DNA control region. Relationships between mitochondrial haplotypes were computed via a median-joining network. Only six of the seized macaws were closely related to the macaws of northeastern Goiás, indicating that those macaws were potential candidates for release in that area. However, the release of these birds should follow all technical recommendations required by the Brazilian environmental authorities.     

Mouriri morleyii sp. nov. (Melastomataceae) from Brazil,with notes on its foliar stomatal crypts

Mouriri morleyii R. Goldenb. & Meirelles sp. nov. can be distinguished from the other species in the genus by its large stomatal crypts (the largest ones in the genus), columnar sclereids, tetramerous flowers and calyx closed in bud. This new species was collected on an inselberg in the state of Espírito Santo, Brazil.     

In Silico Modeling of Shear-Stress-Induced Nitric Oxide Production in Endothelial Cells through Systems Biology

Nitric oxide (NO) produced by vascular endothelial cells is a potent vasodilator and an antiinflammatory mediator. Regulating production of endothelial-derived NO is a complex undertaking, involving multiple signaling and genetic pathways that are activated by diverse humoral and biomechanical stimuli. To gain a thorough understanding of the rich diversity of responses observed experimentally, it is necessary to account for an ensemble of these pathways acting simultaneously. In this article, we have assembled four quantitative molecular pathways previously proposed for shear-stress-induced NO production. In these pathways, endothelial NO synthase is activated 1), via calcium release, 2), via phosphorylation reactions, and 3), via enhanced protein expression. To these activation pathways, we have added a fourth, a pathway describing actual NO production from endothelial NO synthase and its various protein partners. These pathways were combined and simulated using CytoSolve, a computational environment for combining independent pathway calculations. The integrated model is able to describe the experimentally observed change in NO production with time after the application of fluid shear stress. This model can also be used to predict the specific effects on the system after interventional pharmacological or genetic changes. Importantly, this model reflects the up-to-date understanding of the NO system, providing a platform upon which information can be aggregated in an additive way.     

Dual inhibition of DNA polymerase PolC and protein tyrosine phosphatase CpsB uncovers a novel antibiotic target

Increasing antibiotic resistance is making the identification of novel antimicrobial targets critical. Recently, we discovered an inhibitor of protein tyrosine phosphatase CpsB, fascioquinol E (FQE), which unexpectedly inhibited the growth of Gram-positive pathogens. CpsB is a member of the polymerase and histidinol phosphate phosphatase (PHP) domain family. Another member of this family found in a variety of Gram-positive pathogens is DNA polymerase PolC. We purified the PHP domain from PolC (PolC_PHP), and showed that this competes away FQE inhibition of CpsB phosphatase activity. Furthermore, we showed that this domain hydrolyses the 5′-p-nitrophenyl ester of thymidine-5′-monophosphate (pNP-TMP), which has been used as a measure of exonuclease activity. Finally, we showed that FQE not only inhibits the phosphatase activity of CpsB, but also ability of PolC_PHP to catalyse the hydrolysis of pNP-TMP. This suggests that PolC may be the essential target of FQE, and that the PHP domain may represent an as yet untapped target for the development of novel antibiotics.     

Escherichia coli DnaE Polymerase Couples Pyrophosphatase Activity to DNA Replication

DNA Polymerases generate pyrophosphate every time they catalyze a step of DNA elongation. This elongation reaction is generally believed as thermodynamically favoured by the hydrolysis of pyrophosphate, catalyzed by inorganic pyrophosphatases. However, the specific action of inorganic pyrophosphatases coupled to DNA replication in vivo was never demonstrated. Here we show that the Polymerase-Histidinol-Phosphatase (PHP) domain of Escherichia coli DNA Polymerase III α subunit features pyrophosphatase activity. We also show that this activity is inhibited by fluoride, as commonly observed for inorganic pyrophosphatases, and we identified 3 amino acids of the PHP active site. Remarkably, E. coli cells expressing variants of these catalytic residues of α subunit feature aberrant phenotypes, poor viability, and are subject to high mutation frequencies. Our findings indicate that DNA Polymerases can couple DNA elongation and pyrophosphate hydrolysis, providing a mechanism for the control of DNA extension rate, and suggest a promising target for novel antibiotics.