Apoptotic neutrophils release macrophage migration inhibitory factor upon stimulation with tumor necrosis factor-alpha

Macrophage migration inhibitory factor (MIF) is an important cytokine involved in the regulation of innate immunity and present at increased levels during inflammatory responses. Here we demonstrate that mature blood and tissue neutrophils constitutively express MIF as a cytosolic protein not associated with azurophil granules. Functionally active MIF, but not proteases stored in azurophil granules, was released from apoptotic neutrophils following short term tumor necrosis factor (TNF)-alpha stimulation in a caspase-dependent manner and prior to any detectable phagocytosis by monocyte-derived macrophages. Moreover, TNF-alpha-mediated MIF release was blocked by glyburide and propenicide, both inhibitors of ATP-binding cassette-type transporters, suggesting that this transporter system is activated during neutrophil apoptosis. Taken together, apoptotic mature neutrophils release MIF upon short term TNF-alpha stimulation. Therefore, apoptosis may not always occur without the induction of pro-inflammatory mechanisms.     

Blood volume and haemoglobin oxygen content changes in human bone marrow during orthostatic stress

The interest in, and the need for effective measures to be used in screening, diagnosis, and the follow-up of skeletal pathologies is growing markedly. This paper proposes a completely new and non-invasive technique allowing the study of the human tibia bone marrow (BM) haemodynamics with a time resolution of 1 s. The technique, based on near infrared spectroscopy, is sensitive enough to allow the detection of BM blood volume and/or oxygen saturation changes during orthostatic variations imposed by a tilt bed. An increase in the slope of the bed of 15 degrees is sufficient to detect this phenomenon. The ability to study the possible presence of a neural control of BM haemodynamics is also discussed. No other existing technique currently allows one to obtain the proposed results and this approach might open up a new field of study related to human BM physiology.     

Platelet-activating factor receptor plays a role in lung injury and death caused by Influenza A in mice

Influenza A virus causes annual epidemics which affect millions of people worldwide. A recent Influenza pandemic brought new awareness over the health impact of the disease. It is thought that a severe inflammatory response against the virus contributes to disease severity and death. Therefore, modulating the effects of inflammatory mediators may represent a new therapy against Influenza infection. Platelet activating factor (PAF) receptor (PAFR) deficient mice were used to evaluate the role of the gene in a model of experimental infection with Influenza A/WSN/33 H1N1 or a reassortant Influenza A H3N1 subtype. The following parameters were evaluated: lethality, cell recruitment to the airways, lung pathology, viral titers and cytokine levels in lungs. The PAFR antagonist PCA4248 was also used after the onset of flu symptoms. Absence or antagonism of PAFR caused significant protection against flu-associated lethality and lung injury. Protection was correlated with decreased neutrophil recruitment, lung edema, vascular permeability and injury. There was no increase of viral load and greater recruitment of NK1.1(+) cells. Antibody responses were similar in WT and PAFR-deficient mice and animals were protected from re-infection. Influenza infection induces the enzyme that synthesizes PAF, lyso-PAF acetyltransferase, an effect linked to activation of TLR7/8. Therefore, it is suggested that PAFR is a disease-associated gene and plays an important role in driving neutrophil influx and lung damage after infection of mice with two subtypes of Influenza A. Further studies should investigate whether targeting PAFR may be useful to reduce lung pathology associated with Influenza A virus infection in humans.     

Synechococcus: 3 billion years of global dominance

Cyanobacteria are among the most important primary producers on the Earth. However, the evolutionary forces driving cyanobacterial species diversity remain largely enigmatic due to both their distinction from macro‐organisms and an undersampling of sequenced genomes. Thus, we present a new genome of a Synechococcus‐like cyanobacterium from a novel evolutionary lineage. Further, we analyse all existing 16S rRNA sequences and genomes of Synechococcus‐like cyanobacteria. Chronograms showed extremely polyphyletic relationships in Synechococcus, which has not been observed in any other cyanobacteria. Moreover, most Synechococcus lineages bifurcated after the Great Oxidation Event, including the most abundant marine picoplankton lineage. Quantification of horizontal gene transfer among 70 cyanobacterial genomes revealed significant differences among studied genomes. Horizontal gene transfer levels were not correlated with ecology, genome size or phenotype, but were correlated with the age of divergence. All findings were synthetized into a novel model of cyanobacterial evolution, characterized by serial convergence of the features, that is multicellularity and ecology.     

Impact of the isoelectric point of model parvoviruses on viral retention in anion‐exchange chromatography

Anion‐exchange chromatography (AEX) is used in the downstream purification of monoclonal antibodies to remove impurities and potential viral contamination based on electrostatic interactions. Although the isoelectric point (pI) of viruses is considered a key factor predicting the virus adsorption to the resin, the precise molecular mechanisms involved remain unclear. To address this question, we compared structurally homologous parvoviruses that only differ in their surface charge distribution. A single charged amino acid substitution on the capsid surface of minute virus of mice (MVM) provoked an increased apparent pI (pI_app) 6.2 compared to wild‐type MVM (pI_app = 4.5), as determined by chromatofocusing. Despite their radically different pI_app, both viruses displayed the same interaction profile in Mono Q AEX at different pH conditions. In contrast, the closely related canine parvovirus (pI_app = 5.3) displayed a significantly different interaction at pH 5. The detailed structural analysis of the intricate three‐dimensional structure of the capsids suggests that the charge distribution is critical, and more relevant than the pI, in controlling the interaction of a virus with the chromatographic resin. This study contributes to a better understanding of the molecular mechanisms governing virus clearance by AEX, which is crucial to enable robust process design and maximize safety.     

Invasive spread in meta-food-webs depends on landscape structure,fertilization and species characteristics

Land use change and biological invasions collectively threaten biodiversity. Yet, few studies have addressed how altering the landscape structure and nutrient supply can promote biological invasions and particularly invasive spread (the spread of an invader from the place of introduction), or asked whether and how these factors interact with biotic interactions and invader properties. We here bridge this knowledge gap by providing a holistic network-based approach. Our approach combines a trophic network model with a spatial network model allowing us to test which combinations of abiotic and biotic factors can facilitate invasions and in particular invasive spread in food webs. We numerically simulated 6300 single-species invasions in clustered and random landscapes at different levels of nutrient supply. In total, our simulation experiment yielded 69% successful invasions – 71% in clustered landscapes and 66% in random landscapes, with the proportion of successful invasions increasing with nutrient supply. However, invasive spread was generally higher in random than in clustered landscapes. The latter can facilitate invasive spread within a habitat cluster, but prevent invasive spread between clusters. Low nutrient levels generally prevented the establishment of invasive species and their subsequent spread. However, successful invaders could have more severe impacts as they contribute more to total biomass density and species richness under such conditions. Good dispersal abilities drive the broad-scale spread of invasive species in fragmented landscapes. Our approach makes an important contribution towards a better understanding of what combination of landscape and invader properties can facilitate or prevent invasive spread in natural ecosystems. This should allow ecologists to more effectively predict and manage biological invasions.     

The Interaction of Large Amplitude Internal Seiches with a Shallow Sloping Lakebed: Observations of Benthic Turbulence in Lake Simcoe,Ontario, Canada

Observations of the interactions of large amplitude internal seiches with the sloping boundary of Lake Simcoe, Canada show a pronounced asymmetry between up- and downwelling. Data were obtained during a 42-day period in late summer with an ADCP and an array of four thermistor chains located in a 5 km line at the depths where the thermocline intersects the shallow slope of the lakebed. The thermocline is located at depths of 12–14 m during the strongly stratified period of late summer. During periods of strong westerly winds the thermocline is deflected as much as 8 m vertically and interacts directly with the lakebed at depth between 14–18 m. When the thermocline was rising at the boundary, the stratification resembles a turbulent bore that propagates up the sloping lakebed with a speed of 0.05–0.15 m s⁻¹ and a Froude number close to unity. There were strong temperature overturns associated with the abrupt changes in temperature across the bore. Based on the size of overturns in the near bed stratification, we show that the inferred turbulent diffusivity varies by up to two orders of magnitude between up- and downwellings. When the thermocline was rising, estimates of turbulent diffusivity were high with K_Z ∼10⁻⁴ m²s⁻¹, whereas during downwelling events the near-bed stratification was greatly increased and the turbulence was reduced. This asymmetry is consistent with previous field observations and underlines the importance of shear-induced convection in benthic bottom boundary layers of stratified lakes.     

Structure of p53 binding to the BAX response element reveals DNA unwinding and compression to accommodate base-pair insertion

The p53 core domain binds to response elements (REs) that contain two continuous half-sites as a cooperative tetramer, but how p53 recognizes discontinuous REs is not well understood. Here we describe the crystal structure of the p53 core domain bound to a naturally occurring RE located at the promoter of the Bcl-2-associated X protein (BAX) gene, which contains a one base-pair insertion between the two half-sites. Surprisingly, p53 forms a tetramer on the BAX-RE that is nearly identical to what has been reported on other REs with a 0-bp spacer. Each p53 dimer of the tetramer binds in register to a half-site and maintains the same protein–DNA interactions as previously observed, and the two dimers retain all the protein–protein contacts without undergoing rotation or translation. To accommodate the additional base pair, the DNA is deformed and partially disordered around the spacer region, resulting in an apparent unwinding and compression, such that the interactions between the dimers are maintained. Furthermore, DNA deformation within the p53-bound BAX-RE is confirmed in solution by site-directed spin labeling measurements. Our results provide a structural insight into the mechanism by which p53 binds to discontinuous sites with one base-pair spacer.