Viable neutrophils release mitochondrial DNA to form neutrophil extracellular traps

Neutrophil extracellular traps (NETs) represent extracellular structures able to bind and kill microorganisms. It is believed that they are generated by neutrophils undergoing cell death, allowing these dying or dead cells to kill microbes. We show that, following priming with granulocyte/macrophage colony-stimulating factor (GM-CSF) and subsequent short-term toll-like receptor 4 (TLR4) or complement factor 5a (C5a) receptor stimulation, viable neutrophils are able to generate NETs. Strikingly, NETs formed by living cells contain mitochondrial, but no nuclear, DNA. Pharmacological or genetic approaches to block reactive oxygen species (ROS) production suggested that NET formation is ROS dependent. Moreover, neutrophil populations stimulated with GM-CSF and C5a showed increased survival compared with resting neutrophils, which did not generate NETs. In conclusion, mitochondrial DNA release by neutrophils and NET formation do not require neutrophil death and do also not limit the lifespan of these cells.

Pollen percentage thresholds of Abies alba based on 13-year annual records of pollen deposition in modified Tauber traps: perspectives of application to fossil situations

Abies alba (fir), a submontane tree from Central European mountains and uplands, is of special interest for palaeoecological and palaeoclimate interpretations due to its sensitivity to air and soil humidity. Its present distribution limit in the uplands of SE Poland is still a matter of debate. In the Holocene fir expanded to Poland very late, but early fir populations are supposed to occur in the Šumava Mts (Czech Republic). The study aims: to estimate pollen thresholds for fir presence/absence in Bohemia (Czech Republic) and Poland on the basis of modified Tauber pollen traps; to use these thresholds for tracing fir presence in two pollen diagrams from Poland (Słone and Bezedna lakes) in the border zone between the Roztocze region (with fir forest stands today) and Polesie (where fir has never played an important role); and to investigate how the percentage presence/absence threshold can be used to trace the occurrence and abundance of fir trees in the Šumava Mts based on the pollen diagrams of Rokytecká slat' and Mrtvý luh. The fir pollen thresholds estimated in terms of PAR (pollen accumulation rates or pollen influx) range from 843 (grains cm− 2 year− 1) (Roztocze) to 61 (Krkonoše) and 49 (Šumava). Percentage thresholds range from 0.3% in Krkonoše where fir trees are not present within 4 km to 22% in fir-dominated woodland of the Roztocze, providing evidence of strong underrepresentation of fir in the pollen deposition. Application of these percentage thresholds to the Słone and Bezedna pollen diagrams indicates that occurrence of fir in the region is possible from 3.5 cal ky BP onwards, though the evidence is not decisive. In the Šumava, a low representation of fir pollen (1–2%) reflecting presence of scattered fir trees was detected as early as ca. 7.0 cal ky BP.

Extracellular DNA traps in allergic, infectzious, and autoimmune diseases

Extracellular DNA traps are part of the innate immune response and are seen with many infectious, allergic, and autoimmune diseases. They can be generated by several different leukocytes, including neutrophils, eosinophils, and monocytes, as well as mast cells. Here, we review the composition of these extracellular DNA-containing structures as well as potential mechanisms for their production and function. In general, extracellular DNA traps have been described as binding to and killing pathogens, particularly bacteria, fungi, but also parasites. On the other hand, it is possible that DNA traps contribute to immunopathology in chronic inflammatory diseases, such as bronchial asthma. In addition, it has been demonstrated that they can initiate and/or potentiate autoimmune diseases. Extracellular DNA traps represent a frequently observed phenomenon in inflammatory diseases, and they appear to participate in the cross-talk between different immune cells. These new insights into the pathogenesis of inflammatory diseases may open new avenues for targeted therapies.

NADPH oxidase-independent formation of extracellular DNA traps by basophils

Basophils are primarily associated with a proinflammatory and immunoregulatory role in allergic diseases and parasitic infections. Recent studies have shown that basophils can also bind various bacteria both in the presence and the absence of opsonizing Abs. In this report, we show that both human and mouse basophils are able to produce mitochondrial reactive oxygen species and to form extracellular DNA traps upon IL-3 priming and subsequent activation of the complement factor 5 a receptor or FcεRI. Such basophil extracellular traps (BETs) contain mitochondrial, but not nuclear DNA, as well as the granule proteins basogranulin and mouse mast cell protease 8. BET formation occurs despite the absence of any functional NADPH oxidase in basophils. BETs can be found in both human and mouse inflamed tissues, suggesting that they also play a role under in vivo inflammatory conditions. Taken together, these findings suggest that basophils exert direct innate immune effector functions in the extracellular space.

Impaired DNase1-mediated degradation of neutrophil extracellular traps is associated with acute thrombotic microangiopathies.

BACKGROUND Acute thrombotic microangiopathies (TMAs) are characterized by excessive microvascular thrombosis and are associated with markers of neutrophil extracellular traps (NETs) in plasma. NETs are composed of DNA fibers and promote thrombus formation through the activation of platelets and clotting factors. OBJECTIVE The efficient removal of NETs may be required to prevent excessive thrombosis such as in TMAs. To test this hypothesis, we investigated whether TMAs are associated with a defect in the degradation of NETs. APPROACH AND RESULTS We show that NETs generated in vitro were efficiently degraded by plasma from healthy donors. However, NETs remained stable after exposure to plasma from TMA patients. The inability to degrade NETs was linked to a reduced DNase activity in TMA plasma. Plasma DNase1 was required for efficient NET-degradation and TMA plasma showed decreased levels of this enzyme. Supplementation of TMA plasma with recombinant human DNase1 restored NET-degradation activity. CONCLUSIONS Our data indicates that DNase1-mediated degradation of NETs is impaired in patients with TMAs. The role of plasma DNases in thrombosis is, as of yet, poorly understood. Reduced plasma DNase1 activity may cause the persistence of pro-thrombotic NETs and thus promote microvascular thrombosis in TMA patients. This article is protected by copyright. All rights reserved.

Seven years of annual pollen influx at the forest limit in the Swiss Alps studied by pollen traps: relations to vegetation and climate

Annual pollen influx has been monitored in short transects across the altitudinal tree limit in four areas of the Swiss Alps with the use of modified Tauber traps placed at the ground surface. The study areas are Grindelwald (8 traps), Aletsch (8 traps), Simplon (5 traps), and Zermatt (5 traps). The vegetation around the traps is described. The results obtained are: (1) Peak years of pollen influx (one or two in seven years) follow years of high average air temperatures during June–November of the previous year for Larix and Picea, and less clearly for Pinus non-cembra, but not at all for Pinus cembra and Alnus viridis. (2) At the upper forest limit, the regional pollen influx of trees (trees absent within 100 m of the pollen trap) relates well to the average basal area of the same taxon within 10–15 km of the study areas for Pinus cembra, Larix, and Betula, but not for Picea, Pinus non-cembra, and Alnus viridis. (3) The example of Zermatt shows that pollen influx characterises the upper forest limit, if the latter is more or less intact. (4) Presence/absence of Picea, Pinus cembra, Larix, Pinus non-cembra, and Alnus viridis trees within 50–100 m of the traps is apparent in the pollen influx in peak years of pollen influx but not in other years, suggesting that forest-limit trees produce significant amounts of pollen only in some years. (5) Pollen influx averaged over the study period correlates well with the abundance of plants around the pollen traps for conifer trees (but not deciduous trees), Calluna, Gramineae, and Cyperaceae, and less clearly so Compositae Subfam. Cichorioideae and Potentilla-type. (6) Influx of extra-regional pollen derived from south of the Alps is highest in Simplon, which is open to southerly winds, slightly lower in Aletsch lying just north of Simplon, and lowest in Zermatt sheltered from the south by high mountains and Grindelwald lying north of the central Alps.

(Table 1) Chemical and isotopic analyses of Rajmahal Traps and Ninetyeast Ridge volcanics

The hypothesis that hotspots are the sources of many continental flood basalts is evaluated geochemically for the proposed Rajmahal Traps-Ninetyeast Ridge-Kerguelen hotspot system. It appears that the Kerguelen hotspot did not directly feed Rajmahal magmas, although it may have provided a source of heat for Rajmahal activity.

Ground beetle (Carabidae) abundance data from north-eastern Australian rainforest, between June 2008 - January 2010, using pitfall traps

Understanding how the environment influences patterns of diversity is vital for effective conservation management, especially in a changing global climate. While assemblage structure and species richness patterns are often correlated with current environmental factors, historical influences may also be considerable, especially for taxa with poor dispersal abilities. Mountain-top regions throughout tropical rainforests can act as important refugia for taxa characterised by low dispersal capacities such as flightless ground beetles (Carabidae), an ecologically significant predatory group. We surveyed flightless ground beetles along elevational gradients in five different subregions within the Australian Wet Tropics World Heritage Area to investigate (1) whether the diversity and composition of flightless ground beetles are elevationally stratified, and, if so, (2) what environmental factors (other than elevation per se) are associated with these patterns. Generalised linear models and model averaging techniques were used to relate patterns of diversity to environmental factors. Unlike most taxonomic groups, flightless ground beetles increased in species richness and abundance with elevation. Additionally, each subregion consisted of distinct assemblages containing a high level of regional endemic species. Species richness was most strongly positively associated with the historical climatic conditions and negatively associated with severity of recent disturbance (treefalls) and current climatic conditions. Assemblage composition was associated with latitude and current and historical climatic conditions. Our results suggest that distributional patterns of flightless ground beetles are not only likely to be associated with factors that change with elevation (current climatic conditions), but also factors that are independent of elevation (recent disturbance and historical climatic conditions). Variation in historical vegetation stability explained both species richness and assemblage composition patterns, probably reflecting the significance of upland refugia at a geographic time scale. These findings are important for conservation management as upland habitats are under threat from climate change.

Particle fluxes at sediment traps AU9701, south of Australia

Sediment trap moorings were deployed from September 21, 1997 through February 21, 1998 at three locations south of Australia along 140°E: at -47°S in the central Subantarctic Zone (SAZ) with traps at 1060, 2050, and 3850 m depth, at -51°S in the Subantarctic Front with one trap at 3080 m, and at -54°S in the Polar Frontal Zone (PFZ) with traps at 830 and 1580 m. Particle fluxes were high at all the sites (18-32 g/m**2/yr total mass and 0.5-1.4 g organic carbon/m**2/yr at -1000 m, assuming minimal flux outside the sampled summer period). These values are similar to other Southern Ocean results and to the median estimated for the global ocean by Lampitt and Antia [1997], and emphasize that the Southern Ocean exports considerable carbon to the deep sea despite its 'high-nutrient, low chlorophyll' characteristics. The SAZ site was dominated by carbonate (>50% of total mass) and the PFZ site by biogenic silica (>50% of total mass). Both sites exhibited high export in spring and late summer, with an intervening low flux period in December. For the 153 day collection period, particulate organic carbon export was somewhat higher in all the traps in the SAZ (range 0.57-0.84 gC/m**2) than in the PFZ (range 0.31-0.53), with an intermediate value observed at the SAF (0.60). The fraction of surface organic carbon export (estimated from seasonal nutrient depletion, Lourey and Trull [2001]) reaching 1000 m was indistinguishable in the SAZ and PFZ, despite different algal communities.

Fecal pellets flux at DYFAMED sediment traps

Because zooplankton feces represent a potentially important transport pathway of surface-derived organic carbon in the ocean, we must understand the patterns of fecal pellet abundance and carbon mobilization over a variety of spatial and temporal scales. To assess depth-specific water column variations of fecal pellets on a seasonal scale, vertical fluxes of zooplankton fecal pellets were quantified and their contribution to mass and particulate carbon were computed during 1990 at 200, 500, 1000, and 2000 m depths in the open northwestern Mediterranean Sea as part of the French-JGOFS DYFAMED Program. Depth-averaged daily fecal pellet flux was temporally variable, ranging from 3.04 * 10**4 pellets m**2/d in May to a low of 6.98 * 10**2 pellets m**2/d in September. The peak flux accounted for 50% of the integrated annual flux of fecal pellets and 62% of pellet carbon during only two months in mid-spring (April and May). Highest numerical fluxes were encountered at 1000 m, suggesting fecal pellet generation well below the euphotic zone. However, there was a trend toward lower pellet carbon with increasing depth, suggesting bacterial degradation or in situ repackaging as pellets sink through the water column. At 500 m, both the lowest pellet numerical abundance and carbon flux were evident during the spring peak. Combined with data indicating that numerical and carbon fluxes are dominated at 500 m by a distinct type of pellet found uniquely at this depth, these trends suggest the presence of an undescribed mid-water macro-zooplankton or micro-nekton community. Fecal pellet carbon flux was highest at 200 m and varied with depth independently of overall particulate carbon, which was greatest at 500 m. Morphologically distinct types of pellets dominated the numerical and carbon fluxes. Small elliptical and spherical pellets accounted for 88% of the numerical flux, while larger cylindrical pellets, although relatively rare (<10%), accounted for almost 40% of the overall pellet carbon flux. Cylindrical pellets dominated the pellet carbon flux at all depths except 500 m, where a large subtype of elliptical pellet, found only at that depth, was responsible for the majority of pellet carbon flux. Overall during 1990, fecal pellets were responsible for a depth-integrated annual average flux of 1.03 mgC/m**2/d, representing 18% of the total carbon flux. The proportion of vertical carbon flux attributed to fecal pellets varied from 3 to 35%, with higher values occurring during periods when the water column was vertically mixed. Especially during these times, fecal pellets are a critical conveyor of carbon to the deep sea in this region.