Integrin-linked kinase regulates the rate of platelet activation and is essential for the formation of stable thrombi

BACKGROUND: Integrin-linked kinase (ILK) and its associated complex of proteins are involved in many cellular activation processes, including cell adhesion and integrin signaling. We have previously demonstrated that mice with induced platelet ILK deficiency show reduced platelet activation and aggregation, but only a minor bleeding defect. Here, we explore this apparent disparity between the cellular and hemostatic phenotypes. METHODS: The impact of ILK inhibition on integrin αII b β3 activation and degranulation was assessed with the ILK-specific inhibitor QLT0267, and a conditional ILK-deficient mouse model was used to assess the impact of ILK deficiency on in vivo platelet aggregation and thrombus formation. RESULTS: Inhibition of ILK reduced the rate of both fibrinogen binding and α-granule secretion, but was accompanied by only a moderate reduction in the maximum extent of platelet activation or aggregation in vitro. The reduction in the rate of fibrinogen binding occurred prior to degranulation or translocation of αII b β3 to the platelet surface. The change in the rate of platelet activation in the absence of functional ILK led to a reduction in platelet aggregation in vivo, but did not change the size of thrombi formed following laser injury of the cremaster arteriole wall in ILK-deficient mice. It did, however, result in a marked decrease in the stability of thrombi formed in ILK-deficient mice. CONCLUSION: Taken together, the findings of this study indicate that, although ILK is not essential for platelet activation, it plays a critical role in facilitating rapid platelet activation, which is essential for stable thrombus formation.

Interactive effect of floral abundance and semi-natural habitats on pollinators in field beans (Vicia faba)

Pollination services are economically important component of agricultural biodiversity which enhance the yield and quality of many crops. An understanding of the suitability of extant habitats for pollinating species is crucial for planning management actions to protect and manage these service providers. In a highly modified agricultural ecosystem, we tested the effect of different pollination treatments (open, autonomous self- and wind-pollination) on pod set, seed set, and seed weight in field beans (Vicia faba). We also investigated the effect of semi-natural habitats and flower abundance on pollinators of field beans. Pollinator sampling was undertaken in ten field bean fields along a gradient of habitat complexity; CORINE land cover classification was used to analyse the land use patterns between 500–3000 m around the sites. Total yield from open-pollination increased by 185% compared to autonomous self-pollination. There was positive interactive effect of local flower abundance and cover of semi-natural habitats on overall abundance of pollinators at 1500 and 2000 m, and abundance of bumblebees (Bombus spp.) at 1000–2000 m. In contrast, species richness of pollinators was only correlated with flower abundance and not with semi-natural habitats. We did not find a link between pod set from open-pollination and pollinator abundance, possibly due to variations in the growing conditions and pollinator communities between sites. We conclude that insect pollination is essential for optimal bean yields and therefore the maintenance of semi-natural habitats in agriculture-dominated landscapes should ensure stable and more efficient pollination services in field beans.

The impact of over 80 years of land cover changes on bee and wasp pollinator communities in England

Change in land cover is thought to be one of the key drivers of pollinator declines, and yet there is a dearth of studies exploring the relationships between historical changes in land cover and shifts in pollinator communities. Here, we explore, for the first time, land cover changes in England over more than 80 years, and relate them to concurrent shifts in bee and wasp species richness and community composition. Using historical data from 14 sites across four counties, we quantify the key land cover changes within and around these sites and estimate the changes in richness and composition of pollinators. Land cover changes within sites, as well as changes within a 1 km radius outside the sites, have significant effects on richness and composition of bee and wasp species, with changes in edge habitats between major land classes also having a key influence. Our results highlight not just the land cover changes that may be detrimental to pollinator communities, but also provide an insight into how increases in habitat diversity may benefit species diversity, and could thus help inform policy and practice for future land management.

Quantifying the relative importance of land cover change from climate and land-use in the representative concentration pathways

Climate change is projected to cause substantial alterations in vegetation distribution, but these have been given little attention in comparison to land-use in the Representative Concentration Pathway (RCP) scenarios. Here we assess the climate-induced land cover changes (CILCC) in the RCPs, and compare them to land-use land cover change (LULCC). To do this, we use an ensemble of simulations with and without LULCC in earth system model HadGEM2-ES for RCP2.6, RCP4.5 and RCP8.5. We find that climate change causes an expansion poleward of vegetation that affects more land area than LULCC in all of the RCPs considered here. The terrestrial carbon changes from CILCC are also larger than for LULCC. When considering only forest, the LULCC is larger, but the CILCC is highly variable with the overall radiative forcing of the scenario. The CILCC forest increase compensates 90% of the global anthropogenic deforestation by 2100 in RCP8.5, but just 3% in RCP2.6. Overall, bigger land cover changes tend to originate from LULCC in the shorter term or lower radiative forcing scenarios, and from CILCC in the longer term and higher radiative forcing scenarios. The extent to which CILCC could compensate for LULCC raises difficult questions regarding global forest and biodiversity offsetting, especially at different timescales. This research shows the importance of considering the relative size of CILCC to LULCC, especially with regard to the ecological effects of the different RCPs.

Nicked-site substrates for a serine recombinase reveal enzyme–DNA communications and an essential tethering role of covalent enzyme–DNA linkages

To analyse the mechanism and kinetics of DNA strand cleavages catalysed by the serine recombinase Tn3 resolvase, we made modified recombination sites with a single-strand nick in one of the two DNA strands. Resolvase acting on these sites cleaves the intact strand very rapidly, giving an abnormal half-site product which accumulates. We propose that these reactions mimic second-strand cleavage of an unmodified site. Cleavage occurs in a synapse of two sites, held together by a resolvase tetramer; cleavage at one site stimulates cleavage at the partner site. After cleavage of a nicked-site substrate, the half-site that is not covalently linked to a resolvase subunit dissociates rapidly from the synapse, destabilizing the entire complex. The covalent resolvase–DNA linkages in the natural reaction intermediate thus perform an essential DNA-tethering function. Chemical modifications of a nicked-site substrate at the positions of the scissile phosphodiesters result in abolition or inhibition of resolvase-mediated cleavage and effects on resolvase binding and synapsis, providing insight into the serine recombinase catalytic mechanism and how resolvase interacts with the substrate DNA.

A framework for designing multi-functional cover crops

Cover crops are sown to provide a number of ecosystem services including nutrient management, mitigation of diffuse pollution, improving soil structure and organic matter content, weed suppression, nitrogen fixation and provision of resources for biodiversity. Although the decision to sow a cover crop may be driven by a desire to achieve just one of these objectives, the diversity of cover crops species and mixtures available means that there is potential to combine a number of ecosystem services within the same crop and growing season. Designing multi-functional cover crops would potentially help to reconcile the often conflicting agronomic and environmental agendas and contribute to the optimal use of land. We present a framework for integrating multiple ecosystem services delivered by cover crops that aims to design a mixture of species with complementary growth habit and functionality. The optimal number and identity of species will depend on the services included in the analysis, the functional space represented by the available species pool and the community dynamics of the crop in terms of dominance and co-existence. Experience from a project that applied the framework to fertility building leys in organic systems demonstrated its potential and emphasised the importance of the initial choice of species to include in the analysis

CLEC-2 and Syk in the megakaryocytic/platelet lineage are essential for development

The C-type lectin receptor CLEC-2 signals through a pathway that is critically dependent on the tyrosine kinase Syk. We show that homozygous loss of either protein results in defects in brain vascular and lymphatic development, lung inflation, and perinatal lethality. Furthermore, we find that conditional deletion of Syk in the hematopoietic lineage, or conditional deletion of CLEC-2 or Syk in the megakaryocyte/platelet lineage, also causes defects in brain vascular and lymphatic development, although the mice are viable. In contrast, conditional deletion of Syk in other hematopoietic lineages had no effect on viability or brain vasculature and lymphatic development. We show that platelets, but not platelet releasate, modulate the migration and intercellular adhesion of lymphatic endothelial cells through a pathway that depends on CLEC-2 and Syk. These studies found that megakaryocyte/platelet expression of CLEC-2 and Syk is required for normal brain vasculature and lymphatic development and that platelet CLEC-2 and Syk directly modulate lymphatic endothelial cell behavior in vitro.

MiR-375 is essential for human spinal motor neuron development and may be involved in motor neuron degeneration

The transcription factor REST is a key suppressor of neuronal genes in non-neuronal tissues. REST has been shown to suppress pro-neuronal microRNAs in neural progenitors indicating that REST-mediated neurogenic suppression may act in part via microRNAs. We used neural differentiation of Rest-null mouse ESC to identify dozens of microRNAs regulated by REST during neural development. One of the identified microRNAs, miR-375, was upregulated during human spinal motor neuron development. We found that miR-375 facilitates spinal motor neurogenesis by targeting the cyclin kinase CCND2 and the transcription factor PAX6. Additionally, miR-375 inhibits the tumor suppressor p53 and protects neurons from apoptosis in response to DNA damage. Interestingly, motor neurons derived from a spinal muscular atrophy patient displayed depressed miR-375 expression and elevated p53 protein levels. Importantly, SMA motor neurons were significantly more susceptible to DNA damage induced apoptosis suggesting that miR-375 may play a protective role in motor neurons.

Scale interactions between the MJO and the western maritime continent

State-of-the-art regional climate model simulations that are able to resolve key mesoscale circulations are used, for the first time, to understand the interaction between the large-scale convective environment of the MJO and processes governing the strong diurnal cycle over the islands of the Maritime Continent (MC). Convection is sustained in the late afternoon just inland of the coasts due to sea breeze convergence. Previous work has shown that the variability in MC rainfall associated with the MJO is manifested in changes to this diurnal cycle; land-based rainfall peaks before the active convective envelope of the MJO reaches the MC, whereas oceanic rainfall rates peak whilst the active envelope resides over the region. The model simulations show that the main controls on oceanic MC rainfall in the early active MJO phases are the large-scale environment and atmospheric stability, followed by high oceanic latent heat flux forced by high near-surface winds in the later active MJO phases. Over land, rainfall peaks before the main convective envelope arrives (in agreement with observations), even though the large-scale convective environment is only moderately favourable for convection. The causes of this early rainfall peak are convective triggers from land-sea breeze circulations that are strong due to high surface insolation and surface heating. During the peak MJO phases cloud cover increases and surface insolation decreases, which weakens the strength of the mesoscale circulations and reduces land-based rainfall, even though the large-scale environment remains favourable for convection at this time. Hence, scale interactions are an essential part of the MJO transition across the MC.

Intercomparison of the Arctic sea ice cover in global ocean–sea ice reanalyses from the ORA-IP project

Ocean–sea ice reanalyses are crucial for assessing the variability and recent trends in the Arctic sea ice cover. This is especially true for sea ice volume, as long-term and large scale sea ice thickness observations are inexistent. Results from the Ocean ReAnalyses Intercomparison Project (ORA-IP) are presented, with a focus on Arctic sea ice fields reconstructed by state-of-the-art global ocean reanalyses. Differences between the various reanalyses are explored in terms of the effects of data assimilation, model physics and atmospheric forcing on properties of the sea ice cover, including concentration, thickness, velocity and snow. Amongst the 14 reanalyses studied here, 9 assimilate sea ice concentration, and none assimilate sea ice thickness data. The comparison reveals an overall agreement in the reconstructed concentration fields, mainly because of the constraints in surface temperature imposed by direct assimilation of ocean observations, prescribed or assimilated atmospheric forcing and assimilation of sea ice concentration. However, some spread still exists amongst the reanalyses, due to a variety of factors. In particular, a large spread in sea ice thickness is found within the ensemble of reanalyses, partially caused by the biases inherited from their sea ice model components. Biases are also affected by the assimilation of sea ice concentration and the treatment of sea ice thickness in the data assimilation process. An important outcome of this study is that the spatial distribution of ice volume varies widely between products, with no reanalysis standing out as clearly superior as compared to altimetry estimates. The ice thickness from systems without assimilation of sea ice concentration is not worse than that from systems constrained with sea ice observations. An evaluation of the sea ice velocity fields reveals that ice drifts too fast in most systems. As an ensemble, the ORA-IP reanalyses capture trends in Arctic sea ice area and extent relatively well. However, the ensemble can not be used to get a robust estimate of recent trends in the Arctic sea ice volume. Biases in the reanalyses certainly impact the simulated air–sea fluxes in the polar regions, and questions the suitability of current sea ice reanalyses to initialize seasonal forecasts.

Cloud information content analysis of multi-angular measurements in the oxygen A-band: application to 3MI and MSPI

The vertical distribution of cloud cover has a significant impact on a large number of meteorological and climatic processes. Cloud top altitude and cloud geometrical thickness are then essential. Previous studies established the possibility of retrieving those parameters from multi-angular oxygen A-band measurements. Here we perform a study and comparison of the performances of future instruments. The 3MI (Multi-angle, Multi-channel and Multi-polarization Imager) instrument developed by EUMETSAT, which is an extension of the POLDER/PARASOL instrument, and MSPI (Multi-angles Spectro-Polarimetric Imager) develoloped by NASA's Jet Propulsion Laboratory will measure total and polarized light reflected by the Earth's atmosphere–surface system in several spectral bands (from UV to SWIR) and several viewing geometries. Those instruments should provide opportunities to observe the links between the cloud structures and the anisotropy of the reflected solar radiation into space. Specific algorithms will need be developed in order to take advantage of the new capabilities of this instrument. However, prior to this effort, we need to understand, through a theoretical Shannon information content analysis, the limits and advantages of these new instruments for retrieving liquid and ice cloud properties, and especially, in this study, the amount of information coming from the A-Band channel on the cloud top altitude (CTOP) and geometrical thickness (CGT). We compare the information content of 3MI A-Band in two configurations and that of MSPI. Quantitative information content estimates show that the retrieval of CTOP with a high accuracy is possible in almost all cases investigated. The retrieval of CGT seems less easy but possible for optically thick clouds above a black surface, at least when CGT > 1–2 km.