Investigating Internalization of Reporter-Protein-Functionalized Polyhedrin Particles by Brain Immune Cells.

Achieving sustained drug delivery to the central nervous system (CNS) is a major challenge for neurological injury and disease, and various delivery vehicles are being developed to achieve this. Self-assembling polyhedrin crystals (POlyhedrin Delivery System; PODS) are being exploited for the delivery of therapeutic protein cargo, with demonstrated efficacy in vivo. However, to establish the utility of PODS for neural applications, their handling by neural immune cells (microglia) must be documented, as these cells process and degrade many biomaterials, often preventing therapeutic efficacy. Here, primary mouse cortical microglia were cultured with a GFP-functionalized PODS for 24 h. Cell counts, cell morphology and Iba1 expression were all unaltered in treated cultures, indicating a lack of acute toxicity or microglial activation. Microglia exhibited internalisation of the PODS, with both cytosolic and perinuclear localisation. No evidence of adverse effects on cellular morphology was observed. Overall, 20-40% of microglia exhibited uptake of the PODS, but extracellular/non-internalised PODS were routinely present after 24 h, suggesting that extracellular drug delivery may persist for at least 24 h.

Impacts of artificial light at night on the early life history of two ecosystem engineers.

Sessile marine invertebrates play a vital role as ecosystem engineers and in benthic-pelagic coupling. Most benthic fauna develop through larval stages and the importance of natural light cycles for larval biology and ecology is long-established. Natural light-dark cycles regulate two of the largest ocean-scale processes that are fundamental to larvae's life cycle: the timing of broadcast spawning for successful fertilization and diel vertical migration for foraging and predator avoidance. Given the reliance on light and the ecological role of larvae, surprisingly little is known about the impacts of artificial light at night (ALAN) on the early life history of habitat-forming species. We quantified ALAN impacts on larval performance (survival, growth, development) of two cosmopolitan ecosystem engineers in temperate marine ecosystems, the mussel Mytilus edulis and the barnacle Austrominius modestus. Higher ALAN irradiance reduced survival in both species (57% and 13%, respectively). ALAN effects on development and growth were small overall, and different between species, time-points and parentage. Our results show that ALAN adversely affects larval survival and reiterates the importance of paternal influence on offspring performance. ALAN impacts on the early life stages of ecosystem engineering species have implications not only for population viability but also the ecological communities that these species support. This article is part of the theme issue 'Light pollution in complex ecological systems'.

Top ten priorities for global saltmarsh restoration, conservation and ecosystem service research.

Coastal saltmarshes provide globally important ecosystem services including 'blue carbon' sequestration, flood protection, pollutant remediation, habitat provision and cultural value. Large portions of marshes have been lost or fragmented as a result of land reclamation, embankment construction, and pollution. Sea level rise threatens marsh survival by blocking landward migration where coastlines have been developed. Research-informed saltmarsh conservation and restoration efforts are helping to prevent further loss, yet significant knowledge gaps remain. Using a mixed methods approach, this paper identifies ten research priorities through an online questionnaire and a residential workshop attended by an international, multi-disciplinary network of 35 saltmarsh experts spanning natural, physical and social sciences across research, policy, and practitioner sectors. Priorities have been grouped under four thematic areas of research: Saltmarsh Area Extent, Change and Restoration Potential (including past, present, global variation), Spatio-social contexts of Ecosystem Service delivery (e.g. influences of environmental context, climate change, and stakeholder groups on service provisioning), Patterns and Processes in saltmarsh functioning (global drivers of saltmarsh ecosystem structure/function) and Management and Policy Needs (how management varies contextually; challenges/opportunities for management). Although not intended to be exhaustive, the challenges, opportunities, and strategies for addressing each research priority examined here, providing a blueprint of the work that needs to be done to protect saltmarshes for future generations.

A requirement for astrocyte IP3R2 signaling for whisker experience-dependent depression and homeostatic upregulation in the mouse barrel cortex.

Changes to sensory experience result in plasticity of synapses in the cortex. This experience-dependent plasticity (EDP) is a fundamental property of the brain. Yet, while much is known about neuronal roles in EDP, very little is known about the role of astrocytes. To address this issue, we used the well-described mouse whiskers-to-barrel cortex system, which expresses a number of forms of EDP. We found that all-whisker deprivation induced characteristic experience-dependent Hebbian depression (EDHD) followed by homeostatic upregulation in L2/3 barrel cortex of wild type mice. However, these changes were not seen in mutant animals (IP3R2-/-) that lack the astrocyte-expressed IP3 receptor subtype. A separate paradigm, the single-whisker experience, induced potentiation of whisker-induced response in both wild-type (WT) mice and IP3R2-/- mice. Recordings in ex vivo barrel cortex slices reflected the in vivo results so that long-term depression (LTD) could not be elicited in slices from IP3R2-/- mice, but long-term potentiation (LTP) could. Interestingly, 1 Hz stimulation inducing LTD in WT paradoxically resulted in NMDAR-dependent LTP in slices from IP3R2-/- animals. The LTD to LTP switch was mimicked by acute buffering astrocytic [Ca2+] i in WT slices. Both WT LTD and IP3R2-/- 1 Hz LTP were mediated by non-ionotropic NMDAR signaling, but only WT LTD was P38 MAPK dependent, indicating an underlying mechanistic switch. These results demonstrate a critical role for astrocytic [Ca2+] i in several EDP mechanisms in neocortex.

Artificial light at night reverses monthly foraging pattern under simulated moonlight.

Mounting evidence shows that artificial light at night (ALAN) alters biological processes across levels of organization, from cells to communities. Yet, the combined impacts of ALAN and natural sources of night-time illumination remain little explored. This is in part due the lack of accurate simulations of the complex changes moonlight intensity, timing and spectra throughout a single night and lunar cycles in laboratory experiments. We custom-built a novel system to simulate natural patterns of moonlight to test how different ALAN intensities affect predator-prey relationships over the full lunar cycle. Exposure to high intensity ALAN (10 and 50 lx) reversed the natural lunar-guided foraging pattern by the gastropod mesopredator Nucella lapillus on its prey Semibalanus balanoides. Foraging decreased during brighter moonlight in naturally lit conditions. When exposed to high intensity ALAN, foraging increased with brighter moonlight. Low intensity ALAN (0.1 and 0.5 lx) had no impact on foraging. Our results show that ALAN alters the foraging pattern guided by changes in moonlight brightness. ALAN impacts on ecosystems can depend on lunar light cycles. Accurate simulations of night-time light cycle will warrant more realistic insights into ALAN impacts and also facilitate advances in fundamental night-time ecology and chronobiology.

Local and large-scale spatial variation in a marine predator-prey interaction in the southwestern Atlantic.

Predator-prey interactions are a key ecological process which can be modified by environmental conditions over a range of spatial scales. Through two complementary short-term experiments, we assessed how local and large-scale environmental conditions affect a subtropical intertidal predator-prey interaction. At a local scale, we evaluated the effects of the degree of exposure to wave action and prey density on consumption rate and interaction strength using a whelk-barnacle system. Consumption rate decreased with wave exposure at experimentally reduced prey density but did not change at ambient density. Such an interactive effect occurred due to shifts in the whelk's feeding behaviour, likely linked to encounter rate and stress amelioration underpinned by prey density. Per capita interaction strength of the whelk on the barnacle weakened along the wave exposure gradient, but to a greater degree at reduced compared to ambient prey density. This confirms that environmental harshness can decrease the importance of predators, but the magnitude of change may be modified by density-dependent effects. A large-scale experiment did not reveal spatial patterns in the whelk-barnacle interaction, nor relationships to chlorophyll-a concentration or the minor change in sea temperature across the study area. Patterns in the size of consumed barnacles along the chlorophyll-a gradient suggest changes in food choice related to prey quality and size. We conclude that disentangling the effects of wave exposure and prey density revealed important potential mechanisms driving species locally. Large-scale variation in the whelk-barnacle interaction appeared to be linked to species' traits shaped by the environmental context.

When One Health Meets the United Nations Ocean Decade: Global Agendas as a Pathway to Promote Collaborative Interdisciplinary Research on Human-Nature Relationships.

Strong evidence shows that exposure and engagement with the natural world not only improve human wellbeing but can also help promote environmentally friendly behaviors. Human-nature relationships are at the heart of global agendas promoted by international organizations including the World Health Organization's (WHO) "One Health" and the United Nations (UN) "Ocean Decade." These agendas demand collaborative multisector interdisciplinary efforts at local, national, and global levels. However, while global agendas highlight global goals for a sustainable world, developing science that directly addresses these agendas from design through to delivery and outputs does not come without its challenges. In this article, we present the outcomes of international meetings between researchers, stakeholders, and policymakers from the United Kingdom and Brazil. We propose a model for interdisciplinary work under such global agendas, particularly the interface between One Health and the UN Ocean Decade and identify three priority research areas closely linked to each other: human-nature connection, conservation-human behavior, and implementation strategies (bringing stakeholders together). We also discuss a number of recommendations for moving forward.

Quantification of blue carbon pathways contributing to negative feedback on climate change following glacier retreat in West Antarctic fjords.

Global warming is causing significant losses of marine ice around the polar regions. In Antarctica, the retreat of tidewater glaciers is opening up novel, low-energy habitats (fjords) that have the potential to provide a negative feedback loop to climate change. These fjords are being colonized by organisms on and within the sediment and act as a sink for particulate matter. So far, blue carbon potential in Antarctic habitats has mainly been estimated using epifaunal megazoobenthos (although some studies have also considered macrozoobenthos). We investigated two further pathways of carbon storage and potential sequestration by measuring the concentration of carbon of infaunal macrozoobenthos and total organic carbon (TOC) deposited in the sediment. We took samples along a temporal gradient since time of last glacier ice cover (1-1000 years) at three fjords along the West Antarctic Peninsula. We tested the hypothesis that seabed carbon standing stock would be mainly driven by time since last glacier covered. However, results showed this to be much more complex. Infauna were highly variable over this temporal gradient and showed similar total mass of carbon standing stock per m2 as literature estimates of Antarctic epifauna. TOC mass in the sediment, however, was an order of magnitude greater than stocks of infaunal and epifaunal carbon and increased with time since last ice cover. Thus, blue carbon stocks and recent gains around Antarctica are likely much higher than previously estimated as is their negative feedback on climate change.

Methane and the Paris Agreement temperature goals.

Meeting the Paris Agreement temperature goal necessitates limiting methane (CH4)-induced warming, in addition to achieving net-zero or (net-negative) carbon dioxide (CO2) emissions. In our model, for the median 1.5°C scenario between 2020 and 2050, CH4 mitigation lowers temperatures by 0.1°C; CO2 increases it by 0.2°C. CO2 emissions continue increasing global mean temperature until net-zero emissions are reached, with potential for lowering temperatures with net-negative emissions. By contrast, reducing CH4 emissions starts to reverse CH4-induced warming within a few decades. These differences are hidden when framing climate mitigation using annual 'CO2-equivalent' emissions, including targets based on aggregated annual emission rates. We show how the different warming responses to CO2 and CH4 emissions can be accurately aggregated to estimate warming by using 'warming-equivalent emissions', which provide a transparent and convenient method to inform policies and measures for mitigation, or demonstrate progress towards a temperature goal. The method presented (GWP*) uses well-established climate science concepts to relate GWP100 to temperature, as a simple proxy for a climate model. The use of warming-equivalent emissions for nationally determined contributions and long-term strategies would enhance the transparency of stocktakes of progress towards a long-term temperature goal, compared to the use of standard equivalence methods. This article is part of a discussion meeting issue 'Rising methane: is warming feeding warming? (part 2)'.

In vitro model of traumatic brain injury to screen neuro-regenerative biomaterials.

Penetrating traumatic brain injury (pTBI) causes serious neurological deficits with no clinical regenerative therapies currently available. Tissue engineering strategies using biomaterial-based 'structural bridges' offer high potential to promote neural regeneration post-injury. This includes surgical grade materials which can be repurposed as biological scaffolds to overcome challenges associated with long approval processes and scaleup for human application. However, high throughput, pathomimetic models of pTBI are lacking for the developmental testing of such neuro-materials, representing a bottleneck in this rapidly emergent field. We have established a high throughput and facile culture model containing the major neural cell types which govern biomaterial handling in the central nervous system. We show that induction of traumatic injuries was feasible in the model, with post-injury implantation of a surgical grade biomaterial. Cellular imaging in lesions was achievable using standard epifluorescence microscopy methods. Key pathological features of pTBI were evident in vitro namely immune cell infiltration of lesions/biomaterial, with responses characteristic of cell scarring, namely hypertrophic astrocytes with GFAP upregulation. Based on our observations, we consider the high-throughput, inexpensive and facile pTBI model can be used to study biomaterial 'implantation' and evaluate neural cell-biomaterial responses. The model is highly versatile to test a range of laboratory and clinical grade materials for neural regeneration.

Influence of environmental variables over multiple spatial scales on the population structure of a key marine invertebrate.

Quantifying scale-dependent patterns and linking ecological to environmental variation is required to understand mechanisms regulating biodiversity. We conducted a large-scale survey in rocky shores along the SE Brazilian coast to examine spatial variability in body size and density of an intertidal barnacle (Chthamalus bisinuatus) and its relationships with benthic and oceanographic predictors. Both the size and density of barnacles showed most variation at the smallest spatial scales. On average, barnacle body size was larger on shores located in areas characterised by higher chlorophyll levels, colder waters, low wave action and low influence of freshwater. Barnacles were more abundant at wave-exposed shores. We identified critical scales of spatial variation of an important species and linked population patterns to essential environmental predictors. Our results show that populations of this barnacle are coupled to scale-dependent oceanographic variation. This study offers insights into the mechanisms regulating coastal populations along a little studied coastline.

Ensuring that offsets and other internationally transferred mitigation outcomes contribute effectively to limiting global warming.

Ensuring the environmental integrity of internationally transferred mitigation outcomes, whether through offset arrangements, a market mechanism or non-market approaches, is a priority for the implementation of Article 6 of the Paris Agreement. Any conventional transferred mitigation outcome, such as an offset agreement, that involves exchanging greenhouse gases with different lifetimes can increase global warming on some timescales. We show that a simple 'do no harm' principle regarding the choice of metrics to use in such transactions can be used to guard against this, noting that it may also be applicable in other contexts such as voluntary and compliance carbon markets. We also show that both approximate and exact 'warming equivalent' exchanges are possible, but present challenges of implementation in any conventional market. Warming-equivalent emissions may, however, be useful in formulating warming budgets in a two-basket approach to mitigation and in reporting contributions to warming in the context of the global stocktake.

Artificial shorelines lack natural structural complexity across scales.

From microbes to humans, habitat structural complexity plays a direct role in the provision of physical living space, and increased complexity supports higher biodiversity and ecosystem functioning across biomes. Coastal development and the construction of artificial shorelines are altering natural landscapes as humans seek socio-economic benefits and protection from coastal storms, flooding and erosion. In this study, we evaluate how much structural complexity is missing on artificial coastal structures compared to natural rocky shorelines, across a range of spatial scales from 1 mm to 10 s of m, using three remote sensing platforms (handheld camera, terrestrial laser scanner and uncrewed aerial vehicles). Natural shorelines were typically more structurally complex than artificial ones and offered greater variation between locations. However, our results varied depending on the type of artificial structure and the scale at which complexity was measured. Seawalls were deficient at all scales (approx. 20-40% less complex than natural shores), whereas rock armour was deficient at the smallest and largest scales (approx. 20-50%). Our findings reinforce concerns that hardening shorelines with artificial structures simplifies coastlines at organism-relevant scales. Furthermore, we offer much-needed insight into how structures might be modified to more closely capture the complexity of natural rocky shores that support biodiversity.

Astrocyte Activation in Neurovascular Damage and Repair Following Ischaemic Stroke.

Transient or permanent loss of tissue perfusion due to ischaemic stroke can lead to damage to the neurovasculature, and disrupt brain homeostasis, causing long-term motor and cognitive deficits. Despite promising pre-clinical studies, clinically approved neuroprotective therapies are lacking. Most studies have focused on neurons while ignoring the important roles of other cells of the neurovascular unit, such as astrocytes and pericytes. Astrocytes are important for the development and maintenance of the blood-brain barrier, brain homeostasis, structural support, control of cerebral blood flow and secretion of neuroprotective factors. Emerging data suggest that astrocyte activation exerts both beneficial and detrimental effects following ischaemic stroke. Activated astrocytes provide neuroprotection and contribute to neurorestoration, but also secrete inflammatory modulators, leading to aggravation of the ischaemic lesion. Astrocytes are more resistant than other cell types to stroke pathology, and exert a regulative effect in response to ischaemia. These roles of astrocytes following ischaemic stroke remain incompletely understood, though they represent an appealing target for neurovascular protection following stroke. In this review, we summarise the astrocytic contributions to neurovascular damage and repair following ischaemic stroke, and explore mechanisms of neuroprotection that promote revascularisation and neurorestoration, which may be targeted for developing novel therapies for ischaemic stroke.

Larval tolerance to food limitation is stronger in an exotic barnacle than in its native competitor.

A critical question in marine ecology is understanding how organisms will cope with environmental conditions under climate change. Increasing temperatures not only have a direct effect on marine organisms but may also lead to food limitation through for example trophic mismatches, or by the increased metabolic demands imposed by developing at high temperatures. Using barnacles from a population of North Wales, we studied the combined effect of temperature and food density on the survival, settlement success, developmental time and body size of larvae of the native barnacle Semibalanus balanoides and its exotic competitor, the barnacle Austrominius modestus. Larvae were reared at similar food levels but at temperature ranges which varied among species reflecting their different phenology and tolerances. For S. balanoides (spring larval release) we used a lower temperature of 9 °C, reflecting spring temperatures from N Wales to SW England, and 15 °C representing warmer conditions; for A. modestus (summer larval release) a typical summer temperature for this geographic range of 15 °C was used with a raised temperature of 18 °C. Larvae were reared under controlled conditions in automated, computer programmable incubators and fed diatoms (Skeletonema costatum) at three food levels. We found stress effects of food limitation on larval performance of S. balanoides. While survival during naupliar development was little affected by food and temperature, low food levels strongly depressed survival and settlement during the cyprid stage of S. balanoides at both tested temperatures, but especially at 15 °C. By contrast, at the tested temperatures little effects were found on survival and settlement success in the exotic A. modestus. Both species delayed development in response to low food levels while S. balanoides cyprids showed decreased body size at the high tested temperature. The main impact occurred as a delayed effect, at the time when cyprids attempt to settle, rather than as an effect on naupliar survival or metamorphosis to the cyprid stage. Response in body size and developmental time may have costs at the time of metamorphosis (delayed settlement) or after metamorphosis. Overall, our experiments suggest that as temperature increases, settlement success of S. balanoides larvae (but not that of its competitor A. modestus) will become more sensitive to conditions of food limitation, imposed for instance by phenological mismatches with periods of phytoplankton peak.

Impacts of copper contamination on a rocky intertidal predator-prey interaction.

Metal contamination can change ecological interactions with potential effects on community dynamics. However, understanding real effects of metals on biota relies on studies undertaken in natural conditions. Through a field experiment, we investigated the effects of copper contamination on the responses of a barnacle prey and its predator, the dogwhelk, and explicitly their interaction. Contamination increased barnacle mortality and reduced predation with no effects on interaction strength. This was because the higher mortality of the prey compensated for the lower consumption of the predator. Despite not affecting the interaction strength, these results suggest a decrease in energy flow in the trophic chain that may lead to important changes in community structure and ecosystem functioning. This study shows the importance of manipulative experiments designed to provide mechanistic insights into ecological interactions to better clarify the effect of stressors on the structure and dynamic of communities.

Metal pollution as a potential threat to shell strength and survival in marine bivalves.

Marine bivalve molluscs, such as scallops, mussels and oysters, are crucial components of coastal ecosystems, providing a range of ecosystem services, including a quarter of the world's seafood. Unfortunately, coastal marine areas often suffer from high levels of metals due to dumping and disturbance of contaminated material. We established that increased levels of metal pollution (zinc, copper and lead) in sediments near the Isle of Man, resulting from historical mining, strongly correlated with significant weakening of shell strength in king scallops, Pecten maximus. This weakness increased mortality during fishing and left individuals more exposed to predation. Comparative structural analysis revealed that shells from the contaminated area were thinner and exhibited a pronounced mineralisation disruption parallel to the shell surface within the foliated region of both the top and bottom valves. Our data suggest that these disruptions caused reduced fracture strength and hence increased mortality, even at subcritical contamination levels with respect to current international standards. This hitherto unreported effect is important since such non-apical responses rarely feed into environmental quality assessments, despite potentially significant implications for the survival of organisms exposed to contaminants. Hence our findings highlight the impact of metal pollution on shell mineralisation in bivalves and urge a reappraisal of currently accepted critical contamination levels.