First record of microplastics ingestion by European hake MERLUCCIUS MERLUCCIUS from the Tyrrhenian Sicilian coast (Central Mediterranean Sea)

A sample of 67 European hake Merluccius merluccius were examined to highlight the ingestion of microplastics in the Tyrrhenian Sea. In all samples, 31 black fibres were found in the stomach contents corresponding to 46.3% of the specimens. The data presented here could be important for the implementation of the EU Marine Strategy Framework Directive in Mediterranean waters.     

Microparticles Mediate Hepatic Ischemia-Reperfusion Injury and Are the Targets of Diannexin (ASP8597)

Background & Aims

Ischemia–reperfusion injury (IRI) can cause hepatic failure after liver surgery or transplantation. IRI causes oxidative stress, which injures sinusoidal endothelial cells (SECs), leading to recruitment and activation of Kupffer cells, platelets and microcirculatory impairment. We investigated whether injured SECs and other cell types release microparticles during post-ischemic reperfusion, and whether such microparticles have pro-inflammatory, platelet-activating and pro-injurious effects that could contribute to IRI pathogenesis.

Methods

C57BL6 mice underwent 60 min of partial hepatic ischemia followed by 15 min–24 hrs of reperfusion. We collected blood and liver samples, isolated circulating microparticles, and determined protein and lipid content. To establish mechanism for microparticle production, we subjected murine primary hepatocytes to hypoxia-reoxygenation. Because microparticles express everted phosphatidylserine residues that are the target of annexin V, we analyzed the effects of an annexin V-homodimer (Diannexin or ASP8597) on post-ischemia microparticle production and function.

Results

Microparticles were detected in the circulation 15–30 min after post-ischemic reperfusion, and contained markers of SECs, platelets, natural killer T cells, and CD8⁺ cells; 4 hrs later, they contained markers of macrophages. Microparticles contained F2-isoprostanes, indicating oxidative damage to membrane lipids. Injection of mice with TNF-α increased microparticle formation, whereas Diannexin substantially reduced microparticle release and prevented IRI. Hypoxia-re-oxygenation generated microparticles from primary hepatocytes by processes that involved oxidative stress. Exposing cultured hepatocytes to preparations of microparticles isolated from the circulation during IRI caused injury involving mitochondrial membrane permeability transition. Microparticles also activated platelets and induced neutrophil migration in vitro. The inflammatory properties of microparticles involved activation of NF-κB and JNK, increased expression of E-selectin, P-selectin, ICAM-1 and VCAM-1. All these processes were blocked by coating microparticles with Diannexin.

Conclusions

Following hepatic IRI, microparticles circulate and can be taken up by hepatocytes, where they activate signaling pathways that mediate inflammation and hepatocyte injury. Diannexin prevents microparticle formation and subsequent inflammation.     

Quantifying Recent Ecological Changes in Remote Lakes of North America and Greenland Using Sediment Diatom Assemblages

Background

Although arctic lakes have responded sensitively to 20^th-century climate change, it remains uncertain how these ecological transformations compare with alpine and montane-boreal counterparts over the same interval. Furthermore, it is unclear to what degree other forcings, including atmospheric deposition of anthropogenic reactive nitrogen (Nr), have participated in recent regime shifts. Diatom-based paleolimnological syntheses offer an effective tool for retrospective assessments of past and ongoing changes in remote lake ecosystems.

Methodology/Principal Findings

We synthesized 52 dated sediment diatom records from lakes in western North America and west Greenland, spanning broad latitudinal and altitudinal gradients, and representing alpine (n = 15), arctic (n = 20), and forested boreal-montane (n = 17) ecosystems. Diatom compositional turnover (β-diversity) during the 20^th century was estimated using Detrended Canonical Correspondence Analysis (DCCA) for each site and compared, for cores with sufficiently robust chronologies, to both the 19^th century and the prior ∼250 years (Little Ice Age). For both arctic and alpine lakes, β-diversity during the 20^th century is significantly greater than the previous 350 years, and increases with both latitude and altitude. Because no correlation is apparent between 20^th-century diatom β-diversity and any single physical or limnological parameter (including lake and catchment area, maximum depth, pH, conductivity, [NO₃ ⁻], modeled Nr deposition, ambient summer and winter air temperatures, and modeled temperature trends 1948–2008), we used Principal Components Analysis (PCA) to summarize the amplitude of recent changes in relationship to lake pH, lake:catchment area ratio, modeled Nr deposition, and recent temperature trends.

Conclusions/Significance

The ecological responses of remote lakes to post-industrial environmental changes are complex. However, two regions reveal concentrations of sites with elevated 20^th-century diatom β-diversity: the Arctic where temperatures are increasing most rapidly, and mid-latitude alpine lakes impacted by high Nr deposition rates. We predict that remote lakes will continue to shift towards new ecological states in the Anthropocene, particularly in regions where these two forcings begin to intersect geographically.     

Neurokinin 1 Receptor Mediates Membrane Blebbing and Sheer Stress-Induced Microparticle Formation in HEK293 Cells

Cell-derived microparticles participate in intercellular communication similar to the classical messenger systems of small and macro-molecules that bind to specialized membrane receptors. Microparticles have been implicated in the regulation of a variety of complex physiopathologic processes, such as thrombosis, the control of innate and adaptive immunity, and cancer. The neurokinin 1 receptor (NK1R) is a Gq-coupled receptor present on the membrane of a variety of tissues, including neurons in the central and peripheral nervous system, immune cells, endocrine and exocrine glands, and smooth muscle. The endogenous agonist of NK1R is the undecapeptide substance P (SP). We have previously described intracellular signaling mechanisms that regulate NK1R-mediated rapid cell shape changes in HEK293 cells and U373MG cells. In the present study, we show that the activation of NK1R in HEK293 cells, but not in U373MG cells, leads to formation of sheer-stress induced microparticles that stain positive with the membrane-selective fluorescent dye FM 2–10. SP-induced microparticle formation is independent of elevated intracellular calcium concentrations and activation of NK1R present on HEK293-derived microparticles triggers detectable calcium increase in SP-induced microparticles. The ROCK inhibitor Y27632 and the dynamin inhibitor dynasore inhibited membrane blebbing and microparticle formation in HEK293 cells, strongly suggesting that microparticle formation in this cell type is dependent on membrane blebbing.     

Acid sphingomyelinase activity triggers microparticle release from glial cells

We have earlier shown that microglia, the immune cells of the CNS, release microparticles from cell plasma membrane after ATP stimulation. These vesicles contain and release IL-1β, a crucial cytokine in CNS inflammatory events. In this study, we show that microparticles are also released by astrocytes and we get insights into the mechanism of their shedding. We show that, on activation of the ATP receptor P2X₇, microparticle shedding is associated with rapid activation of acid sphingomyelinase, which moves to plasma membrane outer leaflet. ATP-induced shedding and IL-1β release are markedly reduced by the inhibition of acid sphingomyelinase, and completely blocked in glial cultures from acid sphingomyelinase knockout mice. We also show that p38 MAPK cascade is relevant for the whole process, as specific kinase inhibitors strongly reduce acid sphingomyelinase activation, microparticle shedding and IL-1β release. Our results represent the first demonstration that activation of acid sphingomyelinase is necessary and sufficient for microparticle release from glial cells and define key molecular effectors of microparticle formation and IL-1β release, thus, opening new strategies for the treatment of neuroinflammatory diseases.     

Development of a fish-based index (FBI) of biotic integrity for French lakes using the hindcasting approach

The European Water Framework Directive clearly indicates that fish is one of the “quality elements” to be considered for the assessment of lentic systems. However, few fish-based indices (FBIs) of biotic integrity have been developed for lakes so far.Hence, the aim of our study was to develop such a tool for French lakes. Fish surveys, lakes natural environmental parameters, catchment-scale anthropogenic pressures, and local pressures were collected for 67 reservoirs and 24 natural lakes throughout France. After assigning fish species into trophic, reproductive, and tolerance guilds, we derived a set of metrics reflecting complementary aspects of community functioning and condition. Other community-level traits such as richness and evenness were added. These metrics were modeled vs. natural environmental variables and pressures. Reference conditions at each site were then assessed using hindcasting modeling. Separate indices were eventually obtained for natural and artificial lakes by combining selected metrics. Fifteen out of 73 candidate fish metrics, covering all three groups of functional traits, displayed a significant response to anthropogenic pressures. After removal of the redundant traits, the final indices for natural lakes and reservoirs included three and six metrics, respectively. Agricultural-related impacts were prominent for reservoirs, whereas for natural lakes urban and local pressures displayed the most significant effects.     

Land-Use Legacies Are Important Determinants of Lake Eutrophication in the Anthropocene

Background

A hallmark of the latter half of the 20^th century is the widespread, rapid intensification of a variety of anthropogenically-driven environmental changes—a “Great Acceleration.” While there is evidence of a Great Acceleration in a variety of factors known to be linked to water quality degradation, such as conversion of land to agriculture and intensification of fertilizer use, it is not known whether there has been a similar acceleration of freshwater eutrophication.

Methodology/Principal Findings

Using quantitative reconstructions of diatom-inferred total phosphorus (DI-TP) as a proxy for lake trophic state, we synthesized results from 67 paleolimnological studies from across Europe and North America to evaluate whether most lakes showed a pattern of eutrophication with time and whether this trend was accelerated after 1945 CE, indicative of a Great Acceleration. We found that European lakes have experienced widespread increases in DI-TP over the 20^th century and that 33% of these lakes show patterns consistent with a post-1945 CE Great Acceleration. In North America, the proportion of lakes that increased in DI-TP over time is much lower and only 9% exhibited a Great Acceleration of eutrophication.

Conclusions/Significance

The longer and more widespread history of anthropogenic influence in Europe, the leading cause for the relatively pervasive freshwater eutrophication, provides an important cautionary tale; our current path of intensive agriculture around the world may lead to an acceleration of eutrophication in downstream lakes that could take centuries from which to recover.     

Water colour, phosphorus and alkalinity are the major determinants of the dominant phytoplankton species in European lakes

Analysis of phytoplankton data from about 1,500 lakes in 20 European countries has revealed that two-thirds of the species that dominate lakes during the summer are dominant right across Europe. Using Canonical Correspondence Analyses, we have examined how both habitat conditions within lakes and environmental factors over broad geographical scales explained the distribution of the 151 most common summer dominant species. The distributions of these species were best explained by water colour and latitude, although alkalinity and total phosphorus also appeared to be important explanatory factors. Contrary to our original hypothesis, summer water temperatures had a negligible impact on the distribution of dominants, although, due to the restricted summer season we examined, only a limited temperature gradient was present in the dataset. Cryptophytes occurred more frequently among dominants in Northern Europe whereas cyanobacteria and dinophytes dominated more in Central and Southern Europe. Our analyses suggest that besides nutrient concentrations, other water chemistry variables, such as alkalinity and the content of humic substances, have at least as important a role in determining the distribution of the dominant phytoplankton species in European lakes.     

Subfossil Cladocera in relation to contemporary environmental variables in 54 Pan-European lakes

1. Changes in cladoceran subfossils in the surface sediments of 54 shallow lakes were studied along a European latitude gradient (36–68°N). Multivariate methods, such as regression trees and ordination, were applied to explore the relationships between cladoceran taxa distribution and contemporary environmental variables, with special focus on the impact of climate. 2. Multivariate regression tree analysis showed distinct differences in cladoceran community structure and lake characteristics along the latitude gradient, identifying three groups: (i) northern lakes characterised by low annual mean temperature, conductivity, nutrient concentrations and fish abundance, (ii) southern, macrophyte rich, warm water lakes with high conductivity and high fish abundance and (iii) Mid‐European lakes at intermediate latitudes with intermediate conductivities, trophic state and temperatures. 3. Large‐sized, pelagic species dominated a group of seven northern lakes with low conductivity, where acid‐tolerant species were also occasionally abundant. Small‐sized, benthic‐associated species dominated a group of five warm water lakes with high conductivity. Cladoceran communities generally showed low species‐specific preferences for habitat and environmental conditions in the Mid‐European group of lakes. Taxon richness was low in the southern‐most, high‐conductivity lakes as well as in the two northern‐most sub‐arctic lakes. 4. The proportion of cladoceran resting eggs relative to body shields was high in the northern lakes, and linearly (negatively) related to both temperature and Chl a, indicating that both cold climate (short growing season) and low food availability induce high ephippia production. 5. Latitude and, implicitly, temperature were strongly correlated with conductivity and nutrient concentrations, highlighting the difficulties of disentangling a direct climate signal from indirect effects of climate, such as changes in fish community structure and human‐related impacts, when a latitude gradient is used as a climate proxy. Future studies should focus on the interrelationships between latitude and gradients in nutrient concentration and conductivity.     

黄河流域微塑料污染现状及防治策略

微塑料污染(microplastics pollution)在全球范围内受到广泛关注。相比于海洋环境以及其他主要河流、湖泊的微塑料污染情况,黄河流域的相关数据较为贫乏。通过综述文献分析了黄河流域河道沉积物和表层水的微塑料污染丰度、类型以及空间分布特征,探讨了黄河流域重要城市和重点保护区的微塑料污染现状,并提出了相应的防控措施。结果表明：黄河流域沉积物和表层水中微塑料污染在空间分布上整体呈现自上游向下游增多的趋势,尤其在黄河三角洲湿地该趋势更加明显;黄河流域沉积物和表层水中微塑料类型存在明显差异,主要与微塑料的材质有关;与全国同类区域相比,黄河流域国家重点城市市域和国家湿地公园的微塑料污染水平处于中高程度,应引起重视;塑料通过多种暴露途径会对黄河滩区养殖业和人类健康造成严重影响。控制黄河流域水体微塑料污染,需要完善相关生产标准和法律法规,提高可降解微塑料产能和塑料废弃物的工程化降解能力。     

Heterogeneity in Neutrophil Microparticles Reveals Distinct Proteome and Functional Properties

Altered plasma neutrophil microparticle levels have recently been implicated in a number of vascular and inflammatory diseases, yet our understanding of their actions is very limited. Herein, we investigate the proteome of neutrophil microparticles in order to shed light on their biological actions. Stimulation of human neutrophils, either in suspension or adherent to an endothelial monolayer, led to the production of microparticles containing >400 distinct proteins with only 223 being shared by the two subsets. For instance, postadherent microparticles were enriched in alpha-2 macroglobulin and ceruloplasmin, whereas microparticles produced by neutrophils in suspension were abundant in heat shock 70 kDa protein 1. Annexin A1 and lactotransferrin were expressed in both microparticle subsets. We next determined relative abundance of these proteins in three types of human microparticle samples: healthy volunteer plasma, plasma of septic patients and skin blister exudates finding that these proteins were differentially expressed on neutrophil microparticles from these samples reflecting in part the expression profiles we found in vitro. Functional assessment of the neutrophil microparticles subsets demonstrated that in response to direct stimulation neutrophil microparticles produced reactive oxygen species and leukotriene B₄ as well as locomoted toward a chemotactic gradient. Finally, we investigated the actions of the two neutrophil microparticles subsets described herein on target cell responses. Microarray analysis with human primary endothelial cells incubated with either microparticle subset revealed a discrete modulation of endothelial cell gene expression profile. These findings demonstrate that neutrophil microparticles are heterogenous and can deliver packaged information propagating the activation status of the parent cell, potentially exerting novel and fundamental roles both under homeostatic and disease conditions.The emerging notion that cells can communicate by packaged information represents a major shift in our understanding of cell-to-cell interaction in complex settings including inflammation (1). Packaging of mediators (irrespective of their chemical nature) in structures that can be transported through the vascular and lymphatic systems might avoid their rapid dilution and removal by biological fluids and allow the target cell or tissue to receive a biologically relevant amount of a given molecule. As an example, TNF-α produced by mast cells in the mouse paw can reach the lymph nodes unmodified, wrapped up in small structures or vesicles (2). In this respect, the last few years have witnessed augmented understanding in microparticle function.Described over 50 years ago (reviewed in (3, 4), microparticles are heterogeneous in nature with their size varying between 0.2 and 1.0 μm, and are characterized by an outer membrane composed of a phospholipid bilayer and cell surface proteins. The mechanism of microparticle production is not fully understood, though it may follow processes not dissimilar from those observed in apoptosis, involving membrane detachment from the anchoring cytoskeleton and loss of membrane symmetry, which leads to exposure of negatively charged phospholipids (5–7). Proteins found on the outer leaflet of the microparticle cell membrane are believed to reflect both the origin and activation status of the parental cell (8, 9); for instance, microparticles from neutrophils express CD66b and CD62L (10, 11). We have recently identified the selective expression of the potent anti-inflammatory and proresolving protein Annexin A1 (ANXA1) on the surface of microparticles generated from neutrophils adherent to endothelial monolayers, when compared with those prepared from quiescent neutrophils (12). Microparticle production is not restricted to one subset of cells and using cell specific antigens the relative contribution of different cell types to the total microparticle population in a particular environment can be assessed. This has allowed for the analysis of different microparticle populations (the focus being by and large platelet- and endothelial-derived microparticles) in a number of pathologies in the quest to identify robust biomarkers for disease and treatment (13–15). With regard to inflammatory diseases, examples would include plasma samples in sepsis (16), psoriatic arthritis (17), and scleroderma (18). However, the vast majority of these studies have only determined microparticle expression patterns with respect to the cell type of origin, without addressing the possibility that microparticle composition—even when generated from the same leukocyte subset—might differ in relation to disease status and/or mode of cell activation. Of note recent work has also demonstrated that the production of neutrophil microparticles during self-limited inflammation is temporally regulated suggesting that these microparticles are important in orchestrating inflammation-resolution (1).Recent work has established that microparticles can elicit a variety of biological processes ranging from angiogenesis to anti-inflammation; so that it is very unlikely they can continue to be considered “cell debris,” as initially postulated. The following are some examples, relevant to the present study. Ingestion of platelet microparticles alters the phenotype of macrophages, leading to the false identification of endothelial cell progenitor cells in culture (19). Likewise, sonic hedgehog can be transferred, via microparticles, to dysfunctional endothelial cells, restoring the activity of nitric oxide synthase with downstream production of nitric oxide (20). Microparticles can carry functionally active receptor proteins to target cells (21, 22). Finally, in vivo generation of microparticles has been observed within the inflamed microcirculation. Real time analysis of leukocyte recruitment has visualized microparticle release from leukocytes squeezing through an endothelial barrier, providing evidence for their formation in vivo together with potential functional relevance in relation to cell migration (23).On stimulation, neutrophils produce microparticles with rapid and nongenomic anti-inflammatory properties, in vitro and in vivo, reliant on their expression of ANXA1 (12). Whereas these findings are consistent with those obtained by Gasser and colleagues (24) who described inhibitory properties of neutrophil microparticles, other studies have suggested that the same cell type can produce microparticles that elicit activating properties, for instance upon incubation with endothelial cells or monocytes for longer time-points (25, 26). Thus, to gain further insight into the potential mechanisms involved in mediating such distinct effects, we deemed it important to determine the total proteome of neutrophil microparticles. Having established that different stimulation conditions yield microparticle populations with distinct protein profiles, we corroborated our observations in two distinct clinical scenarios, characterizing neutrophil microparticles from skin blister exudates and plasma samples from sepsis patients using a select group of proteins identified in our proteomic profile. We also established that the two microparticles subsets differentially modulate endothelial cell gene expression profile and thereby function, as determined by connectivity map analysis.     

Predicting the spread of invasive species in an uncertain world: accommodating multiple vectors and gaps in temporal and spatial data for <Emphasis Type="Italic">Bythotrephes longimanus</Emphasis>

Real-world uncertainties and data limitations make it difficult to predict how, when and where non-indigenous species (NIS) will spread. Typically only a small fraction of sites are sampled during only a few time intervals, such that we know neither the full spatial extent nor the true temporal progress of invasion. Yet, these unsampled locations might affect the invasion dynamics. We extend propagule pressure models to incorporate both human-mediated and natural fluvial dispersal vectors, and develop techniques to incorporate missing spatial and temporal data on invasions. We apply our model to Bythotrephes longimanus, a high-risk aquatic NIS, using a regional-scale 311-lake survey in a popular watershed in Ontario and extending our analysis to 1,300 unsampled lakes. Of 100 model runs with different random subsets of 50 sampled lakes reserved for validation, we were able to obtain an average area under the curve value of 0.89. Human-mediated dispersal accounted for 99.75% of the contribution of propagules to probability of establishment. Although the discovery rate is accelerating, our results suggest the annual rate of lake invasions is decelerating over time. Management efforts controlling recreational boating traffic out of the largest lakes in the system will be the most effective way of slowing the spread of B. longimanus in lakes within this system.     

Metal accumulation in two contiguous eutrophic peri-urban lakes,Chivero and Manyame,Zimbabwe

Concentrations of aluminium, cadmium, chromium, cobalt, copper, iron, lead, nickel and zinc were determined in surface water, benthic sediments, and the gills, liver and stomach muscle tissues of Oreochromis niloticus and Clarias gariepinus in peri-urban lakes Chivero and Manyame, Zimbabwe. Five sites were sampled in each lake once per month in November 2015, February, May, August and November 2016. Pollution load index detected no metal contamination, whereas the geo-accumulation index reflected heavy to extreme sediment pollution, with Fe, Cd, Zn, Cr, Ni and Cu present in both lakes. Significant spatial temporal variations were detected for Al, Cr, Cu and Pb across sites within and between the two lakes. High Fe, Al and Cr concentrations in water and sediments in lakes Chivero and Manyame derive from geogenic background sources in addition to anthropogenic loads and intensity. Elevated concentrations of Al, Pb, Cu, Cd, Fe and Zn detected in gills, liver and stomach tissue of catfish corroborate concentrations in water and sediments, and pose the highest ecological and health risk for hydrobionts in lakes Chivero and Manyame. Contiguity of peri-urban lakes exposes them to similar threats, necessitating creative water management strategies, which ensure ecological continuity.     

Interaction of P-selectin and PSGL-1 generates microparticles that correct hemostasis in a mouse model of hemophilia A

High plasma levels of soluble P-selectin are associated with thrombotic disorders and may predict future cardiovascular events. Mice with high levels of soluble P-selectin have more microparticles in their plasma than do normal mice. Here we show that chimeras of P-selectin and immunoglobulin (P-sel-Ig) induced formation of procoagulant microparticles in human blood through P-selectin glycoprotein ligand-1 (PSGL-1; encoded by the Psgl1 gene, officially known as Selpl). In addition, Psgl1-/- mice produced fewer microparticles after P-sel-Ig infusion and did not spontaneously increase their microparticle count in old age as do wild-type mice. Injected microparticles specifically bound to thrombi and thus could be involved in thrombin generation at sites of injury. Infusion of P-sel-Ig into hemophilia A mice produced a 20-fold increase over control immunoglobulin in microparticles containing tissue factor. This significantly improved the kinetics of fibrin formation in the hemophilia A mice and normalized their tail-bleeding time. P-sel-Ig treatment could become a new approach to sustained control of bleeding in hemophilia.     

Factors Affecting Elevated Arsenic and Methyl Mercury Concentrations in Small Shield Lakes Surrounding Gold Mines near the Yellowknife,NT, (Canada) Region

Gold mines in the Yellowknife, NT, region—in particular, the Giant Mine—operated from 1949–99, releasing 237,000 tonnes of waste arsenic trioxide (As₂O₃) dust, among other compounds, from gold ore extraction and roasting processes. For the first time, we show the geospatial distribution of roaster-derived emissions of several chemical species beyond the mine property on otherwise undisturbed taiga shield lakes within a 25 km radius of the mine, 11 years after its closing. Additionally, we demonstrate that underlying bedrock is not a significant source for the elevated concentrations in overlying surface waters. Aquatic arsenic (As) concentrations are well above guidelines for drinking water (10 μg/L) and protection for aquatic life (5 μg/L), ranging up to 136 μg/L in lakes within 4 km from the mine, to 2.0 μg/L in lakes 24 km away. High conversion ratios of methyl mercury were shown in lakes near the roaster stack as well, with MeHg concentrations reaching 44% of total mercury. The risk of elevated exposures by these metals is significant, as many lakes used for recreation and fishing near the City of Yellowknife are within this radius of elevated As and methyl Hg concentrations.     

ABCA1-mediated cholesterol efflux generates microparticles in addition to HDL through processes governed by membrane rigidity

ATP-binding cassette transporter A1 (ABCA1) mediates cholesterol efflux to lipid-poor apolipoprotein A-I (apoA-I) and generates HDL. Here, we demonstrate that ABCA1 also directly mediates the production of apoA-I free microparticles. In baby hamster kidney (BHK) cells and RAW macrophages, ABCA1 expression led to lipid efflux in the absence of apoA-I and released large microparticles devoid of apoB and apoE. We provide evidence that these microparticles are an integral component of the classical cholesterol efflux pathway when apoA-I is present and accounted for approximately 30% of the total cholesterol released to the medium. Furthermore, microparticle release required similar ABCA1 activities as was required for HDL production. For instance, a nucleotide binding domain mutation in ABCA1 (A937V) that impaired HDL generation also abolished microparticle release. Similarly, inhibition of protein kinase A (PKA) prevented the release of both types of particles. Interestingly, physical modulation of membrane dynamics affected HDL and microparticle production, rigidifying the plasma membrane with wheat germ agglutinin inhibited HDL and microparticle release, whereas increasing the fluidity promoted the production of these particles. Given the established role of ABCA1 in expending nonraft or more fluid-like membrane domains, our results suggest that both HDL and microparticle release is favored by a more fluid plasma membrane. We speculate that ABCA1 enhances the dynamic movement of the plasma membrane, which is required for apoA-I lipidation and microparticle formation.     

The conservation status of athalassic lakes in New South Wales,Australia

To provide an overview for the State Pollution Control Commission of NSW, 102 lakes were visited throughout the state during the 1988–9 summer to ascertain their ecological condition. The sites chosen covered a spectrum of geomorphic types in approximate proportion to their perceived relative abundance. Field work concentrated on some physicochemical parameters and on zooplankton and littoral invertebrates. A summary of these features of the lakes of NSW is given.The most widespread problem is eutrophication, though for many lakes changes in trophic status could be part of wider changes in lakes since European settlement. A significant number of lakes suffer eroded shorelines and sedimentation. The introduced mosquito fish, Gambusia affinis, is associated with decreased diversity mainly in coastal lakes where also alien plants may be pestiferous. A few lakes in western areas have their flooding regime altered, while a number in the east are drained. With few exceptions there are no management programs to improve the conservation status of degraded lakes in NSW.     

Effects of Microparticle Size and Fc Density on Macrophage Phagocytosis

Controlled induction of phagocytosis in macrophages offers the ability to therapeutically regulate the immune system as well as improve delivery of chemicals or biologicals for immune processing. Maximizing particle uptake by macrophages through Fc receptor-mediated phagocytosis could lead to new delivery mechanisms in drug or vaccine development. Fc ligand density and particle size were examined independently and in combination in order to optimize and tune the phagocytosis of opsonized microparticles. We show the internalization efficiency of small polystyrene particles (0.5 µm to 2 µm) is significantly affected by changes in Fc ligand density, while particles greater than 2 µm show little correlation between internalization and Fc density. We found that while macrophages can efficiently phagocytose a large number of smaller particles, the total volume of phagocytosed particles is maximized through the non-specific uptake of larger microparticles. Therefore, larger microparticles may be more efficient at delivering a greater therapeutic payload to macrophages, but smaller opsonized microparticles can deliver bio-active substances to a greater percentage of the macrophage population. This study is the first to treat as independent variables the physical and biological properties of Fc density and microparticle size that initiate macrophage phagocytosis. Defining the physical and biological parameters that affect phagocytosis efficiency will lead to improved methods of microparticle delivery to macrophages.     

Detection and Quantification of Microparticles from Different Cellular Lineages Using Flow Cytometry. Evaluation of the Impact of Secreted Phospholipase A2 on Microparticle Assessment

Microparticles, also called microvesicles, are submicron extracellular vesicles produced by plasma membrane budding and shedding recognized as key actors in numerous physio(patho)logical processes. Since they can be released by virtually any cell lineages and are retrieved in biological fluids, microparticles appear as potent biomarkers. However, the small dimensions of microparticles and soluble factors present in body fluids can considerably impede their quantification. Here, flow cytometry with improved methodology for microparticle resolution was used to detect microparticles of human and mouse species generated from platelets, red blood cells, endothelial cells, apoptotic thymocytes and cells from the male reproductive tract. A family of soluble proteins, the secreted phospholipases A₂ (sPLA₂), comprises enzymes concomitantly expressed with microparticles in biological fluids and that catalyze the hydrolysis of membrane phospholipids. As sPLA₂ can hydrolyze phosphatidylserine, a phospholipid frequently used to assess microparticles, and might even clear microparticles, we further considered the impact of relevant sPLA2 enzymes, sPLA2 group IIA, V and X, on microparticle quantification. We observed that if enriched in fluids, certain sPLA₂ enzymes impair the quantification of microparticles depending on the species studied, the source of microparticles and the means of detection employed (surface phosphatidylserine or protein antigen detection). This study provides analytical considerations for appropriate interpretation of microparticle cytofluorometric measurements in biological samples containing sPLA2 enzymes.