Itaconic acid indicates cellular but not systemic immune system activation.

Itaconic acid is produced by mammalian leukocytes upon pro-inflammatory activation. It appears to inhibit bacterial growth and to rewire the metabolism of the host cell by inhibiting succinate dehydrogenase. Yet, it is unknown whether itaconic acid acts only intracellularly, locally in a paracrine fashion, or whether it is even secreted from the inflammatory cells at meaningful levels in peripheral blood of patients with severe inflammation or sepsis. The aim of this study was to determine the release rate of itaconic acid from pro-inflammatory activated macrophages in vitro and to test for the abundance of itaconic acid in bodyfluids of patients suffering from acute inflammation. We demonstrate that excretion of itaconic acid happens at a low rate and that it cannot be detected in significant amounts in plasma or urine of septic patients or in liquid from bronchial lavage of patients with pulmonary inflammation. We conclude that itaconic acid may serve as a pro-inflammatory marker in immune cells but that it does not qualify as a biomarker in the tested body fluids.

Erythritol is a pentose-phosphate pathway metabolite and associated with adiposity gain in young adults.

Metabolomic markers associated with incident central adiposity gain were investigated in young adults. In a 9-mo prospective study of university freshmen (n = 264). Blood samples and anthropometry measurements were collected in the first 3 d on campus and at the end of the year. Plasma from individuals was pooled by phenotype [incident central adiposity, stable adiposity, baseline hemoglobin A1c (HbA1c) > 5.05%, HbA1c < 4.92%] and assayed using GC-MS, chromatograms were analyzed using MetaboliteDetector software, and normalized metabolite levels were compared using Welch's t test. Assays were repeated using freshly prepared pools, and statistically significant metabolites were quantified in a targeted GC-MS approach. Isotope tracer studies were performed to determine if the potential marker was an endogenous human metabolite in men and in whole blood. Participants with incident central adiposity gain had statistically significantly higher blood erythritol [P < 0.001, false discovery rate (FDR) = 0.0435], and the targeted assay revealed 15-fold [95% confidence interval (CI): 13.27, 16.25] higher blood erythritol compared with participants with stable adiposity. Participants with baseline HbA1c > 5.05% had 21-fold (95% CI: 19.84, 21.41) higher blood erythritol compared with participants with lower HbA1c (P < 0.001, FDR = 0.00016). Erythritol was shown to be synthesized endogenously from glucose via the pentose-phosphate pathway (PPP) in stable isotope-assisted ex vivo blood incubation experiments and through in vivo conversion of erythritol to erythronate in stable isotope-assisted dried blood spot experiments. Therefore, endogenous production of erythritol from glucose may contribute to the association between erythritol and obesity observed in young adults.

Body size-dependent Cd accumulation in the zebra mussel Dreissena polymorpha from different routes.

Understanding body size-dependent metal accumulation in aquatic organisms (i.e., metal allometry) is critical in interpreting biomonitoring data. While growth has received the most attention, little is known about controls of metal exposure routes on metal allometry. Here, size-dependent Cd accumulation in zebra mussels (Dreissena polymorpha) from different routes were investigated by exposing mussels to A.(111Cd spiked algae+113Cd spiked river water) or B.(111Cd spiked sediments+113Cd spiked river water). After exposure, 111Cd or 113Cd levels in mussel tissue were found to be negatively correlated with tissue weight, while Cd allometry coefficients (b values) were dependent on Cd exposure routes: -0.664 for algae, -0.241 for sediments and -0.379 for river water, compared to -0.582 in un-exposed mussels. By comparing different Cd exposure routes, we found that size-dependent Cd bioaccumulation from algae or river water could be more responsible for the overall size-dependent Cd accumulation in mussels, and the relative importance of the two sources was dependent on mussel size ranges: Cadmium obtained from algae (algae-Cd) was more important in size-dependent Cd accumulation in smaller mussels (tissue dry weight < 5 mg), while river water-Cd became more important in larger individuals (tissue dry weight > 5 mg). In contrast, sediment-Cd contributed only a small amount to Cd accumulation in zebra mussels and may have little effect on size-dependent Cd bioaccumulation. Our results suggest that size-dependent Cd accumulation in mussels could be largely affected by exposure routes, which should be considered when trying to interpret Cd biomonitoring data of zebra mussels.

Influence of body size on Cu bioaccumulation in zebra mussels Dreissena polymorpha exposed to different sources of particle-associated Cu.

Size of organisms is critical in controlling metal bioavailability and bioaccumulation, while mechanisms of size-related metal bioaccumulation are not fully understood. To investigate the influences of different sources of particle-associated Cu on body size-related Cu bioavailability and bioaccumulation, zebra mussels (Dreissena polymorpha) of different sizes were exposed to stable Cu isotope ((65)Cu) spiked algae (Chlorella vulgaris) or sediments in the laboratory and the Cu tissue concentration-size relationships were compared with that in unexposed mussels. Copper tissue concentrations decreased with mussel size (tissue or shell dry weight) in both unexposed and algal-exposed mussels with similar decreasing patterns, but were independent of size in sediment-exposed mussels. Furthermore, the relative contribution of Cu uptake from algae (65-91%) to Cu bioaccumulation is always higher than that from sediments (9-35%), possibly due to the higher bioavailability of algal-Cu. Therefore, the size-related ingestion of algae could be more important in influencing the size-related variations in Cu bioaccumulation. However, the relative contribution of sediment-Cu to Cu bioaccumulation increased with body size and thus sediment ingestion may also affect the size-related Cu variations in larger mussels (tissue weight >7.5mg). This study highlights the importance of considering exposure pathways in normalization of metal concentration variation when using bivalves as biomonitors.

Temporal and spatial variation in Hg accumulation in zebra mussels (Dreissena polymorpha): possible influences of DOC and diet.

Zebra mussels (Dreissena polymorpha) are filter feeders located near the base of the foodweb and these animals are able to utilize a variety of carbon sources that may also vary seasonally. We conducted both a spatial and a temporal study in order to test the hypotheses: (1) dissolved organic carbon (DOC) concentrations influence Hg accumulation in zebra mussels sampled from a series of lakes and (2) seasonal variations in diet influence Hg accumulation. In the spatial study, we found a significant negative relationship between Hg concentrations and DOC concentrations, suggesting an influence of DOC on Hg bioaccumulation. In the temporal study, we used stable isotope ratios of nitrogen (δ(15)N) and carbon (δ(13)C) as ecological tools to provide a temporally integrated description of the feeding ecology of zebra mussels. Both δ(15)N and δ(13)C varied seasonally in a similar manner: more depleted values occurred in the summer and more enriched values occurred in the fall. Mercury concentrations also varied significantly over the year, with highest concentrations occurring in the summer, followed by a progressive decrease in concentrations into the fall. The C/N ratio of zebra mussels also varied significantly over the year with the lowest values occurring mid-summer and then values increased in the fall and winter, suggesting that there was significant variation in lipid stores. These results indicate that in addition to any effect of seasonal dietary changes, seasonal variation in energy stores also appeared to be related to Hg levels in the zebra mussels. Collectively results from this study suggest that DOC concentrations, seasonal variation in diet and seasonal depletion of energy stores are all important variables to consider when understanding Hg accumulation in zebra mussels.

Influence of contact time and sediment composition on the bioavailability of Cd in sediments.

Stable isotope (111)Cd was spiked into sediments of different organic content levels for 3 days to 2 months. Bioavailability of spiked Cd to deposit-feeders, assessed by in vitro Cd solubilization, generally decreased with contact time but became comparable with that of background Cd after 2 months. This could be explained by the gradual transfer of Cd from the more mobile geochemical phase (carbonate associated phase) to more refractory phases (Fe-Mn oxide associated phase, and organic associated phase) within 2 months. The sedimentary organic content had a weak effect on Cd solubilization, while the distribution of Cd in carbonate or Fe-Mn oxide associated phase could have a larger influence on the solubilization of sedimentary Cd and its change with contact time. The observations in this study emphasize the need to consider Cd sequestration over time in sediments of various compositions, which would be useful in risk assessment of contaminated sediments.

Uranium bioaccumulation in a freshwater ecosystem: impact of feeding ecology.

The objectives of our study were: (1) to determine if there was significant uranium (U) bioaccumulation in a lake that had been historically affected by a U mine and (2) to use a combined approach of gut content examination and stable nitrogen and carbon isotope analysis to determine if U bioaccumulation in fish was linked to foodweb ecology. We collected three species of fish: smallmouth bass (Micropterus dolomieu), yellow perch (Perca flavescens) and bluegill (Lepomis macrochirus), in addition to several invertebrate species including freshwater bivalves (family: Sphaeriidae), dragonfly nymphs (order: Odonata), snails (class: Gastropoda) and zooplankton (family: Daphniidae). Results showed significant U bioaccumulation in the lake impacted by historical mining activities. Uranium accumulation was 2-3 orders of magnitude higher in invertebrates than in the fish species. Within fish, U was measured in operculum (bone), liver and muscle tissue and accumulation followed the order: operculum>liver>muscle. There was a negative relationship between stable nitrogen ratios ((15)N/(14)N) and U bioaccumulation, suggesting U biodilution in the foodweb. Uranium bioaccumulation in all three tissues (bone, liver, muscle) varied among fish species in a consistent manner and followed the order: bluegill>yellow perch>smallmouth bass. Collectively, gut content and stable isotope analysis suggests that invertebrate-consuming fish species (i.e. bluegill) have the highest U levels, while fish species that were mainly piscivores (i.e. smallmouth bass) have the lowest U levels. Our study highlights the importance of understanding the feeding ecology of fish when trying to predict U accumulation.

Effects of aging on the digestive solubilization of Cu from sediments.

Solubilization of particulate Cu by different solutions, mimicking digestive fluids of deposit-feeders, was quantified in stable isotope (65)Cu-spiked sediments (with 3 days-2 months Cu-sediment contact time or aging). Copper solubilization generally decreased with prolonged aging. However, such decrease became less evident after 1 month and equilibrium of Cu in sediments could be reached after 2 months. Aging effects on Cu solubilization can be explained by the changes in Cu geochemical fractionation with aging: Cu generally transferred from more mobile phases (carbonate and Fe-Mn associated) to more refractory phases (organic associated and residual phase). Besides Cu geochemical fractionation, digestive fluid composition and different Cu solubilization pathways involved, as well as sedimentary organic content, could all affect the digestive solubilization of Cu and its change with aging. Our results emphasize the necessity of considering Cu aging in laboratory sediment toxicity experiments, and in risk assessment of Cu contaminated sediments.

The impacts of ontogenetic dietary shifts in yellow perch (Perca flavescens) on Zn and Hg accumulation.

Yellow perch (Perca flavescens) undergo several ontogenetic dietary shifts, and consequently these fish feed at different trophic levels and rely on different carbon sources over their lifetime. Stable isotope ratios of nitrogen (δ(15)N) and carbon (δ(13)C) are powerful ecological tools that are used to provide a temporally integrated description of the feeding ecology of aquatic animals such as fish. The main objective of this study was to use stable isotopes of nitrogen and carbon to determine if dietary changes affected mercury (Hg) and zinc (Zn) accumulation in yellow perch ranging in size from approximately 5 cm to 27 cm. Results showed that Hg bioaccumulation generally increased with increasing trophic level in fish feeding at higher trophic levels, however, the relationship between Hg levels and δ(15)N was non-linear showing no relationship in small fish (less than 15 cm). In contrast, there was a negative, linear relationship between δ(15)N and Zn, suggesting that as perch fed at progressively higher trophic levels, less of Zn bioaccumulated. No relationship was observed between δ(13)C and metal levels in perch. Collectively, these results demonstrate a contrast in the behavior of Zn and Hg bioaccumulation in yellow perch as a function of trophic status.

Seasonal variations in hepatic Cd and Cu concentrations and in the sub-cellular distribution of these metals in juvenile yellow perch (Perca flavescens).

Temporal fluctuations in metal (Cd and Cu) concentrations were monitored over four months (May to August) in the liver of juvenile yellow perch (Perca flavescens) sampled from four lakes situated along a metal concentration gradient in northwestern Quebec: Lake Opasatica (reference lake, low metal concentrations), Lake Vaudray (moderate metal concentrations) and lakes Osisko and Dufault (high metal levels). The objectives of this study were to determine if hepatic metal concentrations and metal-handling strategies at the sub-cellular level varied seasonally. Our results showed that Cd and Cu concentrations varied most, in both absolute and relative values, in fish with the highest hepatic metal concentrations, whereas fish sampled from the reference lake did not show any significant variation. To examine the sub-cellular partitioning of these two metals, we used a differential centrifugation technique that allowed the separation of cellular debris, metal detoxified fractions (heat-stable proteins such as metallothionein) and metal sensitive fractions (heat-denaturable proteins (HDP) and organelles). Whereas Cd concentrations in organelle and HDP fractions were maintained at low concentrations in perch from Lakes Opasatica and Vaudray, concentrations in these sensitive fractions were higher and more variable in perch from Lakes Dufault and Osisko, suggesting that there may be some liver dysfunction in these two fish populations. Similarly, Cu concentrations in these sensitive fractions were higher and more variable in perch from the two most Cu-contaminated lakes (Dufault and Osisko) than in perch from the other two lakes, suggesting a breakdown of homeostatic control over this metal. These results suggest not only that metal concentrations vary seasonally, but also that concentrations vary most in fish from contaminated sites. Furthermore, at the sub-cellular level, homeostatic control of metal concentrations in metal-sensitive fractions is difficult to maintain in perch with high hepatic metal concentrations.

A field study examining metal elimination kinetics in juvenile yellow perch (Perca flavescens).

Currently little is known about how and at what rate fish eliminate metals in natural environments. To address this knowledge gap we examined metal elimination kinetics in the field using juvenile yellow perch (Perca flavescens) that were caught in a metal-contaminated lake (elevated levels of Cd, Cu and Zn) and transplanted to cages held within a reference lake. Fish were sampled from the cages over 75 d and changes in metal concentrations were measured in the gills, gut, liver and kidney. In transplanted fish, Cd concentrations decreased most rapidly in the gills and gut, i.e. from organs in contact with the ambient water and food; biological half lives (t1/2) were 18 and 37 d, respectively, for each organ. Longer half-lives were observed in the liver (75 d) and kidney (52 d) for this metal. Elimination of excess Cu by the liver and gut occurred much more rapidly, with estimated half-lives of labile Cu being 8 and 4 d, respectively, for these two organs. In contrast to Cd and Cu, there was little Zn elimination. To compare how the liver handles different metals during elimination, we used a differential centrifugation approach to examine changes in metal concentrations (Cd and Cu) at the sub-cellular level. Consistent with the long half-life observed for Cd at the whole organ level, there was no significant loss of Cd from any of the sub-cellular fractions. Copper, on the other hand, was lost from both the organelle and cellular debris fractions. As these fractions likely contain structures such as lysosomes, we suggest that Cu is depurated from the liver by direct elimination of these sub-cellular vesicles. These field results clearly demonstrate how the liver handles essential (Cu, Zn) and non-essential metals (Cd) differently during depuration.

Dynamics of Cd, Cu and Zn accumulation in organs and sub-cellular fractions in field transplanted juvenile yellow perch (Perca flavescens).

Juvenile yellow perch (Perca flavescens) were caught in a reference lake and transplanted to cages held within a lake impacted by mining activities, with elevated levels of aqueous bioavailable copper (Cu(2+)), zinc (Zn(2+)) and cadmium (Cd(2+)). Fish were sampled from the cages over 70 d and changes in metal concentrations were followed over time in the gills, gut, liver and kidney. In addition, the hepatic sub-cellular partitioning of the three metals was determined by differential centrifugation of liver samples, yielding the following fractions: cellular debris; organelles; heat-denaturable proteins (HDP); and heat-stable proteins (HSP) (including metallothionein). In transplanted fish, Cd concentrations increased in all the organs sampled, whereas Cu mainly increased in the gills, gut and liver but not the kidney; some slight accumulation of Zn occurred in the kidneys and gills of the transplanted fish. The sub-cellular partitioning results demonstrated that metal-handling strategies in juvenile yellow perch differed among metals. Cellular sequestration in the HSP fraction was an important strategy used by these fish in response to increased ambient Cd. Accumulation of Zn was not seen in the organs examined, indicating that transplanted perch were able to either reduce influx, or increase efflux rates of this metal. The response of yellow perch to elevated ambient Cu appeared to combine the strategies used for Cd and Zn, as both cellular sequestration and reduced accumulation were observed in transplanted fish.

The potential impact of climate on human exposure to contaminants in the Arctic.

Many northern indigenous populations are exposed to elevated concentrations of contaminants through traditional food and many of these contaminants come from regions exterior to the Arctic. Global contaminant pathways include the atmosphere, ocean currents, and river outflow, all of which are affected by climate. In addition to these pathways, precipitation, animal availability, UV radiation, cryosphere degradation and human industrial activities in the North are also affected by climate change. The processes governing contaminant behaviour in both the physical and biological environment are complex and therefore, in order to understand how climate change will affect the exposure of northern people to contaminants, we must have a better understanding of the processes that influence how contaminants behave in the Arctic environment. Furthermore, to predict changes in contaminant levels, we need to first have a good understanding of current contaminant levels in the Arctic environment, biota and human populations. For this reason, it is critical that both spatial and temporal trends in contaminant levels are monitored in the environment, biota and human populations from all the Arctic regions.