Reactivity and survivability of glycolaldehyde in simulated meteorite impact experiments.

Sugars of extraterrestrial origin have been observed in the interstellar medium (ISM), in at least one comet spectrum, and in several carbonaceous chondritic meteorites that have been recovered from the surface of the Earth. The origins of these sugars within the meteorites have been debated. To explore the possibility that sugars could be generated during shock events, this paper reports on the results of the first laboratory impact experiments wherein glycolaldehyde, found in the ISM, as well as glycolaldehyde mixed with montmorillonite clay, have been subjected to reverberated shocks from ~5 to >25 GPa. New biologically relevant molecules, including threose, erythrose and ethylene glycol, were identified in the resulting samples. These results show that sugar molecules can not only survive but also become more complex during impact delivery to planetary bodies.

Mitigation of two pyrethroid insecticides in a Mississippi Delta constructed wetland.

Constructed wetlands are a suggested best management practice to help mitigate agricultural runoff before entering receiving aquatic ecosystems. A constructed wetland system (180 m x 30 m), comprising a sediment retention basin and two treatment cells, was used to determine the fate and transport of simulated runoff containing the pyrethroid insecticides lambda-cyhalothrin and cyfluthrin, as well as suspended sediment. Wetland water, sediment, and plant samples were collected spatially and temporally over 55 d. Results showed 49 and 76% of the study's measured lambda-cyhalothrin and cyfluthrin masses were associated with vegetation, respectively. Based on conservative effects concentrations for invertebrates and regression analyses of maximum observed wetland aqueous concentrations, a wetland length of 215 m x 30 m width would be required to adequately mitigate 1% pesticide runoff from a 14 ha contributing area. Results of this experiment can be used to model future design specifications for constructed wetland mitigation of pyrethroid insecticides.

Mitigation of azinphos-methyl in a vegetated stream: comparison of runoff- and spray-drift.

The effectiveness of aquatic macrophytes in reducing runoff- and spray-drift-induced azinphos-methyl (AZP) input was compared in a vegetated stream. Water, sediment and plant samples were taken at increasing distances from a point of input during a spray-drift event and two runoff (10 and 22 mm/day) events. Peak concentrations of AZP decreased significantly (R2=0.99; p<0.0001; n=5) from 0.24 microg/l to 0.11 microg/l during the 10mm runoff event. No reduction took place during the 22 mm event. AZP concentrations were reduced by 90% following spray-drift input, with peak concentrations decreasing significantly (R2=0.93; p=0.0084; n=5) from 4.3 microg/l to 1.7 microg/l with increasing distance from the point of input. Plant samples taken after the spray-drift event showed increased AZP concentrations in comparison to before the event indicating sorption of the pesticide to the macrophytes. Although peak concentrations of AZP were as effectively mitigated during the 10mm runoff event as during the spray-drift event, predictive modelling revealed that maximum concentrations expected during a worst-case scenario 10mm runoff event (0 days after application) are an order of magnitude lower than what can be expected for a worst-case spray-drift and 22 mm runoff event, suggesting that spray-drift-derived pesticide concentrations are more effectively mitigated than those of runoff.

Influence of vegetation in mitigation of methyl parathion runoff.

A pesticide runoff event was simulated on two 10 m x 50 m constructed wetlands (one non-vegetated, one vegetated) to evaluate the fate of methyl parathion (MeP) (Penncap-M). Water, sediment, and plant samples were collected at five sites downstream of the inflow for 120 d. Semi-permeable membrane devices (SPMDs) were deployed at each wetland outflow to determine exiting pesticide load. MeP was detected in water at all locations of the non-vegetated wetland (50 m), 30 min post-exposure. MeP was detected 20 m from the vegetated wetland inflow 30 min post-exposure, while after 10d it was detected only at 10 m. MeP was measured only in SPMDs deployed in non-vegetated wetland cells, suggesting detectable levels were not present near the vegetated wetland outflow. Furthermore, mass balance calculations indicated vegetated wetlands were more effective in reducing aqueous loadings of MeP introduced into the wetland systems. This demonstrates the importance of vegetation as sorption sites for pesticides in constructed wetlands.

Defining the educational needs of recent dental graduates preparing for the Membership of the Faculty of Dental Surgery examination.

OBJECTIVES: On graduation, UK dentists wishing to advance their career enter two years of general professional training aimed at consolidating their undergraduate experience. The Membership of the Faculty of Dental Surgery examination (MFDS) attests to its successful completion and is a pre-requisite for entry into training programmes which lead to specialist status. Most MFDS candidates prepare for the examination on their own while in full-time employment and many reinforce this self-directed learning with participation in short revision courses or through distance learning. Here we seek to obtain data on the specific educational needs of these individuals. METHODS: Questionnaires were used to interrogate 92 UK graduates attending short MFDS revision courses of up to 1 week's duration to identify which topic areas were perceived as particular areas of weakness. To gain greater insight into the responses obtained, 18/92 respondents were selected at random and followed up with semi-structured interviews informed by the questionnaires. RESULTS: Basic medical science, human diseases, law and ethics and health and safety regulations were the areas of weakness most frequently highlighted by the respondents. Most had undergone comprehensive courses in the first two topics; however, the interviews suggested that this was generally in the early stages of undergraduate training when they had difficulty in contextualising large quantities of new information. In the case of the latter two, teaching had been very varied and several interviewees felt that it had been inadequate. CONCLUSION: Recent graduates preparing for MFDS have clear educational needs. These data have begun to characterise the requirements of this group and may inform the planning of short revision courses designed to assist them.

Evaluating acute toxicity of methyl parathion application in constructed wetland mesocosms.

Wetland ecosystems have reduced ambient levels of various organic and metallic compounds, although their effectiveness on agricultural pesticides is not well documented. Five stations within each of two 10 x 50 m constructed wetlands (two vegetated, two nonvegetated) were selected to measure the fate and effects of methyl parathion (MeP). Following a simulated storm event (0.64 cm of rainfall), aqueous, sediment, and plant samples were collected and analyzed spatially (5, 10, 20, and 40 m from the inlet) and temporally (after 3-10 days) for MeP concentrations and for the impact of those concentrations on the aquatic fauna. Aqueous toxicity to fish decreased spatially and temporally in the vegetated mesocosm. Pimephales promelas survival was significantly reduced, to 68%, at the 10-m station of the nonvegetated wetlands (3 h postapplication), with pesticide concentrations averaging 9.6 microg MeP/L. Ceriodaphnia in both the vegetated and nonvegetated wetlands was sensitive (i.e., a significant acute response to MeP occurred) to pesticide concentrations through 10 days postapplication. Mean MeP concentrations in water ranged from 0.5 to 15.4 microg/L and from 0.1 to 27.0 microg/L in the vegetated and nonvegetated wetlands, respectively. Hyalella azteca aqueous tests resulted in significant mortality in the 5-m vegetated segment 10 days after exposure to MeP (2.2 microg/L). Solid-phase (10-day) sediment toxicity tests showed no significant reduction in Chironomus tentans survival or growth, except for the sediments sampled 3 h postapplication in the nonvegetated wetland (65% survival). Thereafter, midge survival averaged >87% in sediments sampled from both wetlands. These data suggest that wetlands play a significant role in mitigating the effect of MeP exposure in sensitive aquatic biota.

Hydroxylated and methyl sulfone PCB metabolites in adipose and whole blood of polar bear (Ursus maritimus) from East Greenland.

Persistent methyl sulfone (MeSO2-) and hydroxylated (HO-) polychlorinated biphenyl (PCB) metabolites have emerged as important classes of environmental contaminants in vertebrate, aquatic biota and humans. In the present study, PCB, MeSO2-PCB and HO-PCB concentrations and congener patterns were determined in the whole blood and adipose tissue of male (n = 7) and female (n = 12) polar bears (Ursus maritimus) of random age (3-25 years of age), and collected in 1999-2001 from the Ittoqqortoormiit/Scoresby Sound area in central East Greenland. There was no significant difference (P < 0.05) between males and females with respect to PCB or PCB metabolite concentrations in either tissue. The mean sum (Sigma) PCB concentrations were 7020+/-3366 ng/g lipid weight (lw) (range 2708-18148 ng/g lw) and 46.1+/-44.6 ng/g wet weight (ww) (range 12.6-204.2 ng/g ww) in adipose and blood, respectively. The mean Sigma-HO-PCB concentration in whole blood was 182.3+/-72.1 ng/g ww (range 93.8-382.1 ng/g ww). The mean Sigma-HO-PCB to Sigma-PCB concentration ratios in whole blood were 4.59+/-3.58 (range 1.03-11.88) and 8.30+/-5.56 (range 2.16-19.47) in females and males, respectively, which are the highest ratios reported so far for polar bears from any population, or for any free-ranging animal. Sigma-HO-PCB concentrations were greater than all other major classes of organochlorines (i.e. Sigma-PCBs, Sigma-MeSO2-PCBs, Sigma-chlordanes (CHLs), Sigma-hexachlorocyclohexanes (HCHs) and Sigma-chlorobenzenes (CBzs). The mean Sigma-MeSO2-PCB concentrations were 699+/-836 ng/g lw (range 127-3920 ng/g lw) and 10.9+/-9.6 ng/g ww (range 4.3-52.1 ng/g ww) in the adipose and blood, respectively. Regardless of age and sex, in both adipose and whole blood the MeSO2-PCB congener pattern was dominated by 3'- and 4'-MeSO2-CB101 and -CB87, and 4-MeSO2-CB149 (approx. 70% of the Sigma-MeSO2-PCBs). Minor differences in the MeSO2-PCB congener pattern were observed between blood and adipose, which suggests the possible influence of metabolite structure on mobilization and/or deposition to the adipose tissue. Sixteen HO-PCB congeners and one di-HO-PCB congener were identified, and five HO-PCB isomers and one di-HO-PCB isomer were detected. However, congener patterns were dominated by 4'-OH-CB120, 4-HO-CB146/3-HO-CB153, 4-OH-CB187, 4'-HO-CB172, 4-HO-CB193 and 4,4'-di-HO-CB202 (> 10 ng/g ww). HO-PCB congener patterns in whole blood were not significantly different (P < 0.05) between males and females. Other chlorinated phenolic contaminants, pentachlorophenol (0.3+/-0.3 ng/g ww) and 4-HO-heptachlorostyrene (7.5+/-2.9 ng/g ww) were also detected in blood. To our knowledge, this is to first report comparing PCBs, MeSO2-PCBs and HO-PCBs in whole blood and adipose tissue in a free-ranging wildlife species. HO-PCBs and MeSO2-PCBs are both important circulating contaminants in polar bears from this eastern Greenland population. Given the known toxicities of PCB metabolites, this population of polar bear may be experiencing health risks due to exposure to a complex loading of organohalogen contaminants that include HO-PCB and MeSO2-PCB metabolites.

Innovative uses of vegetated drainage ditches for reducing agricultural runoff.

Vegetated agricultural ditches play an important role in mitigation of pesticides following irrigation and storm runoff events. In a simulated runoff event in the Mississippi (USA) Delta, the mitigation capacity of a drainage ditch using the pyrethroid esfenvalerate (Asana XL) was evaluated. The pesticide was amended to soil prior to the runoff event to simulate actual runoff, ensuring the presence of esfenvalerate in both water and suspended particulate phases. Water, sediment, and plant samples were collected temporally and spatially along the drainage ditch. Even with mixing of the pesticide with soil before application, approximately 99% of measured esfenvalerate was associated with ditch vegetation (Ludwigia peploides, Polygonum amphibium, and Leersia oryzoides) three hours following event initiation. This trend continued for the 112 d study duration. Simple modeling results also suggest that aqueous concentrations of esfenvalerate could be mitigated to 0.1% of the initial exposure concentration within 510 m of a vegetated ditch. Observed field half-lives in water, sediment, and plant were 0.12 d, 9 d, and 1.3 d, respectively. These results validate the role vegetation plays in the mitigation of pesticides, and that ditches are an indispensable component of the agricultural production landscape.

Acute toxicity of methyl-parathion in wetland mesocosms: assessing the influence of aquatic plants using laboratory testing with Hyalella azteca.

Methyl-parathion (MeP) was introduced into constructed wetlands for the purpose of assessing the importance of distance from the source of contamination and the role of emergent vegetation on the acute toxicity to Hyalella azteca (Crustacea: Amphipoda). A vegetated (90% cover: mainly Juncus effuses) and a nonvegetated wetland (each with a water body of 50 x 5.5 x 0.2 m) were each exposed to a simulated MeP storm runoff event. H. azteca was exposed for 48 h in the laboratory to water samples taken from the wetlands at a distance of 5, 10, 20, and 40 m from the pesticide inlet 3 h, 24 h, 96 h, and 10 days following application. Methyl-parathion was detected throughout the nonvegetated wetland, whereas the pesticide was only transported halfway through the vegetated wetland. A repeated-measure three-way analysis of variance (ANOVA) using time, location, and vegetation indicated significantly lower toxicity in the vegetated wetland. Furthermore, the mortality decreased significantly with both increasing distance from the inlet and time (48-h LC50 +/- 95% CI: 9.0 +/- 0.3 microg/L). A significant three-way interaction of time x vegetation x location confirmed higher toxicity at the inlet area of the nonvegetated wetland immediately after contamination. Significant linear regressions of maximum mortality (independent of time) versus distance from the pesticide inlet indicated that 44 m of vegetated and 111 m of nonvegetated wetland would reduce H. azteca mortality to < or = 5%. These results suggest that vegetation contributes to reduced MeP effects in constructed wetlands.

Protective effect of apolipoprotein E-mimetic peptides on N-methyl-D-aspartate excitotoxicity in primary rat neuronal-glial cell cultures.

Apolipoprotein E (apoE) is a 34-kD protein with multiple biological properties. Recent clinical and preclinical observations implicate a role for apoE in modifying the response of the brain to focal and global ischemia. One mechanism by which apoE might exert these effects is by reducing glutamate-induced excitotoxic neuronal injury associated with ischemic insults. We demonstrate that human recombinant apoE confers a mild neuroprotective effect in primary neuronal-glial cultures exposed to 100 microM N-methyl-D-aspartate. Furthermore, a peptide derived from the receptor-binding region of apoE (residues 133-149) maintained a significant helical population as assessed by circular dichroism, and completely suppressed the neuronal cell death and calcium influx associated with N-methyl-D-aspartate exposure. Neuroprotection was greatest when the peptide was added concurrently with N-methyl-D-aspartate; however, a significant protection was observed when peptide was preincubated and washed off prior to N-methyl-D-aspartate exposure. These results suggest that one mechanism by which apoE may modify the CNS response to ischemia is by partially blocking glutamate excitotoxicity. Moreover, small peptide fragments derived from the receptor-binding region of apoE have enhanced bioactivity compared with the intact holoprotein, and may represent a novel therapeutic strategy for the treatment of brain ischemia.

Downregulation of microglial activation by apolipoprotein E and apoE-mimetic peptides.

Apolipoprotein E plays an important role in recovery from acute brain injury and risk of developing Alzheimer's disease. We demonstrate that biologically relevant concentrations of apoE suppress microglial activation and release of TNFalpha and NO in a dose-dependent fashion. Peptides derived from the apoE receptor-binding region mimic the effects of the intact protein, whereas deletion of apoE residues 146-149 abolishes peptide bioactivity. These results are consistent with the hypothesis that apoE modulates microglial function by binding specific cell surface receptors and that the immunomodulatory effects of apoE in the central nervous system may account for its role in acute and chronic neurological disease.

Mutation of arginine 44 of GAT-1, a (Na(+) + Cl(-))-coupled gamma-aminobutyric acid transporter from rat brain, impairs net flux but not exchange.

The gamma-aminobutyric acid (GABA) transporter GAT-1 is a prototype of a large family of neurotransmitter transporters that includes those of dopamine and serotonin. GAT-1 maintains low synaptic concentrations of neurotransmitter by coupling GABA uptake to the fluxes of sodium and chloride. Here we identify a stretch of four amino acid residues predicted to lie in the juxtamembrane region prior to transmembrane domain 1 in the cytoplasmic amino-terminal tail of GAT-1, which is critical for its function. Two residues, arginine 44 and tryptophan 47, are fully conserved within the transporter family, and their deletion abolishes GABA transport in the HeLa cell expression system used. Tryptophan 47 can be replaced only by aromatic residues without loss of activity. Arginine 44 is essential for activity. Only when it is replaced by lysine, low activity levels (around 15% of those of the wild type) are observed. Using a reconstitution assay, we show that mutants in which this residue is replaced by lysine or histidine exhibit sodium- and chloride-dependent GABA exchange similar to the wild type. This indicates that these mutants are selectively impaired in the reorientation of the unloaded transporter, a step in the translocation cycle by which net flux and exchange differ. The high degree of conservation in the consensus sequence RXXW suggests that this region may influence the reorientation step in related transporters as well.

Exposure to environmentally relevant concentrations of different nonylphenol formulations in Japanese medaka.

The time course of exposure to p-nonylphenol (NP) from two different sources was compared to equivalent exposures of 17-beta-estradiol (E2) and a solvent control (ethanol; EtOH). Japanese medaka were exposed for 4 days to a nominal concentration of 20 micrograms/l of either NP-I (Schenectady International, Inc.), NP-II (Aldrich), or E2, and were then placed in untreated water for 5 days. Tissue samples were taken at two time points during the 4-day exposure and two time points during the 5 days following exposure. Liver homogenates were analyzed using a western blot to detect vitellogenin (VTG) and quantified by measuring the optical density for each lane. Preliminary results indicate that E2 significantly increased VTG staining above the level observed in EtOH-treated controls for both males and females. A two-way analysis of variance (ANOVA) indicates that NP from both sources, as well as E2, significantly increased VTG staining in males (ANOVA, n = 48, P < 0.001; Tukey pairwise tests, all P < 0.008). A significant increase in VTG was observed in E2-treated males and females the first day following transfer into toxicant-free water (two-way ANOVAs, both n = 48, P < 0.003; Tukey pairwise tests, all P < 0.019). If confirmed, this extended response observed for low-level exposures may represent a significant factor for sampling scenarios following pulsitile exposure.

Developmental evaluation of a potential non-steroidal estrogen: triclosan.

Triclosan is an antibacterial agent commonly used in industry and often detected in waste-water effluent. The potential of triclosan to act as an endocrine disruptor was examined because its chemical structure closely resembles known non-steroidal estrogens (e.g. DES, bisphenol A). Japanese medaka fry (Oryzias latipes) were exposed for 14 days beginning 2 days post-hatch to triclosan (100, 10, 1 micrograms/l), 17-beta estradiol (E2; 1 microgram/l), or a solvent control (ethanol). Two months post-exposure, the phenotypic sex of each adult was assessed visually using sexually dimorphic fin shape and size. The proportion of females in each group was similar for triclosan-exposed animals and solvent-treated controls (ethanol 53%, 1 ppb 58%, 10 ppb 45%, 100 ppb 36%) although E2 treatment did produce 92% female adults. Sexually dimorphic fin traits were quantified to look for potential effects of triclosan and E2 on the development of secondary sexual characters. These results do not support the hypothesis that triclosan is potently estrogenic. However, changes in fin length and non-significant trends in sex ratio suggest triclosan is potentially weakly androgenic.

Endogenous apolipoprotein E suppresses LPS-stimulated microglial nitric oxide production.

The human apolipoprotein (apo) E4 isoform is associated with an increased risk for Alzheimer's disease (AD) and poor prognosis after acute CNS injury. Addition of human apoE inhibits murine microglial activation in culture, suggesting that microglia might be an important physiological target of apoE. In the present study, we examined the role of endogenous murine apoE in modulating microglial nitric oxide (NO) production following lipopolysaccharide (LPS) stimulation. Brain cultures from apoE-deficient mouse pups showed enhanced NO production relative to cultures from wild-type mice and from transgenic mice expressing the human apoE3 isoform, demonstrating that endogenous apoE produced by glial cultures is capable of inhibiting microglial function. ApoE produced within the brain may suppress microglial reactivity and thus alter the CNS response to acute and chronic injury.

Apolipoprotein E suppresses glial cell secretion of TNF alpha.

Apolipoprotein E (apoE) is a 299 amino acid protein with multiple biological functions. Initially described in the context of cholesterol metabolism, apoE also has immunomodulatory properties and recent evidence has implicated a role for apoE in neurological disease. One possibility is that apoE, which is the predominant apolipoprotein produced intra-axially, may modify the CNS response to acute and chronic injury. We prepared mixed neuronal-glial cultures from apoE deficient mouse pups and measured secretion of TNF alpha after stimulation with lipopolysaccharide (LPS) in the presence and absence of human recombinant apoE3 and E4. We demonstrate that preincubation with apoE blocks glial secretion of TNF alpha in a dose-dependent manner. This effect is independent of any direct effect of apoE on cell viability and is greatest when apoE is preincubated with the cell culture for 24 h.

The membrane topology of GAT-1, a (Na+ + Cl-)-coupled gamma-aminobutyric acid transporter from rat brain.

The membrane topology of GAT-1, a sodium- and chloride-coupled gamma-aminobutyric acid transporter from rat brain, has been probed using N-glycosylation scanning mutagenesis. Overall, the results support the theoretical 12-transmembrane segment model. This model (based on hydropathy analysis) was originally proposed for GAT-1 and adopted for all other members of the sodium- and chloride-dependent neurotransmitter transporter superfamily. However, our data indicate that the loop connecting putative transmembrane domains 2 and 3, which was predicted to be located intracellularly, can be glycosylated in vivo. Furthermore, studies with permeant and impermeant methanesulfonate reagents suggest that cysteine 74, located in the hydrophilic loop connecting transmembrane domains 1 and 2, is intracellular rather than extracellular. We present a model in which the topology deviates from the theoretical one in the amino-terminal third of the transporter. It also contains 12 transmembrane segments, but the highly conserved domain 1 does not form a conventional transmembrane alpha-helix.