A catchment-based study of endocrine disruption in surface waters: multivariate evaluation of the health of a sentinel fish species exposed to sewage treatment works effluent

Summary of the results in the context of EDCAT 5 project aims: 1. By comparing appropriate biomarkers in fish sampled from STW-impacted sites and control sites during the pre-remediation period, to determine whether there was evidence for any effects that might be attributed to the presence of estrogenic (or androgenic, or anti-androgenic/-estrogenic) endocrine disrupting chemicals in the former. This aim was addressed by measuring concentrations of the estrogen-dependent yolk protein precursor vitellogenin, and the androgen-dependent nest glue spiggin in male and female sticklebacks. In addition histological examination of the gonadal structure of fish captured at the impacted and non-impacted sites was employed to seek evidence of overt alterations in reproductive physiology of the fish. For a subset of matched samples from the two rivers, the relative induction of hepatic choriogenin mRNA, a biomarker of estrogen exposure, was measured. Conclusions: Chemistry data provided by EDCAT3&4 showed that estrogenicity of the effluent was low prior to remediation and lower still following installation of the GAC plant. No evidence of overt estrogenic effects was detected in male sticklebacks in the Ray, VTG and ChG levels were similar in males from both rivers. Nor was there any evidence of alterations in spiggin concentrations in the kidneys of males from the Ray compared to the Ock. However, VTG concentrations in female sticklebacks from the Ray were increased following the STW upgrade as were hepatic ChG transcript levels, and kidney spiggin concentrations. No changes in these elements of the reproductive system were observed in females from the Ock across the same time periods. Chemical analysis of the effluent indicated that prior to installation of the GAC plant substantial concentrations of anti-androgenic chemicals were present, together with a wide range of other organics. Concentrations of these were much reduced following the plant upgrade. It is reasonable to suppose that the changes observed in the female reproductive endocrine system following the upgrade were related to the removal of some or all of this complex mixture of chemicals. The absence of effects in males may be related to the balance between exogenous and endogenous signals, or to the specificity of effects exerted by the chemicals present. No intersex fish were detected from either river. A significant bias in favour of females was detected in the stickleback populations in both rivers suggesting a factor associated with life-history of the fish, rather than contaminant burden, was responsible. 2. By comparing appropriate biomarkers in fish sampled from STW-impacted sites and control sites during the pre-remediation period, to determine whether there was evidence for any effects that might be attributed to the presence of “conventionally” toxic chemicals. This was addressed by measurement of the activity of a key Phase I transforming enzyme in the liver of fish, either using direct enzymatic assay (EROD) or by quantifying the levels of expression of the corresponding gene (CYP1A). Conclusions: EROD activity was significantly greater in fish from the Ray than the Ock in two samples collected prior to the installation of the GAC plant (2006, 2007) and this likely reflects the differential contaminant loading in the two rivers. A single sample following the commissioning of the GAC plant (2008) indicated that EROD activity had increased among fish from the Ock while that in fish from the Ray remained unchanged. While a delayed recovery of this biomarker in fish from the Ray may be expected depending on the route of exposure (direct via water or indirect via contaminated food) the reasons for elevated EROD activity in fish from the Ock/Childrey Brook are not immediately evident. Provision of a full data set for Cyp1A expression awaits the repeat of the assay. When this is complete the factors underlying the EROD findings may become clear. 3. To determine whether the adaptive capacity and energetic status of fish varied between the STW-impacted and non-impacted sites. This was addressed by measurement of indicators of stress (whole-body corticosteroid levels), metabolic status (whole-body glucose levels) and anabolic activity (RNA:DNA ratios). Conclusions: The data provide no evidence that the stress response of fish captured in the Ray prior to installation of the GAC plant was modified by exposure to the effluent. However, large variations in whole-body corticosteroid and glucose concentrations in fish from both rivers, with clear trends over time, were closely linked to perturbations in the river flow regime. Whether there was interaction between environmental and chemical factors in determining corticosteroid and glucose status is difficult to discern but it seems likely that variation in these indicators of the stress axis was driven primarily by environmental factors. The RNA:DNA ratios were closely linked with seasonal change in temperature and closely matched observed patterns of weight and length gain in stickleback populations in the two rivers. The longer growth period enjoyed by fish in the Ray was clearly evident. For both rivers, mean anabolic activity was greater during 2008 than 2007 and it seems likely that this is related to adverse effects associated with the periods of extreme flow change observed on both rivers in 2007. 4. To assess whether there were differences in population size and structure between STW-impacted and non-impacted sites. This was addressed by comparison of key somatic measures, in particular frequency distributions for fork length. Conclusions: Because of the extreme patchiness of the distribution of stickleback populations in both rivers accurate abundance estimates were not obtained. However, the catch per unit effort across the life of the project was similar for both rivers. While population size, and age structure (both rivers hosted annual populations), appeared to be similar fish in the Ray were overall larger than those from the Ock, and spawned earlier. The differences in growth and timing of spawning between the rivers were likely to have been associated with the Rodbourne STW effluent. Downstream of the discharge on the Ray water temperatures were consistently 2 – 3oC above those of the Ock. This temperature difference, in combination with the introduction of additional nutrients into the river which is likely to have affected the availaibility of food, probably accounts for the different growth profile among the sticklebacks in the two rivers. However, over and above this difference, there was a significant increase in size of sticklebacks in the Ray between the matched pre- and post-remediation periods in the Ray while no change in size of the fish in the Ock occurred during the same period. Similarly, the RNA:DNA ratio was higher in fish from the Ock during 2007 but greater in fish from the Ray during 2008. Taken together, these observations suggest that there was an improvement in the status of the fish in the Ray following the commissioning of the GAC plant, while the population in the Ock remained relatively stable. It is reasonable to suppose that this may be linked to the reduction of the chemical load entering the Ray at Rodbourne following the installation of the GAC plant. The Ray is “cleaner” now than was the case prior to remediation but remains nutrient rich and several degrees warmer than the Ock, this combination of factors providing fish in the Ray with greater scope for growth relative to populations in the Ock

Chitobiose utilization in Borrelia burgdorferi is dually regulated by RpoD and RpoS

<p>Abstract</p> <p>Background</p> <p><it>Borrelia burgdorferi </it>has limited biosynthetic capabilities and must scavenge N-acetylglucosamine (GlcNAc), an essential component of the microbial cell wall, from the surrounding environment. Spirochetes cultured in the absence of free GlcNAc exhibit biphasic growth; however, addition of chitobiose (a dimer of GlcNAc) substitutes for free GlcNAc resulting in a single exponential phase. We evaluated the effect of RpoS and RpoN, the only alternative sigma factors in <it>B. burgdorferi</it>, on biphasic growth and chitobiose utilization in the absence of free GlcNAc. In addition, we investigated the source of GlcNAc in the second exponential phase.</p> <p>Results</p> <p>By comparing the growth of wild-type cells to insertional mutants for <it>rpoS </it>and <it>rpoN </it>we determined that RpoS, but not RpoN, partially regulates both biphasic growth and chitobiose utilization. The <it>rpoS </it>mutant, cultured in the absence of free GlcNAc, exhibited a significant delay in the ability to initiate a second exponential phase compared to the wild type and <it>rpoS </it>complemented mutant. Expression analysis of <it>chbC</it>, which encodes the membrane-spanning protein of the chitobiose phosphotransferase system, suggests the delay is due to the inability of the <it>rpoS </it>mutant to up regulate <it>chbC</it>. Furthermore, supplementing GlcNAc starved cultures with high concentrations (75 or 150 μM) of chitobiose resulted in biphasic growth of the <it>rpoS </it>mutant compared to a single exponential phase for the wild type and <it>rpoS </it>complemented mutant. In contrast, growth of the <it>rpoN </it>mutant under all conditions was similar to the wild type. 5' Rapid amplification of cDNA ends (5' RACE) revealed the transcriptional start site for <it>chbC </it>to be 42 bp upstream of the translational start site. Analysis of the <it>chbC </it>promoter region revealed homology to previously described RpoD and RpoS <it>B. burgdorferi </it>promoters. We also determined that yeastolate, a component of the growth medium (BSK-II), is not essential for second exponential phase growth.</p> <p>Conclusion</p> <p>Together these results suggest that RpoD and RpoS, but not RpoN, regulate biphasic growth and chitobiose utilization in <it>B. burgdorferi </it>by regulating the expression of the chitobiose transporter (<it>chbC</it>). The data also demonstrate that the second exponential phase observed in wild-type cells in the absence of free GlcNAc is not due to free chitobiose or GlcNAc oligomers present in the medium.</p

The chitobiose transporter, chbC, is required for chitin utilization in Borrelia burgdorferi

<p>Abstract</p> <p>Background</p> <p>The bacterium <it>Borrelia burgdorferi</it>, the causative agent of Lyme disease, is a limited-genome organism that must obtain many of its biochemical building blocks, including N-acetylglucosamine (GlcNAc), from its tick or vertebrate host. GlcNAc can be imported into the cell as a monomer or dimer (chitobiose), and the annotation for several <it>B. burgdorferi </it>genes suggests that this organism may be able to degrade and utilize chitin, a polymer of GlcNAc. We investigated the ability of <it>B. burgdorferi </it>to utilize chitin in the absence of free GlcNAc, and we attempted to identify genes involved in the process. We also examined the role of RpoS, one of two alternative sigma factors present in <it>B. burgdorferi</it>, in the regulation of chitin utilization.</p> <p>Results</p> <p>Using fluorescent chitinase substrates, we demonstrated an inherent chitinase activity in rabbit serum, a component of the <it>B. burgdorferi </it>growth medium (BSK-II). After inactivating this activity by boiling, we showed that wild-type cells can utilize chitotriose, chitohexose or coarse chitin flakes in the presence of boiled serum and in the absence of free GlcNAc. Further, we replaced the serum component of BSK-II with a lipid extract and still observed growth on chitin substrates without free GlcNAc. In an attempt to knockout <it>B. burgdorferi </it>chitinase activity, we generated mutations in two genes (<it>bb0002 </it>and <it>bb0620</it>) predicted to encode enzymes that could potentially cleave the β-(1,4)-glycosidic linkages found in chitin. While these mutations had no effect on the ability to utilize chitin, a mutation in the gene encoding the chitobiose transporter (<it>bbb04</it>, <it>chbC</it>) did block utilization of chitin substrates by <it>B. burgdorferi</it>. Finally, we provide evidence that chitin utilization in an <it>rpoS </it>mutant is delayed compared to wild-type cells, indicating that RpoS may be involved in the regulation of chitin degradation by this organism.</p> <p>Conclusions</p> <p>The data collected in this study demonstrate that <it>B. burgdorferi </it>can utilize chitin as a source of GlcNAc in the absence of free GlcNAc, and suggest that chitin is cleaved into dimers before being imported across the cytoplasmic membrane via the chitobiose transporter. In addition, our data suggest that the enzyme(s) involved in chitin degradation are at least partially regulated by the alternative sigma factor RpoS.</p

Gaussian multiplicative Chaos for symmetric isotropic matrices

Motivated by isotropic fully developed turbulence, we define a theory of symmetric matrix valued isotropic Gaussian multiplicative chaos. Our construction extends the scalar theory developed by J.P. Kahane in 1985

Aluminum Abundance on the Surface of Mercury: Application of a New Background-Reduction Technique for the Analysis of Gamma-Ray Spectroscopy Data

A new technique has been developed for characterizing gamma-ray emission from a planetary surface in the presence of large background signals generated in a spacecraft. This technique is applied to the analysis of Al gamma rays measured by the MESSENGER Gamma-Ray Spectrometer to determine the abundance of Al on the surface of Mercury. The result (Al/Si = 0.29-0.13+0.05) is consistent with Al/Si ratios derived from the MESSENGER X-Ray Spectrometer and confirms the finding of low Al abundances. The measured abundance rules out a global, lunar-like feldspar-rich crust and is consistent with previously suggested analogs for surface material on Mercury, including terrestrial komatiites, low-iron basalts, partial melts of CB chondrites, and partial melts of enstatite chondrites. Additional applications of this technique include the measurement of other elements on Mercury's surface as well as the analysis of data from other planetary gamma-ray spectrometer experiments

Association Between Objectively Measured Physical Activity and Cognitive Function in Older Adults—The Reasons for Geographic and Racial Differences in Stroke Study

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/116310/1/jgs13829_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/116310/2/jgs13829.pd