Using stable isotope analysis to assess the effects of hypolimnetic oxygenation on diet in a mixed cold- and warmwater fish community

Line-diffuser hypolimnetic oxygenation was initiated in North Twin Lake, Washington, in 2009 to mitigate reductions in Rainbow Trout (Oncorhynchus mykiss) and Brook Trout (Salvelinus fontinalis) habitat due to temperature-dissolved oxygen “habitat squeeze”. Previous studies demonstrated that trout populations rapidly expanded into increased hypolimnetic habitat within the first few years of oxygenation and previous short-term diet analyses indicated an effect on fish diet; however, the long-term effects on fish ecology have yet to be established. In this study, stable isotope analysis of fish liver tissue suggests relatively few differences in feeding ecology of principal coldwater fish species in North Twin Lake compared to unoxygenated South Twin Lake. When compared between lakes, Rainbow Trout and Brook Trout diets contained similar proportions of Daphnia, Chironomidae, and Chaoboridae. Littoral and epilimnetic-focused Golden Shiner (Notemigonus crysoleucas) diets were also similar between lakes. Observed similarities between Golden Shiner and trout diets suggest the effects of interspecific competition between salmonids and non-salmonids may be limiting trout growth and survival. Fisheries managers should therefore consider both habitat limitations and interspecific competition when managing for coldwater fish species in mesotrophic, dimictic lakes.     

Backbone structure of a small helical integral membrane protein: A unique structural characterization

The structural characterization of small integral membrane proteins pose a significant challenge for structural biology because of the multitude of molecular interactions between the protein and its heterogeneous environment. Here, the three‐dimensional backbone structure of Rv1761c from Mycobacterium tuberculosis has been characterized using solution NMR spectroscopy and dodecylphosphocholine (DPC) micelles as a membrane mimetic environment. This 127 residue single transmembrane helix protein has a significant (10 kDa) C‐terminal extramembranous domain. Five hundred and ninety distance, backbone dihedral, and orientational restraints were employed resulting in a 1.16 Å rmsd backbone structure with a transmembrane domain defined at 0.40 Å. The structure determination approach utilized residual dipolar coupling orientation data from partially aligned samples, long‐range paramagnetic relaxation enhancement derived distances, and dihedral restraints from chemical shift indices to determine the global fold. This structural model of Rv1761c displays some influences by the membrane mimetic illustrating that the structure of these membrane proteins is dictated by a combination of the amino acid sequence and the protein's environment. These results demonstrate both the efficacy of the structural approach and the necessity to consider the biophysical properties of membrane mimetics when interpreting structural data of integral membrane proteins and, in particular, small integral membrane proteins.     

Functional expression of NADPH oxidase components (alpha- and beta-subunits of cytochrome b558 and 45-kDa flavoprotein) by intrinsic human glomerular mesangial cells.

Recently, we have shown that human glomerular mesangial cells (HMCs) release oxygen radicals from the plasma membrane in response to cytokines. Now we have used diphenylene iodonium, a covalent binding inhibitor of activated 45-kDa flavoprotein, in neutrophils radiolabeled with 125I and could identify a 45-kDa protein band in a separated HMC plasma membrane fraction. Low temperature difference spectroscopy showed a peak absorbance at 428 and 558 nm. Direct potentiometry of HMC membranes (-340 to -160 mV) showed the presence of a low potential cytochrome (76 pmol/mg to HMC membrane protein) identified as cytochrome b558. In slot blots, mouse monoclonal antibody (mAb) 7D5, specific for the extracellular domain of the alpha-subunit, showed a positive reaction with HMCs. In Western blots, mAb 449, directed against the cytoplasmic epitope of the alpha-subunit, identified a 23-kDa protein; and mAb 48, raised against the large (beta) subunit of cytochrome b558 of human neutrophils (Verhoeven, A. J., Bolscher, B. G. J. M., Meerhof, L. J., van Zwieten, R., Keijer, J., Weening, R. S., and Roos, D. (1989) Blood 73, 1686-1694), detected a smear between 75 and 100 kDa in denatured HMC membrane protein. These data determined with HMCs, suggest for the first time the expression of three essential components of NADPH:O2- oxidoreductase in mesenchymal cells.     

Chemical control of vine weevil larvae on container-grown hardy ornamental nursery stock 1986–1989

In a search for alternatives to the former standard aldrin compost incorporation treatment for control of vine weevil (Otiorhynchus sulcatus) larvae on container-grown hardy ornamental nursery stock, a series of 87 tests of insecticides were done at four experimental centres of the ADAS (Leeds, Reading, Wolverhampton and Wye) from 1986 to 1989. Insecticidally-treated plants and untreated controls were artificially infested with vine weevil eggs at varying intervals before and after treatment, and the survival of the pest was assessed. Aldrin treatment gave consistent and excellent preventive control of vine weevil larvae for over 2 years. Of the candidate materials tested, a slow-release granular formulation of chlorpyrifos incorporated into compost at a dose rate of 100 g a.i. m^-3 of compost gave good control for up to 34 wk after treatment (the longest period evaluated) and a micro-encapsulated slow release formulation of fonofos incorporated at a dose rate of 43.3 g a.i. m^-3 usually gave good control for up to two years (the longest period evaluated). Surface applications of these two organophosphates or of carbofuran granules, though sometimes effective, were unreliable as either preventive or remedial treatments even for short term control.     

In vivo ozone exposure induces antioxidant/stress-related responses in murine lung and skin

Lung and skin are the organs directly exposed to environmental pollution. Ozone (O(3)) is a toxic, oxidant air pollutant, and exposure has been shown to induce antioxidant depletion as well as oxidation of lipids and proteins within the outermost skin layer (stratum corneum) and the lung respiratory tract lining fluids (RTLFs). To further define skin and lung responses to O(3) exposure, SKH-1 hairless mice were exposed to either 0.8 ppm of O(3) (a level occasionally reached in very polluted areas) or ambient air 6 h/day for 6 consecutive days. O(3) exposure resulted in the depletion of alpha-tocopherol in lung and plasma and induction in both skin and lung of heme oxygenase 1, cyclooxygenase 2, and proliferating cell nuclear antigen. O(3)-exposed animals showed a similar extent of upregulation of COX-2 and PCNA in lung and skin, whereas HO-1 was more responsive in skin than in lung (7-fold induction vs. 2-fold induction). In addition to these measures of response to oxidative stress, O(3) exposure led to the activation of nuclear factor kappaB measured as IkappaBalpha phosphorylation in both tissues. We conclude that in this model, O(3) at high pollutant levels is able to affect both lung and skin biology, inducing depletion of alpha-tocopherol and inducing stress-related responses in both skin epidermis and respiratory tract epithelium.     

Consequences of inferring diet from feeding guilds when estimating and interpreting consumer–resource stoichiometry

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Local mutagenesis of Rous sarcoma virus: The major sites of tyrosine and serine phosphorylation of p60src are dispensable for transformation

We have constructed mutants of Rous sarcoma virus expressing p60^src that are underphosphorylated on serine or tyrosine, by linker insertion or insertion/ deletion into cloned Rous sarcoma virus DNA, and recovery of mutant virus by transfection of chicken embryo fibroblasts. Cells infected with mutants whose p60^src lack the major site of either serine or tyrosine phosphorylation were morphologically transformed and formed colonies in soft agar. The tyrosine kinase activities of the mutant p60^src measured in vivo and in vitro were close to the wild type activity. Peptide mapping showed that phosphorylation on tyrosine and serine of p60^src is independent: the major phosphorylated tyrosine and the major phosphorylated serine can each be phosphorylated in the absence of phosphorylation of the other.     

VEGF-receptor signal transduction

The vascular endothelial growth factor (VEGF) family of ligands and receptors has been the focus of attention in vascular biology for more than a decade. There is now a consensus that the VEGFs are crucial for vascular development and neovascularization in physiological and pathological processes in both embryo and adult. This has facilitated a rapid transition to their use in clinical applications, for example, administration of VEGF ligands to enhance vascularization of ischaemic tissues and, conversely, inhibitors of VEGF-receptor function in anti-angiogenic therapy. More recent data indicate essential roles for the VEGFs in haematopoietic cell function and in lymphangiogenesis.     

Effects of Air Pollution and the Introduction of the London Low Emission Zone on the Prevalence of Respiratory and Allergic Symptoms in Schoolchildren in East London: A Sequential Cross-Sectional Study

The adverse effects of traffic-related air pollution on children’s respiratory health have been widely reported, but few studies have evaluated the impact of traffic-control policies designed to reduce urban air pollution. We assessed associations between traffic-related air pollutants and respiratory/allergic symptoms amongst 8–9 year-old schoolchildren living within the London Low Emission Zone (LEZ). Information on respiratory/allergic symptoms was obtained using a parent-completed questionnaire and linked to modelled annual air pollutant concentrations based on the residential address of each child, using a multivariable mixed effects logistic regression analysis. Exposure to traffic-related air pollutants was associated with current rhinitis: NOx (OR 1.01, 95% CI 1.00–1.02), NO₂ (1.03, 1.00–1.06), PM₁₀ (1.16, 1.04–1.28) and PM_2.5 (1.38, 1.08–1.78), all per μg/m³ of pollutant, but not with other respiratory/allergic symptoms. The LEZ did not reduce ambient air pollution levels, or affect the prevalence of respiratory/allergic symptoms over the period studied. These data confirm the previous association between traffic-related air pollutant exposures and symptoms of current rhinitis. Importantly, the London LEZ has not significantly improved air quality within the city, or the respiratory health of the resident population in its first three years of operation. This highlights the need for more robust measures to reduce traffic emissions.