Response of the benthic fauna of an urban stream during six years of restoration

Okeover Stream flows through the University of Canterbury campus and has been subject to restoration since 1998. While initially spring-fed, its main source of flow is now aquifer water, which has been used for cooling university buildings. Water quality is generally good, but the low-gradient streambed includes substantial amounts of fine inorganic sediment and organic matter including deciduous tree leaves. Restoration activities include riparian plantings, channel shaping, substratum manipulations and additions, the construction of sediment traps and macrophyte management. Thirty aquatic invertebrate taxa (13-19 per year) have been recorded in annual surveys since 2000. Paracalliope fluviatilis (Amphipoda), Copepoda and Oligochaeta were most abundant in all years, whereas Mollusca and Trichoptera always made up <4 % and <2 % of individuals, respectively. Furthermore, cased caddisflies were found only in the two (of four) downstream reaches, whereas Copepoda were predominantly in the upper two reaches where flow was generally slower. Low annual MCI (69-84) and SQMCI (3.5-4.8) values indicated the fauna comprised mainly species that are tolerant of poor water quality or degraded habitat conditions. Our data indicate that the invertebrate fauna has yet to respond positively to the changes in physical habitat and riparian conditions made along Okeover Stream. The introduction of pulses of poor quality water during heavy rainfalls, high levels of siltation, heavy metals in bed sediments, large accumulations of slowly decomposing leaves and an inadequate source of potential colonists may all contribute to the weak response of the invertebrate fauna to restoration activities

Peroxidasin protein expression and enzymatic activity in metastatic melanoma cell lines are associated with invasive potential

Peroxidasin, a heme peroxidase, has been shown to play a role in cancer progression. mRNA expression has been reported to be upregulated in metastatic melanoma cell lines and connected to the invasive phenotype, but little is known about how peroxidasin acts in cancer cells. We have analyzed peroxidasin protein expression and activity in eight metastatic melanoma cell lines using an ELISA developed with an in-house peroxidasin binding protein. RNAseq data analysis confirmed high peroxidasin mRNA expression in the five cell lines classified as invasive and low expression in the three non-invasive cell lines. Protein levels of peroxidasin were higher in the cell lines with an invasive phenotype. Active peroxidasin was secreted to the cell culture medium, where it accumulated over time, and peroxidasin protein levels in the medium were also much higher in invasive than non-invasive cell lines. The only well-established physiological role of peroxidasin is in the formation of a sulfilimine bond, which cross-links collagen IV in basement membranes via catalyzed oxidation of bromide to hypobromous acid. We found that peroxidasin secreted from melanoma cells formed sulfilimine bonds in uncross-linked collagen IV, confirming peroxidasin activity and hypobromous acid formation. Moreover, 3-bromotyrosine, a stable product of hypobromous acid reacting with tyrosine residues, was detected in invasive melanoma cells, substantiating that their expression of peroxidasin generates hypobromous acid, and showing that it does not exclusively react with collagen IV, but also with other biomolecules

The myeloperoxidase-derived oxidant HOSCN inhibits protein tyrosine phosphatases and modulates cell signalling via the mitogen-activated protein kinase (MAPK) pathway in macrophages

MPO (myeloperoxidase) catalyses the oxidation of chloride, bromide and thiocyanate by hydrogen peroxide to HOCl (hypochlorous acid), HOBr (hypobromous acid) and HOSCN (hypothiocyanous acid) respectively. Specificity constants indicate that SCN− is a major substrate for MPO. HOSCN is also a major oxidant generated by other peroxidases including salivary, gastric and eosinophil peroxidases. While HOCl and HOBr are powerful oxidizing agents, HOSCN is a less reactive, but more specific, oxidant which targets thiols and especially low pKa species. In the present study we show that HOSCN targets cysteine residues present in PTPs (protein tyrosine phosphatases) with this resulting in a loss of PTP activity for the isolated enzyme, in cell lysates and intact J774A.1 macrophage-like cells. Inhibition also occurs with MPO-generated HOCl and HOBr, but is more marked with MPO-generated HOSCN, particularly at longer incubation times. This inhibition is reversed by dithiothreitol, particularly at early time points, consistent with the reversible oxidation of the active site cysteine residue to give either a cysteine–SCN adduct or a sulfenic acid. Inhibition of PTP activity is associated with increased phosphorylation of p38a and ERK2 (extracellular-signal-regulated kinase 2) as detected by Western blot analysis and phosphoprotein arrays, and results in altered MAPK (mitogen-activated protein kinase) signalling. These data indicate that the highly selective targeting of some protein thiols by HOSCN can result in perturbation of cellular phosphorylation and altered cell signalling. These changes occur with (patho)physiological concentrations of SCN− ions, and implicate HOSCN as an important mediator of inflammation-induced oxidative damage, particularly in smokers who have elevated plasma levels of SCN−

Mayfly production in a New Zealand glacial stream and the potential effect of climate change

Abstract In contrast to the northern hemisphere where species of Chironomidae are usually the dominant benthic invertebrates in the coldest upper reaches of glacial streams, mayflies (Deleatidium spp.: Leptophlebiidae) predominate in equivalent conditions in New Zealand. We examined the life histories and annual production of Deleatidium spp. at two sites on the Matukituki River (South Island, New Zealand) and at three sites in its glacier-fed tributary, Rob Roy Stream. Mean annual water temperature at the five sites ranged from 2.1 to 7.0°C. Monthly sampling showed that mayfly populations were poorly synchronised at all sites but were probably univoltine. The large Deleatidium cornutum was the dominant mayfly species found at the upper sites (Sites 1 and 2) on Rob Roy Stream, whereas above the confluence with Matukituki River (Site 3) it co-existed with a complex of smaller species we refer to as D. ''angustum''. Deleatidium ''angustum'' also dominated at the Matukituki sites. Deleatidium production calculated for the five sites, assuming an 11-month nymphal life, ranged from 0.48 g dry weight/m 2 /year (Site 1) to 3.07 g dry weight/m 2 /year (Site 3). The values for D. cornutum at Sites 2 and 3 are high for a species of Deleatidium and reflect its large size. This species appears to be strongly adapted for growth at low temperatures. Climate change scenarios for New Zealand predict the gradual and ultimate loss of small South Island glaciers and a consequent warming of streams as runoff from rainfall and snow melt becomes more dominant in spring. As a result, suitable habitats will be lost for cold-water specialists such as D. cornutum, and they are likely to suffer reductions in their distributional range and local extinction. In contrast, species such as those in the D. ''angustum'' complex may extend their ranges into streams formerly dominated by glacial meltwater

Selenium-containing amino acids are targets for myeloperoxidase-derived hypothiocyanous acid: determination of absolute rate constants and implications for biological damage

Elevated MPO (myeloperoxidase) levels are associated with multiple human inflammatory pathologies. MPO catalyses the oxidation of Cl−, Br− and SCN− by H2O2 to generate the powerful oxidants hypochlorous acid (HOCl), hypobromous acid (HOBr) and hypothiocyanous acid (HOSCN) respectively. These species are antibacterial agents, but misplaced or excessive production is implicated in tissue damage at sites of inflammation. Unlike HOCl and HOBr, which react with multiple targets, HOSCN targets cysteine residues with considerable selectivity. In the light of this reactivity, we hypothesized that Sec (selenocysteine) residues should also be rapidly oxidized by HOSCN, as selenium atoms are better nucleophiles than sulfur. Such oxidation might inactivate critical Sec-containing cellular protective enzymes such as GPx (glutathione peroxidase) and TrxR (thioredoxin reductase). Stopped-flow kinetic studies indicate that seleno-compounds react rapidly with HOSCN with rate constants, k, in the range 2.8×103–5.8×106 M−1·s−1 (for selenomethionine and selenocystamine respectively). These values are ~6000-fold higher than the corresponding values for H2O2, and are also considerably larger than for the reaction of HOSCN with thiols (16-fold for cysteine and 80-fold for selenocystamine). Enzyme studies indicate that GPx and TrxR, but not glutathione reductase, are inactivated by HOSCN in a concentration-dependent manner; k for GPx has been determined as ~5×105 M−1·s−1. Decomposed HOSCN did not induce inactivation. These data indicate that selenocysteine residues are oxidized rapidly by HOSCN, with this resulting in the inhibition of the critical intracellular Sec-dependent protective enzymes GPx and TrxR

Transit of H2O2 across the endoplasmic reticulum membrane is not sluggish

Cellular metabolism provides various sources of hydrogen peroxide (H2O2) in different organelles and compartments. The suitability of H2O2 as an intracellular signaling molecule therefore also depends on its ability to pass cellular membranes. The propensity of the membranous boundary of the endoplasmic reticulum (ER) to let pass H2O2 has been discussed controversially. In this essay, we challenge the recent proposal that the ER membrane constitutes a simple barrier for H2O2 diffusion and support earlier data showing that (i) ample H2O2 permeability of the ER membrane is a prerequisite for signal transduction, (ii) aquaporin channels are crucially involved in the facilitation of H2O2 permeation, and (iii) a proper experimental framework not prone to artifacts is necessary to further unravel the role of H2O2 permeation in signal transduction and organelle biology. © 2016 Elsevier Inc