Analysis of medaka cytochrome P450 3A homotropic and heterotropic cooperativity

We have previously demonstrated that medaka CYP3A is associated with metabolism of several endobiotics including steroids and bile acids. In this study, we demonstrate that medaka CYP3A catalysis exhibits unusual kinetic behaviors consistent with allosteric interaction which cannot be described by hyperbolic kinetic models. Using 7-benzyloxy-4-(trifluoromethyl)-coumarin (BFC) and nonylphenol as CYP3A substrates, we describe both homotropic and heterotropic cooperative interactions. Given the role of CYP3A in maintaining the homeostatic balance for numerous endobiotics, enzymatic activation/inhibition by endocrine disruptors (EDCs) represents a putative (non-genomic) mechanism for endocrine disruption.     

Imaging of Buried Archaeological Materials: The Reflection Properties of Archaeological Wood

Effective marine archaeological site management demands detailed information on not only the spatial distribution of artefacts but also the degradation state of the materials present. Although sonar methods have frequently been used in an attempt to detect buried wooden shipwrecks they are currently unable to indicate their degradation state. To assess the sensitivity of acoustic measurements to changes in the degradation state of such material, and hence the potential for sonars to quantify degradation, laboratory measurements of compressional wave velocity, as well as bulk density for oak and pine samples, in varying states of decay, were undertaken. These data enabled the calculation of theoretical reflection coefficients for such materials buried in various marine sediments. As wood degrades, the reflection coefficients become more negative, resulting in the hypothesis that the more degraded wood becomes, the easier it should be to detect. Typical reflection coefficients of the order of −0.43 and −0.52 for the most degraded oak and pine samples in sand are predicted. Conversely, for wood exposed to seawater the predicted reflection coefficients are large and positive for undegraded material (0.35 for oak, 0.18 for pine) and decrease to zero or slightly below for the most degraded samples. This indicates that exposed timbers, when heavily degraded, can be acoustically transparent and so undetectable by acoustic methods. Corroboration of these experimental results was provided through comparison with high resolution seismic reflection data that has been acquired over two shipwrecks.     

Management of a shallow temperate estuary to control eutrophication: The effect of hydrodynamics on the system’s nutrient loading

The Mondego estuary, a shallow warm-temperate intertidal system located on the west coast of Portugal, has for some decades been under severe ecological stress, mainly caused by eutrophication. Water circulation in this system was, until 1998, mainly dependent on tides and on the freshwater input of a small tributary artificially controlled by a sluice. After 1998, the sluice opening was effectively minimised to reduce the nutrient loading, and the system hydrodynamics improved due to engineering work in the upstream areas. The objective of the present study was to evaluate the effect of the mitigation measures implemented in 1998. Changes to the hydrodynamics of the system were assessed using precipitation and salinity data in relation to the concentrations of dissolved inorganic nutrients, as well as the linkage between dissolved N:P ratios and the biological parameters (phytoplankton chlorophyll a concentrations, green macroalgal biomass and seagrass biomass). Two distinctive periods were compared, over a ten year period: from January 1993 to January 1997 and from January 1999 until January 2003. The effective reduction in the dissolved N:P atomic ratio from 37.7 to 13.2 after 1998 is a result of lowered ammonia, but not the oxidised forms of nitrogen (nitrate plus nitrite), or increased concentrations of dissolved inorganic phosphorus. Results suggest that the phytoplankton is not nutrient limited, yet maximum and mean biomass of green macroalgae was reduced by one order of magnitude after the mitigation measures. This suggests that besides lowering the water residence time of the system, macroalgal growth became nitrogen limited. In parallel to these changes the seagrass-covered area and biomass of Zostera noltii showed signs of recovery.     

Longitudinal strength of a high-speed ferry

The longitudinal strength of the high-speed ferry was investigated by subjecting the ship's hull girder to long-term loads obtained from a frequency-domain panel code. Prior to the statistical analysis, linearly computed transfer functions were corrected for nonlinear effects, yielding two sets of transfer functions valid for different wave amplitudes. One set corresponded to the hogging condition; the other set, to the sagging condition. Two regular equivalent design waves were specified that resulted in loads representing the most severe global design load conditions. The still-water loading condition, yielding a still-water vertical bending moment in hogging, was superimposed on the wave-induced loads to obtain the total (design) loads in hogging. For the sagging condition only, additional impact-related loads were superimposed to obtain the total (design) loads in sagging. A finite element model of the ship's structure was subjected to pressure distributions according to the two regular design waves. For comparison with classification society rule values, a simple beam theory strength analysis of the ship's midship section was performed first, and then another finite element analysis was carried out, whereby the imposed loads were tuned to the rule values of vertical bending moments. Rule-based magnitudes of nominal maximum longitudinal stress deviated significantly (25–39%) from comparable stresses obtained by the panel code based finite element analysis. However, stresses obtained from the rule-based finite element analysis agreed more favorably, especially in hogging. In the uppermost deck, for example, the panel code based compressive stress was only 9% larger than the comparable stress from the rule-based finite element analysis.     

Development of two novel CYP-antibodies and their use in a PCB exposure experiment with Mytilus edulis

In an attempt to learn more about the cytochrome P450 (CYP) system of mussels, we used protein databases and alignment software to extract highly conserved CYP sequences. From these alignments synthetic peptides were produced and used for rabbit immunisation, which yielded polyclonal antibodies against the CYP families 2 and 4. The antibodies were evaluated with Western Blot and ELISA assays, using digestive gland microsomal samples from the mussel Mytilus edulis. Western Blots revealed immunoreactions for both antibodies. The anti-CYP2 sequence rendered one major immunopositive protein of ≈49 kDa size, and weak signals for proteins of ≈41 and 56 kDa size. The anti-CYP4 sequence rendered two major bands of ≈56 and 59 kDa size, and also a weak immunoreaction with a protein of ≈43 kDa size. ELISA rendered only weak signals even with a 1:50 dilution of IgG-purified serum. A 10-day exposure to Aroclor 1254 did not appear to affect any of the immunopositive proteins, while total PCBs in soft bodies increased from 14–40 ng/g DW in controls to 373–638 ng/g DW in exposed mussels.     

Simulating a heavy metal spill under estuarine conditions: Effects on the clam Scrobicularia plana

We describe the effect of heavy metals Zn, Cd, Pb and Cu on the induction of methallothioneins on the clam Scrobicularia plana along a salinity gradient simulated under laboratory conditions. The clams were exposed to constant heavy metal concentrations in a dynamic estuary simulator during a 15-day assay to investigate possible induction of metal-binding proteins in them. The concentration of heavy metals in water was analysed. Clams were analysed for methallothionein concentrations. The speciation of Zn, Cd, Pb and Cu along the salinity gradient was modelled. Zn showed the highest concentrations and its prevalent species was the free ion. Intersite differences have been observed in methallothionein concentration and related to the salinity gradient. It seems that synthesis of methallothioneins is the result of physiological forces acting in concert with the changes in the chemical speciation of metals, owing to the trace metals uptake is controlled by means of an interaction of physiology and physicochemistry.     

Response of the barrier island coastal region of southwestern Nigeria to climate and non-climate forcing

Despite threats emanating from the influence of climate and non-climate forcing on the barrier island coastal region of southwestern Nigeria, the extent of the coastal erosion is poorly understood. We report evidence of coastal erosion and sediment accumulation in the region over a 34-year period (1973–2017), using Landsat imagery at intervals of approximately six years. Landsat image corrections and various water-extraction algorithms were used to systematically delineate coastal erosion and accumulation in the area. The region was subdivided into western and eastern subregions separated by Lagos Harbour. In the west, erosion took place during the periods 1973–1979, 1979–1984, 1990–1999 and 2005–2011, whereas in the east, erosion occurred during 1973–1979, 1990–1999 and 1999–2005. Coastal sediment accumulation occurred in the east during 1979–1984, 1984–1990, 2005–2011 and 2011–2017, whereas gains in the west occurred during 1984–1990, 1999–2005 and 2011–2017. The study revealed substantial net erosion of 1 228.1 ha in the region as a whole, over the full period. Sediment accumulation accompanying the coastal erosion appears to be linked to longshore drift. Erosion between 1973 and 2011 was probably attributable to climate change (storms and tidal conditions), longshore drift, the inflow and outflow of water at Lagos Harbour, coastal morphology and, possibly, human impacts. However, the coastal changes between 2011 and 2017 were more obviously associated with human activities, such as development of the Eko Atlantic construction project. Coastal surveillance, together with the use of environmentally sensitive protective measures, could possibly help to reduce coastal erosion in the region. Careful coastal management practices, including artificial nourishing and the installation of resilient structures (e.g. seawalls), should be undertaken to protect human settlements that are already at risk from sea-level rise.     

Retention and fate of experimentally added mercury in a Massachusetts salt marsh treated with sewage sludge

A simple, rapid, precise and reproducible procedure is described for sample preparation, extraction and measurement of total mercury in peaty sediments, plants and animal tissues. Various amounts of a heavy metal containing dried sewage sludge were applied to experimental plots in a New England salt marsh. This treatment resulted in elevation of mercury levels in the surface 5 cm of the soil. Discrepancies in the bulk density of marsh sediments corresponded with interparticle spaces and detritus/mineral ratios in the peat matrix. Mass balance calculations showed that wetlands covered with tall Spartina alterniflora lost mercury considerable faster than higher marsh locations which retained virtually all mercury added. The shortest halflife of mercury calculated was four years. Grain size analyses of peat and sewage sludge suggested that mercury in the higher intertidal range remained associated with sludge components. Biochemical and physical processes affecting the dissipation of mercury from creek side sediments are discussed.