Comparison of cytochrome P450 in three butterflyfish species: Ecological implications of elevated CYP2B and CYP3A in Chaetodon capistratus

The relationship between cytochrome P450 and feeding on terpenoid-rich gorgonian corals was investigated in a species of tropical butterflyfish and compared with two other sympatric congeners that do not feed on gorgonians. Fish were collected from non-polluted waters in Belize and the levels of two cytochrome P450 isozymes (CYP2B and CYP3A) were immunoquantitated in addition to quantification of total P450. Chaetodon capistratus regularly feeds on gorgonian corals and has higher levels of total hepatic microsomal cytochrome P450 than C. ocellatus or C. striatus. The content of hepatic P450 (0.588–0.794 nmol mg⁻¹) in C. capistratus is among the highest ever reported in teleosts from non-polluted waters and is significantly greater than detected in C. ocellatus or C. striatus. Chaetodon capistratus also had a larger hepatic index (g liver per g fish) and more microsomal protein (mg protein per g liver), factors that translate into 3.3- to 8-fold more total P450 per g fish. Sexual differences in total P450 were observed between male and female C. capistratus, but not among the other species. The contents of proteins detected by immunoassay with polyclonal anti-scup P450B (CYP2B) and anti-human P4503A (CYP3A) were 2- to 10-fold and 2- to 20-fold greater, respectively, in C. capistratus than in the congeneric species. CYP2 and CYP3 gene families in mammals are thought to have evolved partially in response to dietary allelochemicals. These results suggest that these P450 isozymes may also be important in marine teleosts that feed on terpenoid-rich prey.     

Response of the blue mussel Mytilus edulis L. following exposure to PAHs or contaminated sediment

The polycyclic aromatic hydrocarbons fluoranthene and benzo(a)pyrene significantly reduced the feeding rate of mussels. For both compounds the tissue concentration resulting in a 50% reduction of the clearance rate (TEC₅₀) was calculated. At high tissue concentrations both aromatic compounds reduced the tolerance of mussels to aerial exposure, whereas at low tissue concentrations an improved response was noticed. The activities of the antioxidant enzymes Superoxide dismutase (SOD) and catalase were elevated only at low tissue concentrations of fluoranthene and benzo(a)pyrene. At the highest measured tissue concentrations the activity of both enzymes was reduced, possibly due to a narcotic effect. The reproductive success rate of mussels appeared to be affected negatively by the investigated hydrocarbons. The results of a pilot experiment indicate that mussels can be used also for the testing of contaminated sediments.     

Tectonostratigraphy and sedimentary architecture of rift basins, with reference to the northern North Sea

A tectonostratigraphic model for the evolution of rift basins has been built, involving three distinct stages of basin development separated by key unconformities or unconformity complexes. The architecture and signature of the sediment infill for each stage are discussed, with reference to the northern North Sea palaeorift system. The proto-rift stage describes the rift onset with either doming or flexural subsidence. In the case of early doming, a proto-rift unconformity separates this stage from the subsequent main rift stage. Active stretching and rotation of fault blocks during the rift stage is terminated by the development of the syn-rift unconformity. Where crustal separation is accomplished, a break-up unconformity commonly marks the boundary to the overlying thermal relaxation or post-rift stage. Tabular architectures, thickening across relatively steep faults, characterize the proto-rift stage. Syn-rift architectures are much more variable. Depending on the ability of the sediment supply to fill the waxing and waning accommodation created during rotation and subsidence, one-, two- or three-fold lithosome architectures are likely to develop. During the post-rift stage, an early phase with coarse clastic infilling of remnant rift topography often precedes late stage widening of the basin and filling with fine-grained sediments.     

Buckling of circular, solid and annular plates with an intermediate circular support

The title problem is solved in the case where uniform applied loading is present at the plate outer boundary. Two independent solutions are obtained: the optimized Rayleigh–Ritz method and the finite element algorithmic procedure. Good engineering agreement is shown to exist. The proposed analytical procedure is convenient from a designer's viewpoint. Admittedly exact analytical solutions can be found in terms of Bessel functions, the procedure being rather complex from an engineering viewpoint.     

Tectonics of an extinct ridge-transform intersection,Drake Passage (Antarctica)

New swath bathymetric, multichannel seismic and magnetic data reveal the complexity of the intersection between the extinct West Scotia Ridge (WSR) and the Shackleton Fracture Zone (SFZ), a first-order NW-SE trending high-relief ridge cutting across the Drake Passage. The SFZ is composed of shallow, ridge segments and depressions, largely parallel to the fracture zone with an `en echelon' pattern in plan view. These features are bounded by tectonic lineaments, interpreted as faults. The axial valley of the spreading center intersects the fracture zone in a complex area of deformation, where N120° E lineaments and E–W faults anastomose on both sides of the intersection. The fracture zone developed within an extensional regime, which facilitated the formation of oceanic transverse ridges parallel to the fracture zone and depressions attributed to pull-apart basins, bounded by normal and strike-slip faults.On the multichannel seismic (MCS) profiles, the igneous crust is well stratified, with numerous discontinuous high-amplitude reflectors and many irregular diffractions at the top, and a thicker layer below. The latter has sparse and weak reflectors, although it locally contains strong, dipping reflections. A bright, slightly undulating reflector observed below the spreading center axial valley at about 0.75 s (twt) depth in the igneous crust is interpreted as an indication of the relict axial magma chamber. Deep, high-amplitude subhorizontal and slightly dipping reflections are observed between 1.8 and 3.2 s (twt) below sea floor, but are preferentially located at about 2.8–3.0 s (twt) depth. Where these reflections are more continuous they may represent the Mohorovicic seismic discontinuity. More locally, short (2–3 km long), very high-amplitude reflections observed at 3.6 and 4.3 s (twt) depth below sea floor are attributed to an interlayered upper mantle transition zone. The MCS profiles also show a pattern of regularly spaced, steep-inclined reflectors, which cut across layers 2 and 3 of the oceanic crust. These reflectors are attributed to deformation under a transpressional regime that developed along the SFZ, shortly after spreading ceased at the WSR. Magnetic anomalies 5 to 5 E may be confidently identified on the flanks of the WSR. Our spreading model assumes slow rates (ca. 10–20 mm/yr), with slight asymmetries favoring the southeastern flank between 5C and 5, and the northwestern flank between 5 and extinction. The spreading rate asymmetry means that accretion was slower during formation of the steeper, shallower, southeastern flank than of the northwestern flank.     

Large aperture seismic imaging at a convergent margin: Techniques and results from the Costa Rica seismogenic zone

In March and April 1995 a cooperative German, Costa Rican, and United States research team recorded onshore-offshore seismic data sets along the Pacific margin of Costa Rica using the R/V Ewing. Off the Nicoya Peninsula we used a linear array of ocean bottom seismometers and hydrophones (OBS/H) with onshore seismometers extending across much of the isthmus. In the central area we deployed an OBS/H areal array consisting of 30 instruments over a 9 km by 35-km area and had land stations on the Nicoya Peninsula adjacent to this marine array and also extending northeast on the main Costa Rican landmass. Our goal in these experiments was to determine the crustal velocity structure along different portions of this convergent margin and to use the dense instrument deployments to create migrated reflection images of the plate boundary zone and the subducting Cocos Plate. Our specific goal in the central area was to determine whether a subducted seamount is present at the location of the 1990, M 7 earthquake off the Nicoya Peninsula and can thus be linked to its nucleation. Subsequently we have processed the data to improve reflection signals, used the data to calculate crustal velocity models, and developed several wide-aperture migration techniques, based on a Kirchhoff algorithm, to produce reflection images. Along the northern transect we used the ocean bottom data to construct a detailed crustal velocity model, but reflections from the plate boundary and top and bottom of the subducting Cocos plate are difficult to identify and have so far produced poor images. In contrast, the land stations along this same transect recorded clear reflections from the top of the subducting plate or plate boundary, within the seismogenic zone, and we have constructed a clear image from this reflector beneath the Nicoya shelf. Data from the 3-D seismic experiment suffer from high-amplitude, coherent noise (arrivals other than reflections), and we have tried many techniques to enhance the signal to noise ratio of reflected arrivals. Due to the noise, an apparent lack of strong reflections from the plate boundary zone, and probable structural complexity, the resulting 3-D images only poorly resolve the top of the subducting Cocos Plate. The images are not able to provide compelling evidence of whether there is a subducting seamount at the 1990 earthquake hypocenter. Our results do show that OBS surveys are capable of creating images of the plate boundary zone and the subducting plate well into the seismogenic zone if coherent reflections are recorded at 1.8 km instrument spacing (2-D) and 5 km inline by 1 km crossline spacing for 3-D acquisition. However, due to typical high amplitude coherent noise, imaging results may be poorer than expected, especially in unfavorable geologic settings such as our 3-D survey area. More effective noise reduction in acquisition, possibly with the use of vertical hydrophone arrays, and in processing, with advanced multiple removal and possibly depth filtering, is required to achieve the desired detailed images of the seismogenic plate boundary zone.     

Biochemical and histochemical responses to environmental contaminants in clam, Tapes philippinarum, transplanted to different polluted areas of Venice Lagoon, Italy.

In this study the clam, Tapes philippinarum, a filter-feeding bivalve living in soft bottoms, was used to investigate the water/sediment pollution in Venice Lagoon (Venice, Italy), a heavily urbanised and industrialised area. To this end, clams collected from indigenous populations in a clean farming area (Val Dogà) were transplanted into a relatively low contaminated site (Palude della Rosa) and in a heavily polluted area (Porto Marghera), for 5 weeks. A range of possible induced stress indices were measured, on the basis of potential toxicity mechanisms, including specific enzymatic activities [Bap hydroxylase and NADPH cytochrome c reductase, superoxide dismutase, catalase, glutathione peroxidase and propionaldehyde dehydrogenase (PDH)] and general cellular responses (lysosomal latency). Moreover, chemical analyses (polychlorinated biphenyls, polycyclic aromatic hydrocarbon, DDTs) were carried out on sediments and animal tissues at the beginning and end of the transplantation experiments. The chemical results show an active bio-accumulation activity from sediment in clams translocated to the most polluted area. Correspondingly, biological data indicate marked effects on PDH activity and latency in T. philippinarum from the same site. In contrast, no similar pattern has been observed in the other considered indices, possibly due to antagonistic effects of the complex contaminant mixture present in the environment.     

Relation between responses in the neutral red retention test and the comet assay and life history parameters of Daphnia magna.

Responses of the neutral red retention (NRR) assay as test for lysosomal stability and the comet assay as test for DNA integrity were measured in the water flea, Daphnia magna, and compared with mortality and effects on population growth rate during short- or long-term exposure to seven different toxicants. The NRR test and the comet assay were performed with fresh preparations of pieces of tissue from the digestive tract or with cell preparations from whole daphnias. Five toxicants caused responses of the NRR test or the comet assay after short-term exposure at concentrations below the acute toxicity level. Preliminary results of long-term exposure experiments suggest that these biomarker responses can be related to chronic effects on survival and/or reproduction of D. magna. This type of research should provide the basis for future use of the NRR test and the comet assay as early warning biomarkers for effects of toxicants on Daphnia populations.     

The requirements for stable metallothionein clusters examined using synthetic lobster domains.

Metallothioneins (MTs) are small, cysteine-rich proteins which detoxify xenobiotic metals such as cadmium (Cd) and mercury (Hg). In crustaceans and mammals they consist of two independent domains which are folded around metal-thiolate clusters. MT clusters of different origins, exhibiting distinct, highly conserved cysteine positions on their sequences, show differences in metal-cysteine coordination and reactivity. Lobster-MT, containing two Cd3 beta domains, is an important model for structure-function relationships among the clusters. The influence of (1) the position of the cysteine residues and (2) steric and electrostatic effects of neighboring amino acids on the folding and stability of MT cluster were investigated. Thus, the native lobster beta C and beta N domains (each having nine cysteines and binding three M2+ ions) and a modified domain Cd3 beta C-->N, in which the cysteines of the C-terminal domain were relocated to match the positions of those in the N-terminal domain, were chemically prepared and characterized. The synthetic native domains (Cd3 beta C and Cd3 beta N) were found to exhibit spectroscopic properties, metal-binding affinities and kinetic reactivity similar to the holo-protein. However, the modified Cd3 beta C-->N domain was unusually reactive and in the presence of Chelex, metal chelation resin, aggregated to a Cd5(beta C-->N)2 dimer, which exhibited unusual structure as observed by its 113Cd-nuclear magnetic resonance. These differences in structure and reactivity demonstrated that the requirements for formation of a stable Cd3S9 beta-cluster are more stringent than simply the sequential positions of the cysteines along the peptide chain and must include interactions involving neighboring, noncysteine amino acids.