Regulation of hepatic glutathione S-transferase expression in flounder

The major glutathione S-transferase isoform of flounder liver, an antigenically related structural homologue of plaice GST-A, also displays mRNA homology. A 901bp cRNA probe for plaice GST-A cross-hybridised to a 1100bp flounder mRNA on northern blot analysis. The plaice antibody and cRNA probes were used to study effects of inducer treatment on GST-A expression in flounder liver. Six days after PAH treatment (3-methylcholanthrene) total hepatic GST activity was halved, levels of GST-A were 80% and GST-A mRNA levels were 25% of controls. A commercial PCB mixture (Aroclor 1254^TM) had little effect on total GST or GST-A levels despite halving GST-A mRNA levels. An epoxide, trans-stilbene oxide induced total GST activity 1·4 fold and GST-A protein levels 1·8-fold and its mRNA levels 3-fold. This reduced expression of the major flounder hepatic GST by agents which induce cytochrome P4501A1 may modulate cytoxicity of environmental pollutants in this species.     

A piscine glutathione S-transferase which efficiently conjugates the end-products of lipid peroxidation

A cDNA clone for glutathione S-transferaseA (GSTA) from plaice (Pleuronectes platessa) was expressed in Eschericia coli (E. coli) and purified to homogeneity by S-hexylglutathione affinity chromatography. When compared to literature values for a variety of purified mammalian GSTs, the heterologously expressed purified plaice enzyme had moderate activity towards the model substrate 1,2-chloro-2,4-dinitrobenzene (CDNB) and exhibited a Km of 2.5 ± 2 mM and Vmax of 30.9 ± 2.3 μmol min⁻¹ mg⁻¹. It had little or no activity towards several other model GST substrates including 1,2-dinitrochloro-4-benzene (DCNB), ethacrynic acid (EA), and p-nitrobenzylchloride (NBC). However plaice GSTA was a relatively efficient catalyst for the conjugation of a series of alk-2-enals and alk-2,4-dienals and also 4-hydroxynonenal. The highest activity observed with this series of substrates was with trans-non-2-enal with a Km of 17.9 ± 2.2 μM and a Vmax of 3.01 ± 0.57 μmol min⁻¹ mg⁻¹. These unsaturated alkenals have been identified in cells and cell extracts as highly toxic products arising from peroxidation of unsaturated fatty acids particularly during periods of oxidative stress. Fish are relatively rich in polyunsaturated fatty acids and thus GSTA mediated conjugation may be an important mechanism for detoxifying peroxidised lipid breakdown products.     

Implications from a field study regarding the relationship between polycyclic aromatic hydrocarbons and glutathione S-transferase activity in mussels

An aluminium smelter on the west coast of Scotland discharges an aqueous effluent containing polycyclic aromatic hydrocarbons (PAHs) at the head of Loch Leven. The loch also supports two mussel (Mytilus edulis) farms. Data are presented on burdens of PAHs in the soft tissues of mussels and the effect of these contaminants on glutathione S-transferase (GST) activity in mussel hepatopancreas. GST activity is shown to be correlated with total PAH burden and also with the concentrations of certain individual PAHs. These field data show that high molecular weight PAHs are closely correlated to GST activity, whereas low molecular weight PAHs are not. This suggests that 5- and 6-ring PAHs have a more pronounced role than 2- to 4-ring compounds in inducing GST activity in mussels from Loch Leven. It is proposed that it may be more appropriate to link GST activity with 5- and 6-ring compounds only, rather than with the total PAH burden.     

GSTA is a major glutathione S-transferase gene responsible for 4-hydroxynonenal conjugation in largemouth bass liver

We have previously shown that largemouth bass (Micropterus salmoides) has a remarkable ability to conjugate 4-hydroxy-2-nonenal (4HNE), a mutagenic and cytotoxic α,β-unsaturated aldehyde produced during the peroxidation of lipids. In addition, we have isolated a glutathione S-transferase cDNA (bass GSTA) that encodes a recombinant protein which is highly active in 4HNE conjugation and structurally similar to plaice (Pleuronectes platessa) GSTA. In the present study, HPLC-GST subunit analysis revealed the presence of at least two major GST isoforms in bass liver, with one peak constituting 80% of the total bass liver GST protein. Liquid chromatography mass spectrometry (LC–MS) and electrospray ionization analysis of the major bass GST subunit yielded a molecular weight of 26,396 kDa. Endo-proteinase Lys-C digestion and Edman degradation protein sequencing of this GST peak demonstrated that this protein was encoded by bass GSTA. Analysis of genomic DNA fragments isolated by nested PCR indicated the presence of a GST gene cluster in bass liver that contained GSTA, and was similar to a GST gene cluster characterized by Leaver et al., in plaice. Collectively, our data indicates the presence of a major GST in bass liver involved in the protection against oxidative stress. This GST is part of a gene cluster that may be conserved in certain freshwater and marine fish.     

Effects of three fungicides alone and in combination on glutathione S-transferase activity (GST) and cytochrome P-450 (CYP 1A1) in the liver and gill of brown trout (Salmo trutta)

In order to evaluate the gill glutathione S-transferase (GST) activity as a biomarker of effect of fungicide exposure in juvenile brown trout (Salmo trutta), the fungicides propiconazole {(R,S)-1-[2-(2,4-diclophenyl)-4-propyl-1,3-dioolan-2-ylmetyl]-1H-1,2,4-triazole} and fenpropimorph {(±)-cis-4-[3-(4-tert-butylphenyl)-2-metyl propyl]-2,6 dimetylmorfolinc} were administrated in the water separately and together in a static system (80 μg/l for each pesticide) for 5 days. The combined fungicides gave a significant decrease in gill GST activity towards 1-chloro-2,4-dinitrobenzene (CDNB), whilst hepatic GST-activity was not significantly changed. Furthermore, continuous exposure to 540 ug/l thiabendazole{ 2-(thiazol-4′-yl)benzimidazole} in a flow-through system for 4 days significantly increased the gill glutathione S-transferase (GST) activity towards CDNB, whilst hepatic GST and cytochrome P450 (CYP 1A) activities were not increased by the treatment.     

The distribution and population dynamics of O-group plaice (Pleuronectes platessa L.) on nursery grounds in the Firth of Forth

Midsummer (1 August) population estimates of about 2 million O-group plaice (Pleuronectes platessa L.) were derived for sandy bays around the Firth of Forth in 1979–1980. This is an order of magnitude less than similar estimates made for the Clyde Sea Area in 1973–1974. Autumn population estimates of 0·4–1·0 million fish were comparable to estimates by the Ministry of Agriculture, Fisheries and Food for the area between the Scottish border and Flamborough Head (2·3 million for 1970 and 1973) which represented 4·8% (1973) to 5·3% (1970) of the total number of O-group fish on the English east coast.Largo Bay was the most important nursery area holding 25% of the total population. It is particularly well situated to receive newly metamorphosed plaice carried in water currents along the north side of the Forth from the spawning ground off Fife Ness. Plaice in the Forth are mainly distributed on fine to medium sandy beaches (186–480 μm), the mean number per haul in midsummer (D) being correlated with the median diameter (m.d. in μm) of the low water sediments by the equation: D=−45·7666+0·2327 m.d. (n=11,r=0·68,P<0·02 but>0·01).The shallow inshore water in sandy bays in the outer Firth was well mixed and more marine than estuarine (27·7–35·0‰). The correlation coefficient between fish density and water temperature was low, while that with salinity (S‰) was: D=6·1618+0·2238S (n=23,r=0·62,P<0·005).Regression analysis demonstrated that the relationship between the instantaneous mortality rate (Z) and the initial population density (D_p) was: Z×100=0·7480+0·0546d_p (n=12,r=0·87,P<0·001).The mean mortality rate for the O-group plaice in the Forth nursery areas was 53% month⁻¹.     

The effect of salinity on growth and weight loss of juvenile plaice (Pleuronectes platessa, L): An experimental test

Previous population estimates of the 0+ plaice (Pleuronectes platessa L.) in the Firth of Forth, east central Scotland, did not take account of the Forth estuary west of the Forth bridges. Previous work found plaice in the estuary grew as fast as, or faster than, the outer firth plaice. It was hypothesised that salinity may affect growth rates of early 0+ plaice. This hypothesis was tested in a laboratory experiment, by exposing juvenile plaice to three different, but naturally — experienced by the juveniles, salinities; 25, 30 and 35. Plaice fed a minimum ration did not grow in length. Mean weight decreased at all three salinities, however, the lowest weight loss was found at the lowest salinity (25) and the highest weight loss was found at the highest salinity (35). The minimum feeding ration was halted and plaice were then fed ad libitum. Consumption rates were not significantly different during the ad libitum feeding, while significant differences in mean weight change were found between the highest and lowest salinities.     

凡纳滨对虾造血激素的组织分布及细胞定位分析

造血激素（astakine）是一种具有促进造血组织细胞分化和增殖作用的甲壳动物细胞因子,参与甲壳动物的造血活动,对于只依赖于非特异性免疫的甲壳动物抗感染能力有重要的意义。本实验室前期成功克隆凡纳滨对虾造血激素（LvAST）的基因Lvast。为了进一步了解其功能,本文通过实时荧光定量PCR技术检测了Lvast表达的组织分布,同时还用Dlight标记抗LvAST抗体进行了LvAST在对虾细胞和组织中的免疫荧光定位分析。荧光定量PCR测定结果说明,Lvast主要在肝胰腺、鳃和肌肉中大量表达,而淋巴器官和血淋巴细胞中表达量较低;免疫荧光结果显示,LvAST可同时存在于血淋巴细胞内和血淋巴细胞膜表面,不同的血淋巴细胞表面的LvAST分布呈现出明显差异,可能具有重要的血细胞分类意义。LvAST的蛋白在肝胰腺、鳃、肌肉、淋巴器官等组织中均有分布,并且肝胰腺组织中分布较多,淋巴器官组织中分布较少。     

The effect of salinity on growth and weight loss of juvenile plaice (Pleuronectes platessa,L): An experimental test

Previous population estimates of the 0+ plaice (Pleuronectes platessa L.) in the Firth of Forth, east central Scotland, did not take account of the Forth estuary west of the Forth bridges. Previous work found plaice in the estuary grew as fast as, or faster than, the outer firth plaice. It was hypothesised that salinity may affect growth rates of early 0+ plaice. This hypothesis was tested in a laboratory experiment, by exposing juvenile plaice to three different, but naturally — experienced by the juveniles, salinities; 25, 30 and 35. Plaice fed a minimum ration did not grow in length. Mean weight decreased at all three salinities, however, the lowest weight loss was found at the lowest salinity (25) and the highest weight loss was found at the highest salinity (35). The minimum feeding ration was halted and plaice were then fed ad libitum. Consumption rates were not significantly different during the ad libitum feeding, while significant differences in mean weight change were found between the highest and lowest salinities.     

Changes in the spatial distribution of North Sea plaice (Pleuronectes platessa) and implications for fisheries management

To protect the main nursery area of plaice, an area called the ‘Plaice Box’ was closed to trawl fisheries with large vessels in 1989, with the expectation that recruitment, yield and spawning stock biomass would increase. However, since then the plaice population has declined and the rate of discarding outside the Plaice Box has increased, suggesting an offshore shift in spatial distribution of juvenile plaice. Using research vessel survey data collected since 1970, the change in distribution of juvenile age groups was analysed in relation to the distance to the coast. Further, a comparison of the distribution of different length classes of plaice between three historic periods was made (1902–1909; 1983–1987; 1999–2003). A shift towards deeper water of larger-sized plaice (20–39 cm) is apparent already before the 1980s and may be related to the decrease in the number of competitors or predators. An offshore shift in the distribution of young plaice occurred in the 1990s most likely in response to higher water temperatures that may have exceeded the maximum tolerance range or increased the food requirements above the available food resources. A decrease in competition with larger plaice offshore, possibly in combination with increased inshore predation by cormorants and seals, may also have played a role. The offshore shift in distribution has reduced the effectiveness of the Plaice Box as a technical measure to protect the under-sized plaice from discarding, since an increased proportion of the population of undersized plaice is moving to the more heavily exploited offshore areas.     

Biomarkers of biochemical and cellular stress in Carcinus maenas: an in situ field study

Biomarkers are useful diagnostic tools for identifying exposure to physical, chemical, and environmental stresses. In this study a suite of techniques—measurements of metallothionein, lysosomal integrity, and osmoregulatory ability—have been used in an attempt to provide an in situ assessment of environmental quality. Five estuarine sites were studied, two clean sites, two urban sites, and one intermediate. Shore crabs, Carcinus maenas, were collected and haemolymph taken for the lysosomal and osmoregulatory assays. Animals were killed and tissue preserved for metallothionein analysis. Lysosomal integrity was significantly different in crabs from clean sites and the urban sites (n = 40, p < 0.05). The potential value of the biomarker approach is that it could provide data for impact assessment and is applicable in the field.     

An inverse modeling study in Fram Strait. Part II: Water mass distribution and transports

A water-mass analysis is carried out in Fram Strait, between 77.15 and 81.15°N, based on three-dimensional large-scale potential temperature and salinity distributions reconstructed from the MIZEX 84 hydrographic data collected in summer 1984. Combining these distributions with the geostrophic flow field derived from the same data in a companion paper (Schlichtholz and Houssais, 1999), the heat, fresh water and volume transports are estimated for each of the water masses identified in the strait. Twelve water masses are selected based on their different origins. Among them, the Polar Water (PW) enters Fram Strait from the Arctic Ocean both over the Greenland Slope and over the western slope of the Yermak Plateau. In the Atlantic Water (AW) range, four modes with distinct geographical distributions are indentified. In the Deep Water range, the Eurasian Basin Deep Water (EBDW) is confined to the Lena Trough and to the Molloy Deep area where it is involved in a cyclonic circulation. The warm and shallower mode of the Norwegian Sea Deep Water (NSDW), concentrated to the west, is mainly seen as an outflow from the Arctic Ocean while the cold and deeper mode, essentially observed to the east, enters the strait from the Greenland Sea. Apart from the EBDW, there is a tendency for all water masses of polar origin to flow along the Greenland Slope. The two most abundant water masses, the AW and the NSDW, occupy as much as 67% of the total water volume. The southward net transport of PW through Fram Strait is about 1 Sv at 78.9°N. At the same latitude, the net transport of AW is southward and equal to about 1.7 Sv. Only the transport of the warm mode (AWw) is northward, amounting to 0.2 Sv. The overall net outflow of the Deep Waters to the Greenland Sea is about 2.6 Sv. Two upper water masses, the fresh (AWf) and the cold (AWc) mode of the AW, and one deep-water mass, the NSDW, appear to be produced in the strait, with production rates, between 77.6 and 79.9°N, of about 0.2, 1.0 and 1.7 Sv, respectively. A southward net fresh-water transport through the strait of about 2000 km³ yr⁻¹ (relative to a salinity of 34.93) is mainly due to the PW. The net heat transport relative to −0.1°C is northward, but undergoes a rapid northward decrease, suggesting an area-averaged surface heat loss of 50–100 W m⁻² in the strait.     

水下溢油油滴粒径分布实验研究

Oil droplet size distribution(ODSD) plays a critical role in the rising velocity and transport of oil droplets in subsurface oil releases. In this paper, subsurface oil release experiments were conducted to study ODSD under different experimental conditions in a laboratory water tank observed by two high-speed cameras in March and April 2017. The correlation formulas Oh=10.2 Re~(–1) and Oh=39.2 Re~(–1)(Re represents Reynolds number and Oh represents Ohnesorge number) were established to distinguish the boundaries of the three instability regimes in dimensionless space based on the experimental results. The oil droplet sizes from the experimental data showed an excellent match to the Rosin–Rammler distribution function with determination coefficients ranging from 0.86 to 1.00 for Lvda 10-1 oil. This paper also explored the influence factors on and change rules of oil droplet size. The volume median diameter d50 decreased steadily with increasing jet velocity, and a sharp decrease occurred in the laminar-breakup regime. At Weber numbers(We) 100, the orifice diameter and oil viscosity appeared to have a large influence on the mean droplet diameter. At 100We1 000, the oil viscosity appeared to have a larger influence on the relative mean droplet diameter.     

Modelling the spatial distribution of plaice (Pleuronectes platessa), sole (Solea solea) and thornback ray (Raja clavata) in UK waters for marine management and planning

Species distribution maps are needed for ecosystem-based marine management including the development of marine spatial plans. If such maps are based on predictive models then modelling procedures should aim to maximise validation success, and any uncertainty in the predictions needs to be made explicit. We developed a predictive modelling approach to produce robust maps of the distributions of selected marine species at a regional scale. We used 14 years of survey data to map the distributions of plaice, sole and thornback ray in three hydrographic regions comprising parts of the Irish Sea, Celtic Sea and the English Channel with the help of the hybrid technique regression kriging, which combines regression models with geostatistical tools. For each species–region combination we constructed logistic Generalized Linear Models (GLMs) based on presence–absence data using the environmental variables: depth, bottom temperature, bed shear stress and sediment type, as predictors. We selected GLMs using the mean squared error of prediction (MSEP) estimated by cross-validation then conducted a geostatistical analysis of the residuals to incorporate spatial structure in the predictions. In general, we found that species occurrence was positively related to shallow areas, a bed shear stress of between 0 and 1.5 N/m², and the presence of sandy sediment. Predicted species occurrence probabilities were in good agreement with survey observations. This modelling framework selects environmental models based on predictive ability and considers the effect of spatial autocorrelation on predictions, together with the simultaneous presentation of observations, associated uncertainties, and predictions. The potential benefit of these distribution maps to marine management and planning is discussed.     

Evidence from heterologous expression of glutathione S-transferases A and A1 of the plaice (Pleuronectes platessa) that their endogenous role is in detoxification of lipid peroxidation products

cDNA clones for glutathione S-transferases A (GST-A) and A1 (GST-A1) from plaice (Pleuronectes platessa) were expressed as N-terminally 6XHis tagged proteins in Escherichia coli and purified to homogeneity from Ni-NTA silica. GST-A was an efficient catalyst for conjugation of unsaturated alkenals derived from peroxidation of polyunsaturated fatty acids with the highest activity observed with trans-non-2-enal (8 micromol min(-1) mg(-1)). GST-A1 was a very efficient Se-independent glutathione peroxidase with an activity towards cumene hydroperoxide of 25 micromol min(-1) mg(-1). Although the enzymes exhibited moderately high activities towards the model substrate 1-chloro-2,4-dinitrobenzene (CDNB) they exhibited little or no activity towards other common prototypical xenobiotic substrates. Together with data for ontogeny, tissue distribution and inducibility of these enzymes, we contend that a primary function of these enzymes is protection from the harmful effects of lipid peroxidation products generated naturally or exacerbated by xenobiotic exposure.