Anti-GST tag antibody

We studied the absorption, cytotoxicity and oxidative stress markers of Paralytic Shellfish Toxins (PST) from three extracts from Alexandrium catenella and A. ostenfeldii, in middle Oncorhynchus mykiss intestine in vitro and ex vivo preparations. We measured glutathione (GSH) content, glutathione-S transferase (GST), glutathione reductase (GR) and catalase (CAT) enzymatic activity, and lipid peroxidation in isolated epithelium exposed to 0.13 and 1.3 mu M PST. ROS production and lysosomal membrane stability (as neutral red retention time 50%, NRRT50) were analyzed in isolated enterocytes exposed to PST alone or plus 3 mu M of the ABCC transport inhibitor MK571. In addition, the concentration-dependent effects of PST on NRRT50 were assayed in a concentration range from 0 to 1.3 mu M PST. We studied the effects of three different PST extracts on the transport rate of the ABCC substrate DNP-SG by isolated epithelium. The extract with highest inhibition capacity was selected for studying polarized DNP-SG transport in everted and non-everted intestinal segments. We registered lower GSH content and GST activity, and higher GR activity, with no significant changes in CAT activity, lipid peroxidation or ROS level. PST exposure decreased NRRT50 in a concentration-depend manner (IC50 = 0.0045 mu M), but PST effects were not augmented by addition of MK571. All the three PST extracts inhibited ABCC transport activity, but this inhibition was effective only when the toxins were applied to the apical side of the intestine and DNP-SG transport was measured at the basolateral side. Our results indicate that PST are absorbed by the enterocytes from the intestine lumen. Inside the enterocytes, these toxins decrease GSH content and inhibit the basolateral ABCC transporters affecting the normal functions of the cell. Furthermore, PST produce a strong cytotoxic effect to the enterocytes by damaging the lysosomal membrane, even at low, non-neumtoxic concentrations.

Pollution derived from human activities and the arrival of invasive species are common worldwide and affect coastal marine ecosystems negatively, and more especially in a semi-closed sea such as the Mediterranean Sea. The aim of the study was to evaluate oxidative stress biomarkers in the gonadal tissue of the sea urchin Paracentrotus lividus (Lamarck, 1816) sampled in different areas of Sant Antoni de Portmany (Ibiza Island, Spain) with different anthropic activities, and in an area deeply covered by the invasive red algae Lophocladia lallemandii. The densities of P. lividus were higher in the area with the greatest anthropogenic influence, while the area invaded by L. lallemandii showed the lowest density. A significant increase in the activities of the antioxidant enzymes catalase, superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione reductase (GRd) and the phase II detoxifying enzyme glutathione S-transferase (GST) was found in the most impacted area by the human activity. Moreover, malondialdehyde (MDA) and nitrite levels were also increased in the most impacted area. Similarly, the presence of L. lallemandii induced oxidative stress in P. lividus evidenced by a significant increase in all analysed biomarkers. In conclusion, changes in oxidative stress biomarkers are a good proxy to evaluate the impacts induced by anthropogenic activities and by the presence of invasive algae to P. lividus. (C) 2020 Elsevier Ltd. All rights reserved.