Changes in the antioxidant metabolism in the embryonic development of the common South American toad Bufo arenarum: differential responses to pesticide in early embryos and autonomous-feeding larvae.

Amphibians may be critically challenged by aquatic contaminants during their embryonic development. Many classes of compounds, including organophosphorus pesticides, are able to cause oxidative stress that affects the delicate cellular redox balance regulating tissue modeling. We determined the progression of antioxidant defenses during the embryonic development of the South American common toad, Bufo arenarum. Superoxide dismutase (SOD) and catalase (CAT) activities were high in the unfertilized eggs, and remained constant during the first stages of development. SOD showed a significant increase when the gills were completely active and opercular folds began to form. Reductase (GR) activity was low in the oocytes and increased significantly when gills and mouth were entirely developed and the embryos presented a higher exposure to pro-oxidant conditions suggesting an environmental control. Reduced glutathione (GSH) content was also initially low, and rose continuously pointing out an increasing participation of GSH-related enzymes in the control of oxidative stress. GSH peroxidases and GSH-S-transferases showed relatively high and constant activities, probably related to lipid peroxide control. B. arenarum embryos have plenty of yolk platelets containing lipids, which provide the energy and are actively transferred to the newly synthesized membranes during the early embryonic development. Exposure to the pro-oxidant pesticide malathion during 48 h did not significantly affect the activity of antioxidant enzymes in early embryos, but decreased the activities of CAT, GR, and the pool of GSH in larvae. Previous work indicated that lipid peroxide levels were kept low in malathion-exposed larvae, thus we conclude that oxidative stress is overcome by the antioxidant defenses. The increase in the antioxidant metabolism observed in the posthatching phase of development of B. arenarum embryo, thus constitutes a defense against natural and human-generated pro-oxidants present in the aquatic environment.

Toxicological and biochemical response to azinphos-methyl in Cydia pomonella L. (Lepidoptera: Tortricidae) among orchards from the Argentinian Patagonia.

BACKGROUND: Azinphos-methyl is the main insecticide used to control codling moth on apple and pears in Northern Patagonia. The aim of this study was to evaluate the toxicological and biochemical response of diapausing larvae of codling moth in orchards subjected to different insecticide selection pressure. RESULTS: Dose-mortality assays with azinphos-methyl in diapausing larvae of Cydia pomonella L. showed significant differences between the LD(95) from a population collected in one untreated orchard (2.52 microg moth(-1)) compared with that in a laboratory-susceptible population (0.33 microg moth(-1)). Toxicity to azinphos-methyl in field populations of diapausing larvae collected during 2003-2005 was evaluated by topical application of a discriminating dose (2.5 microg moth(-1)) that was obtained from larvae collected in the untreated orchard (field reference strain). Significantly lower mortality (37.71-84.21%) was observed in three out of eight field populations compared with that in the field reference strain. Most of the field populations showed higher esterase activity than that determined in both the laboratory susceptible and the field reference strains. Moreover, there was a high association between esterase activity and mortality (R(2)=0.64) among the field populations. On the other hand, a poor correlation was observed between glutathione S-transferase activity and mortality (R(2)=0.33) among larvae collected from different orchards. CONCLUSIONS: All the field populations evaluated exhibited some degree of azinphos-methyl tolerance in relation to the laboratory susceptible strain. Biochemical results demonstrated that esterases are at least one of the principal mechanisms involved in tolerance to this insecticide.

Pregnancy induced changes in Cox-1, Cox-2 and NOSIII vascular and renal expression.

In order to establish if there is a mutual regulation between COX and NOS in vascular and renal tissue during pregnancy, we measured the protein expression of COX-1, COX-2 and NOSIII by Western blot comparing the thoracic and abdominal aorta and the renal cortex and medulla of non pregnant and pregnant (21st day) Wistar rats. We found there was no difference in the quantity of protein of any of the two isoforms of COX between the two segments of the aorta of non pregnant animals while an increased expression of both COX-1 And COX-2 was found in the abdominal compared to the thoracic segment of the pregnant rats. An increased expression of NOS III was found in the abdominal segment of the aorta form pregnant rats. No changes were found between pregnant and no pregnant animals in the expression of COX-1 and COX-2 in the renal cortex or medulla while an increased expression of NOS III was found in the cortex from pregnant compared to non pregnant animals. These results suggest the influence of pregnancy is not homogeneous along the aorta and also that a balance between prostaglandins and nitric oxide is responsible of the blunted vascular reactivity during pregnancy in the rat.

Pregnancy influence on the vascular interactions between nitric oxide and other endothelium-derived mediators in rat kidney.

Peripheral vascular resistance and sensitivity to circulating pressor and vasoconstrictor agents are blunted during pregnancy. This has been mainly attributed to an increased production of endothelium-derived mediators. The objective of this work was to evaluate if pregnancy changes the relative participation of nitric oxide (NO) and prostaglandins (PG) in respect to the modulation of the increases in renal perfusion pressure induced by phenylephrine (Phe). Dose-response curves were made with gradually increasing doses of Phe using an isolated kidney preparation in the presence of a NO synthase (NOS) inhibitor (L-NAME, 1 microM), a PG-synthesis inhibitor (indomethacin, 1 microM), both, or neither. Also, renal cyclooxygenase (COX-1 and COX-2) and endothelial NOS (eNOS) expression was determined using PCR. The experiments were done in kidneys from nonpregnant and pregnant rats. Our results showed that the relative participation of renal vasoactive mediators seems to change during pregnancy. We found the presence of a COX-1-dependent vasoconstrictor in the middle of pregnancy that was not found in nonpregnant rats. Our results also suggest that there is increased participation of another renal vasodilator substance, the effect of which is observed when NO or PG synthesis is inhibited during late pregnancy. In addition, an apparent interaction between renal eNOS and COX-1 expression was observed: eNOS expression was diminished, while COX-1 was increased during the 2nd week of pregnancy. In contrast, in kidneys from the 3rd week of pregnancy, the expression of these two enzymes was similar.