Type of cell death and the role of acetylcholinesterase activity in neurotoxicity induced by paraoxon in cultured rat hippocampal neurons.

Organophosphate (Ops) neurotoxicity is attributed both to its well-known cholinergic and non-cholinergic effects. In the present study we compared enzymatic and morphologic changes in neurons exposed to paraoxon during one day and one week. The effect of exposure time is important in neurotoxicity of Ops. The longer the exposure time is the more damage is observed in neurons, although there are few investigations about the effect in the post-exposure period. Hippocampal cells were obtained from rat neonates and cultured in Neurobasal/B27. Paraoxon at 50 and 100 microM were added. Inverted microscope and electron microscope were used to study cell morphology and Neutral Red staining was used to measure viability. We also assayed caspase-3 and (acetylcholinesterase) AChE activity. Hoechst staining was utilized to determine the type of cell death. Culture medium was replaced after 24 h in one-day group, however, tests were all carried out at the end of the first week in both group. The results indicate that paraoxon reduced the viability in a dose-dependent manner. Our results do not confirm apoptosis in either group; it seems that the cell death in one-day exposure group was not AChE dependent. In conclusion, present data imply that the toxicity of paraoxon is both dose and duration dependent, which may even remain after the cessation of exposure.

Paraoxon-induced ultrastructural growth changes of rat cultured hippocampal cells in neurobasal/B27.

Organophosphates (OPs) neurotoxicity is attributed both to their well-known cholinergic and recently attended non-cholinergic effects. Since parathion has been observed to be responsible for more cases of poisoning than any other OP insecticides, it is vitally important to investigate other mechanisms, besides cholinesterase inhibition, which can potentially contribute to the neurotoxicity of parathion (or its metabolite, paraoxon). In present study, hippocampal cells obtained from Wistar rat neonates were cultured in neurobasal medium supplemented with B27 serum where different doses of paraoxon were also introduced. The neuronal growth in the control group and those exposed to paraoxon was compared. Phase contrast microscopy, cell staining (Neutral Red) and computer assessment morphometric study (Motic) were used to study cell morphology, viability and type of cell death. Statistical analysis was carried out using one-way ANOVA. There was no clear morphologic differences between neurons in the control group and those exposed to 10 microM paraoxon; however, deformity of the soma was clear in pellets containing higher concentration of paraoxon. Ultrastructure of cells was markedly altered at 50 microM dose of paraoxon as evidenced by gradual discontinuation of cytoplasm, appearing of numerous vacuoles and intracytoplasmic myelin figure. The processes (neurites) did not grow in media containing 100 microM paraoxon or more. Viability decreased with increasing paraoxon especially above 100 microM. In conclusion, the present data reveal that paraoxon, in 30 microM or higher concentrations, induces a decrease in cell growth, followed by cell swelling and neuronal death (possibly necrosis).

Longitudinal study of cyclosporine and lipids in patients undergoing bone marrow transplantation.

The metabolic effects of intravenous cyclosporine on lipids and lipoproteins were studied in 29 allogeneic bone marrow recipients compared with 13 autologous bone marrow patients not requiring cyclosporine therapy. Patients were monitored continuously from 5 days prior to 27 days following transplantation; cyclosporine treatment was initiated 4 days before transplantation. Fasting lipid and lipoprotein levels were measured in serial blood samples throughout the study period. Nutritional supplementation, conditioning regimens and concomitant medications were not significantly different between groups. Furthermore, no significant differences in age, weight, lipid, or lipoprotein levels were found at baseline between the patient groups. Cholesterol, triglyceride, low-density lipoprotein cholesterol, and very low-density lipoprotein cholesterol levels remained unchanged in autologous patients. As compared with baseline values, plasma total cholesterol increased by an average of 26 percent in allogeneic transplantation patients receiving cyclosporine. Similarly, the ratio of low-density lipoprotein to high-density lipoprotein cholesterol was fourfold greater in those patients treated with cyclosporine compared to the autologous group. We conclude that cyclosporine appears to elevate cholesterol levels. Neither acute graft vs host disease nor changes in hepatic function could explain the differences in plasma cholesterol levels between groups.