Thesis (Ph.D.)--Murdoch University, 2004. / Thesis submitted to the Division of Science and Engineering. Includes article published in Annals of Occupational Hygiene, 2001, v. 45 (4), p. 299-308. Bibliography: leaves 267-278.

The endocannabinoid system is composed of endocannabinoids (eCBs), their cognate receptors, and their biosynthetic and catabolic enzymes. Inhibition of serine hydrolases (catabolic enzymes), such as carboxylesterases (CES), might result in the accumulation of eCBs. eCBs, such as 2-arachidonoylglycerol (2-AG), have been shown to increase or reduce inflammation via engagement with cannabinoid receptors on immune cells. This research focuses on exploring the ability of eCBs and their metabolizing enzymes to regulate inflammation. First, a negative feedback mechanism between inflammation and the eCB system was examined by identifying serine hydrolases inhibited by lipopolysaccharide (LPS) stimulation in mice. Ces2g activity was inhibited and Il6 levels were induced in the murine spleen, suggesting a role for this enzyme in an inflammatory response, possibly to limit inflammation. IL-6 did not influence 2-AG hydrolytic activity in human peripheral blood mononuclear cells (PBMCs), but monocytic MAGL was shown to be the predominant 2-AG hydrolytic enzyme in these cells. To investigate a separate mechanism by which serine hydrolases and eCBs may regulate immune responses, mice were treated with chlorpyrifos (CPF), a pesticide known to inhibit serine hydrolases, at doses that do not inhibit acetylcholinesterase in the nervous system. This research is focused on lung tissue since epidemiologic studies have linked pesticide exposures to respiratory diseases. At low doses, Ces1c (adult and neonatal mice) and Ces1d (neonatal mice) were markedly inhibited by CPF (2.5 mg/kg, 7 d, PO). Stimulation with LPS (1.25 mg/kg, IP) following the final CPF dose produced minimal differences in lung immune responses to LPS. In follow up experiments utilizing wild-type and Ces1d-/- mice, a downregulation of Ces1c mRNA in adult Ces1d-/- mice corresponded to an upregulation of Tnfa mRNA in response to LPS in CPF-treated mice. Additionally, Ces1d was found to be expressed in murine alveolar macrophages, suggesting these cells could be used to study the role of CES1 in immunity. Overall, Ces enzymes appear to play a role in immune homeostasis either through a protective mechanism or a negative feedback mechanism to control inflammation.

No description available.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

The thyroid hormone system plays a major role in larval development, growth, and metabolism in fish. Therefore, any anthropogenic alteration in thyroid function could have dramatic effects on individual fitness. In this study Lake Sturgeon, Acipenser fulvescens, larvae were exposed to a commercially used organophosphate pesticide, chlorpyrifos (0, 5, 500 and 2000ng/L), from hatch until the onset of exogenous feeding (~12 days at 14C). The presence of thyroid follicles was first observed at 6 days post hatch (dph). Molecular expression of thyroid receptor α (TRα) increased from 3 to 12dph and then decreased from 12 to 21dph. TRα expression was also significantly higher in brain, liver and muscle at 67dph when compared to TRβ. Of the circulating hormones only free-T3 was consistently measured in larval homogenates from all development time-points sampled. Exposure to chlorpyrifos had no effect on growth or thyroid follicle morphology during the course of the experiment.

Bioremediation was studied in soils containing high concentrations of formulated chlorpyrifos (5 mg kg-1 Dursban® 4E) and atrazine (5 mg kg-1 AAtrex® 4L) using amendments including lignocellulosic sorbents, microbial nutrients (vegetable oil, corn meal and fertilizers), and microbial extracts from organic media previously exposed to these pesticides (chlorpyrifos and atrazine, respectively). Radiolabeled atrazine was used to examine the various dissipation routes in contaminated soil, also amended with lignocellulosic sorbents and microbial nutrients. Both chlorpyrifos and atrazine dissipation from contaminated soils was enhanced by organic-based material amendments. The half-lives of chlorpyrifos based on extractability for soils unamended and amended with vegetable oil and peat moss were 87 and 52 days, respectively. The half-lives of atrazine in unamended and amended soil (vegetable oil, peat moss and fertilizers) were 175 and 40 days, respectively. The leachability of chlorpyrifos from contaminated soil was dramatically reduced by 82% during the first 30 days of incubation in treatments amended with vegetable oil and peat moss while only a 28% of reduction in leachability occurred in the corresponding unamended controls. Only a slight reduction of atrazine leachability was detected in amended treatments after 120 days of incubation. Differences were found in the leachability of chlorpyrifos and atrazine when they were applied to soil either as technical grade or formulated material. The presence of surfactants and other adjuvants in formulated chlorpyrifos (Dursban® 4E) reduced chlorpyrifos leachability in contaminated soil. Chlorpyrifos leachability was reduced by 43% in the formulated chlorpyrifos treatments, whereas there was a negligible decrease in technical chlorpyrifos treated soil during the first 3 days after contamination. Atrazine extractability and leachability was not affected by its formulation (AAtrex® 4L). Amendments with lignocellulosic sorbents and nutrients decreased atrazine®s volatility from contaminated soils. After 16 weeks of incubation, less than 1% of 14C-atrazine was volatilized from incubated soils. Overall, after 16 weeks of incubation less than 4% of 14C-atrazine was mineralized and more radioactivity was recovered from amended treatments than unamended treatments as 14CO2. The major portion of radioactivity (62%) was associated with physisorbed atrazine represented by the ethylacetate extract I from unamended treatments while only 28% of initial applied radioactivity was recovered in the corresponding amended treatments. Based on the sum of radioactivity in humic and fulvic acids, approximately 14% of radioactivity was incorporated or chemisorbed atrazine and its metabolites in both unamended and amended treatments. Forty-five percent of the initially applied radioactivity was associated with alkali insoluble fraction in amended treatments but only 17% of the initially applied radioactivity was detected in the corresponding unamended treatments. Less than 2 % of initial activity associated with physisorbed portions of fulvic acids and alkaline insoluble fraction indicated as the radioactivity in methylene chloride and ethylacetate extract II . Over time, more radioactivity was associated with polar atrazine hydroxylated degradation products. / Ph. D.

Thesis (MTech (Chemical Engineering))--Peninsula Technikon, 2003. / Word processed copy. Summary in English. Includes bibliographical references (leaves 93-96). Also available online.

Obesity is a prevalent disease in which, when compared to individuals of normal weight, obese individuals have noted pathophysiological alterations including increased adiposity and fat mass which may alter toxicokinetics of xenobiotics and therefore alter their toxicities. However, the effects of obesity and the altered pathophysiology accompanying this condition on the toxicity of many widely utilized pesticides have not been established. Organophosphate (OP) pesiticides, including chlorpyrifos (CPS), have historically been widely used for agricultural purposes. CPS and several other notable OPs are no longer registered for household use but still used mainly in agriculture by registered users. CPS is bioactivated to form chlorpyrifos-oxon (CPO) in liver by cytochrome P450 isozymes. Acute toxicity of CPS exposure is inhibition of acetylcholinesterase (AChE) in the central and peripheral nervous systems. The other targets following CPS exposure are the noncholinergic serine hydrolase enzymes: carboxylesterase (CES), fatty acid amide hydrolase (FAAH), and monoacylglycerol lipase (MAGL). Therefore the current study was designed to: 1) determine the time and concentration-dependent effects of CPS on noncholinergic endpoints of OP toxicity utilizing hepatoma cells under normal and steatotic conditions 2) determine if obese phenotype altered the toxicity of CPS, including both cholinergic and non-cholinergic endpoints and 3) determine the effects of high fat diet on CPS bioaccumulation and detoxication by determining CPS and 3,5,6-trichloro-2-pyridinol (TCP) levels at different time points. The current in vitro studies determined that CES is more sensitive to CPS mediated inhibition in normal and steatotic conditions compared to FAAH and MAGL. The in vivo studies determined that CPS (2 mg/kg) exposure produced a greater inhibitory effect on hepatic CES and FAAH in obese animals compared to lean animals which indicates that the obese animals may be at greater risk for CPS mediated alterations in hepatic lipid metabolism upon chronic exposure. Our toxicokinetic studies using a higher dose of CPS (25 mg/kg) did not inhibit AChE in CNS and did not alter the overall CPS levels between normal and obese animals. Peak levels of TCP were decreased in liver of obese animals at 3 hours and overall hepatic TCP levels were significantly decreased in obese animals.

No description available.

Parkinson's disease (PD) is a neurodegenerative Lewy body disorder characterized by severe motor deficits, followed by cognitive dysfunction with progression of the disease. Environmental exposure has been suggested as a possible contributor to the development of PD and this view is linked to the discovery of the nigrostriatal neurotoxin MPTP. MPTP can induce dopamine specific degeneration within the basal ganglia often resulting in motor deficits similar to PD. MPTP used in the C57BL/6 mouse is a widely used animal model of PD. The pyrethroid permethrin (PM), and the organophosphate chlorpyrifos (CPF), can produce changes in dopaminergic nigrostriatal neurons, the primary target of PD and MPTP-induced neurotoxicity. Such insecticide induced changes in the basal ganglia could exacerbate the onset or severity of PD. Chronic exposure to the metal manganese (Mn) can damage the globus pallidus (GP) of the BG, and produce motor deficits similar to PD. Since the GP is part of the BG circuitry essential for motor control, and is synaptically integrated with the nigrostriatal pathway, Mn may exacerbate MPTP-induced neurotoxicity. Because the BG is disynaptically linked to the mesocortical pathway, a dopaminergic pathway that is important for cognition, Mn induced damage in the BG could indirectly affect the mesocortical pathway as well. This study investigated the pesticides, permethrin and chlorpyrifos, and the heavy metal, manganese as possible environmental compounds that could exacerbate PD in the MPTP treated C57BL/6 mouse. The first part of this dissertation used immunohistochemistry to examine insecticide induced effets on MPTP-induced neurotoxicity in the dorsolateral striatum of the C57BL/6 mouse, the principal target of the nigrostriatal pathway. Tyrosine hydroxylase (TH) was used as a marker for loss of dopaminergic neuropil and glial fibrillary acidic protein (GFAP) was used as a marker of glial activation in the striatum. Three experiments assessed effects of 1) PM (200 mg/kg), 2) CPF (50 mg/kg) & 3) PM + CPF, on MPTP (30 mg/kg) neurotoxicity. Immunohistochemistry revealed a decrease in TH staining and an increase in GFAP staining with MPTP (30 mg/kg). A main effect increase in GFAP was observed for PM (200 mg/kg), but not for CPF (50 mg/kg) or PM+CPF. Insecticides, alone or combined, did not alter MPTP-induced toxicity. . However, the absence of the PM-induced increase in GFAP staining following combined insecticide treatment suggests a neuroprotective effect. The next set of experiments in this dissertation looked at the effect of Mn on MPTP-induced neurotoxicity in the nigrostriatal and mesocortical dopaminergic pathways of the C57BL/6 mouse. Inductively Coupled Plasma atomic emission spectrometry revealed striatal Mn levels were significantly increased with multiple dose 100, 50, and 25 mg/kg MnCl2. Administration of Mn (MnCl2 s.c., Days 1, 4, & 7) in the MPTP (20 mg/kg i.p., Day 8) treated C57BL/6 mouse revealed Mn and MPTP interactions for locomotor activity, grip strength, and repeated measures of learning. Mn attenuated the effect of MPTP on striatal DOPAC, and facilitated the effect of MPTP on cortical DA and DOPAC. Mn also attenuated the MPTP induced decrease in cortical DAT. While these data support the notion that insecticides can produce tissue damage in the nigrostriatal pathway, in this case, these insecticide induced changes were not found to be strong enough to facilitate PD-like tissue damage. While Mn did not always facilitate MPTP neurotoxicity in the mesocortical and nigrostriatal dopaminergic pathways, these results demonstrate Mn and MPTP can interact in a complex way to alter dopaminergic function as well as motor and cognitive behavior. Differences in brain uptake mechanisms and metabolism of Mn and MPTP, could explain why combined administration of Mn and MPTP differentially affect dopaminergic activity in the nigrostriatal and mesocortical pathways. / Ph. D.