Remediation of weathered petroleum hydrocarbons, polycyclic aromatic hydrocarbons (PAHs), dioxins, and polychlorinated biphenyls (PCBs) through monitored natural attenuation, in-situ biostimulation, and/or bioaugmentation was assessed using laboratory-scale microcosms. These contaminants of interest (COIs) have persisted in Santa Susana Field Laboratory (SSFL) soils for over 40 years in some cases. The objective of this United States Department of Energy (DOE)-funded study was to determine the potential of the aforementioned remediation methods to reduce COI concentrations in soil and estimate potential biodegradation rates of COIs in SSFL soils. Several types of soil microcosms were established: one set of microcosms was run without amendments to estimate natural attenuation rates at the site; biostimulation was tested by addition of nitrogen and phosphorus, rice hulls, and biosurfactant (soya lecithin), another set was augmented with the white-rot fungus Phanerochaete chrysosporium, and gamma-irradiated microcosms served as sterilized controls. Soil samples were collected and analyzed for dioxins, PCBs, PAHs, and extractable fuel hydrocarbons (EFH) after 0, 4, and 8 months of incubation. Soil contamination in the microcosms initially consisted of primarily heavily chlorinated dioxins and PCBs, longer petroleum hydrocarbons (21-40 equivalent carbon chain length), and PAHs with 4-6 aromatic rings. Small decreases in PAH, PCB, and dioxin soil concentrations were observed, but these decreases were not statistically significant. EFH concentrations were inflated at the final sampling event, but they appeared to reduce for two of three soils (Soils A and C) tested at the second sampling event. No COI concentration reductions were statistically significantly during 8 months of incubation. Because petroleum hydrocarbons were primarily longer-chain hydrocarbons in the C21 to C40 EFH range, it is likely that lighter hydrocarbons had been preferentially degraded, leaving the more recalcitrant longer-chain hydrocarbons in the soil. Dioxin concentrations appeared to decrease in some cases, but these reductions were not statistically significant at the 95% confidence level. Larger PAHs (4-6 rings) comprise the majority of residual PAH soil contamination. Given that concentrations of these PAHs have not decreased significantly during this 8-month long study, it is likely that these larger PAH contaminants are somewhat recalcitrant and will take a long time to biodegrade. Similarly, little or no PCB biodegradation was observed which is not surprising because the PCBs are heavily chlorinated, and bacterial biodegradation of these highly chlorinated compounds is reported to occur only under anaerobic conditions. The primary dioxin congener present in soils was octachlorodibenzodioxin (OCDD), which is the heaviest-chlorinated dioxin congener. Like PCBs, this compound requires anaerobic conditions for reductive dechlorination, and these are not present at the site. Total dioxin concentrations decreased in the microcosms amended with Phanerochaete chrysosporium, although this decrease was not statistically significant due to variability of dioxin concentrations measured in the soil. No decrease in tetrachlorodibenzodioxin toxicity equivalence was observed with P. chrysosporium bioaugmentation, and this parameter is important in terms of dioxin toxicity. Soil vapor analyses performed at the site indicate highly aerobic soil conditions. To mimic site conditions as closely as possible, experimental microcosms were maintained incubated in aerobic conditions. Although fungi have been reported to degrade PCBs and dioxins under aerobic conditions, the microcosms augmented with Phanerochaete chrysosporium did not show statistically significant biodegradation of PCBs. Contaminant sequestration in the soil may also have contributed to the lack of observed biodegradation because the COIs at this site are highly weathered. However, even microcosms augmented with a surfactant (soya lecithin), which would be expected to solubilize sequestered COIs, did not show significant biodegradation.

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Nitrated polycyclic aromatic hydrocarbons (nitro-PAH) are environmental contaminants that have been identified in extracts of particulates obtained from automobile exhaust, diesel exhaust, and power plant emissions. One of the most abundant nitro-PAH, 1-nitropyrene, has been found to be a powerful bacterial mutagen and a carcinogen in animal tests. In a bacterial strain (Salmonella tvphimurium TA98) 1- nitropyrene undergoes reduction and concomitant covalent binding to DNA, presumably via the N-hydroxy-l-aminopyrene. This labile compound was prepared by the ascorbic acid reduction of 1-nitrosopyrene and was characterized by high performance liquid chromatography (HPLC), combined liquid chromatography-mass spectrometry (LC-MS), ultraviolet-visible (UV-VIS), fluorescence and nuclear magnetic resonance (NMR) spectroscopy (1H and 15N) . In addition the pH stability and some reactions of this compound were studied. The 1,6- and 1,8-hydroxylaminonitropyrenes derived from the very potent mutagens 1,6- and 1,8 dinitropyrene were also synthesized and characterized by high performance liquid chromatography, ultraviolet-visible and nuclear magnetic resonance spectroscopy (1H and 15N) . / Thesis / Master of Science (MSc)

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Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

Yes / Epigallocatechin gallate (EGCG), the main ingredient in green tea, holds promise as a potential treatment for pulmonary arterial hypertension (PAH). However, EGCG has many drawbacks, including stability issues, low bioavailability, and a short half-life. Therefore, the purpose of this research was to develop and optimize an inhalable EGCG nano-liposome formulation aiming to overcome EGCG’s drawbacks by applying a design of experiments strategy. The aerodynamic behaviour of the optimum formulation was determined using the next-generation impactor (NGI), and its effects on the TGF-β pathway were determined using a cell-based reporter assay. The newly formulated inhalable EGCG liposome had an average liposome size of 105 nm, a polydispersity index (PDI) of 0.18, a zeta potential of −25.5 mV, an encapsulation efficiency of 90.5%, and a PDI after one month of 0.19. These results are in complete agreement with the predicted values of the model. Its aerodynamic properties were as follows: the mass median aerodynamic diameter (MMAD) was 4.41 µm, the fine particle fraction (FPF) was 53.46%, and the percentage of particles equal to or less than 3 µm was 34.3%. This demonstrates that the novel EGCG liposome has all the properties required to be inhalable, and it is expected to be deposited deeply in the lung. The TGFβ pathway is activated in PAH lungs, and the optimum EGCG nano-liposome inhibits TGFβ signalling in cell-based studies and thus holds promise as a potential treatment for PAH. / This study was supported by Schlumberger Foundation’s faculty for the future in the form of PhD funding awarded to Fatma Haddad.