To date, most studies on the fate and removal of endocrine disrupting compounds (EDCs) and pharmaceutical and personal care products (PPCPs) in wastewater focus on their fate in municipal wastewater treatment plants, and mostly under aerobic condition. There are limited studies related to anaerobic condition and (to our knowledge) no study on the removal of EDCs in landfill leachate by AnMBR. Moreover, for most studies under anaerobic condition, the removal of EDCs was only reported in the liquid phase; solid phase extraction was not reported, thereby preventing mass balance in the studies. This research was conducted to investigate the potential of AnMBR for reduction of organic strength and removal of EDCs in landfill leachate. A novel lab-scale upflow anaerobic sludge blanket (UASB) reactor equipped with dual-flat sheet ultrafiltration and microfiltration membrane modules was designed and constructed to test the potential to remove EDCs and traditional landfill leachate constituents (COD, turbidity). The target EDC was 17β-estradiol (E2), a prevalent female hormone used for contraceptives and hormone replacement therapy. Due to the nature of packaging and widespread use in households, the entry of E2 into landfills is highly likely, and has been reported. The quantification of E2 from liquid phase in this project is performed by the use of solid-phase microextraction (SPME) with GC/MS. Batch assays were conducted to determine the anaerobic biodegradability of E2 as well as to measure the respective distribution coefficients of E2 to PAC, colloids and anaerobic sludge biomass. In the adsorption batch assays, it was found that the PAC has stronger adsorption potential than anaerobic sludge. The adsorption potential of E2, E1 and EE2 on sludge follows the order E2>EE2>E1 which correlates to the Kow values (4.01, 3.67, 3.1, respectively). However, all three compounds showed the same adsorption potential to the Norit 20B PAC. The biodegradability of E2 was investigated in both liquid and solid phase and under several conditions such as methanogenesis, methanogenesis with aid from PAC, and methanogenesis with additional alternative electron acceptors added (sulfate and nitrate). E2 was found to transform to E1 under all tested conditions. The compounds are present in both liquid and solid phase. E2 and E1 were not detected (< 4ng/L and <10ng/L, respectively) in the liquid phase after 25 days in most cases except the case of adding additional sulfate. The AnMBR was designed, fabricated and operated for 2 years. During the stable condition period of the AnMBR, the high removal efficiencies of COD and E2 achieved were around 92% and 98%, respectively. However, E2 was still detected in the effluent at average concentrations of 30-40 µg/L range. To expand hormone retention and removal by the AnMBR, as well as to control membrane fouling, powder activated carbon (PAC) was added to the reactor. After the PAC was added, the concentration of E2 was reduced to less than the detection limit (4ng/L) in both MF and UF effluents. The log removal of E2 in the AnMBR system increased immediately from 1.7 without PAC to 5.2 after PAC was added. This study demonstrated that the AnMBR has high potential for removal of E2, and with aid from PAC, the AnMBR can remove E2 from landfill leachate to levels below detection limit.

L'utilisation massive des retardateurs de flamme organohalogénés dans les produits manufacturés se solde, à ce jour, par une dispersion importante de ces composés dans l'environnement et par leur accumulation au sein des espèces aviaires notamment. À cet effet, il fut mesuré les plus fortes concentrations du congénère BDE-209 (composante principale du mélange Déca-BDE) couplé à une forte détection des isomères du Déchlorane Plus (syn- et anti-DP) dans les tissus de goéland à bec cerclé (GBC) nichant dans le fleuve Saint-Laurent, en aval de Montréal. En raison de leur forte charge en halogène et leur poids moléculaire important, le BDE-209 et le DP ont été considérés comme étant peu biodisponibles et très stables d'un point de vue physicochimique. Or, en ce qui a trait au BDE-209, plusieurs produits de débromination ont également été quantifiés dans ces mêmes tissus, mais leurs sources restent encore ambiguës. Par ce travail de recherche, nous avons donc tenté de vérifier l'hypothèse proposant que le BDE-209 et les isomères du DP représentent de bons substrats à l'action des enzymes de biotransformation isolées des microsomes hépatiques de GBC, résultant ainsi en une série de produits déhalogénés. Cette étude a donc pour but d'élucider le potentiel de biotransformation du BDE-209 et des deux isomères du DP à l'aide d'un essai de biotransformation in vitro (90 min) utilisant des microsomes hépatiques de GBC naturellement exposés, et ce, en les comparant à un congénère métabolisable (BDE-15) et à un autre étant résistant à la dégradation enzymatique (BDE-153). Basé sur un gradient d'activité EROD hépatique (faible, moyen et élevé), les oiseaux ayant une activité EROD moyenne ont démontré une diminution significative d'environ 20% du BDE-15 alors qu'aucune dégradation significative n'a été obtenue pour le BDE-153, -209 et pour les isomères du DP. Également, le pourcentage de dégradation du BDE-15 entre les groupes d'activité EROD « faible » et « moyen » a varié d'un peu plus de 10% laissant entrevoir qu'une certaine induction enzymatique serait favorable aux fins de la biotransformation de ce congénère. En somme, le présent travail ne permet pas d'expliquer la présence des produits de débromination du BDE-209 dans les tissus du GBC étant donné l'absence de biotransformation observée lors de l'essai in vitro. Ces produits pourraient toutefois avoir été absorbés par leur alimentation et/ou leur environnement quoique l'on ne puisse exclure que des processus métaboliques affectant l'intégrité moléculaire du BDE-209 aient véritablement lieu chez le GBC. Quant aux isomères du DP, il semble qu'ils ne soient pas susceptibles à la biotransformation dans le cadre du design expérimental réalisé. ______________________________________________________________________________ MOTS-CLÉS DE L’AUTEUR : Retardateur de flamme halogéné, polybromodiphényléther (PBDE), Déchlorane Plus (DP), oiseau, biotransformation in vitro, microsomes hépatiques

Phytosterols are naturally occurring compounds which regulate membrane fluidity and serve as hormone precursors in plants. They also have the potential to cause endocrine disturbances in aquatic animals at concentrations as low as 10 µg/L. Wastewaters from several industries which process plant matter can contain phytosterols at concentrations in excess of the above-stated level. Despite their endocrine disruption potential, very little is known about phytosterol physical properties and their biotransformation potential in biological treatment systems. Aerated stabilization basins (ASBs) are common biological treatment systems in North American pulp and paper mills. ASBs are large open lagoons which use tapered surface aeration to remove COD and prevent sulfate reduction in the water column. Phytosterols are released from wood during the pulping process and a small fraction enters the wastewater stream during washing of the pulp. Therefore, phytosterols may be exposed to aerobic or anaerobic environments depending on their solubility and solid-liquid partitioning behavior. The overall objective of this research was to systematically and quantitatively assess the biotransformation potential of phytosterols in biological treatment systems and to examine conditions leading to reduction of these compounds in wastewater effluent streams. The results of this research showed that phytosterols are sparingly soluble with aqueous solubility below 1 µg/L when present as a mixture. Phytosterols have a strong affinity to adsorb to solids and dissolved organic matter. The affinity for aerobic biomass was greater than for wastewater solids. The stigmasterol desorption rate and extent from wastewater solids increased with an increase in pH from 5 or 7 to 9. Phytosterols were biotransformed under aerobic conditions but not under sulfate-reducing or methanogenic conditions by stock cultures developed in this study. Biotransformation under nitrate-reducing conditions could not be confirmed conclusively. The continuous-flow system was successful in removing 72 to 96% of phytosterols. Biotransformation accounted for 23, 14 and 41 % of campesterol, stigmasterol and β-sitosterol removal, respectively. Phytosterols accumulated in the reactor sediment and accounted for 97 % of the total phytosterols remaining in the system. Phytosterols can be removed from wastewater streams during biological treatment by a combination of biotransformation and solids partitioning and control of system pH, DO and available carbon and energy sources can increase the degree of phytosterols removal. The results of this research can be used to engineer effective biological treatment systems for the removal of phytosterols from pulp mill wastewaters and other phytosterol-bearing wastewater streams.

In this study, the ability of various chemicals to be biotransformed into mutagens by plant cells was investigated. Two thiocarbamate herbicides, diallate and triallate, the sulfonylurea herbicide chlorsulfuron, and the aniline derivative m-phenylenediamine were tested for their ability to revert Salmonella typhimurium (strains TA100 and TA98) in the presence and absence of Nicotiana tabacum (TX1) cell cultures in liquid suspension. Chlorsulfuron and m-phenylenediamine were also tested in the presence and absence of Brassica napus cv. 'Topas' cells. Diallate was found to be activated by TX1 cells into a mutagen that induces base-pair substitution mutations. In the presence of the TX1 plant cell line, chlorsulfuron significantly increased the number of mutations on the strain TA98 of Salmonella. Tobacco TX1 cells did not activate triallate into a mutagen. m-Phenylenediamine was activated into a mutagen by TX1 and Brassica cells as detected by Salmonella TA98. This aniline derivative, in the absence of plant cells and at concentrations higher than 20 $ mu$ Moles/plate, was also able to significantly increase the number of TA98 revertants as compared to the control plants. Finally, Brassica napus cells activated chlorsulfuron into a mutagen that induces frameshift mutations.

This dissertation combines laboratory and field experiments to investigate the mechanisms of atropisomer enrichment for chiral polychlorinated biphenyls (PCBs) and their metabolites in organisms. Stereoselective biotransformation and bioaccumulation were identified as two major reasons for the different environmental fate of PCB atropisomers. Other affecting factors, such as presence of nanoparticles and changes in feeding ecology of organisms, also affect the fate of chiral contaminants. In vitro incubations of rat cytochrome P-450 2B1 (CYP2B1) isozyme with chiral PCBs indicated that different biotransformation kinetics and competition among PCB congeners or between atropisomers were two main factors affecting atropisomer enrichment. Different interactions between chiral PCB congeners or atropisomers with rat CYP2B1 may occur at the molecular level. Non-racemic meta-hydroxylated-PCBs (5-OH-PCBs) were the major metabolites. CYP-mediated stereoselective formation of dihydroxylated PCBs from OH-PCBs was observed. Gold nanoparticles affected biotransformation activity of rat CYP2B1 and changed PCB atropisomeric composition, directly by electrostatic interaction, or indirectly by changes to the surrounding ionic strength. Thus, stereoselective metabolism of chiral PCBs and OH-PCBs by CYPs is a major mechanism for atropisomer enrichment of PCBs and their metabolites in the environment, with the degree of enrichment dependent, at least in part, on charged nanoparticles and stereoselective interference of atropisomers with each other at the enzyme level. The atropisomer compositions of chiral PCBs were measured in the marine biota of Cumberland Sound (Canada) and Svalbard (Norway). High trophic level organisms, including harp seal, beluga, and narwhal reported for the first time, had species-specific atropisomer signatures, likely due to a combination of in vivo biotransformation and trophic transfer. PCB chiral signatures in Greenland sharks supported the hypothesis that some of these PCB atropisomer compositions shifted over time and space, possibly due to a change in feeding ecology. To our knowledge, this is the first report to investigate temporal trends of PCB atropisomer signatures in Arctic biota.

Thesis (Ph. D.)--University of Oklahoma. / Bibliography: leaves 234-253.

Ulm, Univ., Diss., 1999. / http://vts.uni-ulm.de/query/longview.meta.asp?documentid=356.

Thesis (M.S.)--University of Missouri-Columbia, 2005. / "December 2005" The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Vita. Includes bibliographical references.

Os estudos da biotransformação têm-se mostrado um método eficiente para reações que envolvam a obtenção de compostos com interesse comercial. Dentre estes produtos utilizados podemos destacar monoterpenos por constituírem uma variedade de compostos com grande aplicação tanto no âmbito farmacêutico, como em perfumes, cosméticos e alimentos. As suspensões de células vegetais e os fungos filamentosos têm sido largamente usados em reações de biotransformação destes metabólitos secundários como substratos exógenos. A habilidade das culturas celulares e fungos em transformar tais compostos vêm sendo dirigidos principalmente para a obtenção de derivados oxigenados de maior valor agregado, principalmente visando à produção de flavorizantes, aromatizantes e fragrâncias. As suspensões de células vegetais apresentam sistemas enzimáticos mais complexos, podendo levar a produtos específicos, enquanto os microrganismos, por serem mais simples e de crescimento mais rápido, geralmente promovem mais rapidamente a metabolização dos substratos.Neste contexto, o presente trabalho descreve a investigação do potencial de bioconversão dos monoterpenos (R)-(+)-α-pineno, (S)-(-)-α-pineno e (S)-(-)-β- pineno, bem como do óleo de terebentina, para avaliação de suas potencialidades frente a reações de biotransformação mediadas pelos microrganismos Cunninghamella echinulata ATCC 9245, Mortierella isabellina NRRL 1757 e Cunninghamella elegans ATCC 36112 e por suspensões de células vegetais de Catharanthus roseus. Todos os sistemas estudados mostraram habilidade na bioconversão dos substratos testados. / Biotransformation has been an efficient method to obtain important commercial compounds such as monoterpenes, that have a range of applications in the pharmaceutical, perfumery, cosmetics and food industries. Plant cell cultures and filamentous fungi suspensions have been widely used in biotransformation reactions of monoterpenes as exogenous substrates. The ability of the cells and fungi cultures to transform these compounds has being mainly used for the synthesis of oxygenated derivatives aiming at the production of flavours and fragrances. The plant cell cultures suspensions have complex enzymatic systems with the production of specific compounds, where as filamentous fungi, which are structurally simpler and fast growing, can promote quicker transformations reactions. In this context, the present work describes the evaluation of bioconversion of the monoterpenes (R)-(+)-α-pinene, (S)-(-)-α-pinene, (S)-(-)-β-pinene, as well as turpentine oil using the filamentous fungi suspensions of Cunninghamella echinulata ATCC 9245, Cunninghamella elegans ATCC 36112 and Mortierella isabellina NRRL 1757, aswer as Catharanthus roseus plant cell cultures. All evaluated systems were capable of bioconverting the studied substrates. The main product obtained from pinenes and turpentine oil was α-terpineol.

Charles University Faculty of Pharmacy in Hradec Králové Department of Pharmacology & Toxicology Student: Anna Giannitsi Supervisor: RNDr. Jakub Hofman, Ph.D Title of diploma thesis: The role of biotransformation enzymes in the resistance of cancer cells against standard cytostatics Drug resistance is currently one of the major problems of chemotherapy. Tumor cells are able to defend themselves against the effect of cytostatic drugs due to various mechanisms which leads to a failure of anticancer therapy. The effort to describe new mechanisms of resistance and to develop new therapeutic methods, which would limit this therapeutic obstacle, is logically the subject of many studies. The activity of drug metabolizing enzymes and the subsequent decrease of intercellular concentration of anticancer drugs belongs to one of the possible mechanisms of pharmacokinetic resistance. Enzymes of I. and II. phase of biotransformation participate in this phenomenon. Cytochromes P450, main enzymes of the I. phase, play a major role in the metabolism of many cytostatic agents producing either pharmacologically active or inactive metabolites. Increased expression in tumors and the involvement of individual isoforms into the overall metabolism of cytostatic, which is deactivated by their activity, seems to be one of the...