Les rejets d'effluents liquides des Centres Nucléaires de Production d'Electricité (CNPE) sont constitués d'un mélange de substances stables et radioactives. L'exposition d'organismes à des substances en mélange peut faire l'objet d'interactions diverses, conduisant à une augmentation ou à une diminution des effets observés. Afin d'identifier de possibles interactions dans le cas de mélanges de substances caractéristiques (en termes de toxicité et de quantité) des rejets de CNPE, l'effet d'un mélange binaire composé de cuivre et d'HTO a été étudié sur le modèle d'algue unicellulaire Chlamydomonas reinhardtii. Dans un premier temps, la toxicité de l'HTO a été analysée. L'HTO s'est révélée être peu toxique envers notre modèle biologique. L'effet le plus sensible et le plus précoce est une augmentation du stress oxydant (dès 40 kBq mL-1 - 0.13 µGy h-1). Lors de l'exposition des cellules algales au mélange HTO/Cu, une interaction a été révélée au niveau du stress oxydant cellulaire, celui-ci étant supérieur à une simple addition de l'effet des deux substances. Cette interaction peut s'expliquer par une augmentation de l'internalisation du cuivre en présence d'HTO, mais aussi potentiellement par des interactions toxiques directes (notamment sur les processus de régulation du stress oxydatif). En conclusion, il a été démontré que l'effet de substances stables et radioactives en mélange peut être supérieur à l'addition. Bien qu'étant uniquement représentative du mélange binaire HTO/Cu, cette étude montre néanmoins de potentielles interactions entre substances stables et radioactives, à considérer lors de l'évaluation des risques écologiques relatifs aux rejets de CNPE. / Liquid releases by Nuclear Power Plants (NPP) are composed of a mixture of radioactive and non-radioactive substances. When organisms are exposed to mixtures of contaminants the resultant toxicity can be enhanced, or reduced, due to interactions. In order to identify potential interactions between substances released by NPP, two substances representative of such effluents (in term of toxicity and of quantity) were selected for studies: Tritiated water (HTO) and copper (Cu). Effects of this binary mixture were studied on the unicellular green algae Chlamydomonas reinhardtii. HTO, when examined along, was not very toxic to C. reinhardtii. The most sensitive and early effect of HTO was an increase in oxidative stress at concentrations of 40 kBq mL-1 (0.13 µGy h-1). Algae exposure to the binary mixture HTO/Cu induced interactive effects on oxidative stress. Reactive Oxygen Species production was higher from exposure to the mixture of contaminants than the addition of the effect from each substance individually. This interaction was explained by an enhanced copper uptake by the alge when in the presence of HTO. The observed supra-additive effect could also be due to direct toxic interactions, especially on the antioxidant system. To conclude, this study showed that the effects of a mixture of radioactive and non-radioactive substances can be greater than what would be predicted based on mere addition of individual effects. Even thought this binary mixture is just a small part of NPP effluents, the study showed that potential interactions should be considered when determining ecological risks too aquatic ecosystems from NPP effluents.

My MSc thesis is focused on parametric designs of Phase I clinical trials, using the continual reassessment method. A parametric model with unknown parameters is assumed. The observations are either toxic or nontoxic. Observations of toxicities are used to update the posterior distribution. Dose selection for the next patient is based on the estimated toxicity probability. The objective is to identify the maximum tolerated dose to be used in Phase II clinical trials. We introduce a new class of parametric functions for the continual reassessment method. This class is formed with the cumulative distribution function of the normal distribution. The major advantage is that we can choose different normal distributions to model different toxicity probability functions. We conduct simulation studies and compare our new design with the existing parametric designs, and have found that our design performs better by choosing the appropriate values of the mean and variance. / October 2016

Beyond the expected DBP exposure from drinking water, dermal from bathing, and inhalation, occurrence in food and beverage items can provide an additional occurrence pathway. Synthetic dyes are often added to beverages for aesthetic purposes and many are in the form of a reactive azo (-N=N-) dye or triarylmethane dye, both with a characteristic aromatic ring. The presence of dyes in beverages that are reconstituted with disinfected tap water pose the risk of reactions with the residual chlorine in the drinking water resulting in decolorization, and of greater concern, disinfection byproduct formation. Additionally, oral over-the-counter (OTC) medications contain chemical constituents that when reconstituted with tap water present a risk of DBP formation. Several studies were performed to evaluate the kinetic decay rates of the dyes and drugs in disinfectants, and the effects of water quality conditions on DBP formation. Commercial beverage products and OTC medications were evaluated for the DBPs that were detected in the free chlorine-treated precursor samples. The dye and drugs precursors followed second order kinetics, with the fastest rates for brilliant blue and phenylephrine in chlorinated water. The effects of water properties on precursor degradation and DBP formation was complex due to the influence of characteristics of precursor molecules. The cytotoxic and anti-estrogenic responses were measured in the dye and drug precursors and their respective beverages and OTC medications, to determine potential links. Mio Energy showed estrogenic character and Alka Seltzer induced an anti-estrogenic and cytotoxic response, however there were no clear linkages between the beverage/ medication and their respective dye and drug precursors.

During recent years most of the successful developments in pyrethroids have been primarily concerned with structural or compositional variations. As a part of our continuing interest in pyrethroid insecticides, nitrogen-containing compounds having pyrethroid-like structures were synthesized. Seven prolinate compounds, N-(substituted)-phenyl-prolinates and N-carbobenzoxy-prolinates were coupled with known pyrethroid alcohols. These structural variations which "locked in" a specific conformation between the nitrogen and chiral a-carbon in the acid moiety of fluvalinate were studied to determine the influence of certain conformations on insecticidal toxicity. The toxicity data for the prolinate compounds showed intermediate mortality against nonresistant cockroaches. It was concluded that the conformation imposed by the proline ring portion of the esters was probably close to the favored conformation for interaction of fluvalinate-like pyrethroids with the insect receptor site. A second series of nitrogen-containing compounds, twenty-five carbamate esters resulting from the condensation of N-isopropyl-(substituted)-anilines and N-alkyl-(substituted)-benzylamines with appropriate pyrethroid alcohols were studied for insecticidal activity. These studies were conducted on pyrethroid-susceptible houseflies. Some of the carbamate esters exhibited high toxicity when synergized by piperonyl butoxide. For example, the toxicity ( LD 50 ) of O-a-cyano-3-phenoxyfaenzyl-N-a,a-dimethyl-4-bromo-benzyl carbamate was 0.012 ug/g, which is significantly greater than that reported for the potent pyrethroid, fenvalerate. Correlations of insecticidal activity with respect to structure and conformational factors of the carbamate esters have been made. The N-isopropyl substituent decreases insecticidal activity in the N-benzyl-derived compounds, while the N-isopropyl substituent enhances activity in the N-phenyl-derived compounds. Certain substituents on the phenyl ring of both analogs greatly affect insecticidal potency of the carbamate esters. Also, some alkyl substituents (especially, a,cx-dimethyl and a-cyclopropyl groups) on the benzylic carbon of the benzylamine series enhance toxicity. The a,a-dimethyl branching of the N-benzyl carbamate approximates the steric shape given by the gemdimethyl group for conventional cyclopropane ring-containing pyrethroids. The N-benzyl compounds are significantly synergized by piperonyl butoxide, particularly those in which the carbamate nitrogen atom is mono-substituted.

The biological activities of some homeosterically related analogs of dihydrofolic acid have been examined involving pyrimido[4,5-b][l,4]oxazine (8-oxapteridine) derivatives. It is anticipated that these compounds might interfere with the essential intermediary metabolic functions of the vitamin and thus serve as potential chemotherapeutic agents. Preliminary toxicity studies in microbial assay systems were disappointing; however, inhibitory effects were demonstrated in cell free enzyme systems. A comparison of the structure/activity relationships was determined using two folic acid coenzyme systems, dihydrofolate reductase and thymidylate synthetase. The 2-amino-4-hydroxy-6-(substituted)-8-oxapteridines were generally more effective inhibitors than the corresponding 2,4-diamino analogs. The relative biological activity of a series of 2-amino-4-hydroxy-6-ω-phenylalkyl derivatives were examined, and the most active derivative was the 6-phenylethyl analog which appears to function as a mixed-type inhibitor involving partially competitive and partially non-competitive inhibition.

Lead is a central nervous system poison. Healthy People 2010 established a target blood lead level (BLL) for children of 0 μg/dL by 2010, but is silent with regard to any changes in BLLs standards for working age adults. In this paper, the relation of BLL to performance on two neurobehavioral tests was assessed in working age adults (N = 4909; Age 20 to 59 years; 51.4% Female) employing data from the Third National Health and Nutrition Survey (NHANES 111). Multiple linear regression analyses indicated a significant effect of BLL on time taken to complete an attention demanding cognitive task (Symbol Digit Substitution Task, SDST) but not accuracy of performance of the SDST or simple reaction time, after controlling for confounding variables of age, sex, race-ethnicity, and education. Persons with BLL ≥5 μg /dL took longer (multivariate adjusted mean = 23.6 Sec, SE = 0.30) compared to individuals with BLLs <5 μg /dL (mean = 22.5 Sec, SE = 0.14). The results suggest that lead burden in working age persons impairs central nervous processes involving executive mental functions (decision speed and attention). The findings, if confirmed by case control and or cohort studies would indicate a need to reconsider currently accepted lead levels in working age adults.

This study describes the development of a novel method of three dimensional perfused cell culture for in vitro toxicity testing. Multiple parallel perfused microbioreactors (TissueFlex^TM) were adopted to provide a well-controlled cell culture environment. Alginate and collagen type I, commonly used as hydrogel scaffolds to support cell culture, were tested as the scaffolding materials for this application. Alginate supports cell proliferation, but does not support cell attachment. Collagen gel (type I), good for cell attachment but with poor mechanical strength, could be used at the high concentration of 5mg/ml to prevent the degradation of the gel. Improvement of collagen biomechanical property by a purpose-designed compressor to physically induce cross-linking showed promising results and merits further study. The suitability of alamarBlue&reg; assay, a common non-toxic non-destructive viability assay method, was confirmed for this study and the protocol was optimised. To demonstrate the effectiveness of three dimensional perfused cell culture, human mesenchymal stem cells (MSC) seeded in collagen type I were employed to test the cell inhibition of two antibiotics, trimethoprim and pyrimethamine. The results displayed the perfusion system has greater advantage and sensitivity than the static system, as does these of 3D scaffolds, compared with 2D. Such differences are related to the continuous supply of fresh culture medium to keep cells at a stable pH, temperature, oxygen, and a more physiological like environment. The cytotoxicity of two stereoisomer compounds, obtained confidentially from Pfizer. Ltd., was assessed using the developed method and compared to conventional 2D static and perfused culture by using rat adipose mesenchymal stem cells. The results successfully distinguished toxic and non-toxic compounds and also demonstrated that the 3D perfused system improved the prediction of drug toxicity over 2D culture. 3D perfused bioreactors were applied to hepatotoxicity study using freshly isolated rat hepatocytes. Only algimatrix^TM supported hepatocyte spheroid formation among those tested including collagen type I, alginate beads, poly lactic acid fibres, and Algimatrix^TM. A new variation of TissueFlex^TM bioreactor with micro-patterned surface, designed specifically for hepatocyte self-assembly culture without use of any scaffold, was tested. The results demonstrated that, compared with the standard sandwich culture, the self-assembly culture in the micro-patterned bioreactors showed high cell viability, biomarkers expression, as well as more physiological immunocytochemistry. Moreover, the differential gene expression indicated that self-assembly culture could provide more relevant information regarding metabolising processes than the 2D sandwich culture, which would potentially improve hepatotoxicity prediction. In conclusion, 3D perfused cell culture for in vitro toxicity testing improved the predictivity, reliability and physiological relevance of drug toxicity compared to traditional 2D culture.

<p>Following the phase out of polybrominated diphenyl ethers (PBDEs), manufacturers turned to several alternative flame retardants (FRs) to meet flammability standards. Organophosphate FRs (OPFRs), and in particular tris (1,3-dichloropropyl) phosphate (TDCPP), have been increasingly detected in textiles and foam padding used in a variety of consumer products including camping equipment, upholstered furniture, and baby products. Like PBDEs, OPFRs are additive, meaning that they are not chemically bound to the treated material and can more readily leach out into the surrounding environment. Indeed, OPFRs have been detected in numerous environmental and biological matrices, often at concentrations similar to or exceeding that of PBDEs.</p><p>Although OPFRs have been in use for several decades, relatively little is known regarding their potential for adverse human and environmental health consequences. However, based on their structural similarity to OP pesticides, they may have analogous mechanisms of toxicity. OP pesticide toxicity is classically associated with cholinesterase inhibition, resulting in cholinergic intoxication syndrome. OPFRs have been shown to be ineffective cholinesterase inhibitors, however chlorpyrifos (CPF) and other OP pesticides have been shown to elicit adverse effects on developing organisms through other mechanisms.</p><p>The main objective of this research project was to evaluate the toxicity of four structurally similar OPFRs (TDCPP; tris (2,3-dibromopropyl) phosphate, (TDBPP); tris (1-chloropropyl) phosphate (TCPP) and tris (2-chloroethyl) phosphate (TCEP)) in comparison to chlorpyrifos (CPF), a well-studied OP pesticide. A combination of in vitro and in vivo models was used to elucidate potential mechanisms as well as functional consequences of exposure in developing organisms. </p><p>In the first research aim, a series of in vitro experiments with neurotypic PC12 cells was used to evaluate the effects of four structurally similar OPFRs (TDCPP, TDBPP, TCEP, or TCPP) and CPF on neurodevelopment. The effects of TDCPP were also compared to that of BDE-47, a major component of the commercial PentaBDE mixture. In general, TDCPP elicited similar or greater effects when compared to an equimolar concentration of CPF. All OPFRs tested produced similar decrements in cell number and altered phenotypic differentiation, while BDE-47 had no effect on cell number, cell growth, or neurite growth. </p><p>For the second research aim, zebrafish (Danio rerio) were used to evaluate the effects of the same suite of chemicals on early development. TDCPP, TDBPP, and CPF elicited overt toxicity (e.g., malformations or death) within the concentration range tested (0.033-100 µM). TDBPP was the most potent with 100% mortality by 6 days post fertilization (dpf) at &#8805;3.3 µM. CPF and TDCPP showed equivalent toxicity with malformations observed in at 10 µM and significant mortality (&#8805;75%) at &#8805;33 µM. There was no overt toxicity among TCEP- and TCPP-exposed fish. All test chemicals affected larval swimming behavior on 6 dpf at concentrations below the overt toxicity threshold. Parent chemical was detected in all in embryonic (1 dpf) and larval (5 dpf) tissues. TDCPP and TDBPP showed rapid and extensive metabolism.</p><p>Finally, for the third aim, juvenile (45-55 dpf) zebrafish were exposed to CPF (1 µg/g food) or TDCPP (Low TDCPP = 1 µg/g food; High TDCPP = 40 µg/g food) via diet for 28 days followed by a 7 day depuration period where all treatments received clean food. A dietary exposure was chosen to more closely recapitulate exposure in humans. Samples were collected at seven time points throughout the experiment: days 0, 7, 14, 21, 28, 30, 35. Whole tissues were collected for tissue accumulation and histopathology endpoints. Viscera and brain were dissected and flash frozen separately for DNA damage analyses. </p><p>Tissue measurements of CPF, TDCPP, and the metabolite bis (1,3-dichloropropyl) phosphate (BDCPP) were often below the method detection limit, however when present there was a trend towards increased accumulation with treatment and time. On Day 7 Low TDCPP caused a dramatic but transient increase in DNA damage in both viscera and brain that returned to control levels by Day 14. Similar results have been seen previously with other genotoxicants and may be due to CPF and High TDCPP inducing an adaptive response prior to the 7 day sampling point. All treatments shifted the neurohypophysis to adenohypophysis ratio (NH/AH; Day 7 only) and significantly increased thyroid follicle activation (Day 14). Finally High TDCPP affected gonad maturation, causing a significant increase in ovary follicle development (Day 14) and a transient but marked decrease in testes maturity (Day 7). Taken together these data suggest that dietary exposure to TDCPP and CPF elicits DNA damage in brain and viscera and alters endocrine function in juvenile zebrafish. Importantly, analyses were restricted to the first three time points (Days 0, 7, and 14) due to the emergence a disease among the experimental colony. Although these samples were collected prior to the disease becoming apparent, it remains a potential confounder of the current results.</p> / Dissertation

Introduction : La présence des moisissures et de leurs mycotoxines dans les matrices complexes de papiers est peu étudiée. Notre travail a porté sur la recherche de mycotoxines sur les papiers patrimoniaux altérés par le foxing et les papiers peints moisis issus de logements dont les habitants ont été diagnostiqués comme porteurs de symptômes allergiques et du syndrome des bâtiments malsains. Objectifs : Identifier les espèces fongiques de ces deux types de supports papiers et y déterminer la production de métabolites fongiques. Matériels et Méthodes : Le foxing a été caractérisé par des techniques pluridisciplinaires (physiques, biologiques, bioanalytiques, tests de cytotoxicité). Les métabolites fongiques dans les extraits hydro-organiques de ces papiers ont été recherchés et identifiés par spectrométrie de masse afin d'évaluer s'ils pouvaient être reliés aux espèces fongiques détectées par microbiologie. Puis le risque toxique de ces extraits de papiers a été évalué sur un modèle cellulaire in vitro. Résultats : Pour le foxing, nous avons pu en exclure une origine métallique et montrer que les micro-organismes présents dans celui-ci sont essentiellement des espèces fongiques. Pour les papiers peints, des pics relatifs à des métabolites fongiques ont été retrouvés. Grâce à l'ensemencement individuel d'espèces fongiques sur papier peint et à l'aide de la MS2, nous avons pu relier les composés de m/z 401 et 487 à S. chartarum. Les tests de cytoxicité ont montré une augmentation significative de la cytotoxicité des cellules A549 avec certains papiers peints moisis par rapport au papier peint témoin. Les cellules A549 ont montré une surexpression du TNF-α, de l'IL-8 et du CYP 1A1, après contact avec ces mêmes papiers peints. Discussion-Conclusion : Nous n'avons détecté aucune mycotoxine dans le foxing excluant ainsi d'éventuels liens entre inhalation de mycotoxines émanant de vieux manuscrits et symptomatologie respiratoire. Pour les habitants des logements et leurs symptômes respiratoires, il est difficile de les relier à une espèce fongique donnée ou à un métabolite donné, bien que S. chartarum ait pu être mis en cause. Ces premiers résultats doivent être confirmés par des études ultérieures / Introduction: Little study has been carried out on the presence of fungi and their mycotoxins in complex paper matrices. Our work has focused on finding mycotoxins on heritage paper altered by the foxing and wallpapers from moldy homes whose residents have been diagnosed with symptoms of allergies and sick building syndrome. Objectives: To identify the fungal species of these two types of paper and to determine the production of fungal metabolites. Materials and methods: The foxing has been characterized using multi-disciplinary technics (physical, biological, bioanalytical, cytotoxicity assays). The fungal metabolites in the hydro-organic extracts of these papers were sought and identified by mass spectrometry to assess whether they could be related to fungal species detected by microbiology. The toxicological risk of wallpaper extracts was then evaluated on model in vitro cells. Results: For the foxing, we could exclude a metal origine and we showed that the microorganisms present are mainly fungal species. For wallpapers, fungal metabolites were found. By seeding individual fungal species on wallpaper and using MS2, we were able to link the compounds of m/z 401 and 487 to S. chartarum. Cytotoxicity tests showed a significant increase in cytotoxicity of A549 cells with moldy wallpaper in comparison with the control wallpaper. A549 cells showed an overexpression of TNF-α after contact with these wallpapers as well as a significant overexpression of IL-8 and CYP 1A1.Discussion-Conclusion: We detected no mycotoxin in foxing, thus excluding possible links between inhalation of mycotoxins from old manuscripts and respiratory symptoms. For people in their homes and respiratory symptoms, it is difficult to draw any relationships to a given fungal species or a metabolite although S. chartarum has been implicated. These initial results should be confirmed by further study

Aluminium toxicity is the main limiting factor in crop production on acid soils. The main symptom of aluminium toxicity is a rapid inhibition of root growth, but the mechanism of root growth cessation remains unclear. In this diploma thesis we deal with the question of whether phospholipases PLDα1 and PLDδ may play a role in the mechanism of aluminium toxicity. We compared the responses of plants lacking PLDα and PLDδ with WT plants. Growth analysis of roots was performed in hydroponic conditions. The most sensitive part of roots was transient zone in which cells were dying earlier. It was further found that pldα1 plants were less sensitive on aluminium toxicity because their roots showed less growth inhibition than WT. Pldδ plants did not differ from WT plants in their response to aluminum. During further analysis of the pldα1 reactions, it was found that the root cells were capable of cell expansion during aluminum toxicity, and the cellular malformations were formed on the roots. This phenomenon was associated with faster reorientation and even depolymerization of cortical microtubules in response to toxic aluminium in pldα plants compared to WT plants. The results indicated that PLDα1 molecule affects the stability of cortical microtubules. Microtubules were less stable and they depolymerized...