Les hydrocarbures aromatiques polycycliques (HAPs), tels que le benzo(a)pyrène (B[a]P), sont des contaminants environnementaux ubiquistes générés lors de la combustion de matière organique. Ces composés ont été associés au développement d'effets toxiques sur la santé humaine, notamment des effets cancérigènes et immunotoxiques, principalement liés à l'activation du récepteur aux hydrocarbures aromatiques (RAh). Parmi les cellules du système immunitaire, les lymphocytes T apparaissent comme des cibles majeures des HAPs. Des résultats antérieurs, obtenus au laboratoire, ont montré que l'activation des lymphocytes T humains en culture primaire conduit à l’augmentation de l'expression et de la fonction du RAh, suggérant la capacité accrue de ces cellules à répondre à une exposition aux HAPs. Nos objectifs sont : (1) de déterminer les effets du B[a]P sur les profils d'expression génique dans les lymphocytes humains normaux en utilisant des approches à haut débit telle que l'analyse transcriptomique sur puce à ADN, (2) d’évaluer les effets génotoxiques et immunotoxiques du B[a]P en mesurant respectivement les dommages à l'ADN induits et leurs actions immunosuppressives et (3) d’analyser la modulation de ces effets en présence d'autres HAPs. Notre travail identifie les lymphocytes T humains normaux comme un bon modèle pour étudier les effets génotoxiques et immunotoxiques des HAPs, et pour prédire les problèmes de santé humaine liés à l’exposition à ces contaminants. Il permet également de mieux comprendre la régulation par les HAPs de la réponse immune et propose de nouveaux biomarqueurs potentiels de l'exposition à ces contaminants environnementaux. / Polycyclic aromatic hydrocarbons (PAHs), such as benzo(a)pyrene (B[a]P), are ubiquitous environmental contaminants generated during organic matter combustion. These compounds have been associated with the development of toxic effects on human health, including carcinogenic and immunotoxic effects, mainly related to Aryl hydrocarbon Receptor (AhR) activation. Among the immune system cells, T lymphocytes appear as major targets of PAHs. Previous results, obtained in the laboratory, have shown that activation of primary human T lymphocytes leads to a functional AhR expression increase, suggesting their ability to respond to PAH exposure. Our specific aims are: (1) to determine the effects of B[a]P on gene expression profiles in human normal lymphocytes by using large-scale approaches such as microarray-based transcriptome analysis, (2) to monitor the genotoxic and immunotoxic effects of B[a]P by measuring DNA damage and immunosuppressive actions, respectively and, (3) to analyze the modulation of these effects by the presence of other PAHs. Our work propose primary cultures of activated human T lymphocytes as a good model for studying both genotoxic and immunotoxic effects of environmental contaminants such as PAHs and predicting human health issues. It also gains a comprehensive insight into the immune response regulation after PAH exposure and provides potential new biomarkers of exposure to these environmental contaminants.

Polycyclic aromatic hydrocarbons, produced by anthropological and natural activities, are hazardous through formation of oxidative radicals and DNA adducts. Growth of Mycobacterium sp. strain KMS, isolated from a contaminated soil, on the model hydrocarbon pyrene induced specific proteins. My work extends the study of isolate KMS to the gene level to understand the pathways and regulation of pyrene utilization. Genes encoding pyrene-induced proteins were clustered on a 72 kb section on the KMS chromosome but some also were duplicated on plasmids. Skewed GC content and presence of integrase and transposase genes suggested horizontal transfer of pyrene-degrading gene islands that also were found with high conservation in five other pyrene-degrading Mycobacterium isolates. Transcript analysis found both plasmid and chromosomal genes were induced by pyrene. These processes may enhance the survival of KMS in hydrocarbon-contaminated soils when other carbon sources are limited. KMS also grew on benzoate, confirming the functionality of an operon containing genes distinct from those in other benzoate-degrading bacteria. Growth on benzoate but not on pyrene induced a gene, benA, encoding a benzoate dioxygenase α-subunit, but not the pyrene-induced nidA encoding a pyrene dioxygenase α-subunit; the differential induction correlated with differences in promoter sequences. Diauxic growth occurred when pyrene cultures were amended with benzoate or acetate, succinate, or fructose, and paralleled delayed expression of nidA. Single phase growth and normal expression of benA was observed for benzoate single and mixed cultures. The nidA promoters had potential cAMP-CRP binding sites, suggesting that cAMP could be involved in carbon repression of pyrene metabolism. Growth on benzoate and pyrene requires gluconeogenesis. Intermediary metabolism in isolate KMS involves expression from genes encoding a novel malate:quinone oxidoreductase and glyoxylate shunt enzymes. Generation of C3 structures involves transcription of genes encoding malic enzyme, phosphoenolpyruvate carboxykinase, and phosphoenolpyruvate synthase. Carbon source modified the transcription patterns for these genes. My findings are the first to show duplication of pyrene-degrading genes on the chromosome and plasmids in Mycobacterium isolates and expression from a unique benzoate-degrading operon. I clarified the routes for intermediary metabolism leading to gluconeogenesis and established a potential role for cAMP-mediated catabolite repression of pyrene utilization.

No description available.

Polycyclic aromatic hydrocarbons are environmental chemicals that are produced during incomplete combustion of coal, oil, gas, and garbage. Toxic effects of these compounds are mediated via the ligand activated Aryl Hydrocarbon Receptor (AHR) signaling pathway. To enable the study of the AHR signaling mechanism, our lab has generated many human proteins using recombinant DNA technology. This thesis documents the design and synthesis of a number of proteins of the AHR deletion construct CΔ553. The bacterial expressed and purified fusion proteins could be utilized as antigen to generate antibodies and be used for cellular delivery. The purified protein was immunogenic in rabbits and produced significant amount of polyclonal antibodies. In western blot analysis, the antibodies were able to the detect baculovirus expressed AHR and different recombinant proteins of the AHR. The polyclonal antibodies were also used in the gel-shift assay to show the AHR dependent gel shift. Cellular delivery CΔ553 was achieved using the protein transduction domain from the HIV-1 virus transactivating protein (TAT). In order to deliver the CΔ553 into mammalian cells, an expression vector was constructed to generate the TAT-CΔ553 fusion protein. The TAT-CΔ553 fusion protein was successfully transduced into two mammalian cells-HeLa and HepG2. The in vivo function of TAT-CΔ553 was determined using the luciferase reporter plasmid assay. The transduced protein was functional; it competed with the AHR and heterodimerize with ARNT in both HeLa and HepG2 cells at a concentration of 3.8x103 nM and 18 nM respectively. Since there an apparent similarity between the basic region of TAT-PTD and CΔ553, we examined the transduction potential of CΔ553. Western blot analysis indicated that the extent of denatured protein transduction was comparable for CΔ553 and TAT-CΔ553 in HepG2 cells. Thus CΔ553 might have intrinsic transduction capability.

Crude oil commonly enters freshwater aquatic ecosystems as thin sheens forming on the water surface. Oil contains mixtures of toxic compounds called polycyclic aromatic hydrocarbons (PAHs), some of which are known to be photodynamic, increasing toxicity when combined with ultraviolet radiation. Benthic macroinvertebrate communities are commonly utilized as bioindicators, and as such rely on abundant data in literature concerning benthic macroinvertebrates' relative tolerances to a wide range of pollutants. A series of 10 plastic traps, half of which were filtered from UV radiation, were deployed in an urban pond for 27 days to determine colonization preferences of benthic macroinvertebrates to UV exposure. Results of this in situ experiment indicated that the majority of aquatic insects collected from traps inhabited the UV exposed treatment group, particularly the nonbiting midge, Chironomidae. A series of bioassays were then completed to investigate the sensitivities of a Chironomidae species to thin sheens of crude oil in the presence and absence of UV radiation. All bioassays were conducted using 10 day old Chironomus dilutus larvae cultured in the lab. The series of C. dilutus bioassays were all conducted under the same water quality parameters, temperatures, and oil sheen dosing methods, under a 16:8 photoperiod and exposed to 16 h UV per day. Five replicates (n = 20) were loaded into 350 mL glass crystallizing dishes and exposed to four treatments for 96 hours: no UV/with sheen, with UV/no sheen, both UV and oil sheen, and a control. Three assays with 175 mL water volume were completed, one with no sediment, one with silica sand, and one with fine sand. Sediment type had a significant effect on mortality (p < 0.0001), but significant effects of UV or PAHs were not found. Two more C. dilutus assays were completed with identical parameters as the latter two with sediment, except a decreased water volume of 90 mL was used to bring the sheen closer to sediment. There was a significant effect of PAHs on mortality (p = 0.0003), however, no clear trend showing PAHs driving mortality. Results showed no significant effects of UV, sediment type, or relationship between UV and PAHs. A final bioassay was completed to compare phototoxic effects of PAHs on pelagic organisms with benthic organisms. This bioassay used test species Daphnia magna, for a 48 h exposure period with five replicates (n = 10), 8 h UV exposure and 16:8 photoperiod. All individuals in the control group and UV only group exhibited 0% mortality, and with sheen/with UV treatment group resulted in 100% mortality following the 48 h exposure. The results of the D. magna exposure showed significant effects of PAHs (p < 0.0001), UV (p = 0.037), and photoinduced toxicity (p = 0.024), and were consistent with similar bioassays in the literature. This study suggests that C. dilutus are at low risk to photo-induced oil sheen toxicity.

Structurally precision graphene nanoribbons (GNRs) have attracted great interest considering their prospective applications as organic carbon materials for nanoelectronics. The electronic properties of GNRs not only critically depend on the edge structure and width but also on the heteroatom type, doping position, and concentration. Motivated by the recent undisputable progress in the synthesis of stable boron-doped polycyclic aromatic hydrocarbons (B-PAHs), considerable efforts have been devoted to the precision synthesis of the corresponding boron-doped GNRs (B-GNRs) via bottom-up synthesis approach in recent years in view of the extraordinary ability of boron doping on modulating their physiochemical properties. In this review, an overview of the bottom-up organic synthesis of B-GNRs, including the precursor design and synthesis, structure characterization of the resulting B-GNRs, and investigation of their electronic properties is provided. Moreover, the future challenges and perspectives regarding the bottom-up synthesis of B-GNRs are also discussed. The authors hope that this review will further stimulate the synthesis and device integrations of B-GNRs with a combined effort from different disciplines.

No description available.

No description available.

Polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) are common contaminants of sediment, soils and biological tissues. These compounds pose a significant risk to biological and ecosystem health and functioning due to these compounds being mutagenic, carcinogenic and are known to disrupt the endocrine system. The bioaccumulation and biomagnification potential that these compounds possess mean that they are capable of affecting the entire food chain and are not limited to the organisms that are directly in contact with the compounds Even though there has been an increase in the attention on identifying the presence and impacts that these compounds may have in South Africa, the level of attention is lower than what it is in other countries around the world. Although South Africa has guidelines in place for other pollutants, such as metals, there are no such guidelines in place to monitor PAHs and PCBs.. Industries are known to release both PAHs and PCBs, mainly from incomplete burning processes and the release of oils and fuels in the case of PAHs, and from heat transfer fluids in the case of PCBs. Durban Bay and surrounding areas of eThekwini, KwaZulu-Natal are highly industrialised with many aquatic systems, in which these contaminants are likely to deposit. The aim of this research was to determine the concentrations of these compounds by means of chemical analysis and additionally biological analysis, using the H4IIE-luc bioassay and compare these levels to international guideline levels. It was found that the concentrations of the 23 analysed PAHs were 6.5–3 235.6 ng.g-1 and the concentration of the PCBs analysed were 0–113.83 ng.g-1. Many sites were found to be in exceedance of the guideline limits, particularly in the harbour. Toxic equivalency factors (TEF) were used to gauge the toxic equivalency (TEQ) of the PAHs and PCBs that were found. The TEQs were generally low, and were below any guideline levels. The assay revealed the extract containing the PCBs had a bioassay equivalence (BEQ) of 0–93.54 pgTCDDeq. g-1 and the extract containing the PAHs of 0–776.08 pgTCDD-eq.g-1. With a proportion of the sites exceeding guideline limits. The BEQ results were two to three orders of magnitude greater than the TEQs calculated from the concentrations determined by the instrumental analysis, however, followed a similar trend. Additionally chemical analysis was not performed on a full suite of compounds that are able to elicit a response from the cells, which could be a reason why the BEQ and TEQ did not follow a similar trend among some of the samples. It would have been more beneficial to have performed chemical analysis on the 16 priority PAHs (as determined by the United States Environmental Protection Agency), the dioxin-like PCBs and polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDFs) which are all capable of eliciting a response from the cells and have TEF values. The areas that were most affected by contamination of these compounds was the harbour and surrounding canals, and there was point source contamination along the Umhlatuzana, Umbilo, and Amanzimnyana Rivers. All dl-PCBs should be chemically analysed at all the sample areas, as these cause adverse effects to biota. In addition to this, biota should be sampled to determine concentrations of the compounds to determine bioavailability and the degree of bioaccumulation in the food chain. Utilising biomarkers it would be possible to determine stresses of fish. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014

Polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) are common contaminants of sediment, soils and biological tissues. These compounds pose a significant risk to biological and ecosystem health and functioning due to these compounds being mutagenic, carcinogenic and are known to disrupt the endocrine system. The bioaccumulation and biomagnification potential that these compounds possess mean that they are capable of affecting the entire food chain and are not limited to the organisms that are directly in contact with the compounds Even though there has been an increase in the attention on identifying the presence and impacts that these compounds may have in South Africa, the level of attention is lower than what it is in other countries around the world. Although South Africa has guidelines in place for other pollutants, such as metals, there are no such guidelines in place to monitor PAHs and PCBs.. Industries are known to release both PAHs and PCBs, mainly from incomplete burning processes and the release of oils and fuels in the case of PAHs, and from heat transfer fluids in the case of PCBs. Durban Bay and surrounding areas of eThekwini, KwaZulu-Natal are highly industrialised with many aquatic systems, in which these contaminants are likely to deposit. The aim of this research was to determine the concentrations of these compounds by means of chemical analysis and additionally biological analysis, using the H4IIE-luc bioassay and compare these levels to international guideline levels. It was found that the concentrations of the 23 analysed PAHs were 6.5–3 235.6 ng.g-1 and the concentration of the PCBs analysed were 0–113.83 ng.g-1. Many sites were found to be in exceedance of the guideline limits, particularly in the harbour. Toxic equivalency factors (TEF) were used to gauge the toxic equivalency (TEQ) of the PAHs and PCBs that were found. The TEQs were generally low, and were below any guideline levels. The assay revealed the extract containing the PCBs had a bioassay equivalence (BEQ) of 0–93.54 pgTCDDeq. g-1 and the extract containing the PAHs of 0–776.08 pgTCDD-eq.g-1. With a proportion of the sites exceeding guideline limits. The BEQ results were two to three orders of magnitude greater than the TEQs calculated from the concentrations determined by the instrumental analysis, however, followed a similar trend. Additionally chemical analysis was not performed on a full suite of compounds that are able to elicit a response from the cells, which could be a reason why the BEQ and TEQ did not follow a similar trend among some of the samples. It would have been more beneficial to have performed chemical analysis on the 16 priority PAHs (as determined by the United States Environmental Protection Agency), the dioxin-like PCBs and polychlorinated dibenzo-p-dioxins (PCDD) and polychlorinated dibenzofurans (PCDFs) which are all capable of eliciting a response from the cells and have TEF values. The areas that were most affected by contamination of these compounds was the harbour and surrounding canals, and there was point source contamination along the Umhlatuzana, Umbilo, and Amanzimnyana Rivers. All dl-PCBs should be chemically analysed at all the sample areas, as these cause adverse effects to biota. In addition to this, biota should be sampled to determine concentrations of the compounds to determine bioavailability and the degree of bioaccumulation in the food chain. Utilising biomarkers it would be possible to determine stresses of fish. / MSc (Environmental Sciences), North-West University, Potchefstroom Campus, 2014