Metal contaminated stormwater, i.e. surface runoff in urban areas, can be treated in percolation systems, ponds, or wetlands to prevent the release of metals into receiving waters. Plants in such systems can, for example, attenuate water flow, bind sediment, and directly accumulate metals. By these actions plants affect metal mobility. This study aimed to examine the accumulation of Zn, Cu, Cd, and Pb in roots and shoots of plant species common in stormwater areas. Furthermore, submersed plants were used to examine the fate of metals: uptake, translocation, and leakage. Factors known to influence metal accumulation, such as metal ion competition, water salinity, and temperature, were also examined. The following plant species were collected in the field: terrestrial plants – Impatiens parviflora, Filipendula ulmaria, and Urtica dioica; emergent plants – Alisma-plantago aquatica, Juncus effusus, Lythrum salicaria, Sagittaria sagittifolia, and Phalaris arundinacea; free-floating plants – Lemna gibba and Lemna minor; and submersed plants – Elodea canadensis and Potamogeton natans. Furthermore, the two submersed plants, E. canadensis and P. natans, were used in climate chamber experiments to study the fate of the metals in the plant–water system. Emergent and terrestrial plant species accumulated high concentrations of metals in their roots under natural conditions but much less so in their shoots, and the accumulation increased further with increased external concentration. The submersed and free-floating species accumulated high levels of metals in both their roots and shoots. Metals accumulated in the shoots of E. canadensis and P. natans derived mostly from direct metal uptake from the water column. The accumulation of Zn, Cu, Cd, and Pb in submersed species was in general high, the highest concentrations being measured in the roots, followed by the leaves and stems, E. canadensis having higher accumulation capacity than P. natans. In E. canadensis the Cd uptake was passive, and the accumulation in dead plants exceeded the of living with time. The capacity to quickly accumulate Cd in the apoplast decreased with successive treatments. Some of the Cd accumulated was readily available for leakage. In P. natans, the presence of mixtures of metal ions, common in stormwater, did not alter the accumulation of the individual metals compared to when presented separately. It is therefore, proposed that the site of uptake is specific for each metal ion. In addition cell wall-bound fraction increased with increasing external concentration. Further, decreasing the temperature from 20ºC to 5ºC and increasing the salinity from 0‰ to 5‰ S reduced Zn and Cd uptake by a factor of two. In P. natans the metals were not translocated within the plant, while in E. canadensis Cd moved between roots and shoots. Thus, E. canadensis as opposed to P. natans may increase the dispersion of metals from sediment via acropetal translocation. The low basipetal translocation implies that neither E. canadensis nor P. natans will directly mediate the immobilisation of metal to the sediment via translocation. To conclude, emergent and terrestrial plant species seem to enhance metal stabilization in the soil/sediment. The submersed plants, when present, slightly increase the retention of metals via shoot accumulation.

This thesis is concerned with the synthesis and evaluation of regenerable chalcogen containing antioxidants. Variously substituted 2,3-dihydrobenzo[b]selenophene-5-ol antioxidants were evaluated in order to gain information about structure/reactivity-relationships. Within the series explored, the most regenerable unsubstituted compound inhibited lipid peroxidation for more than 320 minutes when assayed in a two-phase lipid peroxidation model in the presence of N-acetylcysteine (NAC). α-Tocopherol which could inhibit lipid peroxidation for 90 minutes under similar conditions was therefore easily outperformed. The antioxidant activity of the parent was also documented in an aqueous environment. The best catalyst quenched/inhibited ROS production by neutrophils and PMA-stimulated macrophages more efficiently than Trolox. In addition, over a period of seven days, no disruption in proliferation for the cell lines used was observed when exposed to our synthetic compound or Trolox at a concentration of 60 µM. 3-Pyridinols substituted with alkyltelluro groups in the ortho-position were more regenerable in the two-phase model than their corresponding para-substituted analogues in the presence of NAC and also inhibited autoxidation of styrene in a catalytic fashion in homogenous phase in the presence of N-tert-butoxycarbonyl cysteine methyl ester (LipCys), a lipid-soluble analogue of NAC. The best inhibitors quenched peroxyl radicals more efficiently than α-tocopherol. They could also catalyze reduction of organic hydroperoxides in the presence of thiols and therefore mimic the action of the glutathione peroxidase enzymes. Mechanisms for the catalysis are proposed. Octylthio, octylseleno and octyltelluro analogues of butylated hydroxyanisole (BHA) were synthesized and evaluated. Among these, the tellurium compound was superior to α-tocopherol in the presence of NAC both when it comes to quenching capacity and regenerability.  Organochalcogen substituent effects in phenolic compounds were studied by using EPR, IR and computational methods.

Due to a policy of no release, oil sands process-affected water (OSPW) produced by the surface-mining oil sands industry in North Eastern Alberta, Canada, is stored on-site in tailings ponds. There is concern regarding the toxic effects of OSPW on aquatic organisms. Knowledge of the chemical composition and toxicity of OSPW is limited. Research is necessary for potential remediation and release of OSPW back into the environment. Due to the large volume and persistency of OSPW, active efforts are necessary for the remediation of OSPW before release and habitat reclamation. Currently, ozonation is considered one possible method for remediation of OSPW by reducing the concentrations of dissolved organic compounds, including naphthenic acids (NAs), which are considered among the primary toxic constituents. However, further work is needed to evaluate the effectiveness of ozonation in reducing the toxicity of OSPW and to ensure that ozonation does not increase the toxicity of OSPW. The overall objective of this work was to determine the toxic effects of OSPW on endocrine disruption and embryo development, using both in vitro and in vivo models, and the effectiveness of ozone treatment for reducing the toxicity of OSPW. In the first study, untreated and ozone-treated OSPW were examined for effects on sex steroid production using the H295R cell line steroidogenesis Assay. The results indicate that exposure to untreated OSPW can significantly decrease synthesis of testosterone (T) and increase synthesis of 17β-estradiol (E2) by 0.55±0.06 and 2.0±0.13-fold, respectively, compared to that of control groups (ρ < 0.05). These effects were due to increased aromatase enzyme activity and decreased E2 metabolism. The results also suggest that ozonation is an effective treatment to reduce concentrations of NAs in OSPW without altering steroidogenesis. In the second study, the T47D-kbluc (estrogen responsive) and MDA-kb2 (androgen responsive) cell assays were used to determine whether OSPW might act as either agonists or antagonists of the estrogen receptor (ER) or androgen receptor (AR), respectively. The estrogenic responses to untreated OSPW were significantly greater by 2.6±0.22-fold compared to control group (ρ < 0.05). Exposure to untreated OSPW produced significant antiandrogenic response in the presence of 0.01, 0.05 and 0.1 nM T by 16±6.5%, 47±7.6% and 75±9.7%, respectively, of that of the corresponding concentrations of T alone (ρ < 0.05). The results suggest that compounds in the dissolved organic fraction of OSPW have estrogenic and anti-androgenic properties, acting as ER agonists and/or AR antagonists. Ozonation of the OSPW partially mitigated the antiandrogenicity but had no effect on the estrogenicity of OSPW. In the third study, the endocrine-disrupting effects of OSPW and ozone-treated OSPW were determined by quantifying relative changes in the abundances of transcripts of genes along the brain-gonad-liver (BGL) axis in male and female fathead minnows (Pimephales promelas). The results indicate that OSPW has endocrine-disrupting effects at all levels of BGL axis and these effects of impaired expression of genes along the BGL axis are sex specific. For example, exposure to OSPW resulted in significantly greater abundances of transcripts of vtg (Vitellogenin), chg-l (Choriogenin L) and chg-h (Choriogenin H minor) by 4.9±1.2, 5.4±1.5 and 3.4±0.78-fold, respectively, compared to those of control groups (ρ < 0.05) in livers from male fathead minnow. However, in livers from female fathead minnows, exposure to OSPW resulted in significantly lesser abundances of transcripts of vtg, chg-l and chg-h by 0.002±0.0011, 0.022±0.007 and 0.036±0.024-fold, respectively, compared to those of control fish (ρ < 0.05). Ozonation of OSPW attenuated the effects on abundances of transcripts of some genes, and the attenuation was more prominent in males than in females. However, impact of ozonation on endocrine-disrupting effects of OSPW was less evident than in the in vitro studies described in Chapter 2 and 3. The results also provide a mechanistic basis for the endocrine-disrupting effects of OSPW from other studies, including impaired reproduction of fathead minnows exposed to OSPW. In the final study the effects of untreated, ozone-treated, and activated charcoal-treated OSPW (OSPW, O3-OSPW, and AC-OSPW) on the survival, growth, and development of embryos of fathead minnows were determined. Compared to the control group, which had an embryo survival rate of 98±2.1%, survival was significantly less when exposed to OSPW (44±7.1%; ρ < 0.05). Eggs exposed to untreated OSPW exhibited a significantly greater rate of premature hatching, and embryos exhibited more frequent spontaneous movements. Incidences of hemorrhage (50±3.4%), pericardial edema (56±7.1%), and malformation of the spine (38±5.4%) were significantly greater in embryos exposed to OSPW compared to control group (ρ < 0.05). Significantly greater concentrations of ROS (1.7±0.11-fold), and greater abundances of transcripts cyp3a, gst, sod, casp9, and apopen (2.4±0.34, 2.2±0.26, 3.1±0.74, 3.3±0.57 and 2.4±0.25-fold, respectively) compared to control groups (ρ < 0.05), indicated that exposure to OSPW caused oxidative stress, which can result in damage to mitochondria and promote activation of caspase enzymes and apoptotic cell death. Removal of dissolved organic constituents in OSPW by ozone treatment, or by activated charcoal, significantly attenuated all of the adverse effects associated with untreated OSPW. The results suggest that the organic fraction of OSPW can negatively impact the development of fathead minnow embryos through oxidative stress and apoptosis, and that ozonation attenuates this developmental toxicity. Overall, the findings from the research described in this thesis provide novel and important insights into the toxicity and mechanisms of the toxicity of OSPW with respect to endocrine disruption and development of embryos of fish. In addition, the research provides compelling evidence that ozonation might be an effective method for accelerating the remediation of OSPW. The results of the research might help regulators develop effective strategies for reclamation, remediation and potential release of OSPW back to the environment.

Incorporation of nanoparticles (NPs) into consumer products is on the rise and human exposure to NPs is unavoidable. Currently, there is insufficient data to assess the safety of nanoparticles. I conducted a series of five studies using the zebrafish model to determine which NP components (i.e., core material or surface functionalization) contribute to biological responses and how ionic strength influences these results. The first study employed a systematic, rapid embryonic zebrafish assay to identify specific responses to precisely engineered lead sulfide (PbS-NPs) and gold nanoparticles (AuNPs) functionalized with different surface ligands. Lead sulfide nanoparticles functionalized with either 3-mercaptopropanesulfane (MT) or sodium 2,3-dimercaptopropanesulfonate (DT) ligands with nearly identical core sizes caused differential responses at the same concentration. I determined that the different responses were because MT-functionalized NPs released more soluble lead ions than DT-functionalized NPs due to different decomposition and oxidation rates. The second study investigated the different biological responses of three NPs identified during toxicity screening of a gold nanoparticle library. AuNPs functionalized with 2-mercaptoethanesulfonic acid (MES), N,N,N-trimethylammoniumethanethiol (TMAT), or 2-(2-(2-mercaptoethoxy)ethoxy)ethanol (MEEE), induced differential biological responses in embryonic zebrafish at the same concentration. Exposure to MES-AuNPs induced sublethal effects, while TMAT-AuNPs were embryo-lethal and MEEE-AuNPs were benign. Gold tissue concentration was confirmed to be similar in exposed embryos using inductively coupled-mass spectrometry. Microarrays were used to gain insight to the causes of the different responses. This approach identified that MES- and TMAT-AuNPs perturbed inflammatory and immune responses. These differential biological responses may be due to misregulated transport mechanisms causing numerous downstream defects unique to each surface functional group‟s property. In the next study, I tested the long-term consequences of developmental exposure to TMAT-, MES, and MEEE-AuNPs, and showed that MES- and TMAT-AuNPs affected larval behavior that persisted into adulthood. During the course of these investigations, I found that high ion concentration in exposure solutions results in NP agglomeration, presenting a problem for NP testing in the zebrafish model. For the fourth study, I focused on solving this by determining that zebrafish can be raised in nearly ion-free media without adverse consequences. When 3-MPA-AuNPs were dispersed in this new low ionic media, I observed adverse responses in the embryonic zebrafish toxicity assay, but not when the NPs were suspended in high ionic media. Thus, I demonstrated that the media greatly influences both agglomeration rates and biological responses, but most importantly, that the zebrafish is insensitive to external ions. The fifth study focused on the adverse response observed when embryonic zebrafish were exposed to 3-MPA-AuNPs. Exposed larvae failed to respond to a touch in the caudal fin at 120 hours post fertilization (hpf). Addition of a neuromuscular stimulus, nicotine, revealed the exposed embryos were not paralyzed, but experienced a reduction in axonal projections. A global genomic analysis (RNA-seq) using embryos exposed to 3-MPA-AuNP and MEEE-AuNPs (non-toxic control) from 6 to 120 hpf suggested that neurophysiological and signal transduction processes were perturbed. Functional analysis of the data led to the hypothesis that the most elevated gene, early growth response 1 (EGR-1), impacts axonogenesis in the caudal fin, interfering with glutaminergic synapses and preventing the connection of sensory neurons and touch perception. Although MEEE-AuNPs did not cause morphological defects, the RNA-seq analysis identified that these NPs perturbed immune and inflammatory system processes. Collectively, these results suggest that surface functional groups drive the differential responses to nanomaterials. The five studies summarized here confirm that a systems toxicological approach using the zebrafish model enables the rapid identification of structure-activity relationships, which will facilitate the design of safer nano-containing products. / Graduation date: 2013

Metal toxicity is implicated in neurotoxicity, nephrotoxicity, aging and cancer. Protein oxidation resulting from oxidative stress is now known to be involved in metal toxicity. However, proteomic responses to metal induced oxidative stress have not been characterized. By using the yeast as a model, we characterized these changes occurring in response to sub-lethal doses of metals. Several proteins involved in protein synthesis, ribosome assembly decreased while antioxidant defenses, proteins involved in sulfur metabolism, and glutathione synthesis and ubiquitin increased following metal exposure. We also show that metals induced temporal and targeted protein oxidation independent of protein abundance. Among the targets were glycolytic enzymes and heat-shock proteins. As a consequence, glycolytic enzyme activities decreased whereas the levels and activities of the enzymes of the alternative pathway for glucose metabolism, pentose phosphate pathway (PPP) increased. True to prediction, we also found increased flow through the PPP as measured by elevated levels of NADPH and glutathione. NADPH and glutathione are crucial for maintaining the redox balance in the cell. Thus, rerouting of glucose metabolism into PPP is considered to be beneficial to the organism. Among the oxidation targets is a glycolytic protein, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) that is required for apoptosis in neuronal cells. We show that not only is GAPDH required for metal induced apoptosis in yeast but also the levels of GAPDH transcript and protein increase in the cytosol and the nucleus in an isoform specific fashion. Such changes strongly implicate the role of GAPDH in yeast apoptosis. This work provides evidence for the involvement of targeted protein oxidation in metal toxicity, shows the overlaps and differences in the mechanism of copper and cadmium toxicity, allows comprehension of how metabolic processes respond to metal stress and explores the potential of GAPDH as a sensor of oxidative stress and mediator for apoptosis.

Dietary cadmium (Cd) can contribute significantly to chronic bioaccumulation and toxicity in aquatic organisms. This contribution needs to be quantified so that the relative importance of waterborne and dietary cadmium exposure pathways can be incorporated into protective water quality guidelines and ecological risk assessments. In this research, the contribution of dietary Cd from a natural periphyton diet to chronic (28 d) bioaccumulation and toxicity in the freshwater amphipod Hyalella azteca was quantified using a mechanistically-based saturation bioaccumulation model. Factors that influence dietary Cd bioavailability such as food type, food form, dietary Cd speciation and concentration were investigated. Assimilation efficiency, ingestion rate and the excretion rate constant of dietary Cd were determined for each of these factors. Food nutrition was also considered. Lastly, model predictions of Cd bioaccumulation and toxicity were compared to measurements of tissue concentration and survival when H. azteca were exposed to metal contaminated water and periphyton collected from lakes in the metal mining region of Rouyn-Noranda, Quebec, Canada. In 28 d laboratory experiments where H. azteca bioaccumulated Cd from water and food, dietary Cd was estimated to contribute markedly (21 – 94 %) to bioaccumulated Cd in H. azteca. Effects on chronic survival were best predicted from body concentration rather than water or food exposure concentration. Assimilation efficiency of dietary Cd differed with food type likely as a result of Cd speciation, but did not differ with Cd concentration or food form. Ingestion rate differed with food form while excretion rate constants were unaffected by dietary Cd bioavailability. Predictions of chronic Cd bioaccumulation in H. azteca exposed to field contaminated samples were robust, however the model did not account for effects of water chemistry on Cd bioaccumulation and is thus constrained in its application. Predictions of chronic survival were over-estimated likely due to the additional toxicity caused by the low nutritional quality of the field contaminated periphyton. This research demonstrated that both waterborne and dietary Cd need to be considered in models that assess chronic risk of exposure and effects to H. azteca.

The U.S. Army Corps of Engineers, Nashville District, maintains ten reservoirs in the Cumberland River Basin in Kentucky and Tennessee, and has been monitoring sediment chemistry in the reservoirs since 1994. The purpose of this study is to evaluate the sediment data collected from the reservoirs from 1994 to 2010 to determine if there are any spatial patterns of the trace elements: arsenic, beryllium, cadmium, chromium, copper, lead, mercury, nickel, and zinc. The results indicated that trace element levels were consistent with national baseline concentrations measured by the U.S. Geological Survey. Center Hill reservoir had the greatest number of trace element concentrations (all except cadmium) that were significantly higher when compared to all other reservoirs. The degree of urbanization in the reservoir basins was based on population density from the 2000 Census and the percentage of developed land using the 2006 national land cover dataset. Aquatic toxicity values were used as a measure of sediment quality. The reservoirs with the worst aquatic toxicity rankings were not the most urban, instead they were the reservoirs with the longest retention times. Therefore, it may be concluded that retention time has a larger effect on Cumberland River Basin sediment concentrations than the type of land use or the degree of urbanization. The results also indicate that it may be prudent to include an evaluation of quality based on aquatic toxicity when monitoring sediment quality, and that when reservoirs are the subject of sediment quality assessments, the consideration of the physical properties of the reservoir, especially the retention time, is essential for a comprehensive evaluation. This may also imply that sediment quality in reservoirs may effectively be regulated by water resource management techniques at the reservoirs that affect retention time.

This thesis deals with the highly potent olfactory mucosa toxicant 2,6-dichlorophenyl methylsulphone (2,6-diClPh-MeSO2) and its non-toxic 2,5-chlorinated isomer (2,5-diClPh-MeSO2). In mice, both substances bind firmly in the olfactory mucosa and the olfactory bulb, which are important components of the sensory system. The 2,6-isomer induces olfactory mucosal necrosis with permanent loss of olfactory neuroepithelium and olfactory nerves. A major objective was to clarify the cause of this isomer-specific toxicity, and to identify which physicochemical characteristics determine the olfactory toxicity. The neurobehavioural toxicity of these substances was also examined. The results revealed a rapid CYP-catalysed covalent binding of 2,6-diClPh-MeSO2 in the rat olfactory mucosa, whereas the 2,5-dichlorinated isomer was not covalently bound. Acute and chronic olfactory mucosal pathology were investigated and compared in rats and mice. Twenty-four hours after dosing to rats, 2,6-diClPh-MeSO2 induced Bowman’s glands necrosis and sloughing of the olfactory epithelium similar to that previously reported in mice. At 3 weeks, however, there were dramatic differences in histological lesions. In mice, large parts of olfactory epithelium were replaced by respiratory-like epithelium. Large, bilateral, fibrous, cartilage and bone containing polyps occluding the lumen were confirmed. In rats, only minor patches of olfactory epithelium were replaced by a metaplastic atypical respiratory-like epithelium. 2,5-diClPh-MeSO2 was non-toxic in rats as well as in mice. In mice, 2,6-diClPh-MeSO2 induced a dose-dependent and long-lasting ( ≥12 weeks) hyperactivity as well as long-lasting maze learning deficits. At 2 weeks hyperactivity and maze learning deficits were observed also in rats. Unexpectedly, 2,5-diClPh-MeSO2 induced hyperactivity that lasted for two weeks. No effect on maze learning was observed with this isomer. No major differences between male and female rats or mice were found. In conclusion, the results show that a CYP-catalysed formation and covalent binding of a reactive 2,6-diClPh-MeSO2-metabolite in the Bowman’s glands precede the high olfactory mucosal toxicity in rodents. As determined by QSAR-modelling, a 2,6-dichlorinated benzene derivative with a large, polar, and strong electron withdrawing substituent in the primary position has the potential of being an olfactory mucosal toxicant. The observed 2,6-diClPh-MeSO2-induced increase in motor activity, and maze learning deficits, were not correlated to the olfactory mucosal lesions. I propose that 2,6-diClPh-MeSO2 causes a direct effect in the brain leading to neurobehaviuoral deficits.

Metal mines release treated effluents that contain a variety of metals, metalloids, and organics into the aquatic environment. A number of metal mine effluents (MMEs) have been found to contribute to adverse effects in fish and benthic invertebrates, such as decreased diversity and density, however the specific causal factors of toxic responses during chronic exposures to the MMEs are often unknown. Therefore, the overall objective of this dissertation was to explore causative and modifying factors of MME toxicity to a resident fish species, the fathead minnow (Pimephales promelas), during chronic, multi-trophic exposures. The representative MME used in this study was the process water effluent (PWE) of a Canadian metal mine, which is released into Junction Creek in Sudbury, Ontario, Canada. Chronic exposure to the MME has been a source of decreased reproductive output in fathead minnows in several previous studies, however, these same studies were not able to determine the potential causal factors of the reproductive impairment. In order to address the overall objective, several laboratory mesocosm studies were conducted, which consisted of three separate components. The first component included exploring several metals (Cu, Ni, and Se; alone and in mixture) that are consistently present in the MME and are known to cause toxicity at fairly low concentrations as potential causes for decreased egg production in fathead minnows. The second component included evaluating the role of decreased food availability (a possible indirect effect of MME in the receiving environment) as a potential cause of decreased egg production in fathead minnows. The third and final component included examining the role of water chemistry [(increased alkalinity and dissolved organic carbon (DOC)] as potential modifying factors of chronic MME toxicity to fathead minnows. In general, my results suggest that the metals present in the MME likely do not contribute directly to decreased reproductive performance in fathead minnows during chronic exposures, under the conditions examined. Instead, the MME appears to decrease food availability, therefore indirectly influence fathead minnow egg production. Furthermore, water chemistry modifications tested in this thesis were not able to entirely mitigate the reproductive effects in fish induced by the MME, although they did improve egg production relative to unmodified MME. Metal concentrations in fish tissues were not influenced by increases to alkalinity or DOC level in the exposure water, suggesting that bioavailability of metals during chronic exposure to metal-mixtures cannot be fully explained based on our understanding of metal complexation with abiotic ligands (inorganic and organic) during single metal or acute exposures. From a regulatory perspective, water chemistry modifications may somewhat improve fathead minnow reproductive performance during chronic exposure to the MME, however the MME would still not be entirely free of effects relative to the uncontaminated water. Future studies should focus on understanding the factors responsible for decreased food availability in MME-impacted aquatic ecosystems, and further explore potential approaches for ameliorating effluent quality.

This work will describe research directed towards the synthesis of anisotropic gold nanoparticles as well as their functionalization and biological applications. The thesis will begin by describing a new technique for the high-yield synthesis of gold nanorods using hydroquinone as a reducing agent. This addresses important limitations of the traditional nanorod synthesis including low yield of gold ions conversion to metallic form and inability to produce rods with longitudinal surface plasmon peak above 850 nm. The use of hydroquinone was also found to improve the synthesis of gold nanowires via the nanorod-seed mediated procedure developed in our lab. The thesis will next present the synthesis of novel starfruit-shaped nanorods, mesorods, and nanowires using a modified nanorod-seed mediated procedure. The starfruit particles displayed increased activity as surface-enhanced Raman spectroscopy (SERS) substrates as compared to smooth structures. Next, a method for the functionalization of gold nanorods using a cationic thiol, 16-mercaptohexadecyltrimethylammonium bromide (MTAB), will be described. By using this thiol, we were able to demonstrate the complete removal of toxic surfactant from the nanorods and were also able to precisely quantify the grafting density of thiol molecules on the nanorod surface through a combination of several analytical techniques. Finally, this thesis will show that MTAB-functionalized nanorods are nontoxic and can be taken up in extremely high numbers into cancer cells. The thesis will conclude by describing the surprising uptake of larger mesorods and nanowires functionalized with MTAB into cells in high quantities.