Radiation impacts on toxicity of cobalt-chromium (CoCr) implant debris.

Particulate and soluble debris are generated by mechanical and non-mechanical degradation of implanted medical devices. Debris containing cobalt and chromium (CoCr) is known to cause adverse biological reactions. Implant-related complications are often diagnosed using radiography, which results in more frequent patient exposure to ionizing radiation. The aim of this study was to evaluate the potential for increased toxicity due to combined radiation and CoCr exposure. This was investigated using a controlled in vitro model consisting of commercially available CoCr debris that was generated from components of hip replacements and human cell lines relevant to the joint environment: endothelial HMEC-1 and synovial SW982. Particle sizes and shapes were heterogenous. Cells tended to internalize smaller particles, as observed by electron microscopy. Indicators of toxicity were measured after short (24 h after radiation) or extended (12-14 d after radiation) exposure timelines. In the short-term, CoCr reduced cell viability, increased apoptosis, and increased oxidative stress. The effects of radiation were not apparent until the timeline was extended. CoCr and radiation reduced cell survival, with both additive and synergistic effects. Mechanisms for reduced survival included rapid cell death caused by CoCr and senescence caused by radiation. In conclusion, results showed combined toxicological effects of CoCr and radiation at the doses and timelines used for this in vitro model. These observations warrant further investigation using other experimental models to determine translational impact.

Unraveling Metabolic and Proteomic Features in Soybean Plants in Response to Copper Hydroxide Nanowires Compared to a Commercial Fertilizer.

Mechanistic understanding of the interaction of copper-based nanomaterials with crops is crucial for exploring their application in precision agriculture and their implications on plant health. We investigated the biological response of soybean (Glycine max) plants to the foliar application of copper hydroxide nanowires (CNWs) at realistic exposure concentrations. A commercial copper based-fungicide (Kocide), dissolved copper ions, and untreated controls were used for comparison to identify unique features at physiological, cellular, and molecular levels. After 32 d of exposure to CNW (0.36, 1.8, and 9 mg CNW/plant), the newly developed tissues accumulated significantly high levels of Cu (18-60 µg/g) compared to Kocide (10 µg/g); however, the rate of Cu translocation from the site of CNW treatment to other tissues was slower compared to other Cu treatments. Like Kocide, CNW exposure at medium and high doses altered Co, Mn, Zn, and Fe accumulation in the tissues and enhanced photosynthetic activities. The proteomic and metabolomic analyses of leaves from CNW-treated soybean plants suggest a dose-dependent response, resulting in the activation of major biological processes, including photosynthesis, energy production, fatty acid metabolism, lignin biosynthesis, and carbohydrate metabolism. In contrast to CNW treatments, Kocide exposure resulted in increased oxidative stress response and amino acid metabolism activation.

Metabolomic Responses of Green Alga Chlamydomonas reinhardtii Exposed to Sublethal Concentrations of Inorganic and Methylmercury.

Metabolomics characterizes low-molecular-weight molecules involved in different biochemical reactions and provides an integrated assessment of the physiological state of an organism. By using liquid chromatography-mass spectrometry targeted metabolomics, we examined the response of green alga Chlamydomonas reinhardtii to sublethal concentrations of inorganic mercury (IHg) and monomethylmercury (MeHg). We quantified the changes in the levels of 93 metabolites preselected based on the disturbed metabolic pathways obtained in a previous transcriptomics study. Metabolites are downstream products of the gene transcription; hence, metabolite quantification provided information about the biochemical status of the algal cells exposed to Hg compounds. The results showed that the alga adjusts its metabolism during 2 h exposure to 5 × 10-9 and 5 × 10-8 mol L-1 IHg and MeHg by increasing the level of various metabolites involved in amino acid and nucleotide metabolism, photorespiration, and tricarboxylic acid (TCA) cycle, as well as the metabolism of fatty acids, carbohydrates, and antioxidants. Most of the metabolic perturbations in the alga were common for IHg and MeHg treatments. However, the exposure to IHg resulted in more pronounced perturbations in the fatty acid and TCA metabolism as compared with the exposure to MeHg. The observed metabolic perturbations were generally consistent with our previously published transcriptomics results for C. reinhardtii exposed to the comparable level of IHg and MeHg. The results highlight the potential of metabolomics for toxicity evaluation, especially to detect effects at an early stage of exposure prior to their physiological appearance.

Metabolomics for early detection of stress in freshwater alga Poterioochromonas malhamensis exposed to silver nanoparticles.

Silver nanoparticles (AgNPs) are one of the most used engineered nanomaterials. Despite progress in assessing their environmental implications, knowledge gaps exist concerning the metabolic perturbations induced by AgNPs on phytoplankton, essential organisms in global biogeochemical cycles and food-web dynamics. We combine targeted metabolomics, biouptake and physiological response studies to elucidate metabolic perturbations in alga Poterioochromonas malhamensis induced by AgNPs and dissolved Ag. We show time-dependent perturbation of the metabolism of amino acids, nucleotides, fatty acids, tricarboxylic acids, photosynthesis and photorespiration by both Ag-treatments. The results suggest that dissolved Ag ions released by AgNPs are the major toxicity driver; however, AgNPs internalized in food vacuoles contributed to the perturbation of amino acid metabolism, TCA cycle and oxidative stress. The metabolic perturbations corroborate the observed physiological responses. We highlight the potential of metabolomics as a tool for understanding the molecular basis for these metabolic and physiological changes, and for early detection of stress.

Effect of Metalloid and Metal Oxide Nanoparticles on Fusarium Wilt of Watermelon.

This study explored the use of foliar sprays with nanoparticles (NP) of B, CuO, MnO, SiO, TiO, and ZnO to protect watermelon against Fusarium wilt. Leaves of young watermelon plants were sprayed (1 to 2 ml per plant) with NP suspensions (500 to 1,000 µg/ml) and were planted in potting mix infested with Fusarium oxysporum f. sp. niveum. In five of eight greenhouse experiments, CuO NP suppressed disease and, in six of eight experiments, CuO NP increased biomass or yield more than in untreated controls or other tested NP. More root Cu was detected in CuO NP-treated plants than other treatments (P = 0.015). In Griswold, CT, plants treated with CuO NP yielded 39% more fruit than untreated controls. In Hamden, CT, treatment with CuO NP produced 53% more fruit when compared with controls (P = 0.02) and was superior to other Cu fungicides. Gene expression in watermelon roots revealed strong upregulation of polyphenol oxidase (PPO) and PR1 genes when CuO NP and F. oxysporum f. sp. niveum were both present. Enzymatic assays for PPO supported the gene expression results. CuO NP may serve as a highly effective delivery agent for this micronutrient to suppress disease.

Nanoparticle interactions with co-existing contaminants: joint toxicity, bioaccumulation and risk.

With their growing production and application, engineered nanoparticles (NPs) are increasingly discharged into the environment. The released NPs can potentially interact with pre-existing contaminants, leading to biological effects (bioaccumulation and/or toxicity) that are poorly understood. Most studies on NPs focus on single analyte exposure; the existing literature on joint toxicity of NPs and co-existing contaminants is rather limited but beginning to develop rapidly. This is the first review paper evaluating the current state of knowledge regarding the joint effects of NPs and co-contaminants. Here, we review: (1) methods for investigating and evaluating joint effects of NPs and co-contaminants; (2) simultaneous toxicities from NPs co-exposed with organic contaminants, metal/metalloid ions, dissolved organic matter (DOM), inorganic ligands and additional NPs; and (3) the influence of NPs co-exposure on the bioaccumulation of organic contaminants and heavy metal ions, as well as the influence of contaminants on NPs bioaccumulation. In addition, future research needs are discussed so as to better understand risk associated with NPs-contaminant co-exposure.

A Collaborative Study: Determination of Mycotoxins in Corn, Peanut Butter, and Wheat Flour Using Stable Isotope Dilution Assay (SIDA) and Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS).

A collaborative study was conducted to evaluate stable isotope dilution assay (SIDA) and LC-MS/MS for the simultaneous determination of aflatoxins B1, B2, G1, and G2; deoxynivalenol; fumonisins B1, B2, and B3; ochratoxin A; HT-2 toxin; T-2 toxin; and zearalenone in foods. Samples were fortified with 12 13C uniformly labeled mycotoxins (13C-IS) corresponding to the native mycotoxins and extracted with acetonitrile/water (50:50 v/v), followed by centrifugation, filtration, and LC-MS/MS analysis. In addition to certified reference materials, the six participating laboratories analyzed corn, peanut butter, and wheat flour fortified with the 12 mycotoxins at concentrations ranging from 1.0 to 1000 ng/g. Using their available LC-MS/MS platform, each laboratory developed in-house instrumental conditions for analysis. The majority of recoveries ranged from 80 to 120% with relative standard derivations (RSDs) <20%. Greater than 90% of the average recoveries of the participating laboratories were in the range of 90-110%, with repeatability RSDr (within laboratory) < 10% and reproducibility RSDR (among laboratory) < 15%. All Z scores of the results of certified reference materials were between -2 and 2. Using 13C-IS eliminated the need for matrix-matched calibration standards for quantitation, simplified sample preparation, and achieved simultaneous identification and quantitation of multiple mycotoxins in a simple LC-MS/MS procedure.

Weathering in soil increases nanoparticle CuO bioaccumulation within a terrestrial food chain.

This study evaluates the bioaccumulation of unweathered (U) and weathered (W) CuO in NP, bulk and ionic form (0-400 mg/kg) by lettuce exposed for 70 d in soil co-contaminated with field incurred chlordane. To evaluate CuO trophic transfer, leaves were fed to crickets (Acheta domestica) for 15 d, followed by insect feeding to lizards (Anolis carolinensis). Upon weathering, the root Cu content of the NP treatment increased 214% (327 ± 59.1 mg/kg) over unaged treatment. Cu root content decreased in bulk and ionic treatments from 70-130 mg/kg to 13-26 mg/kg upon aging in soil. Micro X-ray fluorescence (µ-XRF) analysis of W-NP-exposed roots showed a homogenous distribution of Cu (and Ca) in the tissues. Additionally, micro X-ray absorption near-edge (µ-XANES) analysis of W-NP-exposed roots showed near complete transformation of CuO to Cu (I)-sulfur and oxide complexes in the tissues, whereas in unweathered treatment, most root Cu remained as CuO. The expression level of nine genes involved in Cu transport shows that the mechanisms of CuO NPs (and bulk) response/accumulation are different than ionic Cu. The chlordane accumulation by lettuce upon co-exposure to CuO NPs significantly increased upon weathering. Conversely, bulk and ionic exposures decreased pesticide accumulation by plant upon weathering. The Cu cricket fecal content from U-NP-exposed insects was significantly greater than the bulk or ion treatments, suggesting a higher initial NP accumulation followed by significantly greater elimination during depuration. In the lizard, Cu content in the intestine, body and head did not differ as a function of weathering. This study demonstrates that CuO NPs may undergo transformation processes in soil upon weathering that subsequently impact NPs availability in terrestrial food chains.

Molecular Response of Crop Plants to Engineered Nanomaterials.

Functional toxicology has enabled the identification of genes involved in conferring tolerance and sensitivity to engineered nanomaterial (ENM) exposure in the model plant Arabidopsis thaliana (L.) Heynh. Several genes were found to be involved in metabolic functions, stress response, transport, protein synthesis, and DNA repair. Consequently, analysis of physiological parameters, metal content (through ICP-MS quantification), and gene expression (by RT-qPCR) of A. thaliana orthologue genes were performed across different plant species of agronomic interest to highlight putative biomarkers of exposure and effect related to ENMs. This approach led to the identification of molecular markers in Solanum lycopersicum L. and Cucurbita pepo L. (tomato and zucchini) that might not only indicate exposure to ENMs (CuO, CeO2, and La2O3) but also provide mechanistic insight into response to these materials. Through Gene Ontology (GO) analysis, the target genes were mapped in complex interatomic networks representing molecular pathways, cellular components, and biological processes involved in ENM response. The transcriptional response of 38 (out of 204) candidate genes studied varied according to particle type, size, and plant species. Importantly, some of the genes studied showed potential as biomarkers of ENM exposure and effect and may be useful for risk assessment in foods and in the environment.

Soil organic matter influences cerium translocation and physiological processes in kidney bean plants exposed to cerium oxide nanoparticles.

Soil organic matter plays a major role in determining the fate of the engineered nanomaterials (ENMs) in the soil matrix and effects on the residing plants. In this study, kidney bean plants were grown in soils varying in organic matter content and amended with 0-500mg/kg cerium oxide nanoparticles (nano-CeO2) under greenhouse condition. After 52days of exposure, cerium accumulation in tissues, plant growth and physiological parameters including photosynthetic pigments (chlorophylls and carotenoids), net photosynthesis rate, transpiration rate, and stomatal conductance were recorded. Additionally, catalase and ascorbate peroxidase activities were measured to evaluate oxidative stress in the tissues. The translocation factor of cerium in the nano-CeO2 exposed plants grown in organic matter enriched soil (OMES) was twice as the plants grown in low organic matter soil (LOMS). Although the leaf cover area increased by 65-111% with increasing nano-CeO2 concentration in LOMS, the effect on the physiological processes were inconsequential. In OMES leaves, exposure to 62.5-250mg/kg nano-CeO2 led to an enhancement in the transpiration rate and stomatal conductance, but to a simultaneous decrease in carotenoid contents by 25-28%. Chlorophyll a in the OMES leaves also decreased by 27 and 18% on exposure to 125 and 250mg/kg nano-CeO2. In addition, catalase activity increased in LOMS stems, and ascorbate peroxidase increased in OMES leaves of nano-CeO2 exposed plants, with respect to control. Thus, this study provides clear evidence that the properties of the complex soil matrix play decisive roles in determining the fate, bioavailability, and biological transport of ENMs in the environment.

Carbon Nanomaterials in Agriculture: A Critical Review.

There has been great interest in the use of carbon nano-materials (CNMs) in agriculture. However, the existing literature reveals mixed effects from CNM exposure on plants, ranging from enhanced crop yield to acute cytotoxicity and genetic alteration. These seemingly inconsistent research-outcomes, taken with the current technological limitations for in situ CNM detection, present significant hurdles to the wide scale use of CNMs in agriculture. The objective of this review is to evaluate the current literature, including studies with both positive and negative effects of different CNMs (e.g., carbon nano-tubes, fullerenes, carbon nanoparticles, and carbon nano-horns, among others) on terrestrial plants and associated soil-dwelling microbes. The effects of CNMs on the uptake of various co-contaminants will also be discussed. Last, we highlight critical knowledge gaps, including the need for more soil-based investigations under environmentally relevant conditions. In addition, efforts need to be focused on better understanding of the underlying mechanism of CNM-plant interactions.

Cerium Biomagnification in a Terrestrial Food Chain: Influence of Particle Size and Growth Stage.

Mass-flow modeling of engineered nanomaterials (ENMs) indicates that a major fraction of released particles partition into soils and sediments. This has aggravated the risk of contaminating agricultural fields, potentially threatening associated food webs. To assess possible ENM trophic transfer, cerium accumulation from cerium oxide nanoparticles (nano-CeO2) and their bulk equivalent (bulk-CeO2) was investigated in producers and consumers from a terrestrial food chain. Kidney bean plants (Phaseolus vulgaris var. red hawk) grown in soil contaminated with 1000-2000 mg/kg nano-CeO2 or 1000 mg/kg bulk-CeO2 were presented to Mexican bean beetles (Epilachna varivestis), which were then consumed by spined soldier bugs (Podisus maculiventris). Cerium accumulation in plant and insects was independent of particle size. After 36 days of exposure to 1000 mg/kg nano- and bulk-CeO2, roots accumulated 26 and 19 µg/g Ce, respectively, and translocated 1.02 and 1.3 µg/g Ce, respectively, to shoots. The beetle larvae feeding on nano-CeO2 exposed leaves accumulated low levels of Ce since ∼98% of Ce was excreted in contrast to bulk-CeO2. However, in nano-CeO2 exposed adults, Ce in tissues was higher than Ce excreted. Additionally, Ce content in tissues was biomagnified by a factor of 5.3 from the plants to adult beetles and further to bugs.

Environmental Effects of Nanoceria on Seed Production of Common Bean (Phaseolus vulgaris): A Proteomic Analysis.

The rapidly growing literature on the response of edible plants to nanoceria has provided evidence of its uptake and bioaccumulation, which delineates a possible route of entry into the food chain. However, little is known about how the residing organic matter in soil may affect the bioavailability and resulting impacts of nanoceria on plants. Here, we examined the effect of nanoceria exposure (62.5-500 mg/kg) on kidney bean (Phaseolus vulgaris) productivity and seed quality as a function of soil organic matter content. Cerium accumulation in the seeds produced from plants in organic matter enriched soil showed a dose-dependent increase, unlike in low organic matter soil treatments. Seeds obtained upon nanoceria exposure in soils with higher organic matter were more susceptible to changes in nutrient quality. A quantitative proteomic analysis of the seeds produced upon nanoceria exposure provided evidence for upregulation of stress-related proteins at 62.5 and 125 mg/kg nanoceria treatments. Although the plants did not exhibit overt toxicity, the major seed proteins primarily associated with nutrient storage (phaseolin) and carbohydrate metabolism (lectins) were significantly down-regulated in a dose dependent manner upon nanoceria exposure. This study thus suggests that nanoceria exposures may negatively affect the nutritional quality of kidney beans at the cellular and molecular level. More confirmatory studies with nanoceria along different species using alternative and orthogonal "omic" tools are currently under active investigation, which will enable the identification of biomarkers of exposure and susceptibility.

Monitoring the environmental effects of CeO2 and ZnO nanoparticles through the life cycle of corn (Zea mays) plants and in situ µ-XRF mapping of nutrients in kernels.

Information about changes in physiological and agronomic parameters through the life cycle of plants exposed to engineered nanoparticles (NPs) is scarce. In this study, corn (Zea mays) plants were cultivated to full maturity in soil amended with either nCeO2 or nZnO at 0, 400, and 800 mg/kg. Gas exchange was monitored every 10 days, and at harvest, bioaccumulation of Ce and Zn in tissues was determined by ICP-OES/MS. The effects of NPs exposure on nutrient concentration and distribution in ears were also evaluated by ICP-OES and µ-XRF. Results showed that nCeO2 at both concentrations did not impact gas exchange in leaves at any growth stage, while nZnO at 800 mg/kg reduced net photosynthesis by 12%, stomatal conductance by 15%, and relative chlorophyll content by 10% at day 20. Yield was reduced by 38% with nCeO2 and by 49% with nZnO. Importantly, µ-XRF mapping showed that nCeO2 changed the allocation of calcium in kernels, compared to controls. In nCeO2 treated plants, Cu, K, Mn, and Zn were mainly localized at the insertion of kernels into cobs, but Ca and Fe were distributed in other parts of the kernels. Results showed that nCeO2 and nZnO reduced corn yield and altered quality of corn.

Particle-size dependent accumulation and trophic transfer of cerium oxide through a terrestrial food chain.

The accumulation and trophic transfer of nanoparticle (NP) or bulk CeO2 through a terrestrial food chain was evaluated. Zucchini (Cucurbita pepo L.) was planted in soil with 0 or 1228 µg/g bulk or NP CeO2. After 28 d, zucchini tissue Ce content was determined by ICP-MS. Leaf tissue from each treatment was used to feed crickets (Acheta domesticus). After 14 d, crickets were analyzed for Ce content or were fed to wolf spiders (family Lycosidae). NP CeO2 significantly suppressed flower mass relative to control and bulk treatments. The Ce content of zucchini was significantly greater when exposure was in the NP form. The flowers, leaves, stems, and roots of zucchini exposed to bulk CeO2 contained 93.3, 707, 331, and 119,000 ng/g, respectively; NP-exposed plants contained 153, 1510, 479, and 567 000 ng/g, respectively. Crickets fed NP CeO2-exposed zucchini leaves contained significantly more Ce (33.6 ng/g) than did control or bulk-exposed insects (15.0-15.2 ng/g). Feces from control, bulk, and NP-exposed crickets contained Ce at 248, 393, and 1010 ng/g, respectively. Spiders that consumed crickets from control or bulk treatments contained nonquantifiable Ce; NP-exposed spiders contained Ce at 5.49 ng/g. These findings show that NP CeO2 accumulates in zucchini at greater levels than equivalent bulk materials and that this greater NP intake results in trophic transfer and possible food chain contamination.

Exposure of cerium oxide nanoparticles to kidney bean shows disturbance in the plant defense mechanisms.

Overwhelming use of engineered nanoparticles demands rapid assessment of their environmental impacts. The transport of cerium oxide nanoparticles (nCeO2) in plants and their impact on cellular homeostasis as a function of exposure duration is not well understood. In this study, kidney bean plants were exposed to suspensions of ∼ 8 ± 1 nm nCeO2 (62.5 to 500 mg/L) for 15 days in hydroponic conditions. Plant parts were analyzed for cerium accumulation after one, seven, and 15 days of nCeO2 exposure. The primary indicators of stress like lipid peroxidation, antioxidant enzyme activities, total soluble protein and chlorophyll contents were studied. Cerium in tissues was localized using scanning electron microscopy and synchrotron µ-XRF mapping, and the chemical forms were identified using µ-XANES. In the root epidermis, cerium was primarily shown to exist as nCeO2, although a small fraction (12%) was biotransformed to Ce(III) compound. Cerium was found to reach the root vascular tissues and translocate to aerial parts with time. Upon prolonged exposure to 500 mg nCeO2/L, the root antioxidant enzyme activities were significantly reduced, simultaneously increasing the root soluble protein by 204%. In addition, leaf's guaiacol peroxidase activity was enhanced with nCeO2 exposure in order to maintain cellular homeostasis.

Applications of synchrotron µ-XRF to study the distribution of biologically important elements in different environmental matrices: a review.

Environmental matrices including soils, sediments, and living organisms are reservoirs of several essential as well as non-essential elements. Accurate qualitative and quantitative information on the distribution and interaction of biologically significant elements is vital to understand the role of these elements in environmental and biological samples. Synchrotron micro-X-ray fluorescence (µ-SXRF) allows in situ mapping of biologically important elements at nanometer to sub-micrometer scale with high sensitivity, negligible sample damage and enable tuning of the incident energy as desired. Beamlines in the synchrotron facilities are rapidly increasing their analytical versatility in terms of focusing optics, detector technologies, incident energy, and sample environment. Although extremely competitive, it is now feasible to find stations offering complimentary techniques like micro-X-ray diffraction (µ-XRD) and micro-X-ray absorption spectroscopy (µ-XAS) that will allow a more complete characterization of complex matrices. This review includes the most recent literature on the emerging applications and challenges of µ-SXRF in studying the distribution of biologically important elements and manufactured nanoparticles in soils, sediments, plants, and microbes. The advantages of using µ-SXRF and complimentary techniques in contrast to conventional techniques used for the respective studies are discussed.

Interaction of nanoparticles with edible plants and their possible implications in the food chain.

The uptake, bioaccumulation, biotransformation, and risks of nanomaterials (NMs) for food crops are still not well understood. Very few NMs and plant species have been studied, mainly at the very early growth stages of the plants. Most of the studies, except one with multiwalled carbon nanotubes performed on the model plant Arabidopsis thaliana and another with ZnO nanoparticles (NPs) on ryegrass, reported the effect of NMs on seed germination or 15-day-old seedlings. Very few references describe the biotransformation of NMs in food crops, and the possible transmission of the NMs to the next generation of plants exposed to NMs is unknown. The possible biomagnification of NPs in the food chain is also unknown.