Kinetics of phytoremediation of petroleum hydrocarbon contaminated soil.

Earthmaster and the University of Waterloo have successfully developed plant growth promoting rhizobacteria (PGPR) - Enhanced Phytoremediation Systems (PEPSystems™) which have been deployed across Canada for the treatment of soil contaminated with petroleum hydrocarbons (PHCs), including CCME fractions F2 and F3. A challenge with phytoremediation is to predict the length of time to remediate a site so that site owners will be inclined to use the technology. In previous field trials of PEPSystems, it was determined that PHC was mostly degraded by microbes in the rhizosphere, following first-order exponential decay kinetics. Using new PEPSystems data collected from multiple commercial remediation sites across Western Canada, the kinetic equations of PHC decay were tested to determine if remediation time was accurately predicted. In general, when compared to the predicted time to remediation endpoint, data from recent commercial field applications showed that 35% and 20% less time was needed to reach remediation endpoints for fractions F2 and F3, respectively. As a result, the predictive kinetic equation for fraction F2 degradation was updated to reflect current remediation outcomes. Insufficient data were available to update the F3 equation. Being able to more accurately predict remediation timelines will enhance the value and utilization of PEPSystems.

Motorized dirt bike injuries in children.

BACKGROUND: The number of dirt bike injuries in children in the United States is increasing and poses a public health problem. OBJECTIVE: The purpose of our study was to identify the imaging patterns of dirt bike injuries in children and associations with morbidity and mortality. METHODS: The study included 85 children (83 boys, 2 girls) <18 year of age (mean age 12.3 years, standard deviation 3 years) with dirt bike injury treated at a tertiary care pediatric hospital. Imaging studies and hospital medical records were reviewed. Outcomes were classified into the following categories: short-term disability, long-term disability or no follow-up available. Imaging studies were reviewed for head, torso, and extremity injuries. One-tailed z test for two proportions was used to determine significant differences between various proportions. Chi-square test with Yates correction was used to determine the significance of long-term disability with injury type. RESULTS: Long bone fractures were the most common injuries. Lower extremity fractures accounted for 79% of extremity fractures and were significantly more common than upper extremity fractures (p = 0.001). Head injuries included fractures (n = 9), brain contusion (n = 5), and meningeal hemorrhage (n = 2). Head injury was associated with long-term disability (p < 0.0001). All torso injuries were solitary. CONCLUSIONS: Long-term disability was associated with head injuries but not with torso or extremity injuries. Lower extremity injuries were significantly more common than upper extremity injuries. Torso solid organ injuries were uniformly solitary.

Atypical antipsychotics for the treatment of ICU delirium.

Delirium is commonly described in critically ill patients as 1 factor contributing to increased length of intensive care unit and hospital stay, secondary complications, and increased mortality. Initial screening tools for delirium in hospitalized patients are generally easy to use; however, many centers have struggled with implementing these tools in a consistent and systematic manner. Haloperidol has traditionally been prescribed as the primary agent of choice for the treatment of delirium in critically ill patients. Clinicians have been challenged to consider alternative agents due to adverse effects such as extrapyramidal symptoms, QTc prolongation, and possible torsades de pointes with haloperidol use. The atypical antipsychotics are attractive alternatives to haloperidol with improved safety profiles but are flawed by limited data to support dosing and efficacy in this patient population. Future studies that provide large, prospective, double-blinded, placebo-controlled data to support the implementation of these agents as standard therapy over haloperidol are needed.

Bioavailability of copper and zinc in mining soils.

The soil-contact exposure pathway can be the main driver of ecological risk assessments. There is currently no standard method to measure bioavailability of metals in soil to ecological receptors, yet the influence of metal bioavailability on toxicity has been known for decades and is a major factor influencing risk to ecological receptors. Bioavailability is to a large degree governed by varying soil characteristics within and among sites, yet ecological screening benchmarks are often derived on a total-concentration basis. We compared a calcium chloride (CaCl2) extraction, cyclodextrin extraction, simulated earthworm gut (SEG) test, earthworm kinetic bioaccumulation test, and metal residues in plant tissues with a battery of invertebrate and toxicity tests using mining soils consisting of high organic-matter content cocontaminated with copper (Cu) and zinc (Zn). Earthworm (Eisenia andrei) tissue concentrations of Cu and Zn were regulated and were not predictive of invertebrate toxicity. All chemical measures of bioavailability correlated with several biological responses; however, CaCl2-extractable Cu and SEG-extractable Cu and Zn best predicted effects to E. andrei. Total Cu concentrations in soil best correlated with effects to plants. Overall, a chemical measure was the best predictor of toxicity to each organism compared with biological measures, although the exact measure was dependent on organism and end point. Chemical-extraction techniques provide relatively quick, inexpensive indicators of essential metal bioavailability compared with biological measures; however, no single measure was indicative of all effects to all organisms.

Photosynthetic redox imbalance influences flavonoid biosynthesis in Lemna gibba.

Plants accumulate flavonoids in response to a myriad of environmental challenges, especially when exposed to ultraviolet (UV) radiation or situations causing oxidative stress. However, the origin and nature of the signal triggering their accumulation remain obscure. In this study, a group of flavonoids belonging to the flavone class was identified in Lemna gibba (duckweed). These flavones accumulated upon exposure to UV radiation, low temperature, copper and the photosynthetic electron transport (PET) inhibitors 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) and 1,2-dihydroxyanthraquinone (DHATQ). All of these stressors were also shown to promote PET chain (PETC) reduction; however, in the co-presence of 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU) or a light regime that oxidized the PETC, flavonoid accumulation ceased. Chloroplast-derived reactive oxygen species (ROS) were not associated with all of the stress conditions that promoted both PETC reduction and flavonoid synthesis, indicating that ROS were not a strict requisite for flavonoid accumulation. Transcripts for the flavonoid biosynthetic genes, chalcone synthase (CHS) and chalcone isomerase, were similarly responsive to the PETC redox state, as were a panel of transcripts revealed by differential display PCR. Collectively, these results provide evidence that PETC redox status is one of the factors affecting flavonoid biosynthesis.

Imaging findings in 512 children following all-terrain vehicle injuries.

BACKGROUND: Injuries related to all-terrain vehicle (ATV) use by children have increased in recent years, and the pattern of these injuries is not well known among radiologists. OBJECTIVE: Our purpose was to identify different radiologically diagnosed injuries in children suffering ATV-related trauma and determine associations among various injuries as well as between injuries and outcome. MATERIALS AND METHODS: The study included 512 consecutive children suffering from ATV injuries treated at a tertiary care pediatric hospital. All imaging studies were reviewed and correlated with injury frequency and outcome using multivariate analysis. RESULTS: Head injuries occurred in 244 children (48%) and in five of six deaths. Calvarial skull fractures occurred in 104 children and were associated with brain, subdural and epidural injuries. Brain and orbit injuries were associated with long-term disability. A total of 227 extremity fractures were present in 172 children (34%). The femur was the most commonly fractured bone. Nine children had partial foot amputations. Multiorgan injuries occurred in nearly half of the 97 children with torso injuries. Determinants for long-term disability or death were head injuries (odds ratio 3.4) and extremity fractures (odds ratio 3.3). CONCLUSION: Head and extremity injuries are the two most common injuries in children suffering ATV injuries and are associated with long-term disability. ATV use by children is dangerous and is a significant threat to child safety.

Examination of the mechanism of phenanthrenequinone toxicity to Vibrio fischeri: evidence for a reactive oxygen species-mediated toxicity mechanism.

Phenanthrenequinone (PHQ) is a photoproduct of phenanthrene, one of the most prevalent polycyclic aromatic hydrocarbons in the environment. Phenanthrenequinone is a compound of substantial interest, because its toxicity can be much greater than its parent chemical to aquatic organisms. The toxicity mechanisms of PHQ to the luminescent marine bacterium Vibrio fischeri were examined in the present study. Phenanthrenequinone can redox cycle in bacterial cells and transfer electrons to O2, enhancing the production of superoxide (O*2-), hydrogen peroxide (H2O2), and other reactive oxygen species (ROS). Exposure of cells to PHQ increased activity of superoxide dismutase (SOD), which detoxifies the ROS superoxide. Concentrations of PHQ that induced the production of H2O2 and other ROS, as well as the elevated levels of Fe-SOD, were correlated with its toxicity as measured by luminescence. Furthermore, toxicity of PHQ to V. fischeri was lowered under the anaerobic conditions, suggesting that the absence of oxygen, which would limit the production of ROS, alleviated toxicity of PHQ. Thus, a ROS-mediated toxicity mechanism of PHQ is highly implicated by in the present study.

Assessment of mixture toxicity of copper, cadmium, and phenanthrenequinone to the marine bacterium Vibrio fischeri.

Transition metals and polycyclic aromatic hydrocarbons (PAHs) are cocontaminants at many sites. Contaminants in mixtures are known to interact with biological systems in ways that can greatly alter the toxicity of individual compounds. The toxicities (individually and as mixtures) of copper (Cu), a redox-active metal; cadmium (Cd), a nonredox active metal; and phenanthrenequinone (PHQ), a redox-active oxygenated PAH, were examined using the bioluminescent bacterium Vibrio fischeri. We found that the cotoxicity of Cu/PHQ was dependent on the ratio of concentrations of each chemical in the mixture. Different interaction types (synergism, antagonism, and additivity) were observed with different combinations of these toxicants. The interaction types changed from antagonism at a low Cu to PHQ ratio (1:4), to additive at an intermediate Cu to PHQ ratio (2:3), to synergistic at higher Cu to PHQ ratios (3:2 and 4:1). In contrast to Cu/PHQ mixtures, the cotoxicity of Cd/PHQ did not change at different mixture ratios and was found for the most part to be additive. For the individual chemicals and their mixtures, reactive oxygen species (ROS) production was observed in V. fischeri, suggesting that individual and mixture toxicity of Cu, Cd, and PHQ to V. fischeri involves ROS-related mechanisms. This study shows that mixture ratios can alter individual chemical toxicity, and should be taken into account in risk assessment.

The effects of far-red light on plant growth and flavonoid accumulation in Brassica napus in the presence of ultraviolet B radiation.

Flavonoid induction is regulated by complex signal transduction pathways involving cryptochrome, phytochrome and UVB photoreceptors. Previously, we identified the UVB-inducible flavonoids in Brassica napus cv. Topas leaves and showed that UVA affected accumulation of the quercetin (Q) and kaempferol (K) glycosides (Wilson et al. [2000] Photochem. Photobiol. 73, 678-684). In this study, we examined the effects of far-red light (FR, 700-780 nm) on UVB-mediated flavonoid accumulation in B. napus. Plants were grown under photosynthetically active radiation (PAR, 400-700 nm, 150 micromol m(-2) s(-1)) plus a moderate level of FR (35 micromol m(-2) s(-1)) for 14 days, and then transferred to five different irradiation regimes (PAR +/- [UVA + UVB] + moderate, intermediate or low fluence FR) for 4 days. Kinetics of flavonoid accumulation were assessed via HPLC. Accumulation of flavonoids, in general, was suppressed by increasing the amount of FR in the spectrum. Furthermore, addition of UVB (290-320 nm) to the spectrum altered the flavonoid composition by causing significant changes in the quantities of individual flavonoids. The relative levels of acylated K glycosides were diminished whereas the relative levels of nonacylated Q glycosides increased dramatically. With UVB exposure there was a five-fold increase in the Q:K ratio. In contrast, increasing the level of FR in the presence of UVB decreased the Q:K ratio by half.

Effect of copper on the cytotoxicity of phenanthrene and 9,10-phenanthrenequinone to the human placental cell line, JEG-3.

The trophoblast cell line, JEG-3, was used to study the cytotoxicity of phenanthrene, 9,10-phenanthrenequinone (PHEQ), anthracene, and 9,10-anthracenedione alone and with copper. The endpoints were the capacity of cultures to reduce alamar Blue (AB), a measure of energy metabolism, and to convert carboxyfluorescein diacetate acetoxymethyl ester (CFDA AM) to carboxyfluorescein, an indication of membrane integrity. Only PHEQ elicited a cytotoxic response. PHEQ caused a concentration-dependent decline in AB but not in CFDA AM readings, suggesting an impairment to energy metabolism. In the presence of copper, PHEQ concentration-response curves were shifted to the left for AB and were obtained with CFDA AM. The Cu/PHEQ synergy is attributed to an increase in redox cycling and production of reactive oxygen species (ROS), which overwhelm antioxidant defenses, damaging energy metabolism first and then membrane integrity. The impermeable copper chelator, bathocuproine, reduced the PHEQ/copper interaction, but the permeable chelator, neocuproine, and copper together were cytotoxic.

Assessment of the toxicity of mixtures of nickel or cadmium with 9,10-phenanthrenequinone to Daphnia magna: impact of a reactive oxygen-mediated mechanism with different redox-active metals.

Recently, we showed that reactive oxygen species (ROS) formation was involved in the toxicity of the redox-active metal Cu and mixtures of Cu plus a photomodified polycyclic aromatic hydrocarbon (PAH), phenanthrenequinone (PHQ), to Daphnia magna. It is unknown, however, if similar results can be observed for metals with lower or no redox activity and their mixtures with PHQ. In the present study using D. magna, the toxicity of Ni, a weakly redox-active metal, and of Cd, a non-redox active metal, was examined with or without PHQ. The abilities of Ni, Cd, PHQ, and binary mixtures of metal plus PHQ to generate ROS were measured using a 2',7'-dichlorofluorescein fluorescence assay. The results were compared with the results of Cu and mixtures of Cu plus PHQ from a recent study by our group. The order of metal toxicity to D. magna was found to be Cd > or = Cu > Ni. As with Cu/PHQ mixtures, synergistic toxicity was observed for mixtures of Ni and PHQ, whereas additive toxicity was observed for mixtures of Cd and PHQ. Alone, PHQ had no impact on ROS levels in D. magna. Nickel alone caused elevated ROS, which was further enhanced in the presence of PHQ. Neither Cd nor Cd/PHQ mixtures increased ROS production. Attenuation of toxicity and ROS production was observed in response to treatment with low concentrations of L-ascorbic acid. These results indicate potential toxic interactions between metals and modified PAHs. With redox-active metals, such as Cu and Ni, and modified PAHs, such as PHQ, these interactions can involve ROS formation.

A predictive quantitative structure-activity relationship model for the photoinduced toxicity of polycyclic aromatic hydrocarbons to Daphnia magna with the use of factors for photosensitization and photomodification.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants that readily absorb environmentally relevant solar ultraviolet radiation. On absorption of a photon, photoinduced toxicity of PAHs is manifested through photosensitization and photomodification. Both of these processes occur under environmentally relevant levels of actinic radiation. An empirical quantitative structure-activity relationship model previously developed was explanatory of photoinduced toxicity of 16 PAHs in Lemna gibba (duckweed). This model was found to be predictive of toxicity to Vibrio fischeri. The L. gibba quantitative structure-activity relationship showed that a photosensitization factor and a photomodification factor could be combined to describe photoinduced toxicity. To further examine this model, we assessed whether it could be applied to Daphnia magna (water flea), a key bioindicator species in aquatic ecosystems. Toxicity was assessed as median effective concentration and median effective time for immobility. As with L. gibba and V. fischeri, neither the photosensitization factor nor the photomodification factor alone correlated to toxicity in D. magna. However, a photosensitization factor modified for D. magna exhibited a correlation to toxicity (r2 = 0.86), which was modestly improved when summed with a modified photomodification factor (r2 = 0.92). The greatest correlation was observed with median effective concentration data. This research provides evidence that models incorporating factors for photosensitization and photomodification have interspecies applicability.

Opinion: Taking phytoremediation from proven technology to accepted practice.

Phytoremediation is the use of plants to extract, immobilize, contain and/or degrade contaminants from soil, water or air. It can be an effective strategy for on site and/or in situ removal of various contaminants from soils, including petroleum hydrocarbons (PHC), polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), solvents (e.g., trichloroethylene [TCE]), munitions waste (e.g., 2,4,6-trinitrotoluene [TNT]), metal(loid)s, salt (NaCl) and radioisotopes. Commercial phytoremediation technologies appear to be underutilized globally. The primary objective of this opinion piece is to discuss how to take phytoremediation from a proven technology to an accepted practice. An overview of phytoremediation of soil is provided, with the focus on field applications, to provide a frame of reference for the subsequent discussion on better utilization of phytoremediation. We consider reasons why phytoremediation is underutilized, despite clear evidence that, under many conditions, it can be applied quite successfully in the field. We offer suggestions on how to gain greater acceptance for phytoremediation by industry and government. A new paradigm of phytomanagement, with a specific focus on using phytoremediation as a "gentle remediation option" (GRO) within a broader, long-term management strategy, is also discussed.

Photoinduced toxicity of polycyclic aromatic hydrocarbons to Daphnia magna: ultraviolet-mediated effects and the toxicity of polycyclic aromatic hydrocarbon photoproducts.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants known for their photoinduced toxicity. This toxicity may occur through two mechanisms: Photosensitization, and photomodification. Photosensitization generally leads to the production of singlet oxygen, a reactive oxygen species that is highly damaging to biological molecules. Photomodification of PAHs, usually via oxidation, results in the formation of new compounds and can occur under environmentally relevant levels of actinic radiation. The toxicities of 16 intact PAHs to Daphnia magna were assessed under two ultraviolet radiation conditions. The toxicity of intact PAHs generally increased in the presence of full-spectrum simulated solar radiation relative to that in the presence of visible light plus ultraviolet A only. Despite the knowledge of a bipartite mechanism of phototoxicity that includes photosensitization and photomodification, few studies have examined the effects of PAH photoproducts on animals. To expand the existing data, 14 PAH photoproducts (oxy-PAHs) also were assayed, most of which were highly toxic without further photomodification. Two photoproducts of benzo[a]pyrene, 1,6- and 3,6-benzo[a]pyrenequinone, were the most toxic compounds tested, followed closely by benz[a]anthraquinone. Each of these three compounds had a median effective concentration in the low nanomolar range. The data presented highlight the effects of ultraviolet radiation on mediating PAH toxicity and the need to analyze absorption spectra of contaminants in the prediction of photoinduced toxicity. The importance of the role of photomodification also is stressed, because several oxy-PAHs, an unregulated group of contaminants, were highly toxic to D. magna, a key bioindicator species in aquatic ecosystems.

Assessment of the toxicity of mixtures of copper, 9,10-phenanthrenequinone, and phenanthrene to Daphnia magna: evidence for a reactive oxygen mechanism.

Polycyclic aromatic hydrocarbons and their derivatives are ubiquitous environmental contaminants. They are commonly present in complex mixtures with other contaminants, such as metals. The toxicities of phenanthrene (PHE) and 9,10-phenanthrenequinone (PHQ) with or without Cu were determined using Daphnia magna. Copper was the most toxic among the three chemicals tested, followed by PHQ and then PHE, with 48-h median effective concentrations (EC50s) of 0.96, 1.72, and 5.33 microM, respectively. Copper at 0.31 microM, or approximately the 5% effective concentration, decreased the EC50 of PHQ from 1.72 to 0.28 microM. Likewise, PHQ at 1.2 microM, or approximately the 10% effective concentration, significantly lowered the EC50 of Cu from 0.96 to 0.30 microM. This synergistic effect was not observed, however, in mixtures of Cu and PHE based on the response addition model. Assimilation of Cu wasfound to be similar with or without PHQ at increasing external concentrations of Cu, indicating that the increased toxicity of their mixtures is physiologically based. The ability of Cu plus PHQ to generate reactive oxygen species (ROS) was measured as well. Copper alone caused elevated ROS levels at a low concentration (0.63 microM). With PHQ present, however, this elevation in ROS occurred at an even lower Cu level (0.31 microM). Possible attenuation effects of ascorbic acid (vitamin C) on toxicity and ROS production induced by Cu, PHQ, and their mixtures were then examined. Ascorbic acid protected against Cu and Cu-plus-PHQ mixture-mediated toxicity but did not affect PHQ toxicity. Ascorbic acid also lowered ROS levels in the presence of Cu and Cu plus PHQ. We conclude that there exist potential toxic interactions between metals and modified PAHs and that these interactions can involve ROS formation.

Ultraviolet wavelength dependence of photomorphological and photosynthetic responses in Brassica napus and Arabidopsis thaliana.

Among the photomorphological responses in plants induced by ultraviolet-B radiation (UVB; 290 nm-320 nm) are leaf asymmetry, leaf thickening and cotyledon curling. We constructed an action spectrum of cotyledon curling in light-grown Brassica napus to characterize the UVB photoreceptor that initiates this response. Cotyledon curling was also characterized in Arabidopsis thaliana. Peak efficiency for this response occurred between 285 and 290 nm. Additionally, UVB-induced changes in epidermal cells from A. thaliana cotyledons were assessed because they are the likely site of UVB photoreception that leads to curling. Investigation of cellular structure, chlorophyll a fluorescence and chlorophyll concentration indicated that cotyledon curling is not concomitant with gross cellular damage or inhibition of photosynthesis, which only occurred in response to wavelengths <280 nm. Many UVB effects are apparently an indirect consequence of UVB radiation, dependent on UVB-mediated increases in reactive oxygen species (ROS) that either act as a signal in the UVB transduction pathway or cause oxidative damage. The cotyledon curling response was impeded by ascorbate and cystine, ROS scavengers and was promoted by H(2)O(2), a ROS. We suggest that following absorption by a UVB chromophore, ROS are generated via photosensitization, ultimately leading to cotyledon curling.

Tolerance of transgenic canola expressing 1-aminocyclopropane-1-carboxylic acid deaminase to growth inhibition by nickel.

Plant growth-promoting bacteria are useful to phytoremediation strategies in that they confer advantages to plants in contaminated soil. When plant growth-promoting bacteria contain the enzyme 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase, the bacterial cell acts as a sink for ACC, the immediate biosynthetic precursor of the plant growth regulator ethylene thereby lowering plant ethylene levels and decreasing the negative effects of various environmental stresses. In an effort to gain the advantages provided by bacterial ACC deaminase in the phytoremediation of metals from the environment two transgenic canola lines with the gene for this enzyme were generated and tested. In these transgenic canola plants, expression of the ACC deaminase gene is driven by either tandem constitutive cauliflower mosaic virus (CaMV) 35S promoters or the root specific rolD promoter from Agrobacterium rhizogenes. Following the growth of transgenic and non-transformed canola in nickel contaminated soil, it was observed that the rolD plants demonstrate significantly increased tolerance to nickel compared to the non-transformed control plants.

Biochemical responses of the aquatic higher plant Lemna gibba to a mixture of copper and 1,2-dihydroxyanthraquinone: synergistic toxicity via reactive oxygen species.

Metals and polycyclic aromatic hydrocarbons (PAHs) are known to be toxic to plants. Because metals and PAHs often are cocontaminants in the environment, plants can be subjected to damage caused by their combined effects. We recently found that copper and an oxygenated PAH (1,2-dihydroxyanthraquinone [1,2-dhATQ]) synergistically are toxic to plants. This synergistic toxicity was linked indirectly to production of reactive oxygen species (ROS). In this study, plant growth, chlorophyll pigments, protein accumulation, and ROS production were chosen as endpoints to assess the mechanism of toxicity of copper and 1,2-dhATQ to Lemna gibba in more detail. Because copper and PAHs can generate ROS, we assayed for specific antioxidant enzymes: Superoxide dismutase (SOD), glutathione reductase (GR), and ascorbate peroxidase (APX). Copper treatment at a concentration that did not cause growth inhibition resulted in upregulation of Mn SOD, Cu-Zn SOD, and APX. At a level that moderately was toxic to plants, 1,2-dhATQ did not alter significantly the levels of these antioxidant enzymes. However, a synergistically toxic mixture of copper plus 1,2-dhATQ upregulated Cu-Zn SOD, Mn SOD, and GR, although APX activity was downregulated. When plants were treated with the ROS scavenger dimethyl thiourea (DMTU), enhanced toxicity and formation of ROS caused by the mixture both were diminished substantially. However, 1,2-dhATQ toxicity was not affected significantly by DMTU. Based on this study, the toxicity caused by the mixture of copper plus 1,2-dhATQ directly can be connected to elevated levels of ROS.

Identification of six differentially expressed genes in response to copper exposure in the aquatic plant Lemna gibba (Duckweed).

Aquatic plants are susceptible to metal pollution and provide an entry point for metals, such as copper, into the aquatic biosphere. Exposure of the aquatic plant Lemna gibba to copper has been associated with the production of reactive oxygen species (ROS) and oxidative damage, caused in large part by the ability of this metal to redox cycle. In particular, copper-mediated production of ROS, a known group of signaling molecules, triggers numerous defense responses in L. gibba. Therefore, the objective of the present study was to examine to what extent acute copper exposure alters gene expression. First, the kinetics of copper uptake was assessed to determine if assimilation occurred within the short exposures needed to induce gene expression. Subsequently, using differential display polymerase chain reaction, we identified six genes with expressions that were putatively altered in response to copper. Differential expression was confirmed by northern hybridization analysis and showed that copper causes an accumulation of transcripts that encode for callose synthase, heat shock protein 90, serine decarboxylase, and the biotin carboxylase subunit of acetyl-coenzyme A carboxylase. Conversely, copper caused a decline in transcript levels for genes encoding the HAP5 subunit of the heme-activated protein (HAP) transcription factor in addition to the chloroplast nucleoid DNA-binding protein CND41. Interestingly, the expressions of these genes are sensitive to cellular ROS levels. We believe that these gene products provide valuable information regarding the molecular mechanisms of copper toxicity.