Copper Toxicity Research Articles

Toxicological effects of copper on bioaccumulation and mRNA expression of antioxidant, immune, and apoptosis-related genes in Chinese striped-necked turtle (Mauremys sinensis).

Heavy metals are among the most ubiquitous environmental pollutants of recent decades. Copper is commonly used to control algal blooms or macrophyte and waste infestations, its ambient concentration has increased significantly, indicating possible environmental risk. To investigate the effects of copper exposure on bioaccumulation, antioxidant defense, immune response, and apoptosis in the Chinese Striped-necked Turtle Mauremys sinensis, three experimental groups, control (0.0mg/L), Cu2 (2mg/L) and Cu4 (4mg/L) were designed, and sampled at 14 and 28days. Results showed that copper accumulates in different organs depending on the concentration and exposure time, Liver > Kidney > Gut > Heart > Brain > Muscle and the time order was 28days > 14days. The liver enzymes AST, ALT, and ALP decreased when the turtles were exposed to copper stress, while the contents of bilirubin TBIL, DBIL, IBIL, and LDH showed a significant upward trend. Similarly, the mRNA expression level of acetylcholinesterase AChE in the brain was significantly downregulated upon copper exposure. An upward trend was noticed in the liver Metallothionein MT mRNA expression levels compared to the control group. The mRNA expression levels of antioxidant enzymes CAT, SOD, MnSOD, and GSH-PX1 in the liver increased initially and then significantly decreased. Furthermore, the relative mRNA expression levels of inflammatory cytokines IL-1β, IL-8, TNF-α, and IFN-γ involved in inflammatory response significantly upregulated. Copper significantly increased the hepatic mRNA transcription of heat shock proteins HSP70 and HSP90 at different exposure durations. In addition, the relative mRNA levels of caspase3, caspase8, and caspase9 related to the caspase-dependent apoptotic pathway significantly increased under copper stress. These results explain that copper toxicity causes bioaccumulation, promotes oxidative stress, obstructs immunity, and induces inflammation and apoptosis by altering their gene expression levels in M. sinensis.

Genome editing without nucleases confers proliferative advantage to edited hepatocytes and corrects Wilson disease.

Application of classic liver-directed gene replacement strategies is limited in genetic diseases characterized by liver injury due to hepatocyte proliferation resulting in decline of therapeutic transgene expression and potential genotoxic risk. Wilson disease (WD) is a life-threating autosomal disorder of copper homeostasis caused by pathogenic variants in copper transporter ATP7B and characterized by toxic copper accumulation, resulting in severe liver and brain diseases. Genome editing holds promise for the treatment of WD, nevertheless to rescue copper homeostasis ATP7B function must be restored in at least 25% of the hepatocytes, which surpasses by far genome editing correction rates. We applied a liver-directed nuclease-free genome editing approach, based on adeno-associated viral vector (AAV) -mediated targeted integration of a promoterless mini-ATP7B cDNA into the Alb locus. Administration of AAV-Alb-mini-ATP7B in two WD mouse models resulted in extensive liver repopulation by genome edited hepatocytes holding a proliferative advantage over non-edited ones, and ameliorated liver injury and copper metabolism. Furthermore, combination of genome editing with a copper chelator currently used for WD treatment, achieved greater disease improvement compared to chelation therapy alone. Nuclease-free genome editing provided therapeutic efficacy and may represent a safer and longer lasting alternative to classic gene replacement strategies for WD.

Copper drives prion protein phase separation and modulates aggregation.

Prion diseases are characterized by prion protein (PrP) transmissible aggregation and neurodegeneration, which has been linked to oxidative stress. The physiological function of PrP seems related to sequestering of redox-active Cu2+, and Cu2+ dyshomeostasis is observed in prion disease brain. It is unclear whether Cu2+ contributes to PrP aggregation, recently shown to be mediated by PrP condensation. This study indicates that Cu2+ promotes PrP condensation in live cells at the cell surface and in vitro through copartitioning. Molecularly, Cu2+ inhibited PrP β-structure and hydrophobic residues exposure. Oxidation, induced by H2O2, triggered liquid-to-solid transition of PrP:Cu2+ condensates and promoted amyloid-like PrP aggregation. In cells, overexpression of PrPC initially protected against Cu2+ cytotoxicity but led to PrPC aggregation upon extended copper exposure. Our data suggest that PrP condensates function as a buffer for copper that prevents copper toxicity but can transition into PrP aggregation at prolonged oxidative stress.

Zinc's Role in Mitigating Copper Toxicity for Plants and Microorganisms in Industrially Contaminated Soils: A Review

This review focuses on the issue of metal antagonism in soils contaminated by multiple metals as a result of industrial emissions. Building upon previous findings in aquatic ecosystems, the potential of zinc to mitigate copper toxicity in more complex soil systems is explored. A range of studies investigating the role of zinc in reducing copper toxicity to plants and microorganisms in soils contaminated by copper mining in central Chile are examined. The mechanisms underlying metal interactions in soils, including the terrestrial biotic ligand model and the intensity/capacity/quantity concept, are thoroughly discussed. Furthermore, the review underscores the pressing need for future studies to enhance our understanding and develop effective strategies for mitigating copper toxicity in industrially contaminated soils.

Copper bioconcentration and bioaccumulation in different eucalyptus species and their impact on plant physiology

Some plant species can bioaccumulate copper in their tissues and can be used for phytoremediation in contaminated soils. This work aimed to determine the bioconcentration and bioaccumulation of copper in eucalyptus species and how they affect the physiology of plants. The experimental design was completely randomized in a factorial arrangement (4 × 6), with four species of eucalyptus (Eucalyptus grandis W. Hill ex. Maiden, Corymbia citriodora (Hook.) K.D. Hill and L.A.S. Johnson, Eucalyptus saligna Smith and Eucalyptus dunnii Maiden), six doses of copper (zero, 80, 160, 240, 320, 400 mg kg−1), with twelve repetitions, carried out for 120 days in a greenhouse condition. The exotic species showed a high potential for growing in soils with high copper concentrations and also a potential for copper bioremediation. The highest copper concentration accumulation was shown mainly in the root system of the seedlings of Eucalyptus grandis, Eucalyptus saligna, Eucalyptus dunnii and Corymbia citriodora. E. saligna the copper toxicity was less pronounced, and the plants showed better physiologic effects. Cu levels increased with the doses, with greater intensity for species E. grandis and E. saligna with values of 21.1 and 15.5 mg kg−1, respectively. Also, the species showed a high capacity for copper bioaccumulation in the root system and showed high characteristics for phytoaccumulation and phytoextraction. So, these plants have great characteristics to grow in copper-contaminated sites being useful to reduce free copper in the environment.

Assessment of sediments contamination by potential toxic elements, in relationships with the physicochemical characteristics of Moroccan Oued Fez river

The Oued Fez river has long been significantly impacted by various sources of pollution, including industrial activities. However, there is limited available data concerning the presence of potentially harmful toxic elements that could affect human health in this area. The primary aim of this study was to evaluate the spatial and temporal distribution of nine Potential Toxic Elements (PTEs) Aluminum (Al), Cadmium (Cd), Chromium (Cr), Copper (Cu), Iron (Fe), Lead (Pb), Nickel (Ni), Arsenic (As), and Zinc (Zn) in the sediment of the Oued Fez river. In both February and August, we collected four sediment samples at six different locations along the Oued Fez. To ensure sample consistency at each station, we mixed the sediment samples, dried them, and finely ground them into a powder. We then used acid etching to dissolve the samples, and the concentrations of these PTEs were determined using the ICP-AES method. The findings revealed that heavy metal concentrations were higher in February compared to August, with a gradual increase observed from upstream to downstream areas of the river. Specifically, in February, site six (S6) exhibited the highest values for Fe at 27,749.6 mg.kg−1 and Al at 13,967.2 mg.kg−1. Additionally, the PCA analysis and assessment of sediment contamination indicated that the sediment was more contaminated with Fe, Al, and Zn, especially at upstream stations (S5 and S6). This contamination can be attributed to their proximity to the industrial area. These results clearly demonstrate that anthropogenic factors, particularly industrial activities, are contributing to increased sediment contamination by PTEs. This contamination not only jeopardizes the environmental integrity of this site but also poses a significant threat to human health.

A new Caenorhabditis elegans model to study copper toxicity in Wilson disease.

Wilson disease (WD) is caused by mutations in the ATP7B gene that encodes a copper (Cu) transporting ATPase whose trafficking from the Golgi to endo-lysosomal compartments drives sequestration of excess Cu and its further excretion from hepatocytes into the bile. Loss of ATP7B function leads to toxic Cu overload in the liver and subsequently in the brain, causing fatal hepatic and neurological abnormalities. The limitations of existing WD therapies call for the development of new therapeutic approaches, which require an amenable animal model system for screening and validation of drugs and molecular targets. To achieve this objective, we generated a mutant Caenorhabditis elegans strain with a substitution of a conserved histidine (H828Q) in the ATP7B ortholog cua-1 corresponding to the most common ATP7B variant (H1069Q) that causes WD. cua-1 mutant animals exhibited very poor resistance to Cu compared to the wild-type strain. This manifested in a strong delay in larval development, a shorter lifespan, impaired motility, oxidative stress pathway activation, and mitochondrial damage. In addition, morphological analysis revealed several neuronal abnormalities in cua-1 mutant animals exposed to Cu. Further investigation suggested that mutant CUA-1 is retained and degraded in the endoplasmic reticulum, similarly to human ATP7B-H1069Q. As a consequence, the mutant protein does not allow animals to counteract Cu toxicity. Notably, pharmacological correctors of ATP7B-H1069Q reduced Cu toxicity in cua-1 mutants indicating that similar pathogenic molecular pathways might be activated by the H/Q substitution and, therefore, targeted for rescue of ATP7B/CUA-1 function. Taken together, our findings suggest that the newly generated cua-1 mutant strain represents an excellent model for Cu toxicity studies in WD.

Mechanisms of Copper Toxicity and Tolerance in the Aquatic Moss Taxiphyllum barbieri.

Aquatic habitats are very frequently polluted with different kinds of xenobiotics, including heavy metals. For biomonitoring studies of aquatic pollution, algae are frequently used, as they do not contain protective cuticle on the surface of their thalli and can accumulate pollutants over the whole surface of thalli. However, this is a feature of most cryptogams. For this reason, we assessed the sensitivity of the aquatic moss Taxiphyllum barbieri (Java moss) to copper excess in a short-term study. Moss T. barbieri belongs to the common aquatic plants originating from Southeast Asia. To test the sensitivity (or tolerance) of the moss to excess Cu, selected concentrations (50, 250 and 500 µM) were employed in our 24 h studies. Total and intracellular Cu accumulation positively correlated with Cu availability in the water. This total and intracellular Cu accumulation was negatively correlated with decreased intracellular K content. Excess Cu negatively affected the composition of assimilation pigments and soluble proteins. Cu caused increased peroxidation of membrane lipids assessed using TBARS assay. Excess Cu decreased GSH to GSSG ratio and ascorbic acid content. We did not observe phytochelatin synthesis in this moss. The roles of selected amino acids, their intermediates and derivatives, as well as S-containing nucleosides and phenolic acids in Cu homeostasis and toxicity or tolerance were evaluated. We assume that this moss has potential for future employment in water quality evaluation.

Protective role of fullerenol and arginine C60 fullerene against copper toxicity in cucumber

Copper (Cu), when in excess, is one of the most toxic and hazardous metals to all living organisms, including plants. Engineered nanomaterials have the potential for increasing crop protection. However, the protective role of fullerenes (carbon-based nanoparticles with wide application in various areas) against Cu toxicity in plants is, so far, understudied. The present study investigated whether fullerenes can potentially alleviate Cu toxicity in plants (Cucumis sativus L.). Two water-soluble fullerene C60 derivatives were examined: fullerenol [C60(OH)22–24] and arginine-functionalized fullerene [C60(C6H13N4O2)8H8], under controlled conditions using hydroponics. Plants treated with 15 μM of Cu exhibited typical symptoms of Cu toxicity: impaired growth, leaf chlorosis, reduced photosynthetic activity, nutritional imbalances, and enhanced lipid peroxidation. These symptoms were alleviated in the presence of fullerene derivatives with arginine C60 having the more pronounced effect. Improved cucumber Cu tolerance was attributable to Cu buffering in the root zone (roots and medium), which caused a dramatic decline in Cu transport towards leaves and the elimination of oxidative damage. The Cu removal efficacy of arginine C60 was much greater than that of fullerenol. These fullerenes acted in a dose-dependent manner and removed Cu selectively without significant modification of the bioavailability of other essential nutrients. Treatment with free arginine did not affect Cu immobilization or Cu toxicity. These results suggest that the surface chemistry of the fullerene core is important for the protection of plants under excessive Cu conditions. The information offered a new approach to preparing promising practical materials for alleviating Cu toxicity in plants with potential application in fields.

Alterations in plasma and erythrocyte membrane fatty acid composition following exposure to toxic copper level affect membrane deformability and fluidity in female wistar rats

BackgroundDeformability and fluidity function of the red blood cell membrane are properties defined by the lipid composition. Toxic copper level induces membrane lipid peroxidation which could cause membrane instability. This study therefore investigated the effect of exposure to toxic copper level for 30 days on red blood cell membrane deformability and fluidity in female Wistar rats. MethodsTwelve (12) female Wistar rats (160 ± 10 g) were randomly grouped (n = 6) into control (given 0.1 ml distilled water p.o.) and copper-toxic (100 mg/kg Copper Sulphate, p.o.), and treated for 30 days. Plasma obtained and RBC membrane prepared from blood collected over EDTA post-treatment were assayed for total cholesterol (TC), phospholipids and fatty acid profile using spectrophotometry and Gas chromatography while heparinized blood was subjected to fragility test. Data were analyzed using student T-test for statistical significance at p < 0.05. Results and conclusionPlasma TC increased by 4.33% while RBC membrane TC decreased by 20.32% in copper-toxic group compared to control. Compared to control, excess copper significantly increased membrane phospholipids level (0.72 ± 0.01 vs 0.59 ± 0.04 mg/dL) but reduced membrane cholesterol/phospholipid ratio (46.61 ± 4.72 vs 72.66 ± 6.47) and stability (by 23.53%). Number of cis- and saturated fatty acids increased in copper-treated plasma and RBC membrane compared to control. Exposure to toxic copper level alters erythrocyte membrane fluidity and deformability by disrupting membrane lipid composition, saturation, bond configuration in phospholipids and permeability.

Changes in Isoflavonoid and Flavonoid Content in Soybean Leaves Affected by UV-B or Copper

Abstract The production of antioxidant flavonoids is an important aspect of plant defence mechanisms under stress conditions. The hypothesis of increased antioxidant flavonol quercetin and isoflavone content in soybean leaves subjected to UV-B irradiation and copper toxicity was proposed, and their accumulation was investigated using HPLC analysis. However, the accumulation of antioxidant quercetin was not induced by either UV-B irradiation or copper treatment. UV-B irradiation strongly induced isoflavone aglycones, mainly genistein, daidzein, formononetin, and to a lesser extent, biochanin A. Isoflavone glycosides, genistin and daidzin were also induced. Copper treatment, whether applied to leaves or root media, resulted in the accumulation of genistin and its malonylated form, whereas aglycones were only slightly affected. The higher accumulation of genistein over daidzein after copper treatment could be related to the higher chelating ability of genistein and its glycosides. Soybean isoflavones, which possess antioxidant and chelating properties, rather than flavonols, play a crucial role in the plant‘s defence mechanisms against both UV-B irradiation and copper toxicity.

MeGLYI-13, a Glyoxalase I Gene in Cassava, Enhances the Tolerance of Yeast and Arabidopsis to Zinc and Copper Stresses.

Although zinc and copper are the two essential nutrients necessary for plant growth, their excessive accumulation in soil not only causes environmental pollution but also seriously threatens human health and inhibits plant growth. The breeding of plants with novel zinc or copper toxicity tolerance capacities represents one strategy to address this problem. Glyoxalase I (GLYI) family genes have previously been suggested to be involved in the resistance to a wide range of abiotic stresses, including those invoked by heavy metals. Here, a MeGLYI-13 gene cloned from a cassava SC8 cultivar was characterized with regard to its potential ability in resistance to zinc or copper stresses. Sequence alignment indicated that MeGLYI-13 exhibits sequence differences between genotypes. Transient expression analysis revealed the nuclear localization of MeGLYI-13. A nuclear localization signal (NLS) was found in its C-terminal region. There are 12 Zn2+ binding sites and 14 Cu2+ binding sites predicted by the MIB tool, of which six binding sites were shared by Zn2+ and Cu2+. The overexpression of MeGLYI-13 enhanced both the zinc and copper toxicity tolerances of transformed yeast cells and Arabidopsis seedlings. Taken together, our study shows the ability of the MeGLYI-13 gene to resist zinc and copper toxicity, which provides genetic resources for the future breeding of plants resistant to zinc and copper and potentially other heavy metals.

Mitochondrial Oxidative Stress Is the General Reason for Apoptosis Induced by Different-Valence Heavy Metals in Cells and Mitochondria.

This review analyzes the causes and consequences of apoptosis resulting from oxidative stress that occurs in mitochondria and cells exposed to the toxic effects of different-valence heavy metals (Ag+, Tl+, Hg2+, Cd2+, Pb2+, Al3+, Ga3+, In3+, As3+, Sb3+, Cr6+, and U6+). The problems of the relationship between the integration of these toxic metals into molecular mechanisms with the subsequent development of pathophysiological processes and the appearance of diseases caused by the accumulation of these metals in the body are also addressed in this review. Such apoptosis is characterized by a reduction in cell viability, the activation of caspase-3 and caspase-9, the expression of pro-apoptotic genes (Bax and Bcl-2), and the activation of protein kinases (ERK, JNK, p53, and p38) by mitogens. Moreover, the oxidative stress manifests as the mitochondrial permeability transition pore (MPTP) opening, mitochondrial swelling, an increase in the production of reactive oxygen species (ROS) and H2O2, lipid peroxidation, cytochrome c release, a decline in the inner mitochondrial membrane potential (ΔΨmito), a decrease in ATP synthesis, and reduced glutathione and oxygen consumption as well as cytoplasm and matrix calcium overload due to Ca2+ release from the endoplasmic reticulum (ER). The apoptosis and respiratory dysfunction induced by these metals are discussed regarding their interaction with cellular and mitochondrial thiol groups and Fe2+ metabolism disturbance. Similarities and differences in the toxic effects of Tl+ from those of other heavy metals under review are discussed. Similarities may be due to the increase in the cytoplasmic calcium concentration induced by Tl+ and these metals. One difference discussed is the failure to decrease Tl+ toxicity through metallothionein-dependent mechanisms. Another difference could be the decrease in reduced glutathione in the matrix due to the reversible oxidation of Tl+ to Tl3+ near the centers of ROS generation in the respiratory chain. The latter may explain why thallium toxicity to humans turned out to be higher than the toxicity of mercury, lead, cadmium, copper, and zinc.

Identification of a novel transcriptional regulator, CorR, for copper stress response in Edwardsiella piscicida.

Copper plays a vital role in the host-pathogen interface, potentially making components of the bacterial copper response suitable targets for the development of innovative antimicrobial strategies. The anti-copper arsenal of intracellular pathogens has expanded as an adaptation to survive copper toxicity in order to escape intracellular killing by the host immune system. Herein, we employed transposon insertion sequencing to investigate the genetic mechanisms underlying the survival of Edwardsiella piscicida under copper stress. A novel transcriptional regulator, ETAE_2324 (named CorR), was identified to participate in the response to copper ions by controlling the expression of copA, the core component of cytoplasmic copper homeostasis. Furthermore, CorR regulated the expression of virulent determinant eseB, influencing the in vivo colonization of E. piscicida. Collectively, our results contribute to the comprehension of the underlying mechanism of the adaption of intracellular pathogens to copper stress during bacterial infections. IMPORTANCE Copper ions play a pivotal role in the interaction between bacteria and the host during infection. The host's innate immune system employs copper ions for their bactericidal properties, thereby making bacterial copper tolerance a crucial determinant of virulence. Edwardsiella piscicida, a significant marine pathogen, has caused substantial losses in the global aquaculture industry. To comprehensively investigate how E. piscicida responds to copper stress, we utilized transposon insertion sequencing to explore genes associated with copper tolerance in culture media containing different concentrations of copper ions. A novel transcriptional regulator, CorR, was identified to respond to copper ions and regulates the expression of crucial components of copper homeostasis CopA, along with the essential virulence factor EseB. These findings offer valuable insights into the underlying mechanisms that govern bacterial copper tolerance and present novel perspectives for the development of vaccines and therapeutic strategies targeting E. piscicida.

ATP7B R778L mutant hepatocytes resist copper toxicity by activating autophagy and inhibiting necroptosis

Wilson’s disease (WD) is an inherited disease characterized by copper metabolism disorder caused by mutations in the adenosine triphosphatase copper transporting β gene (ATP7B). Currently, WD cell and animal model targeting the most common R778L mutation in Asia is lacking. In addition, the mechanisms by which hepatocytes resist copper toxicity remain to be further elucidated. In this study, we aimed to construct a novel WD cell model with R778L mutation and dissected the molecular basics of copper resistance. A novel HepG2 cell line stably expressing the ATP7B R778L gene (R778L cell) was constructed. The expression of necroptosis- and autophagy-related molecules was detected by PCR and Western blot (WB) in wild-type (WT) HepG2 and R778L cells with or without CuSO4 treatment. In addition, we detected and compared the levels of autophagy and necroptosis in CuSO4-treated R778L cells with the activation and inhibition of autophagy. Moreover, the mRNA and protein levels of autophagy and necroptosis signaling molecules were compared in R778L cells with the overexpression and knockdown of Unc-51 Like Autophagy Activating Kinase 1 (ULK1) and Autophagy Related 16 Like 1 (ATG16L1). We successfully constructed an R778L mutation HepG2 cell line. CuSO4 triggered the enhanced expression of autophagy and necroptosis signaling molecules in WT HepG2 cells and R778L cells. Remarkably, higher levels of autophagy and necroptosis were observed in R778L cells compared with those in WT cells. Autophagy activation led to weakened necroptosis mediated by RIPK3 and MLKL, conversely, autophagy inhibition brought about enhanced necroptosis. At the molecular level, ULK1- and ATG16L1 overexpression resulted in reduced necroptosis levels and vice versa. ULK1- and ATG16L1-mediated autophagy activation protects hepatocytes against RIPK3- and MLKL-mediated necroptosis in our new WD cell model treated with CuSO4. Targeted therapy by autophagy activation or necroptosis inhibition may be a novel and effective strategy to treat WD.

Effects of Inorganic and Organic Pollutants on the Biomarkers' Response of Cerastoderma edule under Temperature Scenarios.

Currently, there is increased chemical pollution, and climate change is a major concern to scientific, political and social communities globally. Marine systems are very susceptible to changes, and considering the ecological and economic roles of bivalve species, like Cerastoderma edule, studies evaluating the effects of both stressors are of great importance. This study intends to (a) determine the toxicity of copper (Cu) and oxyfluorfen at the lethal level, considering the temperature; (b) assess the changes in the antioxidant defence enzymes as a consequence of the simultaneous exposure to chemical and warming pressures; and (c) determine if lipid peroxidation (LPO) and neurotoxic effects occur after the exposure to chemical and temperature stressors. C. edule was exposed to Cu and oxyfluorfen at different temperatures (15 °C, 20 °C and 25 °C) for 96 h. The ecotoxicological results reveal a higher tolerance of C. edule to oxyfluorfen than to Cu, regardless of the temperature. The antioxidant defence system revealed efficiency in fighting the chemicals' action, with no significant changes in the thiobarbituric reactive species (TBARS) levels to 15 °C and 20 °C. However, a significant inhibition of acetylcholinesterase (AChE) was observed on the organisms exposed to oxyfluorfen at 20 °C, as well as a decreasing trend on the ones exposed to Cu at this temperature. Moreover, the catalase (CAT) showed a significant increase in the organisms exposed to the two highest concentrations of Cu at 15 °C and in the ones exposed to the highest concentration of oxyfluorfen at 20 °C. Looking at the temperature as a single stressor, the organisms exposed to 25 °C revealed a significant increase in the TBARS level, suggesting potential LPO and explaining the great mortality at this condition.

Metallothionein: a game changer in histopathological diagnosis of Wilson disease.

Wilson disease (WD) is a genetic disorder of copper metabolism caused by mutations in the ATP7B gene. Toxic copper accumulation leads to hepatic, neurologic, and psychiatric disorders with variable presentation. Metallothionein (MT) immunohistochemistry was proposed as a diagnostic marker. MT immunohistochemistry was performed on liver specimens of WD patients (n = 64) and control cases (n = 160) including acute liver failure, steatotic liver disease, autoimmune hepatitis, normal liver, primary biliary cholangitis, primary and secondary sclerosing cholangitis, and progressive familial intrahepatic cholestasis. The optimal cutoff for detection of WD was determined by receiver operating characteristic (ROC) analysis. At least moderate staining in >50% of hepatocytes was observed in 81% of analysed liver specimens (n = 56/69) of WD patients, while only five control cases showed this staining pattern. The sensitivity, specificity, and accuracy for a new diagnosis of WD were 85.7%, 96.9%, and 94.9%, respectively. Sensitivity in nonfibrotic patients was 70.6% and this MT pattern was robust in small biopsies. Thehepatic copper concentration was similar between MT-positive and MT-negative liver samples (P > 0.05). Zinc treatment may induce hepatocellular MT expression. Kayser-Fleischer rings (50% versus 15%) and neurologic disorders (50% versus 13%) were significantly more prevalent in MT-negative compared to MT-positive WD patients, respectively. MT immunostaining is an excellent biomarker for histological diagnosis of WD, should be incorporated in the diagnostic work-up of patients with potential WD, and is useful in a modified Leipzig score.

Quercetin Alleviates Toxicity Induced by High Levels of Copper in Porcine Follicular Granulosa Cells by Scavenging Reactive Oxygen Species and Improving Mitochondrial Function.

CuSO4 is the most commonly used feed additive in pig production at present, but long-term ingestion of excessive copper would lead to chronic copper toxicity. High copper could reduce the reproductive efficiency of sows and seriously affect the development of the pig industry. Quercetin (QUE), a powerful antioxidant, reduces toxicity of a number of heavy metals. Porcine granulosa cells (pGCs) are crucial to the fate of follicle development. The present study found that high concentrations of CuSO4 induced ROS production, which resulted in decreased mRNA expression of antioxidant-related genes GPX4, CAT, and SOD2 and increased mRNA expression of SOD1, TRX, and HO-1. The protein expression of antioxidant enzymes SOD2 and HO-1 decreased. Moreover, the concentration of MDA increased, the activity of CAT decreased, and the content of GSH decreased. After high copper treatment, the mitochondrial membrane potential (MMP) was decreased and the morphological structure was changed. However, the combined treatment with Quercetin (QUE) reversed these changes, and the level of cellular oxidative stress decreased. Therefore, we conclude that high copper has oxidative toxicity to pGCs, and QUE could remove the ROS induced by high copper, protect mitochondria from oxidative stress damage, and improve the function of pGCs.

Acute Toxicity of Copper to Three Species of Pacific Salmon Fry in Water with Low Hardness and Low Dissolved Organic Carbon.

Proposed development of a mine within Alaska's Bristol Bay watershed (USA) has raised concerns about the potential impact of copper (Cu) on Pacific salmon (Oncorhynchus spp.). We conducted 96-h flow-through bioassays using low-hardness and low dissolved organic carbon water to determine the acute lethal toxicity of Cu to sockeye (Oncorhynchus nerka), Chinook (Oncorhynchus tshawytscha), and coho salmon (Oncorhynchus kisutch) fry. We aimed to determine Cu toxicity under field-relevant water quality conditions and to assess three methods of calculating ambient Cu criteria: the biotic ligand model (BLM), a multiple linear regression model endorsed by the US Environmental Protection Agency, and the hardness-based model currently used by the State of Alaska. The criteria generated by all models were below 20% lethal Cu concentrations by factors ranging from 2.2 to 54.3, indicating that all criteria would be protective against mortality. The multiple linear regression-based criteria were the most conservative and were comparable to BLM-based criteria. The median lethal concentrations (LC50s) for sockeye, Chinook, and coho were 35.2, 23.9, and 6.3 µg Cu/L, respectively. We also used the BLM to predict LC50s for each species. Model predictions differed from empirical LC50s by factors of 0.7 for sockeye and Chinook salmon, and 1.1 for coho salmon. These differences fell within the acceptable range of ±2, indicating the model's accuracy. We calculated critical lethal Cu accumulation values for each species to account for differing water chemistry in each bioassay; the present study revealed that coho salmon were most sensitive to Cu, followed by sockeye and Chinook salmon. Our findings underscore the importance of considering site- and species-specific factors when modeling Cu toxicity. The empirical data we present may enhance Cu risk assessments for Pacific salmon. Environ Toxicol Chem 2023;42:2440-2452. © 2023 SETAC.

Increased ascorbic acid synthesis by overexpression of AcGGP3 ameliorates copper toxicity in kiwifruit

The widespread application of copper (Cu) -based fertilizers and pesticides could increase the accumulation of Cu in kiwifruit. According to a global survey, red- and yellow-fleshed kiwifruit contained more elevated amounts of Cu than green-fleshed kiwifruit due to weaker disease resistance and higher use of Cu pesticides. Intriguingly, our research revealed that external and endogenous ascorbic acid (AsA) reduced the phenotypic and physiological injury of Cu toxicity in kiwifruit. Cu stress assays and transcriptional analysis have shown that Cu treatment for 12 h significantly increased the AsA content in kiwifruit leaves and up-regulated key genes involved in AsA biosynthesis, such as GDP-L-galactose phosphorylase3 (GGP3) and GDP-mannose-3′,5′-epimerase (GME). Overexpressing GGP3 in transgenic kiwifruit significantly increased the endogenous AsA content of kiwifruit, which was beneficial in mitigating Cu toxicity by decreasing levels of reactive oxygen species, malondialdehyde, and electrolyte leakage, as well as reducing damage to the chloroplast structure and photosystem II. This study presented a novel strategy to ameliorate plant Cu stress by increasing the endogenous antioxidant (AsA) content through transgenesis.