Copper Accumulation Research Articles

The Role of Glia in Wilson's Disease: Clinical, Neuroimaging, Neuropathological and Molecular Perspectives.

Wilson's disease (WD) is inherited in an autosomal recessive manner and is caused by pathogenic variants of the ATP7B gene, which are responsible for impaired copper transport in the cell, inhibition of copper binding to apoceruloplasmin, and biliary excretion. This leads to the accumulation of copper in the tissues. Copper accumulation in the CNS leads to the neurological and psychiatric symptoms of WD. Abnormalities of copper metabolism in WD are associated with impaired iron metabolism. Both of these elements are redox active and may contribute to neuropathology. It has long been assumed that among parenchymal cells, astrocytes have the greatest impact on copper and iron homeostasis in the brain. Capillary endothelial cells are separated from the neuropil by astrocyte terminal legs, putting astrocytes in an ideal position to regulate the transport of iron and copper to other brain cells and protect them if metals breach the blood-brain barrier. Astrocytes are responsible for, among other things, maintaining extracellular ion homeostasis, modulating synaptic transmission and plasticity, obtaining metabolites, and protecting the brain against oxidative stress and toxins. However, excess copper and/or iron causes an increase in the number of astrocytes and their morphological changes observed in neuropathological studies, as well as a loss of the copper/iron storage function leading to macromolecule peroxidation and neuronal loss through apoptosis, autophagy, or cuproptosis/ferroptosis. The molecular mechanisms explaining the possible role of glia in copper- and iron-induced neurodegeneration in WD are largely understood from studies of neuropathology in Parkinson's disease and Alzheimer's disease. Understanding the mechanisms of glial involvement in neuroprotection/neurotoxicity is important for explaining the pathomechanisms of neuronal death in WD and, in the future, perhaps for developing more effective diagnostic/treatment methods.

A strategy of “adding fuel to the flames” enables a self-accelerating cycle of ferroptosis-cuproptosis for potent antitumor therapy

Cuproptosis in antitumor therapy faces challenges from copper homeostasis efflux mechanisms and high glutathione (GSH) levels in tumor cells, hindering copper accumulation and treatment efficacy. Herein, we propose a strategy of “adding fuel to the flames” for potent antitumor therapy through a self-accelerating cycle of ferroptosis-cuproptosis. Disulfiram (DSF) loaded hollow mesoporous copper-iron sulfide (HMCIS) nanoparticle with conjugation of polyethylene glycol (PEG) and folic acid (FA) (i.e., DSF@HMCIS-PEG-FA) was developed to swiftly release DSF, H2S, Cu2+, and Fe2+ in the acidic tumor microenvironment (TME). The hydrogen peroxide (H2O2) levels and acidity within tumor cells enhanced by the released H2S induce acceleration of Fenton (Fe2+) and Fenton-like (Cu2+) reactions, enabling the powerful tumor ferroptosis efficacy. The released DSF acts as a role of “fuel”, intensifying catalytic effect (“flame”) in tumor cells through the sustainable Fenton chemistry (i.e., “add fuel to the flames”). Robust ferroptosis in tumor cells is characterized by serious mitochondrial damage and GSH depletion, leading to excess intracellular copper that triggers cuproptosis. Cuproptosis disrupts mitochondria, compromises iron-sulfur (Fe–S) proteins, and elevates intracellular oxidative stress by releasing free Fe3+. These interconnected processes form a self-accelerating cycle of ferroptosis-cuproptosis with potent antitumor capabilities, as validated in both cancer cells and tumor-bearing mice.

Glutathione-Scavenging Celastrol-Cu Nanoparticles Induce Self-Amplified Cuproptosis for Augmented Cancer Immunotherapy.

Cuproptosis is a novel copper-dependent programmed cell death. The efficacy of cuproptosis is highly dependent on intracellular copper accumulation and counteracted by a high level of glutathione (GSH) in tumor cells. Here, this work develops a self-amplified cuproptosis nanoparticles (Cel-Cu NP) using celastrol (Cel), a natural product isolated from medical plant. In Cel-Cu NP, Cel serves as a versatile copper ionophore, exhibiting an ideal coordination capacity toward copper ions without compromising the cuproptosis induction. Notably, Cel can simultaneously scavenge GSH content to amplify cuproptosis. Moreover, this self-amplified cuproptosis further activates immunogenic cell death (ICD) to elicit robust immune response. Combining with immune checkpoint blockade, Cel-Cu NP effectively eradicates metastatic tumors in a mouse lung metastasis model. This study provides an efficient nanomedicine by inducing self-amplified cuproptosis for robust immunotherapy.

Long-read sequencing identifies an SVA_D retrotransposon insertion deep within the intron of ATP7A as a novel cause of occipital horn syndrome

BackgroundSINE-VNTR-Alu (SVA) retrotransposons move from one genomic location to another in a ‘copy-and-paste’ manner. They continue to move actively and cause monogenic diseases through various mechanisms. Currently, disease-causing SVA retrotransposons...

Hepatic T‐cell lymphoma in a dog with chronic hepatitis and presence of Leptospira spp. in hepatic tissue

AbstractA 7‐year‐old, female, neutered Border collie was referred for investigation of increased liver enzymes, accompanied by mild non‐specific clinical signs. Investigations subsequently revealed the presence of chronic pyogranulomatous hepatitis, copper accumulation and presence of multiplying Leptospira spp. in the liver tissue. Following treatment with doxycycline and D‐penicillamine, the liver parameters and clinical signs improved. However, 9 months after the treatment was initiated, the dog re‐presented for overall deterioration, small intestinal diarrhoea and cough. Investigations were consistent with a hepatic large cell lymphoma. The dog was euthanased shortly after diagnosis, and histopathology of the postmortem samples taken from the liver showed presence of CD3+, CD11d+ T‐cell lymphoma as well as multiplying Leptospira spp. in the tissue.

Construction and validation of a prognostic and therapeutic cuproptosis- and immune-related gene signature in hepatocellular carcinoma.

Abnormal accumulation of copper could induce cell death and tumor growth, and affect tumor immune escape by regulating programmed cell death ligand 1 (PD-L1) expression. This study aims to establish and verify a risk signature based on cuproptosis- and immune-related genes (CIRGs) for hepatocellular carcinoma (HCC) management. HCC RNA-seq and clinical data were obtained from open databases. Least absolute shrinkage and selection operator (LASSO) and Cox regression analyses were utilized to screen CIRGs and develop a risk signature. The signature's value for clinical applications, functional enrichment, tumor mutation burden (TMB), and immune profile analyses were investigated systematically. A risk signature was developed utilizing seven CIRGs, and it performed well in predicting the prognosis of HCC patients in both the training and external validation cohorts. The model's risk score was discovered to be related to important clinical features. Top 15 mutated genes in HCC were significantly different among different risk groups. High-risk patients showed higher TMB, and high TMB was closely identified with a poorer prognosis. Immune profile analyses showed that immune infiltration level was higher in low-risk patients than high-risk patients, and the level of immune checkpoint genes expression varied significantly between patients in two different risk groups. Low-risk patients responded well to immunotherapy treatment, whereas high-risk patients were more sensitive to sorafenib, doxorubicin, gemcitabine and AKT (also known as protein kinase B) inhibitors. The established risk signature based on CIRGs can not only well predict the prognosis of HCC patients but is also promising in evaluating TMB and treatment response to immunotherapy, targeted therapy and chemotherapy, which has the potential to assist in the clinical management of HCC.

The Effects of Different Combinations of Cattle Organic Soil Amendments and Copper on Lettuce (cv. Rufus) Plant Growth

In modern agricultural production, cattle manure waste recovery is considered as a sustainable approach to agricultural waste management, reducing environmental pollution and chemical fertilizer use. This study aimed to investigate the effects of manure and digestate derived from a pilot-scale livestock waste-recycling system, in combination with a low copper concentration as a fungicide, on the physiological response of lettuce cv Rufus (Lactuca sativa L.) plants and the associated soil microbiome. A five-week microcosm experiment was conducted in a greenhouse under environmental conditions. Lettuce plant performance was assessed in terms of biomass, leaf area index, photosynthetic activity, chlorophyll measurements, lipid peroxidation, total phenolic content, and nutrient uptake. The results suggested that incorporating digestate into the potting soil mix significantly enhanced crop yields compared to the control and manure treatments. The soil microbial activity increased in the presence of fertilizers, improving the soil chemical and biological properties. The addition of copper negatively affected the growth and physiological performance of the lettuce plants under both the control and manure-treated conditions, except for those grown in the presence of digestate, where copper accumulation was reduced. These findings highlight the potential of growing horticultural crops using organic fertilization through livestock waste anaerobic digestate, establishing a waste-to-food recycling system.

Nighttime Warming Reduced Copper Concentration and Accumulation in Wheat Grown in Copper-Contaminated Soil by Affecting Physiological Traits

The changes in biomass (including yield), copper (Cu) concentration, and the accumulation of wheat (Triticum aestivum. L) in response to soil Cu pollution under nighttime warming had still not been explored. Hence, this study was carried out, and these variations were analyzed from a physiological perspective. Pot trials were performed at two levels of ambient temperatures (no-warming (NT) and average nighttime warming of 0.28 °C (WT)) and two levels of soil Cu concentrations (control check without Cu application (CK) and 100 mg/kg Cu application (Cu)). Soil was collected from the carbonate cinnamon soil region of central China. The warming effects of the passive nighttime warming system were prominent, and the average increment was 0.28 °C. Antioxidant enzyme activities were promoted by warming (p < 0.05) and Cu. The highest yield was achieved in NT-Cu, mainly attributed to relatively strong root activity and photosynthesis caused by supplemental Cu, but the Cu concentration in its grains was close to the threshold (10 mg/kg) for Cu concentration in foodstuff and could present a potential food safety risk. Though nighttime warming did not increase the total biomass and yield of wheat, it decreased the Cu accumulation of wheat grown in Cu-contaminated soil, especially in grains. Moreover, WT-CK and WT-Cu increased the Cu concentration in the roots and glumes and reduced the Cu concentration in grains by 13.09% and 55.84%, respectively, probably because of a lower transpiration rate. Among them, the Cu concentration of grains in WT-Cu was the lowest and significantly lower than other applications. Our findings reveal that nighttime warming has the potential to reduce the Cu risk of grains in wheat grown in the Cu-contaminated carbonate cinnamon soil region of central China and could then provide a theoretical reference for risk assessment of food quality for wheat subjected to dual stress from nighttime warming and Cu pollution.

Assessment of the Cu phytoremediation potential of Chrysanthemum indicum L. and Tagetes erecta L. using analysis of growth and physiological characteristics.

In the current study, the Cu phytoremediation ability of two ornamental plants, Chrysanthemum indicum L. and Tagetes erecta L., was tracked concerning the growth and physiological responses. Plants were subjected to varying concentrations of Cu (0, 100, 200, and 400 mg/kg) under the pot experiment for 8 weeks. The results showed that the measured growth and physiological characteristics declined in T. erecta shoots and roots at all tested treatments compared with the control. However, in C. indicum at 100 mg/kg, shoot biomass, shoot total soluble protein, and leaves number remained equal to that of the control and then reduced by rising Cu concentrations, compared with the control. Also, results indicated that in C. indicum, after 56 days of exposure to Cu, the chlorophyll pigments content markedly increased and reached a maximum level at 100 mg/kg dose and gradually declined with enhancing Cu concentrations, compared with the control. Other measured growth and physiological parameters decreased in both tissues of C. indicum in response to Cu usage in the growth medium. The carotenoid content of T. erecta decreased in all studied Cu levels in comparison to the control, but in C. indicum remained unaffected up to 200 mg/kg Cu in comparison to the control and then enhanced with increasing Cu level. The augmentation of antioxidant enzyme activity in two species, especially in roots, reflected the incident of Cu stress as demonstrated by elevated MDA and ion leakage levels. Data concerning copper accumulation in tissues, TF, and BAF showed T. erecta is a weak Cu accumulator and seems not to be an appropriate candidate for Cu phytoremediation. However, the Cu content in shoots and roots of C. indicum increased significantly with an increment in applied Cu level. Also, C. indicum accumulated higher Cu concentrations in the roots than in shoots and exhibited TF < 1, 0.1 < BAF root < 1, and can be considered as a Cu excluder by the phytostabilization mechanism.

A Case Report on Wilson’s Disease

The accumulation of copper in the liver, brain, cornea, and kidneys is the hallmark of Wilson's disease, an uncommon autosomal recessive condition. There is no community-based research on the prevalence and incidence of Wilson's disease in India; this study is hospital-based. Overview of the Case: A 10-year-old girl with serious complaints of burning micturition, giddiness, loose stools and blood in stools, yellowish discolouration of skin and sclera, generalized body aches, fever and stomach pain with distension was brought to the paediatric department. Wilson's illness was established by abdominal USG using the increased levels of urine copper, whole blood picture, liver function tests, and serum electrolytes and decreased levels of ceruloplasmin. The prominent characteristic, which is less common in youngsters, is the Kayser Fleisher ring. It is identified by a discoloration of the corneal edge that is greenish-brown in colour and eventually goes away with treatment. Probiotics, an antibiotic, a hepatoprotective drug, and copper chelators (D-penicillamine and zinc) were used in her treatment. Gradually she showed improvement in clinical signs and LFT levels.

The gene IFIT1 is associated with dietary copper‐induced yellow fat disease in sheep

AbstractIn the process of rapid fattening and rearing of meat sheep, yellow fat disease of sheep occurs frequently. This study aims to investigate the preliminary pathogenesis of yellow fat disease in sheep. Eighteen healthy sheep (4–5 months old, 34 ± 1 kg) were selected and randomly divided into three groups: the 10 ppm copper group, the 50 ppm copper group, and the 100 ppm copper group. At the end of the experiment, blood, liver, kidney, and adipose tissue samples were taken from all sheep, and measurements of each index were taken. 50 and 100 ppm copper supplementation in the diets did not significantly affect average daily gain, total cholesterol (TC), triglyceride (TG) and sorbitol dehydrogenase in sheep but significantly increased the effects on gamma‐glutamyltransferase, aspartate aminotransferase, and alanine aminotransferase enzyme activities in the liver and increased the accumulation of copper in the liver. 50 and 100 ppm copper supplementation to the feed caused different levels of pathological damage to the liver, the kidney, and fat and significantly affected the brightness, redness, and yellowness of the carcass fat. Sheep in the 50 ppm copper group did not show significant clinical symptoms of yellow fat disease in the later period of the experiment, but those in the 100 ppm copper group showed significant clinical symptoms of yellow fat disease. Transcriptome analysis of sheep livers showed differential genes associated with yellow fat disease, and GO and KEGG analyses associated with yellow fat disease were performed, and further correlation analysis found that the occurrence of copper‐induced yellow fat disease may be closely related to gene IFIT1.

A Stimulus-Responsive Ternary Heterojunction Boosting Oxidative Stress, Cuproptosis for Melanoma Therapy.

Cuproptosis, a recently discovered copper-dependent cell death, presents significant potential for the development of copper-based nanoparticles to induce cuproptosis in cancer therapy. Herein, a unique ternary heterojunction, denoted as HACT, composed of core-shell Au@Cu2O nanocubes with surface-deposited Titanium Dioxide quantum dots and modified with hyaluronic acid is introduced. Compared to core-shell AC NCs, the TiO2/Au@Cu2O exhibits improved energy structure optimization, successfully separating electron-hole pairs for redox use. This optimization results in a more rapid generation of singlet oxygen and hydroxyl radicals triggering oxidative stress under ultrasound radiation. Furthermore, the HACT NCs initiate cuproptosis by Fenton-like reaction and acidic environment, leading to the sequential release of cupric and cuprous ions. This accumulation of copper induces the aggregation of lipoylated proteins and reduces iron-sulfur proteins, ultimately initiating cuproptosis. More importantly, HACT NCs show a tendency to selectively target cancer cells, thereby granting them a degree of biosecurity. This report introduces a ternary heterojunction capable of triggering both cuproptosis and oxidative stress-related combination therapy in a stimulus-responsive manner. It can energize efforts to develop effective melanoma treatment strategies using Cu-based nanoparticles through rational design.

Copper Accumulation and its Effect on Glyoxalase System, AsA-GSH Cycle, Antioxidase Activity in Pontederia cordota Leaves

Copper Accumulation and its Effect on Glyoxalase System, AsA-GSH Cycle, Antioxidase Activity in Pontederia cordota Leaves

Treatment of marble industry wastewater by Brassica napus (L.) under oxalic acid amendment: efficacy as fodder and carcinogenic risk assessment.

In the present study, Brassica napus, a food plant, was grown for phytoextraction of selected heavy metals (HMs) from marble industry wastewater (WW) under oxalic acid (OA) amendment. The hydroponic experiment was performed under different combination of WW with OA in complete randomized design. Photosynthetic pigments and growth reduction were observed in plants treated with WW alone amendments. The combination of OA in combination with WW significantly enhanced the growth of plants along with antioxidant enzyme activities compared with WW-treated-only plants. HM stress alone enhanced the hydrogen peroxide, electrolyte leakage, and malondialdehyde contents in plants. OA-treated plants were observed with enhanced accumulation of cadmium (Cd), copper (Cu), and lead (Pb) concentrations in the roots and shoots of B. napus. The maximum concentration and accumulation of Cd in root, stem, and leaves was increased by 25%, 30%, and 30%; Cu by 42%, 24%, and 17%; and Pb by 45%, 24%, and 43%, respectively, under OA amendment. Average daily intake and hazard quotient (HQ) were calculated for males, females, and children in two phases of treatments in phytoremediation of metals before and after accumulation into B. napus leaves and stems. HQ of metals in the leaves and stem was < 1 before metal accumulation, whereas > 1 was observed after HM accumulation for all males, females, and children. Similarly, the hazard index of the three study types was found > 1. It was observed that the estimated excess lifetime cancer risk was of grade VII (very high risk), not within the accepted range of 1 × 10-4 to 1 × 10-6. Based on the present study, the increased levels of HMs up to carcinogenicity was observed in the B. napus which is not safe to be consumed later as food.

Sleep fragmentation exacerbates myocardial ischemia‒reperfusion injury by promoting copper overload in cardiomyocytes

Sleep disorders increase the risk and mortality of heart disease, but the brain-heart interaction has not yet been fully elucidated. Cuproptosis is a copper-dependent type of cell death activated by the excessive accumulation of intracellular copper. Here, we showed that 16 weeks of sleep fragmentation (SF) resulted in elevated copper levels in the male mouse heart and exacerbated myocardial ischemia–reperfusion injury with increased myocardial cuproptosis and apoptosis. Mechanistically, we found that SF promotes sympathetic overactivity, increases the germination of myocardial sympathetic nerve terminals, and increases the level of norepinephrine in cardiac tissue, thereby inhibits VPS35 expression and leads to impaired ATP7A related copper transport and copper overload in cardiomyocytes. Copper overload further leads to exacerbated cuproptosis and apoptosis, and these effects can be rescued by excision of the sympathetic nerve or administration of copper chelating agent. Our study elucidates one of the molecular mechanisms by which sleep disorders aggravate myocardial injury and suggests possible targets for intervention.

Non-ceruloplasmin copper and urinary copper in clinically stable Wilson disease: Alignment with recommended targets

Background and AimsWilson disease (WD) is caused by accumulation of copper primarily in liver and brain. During maintenance therapy of WD with D-penicillamine, current guidelines recommend on-treatment ranges of urinary copper excretion (UCE) 200-500 μg/24h and serum non-ceruloplasmin bound copper (NCC) 50-150 μg/L. We compared NCC measured by two novel assays and UCE from clinically stable WD patients on D-penicillamine therapy with these recommendations. MethodsThis is a secondary analysis of data from the Chelate Trial (NCT03539952) that enrolled physician selected clinically stable WD patients on D-penicillamine maintenance therapy with unaltered dose for at least four months. We analyzed laboratory samples from the first screening visit, prior to interventions. NCC was measured by either protein speciation (NCC-Sp) using anion exchange HPLC protein speciation followed by copper determination with ICP-MS or as exchangeable copper (NCC-Ex). NCC-Sp was also analyzed in healthy controls (n=75). ResultsIn 76 WD patients with 21.3±14.3 average treatment-years, NCC-Sp (Mean±SD: 56.6±26.2 μg/L) and NCC-Ex (Mean±SD: 57.9±24.7 μg/L) were within the 50-150 μg/L target in 61% and 54%, respectively, below in the remaining. In addition, 36% and 31%, respectively, were even below the normal ranges (NCC-Sp: 46-213 μg/L, NCC-Ex: 41-71 μg/L). NCC-Ex positively correlated with NCC-Sp (r2=0.66, P<0.001) but with systematic deviation. UCE was outside the 200-500 μg/24H target in 58%. Only 14/69 (20%) fulfilled both the NCC-Sp and UCE targets. Clinical or biochemical signs of copper deficiency were not detected. ConclusionClinically stable WD patients on maintenance D-penicillamine therapy frequently have lower NCC-Sp or higher UCE than current recommendations without signs of overtreatment. Further studies are warranted to identify appropriate target ranges of NCC-Sp, NCC-Ex and UCE in treated WD. Impact and implicationsChelator treatment of patients with Wilson Disease (WD) is currently guided by measurements of non-ceruloplasmin bound copper (NCC) and 24 h urinary copper excretion (UCE) but validation is limited. In 76 adults with ≈21 years history of treated WD and clinically stable disease on D-penicillamine therapy, NCC was commonly found to be below normal values and recommended target ranges whether measured by protein speciation (NCC-Sp) or as exchangeable copper (NCC-Ex) while UCE values were above recommended target range in 49%. Common wisdom would suggest overtreatment in these cases but no clinical or biochemical signs of copper deficiency were observed. Exploratory analysis of liver enzymes suggested that NCC below levels seen in controls may be beneficial, while the relation to UCE was less clear. The data calls for critical re-evaluation of target ranges for treatment of WD, specific for drug and laboratory methodology. Clinical trial number(NCT03539952)

Does transplanted Posidonia oceanica act as a sink or source of trace elements? Ecological implications for restoring polluted coastal areas

Despite the high potential of seagrass restoration to reverse the trend of marine ecosystem degradation, there are still many limitations, especially when ecosystems are severely degraded. In particular, it is not known whether restoring polluted ecosystems can lead to potentially harmful effects associated with contaminant remobilisation. Here, we aimed to investigate the role of P. oceanica transplanted from a pristine meadow to a polluted site (Augusta Bay, Italy, Mediterranean Sea) in two seasons of the year, as a sink or source of trace elements to the environment. The main results showed i) higher accumulation of chromium (Cr), copper (Cu) and total mercury (THg) in plants transplanted in summer than in winter, as well as an increase in Cr and THg in plants from sites with higher trace element loads; ii) an increase in leaf phenolics and a decrease in rhizome soluble carbohydrates associated with As and THg accumulation, suggesting the occurrence of defence strategies to cope with pollution stress; iii) a different partitioning of trace elements between below- and above-ground tissues, with arsenic (As) and Cr accumulating in roots, whereas Cu and THg in both roots and leaves. These results suggest that P. oceanica transplanted to polluted sites can act as both a sink and a source, sequestering trace elements in the below-ground tissues thus reducing their bioavailability, but also potentially remobilising them. However, the amount of trace elements potentially exported from P. oceanica to the environment through transfer into food webs via leaves and detritus appeared to be low under the specific conditions of the study site. Although further research into seagrass restoration of polluted sites would improve current knowledge to support effective ecosystem-based coastal management, the benefits of restoring polluted sites through seagrass transplantation appear to outweigh the potential costs of inaction over time.

Copper hepatopathies in Australian dogs.

To evaluate hepatopathies in Australian dogs according to the World Small Animal Veterinary Association (WSAVA) guidelines. Specifically, to describe the prevalence and survival of dogs with copper-associated hepatopathy. Medical records from the Small Animal Specialist Hospital were reviewed to identify dogs with liver disease and liver biopsy between November 2008 and November 2021. Liver histopathology reports were reviewed with a board-certified veterinary pathologist and classified according to the WSAVA guidelines. Histopathology reports and clinical records were reviewed to ascertain the most important histological process for statistical analysis. Copper-associated hepatopathy was defined as (i) histological evidence of copper accumulation in centrilobular areas (Zone 3) associated with hepatocyte necrosis, inflammation with copper-laden macrophages and chronic hepatitis (ii) histochemical copper staining showing hepatocyte copper accumulation in the centrilobular areas and iii) hepatic copper measurement with concentrations greater than 600 μg/g dry weight of liver. Dogs with primary inflammatory parenchymal disease included dogs with copper-associated hepatopathy, idiopathic chronic hepatitis, non-specific reactive hepatitis, chronic bacterial hepatitis and immune-mediated chronic hepatitis. Descriptive statistics were performed for all dogs. Age, weight and clinicopathologic data were compared between dogs with copper-associated hepatopathy and dogs with other causes of chronic primary inflammatory parenchymal liver disease (Kruskal-Wallis test). Survival times were calculated and compared (Kaplan-Meier curves and log rank test) between dogs with copper-associated hepatopathy and dogs with other chronic primary inflammatory parenchymal liver diseases. Breed was evaluated to determine the breed most commonly affected with copper-associated hepatopathy and identify any breed in which this disease has not previously been described. Sixty-seven (43 female, 24 male) dogs with a median age of 7.8 years (quartile [Q] Q1-Q3 4.5-9.6 years) were included. Thirteen dogs had copper-associated hepatopathy, eight dogs had idiopathic chronic hepatitis, eight dogs had non-specific reactive hepatitis, seven dogs had disorders associated with portal hypertension, five dogs had chronic bacterial hepatitis and four dogs had immune-mediated chronic hepatitis. Compared with dogs with other causes of chronic primary inflammatory parenchymal liver disease, dogs with copper-associated hepatopathy tended to be younger (6.73 vs. 8.01 years, P = 0.057) and heavier (19.8 vs. 9.6 kg, P = 0.052) than dogs with other causes of primary chronic inflammatory parenchymal diseases. There was no statistically significant difference when ALT (P = 0.30), ALP (P = 0.18) and total bilirubin (P = 0.13) were compared between the two groups. The median survival time for all dogs after liver biopsy was 2010 days (CI 1321 days - not reached). There was no significant difference in survival between dogs with copper-associated hepatopathy and dogs with other causes of chronic primary inflammatory parenchymal liver disease (P = 0.5). Copper-associated hepatopathy was common among Australian dogs with chronic hepatopathies, occurring in younger and heavier dogs than other causes of primary inflammatory parenchymal liver disease. Clinical pathology is not useful for differentiating between copper-associated hepatopathy and other causes of chronic primary inflammatory parenchymal liver disease. When copper-associated hepatopathy is treated, the prognosis can be good. This is the first report of copper-associated hepatopathy in Australian Cavalier King Charles Spaniels.

Wilson Disease: Copper-Mediated Cuproptosis, Iron-Related Ferroptosis, and Clinical Highlights, with Comprehensive and Critical Analysis Update.

Wilson disease is a genetic disorder of the liver characterized by excess accumulation of copper, which is found ubiquitously on earth and normally enters the human body in small amounts via the food chain. Many interesting disease details were published on the mechanistic steps, such as the generation of reactive oxygen species (ROS) and cuproptosis causing a copper dependent cell death. In the liver of patients with Wilson disease, also, increased iron deposits were found that may lead to iron-related ferroptosis responsible for phospholipid peroxidation within membranes of subcellular organelles. All topics are covered in this review article, in addition to the diagnostic and therapeutic issues of Wilson disease. Excess Cu2+ primarily leads to the generation of reactive oxygen species (ROS), as evidenced by early experimental studies exemplified with the detection of hydroxyl radical formation using the electron spin resonance (ESR) spin-trapping method. The generation of ROS products follows the principles of the Haber-Weiss reaction and the subsequent Fenton reaction leading to copper-related cuproptosis, and is thereby closely connected with ROS. Copper accumulation in the liver is due to impaired biliary excretion of copper caused by the inheritable malfunctioning or missing ATP7B protein. As a result, disturbed cellular homeostasis of copper prevails within the liver. Released from the liver cells due to limited storage capacity, the toxic copper enters the circulation and arrives at other organs, causing local accumulation and cell injury. This explains why copper injures not only the liver, but also the brain, kidneys, eyes, heart, muscles, and bones, explaining the multifaceted clinical features of Wilson disease. Among these are depression, psychosis, dysarthria, ataxia, writing problems, dysphagia, renal tubular dysfunction, Kayser-Fleischer corneal rings, cardiomyopathy, cardiac arrhythmias, rhabdomyolysis, osteoporosis, osteomalacia, arthritis, and arthralgia. In addition, Coombs-negative hemolytic anemia is a key feature of Wilson disease with undetectable serum haptoglobin. The modified Leipzig Scoring System helps diagnose Wilson disease. Patients with Wilson disease are well-treated first-line with copper chelators like D-penicillamine that facilitate the removal of circulating copper bound to albumin and increase in urinary copper excretion. Early chelation therapy improves prognosis. Liver transplantation is an option viewed as ultima ratio in end-stage liver disease with untreatable complications or acute liver failure. Liver transplantation finally may thus be a life-saving approach and curative treatment of the disease by replacing the hepatic gene mutation. In conclusion, Wilson disease is a multifaceted genetic disease representing a molecular and clinical challenge.

Comparative efficacy of stannous fluoride and zinc sulfate solution to copper sulfate footbath solutions for the treatment and prevention of digital dermatitis in lactating dairy cows

The objective of this study was to determine whether a novel footbath solution containing stannous fluoride (SnF2) was superior to 5% copper sulfate solution for the treatment and prevention of digital dermatitis (DD) in dairy cattle. Study 1 was conducted over 4 weeks in Missouri and involved 34 lactating Holstein-Friesian cows with hind feet DD lesions. Cows in group SF walked through a footbath containing a proprietary formulation of SnF2 once weekly, whereas cows in group CS walked through a 5% CuSO4 footbath once daily for 5 d each week. Study 2 was conducted over 8 weeks in California and involved 40 lactating Holstein-Friesian cows with hind feet DD lesions. Cows in group SF walked through a SnF2 footbath for 3 consecutive days then once a week for the following 7 weeks. Cows in group CS walked through a 5% CuSO4 footbath 3 times each week for 8 weeks. Data collection included lesion type, lesion area, locomotion score, and pain score. Digital dermatitis was actively transmitted in study 1, and lesion area and locomotion scores were lower in group SF than group CS. In contrast, DD was not actively transmitted in study 2, and lesion area and locomotion scores were similar in groups SF and CS. Stannous fluoride delayed the development of active DD lesions in study 1 compared with copper sulfate, with a lower relative risk (0.57, P < 0.001) of a hind foot developing an active DD lesion over 28 d. However, SnF2 decreased the rate that active DD lesions transitioned to M3, M4, or M0 lesions compared with 5% copper sulfate in both studies, with the relative risk of a hind foot with an active DD lesion transitioning to M3, M4, or M0 in group SF being slightly lower in study 1 (0.83, P = 0.042) and study 2 (0.90, P = 0.020) than group CS. Our findings demonstrated that walking cows through a stannous fluoride footbath once per week in a herd undergoing active transmission of infection was more effective in preventing active DD lesions, but less effective in treating active DD lesions, than walking cows through a copper sulfate footbath 4 times per week. The novel SnF2 footbath solution shows promise for controlling DD in dairy herds that want an alternative footbath solution to CuSO4 and are interested in limiting the environmental accumulation of copper.