Liver Copper Levels Research Articles

Unprecedented Co-occurrence: Identification of a Pathogenic Genetic Variant in the KMT2B Gene in a Wilson Disease Patient with a Pathogenic ATP7B Mutation

The pathophysiology of dystonia in Wilson disease (WD) is complex and poorly understood. Copper accumulation in the basal ganglia, disrupts dopaminergic pathways, contributing to dystonia’s development via neurotransmitter imbalance. Despite advances in diagnosis and management, WD with dystonia remains a challenging condition to treat. We aim to report the unprecedented co-occurrence of pathogenic genetic variants in both the ATP7B and KMT2B genes in a patient with WD. A 13-year-old male presented at 12 with dysarthria and bilateral Kayser-Fleischer rings. Over months, dystonia spread to his left foot, upper limb, and trunk, accompanied by slowed daily activities. Diagnostic tests included MRI for brain structure, abdominal ultrasound for liver function, serum ceruloplasmin and copper levels to assess copper metabolism, and 24-hour urine copper tests for excretion levels. Whole exome sequencing was conducted using genomic DNA from peripheral blood samples. Variant classification followed guidelines from the American College of Medical Genetics and Genomics. The sequencing revealed compound heterozygous pathogenic variants in the ATP7B gene: NM_000053.4:c.2165dupT and NM_000053.4:c.813C>A. A pathogenic variant in the KMT2B gene, NM_014727:c.3052delA, was identified. This case highlights WD co-occurrence with ATP7B and KMT2B mutations, suggesting KMT2B as a potential genetic modifier.

Oral iron supplementation ameliorated alterations in iron uptake and utilization in copper-toxic female Wistar rats.

Women are more susceptible to both iron deficiency and copper toxicity due to monthly flow and estrogen action, respectively. Oral iron is beneficial for menstruating women and enhances erythropoiesis, but both deficiency and excess of copper impact iron absorption and mobilization. The aim of this study was to investigate the possibility of mitigating copper toxicity in female Wistar rats while supplementing with iron. 20 female rats (160-180g) were grouped into four: Groups 1 (Control) received 0.3mls normal saline, 2- copper-toxic (100m mg/kg Copper sulphate), 3- Copper-toxic+Iron (100 mg/kg Copper sulphate + 1 mg/kg Ferrous sulphate) and 4- Iron (1 mg/kg Ferrous sulphate). All treatment was administered orally for 5 weeks. Blood was collected retro-orbitally after light anesthesia into EDTA and plain bottles for hematological, serum copper, iron, ferritin and total iron binding capacity (TIBC) analysis. Liver was excised for copper and iron levels while bone marrow was harvested for myeloid/erythroid ratio. The data were analyzed by one-Way ANOVA and statistical significance was considered at p<0.05. Iron supplementation significantly increased packed cell volume, hemoglobin concentration, red blood cell count and myeloid/erythroid ratio, compared to the copper-toxic group. Serum iron and TIBC were significantly increased while liver copper and iron levels reduced significantly in iron supplemented group compared to the copper-toxic group. Oral iron supplementation mitigated alterations in iron absorption and mobilization following copper toxicity.

Environmental impacts of brass mesh nets on open ocean aquaculture pens in tropical marine environments

Aquaculture pens with brass mesh were installed at commercial fish farms in two locations, Panama and Mexico. Measurements of copper and zinc levels in the water and sediments were monitored over 468 and 435 days for the Panama and Mexico sites respectively. Additional chemical parameters were also measured including dissolved organic carbon in the water column and total solids, total organic carbon, total sulfides, redox potential and acid volatile sulfides and simultaneously extracted metals in the sediments. Measurements were taken at a reference site 1000 m away from the pens perpendicular to the dominant current direction, as well as 50 m upstream, 50 m downstream and adjacent to the pens. Muscle, skin and liver samples were taken from cobia (Rachycentron canadum) produced at the Panama site and analyzed for copper content.The dissolved copper levels at both farms stayed below 1 μg/L at all times. There was no significant difference in the dissolved copper levels between the reference site and other measurement sites at either location. There was also no observable increase in dissolved copper levels over time at the Mexico location although there was a significant difference in dissolved copper levels between the sampling days at the Panama location. Zinc concentrations also remained within safe levels, never exceeding 0.82 μg/L at any point. Copper and zinc levels in the sediments did not show a significant increase over time. The copper levels in tissue samples from the muscle tissue of the cobia raised in a brass mesh pen at the Panama site were 19% higher than those raised in the plastic mesh pen. The elevated copper levels are well below recommended daily intake levels and do not pose a risk to human health. There was no significant difference in copper levels of the skin or liver tissue samples. The absence of any increase in liver copper levels suggests that the fish's physiology and welfare are not impacted by brass mesh.The findings suggest that brass mesh is environmentally compatible with high energy marine environments and a commercially viable product for open ocean aquaculture farms.

2366 Acute Liver Failure Caused by Hemophagocytic Lymphohistiocytosis Secondary to Aggressive Natural Killer Cell Leukemia and Epstein-Barr Virus Reactivation

INTRODUCTION: Acute liver injury is a common presentation of many systemic illnesses, including hematologic malignancies. The present case was an unusual presentation of acute liver failure (ALF) caused by hemophagocytic lymphohistiocytosis (HLH) secondary to aggressive natural killer cell leukemia(ANKL) associated with Epstein-Barr virus(EBV) infection reactivation. CASE DESCRIPTION/METHODS: A 24-year-old female presented with fever and altered mental status. Her physical examination revealed massive hepatosplenomegaly. Complete blood count showed pancytopenia with WBC 460/µL, microcytic anemia Hgb 7.8g/dL with MCV 79, thrombocytes 45,000/µL, AST 170IU/L, ALT 162IU/L, INR 1.5, albumin 1.3g/dl and total bilirubin 11.08mg/dL, triglycerides 1164mg/dL, ferritin 1792ng/ml. CT abdomen showed hepatosplenomegaly with hepatic steatosis. Her liver biopsy showed hepatic necrosis with moderate steatohepatitis and perisinusoidal fibrosis with normal liver copper level. She was initially treated with steroids and N-acetylcysteine. Flow cytometry and bone marrow biopsy revealed CD56+ and CD3+ positive cells with high serum soluble IL-2 receptor consistent with ANKL and HLH. EBV markers were found positive for EBV IgG antibody and EBV nuclear antigen with high EBV PCR and negative for EBV IgM antibody, which is consistent with EBV reactivation. A modified SMILE chemotherapy regimen with rituximab was initiated. Unfortunately, her clinical status continued to decline progressively with worsening acute encephalopathy, acute hypoxic respiratory failure and Pseudomonas septic shock which required vasopressors and IV antibiotics. The patient died on day-30 of hospitalization due to her hematologic malignancy, septic shock, and ALF. DISCUSSION: HLH is a consequence of abnormal immune activation which could be secondary to infections, malignancies, or autoimmune diseases; leading to life-threatening multi-organ inflammatory response, especially in the liver. Natural killer (NK) cells are types of lymphocytes that play an important role as part of innate immune system in fighting tumors and viral cells; NK cells are abundant in the liver. It was shown that NK cells might be involved in the intense immune response causing severe liver damage. ANKL is a rare malignant lymphoproliferative disorder of mature NK cells closely associated with EBV. EBV reactivation can occur in this disease setting. Careful evaluation of patients with ALF and abnormal hematologic profile is essential for early detection and therapy of the primary culprit.

A Gene Therapy Approach to Improve Copper Metabolism and Prevent Liver Damage in a Mouse Model of Wilson Disease.

Wilson disease (WD), an autosomal recessive disease caused by mutations in a copper-transporting P-type ATPase (Atp7b), causes severe liver damage. This disease is currently treated with the lifelong use of copper chelation therapy, which has side effects and does not fix copper metabolism. Here, we thoroughly characterized a mouse model of WD, the toxic milk mouse, and used the model to test a gene therapy approach for treating WD. WD mice accumulated copper in the liver from birth; severe copper accumulation and concurrent liver disease were evident by 2 months of age. Intravenously administering an adeno-associated viral (AAV) 8 vector expressing a codon-optimized version of the human ATP7B transgene into 2-month-old WD mice significantly decreased liver copper levels compared with age-matched, uninjected, WD mice. We also observed a significant dose-dependent decrease in liver disease. Male mice injected with 1011 genome copies of AAV8 vector showed only mild histopathological findings with a complete lack of liver fibrosis. Therefore, we conclude that administering gene therapy at the early stages of disease onset is a promising approach for reducing liver damage and correcting copper metabolism in WD.

Hepatotoxic and renal effects of the water soluble fractions of spent engine oil in Swiss albino mice

Spent engine oil (SEO) contains toxic metals which may be leached into water supplies during precipitation. These metals may bioaccumulate during exposure, eliciting adverse effects on the liver and the kidneys, both target organs for several contaminants. This study determined the hepatotoxic and renal effects of the water soluble fractions (WSF) of SEO in mice. The levels of renal function biomarkers (creatinine and urea) and iron, zinc, copper, nickel, lead and cadmium levels in livers of twenty-four male albino mice administered daily oral gavages of 0%, 1%, 10% and 100% WSF of SEO for 30 days were evaluated. Creatinine (0.17±0.06 mg/dl) and urea (22.33±1.53 mg/dl) levels were lowest and highest respectively at 10%, although values were marginal but were not significantly different from control (p>0.05). Iron (61150.27±2300.25ug/l), Zn (25942.57±975.86), Cu (7412.20±278.81), and Ni (185.33±6.98ug/l) levels in the liver were significantly elevated (p 0.05) compared with control and other treatments. Iron, zinc, copper and nickel levels at 10% were highest in the treatments. These results provide evidence for the adverse renal and hepatotoxic effects of water contaminated with SEO and highlight the need for its proper management. Keywords : Hepatotoxicity, creatinine, urea, metal contaminants, waste oil.

Changes in Liver Tissue Trace Element Concentrations During Hepatitis B Viral Infection Treatment.

Approximately 350-400 million people in the world have Hbs Ag (hepatitis B virus surface antigen) positivity. In the international guidelines, the permanent suppression of replication in chronic hepatitis B virus (HBV) infection therapy is reported as the primary therapeutic goal. Trace elements play a key role in liver diseases. The aim of our study is to determine some trace element concentrations in the liver during HBV treatment periods. The measurement of 11 trace elements (manganese, lead, nickel, chromium, cadmium, iron, copper, zinc, silver, cobalt, and aluminum) was carried out by the method of inductively coupled plasma mass spectrometry in liver biopsy materials (before starting treatment and at the sixth month of the treatment period). There was an increase in zinc and copper concentrations in liver materials at the sixth month of treatment compared to the pre-treatment values (the median zinc value was 48.05μg/g before treatment and 74.9μg/g at 6months after initial treatment, p = 0.035; median copper was 2.82μg/g before treatment and 5.31μg/g after 6months, p = 0.002). General estimations indicated that zinc (p = 0.002), iron (p = 0.0244), copper (p = 0.0003), and aluminum (p = 0.0239) values may be effective in HAI (histological activity index) changes. Only iron levels could be at a very low level effective on the changes caused by fibrosis (p = 0.0002). Liver tissue zinc and copper levels increased in parallel with the improvement of inflammation in antiviral-treated HBV patients. In addition, the levels of zinc and copper in the liver tissue can be useful markers for liver tissue damage detection.

High spatial resolution LA-ICP-MS demonstrates massive liver copper depletion in Wilson disease rats upon Methanobactin treatment

Wilson disease (WD) is a rare genetic disorder of the copper metabolism leading to systemic copper accumulation, predominantly in the liver. The therapeutic approach in WD patients is the generation of a negative copper balance and the maintenance of copper homeostasis, currently by the use of copper chelators such as D-penicillamine (D-PA). However, in circumstances of delayed diagnosis, poor treatment compliance, or treatment failure, mortality is almost certain without hepatic transplantation. Moreover, even after years of D-PA treatment, high liver copper levels are present in WD patients.We have recently suggested the use of the bacterial peptide Methanobactin (MB), which has an outstanding binding affinity for copper, as potentially efficient and patient-friendly remedy against copper damage in WD. Here we substantiate these findings considerably, by demonstrating a significant removal of copper from liver samples of WD rats upon short, one week only, MB treatments. Using laser ablation-inductively coupled plasma-mass spectrometry with a spatial resolution down to 4 μm, we demonstrate that only small copper hotspots remain in MB treated animal livers. We further demonstrate in WD rat liver, seven weeks after the stopped MB treatment, a lower liver copper concentration as compared to untreated control animals. Thus, MB highly efficiently depletes liver copper overload with a sustained therapeutic effect.

Complications of Liver Transplant in Adult Patients With the Hepatic Form of Wilson Disease.

Wilson disease is an autosomal, recessive, inherited disorder of copper metabolism that results in the accumulation of copper in many organs and tissues. This disease is mainly characterized by dysfunction due to copper accumulation in the liver, kidney, brain, cornea, bone, heart, and blood cells. The clinical spectrum is broad in Wilson disease. Asymptomatic Wilson disease may be present, but findings related to the involvement of an individual organ or multiple organ failure can be seen. These findings can include neurologic and neuropsychiatric complications. Our aim here was to examine the neurologic complications and our clinical experience in patients who underwent liver transplant for Wilson disease in our clinic. We retrospectively reviewed the medical records of transplant patients with Wilson disease who were seen at Baskent University Faculty of Medicine Transplantation Science between 2005 and 2017. Patient demographics, neurologic complaints, findings from neurologic examinations, and imaging findings were recorded. We also recorded the presence of the Kayser-Fleischer ring, serum ceruloplasmin, 24-hour copper urine levels, and levels of dry copper in liver in each patient. Our study included 19 patients who ranged in age range from 18 to 44 years (mean age of 26 years). Seven of 19 patients (36.8%) had neurologic symptoms, including epileptic seizures in 2 patients (10.5%), encephalopathy in 1 patient (5.2%), tremor in 3 patients (15.7%), and headache in 1 patient (5.2%). The cause of these long-term neurologic complications was the immunosuppressive drugs. Patients with epileptic seizures were provided with seizure control medication (levetiracetam). Tremor did not need treatment. In Wilson disease, neurologic complications can be severe. The most common complication seen in our patients was tremor. Early diagnosis and treatment may slow down neurologic disability.

Rat liver mitochondrial dysfunction by addition of copper(II) or iron(III) ions

Increased copper (Cu) and iron (Fe) levels in liver and brain are associated to oxidative stress and damage with increased phospholipid oxidation process. The aim of this work was to assess the toxic effects of Cu2+ and Fe3+ addition to rat liver mitochondria by determining mitochondrial respiration in states 3 (active respiration) and 4 (resting respiration), and phospholipid peroxidation. Both, Cu2+ and Fe3+ produced decreases in O2 consumption in a concentration-dependent manner in active state 3: both ions by 42% with malate-glutamate as complex I substrate (concentration for half maximal response (C50) 60μM Cu2+ and 1.25mM Fe3+), and with succinate as complex II substrate: 64–69% with C50 of 50μM Cu2+ and with C50 of 1.25mM of Fe3+. Respiratory control decreased with Cu2+ (C50 50μM) and Fe3+ (C50 1.25-1-75mM) with both substrates. Cu2+ produced a 2-fold increase and Fe3+ a 5-fold increase of thiobarbituric acid-reactive substances (TBARS) content from 25μM Cu2+ (C50 40μM) and from 100μM Fe3+ (C50 1.75mM). Supplementations with Cu2+ and Fe3+ ions induce mitochondrial dysfunction with phospholipid peroxidation in rat liver mitochondria. Although is proved that a Fenton/Haber Weiss mechanism of oxidative damage occurs in metal-ion induced mitochondrial toxicity, slightly different responses to the metal ions suggest some differences in the mechanism of intracellular toxicity. The decreased rates of mitochondrial respiration and the alteration of mitochondrial function by phospholipid and protein oxidations lead to mitochondrial dysfunction, cellular dyshomeostasis and cell death.

Accumulative Copper Poisoning in Sheep in Northeastern Brazil

Background: Despite the proven copper deficiency in several parts of Brazil, reports of acute and accumulative copper poisoning are increasing in ruminants. Sheep are known to be far more sensible to copper toxicity than cattle, but the sensibility of goats is not quite known. The present paper aimed to describe the epidemiological, clinical, laboratorial and pathological findings of two outbreaks of accumulative copper poisoning in sheep fed poultry litter in Ceará and Rio Grande do Norte, Northeastern Brazil, and to compare these findings with national outbreaks.Cases: Two outbreaks of copper poisoning occurred in the municipalities of Mossoró, Rio Grande do Norte [outbreak 1], and Icapuí, Ceará [outbreak 2], Brazil. A total of 35 sheep [outbreak 1] and 190 small ruminants (175 sheep and 15 goats) [outbreak 2] were fed poultry litter. First clinical signs of poisoning started after two months of litter consumption. Main clinical signs were apathy, jaundice, hepatomegaly, abdominal pain, diarrhea, and dark urine. Clinical evolution varied between one and eight days. Laboratorial findings in ill animals consisted of anemia, highly increased serum AST and GGT activities and urea and creatinine levels; whilst in the non symptomatic sheep and one goat revealed high and light GGT increase, respectively. Poultry litter analysis revealed levels of 449, 2610, 414 and 333 mg/kg of copper, iron, manganese and zinc, respectively. Analysis of copper levels by atomic absorption spectrometry in two sheep showed 730.1 and 1526.5 mg/kg in liver and 485.1 and 519.4 mg/kg in the kidneys. Treatment consisted of oral administration of 100 mg ammonium molibdate and 1 g sodium sulphate for five days. A total of 32 sheep died and necropsy of five animals showed deep jaundice, livers presenting a yellow to orange coloration, splenomegaly, hepatomegaly, blackish kidneys and dark urine. The microscopic examination of liver and kidneys showed degenerative and necrotic lesions.Discussion: This is the first report of copper poisoning in sheep from Rio Grande do Norte and Ceará, Northeastern Brazil. Even though it is not allowed in Brazil, the use of poultry litter for animal feeding still occurs and represents a great risk for copper poisoning in ruminants, especially sheep. The use of poultry litter should be discouraged to producers for feeding ruminants and requires more awareness of them that the use of this product is prohibited for this purpose. Clinical signs were typical of accumulative copper poisoning and the most evident changes in the serum biochemistry analyses were anemia and increased serum AST and GGT activities and urea and creatinine levels. Serum activity of GGT in a non symptomatic goat was slight increased, which is suggestive of an initial liver damage promoted by copper accumulation. Main pathological changes are associated to liver and kidney damage. Measurement of copper levels in liver and kidney fragments by atomic absorption spectrometry confirmed the diagnosis of poisoning by this element. Oral administration of ammonium molibdate associated to sodium sulphate was efficient for the treatment of poisoned sheep because both compounds reduce digestive absorption and increase excretion of copper. The absence of poisoning in goats fed the same poultry litter confirms that this species is more resistant than sheep to copper toxicity. Nevertheless further research must be carried out to test this sensibility. The use of poultry litter should be discouraged to producers for feeding ruminants and requires more awareness of them that the use of this product is prohibited for this purpose.Keywords: jaundice, hemoglobinuric nephrosis, poultry litter, small ruminants.

Relation of Copper Toxicosis in Dogs and Wilson Disease to the Appearance of a Small Copper Carrier (SCC) in Blood Plasma and Urine

The main route for copper excretion in mammals is via the bile, little exiting through urine. In Wilson disease, biliary excretion is prevented by lack of an active form of ATP7B, which permits copper entry into bile. This leads to copper accumulation and severe liver toxicosis. Dogs tend to have the same problem, with much higher liver copper levels than other mammals. Many die of liver copper toxicosis. As in Wilson disease, Atp7b‐/‐ mice excrete high levels of copper in urine. Excretion is in the form of a 1.5 kDa SCC. We now have evidence that the same SCC exists in the blood plasma of humans, mice and dogs. Normally, it is bound to larger proteins, but large amounts of free SCC occur in the plasma of Atp7b‐/‐ mice, and in some dogs. SCC can be released from larger proteins by EDTA and accounts for ~10% of the total copper in human plasma. SCC is also released by EDTA in plasma from dogs and mice, including those with “free” SCC. FPLC‐SEC preliminary data suggest SCC is riding on proteins &gt;200kDa. Dog ceruloplasmin is also unusual, eluting as a much larger protein, although of normal size in SDS‐PAGE. We conclude that SCC may offer an alternative route for copper excretion that can be mobilized in some mammals in response to liver copper accumulation, and that the study of dog plasma may reveal additional information about copper metabolism not detected as easily in other species.

The Exon 2 Deletion of the COMMD1 Causing Copper Toxicosis in Bedlington Terriers in Korea

This study was performed to survey prevalence of Copper metabolism domain containing 1 (COMMD1) mutation using molecular diagnostic method in a population of Bedlington terriers in Korea. COMMD1 gene (formerly MURR1) functions as a regulator of sodium transport and copper metabolism. The deletion of exon 2 of the COMMD1 gene causes copper toxicosis in Bedlington terriers. Bedlington terriers with this autosomal recessive disorder were shown to have the elevated liver copper levels due to genetic derangement in the biliary copper excretion pathway. DNA samples were extracted from whole blood collected from 257 Bedlington terriers (109 males, 148 females) of pet dog clubs in Korea. A multiplex PCR was carried out to detect of exon 2 deletion of COMMD1 gene. In this study, it was possible to know the existence and prevalence of exon 2 deletion of COMMD1 in Bedlington terriers in Korea. Of the 257 samples, 131 (51%) were wild type homozygous for the normal COMMD1 gene, 108 (42%) were heterozygous, having both normal and mutated copy of the COMMD1 gene. The eighteen (7%) were mutant type homozygous. The results of genetic analysis could help establish proper management strategy and selective breeding program to prevent COMMD1 mutation in Bedlington terriers in Korea.

Is it necessary to re-evaluate diagnostic criteria for Wilson disease in children?

The differential diagnosis of Wilson Disease (WD) is challenging, especially in children, because liver copper levels may also increase in other chronic liver diseases with bile stasis. The aim of this study is to determine urine and liver copper cut-off values to differentiate WD from other chronic liver diseases (non-WD, NWD) in children. Seventy-six patients participated in the study, 35 with WD and 41 with NWD. The two groups were divided into two subgroups according to the presence of cholestasis. At the time of diagnosis, age, sex, biochemical test results, serum ceruloplasmin, baseline 24-h urinary copper levels, liver biopsy histological findings, liver copper levels, and Child-Pugh scores were obtained from medical records. Copper content in liver tissue and copper levels in urine were measured by atomic absorption spectrometry. Cut-off values for differentiation of WD from NWD were determined by receiver operating characteristic (ROC) analysis. A liver copper cut-off value of 98 µg/g indicated WD with 91% sensitivity and 65.4% specificity (area under the curve =0.838, 95% CI: 0.749-0.927). A 24-h urinary copper cut-off value of 67.5 µg/24h indicated WD with 85% sensitivity and 71% specificity (area under the curve =0.843, 95% CI: 0.752-0.934). In this study of pediatric chronic liver disease patients, copper cut-off values for distinguishing WD differed substantially from those used for diagnosis. A larger scale study is warranted to re-evaluate liver copper and 24-h urinary copper cut-offs for children with suspected WD.

The Role of Copper and Vitamin A Deficiencies Leading to Neurological Signs in Captive Cheetahs (&amp;lt;i&amp;gt;Acinonyx jubatus&amp;lt;/i&amp;gt;) and Lions (&amp;lt;i&amp;gt;Panthera leo&amp;lt;/i&amp;gt;) in the United Arab Emirates

Neurological signs like ataxia and hind limb paresis have often been reported in cheetahs (Acinonyx jubatus), lions (Panthera leo) and snow leopards (Panthera unica). As a cause, copper and Vitamin A deficiencies have been discussed. Many cases were seen in cheetahs and lions in the United Arab Emirates (UAE) within the last years. The aim of this study was to find correlations between nutrition, serum, and tissue levels, focusing on copper and Vitamin A. Blood and tissue samples of affected and unaffected animals were analyzed at the Central Veterinary Research Laboratory in Dubai, UAE. Animals were split into three different groups (A, B and C) according to their diets. Minerals were determined in serum, tissue, food and water samples, and serum was additionally analyzed for Vitamin A and E. Liver, kidney and spinal cord samples were taken for histopathological investigations. Mean serum copper and liver copper levels of animals fed pure chicken muscle meat without supplements were significantly lower (0.41 ± 0.71 μM/L; 2.16 ± 0.95 ppm wet weight) than in animals fed a whole carcass prey diet (12.16 ± 3.42 μM/L; 16.01 ± 17.51 ppm wet weight) (p < 0.05). Serum Vitamin A and E levels were highest in animals fed whole carcass prey diets (1.85 ± 0.68; 27.31 ± 5.69 μM/L). Liver zinc concentrations were highest in animals fed pure chicken meat only (43.75 ± 16.48 ppm wet weight). In histopathology, demyelination of the spinal cord was found in all of the affected animals and most commonly when fed a diet based on poultry without supplements.

Levels of Chromium and Copper in Liver and Muscle Tissues of the Round Sardinella Sardinella aurita ( Valenciennes ) from the Oran Coastline , Algeria

The aim of the present study is to determine the levels of two heavy metals (Chromium and Copper) in the liver and muscle tissues of Sardinella aurita from the Oran coastline. Metal levels in fish samples were analyzed by using atomic absorption. 400 readings were carried out between May and October 2007. The order of heavy metal accumulation in the fish were in the order of magnitude as Cu > Cr. The average concentrations recorded in the fish, in this study, ranged, respectively, between 1.9 to 3.97 mg/kg for Copper, and between 0.07 to 0.12mg/kg for Chromium. The levels measured in liver tissue are higher than those reported for muscle tissue. The present study confirms that the round Sardinella Sardinella aurita (Valenciennes, 1847), from the Oran coastline, can bioaccumulate heavy metals from a polluted environment. The average concentrations of Cr (0.083±0.01mg/kg) and Cu (2.78±2.92, recorded in our study, were above FAO limiting standards for food fish.

Investigation of Iron Metabolism in Mice Expressing a Mutant Menke’s Copper Transporting ATPase (Atp7a) Protein with Diminished Activity (Brindled; MoBr/y)

During iron deficiency, perturbations in copper homeostasis have frequently been documented. Previous studies in iron-deprived rats demonstrated that enterocyte and hepatic copper levels increase and a copper transporter (the Menkes Copper ATPase; Atp7a) is induced in the duodenal epithelium in parallel to iron transport-related genes (e.g. Dmt1, Dcytb, Fpn1). Moreover, two ferroxidase proteins involved in iron homeostasis, hephaestin expressed in enterocytes and ceruloplasmin, produced and secreted into blood by the liver, are copper-dependent enzymes. We thus aimed to test the hypothesis that Atp7a function is important for the copper-related compensatory response of the intestinal epithelium to iron deficiency. Accordingly, iron homeostasis was studied for the first time in mice expressing a mutant Atp7a protein with minimal activity (Brindled [MoBr /y]). Mutant mice were rescued by perinatal copper injections, and, after a 7–8 week recovery period, were deprived of dietary iron for 3 weeks (along with WT littermates). Adult MoBr /y mice displayed copper-deficiency anemia but had normal iron status; in contrast, iron-deprived MoBr /y mice were iron deficient and more severely anemic with partial amelioration of the copper-deficient phenotype. Intestinal iron absorption in both genotypes (WT and MoBr /y) increased ∼3-fold when mice consumed a low-iron diet and ∼6-fold when mice were concurrently bled. WT mice exhibited no alterations in copper homeostasis in response to iron deprivation or phlebotomy. Conversely, upregulation of iron absorption was associated with increased enterocyte and liver copper levels and serum ferroxidase (ceruloplasmin) activity in MoBr /y mice, typifying the response to iron deprivation in many mammalian species. We thus speculate that a copper threshold exists that is necessary to allow appropriate regulate of iron absorption. In summary, MoBr /y mice were able to adequately regulate iron absorption, but unlike in WT mice, concurrent increases in enterocyte and liver copper levels and serum ferroxidase activity may have contributed to maintenance of iron homeostasis.

Does Dietary Copper Supplementation Enhance or Diminish PCB126 Toxicity in the Rodent Liver?

Copper is essential for the function of the mitochondrial electron transport chain and several antioxidant proteins. However, in its free form copper can participate in Fenton-like reactions that produce reactive hydroxyl radicals. Aryl-hydrocarbon receptor (AhR) agonists, including the most potent polychlorinated biphenyl (PCB) congener, 3,3',4,4',5-pentachlorobiphenyl (PCB126), increase copper levels in rodent livers. This is accompanied by biochemical and toxic changes. To assess the involvement of copper in PCB toxicity, male Sprague-Dawley rats were fed an AIN-93G diet with differing dietary copper levels: low (2 ppm), adequate (6 ppm), and high (10 ppm). After three weeks, rats from each group were given a single ip injection of corn oil (control), 1, or 5 μmol/kg body weight PCB126. Two weeks following injections, biochemical and morphological markers of hepatic toxicity, trace metal status, and hepatic gene expression of metalloproteins were evaluated. Increasing dietary copper was associated with elevated tissue levels of copper and ceruloplasmin. In the livers of PCB126-treated rats, the hallmark signs of AhR activation were present, including increased cytochrome P450 and lipid levels and decreased glutathione. In addition, a doubling of hepatic copper levels was seen, and overall metal homeostasis was disturbed, resulting in decreased hepatic selenium, manganese, zinc, and iron. Expression of key metalloproteins was either decreased (cytochrome c oxidase), unchanged (ceruloplasmin and CuZnSOD), or increased (tyrosinase and metallothioneins 1 and 2) with exposure to PCB126. Increases in metallothionein may contribute/reflect the increased copper seen. Alterations in dietary copper did not amplify or abrogate the hepatic toxicity of PCB126. PCB126 toxicity, i.e., oxidative stress and steatosis, is clearly associated with disturbed metal homeostasis. Understanding the mechanisms of this disturbance may provide tools to prevent liver toxicity by other AhR agonists.

Chronic copper intoxication causes impaired spatial memory and hippocampus copper elevation in wistar rats

Background: Animal models of copper toxicities rarely exhibit neurological impairments and increased brain copper accumulation impeding the development of novel therapeutic approaches to treat neurodegenerative diseases having high brain Cu content. Aims: The aim of this study was to investigate the effects of intraperitoneally injected copper lactate (0.15 mg Cu/100 g b.w.) daily for 90 days on copper and zinc levels in liver, kidney and hippocampus; expression of hepatic metallothionein-I and Atp7b gene, with metallothionein-III and acetylcholine esterase (AChE) gene in brain, biochemical parameters and neurobehavioral functions (by Morris water maze) of male wistar rats. Results: Copper administered animals exhibited significantly decreased serum AChE activity, increased hepatic metallothionein-I gene expression, impaired neuromuscular coordination and spatial memory compared to control rats. Copper intoxicated rats showed significant increase in liver, hippocampus and kidney tissues copper content (99.1, 73 and 74.9 % increase respectively), 40.7 % reduction in hepatic zinc content and interestingly, 77.1% increase in hippocampus zinc content with concomitant increase in copper and zinc levels in serum and urine compared to control rats. Massive copper deposition and copper associated protein in hepatocytes of copper intoxicated rats was substantiated by rhodanine and orcein stains respectively. Copper intoxicated rats demonstrated swelling and increase in number of astrocytes, degenerated neurons having pyknotic nuclei with copper deposition in choroid plexus. Conclusion: Present study shows the first evidence in vivo that chronic copper toxicity causes impaired spatial memory, swelling of astrocytes, decreased serum AChE activity,

Persistence of the efficacy of copper oxide wire particles against Haemonchus contortus in grazing South African goats

A study was conducted to examine the duration of anthelmintic effect of copper oxide wire particles (COWP) in grazing goats, as data for the persistence of efficacy of COWP in this host species is limited. Forty-eight indigenous male goats were infected naturally by grazing them on Haemonchus contortus-infected pasture. When the faecal egg count (FEC) in the goats was 3179±540 eggs per gram of faeces (mean±standard error), half the animals were treated with 4g COWP (day 0; mean live weight=25.5±0.8kg). Eight treated (COWP) and eight non-treated (CONTROL) goats were removed from the pasture on each of days 7, 28 and 56, maintained for 27 or 29 days in concrete pens and then humanely slaughtered for nematode recovery. Mean liver copper levels were in the high range in the goats removed from pasture at day 7 (treated: 191±19.7ppm; untreated: 120±19.7ppm; P=0.022), but had dropped to normal levels at days 28 and 56. The mean H. contortus burdens of the treated versus the non-treated goats were, respectively, 184±48 and 645±152 for the goats removed from pasture at day 7 (71% reduction; P=0.004), 207±42 and 331±156 at day 28 (37% reduction; P=0.945) and 336±89 and 225±53 at day 56 (−49% reduction; P=0.665). Weekly monitoring of FECs after treatment until slaughter indicated that the COWP-treated goats had lower FECs than the controls, the treatment main effect being significant at days 7, 28 and 56 (P<0.01). The day main effect and the treatment×day interaction were only significant for the goats removed from pasture at day 28 (P≤0.001). Packed cell volumes increased during the course of the experiment (day, P<0.001), but the treatment main effect was significant only for the goats removed from pasture at day 28 (CONTROL 28 d, 28.65±0.52%<COWP 28 d, 31.31±0.52%; P<0.001). No differences in live weight between groups were considered to be of any practical significance. The study indicated that persistence of efficacy of COWP is limited in goats, extending at most to 28 days after treatment. However, repeated COWP administration at three-month intervals may be safe, given that liver copper levels return to normal two to three months after COWP treatment.