Hepatic Copper Concentrations Research Articles

Improvement in Liver Pathology after 4 Months of d-Penicillamine in 5 Doberman Pinschers with Subclinical Hepatitis

Five female Doberman Pinschers with increased hepatic copper concentrations and persistent (3-4 years) subclinical hepatitis were treated with D-penicillamine for 4 months. Before and after treatment, the dogs underwent clinical, hematologic (red blood cell, white blood cell, and differential and thrombocyte counts), and clinical chemistry (creatinine, alkaline phosphatase, alanine aminotransferase, and total bile acid concentrations) examinations, and liver biopsies were examined histologically and their copper content measured quantitatively. No adverse effects were observed during treatment, and CBC and serum chemistry test results did not change. The mean liver copper concentration was 1,036 mg/kg dry matter before treatment and decreased to 407 mg/kg after treatment (P = .03). The copper concentrations had decreased (by between 134 and 1,135 mg/kg dry matter) in all of the dogs. The histopathologic appearance had improved or returned to normal in all 5 dogs. We conclude that D-penicillamine effectively reduced copper retention in these dogs and improved the histopathologic appearance of the lesions. However, because D-penicillamine has both copper-chelating and anti-inflammatory properties, it is not possible to draw conclusions on the etiology of this disease.

Wilson disease in septuagenarian siblings

Wilson Disease (WD) usually presents in the first decades of life, although rare patients have a later presentation. We report the clinical features, diagnostic evaluation, and outcome with treatment of two septuagenarian siblings evaluated as part of a research trial for treatment of neurological WD. The index case was a 72-year-old woman who suffered progressive neurological disability, then developed sub-fulminant liver failure. Her sibling was a 70-year-old man with minimal neurological symptoms and a mild depressive disorder. His liver biopsy revealed only steatosis and minimal fibrosis and an elevated hepatic copper content (671 mug/g dry weight liver). Molecular studies demonstrated compound heterozygosity for disease specific ATP7B mutations E1064A and H1069Q in both patients. Both individuals were treated with trientine and Zn followed by Zn maintenance therapy. Over the last 5 years, the clinical course stabilized and improved, although the index case recently died from bronchopneumonia. In conclusion, advanced age and different clinical presentations of these two subjects with identical ATP7B mutations raises the question of the degree of penetrance for these and other ATP7B mutations. Environmental and extragenic factors are pivotal determinants of disease phenotype. We suggest that WD must be considered at all ages in patients with hepatic disease, neurological disease, or psychiatric symptoms.

Recent clinical features of Wilson?s disease with hepatic presentation

We carried out this study to evaluate recent clinical features of Wilson's disease (WD) with hepatic presentation, especially in terms of age, degree of liver injury, and association with hepatocellular carcinoma (HCC). Sixteen patients with hepatic manifestations were diagnosed with WD in the period 1976-2003. We divided this period into two periods, "past" and "recent". The diagnosis was based on the presence of Kayser-Fleisher rings, low serum copper levels, low serum ceruloplasmin levels, increased urinary copper concentrations before or after D-penicillamine challenge, and increased hepatic copper concentrations. This retrospective study was done at Ehime University Hospital. Four patients, including a pair of siblings, had a family history of WD. Four patients had parental consanguinity. There were 6 patients aged over 40 years in the recent period, whereas no patients in the past period were over 40. Four patients had neurological manifestations. Ten patients had liver cirrhosis and 5 had chronic hepatitis. Two had fatty liver without obesity. All patients in the past period had liver cirrhosis. Three patients with liver cirrhosis were found to have HCC during the follow up. All patients were treated with either D-penicillamine or trientine chloride, or both. However, four patients had to discontinue these agents due to the side effects. Recently, the number of patients diagnosed with WD has been increasing, not only in terms of those with classical-type WD but also in terms of elderly patients or patients with non-cirrhotic liver injury such as fatty liver and chronic hepatitis. The various clinical features of WD should be recognized and particular attention should focus on HCC as a complication.

Vanadium induces apoptosis and modulates the expressions of metallothionein, Ki‐67 nuclear antigen, and p53 during 2‐acetylaminofluorene‐induced rat liver preneoplasia

Our previous studies have shown that vanadium, a dietary micronutrient, has an inhibitory effect against experimentally induced rat hepatocarcinogenesis. In this study, we evaluated the role of vanadium on some potential protein expression markers of carcinogenesis, such as metallothionein (MT), an intracellular metal-binding protein linked with cell proliferation and apoptosis, Ki-67 nuclear antigen, and p53 tumor suppressor during 2-acetylaminofluorene (2-AAF)-induced (0.05% in basal diet) rat liver preneoplasia. In a short-term regimen, supplementation of vanadium at a dose of 0.5 ppm effectively suppressed the formation of DNA 'comets' (29.55%; P < 0.02), thereby indicating its nongenotoxicity at this particular dose. Vanadium administration throughout the study reduced relative liver weight (RLW), nodular incidence (57.15%), total number, and multiplicity (48.45%) with restoration of hepatic zinc (Zn), magnesium (Mg), selenium (Se), copper (Cu), iron (Fe), and calcium (Ca) contents when compared to the carcinogen control. Moreover, treatment with vanadium significantly abated the expressions of MT and Ki-67, studied at four sequential time points. An increased immunopositivity of p53 protein (1.03 +/- 0.23%; P < 0.02) was found in vanadium-treated rat liver with an elevated apoptotic-labeling index (AI; P < 0.001) as documented by TUNEL assay. Furthermore, a positive correlation between MT expression and Ki-67 labeling along with a strong negative correlation between MT immunoreactivity and AI (r = -0.9000, P = 0.0004 at week 24) at various time intervals suggest that, vanadium-mediated suppression of MT and Ki-67 expressions may be associated with induction of apoptosis. The results thus provide evidence for the first time in support of the potential role of vanadium on induction of p53 and apoptosis with concurrent suppression of MT and Ki-67 in order to have an understanding, in part, of the chemopreventive mechanism of this trace element in limiting neoplastic transformation in a defined model of experimental rat hepatocarcinogenesis.

Suppressive effects of dietary D-penicillamine on the gene expression of hepatic metallothionein-1 and protein level of hepatic metallothionein in Long-Evans Cinnamon rats

We investigated the effects of dietary D-penicillamine on copper and zinc contents in the liver, serum, and urine of Long-Evans Cinnamon (LEC) rats, as well as the gene expression of hepatic metallothionein-1 and hepatic metallothionein protein level in these animals. Young male LEC and Fischer rats were fed a basal or D-penicillamine (5 g D-penicillamine/kg diet) diet for 16 days. The urinary output of copper and zinc was markedly increased by the D-penicillamine diet, and the copper and zinc contents in the liver were significantly decreased in LEC rats. Excess copper accumulation in the liver induced hepatic metallothionein in LEC rats, but the D-penicillamine diet depressed the level of hepatic metallothionein-1 mRNA as well as the hepatic metallothionein protein level. When fed the basal diet, serum ornithine carbamoyltransferase activity was higher in LEC rats than in Fischer rats, but a significant decrease in serum ornithine carbamoyltransferase activity rats was observed in LEC rats on feeding the D-penicillamine diet. These results indicated that dietary D-penicillamine significantly reduced excess copper and zinc in the livers of LEC rats through urinary excretion, and the decrease in hepatic copper and zinc contents caused a decrease in the levels of metallothionein-1 mRNA and metallothionein protein in LEC rats.

Copper-induced oxidative stress in three-spined stickleback: relationship with hepatic metal levels

The aim of this study was to characterise biomarker responses in three-spined sticklebacks exposed to copper. For this purpose, adult sticklebacks were exposed for 3 weeks to copper sulphate at 0, 25, 100 and 200 μg L −1 as Cu. At days 4, 8, 12 and 21, several parameters were measured including liver, gonad and spleen somatic indexes, hepatic biomarkers (catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (GSH), glutathione- S-transferase (GST) and 7-ethoxyresorufin-O-deethylase (EROD)) and hepatic copper and zinc concentrations. Copper induced a rapid and transient increase of antioxidant enzymes and a depletion of glutathione content during the first 8 days of exposure. Significant copper and zinc accumulation in fish liver were observed for the two higher exposure concentrations after 8 and 12 days, respectively. This study showed that copper induced an oxidative stress in fish liver before significant metal accumulation in the liver could be detected, suggesting the involvement of differential mechanisms in copper uptake and metabolism. Three-spined stickleback appears to be a sensitive model to study oxidative stress induced by metals.

Inhibitory effects of trientine, a copper-chelating agent, on induction of DNA strand breaks in hepatic cells of Long–Evans Cinnamon rats

Effects of treatment with trientine, a specific copper-chelating agent, on accumulation of copper and induction of DNA strand breaks were investigated in Long-Evans Cinnamon (LEC) rats, an animal model for human Wilson's disease. Copper accumulated in the livers of LEC rats in an age-dependent manner from 4 to 13 weeks of age. When LEC rats were treated with trientine from 10 weeks of age, hepatic copper contents did not increase and were maintained at the same levels as those in 10-week-old LEC rats. When the amounts of DNA single-strand breaks (SSBs) were estimated by a comet assay, SSBs of DNA were induced in a substantial population of LEC rat hepatic cells around 8 weeks of age and the amounts of SSBs increased in an age-dependent manner from 8 to 15 weeks of age. When LEC rats were treated with trientine from 10 weeks of age, the observed number of cells with DNA damage decreased dramatically, suggesting that induction of SSBs of DNA was inhibited and/or SSBs were repaired during the period of treatment with trientine. The results show that treatment of LEC rats with trientine decreases the number of DNA strand breaks observed, although copper contents remain high in the liver.

Wilson disease.

Wilson disease (WD) is an autosomal recessive disorder of copper metabolism. Since daily copper intake exceeds the body's requirements, effective means of excreting excess copper are essential. These are accomplished by ATP7B, a new member of the cation-transporting p-type ATPase family, which is mainly expressed in the liver and mediates both copper secretion into plasma (coupled with ceruloplasmin synthesis) and its excretion into bile. Thus far, more than 200 mutations of the WD gene have been detected, causing impairment of ATP7B function and, ultimately, copper accumulation. Excess copper, however, induces free-radical reactions and lipid peroxidation. Resultant liver damage leads to steatosis, inflammation, cirrhosis, and, occasionally, fulminant liver failure. The diagnosis of WD is commonly made on the basis of typical clinical and laboratory findings, including low serum ceruloplasmin, increased urinary copper excretion, and increased hepatic copper content. Since liver morphology is non-specific, and copper histochemistry may lead to both false-negative and false-positive results, the pathologist usually only suspects the disease or assists in its confirmation. Although the value of molecular genetic testing is limited due to the high number of possible gene mutations, polymerase chain reaction may be useful for the evaluation of family members of homozygous index patients.

P0207 EVALUATION OF NON-INVASIVE TESTS??? SENSITIVITY IN DIAGNOSIS OF WILSON DISEASE IN CHILDREN

Introduction: Wilson disease (WD) is an autosomal recessive disorder disturbing copper metabolism. The responsible gene (ATP7B) coding membrane-bound copper-binding proteine is situated on chromosome 13 q14–q21. The diagnosis of WD depends on combination of non-ivasive (ceruloplasmin, serum copper, 24h urinary copper excretion), invasive (hepatic copper) and genetic (mutation analysis) findings. Methods: 27 patients (aged 9–18, 16M, 11F) with confirmed WD were involved to the study. In 23 of them the diagnosis was confirmed by identification of two mutations of ATP7B (H1069Q was the most predominant – 42%). In 4 outstanding WD was confirmed by increased (> 250 mg/g dry weight) hepatic copper content. All but one (with hepatoneurological symptoms) suffer from liver symptoms. Results: Ceruloplasmin <20 [mg/dl], serum copper <70 [mg/dl], 24h urinary copper excretion >100 [mg/24h] were regarded as values characteristic for Wilson’s disease. The results of non-invasive tests were as follows [mean±SD; 95% CI]: ceruloplasmin 9,12±8,14; 5,60–12,64 [mg/dl], serum copper 34,69±19,83; 26,11–43,27[mg/dl], 24h urinary copper excretion 267,95±452,77; 67,20–468,70 [mg/24h]. The sensitivity of these tests were: 87% (20/23) for ceruloplasmin, 96% (22/23) for serum total copper and 73% (16/22) for 24h urinary copper excretion. None of our patients had three tests within normal ranges and one had two of three tests within normal ranges. Conclusion: 1. Serum total copper seems to have the highest sensitivity. 2. The sensivities of ceruloplasmin and 24h urinary copper excretion are too low to be used as screening tests 3. Evaluation of all three non-invasive tests in differential diagnosis of liver disease is recommended.

Efeito da administração parenteral de cobre sobre o ganho de peso, eritrograma e parênquimas, hepático e renal, em bovinos mestiços (Zebu x Europeu) confinados

This study evaluated the effect of the parenteral administration of etilenodinitrile tetracetate of calcium and copper on the weight gain, eritrograma and the hepatic and renal parenchymas in confined bovines, using 200 animals, distributed in four groups (G) of 50 (I, II, III and IV). The groups I and II were constituted by bovines with the age of 24 months and the groups III and IV by animals of 12 months, which GI was given 100mg of active copper and GIII 75mg, both by subcutaneous way by the beginning of the study. The other groups were used as the control. The weight measure of the animals of all groups was done from 28 to 28 days during the 112 days of confinement. It was done the eritrograma in ten bovines of each group and the dosage of the hepatic and renal concentration of copper and the hystological evaluation, in the animals of the groups I and II. The values found on the eritrograma showed normality and the hepatic and renal concentrations of copper in bovines with the age of 24 months showed depletion levels. It was not verified differences about the hystological evaluation between the groups I and II. There was significant difference on the weight gain media between the groups I and II. On the groups III and IV, it was not observed difference over the weight gain media.

Hepatic Cobalt and Copper Levels in Lambs in Norway

Cobalt and copper concentrations were measured in 599 lamb livers collected at slaughter from 58 sheep flocks in 6 different parts of Norway in 1993. Information about pasture, additional feeding and mineral supplements in the flocks was obtained through a questionnaire. Average hepatic levels of cobalt in the lamb flocks varied from <0.003 to 0.22 μg/g ww, and of copper from 5 to 240 μg/g ww. Flocks with deficient or marginal cobalt status were found in all parts of southern Norway, but primarily in the west and south-west. Some flocks with marginal copper status were found in the south-west, while flocks with signs of excessive hepatic copper concentrations were found mainly in inner parts of central and northern Norway. Hepatic copper concentrations were significantly higher in lambs that had grazed mountain pastures than in those that had grazed lowland pastures in the summer.

Effect of restricted feed intake and addition of vitamins B2 and B6 and folic Acid on the liver concentration of zinc and copper in rats.

The aim of the present study was to investigate the influence of nutritional deficiency and dietary addition of vitamins (B2, B6, and folate) on hepatic concentration of zinc and copper in rats. The experiment was performed on 260 growing male Wistar rats divided into 13 groups. Animals of 11 groups were fed isocaloric diets (14.7 MJ/kg) in which the 20% of energy was derived from protein. Another two groups of rats were offered diets with 9% or 4.5% of energy originating from protein. Animals of both mentioned groups and of the control group (20% of energy from protein) were offered diets ad libitum. The other 10 groups were offered 50% and 30% of the amount consumed in the control group. Eight groups, from those 10 restricted ones, were differentiated by dietary addition of vitamins B2 and B6 and folate (300% addition). Restricted feed intake did not affect the liver zinc concentration but significantly increased the copper concentration. The addition of vitamin B6 decreased the liver Zn concentration. The highest liver Cu concentration was noted in rats offered restricted diets to only 30% of intake in the control group and high in vitamin B2 and in rats supplemented with all of studied vitamins together. It suggests that vitamin B2 had the strongest impact on liver Cu concentration in rats fed restricted diets.

Inherited canine copper toxicosis in Australian Bedlington Terriers

Inherited copper toxicosis in Bedlington Terriers (CTBT) is a copper associated hepatopathy caused by an autosomal recessive genetic defect of gene involving copper metabolism. To compare clinical and histopathological findings with previous reports and to expand our knowledge for future genetic studies, 18 terriers were clinically and histopathologically examined in this study. Pedigree information and dietary history were obtained from the owners before a thorough clinical examination was undertaken. Following the examination, a blood sample was collected for haematology, biochemistry and genetic analysis and a urine sample for urinalysis. Seven dogs were also liver biopsied for histopathology, histochemistry and electron microscopy. In this study, plasma alanine transaminase (ALT) activity was highly concordant with DNA marker test results and was the most reliable and sensitive biochemical test measured. Also clinical and biochemical copper toxicosisaffected states were noticed in a genotyped carrier dog. Histopathological and electron microscopy findings showed that the severity of the lesion was more closely correlated to the presence of clinical signs than to hepatic copper concentration. In addition, the involvement of apoptosis and p53 gene was observed in electron microscopy. The general findings related to CT-BT in this study was similar to those previously reported except few differences in histopathology and electron microscopy.

Wilson disease

Wilson disease

Current concepts in the treatment of canine chronic hepatitis

Chronic hepatitis is a common disorder in dogs seen by general practitioners. Several new drugs have been marketed for treating this disease. Unfortunately, there are few controlled studies that examine the efficacy of these medications for the treatment of canine chronic hepatitis. A rational therapeutic approach can be implemented based on histopathologic findings of a liver biopsy. A liver biopsy is essential for establishing a definitive diagnosis and guiding the optimal therapy. The biopsy allows characterizing the inflammatory process, quantitating hepatic copper concentrations, and determining if fibrosis is present. Copper associated hepatopathy is treated with zinc and copper chelators. Idiopathic chronic hepatitis is thought to be immune mediated. The treatment of idiopathic chronic hepatitis consists of controlling inflammation (prednisone, azathioprine), reversing fibrosis (colchicine), and protecting against oxidant damage (vitamin E, ursodeoxycholic acid, S-adenosylmethionine). The prognosis for chronic hepatitis is quite variable. Dogs with end-stage disease have a poor prognosis, while dogs diagnosed earlier can have a mean survival of years. Early diagnosis and intervention are key to the successful treatment of dogs with chronic hepatitis.

Cholesterol and copper in the liver of rabbit inbred strains with differences in dietary cholesterol response

In order to investigate whether cholesterol intake influences the hepatic copper content of rabbits, we compared the hepatic copper content of two rabbit inbred strains after feeding the animals a control or a cholesterol-rich diet. One strain was not reactive to dietary cholesterol (IIIVO/JU), whereas the other strain was reactive to dietary cholesterol (AX/JU). The coefficient of inbreeding (F) >0.95 for both strains. Dietary cholesterol-reactive rabbits when compared with their non-reactive counterparts had a higher hepatic copper content. The consumption of a hypercholesterolemic diet decreased liver copper concentration (expressed in μg/g dry weight) in both strains of rabbits, which was (in part) due to dietary-induced hepatomegaly. A decrease in the absolute hepatic copper content was found only in the dietary cholesterol-reactive inbred strain. It is discussed that differences in glucocorticoid levels may be responsible for the strain difference in liver copper content. The cholesterol effect on the hepatic copper content in the reactive strain might be caused by an increased bilirubin secretion.

Elements in autopsy liver tissue samples from Greenlandic Inuit and Danes. IV. Copper measured by X-ray fluorescence spectrometry

Objective: To measure the content of copper (Cu) in liver tissue samples from Greenlandic Inuit and compare the results with those obtained in Caucasian Danes. Materials and methods: Normal liver tissue samples were obtained at autopsy from 50 Inuit (27 men, 23 women) with a median age of 61 years (range 23 - 83) and from 74 Danes (44 men, 30 women) with a median age of 60 years (range 15 - 87). Total liver copper content was measured by X-ray fluorescence spectrometry. Results: The content of copper given as median and 5 - 95 percentile was in Inuit 0.2982 mmol/kg dry liver (0.1810 -0.6531), and in Danes 0.3777 mmol/kg dry liver (0.1889 - 0.7550) (p = 0.02). This difference was due to a higher median liver copper content in Danish men compared with Danish women (p = 0.04), Inuit men (p = 0.005) and Inuit women (p = 0.05), and was based solely on the 3 highest liver copper values in Danish men. Inuit displayed no gender difference concerning liver copper content. There was no correlation between liver copper content and age either in Inuit or in Danes. In Inuit, the median hepatic copper index (liver copper content divided by age) was 0.0045 in men and 0.0066 in women (p = 0. 14). There was an inverse correlation between hepatic copper index and age in the entire Inuit series (r s = -0.63, p < 0.0001). In Danes, the median hepatic copper index was 0.0082 in men and 0.0046 in women (p = 0.005). There was an inverse correlation between hepatic copper index and age in the entire Danish series (r s = -0.76, p < 0.0001). Conclusion: There is no evidence to suggest that the prevalence of copper deficiency or copper overload in adults differ in Inuit and Danes. Inuit have a hepatic copper content, which is similar to that found in Caucasian Danes and other populations from the Western societies.

Liver copper content of rats hypo- or hyperresponsive to dietary cholesterol

The question addressed is whether cholesterol intake reduces the hepatic copper content in rats. For this purpose we have compared the hepatic copper content of two selected rat inbred strains after feeding the animals a control or a high fat, high cholesterol diet. One strain was dietary cholesterol resistant (SHR/OlaIpcv), whereas the other strain was susceptible to dietary cholesterol (BN- Lx/Cub). Dietary cholesterol-susceptible rats have a lower baseline hepatic copper content when compared with their resistant counterparts. The consumption of a hypercholesterolemic diet decreased the liver copper concentration (expressed in μg/g dry weight) to about the same extent in both strains. However, dietary cholesterol did not reduce the absolute (expressed as μg/whole liver) and relative (expressed as μg/whole liver/100 g body weight) copper store of rats. The decrease of liver copper concentration after the high fat, high cholesterol diet is probably not caused by a decrease in whole hepatic copper content, but rather due to dietary-induced hepatomegaly.

Copper‐Associated Liver Disease in Dalmatians: A Review of 10 Dogs (1998–2001)

This retrospective study summarizes 10 Dalmatians suspected of having hepatic copper toxicosis. Hepatic copper toxicosis can result from either a primary metabolic defect in hepatic copper metabolism or from altered hepatic biliary excretion of copper. An inherited copper‐associated hepatopathy has been documented in Bedlington Terriers, and there is evidence for familial copper‐associated liver disease in West Highland White (WHW) Terriers and Skye Terriers. Nine of the 10 Dalmatians in this study presented for gastrointestinal clinical signs, including anorexia and vomiting. All animals had increased alanine aminotransferase (ALT) enzyme activity, and 9 of 10 had increased alkaline phosphatase (ALP) enzyme activity. The relative increase in ALT activity was much greater than the relative increase in ALP activity, suggesting a predominantly hepatocellular rather than cho‐lestatic liver disease. The mean hepatic copper concentration for 9 Dalmatians was 3,197 μg/g dry weight liver (dwl) (normal, &lt;450 μg/g). In 5 of these 9 dogs, hepatic copper concentrations exceeded 2,000 μg/g dwl. Necroinflammatory alterations associated with copper‐laden parenchymal cells were the notable histopathologic finding. The inflammatory infiltrate was either primarily lymphocytic or neutrophilic. Morphologic features of cholestasis generally were not prominent except in those dogs with severe pathology. These findings lend support to the hypothesis that a primary metabolic defect in hepatic copper metabolism occurs in the Dalmatian breed. The mechanism and genetic basis of this condition require further study.

Early cell transplantation in lec rats modeling Wilson's disease eliminates hepatic copper with reversal of liver disease

Background & Aims:The Long-Evans Cinnamon (LEC) rat is an excellent model of Wilson's disease with impaired copper excretion, hypoceruloplasminemia, and copper toxicosis. We hypothesized that early hepatocyte transplantation would improve copper excretion and liver disease in Wilson's disease. Methods: Normal syngeneic Long-Evans Agouti rat hepatocytes were transplanted intrasplenically into 2-week-old LEC rats. Untreated LEC pups were controls. Liver repopulation was shown by changes in serum ceruloplasmin, hepatic atp7b messenger RNA. and bile and liver copper levels. Histologic analysis of the liver was performed.Results:Significant copper accumulation and liver disease were observed in 5-month-old LEC rats, with occasional treated rats showing increased bile copper excretion. The liver was repopulated extensively in 10 of 14 treated LEC rats (71%) 20 months after cell transplantation. In these 10 rats, hepatic copper content was virtually normal in 6 rats (53 ± 12 μg/g liver) and substantially less in 4 others (270 ± 35 μg/g) compared with elevated liver copper levels in untreated LEC rats (1090 ± 253 μg/g) (P < 0.001). Changes in serum ceruloplasmin levels, bile copper excretion capacity, and liver histology were in concordance with decreases in liver copper levels.Conclusions:Transplanted cells proliferated subsequent to the onset of liver injury, and the liver was repopulated over an extended period. Cell transplantation eventually restored copper homeostasis and reversed liver disease without hepatic preconditioning in LEC rats.