Liver Copper Concentrations Research Articles

Comparison of some meat quality and liver characteristics in Muscovy and mule ducks.

The aim of the study was to compare Muscovy ducks and mule ducks for proximate analysis, colour attributes, sensory properties of the meat, and concentration of some minerals in the meat and liver, with consideration of the effect of sex on the analysed traits. The study used 46 Muscovy and 44 mule ducks. At the end of the rearing period, 40 birds (10 males and 10 females of each genotype) were selected for slaughter. Meat and liver samples were collected from the slaughtered birds to determine quality traits. The analysed ducks of different genotypes differed significantly in the water content, sodium content, and redness of breast muscles; in the water, fat, and zinc content of leg muscles; and in the sodium, iron, and copper content of liver. Regardless of genotype, males had a higher sodium content in breast muscles; contained more protein, sodium, phosphorus, and magnesium and less potassium and copper in leg muscles; and had a significantly higher content of sodium, potassium, phosphorus, magnesium, zinc, iron, and copper in liver compared to females. The genotype–sex interaction was significant for the sodium and potassium content of breast muscles, for the water, protein, sodium, potassium, phosphorus, magnesium, and copper content of leg muscles, and for the copper content of liver. Our study provided information about differences in the quality of meat and liver in Muscovy and mule ducks.

Alterations in Copper Homeostasis in Rabbit Model of Atherosclerosis

Copper concentrations decrease in atherosclerotic tissues. The present study was undertaken to determine overall alterations in copper homeostasis in the pathogenesis of atherosclerosis. New Zealand white rabbits, aged 10‐weeks‐old, were fed a rodent chow diet containing 1% (w/w) cholesterol or the same diet without cholesterol as control. Twelve weeks after respective diet feeding, the animals were euthanatized by sodium pentobarbital. Rabbits fed cholesterol‐containing diet developed hyperlipemic as defined by a significant increase in the total plasma cholesterol and atherosclerotic lesions in aorta. Copper concentrations in the atherosclerotic tissues were significantly decreased while serum copper concentrations were increased in the hyperlipemic animals in comparison with controls. A detailed analysis revealed that copper concentration in abdominal aorta of the hyperlipemic rabbits was negatively correlated with the severity of the atherosclerotic lesion, stenosis of the lumen area, and collagen deposition. In addition, copper concentrations in the kidney of hyperlipemic rabbits were increased, but in the heart, liver, spleen and lung were decreased. These results suggest that in the pathogenesis of atherosclerosis, changes in copper homeostasis are not only observable in the atherosclerotic tissues, but also in multiple organ systems. These changes would be additive to atherosclerosis as a high risk of cardiovascular disease in humans.Support or Funding InformationWest China Hospital, Sichuan University(A) The percentage of the aortae staining positively for lipid(Longitudinal of suprarenal abdominal aortae was stained with oil red O). Sections of infrarenal abdominal aortae were stained with EVG for microscopic quantification of lesion areas, (B) the atherosclerotic lesion area and (C) the percent of stenosis of lumen area. Collagen fibers in the lesions were assessed by picrosirius red staining, the percent of Collagen in lesion area. (*P < 0.05)Figure 1The concentration of copper in tissues and plasma.(A) cardiac copper concentration; (B) liver copper concentration; (C) spleen copper concentration; (D) lung copper concentration; (E) kidney copper concentration; (F) duodenum copper concentration; (G) abdominal aortic copper; (H) plasma copper concentration. (*P < 0.05)Figure 2

Metal contamination of river otters in North Carolina.

Aquatic apex predators are vulnerable to environmental contaminants due to biomagnification. North American river otter (Lontra canadensis) populations should be closely monitored across their range due to point and nonpoint pollution sources. Nonetheless, no information exists on environmental contaminants in the North Carolina otter population. Metals and metalloids occur naturally across the landscape, are essential for cellular function, and become toxic when concentrated unnaturally. We conducted our study across the three Furbearer Management Units (FMU) and 14 river basins of North Carolina. We determined the concentrations of arsenic, cadmium, calcium, cobalt, copper, iron, lead, magnesium, manganese, mercury, molybdenum, selenium, thallium, and zinc in liver and kidney samples from 317 otters harvested from 2009 to 2016. Arsenic, lead, and thallium samples were tested at levels below the limit of detection. With the exception of cadmium, we detected all other elements at higher levels in the liver compared with the kidney. Specifically, cadmium, cobalt, copper, iron, magnesium, manganese, mercury, molybdenum, and zinc levels differed by tissue type analyzed. Most element concentrations remained stable or increased with otter age. We detected higher levels of mercury and selenium in the Lower Pee Dee and Cape Fear river basins. River basins within the Mountain FMU were higher in cadmium, copper, iron, lead, and zinc, whereas the Coastal Plain FMU was lower in cobalt and manganese. None of the elements occurred at toxic levels. Our research establishes baseline concentration levels for North Carolina, which will benefit future monitoring efforts and provide insight into future changes in the otter population.

PSIII-32 The influence of liver copper concentration on copper homeostatic liver proteins in beef cattle

Abstract The objective of this experiment was to investigate the influence of liver Cu concentrations on the relative abundance of liver Cu homeostatic proteins in beef cattle. Archived liver biopsy samples were selected based on Cu concentrations (n = 4 samples 21.7±1.35 mg Cu/kg DM-deficient; and n = 4 samples 73.3 ±10.7 mg Cu/kg DM-adequate). Liver samples were obtained from a subset of multiparous beef cows receiving a forage-based diet with no supplemental Cu (basal diet 6.25 mg Cu/kg DM) or 10 mg Cu/kg DM total diet (Cu supplemented as CuSO4·5H2O) for 99 d. Liver proteins were identified using mass spectrometry, normalized, and relative abundance determined using Scaffold software. A total of 895 identical proteins were identified in each sample and relative abundance of each Cu specific homeostatic protein (n = 13) was recorded. Data were analyzed as a randomized complete block design using R software. Copper homeostatic liver proteins identified were: aldehyde dehydrogenase, apolipoprotein A-1, betaine homocysteine methyltransferase, carbonic anhydrase II, Cu chaperone for superoxide dismutase, Cu transport protein, cytochrome c oxidase Cu chaperone, extracellular superoxide dismutase, flavin reductase, glutamate dehydrogenase, glutathione synthetase, protein disulphide isomerase A3, and Cu-zinc superoxide dismutase. By design, liver Cu concentrations were greater (P < 0.05) in Cu adequate vs. Cu deficient liver samples. Copper deficient liver samples had greater (P < 0.05) relative abundance of glutathione synthetase compared to Cu adequate liver samples. The relative abundance of all other Cu homeostatic liver proteins identified were similar (P > 0.05) across Cu concentrations. These data suggest that deficient and adequate liver Cu concentrations ranging from 16.0 to 109.0 mg Cu/kg DM have minimal impact on the relative abundance of Cu homeostatic proteins in beef cattle. Further investigation is needed to determine if liver Cu concentration influences Cu homeostatic protein function.

Influence of a mineral supplement containing calcium, phosphorus and micronutrients on intake, digestibility, performance and mineral status of young Nellore bulls in a feedlot

We measured nutrient intake and digestibility, performance and mineral status of young Nellore bulls fed three diets with and without supplementation comprising calcium (Ca), phosphorus (P) and micro-minerals during the growing and finishing phases. Five bulls were slaughtered at the beginning of the experiment to measure initial mineral status. Forty-two young Nellore bulls (initial bodyweight (mean ± s.d.) 270.4 ± 36.6 kg, age 8 months) were assigned for 125 days to one of six treatments: sugarcane as a roughage source plus a concentrate based on soybean meal and soybean hull with (SH100) and without (SH0) supplementation; sugarcane as a roughage source plus a concentrate based on soybean meal and ground maize with (SC100) and without (SC0) supplementation; and maize silage as a roughage source plus a concentrate based on soybean meal and maize with (CS100) and without (CS0) supplementation. The experiment was conducted as a completely randomised design with a 3 × 2 factorial arrangement of treatments. Nutrient intake and digestibility, bone and serum parameters related to Ca and P metabolism, and liver mineral concentrations were measured. Nutrient intake, digestibility and performance were not affected (P > 0.10) by the mineral factor. Water intake was higher (P < 0.05) with SH100 than with all other diets. Rib-bone breaking strength and density according to dual-energy X-ray absorptiometry were reduced (P < 0.04) in the absence of supplementation. Metatarsus parameters were not affected (P > 0.10). Liver copper (Cu) content was reduced (P < 0.01) in diets without supplementation. In conclusion, the short-term absence of mineral supplementation did not influence intake and digestibility. However, depending on diet type, absence of Cu may reduce the hepatic Cu content, and a low Ca:P ratio may stimulate bone reabsorption without compromising performance. Therefore, under our conditions, mineral requirements are overestimated by nutritional systems.

Dietary copper requirement of fingerlingHeteropneustes fossilisfor formulating copper‐balanced commercial feeds

Dietary copper requirement of Heteropneustes fossilis (6.74 ± 0.03 g) was determined by feeding purified diets containing same protein (400 g/kg) and gross energy (17.89 kJ/g) but different levels of copper for 12 weeks. Graded amount of CuSO₄.5H₂O (0, 1.96, 3.93, 5.89, 7.86, 9.82, 11.79 mg/kg) was supplemented to basal diet to attain desired dietary copper levels (0, 0.5, 1.0, 1.5, 2.0, 2.5 and 3.0 mg/kg). Analysed dietary copper concentrations were 4.28, 4.63, 5.28, 5.70, 6.19 and 6.69 mg/kg. Absolute weight gain, feed conversion ratio and protein gain improved with the increasing levels of dietary copper up to 5.28 mg/kg. Further inclusion of copper at a level of 5.70 mg/kg did not improve the above parameters. Significantly higher (p < .05) plasma ceruloplasmin, liver copper‐zinc superoxide dismutase, catalase activities and lower thiobarbituric acid reactive substances were evident in fish receiving diets with 5.28 and 5.70 mg/kg copper compared to other groups. Whole body and liver copper concentrations increased significantly (p < .05) with increasing dietary copper levels. Quadratic regression analysis of absolute weight gain, feed conversion ratio, protein gain and broken‐line regression analysis of plasma ceruloplasmin activity and liver TBARS value against the variable dietary copper levels depicted the dietary copper requirements for fingerling H. fossilis in the range of 5.24–5.68 mg/kg.

Effect of copper exposure and recovery period in reared Diplodus sargus.

The aquaculture growth can be followed by the occurrence of more and new pathogenic agents, since the production leads to higher fish densities in confined areas more appropriate to the appearance and propagation of pathologies. Copper sulfate has been widely used in preventing and controlling fish parasites. The objective of this study is to investigate the effects of copper treatments in the fish tissues (bioaccumulation and histological changes in different organs), mortality and evaluate what happens during the recovery period. White sea bream (Diplodus sargus) were exposed to copper sulfate (0.25 and 0.5 mg L-1) during 60 days followed with a 75-day recovery period. The results showed that the concentration of copper in fish liver was significantly higher in the 0.5 mg L-1 treatment than in the 0.25 mg L-1 treatment. Conversely, copper load in the muscle did not differ significantly between treatments and control. Copper levels in muscle, and especially in liver, increased during copper exposure (up to 60 days). In summary, at higher concentrations copper sulfate treatment (0.5 mg L-1) might be toxic to fish, which showed histological alterations and copper accumulation in their tissues, mainly in the liver. Nevertheless, individuals returned to their original state after a 75-day recovery period and the tested copper concentrations does not represents risk for food safety.

Long-term effects of chromium on morphological and immunological parameters of Wistar rats

Hexavalent chromium raises high concern because of its wide industrial applications and reported toxicity. Long-term (135 days) oral exposure of Wistar rats to chromium in the form of K2Cr2O7 (exposed group~20 mg/kg/day) led to a decrease in thymus mass and thymocytes' number and caused structural and functional changes in the lymph nodes and spleen, namely lymphoreticular hyperplasia and plasmocytic macrophage transformation. Programmed cell death was increased in both thymocytes and splenocytes and decreased in lymphocytes in the T-zones of spleen and lymph nodes. Moreover, Cr (VI) administration decreased myeloid cells' and neutrophils' number, while it increased lymphoid and erythroid cells' number in bone marrow. Cr (VI) immune system effects seem to be related to oxidative stress induction, as depicted by the increased levels of diene conjugates and malondialdehyde in the spleen and liver and by the decreased activity of catalase and superoxide dismutase in rats’ erythrocytes. In addition, exposure to Cr (VI) decreased copper, nickel and iron concentrations in blood and liver, while Cr levels in blood, spleen and liver were increased, as expected. The observed changes in the series of immunological parameters studied contribute to the development of new approaches for the prevention of low level Cr exposure toxicity.

Copper, Iron, Manganese, Zinc, Cobalt, Arsenic, Cadmium, Chrome, and Lead Concentrations in Liver and Muscle in Iranian Camel (Camelus dromedarius).

Camel meat as healthy food has received much attention for human consumption. In the present study, liver and muscle from 60 camels (Camelus dromedarius) including 26 males and 34 females were sampled to determine the concentration of As, Cd, Pb, Cr, Cu, Fe, Mn, Zn, and Co using the inductively coupled plasma-optical emission spectrometry (ICP-OES). Based on the obtained results, the average content of essential or toxic elements in the liver and muscle was as follows: 111.2 ± 26.51, 38.57 ± 7.97 (Zn), 3.28 ± 0.79, 2.12 ± 0.49 (Cu), 76.98 ± 14.20, 59.34 ± 11.81 (Fe), 0.87 ± 0.22, 0.48 ± 0.12 (Mn), 0.52 ± 0.27, 0.03 ± 0.01 (Co), ND ± 0.008, 0.012 ± 0.008 (Cd), 7.06 ± 1.58, 3.90 ± 0.86 (Cr), 0.85 ± 0.043, and 0.18 ± 0.02, and 1.12 ± 0.21 (As) mg kg-1. Pb concentration was lower than the detection limit (ND). The results showed that the liver concentrations of Cu, Zn, Fe, Mn, Co, and Cr were significantly higher than those in the muscle. The association of sex, age, region, and sampling period, with a concentration of these elements, revealed that concentration of zinc in the liver and cobalt in the muscle were significantly higher in the male. Also, significantly higher cobalt and zinc concentrations in muscle were seen in the first 6 months of the year. Age-related differences in muscle concentrations were observed for cobalt. The concentration of trace elements and heavy metals in the liver and muscle samples were not correlated. Comparison of heavy metals concentration in both tissues with European Commission regulation showed that except Cd, the other heavy metals had a higher level than the EU standard. The results of this study showed that camel meat can be contaminated with heavy metals, but more investigations are needed.

Seasonal Variation and Relationships Between Copper of Serum and Various Tissues in Copper Poisoned Sheep in Kerman Province, Iran

Background: Soil and plant contamination in copper exposure causes chronic copper poisoning (CCP) in animals following the consumption of plants in these soils. Objectives: The present study was carried out on 10 clinically affected copper poisoned cases (in each season) from autumn 2015 to the end of summer 2016, in four seasons in Kerman province to evaluate the seasonal effects of CCP in sheep. Methods: All samples were taken from clinically affected cases of copper poisoning that were then necropsied and the poisoning was confirmed by pathognomic pathological findings and serum copper concentration. Samples from liver, lung, kidney, heart and spleen were collected immediately after death and their copper concentration was measured. Results: The highest levels of copper in lung, heart, spleen and kidney were observed in the summer. Liver had the highest amounts of copper in spring and the levels of copper in wool and serum were detected in winter. The copper levels of liver and kidney were positively correlated in autumn, but the copper levels in these tissues were negatively correlated in both spring and winter. Conclusions: Severity of the copper toxicity (judged by the liver copper concentration) is season-dependent in sheep and seasonal variations affect this toxicity. Environmental climate and stressors may be the main causes of copper contents of different tissues and the liver is the main organ to reserve copper in poisoned sheep.

Effects of Copper Sources and Levels on Lipid Profiles, Immune Parameters, Antioxidant Defenses, and Trace Element Residues in Broilers.

The study was conducted to investigate the effects of copper sources and levels on lipid profiles, immune parameters, antioxidant defenses, and trace element contents of meat and liver in Arbor Acres broilers. A total of 504 male broilers were randomly divided into 7 groups with 6 replicates per group and 12 broilers per replicate. The experiment was used in a 3 × 2 + 1 factorial experiment design; broilers in the control group were fed a basal diet, and broilers in the other six groups were fed basal diets supplemented with 3 sources (copper sulfate, tribasic copper chloride, and copper methionate) and 2 levels (10 and 20mg/kg). The results showed that the levels of cholesterol and low-density lipoprotein cholesterol in broilers were significantly decreased with the increase of dietary copper level (P <0.05). Serum IL-6 and IgA contents, ceruloplasmin and GSH-Px activities, and liver copper contents of broilers increased significantly with dietary copper levels (P <0.05). Compared with the control group, dietary copper supplementation significantly decreased serum cholesterol (P <0.05) and significantly increased serum IL-6, ceruloplasmin, SOD, GSH-Px, and liver copper (P <0.05). Dietary supplementation of basic copper chloride and copper methionate significantly decreased low-density lipoprotein cholesterol content and liver iron content (P < 0.05). In conclusion, dietary copper supplementation can effectively reduce serum cholesterol content and improve immune and antioxidant functions in broilers. Adding 20mg/kg copper to broiler diet can increase the copper content in the liver, but it will not affect the copper content in the chicken.

Assessment of corneal and lens clarity in children with Wilson disease

To investigate the effect of copper accumulation on corneal and lens clarity in children with Wilson disease (WD) compared to healthy children. This multicenter cross-sectional study included 24 subjects with WD and 25 age-matched controls. Clinical and laboratory characteristics of the WD subjects were recorded. The Pentacam HR imaging system was used both for lens densitometry and corneal densitometry. Corneal densitometry values were higher in the posterior 6-10mm(P=0.021), posterior 10-12mm(P<0.001), posterior total diameter (P=0.037), total thickness 10-12mm(P=0.032), and total thickness 6-10mm zones and layers (P=0.040) in the WD eyes than in control eyes. The lens densitometry values of zone 1 were higher in WD eyes (P<0.001). There was a significant relationship between corneal densitometry values in the posterior 10-12mm zones (P=0.012; r=0.527) and the duration of WD and liver copper content (P=0.016; r=0.507). A statistically significant correlation was also detected between lens densitometry values in zone 1 and WD duration (P=0.018; r=0.426). In this study cohort, children with WD had decreased corneal and lens clarity even in cases without Kayser-Fleischer rings and sunflower cataracts. Densitometry measurements using Scheimpflug imaging provided detection of corneal and lens involvement in the early stages of WD.

Unique Aspects of Copper Binding Components in the Blood Plasma of Dogs

It is well known that dogs suffer from at least two unusual traits related to Cu metabolism, which are (a) an inability of their serum albumin to bind Cu tightly, due to no histidine in the N terminal binding site; and (b) 10–20‐times higher liver Cu concentrations accompanied by a tendency to develop Cu overload.[1,2] We have identified and investigated two additional peculiarities in dogs related to components in their blood plasma. One has to do with ceruloplasmin (Cp), a 132 kDa glycoprotein, which has functions ranging from delivery of Cu to cells and scavenging reactive oxygen species, to oxidation of Fe(II) and various amines. It is also the main Cu binding component of the blood plasma. Although liver Cu concentrations are extremely high compared to other species, total Cu levels in plasma are normally much lower than those in human and rats (200–400 ng/ml versus about 1200). We found that when the plasma of Labrador retrievers was fractionated in large pore size exclusion chromatography (SEC), the main Cu peak eluted much earlier than is the case with plasma from other mammals. In Superdex 200 FPLC equilibrated with 20 mM phosphate, pH 7, the main Cu peak eluted near the void volume (Mr ~ 700 kDa) as compared to that of humans mice, rats and pigs (Mr~150 kDa). The same was the case for plasma from other dog breeds. Western blotting determined that Cp protein eluted in parallel with the main Cu peak and was of normal size in SDS‐PAGE. Enzyme activities associated with Cp (ferroxidase and p‐phenylenediamine oxidase) also eluted in parallel. This indicated that canine Cp was either bound to some other protein(s) or that it was aggregating. Lactoferrin, which has been reported to bind to Cp in milk, was not detected. Canine plasma proteins were further fractionated on Sephacryl S300, which gives a better separation of large components. Stained SDS‐PAGE gels of fractions from such columns did not show any bands that eluted in parallel with those of Cp. This is consistent with the concept that Cp was not binding another protein but aggregating. To further examine that possibility, fractionation was carried out in buffers of higher ionic strength. Isotonic phosphate‐buffered saline did not alter the elution of canine Cp in Superdex 200. However with 300 mM phosphate (pH 6.8), canine Cp eluted in the same way as the Cp of other mammals, indicating it had dissociated. The other canine peculiarity investigated relates to the tendency towards Cu toxicosis. Plasma from Labrador retrievers with mutations in the Cu transporter ATP7B and high liver copper concentrations had markedly increased levels of a small Cu carrier in their plasma and urine compared with the wild type, as is the case with Wilson disease model mice.[3] Our results indicate that increased levels of a small Cu carrier may be helpful in diagnosis of canine Cu overload, and suggest that canine Cp is circulating as a multimeric protein.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Short communication: Survey of hepatic copper concentrations in Midwest dairy cows

Previous research from our laboratory and others indicates that liver copper concentrations in dairy cattle are commonly well above those recognized as adequate for the nutritional needs of the animal. It has also been speculated that hepatic copper concentrations have been increasing in recent years. Unlike other species, the threshold at which elevated liver copper concentrations becomes deleterious to hepatocytes is not known for cattle. Therefore, the objectives of this study were 3-fold: (1) to delineate differences in the range and mean dry matter hepatic copper concentration for dairy cattle in a retrospective analysis (January 1, 2007, to December 31, 2015), (2) to investigate hepatic copper concentrations in Midwest cull dairy cattle, and (3) to evaluate histologic changes in hepatocellular morphology in the context of copper concentration in cull cows. Furthermore, microscopic changes in hepatocellular morphology or architecture were examined and scored for evidence of inflammation, fibrosis, necrosis, and abundance of rhodanine-stained granules using hematoxylin and eosin and rhodanine staining. The retrospective analysis found copper concentrations within a range of 3 to 1,963 µg/g, with a mean of 473 µg/g. Hepatic copper concentrations in our retrospective study did not increase with time. In our abattoir analysis, copper concentrations ranged from 15 to 978 μg/g, with a mean of 390 μg/g. This study found that the range and mean hepatic copper concentrations were comparatively less in the current abattoir study than copper concentrations in our retrospective analysis. There was no evidence for hepatocellular changes associated with increased copper burdens in this study population.

Age and diet-specific trace element accumulation patterns in different tissues of chub (Squalius cephalus): Juveniles are useful bioindicators of recent pollution

Chub (Squalius cephalus L. 1758) specimens of three age groups with different types of diet were collected in November 2013 in the River Szamos/Someş, Hungary. The Ca, K, Mg, Na, Cd, Cr, Cu, Fe, Mn, Pb, Sr and Zn concentrations were analyzed in the muscle, gills and liver samples of chub by microwave assisted plasma-atomic emission spectrometry (MP-AES). The Kruskal–Wallis test revealed significant differences among different age groups based on the trace element concentrations in the liver, muscle and gills. The trace element concentration pattern in muscle and liver of different age groups differed, may be according to the different diet types of the groups. Meanwhile no differentiation among the age groups based on the trace element concentration in the gills was observed, probably because the pattern of trace elements in the gills is related to the habitat preference, which does not differ during the life-span of chub. In contrast to expectations, trace element concentrations in juveniles were the highest in most cases, certainly because of their specific diet, relatively fast metabolic rate and inadequately developed detoxification system. Only the copper concentrations in liver increased with fish age. Considerable concentrations of trace elements in the tissues of juveniles were observed in the case of elements whose concentrations in the River Szamos were higher in 2013 than in previous years. According to this phenomenon, trace element patterns in the tissues of juveniles may be good indicators of recent pollution of watercourses.

In Atp7b−/− Mice Modeling Wilson’s Disease Liver Repopulation With Bone Marrow-Derived Myofibroblasts or Inflammatory Cells and Not Hepatocytes Is Deleterious

In Wilson's disease, Atp7b mutations impair copper excretion with liver or brain damage. Healthy transplanted hepatocytes repopulate the liver, excrete copper, and reverse hepatic damage in animal models of Wilson's disease. In Fah-/- mice with tyrosinemia and α-1 antitrypsin mutant mice, liver disease is resolved by expansions of healthy hepatocytes derived from transplanted healthy bone marrow stem cells. This potential of stem cells has not been defined for Wilson's disease. In diseased Atp7b-/- mice, we reconstituted bone marrow with donor cells expressing green fluorescent protein reporter from healthy transgenic mice. Mature hepatocytes originating from donor bone marrow were identified by immunostaining for green fluorescence protein and bile canalicular marker, dipeptidylpeptidase-4. Mesenchymal and inflammatory cell markers were used for other cells from donor bone marrow cells. Gene expression, liver tests, and tissues were analyzed for outcomes in Atp7b-/- mice. After bone marrow transplantation in Atp7b-/- mice, donor-derived hepatocytes containing bile canaliculi appeared within weeks. Despite this maturity, donor-derived hepatocytes neither divided nor expanded. The liver of Atp7b-/- mice was not repopulated by donor-derived hepatocytes: Atp7b mRNA remained undetectable; liver tests, copper content, and fibrosis actually worsened. Restriction of proliferation in hepatocytes accompanied oxidative DNA damage. By contrast, donor-derived mesenchymal and inflammatory cells extensively proliferated. These contributed to fibrogenesis through greater expression of inflammatory cytokines. In Wilson's disease, donor bone marrow-derived cells underwent different fates: hepatocytes failed to proliferate; inflammatory cells proliferated to worsen disease outcomes. This will help guide stem cell therapies for conditions with proinflammatory or profibrogenic microenvironments.

Serum and liver copper, iron, molybdenum and zinc concentration in goats and sheep in the state of Paraíba, Brazil

ABSTRACT: This study aimed to determine Cu, Fe, Mo, and Zn liver and serum concentration in sheep and goats raised in the rangelands of the semiarid region of the state of Paraíba, Brazil, during the dry and rainy seasons, and to establish if Cu deficiency is primary or secondary to high ingestion of Mo or Fe. Cu, Zn, Mo and Fe concentrations were determined by atomic absorption spectrometry coupled to mass (ICP-Plasma) in 253 liver and serum samples randomly selected in a slaughterhouse. The mean serum concentrations of Cu in the goats and sheep were 11.82±3.28μmol/L and 10.97±3.61μmol/L respectively. The liver Cu concentrations were 160.37±11.77mg/kg in goats and 152.12±13.16mg/kg in sheep. The mean serum Fe concentrations were 16.38±4.51μmol/L in goats and 25.41±9.76μmol/L in sheep. The mean Fe concentrations in the liver were 189.37±6.51mg/kg in goats and 313.70±12.89mg/kg in sheep. The mean serum concentrations of Mo were 0.14±0.04μmol/L in goats and 0.29±0.06μmol/L in sheep. The mean Mo concentrations in the liver were 6.09±0.23mg/kg in goats and 6.22±0.15mg/kg in sheep. The mean serum Zn concentrations were 8.30±1.91μmol/L in goats and 8.63±2.22μmol/L in sheep. The mean Zn concentrations in the liver were 132.80±3.39mg/kg in goats and 130.70±2.99mg/kg in sheep. These results show low or marginal serum and liver concentrations of Cu and Zn, indicating that these minerals should be supplemented. The normal or even low concentrations of Mo and the high concentration of Fe suggest that Cu deficiency may be primary, due to low Cu ingestion, or secondary due to high Fe ingestion.

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.

Copper Requirements of Pregnant Ewes under Conditions of the Arid Zone in Russia

The copper requirements of pregnant ewes in the environment of the arid zone in the Republic of Kalmykia have been studied. The organ and tissue analyses have shown that the copper quantitative concentrations in the fetal tissues and organs tend to vary depending on its age. The maximum copper contents are recorded in the bone tissue and the skin (1839.7 and 1714.08 μg%, respectively) of the fetuses of 100 days of age and in the bone tissue and the liver (1843.08 and 1261.04 μg%, respectively) of the 145-day-old fetuses. In the bone tissue, the percentage of the element was relatively steady during the period of the prenatal development and varied in the range of 1839.7–1843.08 μg%. The blood copper level has slightly increased for the survey period; the blood mass increased 5.84-fold. The total amount of copper in the blood of the fetus at the age of 100 days was 111.21 μg%, while it increased up to 717.33 μg% in the blood of the 145-day-old fetus. For the period of the fetal development, the copper contents in the muscle tissue and the sheepskin with wool decreased from 957.06 to 52.53 μg% and 1714.08 to 170.7 μg%, respectively, while the total amount of this element in these tissues decreased from 3531.07 to 1020.5 μg and from 1458.65 to 1309.05 μg, respectively. The liver copper content elevated with the gestational age by 37.14 μg%, it comprised 1261.04 μg% up to the age of 145 days, and the total content increased 3.1 times. The relative percentage and the total content of copper in the lungs increased by 77.29 μg% and 4.3 times, respectively, between 100 and 145 days of age generally due to the increase in the organ masses. The total copper content in the 100-day-old and 145-day-old fetuses comprised 6.95 and 9.74 mg, respectively. The copper doses at the stages of early (55 days), middle (100 days), and late (145 days) gestation of the Kalmyk fat-rumped meat sheep were determined to be 12.93, 14.02, and 15.34 mg, respectively. Taking into consideration the fact that the dietary copper absorbtion rate is in the range of 71.05–73%, the fat-rumped ewes entering early, middle, and late pregnancy should intake 18.2, 19.3, and 21 mg dietary copper, respectively. The dietary copper intake norms per 1 kg dry matter intake are 11.0, 7.9, and 8.0 mg; the copper intake norms per 1 kg live weight should be 0.30, 0.32, and 0.35 mg.

Effects of Dietary Copper on Growth Performance, Slaughter Performance and Nutrient Content of Fecal in Growing Goslings from 28 to 70 Days of Age

ABSTRACT A dose-response experiment with four dietary copper concentrations (4.17, 8.17, 12.17 and 16.17 mg/kg) was conducted to estimate the growth performance, slaughter performance, nutrient content of fecal and liver copper concentrations of growing Goslings from 28 to 70 d of age. Two hundred healthy male Yangzhou geese with similar body weight were randomized to four groups with five replicates per treatment and ten geese per replicate. Average daily feed intake, average daily gain and feed conversion ratio of geese for each pen were measured from 28 to 70 d of age. At 70 d of age, two geese were selected randomly from each pen and slaughtered to evaluate carcass quality. Metabolism experiment was conducted with five male geese from each group (one goose per pen) which body weight was close to the mean weight of the group from 64 to 70 d of age. Significant effects of dietary copper was found on body weight, feed conversion ratio, carcass yield, fecal copper concentrations and liver copper concentrations. Body weight, feed conversion ratio and carcass yield showed significant quadratic response to increase dietary copper concentration, while fecal copper concentration and liver copper concentration showed a significant linear response. The result showed that dietary Cu addition can improve growth by increasing the use of the feeding stuff and improving carcass yield in growing Goslings. Furthermore, taking into consideration, the optimal level of Gosling dietary copper was between 8.77 and 11.6 mg/kg from 28 to 70 days of age.