Liver Copper Research Articles

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.

Paroxysmal kinesigenic dystonia mimics focal seizure

Dystonia refers to a syndrome of involuntary sustained or spasmodic muscle contractions involving co-contraction of the agonist and the antagonist. Paroxysmal kinesigenic dyskinesia is one of the rare episodic hyperkinetic movement disorders, characterized by brief attacks of abnormal involuntary movements comprising of dystonia, chorea, athetosis, hemiballismus or a combination, usually precipitated by sudden movements. A 16 years old male previously healthy presented with 1 year history of involuntary movement descried as a sudden stiffness of the left arm, sometimes involving the left side of the body after sudden movement of the trunk or while standing up after prolonged physical rest. The spells usually lasted up to 30 to 60 seconds with frequency 3-4/week and were painless. The patient never had a loss of consciousness during spells. He had a normal physical and neurological examination. Brain MRI, routine EEG, laboratory tests including liver functions testes, serum ceruplasmin and copper and 24 hours urine copper were all normal. Simple partial seizure disorder was suspected. As part of workup the patient was admitted in epilepsy monitoring unite and had a long term video EEG. Recoded spells were consistent with a focal dystonic posturing in the left side of the body precipitated by sudden active movement with a normal EEG background, without any epileptiform discharges . Carbamazepine 200 mg /day was initiated as a treatment of kinesigenic dystonia with good clinical response. Paroxysmal kinesigenic dystonia can be frequently misdiagnosed with other medical conditions and required careful clinical assessment and work up.

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.

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.

Heavy Metals Contamination in the Tissues of Clarias gariepinus (Burchell, 1822) Obtained from Two Earthen Dams (Asa and University of Ilorin Dams) in Kwara State of Nigeria

This study determined the levels of heavy metals contamination in the tissues of 400 Clarias gariepinus obtained from two dams (Asa and University of Ilorin) in Kwara State of Nigeria. C. gariepinus, (both male and female) between the mean weight of 149-154g and total length of between 28-34cm were investigated between the month of September and December, 2018.&amp;nbsp; Copper (Cu), zinc (Zn), cadmium (Cd), chromium (Cr) and lead (Pb) in flesh, heart, kidney and livers of species from the two dams were determined using Atomic Absorption Spectrophotometer (AAS). The results showed that heavy metals contamination in the tissues sampled from Asa and Unilorin dams were in order of Zinc &amp;gt; Copper &amp;gt; Chromium. Cadmium and lead were not detected. Furthermore, C. gariepinus from Asa dam bioaccumulated heavy metals at higher concentrations than samples from Unilorin dam. Body weight, total length and age were found to be predisposing factors to heavy metals concentrations in the body of fish while sexes had no effect.The heavy metals concentration detected in C. gariepinus sampled from the two dams did not exceed the limits set by WHO and FAO suggesting that the fishes wholesome for human consumption.

Pollution induced change of liver of Oreochromis niloticus: metals accumulation and histopathological response

The present study, pointed to assess heavy metal accumulation like Aluminum (Al), Cadmium (Cd), Chrome (Cr), Cobalt(Co), Copper(Cu), Iron(Fe), Lead (Pb), Manganese(Mn), Nickel(Ni), Selenium(Se) and Zinc(Zn) in water, sediments and liver of Oreochromis niloticus, which collected from sewage water in El-kharja, New Valley, Egypt using inductively coupled plasma mass spectrometry (ICP-MS). Also, the histopathological changes of liver of Oreochromis niloticus were reported. These histopathological changes were detected using different staining types as a pollution biomarker. Iron showed the highest accumulation level in water (9.06±0.86 ppm), sediments (175.1±20.8 ppm) and liver (158.17±38.59 ppm), followed by Al> Mn> Ni> Zn> Pb> Cr> Se> Cu> Co> Cd. However, these elements showed Fe> Al> Ni> Mn> Cr> Cu> Zn> Pb> Cd> Se> Co accumulation trend in sediments. In liver these metals showed Fe> Se>Al> Zn>Mn> Cu>Ni> Cr> Pb> Co> Cd trend. Histopathological examination of fish liver showed signs of progressive alterations such as disorganization of architecture of liver cells, hydropic degeneration and vacuolation of hepatocytes. Also, dilation and congestion in blood sinusoids, hypertrophic and increase in number of küpffer cells were chronicled. Sever deposition of hemosiderin pigments were reported while necrosis with pyknotic nucleus and focal histopathological characters were observed side by side with normal cell. Our results concluded that water, sediment and liver accumulation toxicity tests and histopathological changes of liver may be associated, and these approaches may be used together to describe the environmental state and water quality assessment.

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.

Homeostatic changes of some trace elements in geriatric rats in the condition of oxidative stress induced by aluminum and the beneficial role of resveratrol

BackgroundElderly individuals are exposed to trace element imbalances due to the reduced capacity of their organism to utilize minerals in a direct relationship with many circumstances. Objectives and methodsThe aim of this study was to assess the protective role of resveratrol upon the homeostatic changes of some trace elements in geriatric rats in the condition of oxidative stress induced by aluminum exposure. Forty Wistar rats, 18–20 months old, were divided randomly into four groups (n = 10): control (C) - receiving 1 ml of physiologically saline (P.S.) via intraperitoneal (i.p) administration, E1 - 1 ml of P.S. and 1000 ppb aluminum sulphate (AS) in drinking water ad libitum, E2 - 20 mg/kg−1 resveratrol, i.p. and 1000 ppb AS in drinking water, E3 - 20 mg/kg−1 resveratrol i.p. The groups C and E3 received distilled water as drinking water ad libitum. The i.p administrations were once a week for four weeks period. The levels of oxidative stress marker's were analyzed (glutathione, glutathione' peroxidase, superoxide dismutase, and catalase) of the proteins' (total protein, albumin, and hemoglobin) in serum and also the levels of the main trace elements (copper, zinc, iron, selenium, manganese and magnesium) in blood, liver, kidney and spleen. ResultsSignificant decrease (p < 0.05) of total protein (TP), albumin (ALB), catalase (CAT), increase significant (p < 0.05) of glutathione reductase (GSH-r), glutathione peroxidase (GPx) and superoxide dismutase (SOD) in E1 groups, compared with control, E2, and E3 groups was ascertained. There were also observed significant (p < 0.05) decreases in Cu, Zn, Fe and Mg, not significant (p > 0.05) increase of Se and Mn in blood, significant (p < 0.01) increase of Cu, Zn, Mg, Se, Mn in kidney and liver and Fe, in spleen of geriatric rats from E1 group compared to the control group. Insignificant differences (p > 0.05) were recorded in groups which received resveratrol (E2 and E3) compared to the control group, but significant differences (p < 0.05), especially in blood and liver samples, compared to E1. ConclusionsThe results suggest that resveratrol can prevent the homeostatic imbalance of trace elements in geriatric rats in the condition of oxidative stress induced by aluminum exposure.

Copper Deficiency in Liver Diseases: A Case Series and Pathophysiological Considerations.

Copper is an indispensable trace element. It serves as a cofactor for enzymes involved in cellular energy metabolism, antioxidant defense, iron transport, and fibrogenesis. Although these processes are central in the pathogenesis of liver disorders, few studies have attributed them to copper deficiency. We herein describe in detail a case series of liver disease patients (n = 12) who presented with signs of copper deficiency based on serum and liver copper measurements. Median age of the group at the time of presentation was 39 (range 18‐64 years). Six patients were female. The median serum copper was 46 μg/dL (normal range: 80‐155 μg/dL for women and 70‐140 μg/dL for men). Seven of the 12 patients had hepatic copper concentration less than 10 μg/g dry weight (normal range: 10‐35 μg/g). Most cases presented with acute‐on‐chronic liver failure (n = 4) and decompensated cirrhosis (n = 5). Only 3 patients had a condition known to be associated with copper deficiency (ileocolonic Crohn’s disease following resection n = 1, Roux‐en‐Y gastric bypass n = 2) before presenting with hepatic dysfunction. Notable clinical features included steatohepatitis, iron overload, malnutrition, and recurrent infections. In 2 of the 3 patients who received copper supplementation, there was an improvement in serum copper, ceruloplasmin, and liver function parameters. Conclusion: Copper deficiency in the serum or liver occurs in a wide range of liver diseases. Given the biological essentiality of copper, the mechanism and clinical significance of this association require systematic study.

Some heavy metals residues in beef and sheep Liver in Anbar province

The aim of the study To see the effect of location, season and display time on the accumulation of some heavy metals (Lead, copper, zinc, cadmium and cobalt) in sheep and cattle liver. Liver samples were obtained from three districts in Anbar province (Ramadi, Hit and Baghdadi). This study began from January 2017 to November 2017 for all seasons (winter, spring, summer and autumn) Samples were taken from the liver immediately after slaughter for cattle and sheep at 8 am and taken at 4 pm for all areas of study. The results of the study were summarized in the quadratic overlap of the animal type, season, location and time as follows: The highest concentration of lead in liver (38.86 μg / g liver) was recorded in sheep for spring and evening in Ramadi. The lowest concentration was in sheep in Baghdadi for morning and winter (20.06 micro g / g liver). The highest concentration of copper in the liver (34.65 microgram / g liver) was recorded in cows in Ramadi for the winter season and evening time. The lowest concentration was in the sheep liver during the summer season for morning time in the city of al-Baghdadi (20.43 microgram / g liver). The highest concentration was in the liver (603.99 microgram / g liver) in the sheep for the autumn season and the evening time in the city of Ramadi. The lowest concentration was in the sheep in the city of Baghdadi for morning and summer (560.32 microgram / g liver). The highest concentration of cadmium in the liver (30.88 micro g / g liver) was in sheep in Ramadi for the autumn and evening season, and the lowest concentration was in beef in the summer season for morning time in al-Baghdadi city (9.66 μg / g liver). Cobalt was the highest concentration (1.34, 1.34 and 1.35 micro g / g liver) in cow and sheep liver, autumn, summer and evening time for Ramadi. While the lowest concentration of sheep and cattle liver for the city of Baghdadi for the winter season and morning time (0.64 and 0.63 microgram / g liver).

The effect of high levels of dietary zinc on growth performance, carcass characteristics, blood parameters, immune response and tissue minerals in growing Boer-cross goat kids

An experiment was conducted to determine animal performance, carcass traits, blood metabolites, immune responses, and tissue minerals in goats fed high levels of zinc (Zn). Twenty-one Boer-cross (Capra aegagrus hircus) growing goat kids (BW, 22.9 ± 1.4 kg, 4–5 months old) were housed individually and fed twice a day 80:20 grain mix: bermudagrass hay (BGH) diet ad libitum for 86 days. Goats in control group (0Zn) received orally 5 mL of water (1.95 + 0.03 ppm Zinc) with no added Zn. Treatment groups, 100Zn and 200Zn received 5 mL water containing additional 100 or 200 mg Zn as Zn sulfate heptahydrate, respectively. Average daily gain (ADG), DM intake, feed conversion ratio (gain: feed), carcass traits, majority of blood attributes and metabolites, and humoral immunity were similar among the treatment groups. However, cell-mediated immune response was higher for 100Zn group (quadratic; P = 0.04). Goats receiving high levels of Zn increased serum calcium (linear; P = 0.03), glucose (linear, P = 0.02), and lymphocytes (linear, P = 0.05 and quadratic; P = 0.01). Manganese (Mn) level in the liver increased (linear, P = 0.01), while liver copper (Cu) decreased (linear, P = 0.04) as the level of Zn was increased in the diet. Zinc in the feces increased (linear, P = 0.001), with no changes in urine Zn. Blood mineral phosphorus was reduced (linear, P = 0.03), and magnesium, and potassium tended to decrease (linear, P = 0.06 and 0.07, respectively) with increasing Zn levels in diet. High levels of Zn supplementation did not affect animal performance and carcass characteristics; however, improved immunity and impacted liver Mn and Cu.

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.

Characteristics of neurological Wilson\u2019s disease with corpus callosum abnormalities

BackgroundWilson’s disease (WD) is an autosomal recessive disease of impaired copper metabolism. Previous study demonstrated that WD with corpus callosum abnormalities (WD-CCA) was limited to the posterior part (splenium). This study aimed to compare clinical features between WD-CCA and WD without corpus callosum abnormalities (WD-no-CCA).MethodsForty-one WD patients who had markedly neurological dysfunctions were included in this study. We retrospectively reviewed clinical, biochemical characteristics and MRI findings in the 41 WD patients. All patients were assessed using the Unified Wilson’s Disease Rating Scale.ResultsNine patients had corpus callosum abnormalities, 4 of 9 patients had abnormal signal in the genu and splenium, 5 of 9 patients had abnormal signal only in the splenium. WD-CCA had longer course (9.9 ± 4.0 years vs. 3.4 ± 3.6 years, p<0.01), more severe neurological dysfunctions (37.6 vs. 65.9, p<0.01) and higher psychiatric symptoms scores (11.2 vs. 22.5, p<0.01) than WD-no-CCA. The MRI findings indicated that WD-CCA had higher ratio than WD-no-CCA in globus pallidus (88.9% vs. 43.8%, p = 0.024) and thalamus (100% vs. 59.4%, p = 0.038). The index of liver function and copper metabolism had no significant in WD-CCA and WD-no-CCA patients.ConclusionOur findings indicate Wilson’s disease can involve the posterior as well as the anterior part of CC and patients with CC involvement had more extensive brain lesions, more severe neurological dysfunctions and psychiatric symptoms.

Selenium prevents lithium accumulation and does not disturb basic microelement homeostasis in liver and kidney of rats exposed to lithium.

Lithium has been used in medicine for almost seventy years. Besides beneficial effects, its therapy may cause serious side-effects, with kidney and liver being the organs most vulnerable to its harmful influence. Therefore, research on protective agents against lithium toxicity has been continuing for some time. The aim of the present study is to evaluate the influence of additional selenium supplementation on lithium content, as well as homeostasis of the essential microelements iron, zinc, copper and manganese in kidney and liver of rats undergoing lithium exposure. The study was performed on 4 groups of male Wistar rats (6 animals each) treated with: control - saline; Li-group - Li2CO3 at a dose of 2.7 mg Li/kg b.w.; Se-group - Na2SeO3 at a dose of 0.5 mg Se/kg b.w.; Li+Se-group - both Li2CO3 and Na2SeO3 at doses of 2.7 mg Li/kg b.w. and of 0.5 mg Se/kg b.w., respectively, in the form of water solutions by stomach tube, once a day for 3 weeks. The content of the studied elements in the organ samples was determined using flame atomic absorption spectroscopy (FAAS). Lithium administered alone caused a significant increase in its content in liver and kidney. Additional supplementation with selenium reversed these effects, and did not markedly affect other studied microelements compared to control. The obtained results suggest that selenium could be regarded as an adjuvant into lithium therapy. However, considering the limitations of the present study (the short duration, using only one dose and form of selenium) the continuation of the research seems to be necessary to clarify the influence of selenium supplementation on basic microelements and lithium accumulation in organs during lithium exposure.

Levels of trace elements and potential toxic elements in bovine livers: A trend analysis from 2007 to 2018.

Trace elements and potential toxic elements were analyzed in bovine livers submitted for autopsy in the Netherlands during the years 2007 to 2018. The age of each animal was recorded. In total, 1544 livers were analyzed for cadmium, cobalt, chromium, copper, iron, molybdenum, nickel, lead, selenium and zinc. Less than 2% of the liver samples were from veal calves. Young animals had significantly higher concentrations of iron and zinc in their livers compared to animals older than one year, while older animals had significantly higher levels of cadmium and molybdenum in their livers. Animals aged 1 to 2 years had the lowest copper and selenium levels. There was a tendency for lower chromium and nickel levels during the last years of the testing period, while copper showed an increase. Lead intoxication was only seen in the youngest group of cattle, while copper intoxication, defined as a liver copper of more than 1000 mg/kg dry matter, occurred in older animals, mainly in animals of 3 to 4 years old. This trend analysis of trace elements in bovine livers of cattle over time in recent years, and the relation of liver element concentrations with age of the animal, provides insight in the uptake and storage of these elements by cattle in The Netherlands. Possible reasons for observed trends and age-related patterns are discussed.

Pluripotent stem cell-derived bile canaliculi-forming hepatocytes to study genetic liver diseases involving hepatocyte polarity

Hepatocyte polarity is essential for the development of bile canaliculi and for safely transporting bile and waste products from the liver. Functional studies of autologous mutated proteins in the context of the polarized hepatocyte have been challenging because of the lack of appropriate cell models. The aims of this study were to obtain a patient-specific hepatocyte model that recapitulated hepatocyte polarity and to employ this model to study endogenous mutant proteins in liver diseases that involve hepatocyte polarity. Urine cell-derived pluripotent stem cells, taken from a patient with a homozygous mutation in ATP7B and a patient with a heterozygous mutation, were differentiated towards hepatocyte-like cells (hiHeps). HiHeps were also derived from a patient with MEDNIK syndrome. Polarized hiHeps that formed in vivo-like bile canaliculi could be generated from embryonic and patient urine cell-derived pluripotent stem cells. HiHeps recapitulated polarized protein trafficking processes, exemplified by the Cu2+-induced redistribution of the copper transporter protein ATP7B to the bile canalicular domain. We demonstrated that, in contrast to the current dogma, the most frequent yet enigmatic Wilson disease-causing ATP7B-H1069Q mutation per se did not preclude trafficking of ATP7B to the trans-Golgi Network. Instead, it prevented its Cu2+-induced polarized redistribution to the bile canalicular domain, which could not be reversed by pharmacological folding chaperones. Finally, we demonstrate that hiHeps from a patient with MEDNIK syndrome, suffering from liver copper overload of unclear etiology, showed no defect in the Cu2+-induced redistribution of ATP7B to the bile canaliculi. Functional cell polarity can be achieved in patient pluripotent stem cell-derived hiHeps, enabling, for the first time, the study of the endogenous mutant proteins, patient-specific pathogenesis and drug responses for diseases where hepatocyte polarity is a key factor. This study demonstrates that cells that are isolated from urine can be reprogrammed in a dish towards hepatocytes that display architectural characteristics similar to those seen in the intact liver. The application of this methodology to cells from patients diagnosed with inherited copper metabolism-related liver diseases (that is, Wilson disease and MEDNIK syndrome) revealed unexpected and novel insights into patient mutation-specific disease mechanisms and drug responses.

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.