Determination of copper status by five biomarkers in serum of healthy women.

BACKGROUND: The essential trace element copper is relevant for many important physiological processes. Changes in copper homeostasis can result from disease and affect human health. A reliable assessment of copper status by suitable biomarkers may enable fast detection of subtle changes in copper metabolism. To this end, additional biomarkers besides serum copper and ceruloplasmin (CP) concentrations are required. OBJECTIVES: The aim of this study was to investigate the emerging copper biomarkers CP oxidase (CPO) activity, exchangeable copper (CuEXC) and labile copper in serum of healthy women and compare them with the conventional biomarkers total serum copper and CP. METHOD AND MAIN FINDINGS: This observational study determined CPO activity, the non CP-bound copper species CuEXC and labile copper, total serum copper and CP in sera of 110 healthy women. Samples were collected at four time points over a period of 24 weeks. The concentrations of total serum copper and CP were within the reference ranges. The comparison of all five biomarkers provided insight into their relationship, the intra- and inter-individual variability as well as the age dependence. The correlation and Principal Component Analyses (PCA) indicated that CP, CPO activity and total copper correlated well, followed by CuEXC, while the labile copper pool was unrelated to the other parameters. CONCLUSIONS: This study suggests that the non-CP-bound copper species represent copper pools that are differently regulated from total copper or CP-bound copper, making them interesting complementary biomarkers to enable a more complete assessment of body copper status with potential relevance for clinical application.

Direct Measurement of ATP7B Peptides Is Highly Effective in the Diagnosis of Wilson Disease.

BACKGROUND & AIMS: Both existing clinical criteria and genetic testing have significant limitations for the diagnosis of Wilson disease (WD), often creating ambiguities in patient identification and leading to delayed diagnosis and ineffective management. ATP7B protein concentration, indicated by direct measurement of surrogate peptides from patient dried blood spot samples, could provide primary evidence of WD. ATP7B concentrations were measured in patient samples from diverse backgrounds, diagnostic potential is determined, and results are compared with biochemical and genetic results from individual patients. METHODS: Two hundred and sixty-four samples from biorepositories at 3 international and 2 domestic academic centers and 150 normal controls were obtained after Institutional Review Board approval. Genetically or clinically confirmed WD patients with a Leipzig score >3 and obligate heterozygote (carriers) from affected family members were included. ATP7B peptide measurements were made by immunoaffinity enrichment mass spectrometry. RESULTS: Two ATP7B peptides were used to measure ATP7B protein concentration. Receiver operating characteristics curve analysis generates an area under the curve of 0.98. ATP7B peptide analysis of the sequence ATP7B 887 was found to have a sensitivity of 91.2%, specificity of 98.1%, positive predictive value of 98.0%, and a negative predictive value of 91.5%. In patients with normal ceruloplasmin concentrations (>20 mg/dL), 14 of 16 (87.5%) were ATP7B-deficient. In patients without clear genetic results, 94% were ATP7B-deficient. CONCLUSIONS: Quantification of ATP7B peptide effectively identified WD patients in 92.1% of presented cases and reduced ambiguities resulting from ceruloplasmin and genetic analysis. Clarity is brought to patients with ambiguous genetic results, significantly aiding in noninvasive diagnosis. A proposed diagnostic score and algorithm incorporating ATP7B peptide concentrations can be rapidly diagnostic and supplemental to current Leipzig scoring systems.

Selenium and Copper as Biomarkers for Pulmonary Arterial Hypertension in Systemic Sclerosis.

Circulating selenoprotein P (SELENOP) constitutes an established biomarker of Se status. SELENOP concentrations are reduced in inflammation and severe disease. Recently, elevated SELENOP levels have been suggested as diagnostic marker and therapeutic target in pulmonary arterial hypertension (PAH). We decided to re-evaluate this hypothesis. A group of healthy controls (n = 30) was compared with patients suffering from systemic sclerosis (SSc, n = 66), one third with SSc-related PAH. Serum was analysed for trace elements and protein biomarkers, namely SELENOP, glutathione peroxidase 3 (GPx3) and ceruloplasmin (CP). Compared to controls, patients with SSc-related PAH displayed reduced serum Se (91 ± 2 vs. 68 ± 2 µg/L) and SELENOP concentrations (3.7 ± 0.8 vs. 2.7 ± 0.9 mg/L), along with lower GPx3 activity (278 ± 40 vs. 231 ± 54 U/L). All three biomarkers of Se status were particularly low in patients with skin involvement. Serum Cu was not different between the groups, but patients with SSc-related PAH showed elevated ratios of Cu/Se and CP/SELENOP as compared to controls. Our data indicate that patients with SSc-related PAH are characterized by reduced Se status in combination with elevated CP, in line with other inflammatory diseases. Further analyses are needed to verify the diagnostic value of these TE-related biomarkers in PAH.

Deferasirox Might Be Effective for Microcytic Anemia and Neurological Symptoms Associated with Aceruloplasminemia: A Case Report and Review of the Literature.

The patient was a 64-year-old man presented with difficulty in walking, articulation, and swallowing, as well as cognitive impairment. He had refractory microcytic anemia and diabetes mellitus. His serum levels of iron, copper, and ceruloplasmin were low. Magnetic resonance imaging suggested iron deposition in the basal ganglia, thalami, cerebellar dentate nuclei, and cerebral and cerebellar cortices. He was diagnosed with aceruloplasminemia after a ceruloplasmin gene analysis. Iron chelation therapy with deferasirox improved his anemia and cerebellar symptoms, which included dysarthria and limb ataxia. The present study and previous reports indicate that cerebellar symptoms with aceruloplasminemia might respond to deferasirox in less than one year.

Effect of anti-inflammatory compounds or antibiotic administration on receiving performance and physiological responses of transported heifers.

The objectives of this experiment was to evaluate the effects of products with anti-inflammatory properties (yeast product [YEA; 20 g/heifer daily] or astragalus polysaccharide [APS; 20 g/heifer daily]) or an antibiotic (TUL, tulathromycin; 0.025 mL/kg body weight [BW]) on receiving performance and stress responses of transported heifers. Angus heifers (n = 80) were ranked by BW (315 ± 6 kg) and assigned to one of four treatments (five pens per treatment, four heifers per pen) 7 d before shipping 1,400 km (day -7): 1) fed a basal diet of ad libitum hay and concentrate supplement (CON) from day -7 to day 29; 2) YEA in supplemental concentrate from day -7 to day 7 (YEA); 3) APS in supplemental concentrate from day -7 to day 7 (APS); 4) administration of TUL at loading for shipping (day 0; TUL). Upon arrival at the receiving facility (day 1), heifers within each treatment were ranked by BW and assigned to 20 feedlot pens in the same manner as pre-transport. Daily dry matter intake (DMI) was recorded from day 1 to day 28. Full BW was recorded on days -7, -1, 0, 1, 28, and 29. Blood samples were collected on days -7, -1, 1, 4, 7, 14, and 28. Over the receiving period, average daily gain (ADG) and gain: feed did not differ (P ≥ 0.19) for YEA, APS, and TUL, which were greater (P ≤ 0.01) than CON. Average daily gain was also lower (P < 0.01) for CON vs. YEA, APS, and TUL from day -7 to day 28. During the first week of receiving, hay, concentrate, and total DMI were lower (P < 0.01) in CON than the YEA, APS, and TUL, but did not differ (P ≥ 0.13) among these three groups. Hay and total DMI were still lower (P < 0.01) in CON vs. TUL in the second week. Total DMI was greater (P = 0.01) for TUL vs. YEA, and greater (P < 0.01) for YEA vs. CON. Serum nonesterified fatty acid concentrations were greater (P ≤ 0.05) for CON and TUL vs. YEA and APS on day 1. Plasma cortisol concentrations were greater (P ≤ 0.05) for YEA and CON vs. APS and TUL on day 1. Serum tumor necrosis factor-α concentrations were lower (P ≤ 0.05) for APS vs. CON, YEA, and TUL on days 1 and 4. Plasma haptoglobin concentrations were greater (P ≤ 0.05) for CON vs. YEA, APS, and TUL on days 1 and 4, greater (P ≤ 0.05) for YEA, APS vs. TUL on day 1, and greater (P = 0.03) for YEA vs. TUL on day 4. Plasma ceruloplasmin concentrations were greater (P ≤ 0.05) for CON vs. YEA, APS and TUL vs. APS on days 1, 4, and 7. In conclusion, YEA, APS, and TUL modulated the physiological stress responses and alleviated the performance losses caused by long-distance transportation.

[Severe hypocupremia and familial amyloid polyneuropathy]. / Hipocupremia severa y polineuropatía amiloidótica familiar por transtiretina.

INTRODUCTION: Introduction: we report a patient with transthyretin familial amyloid polyneuropathy (TTR-FAP) and severe hypocupremia. Case report: a 79-year-old male with TTR-FAP and severe malnutrition. Laboratory tests showed low serum copper (Cu) and ceruloplasmin levels, as well as low urinary Cu levels. The patient reported neither digestive symptoms nor previous gastrointestinal surgery. Liver function tests, iron metabolism, hemoglobin, leukocytes and zinc were normal. Discussion: Cu is a trace element. It is part of the cuproenzymes involved in several physiological functions. Hypocupremia can be related to genetic or acquired etiologies, including low intake, bariatric surgery, increased losses, etc. Primary clinical manifestations include hematological (anemia and leukopenia) and neurological (myelopathy, peripheral neuropathy) features. Treatment is empirical. In severe cases it may be initiated with endovenose administration, followed by oral supplementation.

INTRODUCCIÓN: Introducción: presentamos el caso de un paciente con antecedentes de polineuropatía amiloidótica familiar por transtiretina (TTR-FAP) diagnosticado de hipocupremia severa. Caso clínico: varón de 79 años afecto de TTR-FAP. Visto en consulta de nutrición por desnutrición severa. En el estudio analítico presenta cifras de cobre (Cu) sérico y ceruloplasmina bajas, con Cu en orina también bajo. No tiene clínica digestiva ni antecedentes de cirugía gastrointestinal. Las pruebas de función hepática, la ferrocinética, las cifras de Hb y leucocitos y los niveles de zinc (Zn) no presentan alteraciones relevantes. Discusión: el Cu es un oligoelemento que participa como componente de las cuproenzimas en múltiples funciones fisiológicas. Los niveles séricos bajos pueden relacionarse con causas genéticas o adquiridas, como la baja ingesta, la cirugía bariátrica, el aumento de las pérdidas, etc. Las principales manifestaciones clínicas son hematológicas (anemia, leucopenia) o neurológicas (mielopatía, neuropatía periférica). El tratamiento tiene base empírica. En los casos severos puede iniciarse con administración intravenosa, seguido de mantenimiento por vía oral.

Characterization of Fibrinogen as a Key Modulator in Patients with Wilson's Diseases with Functional Proteomic Tools.

Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism caused by defects in the ATPase gene (ATP7B). The various clinical features result from the massive accumulation of copper in the liver, cornea and basal ganglia. Although WD can be effectively treated with proper medicine, this disease is difficult to clearly diagnose due to its indefinite symptoms. In the current study, we achieved a positive correlation between clinical symptoms and the enzymatic activity of ceruloplasmin in WD patients. Furthermore, proteome profiles of plasma as well as network analysis demonstrated that fibrinogen is a critical indicator which is significantly unregulated in WD subjects in comparison to healthy donors and closely linked to pathogenesis of WD. Here, we applied 2DE-immunoblots and immunohistochemistry to verify the protein level and localization in situ. The enhanced expression of fibrinogen in the plasma of WD subjects with respect to that of healthy controls and patients with distinct disorders was also confirmed by utilizing clinical samples. As expected, application of high dose of copper induced expression of fibrinogen, while knockdown of ceruloplasmin also resulted in upregulation of fibrinogen as well as elimination of superoxide dismutase (SOD), leading to increased oxidative stress in cells. In summary, the liver injury or oxidative stress induced by the progression of WD may account for the obvious increase of fibrinogen, which in turn triggers inflammatory responses and interferes coagulation cascades; this finding sheds light on the early detection and diagnosis of WD.

The interaction between resveratrol and two forms of copper as carbonate and nanoparticles on antioxidant mechanisms and vascular function in Wistar rats.

BACKGROUND: Experimental studies have emphasized that cardiovascular alterations can be improved by the long-term use of resveratrol (trans-3,5,4'-trihydroxystilbene; RSV) as well as dietary copper (Cu) intake. METHODS: Male Wistar rats were supplemented for 8 weeks with Cu (6.5 mg/kg diet) as either nanoparticles (40 nm, CuNPs) or carbonate (CuCO3). Half of the studied animals were supplemented with RSV (500 mg/kg diet). Vascular function and blood plasma antioxidant status, expressed as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), lipid hydroperoxides (LOOH) and malondialdehyde (MDA) were analyzed. The activity of ceruloplasmin (Cp), lipid profile, fasting glucose, and concentrations of Cu and zinc (Zn) were analyzed. RESULTS: RSV supplementation resulted in the elevated activity of SOD and decreased CAT, GPx and LDL-cholesterol in both groups. RSV supplementation on CuNPs increased the participation of vasoconstrictor prostanoids and decreased ACh-induced vasodilation, while the participation of hyperpolarizing mechanism(s) was restored by activating KATP channels. Blood plasma glucose was decreased. RSV supplementation on CuCO3 enhanced ACh- and SNP-induced vasodilation and decreased NA-induced vasoconstriction. The lipid profile was improved, as well as Zn concentration. Meanwhile, Cu and Cp, and the markers of lipid peroxidation, reflected as LOOH and MDA, were decreased. CONCLUSION: The use of RSV during CuCO3 intake improves vascular responses, the lipid profile and the antioxidant mechanism(s). The beneficial role of RSV was not observed in the CuNP group and decreased ACh-induced vasodilation and increased participation of vasoconstrictor prostanoids in the vascular regulation were noticed.

A STROBE compliant observational study on trace elements in patients with ulcerative colitis and their relationship with disease activity.

Nutritional deficiencies and malnutrition are considered to be related to ulcerative colitis (UC); however, the association between serum levels of micronutrients and UC is not well known. This study aimed to evaluate the serum levels of micronutrients in UC patients and investigate their association with disease activity.This cross-sectional study was conducted on UC patients visiting the Department of Gastroenterology at 3 different teaching hospitals between January 2016 and January 2017. UC activity was measured based on Truelove and Witts' severity index and guidelines for colonoscopy. A healthy gender- and age-matched group was also selected. Serum levels of zinc, copper, selenium, ceruloplasmin, albumin, and total protein were compared between the 2 groups of UC patients and healthy subjects using independent-samples t test. Also, the association between serum levels of micronutrients and UC activity was assessed by using Pearson and Spearman correlation coefficient tests. The data were analyzed by SPSS version 21, considering P ≤.05 as the statistical significance level.Overall, 112 (54 male and 58 female) individuals with the mean age of 34.6 years were studied in the 2 groups of UC patients (n = 56) and healthy subjects (n = 56). The 2 groups were homogeneous in terms of age, gender, marital status, place of residence, and educational level (P >.05). The serum levels of total protein (6.41 ±â€Š1.1 vs 7.41 ±â€Š0.4 g/dL; P = .0001), albumin (4.72 ±â€Š1.1 vs 5.19 ±â€Š0.28 g/dL; P = .0001), zinc (679 ±â€Š62 vs 1055 ±â€Š156 µg/L; P = .0001), and selenium (81.85 ±â€Š6.4 vs 108.4 ±â€Š12.98 micg/L; P = .0001) were significantly lower in the UC patients. The serum level of copper did not differ significantly between the 2 groups (P = .1).Considering the simultaneous reduction in nutritional criteria in the UC patient group, malnutrition appears to be a factor affecting micronutrient deficiency in patients with UC.

Parâmetros hematológicos e bioquímicos de bezerros neonatos da raça Holandesa tratados com ferro suplementar / Hematological and biochemical parameters of neonatal Holstein calves supplemented with iron

A deficiência de ferro em bezerros neonatos está associada ao desenvolvimento de anemia, que favorece o aparecimento de outras enfermidades como pneumonia e diarreia. Avaliou-se o efeito da suplementação de ferro sobre o eritrograma, teores séricos de ferro, ceruloplasmina e transferrina, bem como o potencial para toxicidade do protocolo utilizado por meio da avaliação dos teores de ureia, creatinina e enzimas hepáticas. Para tal avaliação foram utilizados 40 bezerros neonatos da raça Holandesa, alocados em cinco grupos experimentais com oito animais em cada grupo, que foram submetidos aos seguintes protocolos: administração intramuscular de 5mL de solução fisiológica estéril no 5º dia de idade (grupo controle G1), e administração intramuscular de 5mL de ferro dextrano 10% nos seguintes momentos: no 5º dia de idade (G2); no 5o e no 20º dias de idade (G3); no 5o e no 30º dias de idade (G4) e no 5o, 20o e 45º dias de idade (G5). Foram coletadas amostras de sangue até 8 horas após o nascimento e aos 5, 10, 20, 30, 60 e 90 dias de idade para realização do eritrograma, avaliação dos teores séricos de ferro, ceruloplasmina, transferrina, ureia, creatinina, bilirrubina total e direta, e das atividades das enzimas aspartato aminotransferase (AST), fosfatase alcalina (ALP) e gamaglutamiltransferase (GGT). Os animais que receberam ferro suplementar apresentaram menor oscilação nos parâmetros eritrocitários, embora os animais do grupo controle não tenham desenvolvido anemia. Notou-se também aumento, embora não significativo, nos teores séricos de ferro e das proteínas de fase aguda ceruloplasmina e transferrina, cuja atividade está relacionada ao metabolismo desse mineral. Os teores séricos de ureia, creatinina, bilirrubina total e direta e as atividades das enzimas GGT, AST e ALP não foram influenciados pelos protocolos de administração de ferro suplementar. Os protocolos de tratamento empregados não ocasionaram hepatoxidade ou nefrotoxidade aos animais. Concluiu-se que a suplementação com ferro dextrano por via parenteral em bezerros que recebem outras dietas que não apenas leite não traz benefícios que justifiquem sua indicação, embora sejam necessários mais estudos que avaliem a influência da suplementação com ferro sobre o tempo necessário para a recuperação, custos com o tratamento e impacto sobre a vida produtiva dos animais na idade adulta.(AU)

Iron deficiency in newborn calves is associated with the development of anemia, which favors the development of other infirmities such as pneumonia and diarrhea. The present study evaluated the effect of iron supplementation on erythrogram, serum levels of iron, ceruloplasmin and tranferrin, as well as potential toxicity of the protocol used by means of evaluation of urea, creatinine and hepatic enzyme activities. 40 newborn Holstein calves were allocated into 5 experimental groups comprising 8 calves each, which were subjected to the following treatment protocols: intramuscular administration of 5mL of sterile saline on the 5th day of age (control group G1), intramuscular administration of 5mL of 10% dextran iron in the following moments: on the 5th day of age (G2); on the 5th and in the 20th day of age (G3); on the 5th and 30th day of age (G4); on the 5th, 20th and 45th days of age (G5). Blood samples were taken until 8 hours after birth and with 5, 10, 20, 30, 60, and 90 days of age, and subjected to hemogram, evaluation of serum levels of iron, ceruloplasmin, transferrin, urea, creatinine, total and direct bilirrubin, and serum activities of aspartate aminotransferase (AST), alcaline phosphatase (ALP), and gamma-glutamyltransferase (GGT). Calves that received iron supplementation at any time presented less variation in the erythrocyte parameters, although calves in the control group did not develop anemia. Serum concentration of iron and acute phase protein ceruloplasmin and transferrin, which activities are related to iron metabolism, also increased, although not significantly. Serum levels of urea, creatinine, bilirubins and activities of AST, ALP, and GGT were not influenced by the administration protocols used in this experiment. The results of the experiment led to the conclusion that the supplementation with parenteral dextran iron in calves that receive diets other than exclusive milk does not bring sufficient advantages to be indicated, although more studies are necessary to evaluate the influence of iron supplementation on the outcome of infections in newborn calves, especially its influence on cost of treatment, time necessary for discharge and impact on its productive life.(AU)

Zinc containing dental fixative causing copper deficiency myelopathy.

A 62-year-old male, previously well, was referred to neurology clinic following 6 months history of worsening lower limbs instability, paraesthesia, pain and weakness rendering him housebound. Examination revealed upper motor neuron pattern of weakness of the lower limbs and loss of proprioception. Serum analysis revealed reduced caeruloplasmin and copper levels with raised zinc. Spinal imaging revealed subtle dorsal column intensity changes in C2-C7, confirmed with 3T MRI. A copper deficiency myeloneuropathy was diagnosed secondary to chronic use of a zinc-containing dental fixative paste. The paste was discontinued and a copper supplementation was started. Resolution of symptoms was not achieved with intensive physiotherapy. The patient remains a wheelchair user though progression of symptoms has halted. Prompt recognition and treatment of hyperzincaemia-induced hypocupraemia earlier in the disease course may have prevented any irreversible neurological deficit.

MEK and ERK Kinase Inhibitors Increase Circulating Ceruloplasmin and Cause Green Serum in Rats.

Inhibition of the mitogen-activated protein kinase/extracellular signal-regulated (MAPK/ERK) pathway is an attractive therapeutic approach for human cancer therapy. In the course of evaluating structurally distinct small molecule inhibitors that target mitogen-activated protein kinase kinase (MEK) and ERK kinases in this pathway, we observed an unusual, dose-related increase in the incidence of green serum in preclinical safety studies in rats. Having ruled out changes in bilirubin metabolism, we demonstrated a 2- to 3-fold increase in serum ceruloplasmin levels, likely accounting for the observed green color. This was not associated with an increase in α-2-macroglobulin, the major acute phase protein in rats, indicating that ceruloplasmin levels increased independently of an inflammatory response. Elevated serum ceruloplasmin was also not correlated with changes in total hepatic copper, adverse clinical signs, or pathology findings indicative of copper toxicity, therefore discounting copper overload as the etiology. Both ERK and MEK inhibitors led to increased ceruloplasmin secretion in rat primary hepatocyte cultures in vitro, and this increase was associated with activation of the Forkhead box, class O1 (FOXO1) transcription factor. In conclusion, increased serum ceruloplasmin induced by MEK and ERK inhibition is due to increased synthesis by hepatocytes from FOXO1 activation and results in the nonadverse development of green serum in rats.

Influencing the Tumor Microenvironment: A Phase II Study of Copper Depletion Using Tetrathiomolybdate in Patients with Breast Cancer at High Risk for Recurrence and in Preclinical Models of Lung Metastases.

PURPOSE: Bone marrow-derived progenitor cells, including VEGFR2+ endothelial progenitor cells (EPCs) and copper-dependent pathways, model the tumor microenvironment. We hypothesized that copper depletion using tetrathiomolybdate would reduce EPCs in high risk for patients with breast cancer who have relapsed. We investigated the effect of tetrathiomolybdate on the tumor microenvironment in preclinical models. EXPERIMENTAL DESIGN: Patients with stage II triple-negative breast cancer (TNBC), stage III and stage IV without any evidence of disease (NED), received oral tetrathiomolybdate to maintain ceruloplasmin (Cp) between 8 and 17 mg/dL for 2 years or until relapse. Endpoints were effect on EPCs and other biomarkers, safety, event-free (EFS), and overall survival (OS). For laboratory studies, MDA-LM2-luciferase cells were implanted into CB17-SCID mice and treated with tetrathiomolybdate or water. Tumor progression was quantified by bioluminescence imaging (BLI), copper depletion status by Cp oxidase levels, lysyl oxidase (LOX) activity by ELISA, and collagen deposition. RESULTS: Seventy-five patients enrolled; 51 patients completed 2 years (1,396 cycles). Most common grade 3/4 toxicity was neutropenia (3.7%). Lower Cp levels correlated with reduced EPCs (P = 0.002) and LOXL-2 (P < 0.001). Two-year EFS for patients with stage II-III and stage IV NED was 91% and 67%, respectively. For patients with TNBC, EFS was 90% (adjuvant patients) and 69% (stage IV NED patients) at a median follow-up of 6.3 years, respectively. In preclinical models, tetrathiomolybdate decreased metastases to lungs (P = 0.04), LOX activity (P = 0.03), and collagen crosslinking (P = 0.012). CONCLUSIONS: Tetrathiomolybdate is safe, well tolerated, and affects copper-dependent components of the tumor microenvironment. Biomarker-driven clinical trials in high risk for patients with recurrent breast cancer are warranted. Clin Cancer Res; 23(3); 666-76. ©2016 AACR.

Inclusion of anti-phospholipase A2 antibody to backgrounding diets on performance, feed efficiency, in vitro fermentation, and the acute-phase response of growing beef calves.

In Exp. 1, individual performance and daily DMI was measured on 70 crossbred weaned calves during a 70-d period using a GrowSafe system (GrowSafe Systems Ltd., Airdrie, AB, Canada) at the University of Florida North Florida Research and Education Center Feed Efficiency Facility (FEF). Calves were fed a low-concentrate (LC) growing diet, blocked by weight and sex, and then randomly assigned to pens to receive either no additional supplement (CON; n = 35) or receive a supplement of anti-phospholipase A2 antibody (aPLA2) at an inclusion rate of 0.6% of the diet DM (n = 35). After the 70-d feed efficiency (FE) trial (Phase 1), calves were loaded into a commercial livestock trailer and were driven for approximately 1,600 km during 24 h. Upon return to the FEF (Phase 2), calves were relocated to the same pens and groups and received the same diets and treatments for 28 d. Blood samples from each calf were collected on d 0, 1, 3, 5, 7, 14, 21, and 28 relative to initiation of transportation and were analyzed for determination of concentrations of plasma ceruloplasmin and haptoglobin. In Phase 1, initial BW (242.0 ± 3.7 kg; P = 0.92), BW at d 70 (313.0 ± 4.1 kg; P = 0.79), and ADG (1.01 ± 0.02 kg; P = 0.95) were similar between treatments. However, daily DMI was greater (P = 0.01) for CON (9.18 ± 0.15 kg) than aPLA2 (8.53 ± 0.15 kg). In addition, residual feed intake was greater (P = 0.002) for CON (0.389 ± 0.110 kg/d) than aPLA2 calves (-0.272 ± 0.110 kg/d). In Phase 2, after transportation, there were no differences between treatments on BW loss due to transportation shrink (26.0 ± 0.6 kg; P = 0.86), BW at d 28 (339.0 ± 4.1 kg; P = 0.72), ADG (1.28 ± 0.03 kg/d; P = 0.72), G:F (0.164 ± 0.004; P = 0.83), and concentrations of plasma haptoglobin (0.08 ± 0.02 mg/mL; P = 0.41). However, concentrations of plasma ceruloplasmin were greater (P < 0.001) for CON calves (14.3 ± 0.3 mg/dL) compared to aPLA2 calves (13.0 ± 0.3 mg/dL). In Exp. 2, the effects of aPLA2 inclusion on LC and high-concentrate (HC) substrates on in vitro fermentation parameters were assessed. Addition of aPLA2 had no effects on in vitro fermentation parameters of LC and HC substrates. In conclusion, supplementation of aPLA2 improved FE of growing beef calves when fed LC diets in Phase 1 and addition of aPLA2 had no effect on fermentation parameters of LC and HC substrates. In addition, calves supplemented with aPLA2 had reduced concentrations of plasma ceruloplasmin after 24 h of transportation.

Liver transplantation in neurological Wilson's Disease: is there indication? A case report.

Wilson's disease (WD) is an autosomal recessive disorder characterized by copper overload. In this disease, inadequate hepatic excretion leads to copper accumulation in the liver, brain, kidney, and cornea. Severe neurological symptoms can develop in patients with WD, often in the absence of relevant liver damage: it is unclear whether liver transplantation (LT) could reverse neurological symptoms, and at present LT is not recommended in this setting. We report a case of regression of neurological symptoms in a patient affected by WD with prevalent neurological involvement. A 19-year-old man with disabling neuropsychiatric symptoms from WD that included frontal ataxia, akinesia, dystonia, tremors, and behavioral disorders in the presence of preserved liver function (Model for End-Stage Liver Disease score=7; Child-Turcotte-Pugh score=A5) underwent LT in November 2009. At the time of LT, encephalic magnetic resonance imaging (MRI) indicated diffuse neurodegenerative alterations involving subtentorial and supratentorial structures; bilateral Kayser-Fleischer ring was present. Four years after LT, laboratory tests show normalized copper metabolism and excellent liver function test results. Encephalic MRI shows a substantial improvement of already-known signal alterations at nuclei thalamus and putamen, mesencephalon, and pons. Kayser-Fleischer ring disappeared from the right eye, but a little remnant is still visible in the left eye. At neurological examination, all of the previous symptoms and signs are no longer present and behavioral disorders are no longer present; psychosocial functions are completely restored. The present case provides some evidence that LT may be a valid therapeutic option for WD patients with marked neurological impairment, particularly in those no longer responsive to chelation therapy.

Effect of pomegranate juice supplementation on matrix metalloproteinases 2 and 9 following exhaustive exercise in young healthy males.

OBJECTIVES: To evaluate the efficacy of pomegranate juice supplementation on matrix metalloproteinases2 and 9 serum levels and improving antioxidant function in young healthy males during exhaustive exercise. METHODS: The study was conducted at Ardabil University of Medical Sciences, Iran, in 2010-11 and comprised 28 healthy subjects in 18-24 age bracket. They were randomly divided into control and supplemented groups. One cup of pomegranate juice and one cup of tap water were given to supplemented and control groups daily for two weeks respectively. Fasting blood samples were taken at baseline and at the end of two weeks of intervention. The subjects were given one exhaustive exercise and then fasting blood samples were taken for testing blood glutathione peroxidase and superoxide dismutase and serum levels of high sensitivity C-reactive protein, zinc, ceruloplasmin, matrix metalloproteinases 2 and 9, malondialdehyde and total antioxidant capacity. Data was analysed using descriptive statistical tests, paired and independent sample t-test. RESULTS: The blood levels of glutathione peroxidase and superoxide dismutase and serum levels of total antioxidant capacity after exhaustive exercise in the supplemented group were significantly increased (p < 0.05), while the content of matrix metalloproteinases 2 and 9, ceruloplasmin and malondialdehyde showed a significant decrease in comparison to the control group (p < 0.05). Besides, there were no significant changes in other biochemical factors. CONCLUSION: Regular intake of pomegranate juice significantly modulates matrix metalloproteinases 2 and 9and serum levels of some inflammatory factors and thus protects against exhaustive exercise-induced oxidative injury in young healthy males.

Anemia and leukopenia in a long-term parenteral nutrition patient during a shortage of parenteral trace element products in the United States.

Recently, drug shortages in the United States have affected multiple components of the parenteral nutrition (PN) solution. A 62-year-old patient with systemic sclerosis who was dependent on home PN due to intestinal dysmotility developed anemia and leukopenia approximately 4 months after parenteral copper was withheld from her PN solution due to drug shortages. The patient was not able to tolerate a sufficient amount of oral multivitamins with trace elements due to severe dysphagia. Her serum copper and ceruloplasmin concentrations were undetectable, confirming the diagnosis of severe copper deficiency. The hematological abnormalities promptly resolved with copper supplementation. This report emphasizes the importance of close monitoring for nutrient deficiencies during drug shortages and supplementing with oral or enteral nutrition when feasible, particularly in high-risk patients such as those with intestinal malabsorption or short bowel syndrome who are dependent on long-term PN.

Iron supplementation in pregnancy and breastfeeding and iron, copper and zinc status of lactating women from a human milk bank.

This study evaluated the influence of iron supplementation in pregnancy and breastfeeding on iron status of lactating women from a Brazilian Human Milk Bank. Blood and mature breast milk samples were collected from 145 women for assessment of iron status, as well as copper and zinc status. Haemoglobin, serum iron and ferritin were determined, respectively, by electronic counting, colorimetry and chemiluminescence. Transferrin and ceruloplasmin were analysed by nephelometry. Serum copper and zinc were measured by atomic absorption spectrophotometry, and serum alkaline phosphatase was measured by a colorimetric method. Iron, zinc and copper in breast milk were determined by spectrometry. Mean values of iron, copper and zinc (blood and breast milk) were compared by ANOVA, followed by Tukey's test. Iron supplementation was beneficial to prevent anaemia in pregnancy but not effective to treat anaemia. During breastfeeding, iron supplementation had a negative effect on maternal copper status, confirming an interaction between these micronutrients.