Fenótipo distinto da doença hepática gordurosa não alcoólica em pacientes com níveis reduzidos de cobre livre e ceruloplasmina / Distinct phenotype of non-alcoholic fatty liver disease in patients with low levels of free copper and of ceruloplasmin

ABSTRACT BACKGROUND: Copper deficiency has been linked to alterations in lipid metabolism and hepatic steatosis. Oxidative stress plays a role in the pathogenesis of non-alcoholic fatty liver disease (NAFLD). One of the enzymes that neutralize oxidative stress is Cu/Zn superoxide dismutase, which depends on the availability of adequate amounts of copper. OBJECTIVE: Correlate the levels of ceruloplasmin and of non-ceruloplasmin-bound copper (NCBC) with clinical, biochemical and histological parameters of non-alcoholic fatty liver disease (NAFLD) patients. METHODS: Data from 95 consecutively admitted NAFLD patients who underwent liver biopsy composed the groups based on ceruloplasmin levels lower than 25 mg/dL and on negative NCBC. The risk factors for NAFLD in each group were compared. RESULTS: Body mass index was lower in patients with ceruloplasmin <25 mg/dL (29.1±3.47 vs 32.8±6.24 kg/m2; P=0.005) as were the levels of LDL, HDL and total cholesterol, when compared with their counterparts with ceruloplasmin >25 mg/dL (101±38 vs 116±35 mg/dL, P=0.05; 43±9 vs 51±16 mg/dL, P=0.01; 174±43 vs 197±39 mg/dL, P=0.01, respectively). Mean serum ferritin levels were higher in the ceruloplasmin <25 mg/dL group (343±327 vs 197±190 ng/mL; P=0.02). Otherwise, patients with negative NCBC had higher HOMA-IR (8.2±14.7 vs 4.6±3.7; P=0.03). Age, gender, hypertension and diabetes showed no statistical difference. CONCLUSION: Patients with NAFLD had different clinical and biochemical markers according to the levels of NCBC and ceruloplasmin.

RESUMO CONTEXTO: A deficiência de cobre tem sido relacionada a alterações no metabolismo lipídico e esteatose hepática. O estresse oxidativo desempenha um papel fundamental na fisiopatologia da doença hepática gordurosa não alcoólica. Uma das enzimas que neutralizam o estresse oxidativo é a Cobre/Zinco superoxido dismutase, que depende da disponibilidade de quantidades adequadas de cobre. OBJETIVO: Correlacionar os níveis de ceruloplasmina e de cobre não ligado à ceruloplasmina (NCBC) com parâmetros clínicos, bioquímicos e histológicos de pacientes com doença hepática gordurosa não alcoólica (DHGNA). MÉTODOS: Dados de 95 pacientes com DHGNA internados consecutivamente e submetidos à biópsia hepática compuseram os grupos com base em níveis de ceruloplasmina inferiores a 25 mg/dL e em NCBC negativo. Os fatores de risco para DHGNA em cada grupo foram comparados. RESULTADOS: O índice de massa corporal foi menor nos pacientes com ceruloplasmina <25 mg/dL (29,1±3,47 vs 32,8±6,24 kg/m2; P=0,005), assim como os níveis de LDL, HDL e colesterol total, quando comparados aos seus pares com ceruloplasmina >25 mg/dL (101±38 vs 116±35 mg/dL, P=0,05; 43±9 vs 51±16 mg/dL, P=0,01; 174±43 vs 197±39 mg/dL, P=0,01, respectivamente). Os níveis médios de ferritina sérica foram maiores no grupo ceruloplasmina <25 mg/dL (343±327 vs 197±190 mg/mL; P=0,02). Os pacientes com NCBC negativo apresentaram maior HOMA-IR (8,2±14,7 vs 4,6±3,7; P=0,03). Idade, sexo, hipertensão e diabetes não mostraram diferença estatística. CONCLUSÃO: Pacientes com DHGNA apresentaram diferentes marcadores clínicos e bioquímicos de acordo com os níveis de NCBC e ceruloplasmina.

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)

Effects of thymoquinone and curcumin on the regeneration of rat livers subject to 70% hepatectomy

Abstract Purpose: To investigate thymoquinone, curcumin and a combination of these two drugs were effective or not at the growth of liver. Methods: Forty female Wistar-Albino rats distributed into five groups of eight rats each, control, thymoquinone, curcumin, and thymoquinone/curcumin groups. Pathological specimens were studied using the Ki-67 Proliferation Index(PI); and arginase(Arg), tissue plasminogen activator(tPA), ceruloplasmin(Cer) and nitric oxide(NO) were studied in biochemical analysis. Results: Our results showed that Ki-67 proliferation index was low in Groups 1. The proliferation coefficient was significantly higher in the Group 2 and Group 4 than in the Group 1 and Group 3.(P < 0.001 between Groups 1 and 2, 1 and 4, and 3 and 4). There was no difference between Groups 2 and 4 (P = 1). The results of the biochemical Arg, tPA and Cer test showed statistically between the Group 1 and Group 2. NO showed significant differences Group 1 and 3. Conclusions: Thymoquinone and curcumin both have known positive effects on the organism. Histological and biochemical tests showed that thymoquinone is more effective than curcumin.

Serum concentrations of acute phase proteins in goats and ewes with naturally acquired Staphylococcus aureus mastitis / Concentração de proteínas de fase aguda em cabras e ovelhas com mastite de ocorrência natural causada por Staphylococcus aureus

Serum protein concentrations, including acute phase proteins (APPs), of goats and ewes with naturally acquired Sthaphylococcus aureus mastitis were determined by means of SDS-PAGE electrophoresis to evaluate the relevance of these APPs as biomarkers of the disease in these species. Fifteen healthy goats and 5 goats with naturally acquired staphylococci mastitis, as well as fifteen healthy ewes and 5 ewes with staphylococci mastitis were submitted to daily blood sampling during 7 days. In goats, an increase of 570%, 125%, 621%, and 279% in serum concentrations of ceruloplasmin, fibrinogen, haptoglobin and α1-acid glycoprotein, respectively, was observed. In sheep the increase in serum concentrations of ceruloplasmin, fibrinogen, haptoglobin and α1-acid glycoprotein was of 337%, 90%, 461%, and 225%, respectively. Our results indicate that these APPs have considerable potencial as early and sensible biomarkers of mastitis caused by S. aureus in goats and sheep.(AU)

O proteinograma, incluindo proteínas de fase aguda (PFAs), de cabras e ovelhas com mastite de origem natural causada por Staphylococcus aureus, foi determinado por meio de eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio (SDS-PAGE) a fim de avaliar a importância destas PFAs como biomarcadores da enfermidade nestas espécies. Amostras de sangue foram colhidas diariamente de cinco cabras e cinco ovelhas com mastite estafilocócica naturalmente adquirida, bem como de quinze cabras e quinzes ovelhas saudáveis durante 6 dias consecutivos. Nas fêmeas caprinas, foi verificado aumento dos teores séricos de ceruloplasmina (570%), fibrinogênio (125%), haptoglobina (621%), e α1-glicoproteína ácida (279%). Nas fêmeas ovinas as concentrações de ceruloplasmina, fibrinogênio, haptoglobina e α1-glicoproteína ácida elevaram-se em 337%, 90,9%, 461% e 225%, respectivamente. Os resultados permitem inferir que estas PFAs são marcadores sensíveis e precoces de mastite causada por S. aureus em cabras e ovelhas.(AU)

Menkes disease: importance of diagnosis with molecular analysis in the neonatal period / Doença de Menkes: importância do diagnóstico com análise molecular no período neonatal

Summary Menkes disease is a congenital disorder caused by changes in copper metabolism derived from mutations in the ATP7A gene. It is characterized by physical and neurological alterations. In the neonatal period, these alterations can be nonspecific, which makes early diagnosis a challenge. Diagnosis can be suspected when there are low levels of ceruloplasmin and serum copper. Molecular analysis confirms the diagnosis. Treatment is parenteral administration of copper histidine. We report a familial case with molecular confirmation. The proband had clinical and biochemical suspicious. Treatment with copper histidine was indicated, but initiated at the age of 2 months and 27 days only. He did not present improvements and died at 6 months. The mother became pregnant again, a male fetus was identified and copper histidine was manufactured during pregnancy. He was born healthy, biochemical markers were reduced and treatment was indicated. Molecular analysis was performed confirming mutation in both the mother and the proband, while the other son did not have mutation, so treatment was discontinued. We support the clinical relevance of molecular confirmation for the correct diagnosis and genetic counseling, once clinical findings in the neonatal period are nonspecific and early treatment with parenteral copper histidine must be indicated.

Resumo A doença de Menkes é causada por uma alteração genética no metabolismo do cobre, por mutações no gene ATP7A. Caracteriza-se por alterações neurológicas e no exame físico. No período neonatal, essas alterações podem ser inespecíficas, o que torna o diagnóstico precoce um desafio. O diagnóstico pode ser suspeitado quando há baixos níveis séricos de cobre e ceruloplasmina. A análise molecular confirma o diagnóstico, e o tratamento deve ser feito com histidina de cobre. Nós relatamos um caso familial de doença de Menkes. O probando apresentava quadro clínico e alterações bioquímicas compatíveis com a doença de Menkes, em consulta com 1 mês de vida. O tratamento foi indicado, mas apenas iniciado com 2 meses e 27 dias. Ele não apresentou melhora clínica e veio a óbito com 6 meses. A mãe teve uma nova gestação, foi identificado um feto do sexo masculino e foi solicitada a manipulação da histidina de cobre ainda durante a gestação. O bebê nasceu saudável, os marcadores bioquímicos estavam diminuídos e o tratamento com histidina de cobre foi indicado. Realizamos a análise molecular, que confirmou mutação no gene ATP7A na mãe e no probando; porém, o outro filho não apresentava mutação e o tratamento foi interrompido. Nós defendemos a importância clínica da confirmação molecular para o correto diagnóstico e o aconselhamento genético da doença de Menkes, uma vez que os achados clínicos e as alterações bioquímicas no período neonatal são inespecíficos, e o tratamento com histidina de cobre parenteral deve ser rapidamente instituído.

Wilson's disease: laboratory evaluation of a sample of patients attending Baghdad teaching hospital

Wilson's disease [WD] is an inherited defect in copper metabolism that causes accumulation of copper in various body organs. It is a treatable; if it is diagnosed promptly and treated consistently. Laboratory; hematological, biochemical, and diagnostic evaluation of a sample of Iraqi patients with WD, and study any association between clinical presentations and studied variables. A case series study was conducted during 2011, from the 1[st] of February till the 10[th] of June. Sampling method was a convenient non-random one, through consecutive pooling of registered WD patients. A questionnaire-form paper had been developed for data collection, and required investigations were done in qualified laboratories. The study had enrolled 29 patients, with a male to female ratio of [1.07:1], The mean hemoglobin level was 10.64 +/- 2.53g/dl. Hemolysis was presented in 27.6% of patients, normochromic-normocytic RBCs in blood film were seen in 3.4% patient, platelet count was 243.97 +/- 89.35ccX10[9]/L, WBC count was 5.70 +/- 2.84ccX10[9]/L. ESR was 24.98 +/- 17.16mm/hr., serum bilirubin was 6.17 +/- 5.18mg/dl, serum cholesterol was 146.21 +/- 44.93mg/dl, serum triglycerides was 95.34 +/- 52.23mg/dl. Total serum protein was 72.86 +/- 11.15g/L, serum Albumin 39.17 +/- 7.88g/L and serum Globulin 31.07 +/- 7.89g/L. Penicillamine challenge test was positive in 96.6% patients. Serum ceruloplasmin level was 123.38 +/- 48.48mg/L, total serum copper was 72.48 +/- 25.11 micro g/dl, and 24h urinary copper excretion was 174.97 +/- 109.58 micro g/24hr. Most patients with WD are; anemic, with low serum levels of copper and cerulloplasmin, positive penicillamine challenge test, with abnormal liver function tests. With detected hemolysis in some of them; though no statistical difference or association was approved regarding laboratory values among patients with different presentations

Analysis of clinical and biochemical spectrum of Wilson Disease patients.

Background and Aims: Wilson disease (WD) is autosomal recessive disorder of copper metabolism. Wilson disease patients usually suffer from hepatic or neuropsychiatric complications. The symptoms appear between ages five to 35 but it can vary from two years to 72 years. Materials and Methods : Study was carried out from June 2008 to November 2010. This study included nine families with eleven cases of WD to determine clinical presentation, diagnostic findings (including laboratory results) and liver histology. It included 11 patients who presented with hepatic manifestations and/or Neuropsychiatric manifestations and/or family history suggesting features of WD. Patients with hepatitis B and C and those with history of taking antipsychotic drugs were excluded from the study. Patient's data was included in a well designed performa. Liver function test, serum ceruloplasmin, serum copper, 24 hour urinary copper, blood complete picture were analyzed. Quantitative data such as age, hemoglobin etc were expressed as mean with ± SD and quantitative variables such as sex, movement disorders, hepatic involvement etc were expressed as frequency and percentage. Results: There were five male and six female patients with evidence of various manifestations here (i) hepatic in which they had only liver dysfunction (ii) hepatic and neurological (iii) neurological. The mean age of presentation was 8.7±3.92 years (range 4-19 years) and 45% were male patients. Decreased serum ceruloplasmin, enhanced 24-h urinary copper excretion and signs of chronic liver damage were confirmed in all patients and Kayser-Fleischer rings (KF rings) in 72% of patients. In severe WD patients, serum prothrombin activity was less than 50%, serum ceruloplasmin were low and serum copper levels were high than those in non-severe WD patients. High degree of suspicion leads to early treatment with good outcome. Conclusions: The WD is rare but important cause of chronic liver disease. Clinical and biochemical analysis in cases of patients with unexplained liver disease with high degree of suspicion can lead to early treatment with good outcome.

Effects of Urtica dioica on hepatic ischemia-reperfusion injury in rats

OBJECTIVES: To evaluate the effects of Urtica dioica on hepatic ischemia-reperfusion injury. METHODS: Thirty adult male Wistar albino rats were divided into three groups: sham group (group 1), control group (group 2), and Urtica dioica group (group 3). All the rats were exposed to hepatic ischemia for 60 min, followed by 60 min of reperfusion. In group 2, a total of 2 ml/kg 0.9 percent saline solution was given intraperitoneally. In group 3, a total of 2 ml/kg Urtica dioica was given intraperitoneally. At the end of the procedure, liver tissue and blood samples were taken from all rats. Serum aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, ceruloplasmin, catalase, paraoxonase, arylesterase, and lipid hydroperoxide levels were measured. Liver tissue histopathologies were also evaluated by light microscopy. RESULTS: Serum aspartate aminotransferase, alanine aminotransferase and lactate dehydrogenase levels were significantly higher in group 2 than in group 1, and significantly lower in group 3 than in group 2. Also, group 2 had higher serum lipid hydroperoxides and ceruloplasmin levels but lower catalase, paraoxonase, and arylesterase levels than group 1. In group 3, serum lipid hydroperoxides and ceruloplasmin levels were significantly lower, and catalase, paraoxonase, and arylesterase levels were higher than those in group 2. Histopathological examination showed that liver tissue damage was significantly decreased in group 3 compared with group 2. CONCLUSIONS: Urtica dioica has a protective effect on the liver in hepatic ischemia-reperfusion-injured rats.

Doença de Menkes: relato de caso / Menkes' Disease: case report

Relata-se um caso clássico de doença de Menkes, uma doença neurodegenerativa acompanhada de manifestações sistêmicas, dentre elas aspecto típico do cabelo. O diagnóstico se confirma pelos níveis baixos de ceruloplasmina e cobre no sangue. Os autores trazem uma revisão atualizada, não sistemática, da literatura.

A classical case of Menkes? disease is discussed, a neurodegenerative disease accompanied by systemic manifestations, including a typicalaspect of the hair. The diagnosis is confirmed by low serum levels of ceruloplasmin and copper. The authors bring forth an updated nonsystematic revision of the literature.

Serum micronutrient and micromineral concentrations and ratios in healthy Omani subjects

This study aimed to establish reference ranges of serum concentrations of copper, zinc, retinol, alpha-tocopherol, copper: caeruloplasmin and copper: zinc ratios in a group of healthy Omani men and women. Assay techniques employed were atomic absorption spectrophotometry [copper and zinc], reverse-phase high-pressure liquid chromatography with isocratic elution [retinol and alpha-tocopherol], immunonephelometry [caeruloplasmin] and spectrophotometry [albumin and cholesterol]. The mean +/- SD [microM] obtained for copper, zinc, retinol, and alpha-tocopherol were 15.9 +/- 3.0, 14.2 +/- 2.0, 1.45 +/- 0.39 and 16.9 +/- 4.4, respectively. The mean +/- SD for copper: zinc and copper: caeruloplasmin ratios were 1.15 +/- 0.30 micromol/mmol and 6.99 +/- 0.84 micromol/g, respectively. Significantly higher [p < 0.0001] copper and caeruloplasmin concentrations, copper: zinc and copper: caeruloplasmin ratios and lower zinc, retinol, alpha-tocopherol, cholesterol concentrations and alpha-tocopherol: cholesterol ratio were present in women compared to men. Age appeared to be associated with copper and retinol concentrations, and copper: caeruloplasmin ratios in women; in men, the association was mostly moderate with caeruloplasmin, alpha-tocopherol, cholesterol concentrations and alpha-tocopherol: cholesterol ratios. Smokers had decreased albumin [p = 0.002], zinc [p = 0.023] concentrations, and copper: caeruloplasmin ratios [p = 0.002], increased alpha-tocopherol concentrations [p = 0.016] and alpha-tocopherol: cholesterol ratios [p = 0.021] compared with non-smokers. Deficiency incidence was

Ceruloplasmina: determinación de su actividad ferroxidasa / Ceruloplasmin: measurement of serum ferroxidase activity

La Ceruloplasmina (Cp) es la principal proteína transportadora de cobre en circulación. Su concentración es abundante en plasma; se considera un reactante de fase aguda y su función fisiológica no se encuentra fehacientemente establecida. Fundamentalmente se sintetiza en los hepatocitos. También se encuentra en otros tipos celulares como monocitos, astrocitos y células de Sertoli. Su concentración sérica se utiliza en el diagnóstico diferencial de enfermedad de Wilson. La concentración total en plasma se considera igual a la suma de las concentraciones de apo y holo Cp, de manera que la cantidad de esta proteína determinada por un método inmunológico no indica que la enzima se encuentre presente solamente en su forma activa. Entonces, al utilizar esta metodología, se sobreestima la proteína funcionalmente activa. Existen diversos métodos para determinar su actividad. En este trabajo se describe un método automatizado para medir su actividad ferroxidasa que utiliza iones Fe2+ como sustrato. Los valores de referencia de actividad de Cp se diferenciaron estadísticamente entre el grupo de mujeres y el de hombres, siendo de 424-796 UI/L y 397-733 UI/L, respectivamente. Además, se obtuvo una correlación significativa entre la actividad ferroxidasa y la concentración proteica (r=0,7285; p<0,0001).

Ceruloplasmin (Cp) is the principal copper carrier in human plasma. It is an abundant protein that participates in the acute phase reaction to stress, but its physiological function is unknown. Althought Cp is synthesised predominantly in the liver, other cell types express the protein, including monocytes, astrocytes and Sertoli cells. The serum concentration of the copper protein ceruloplasmin has been an important diagnostic indicator of Wilson`s disease. Measurement of the total amount of Cp protein may not reflect Cp enzyme activity in the serum. The immunologic assay may lead to overestimation of the total amount of functional Cp in the serum due to this method's capacity to determine both the functional holo Cp and non-functional apo Cp. Several methods for determining ferroxidase activity have been reported. In this study, a method is described for automated measurement of the activity. In this method, Fe2+ ions are used as the substrate. The range for serum Cp ferroxidase activity in healthy persons was 424-796 UI/L for women, and 397-733 UI/L for men. Significant correlations between serum ferroxidase activity and Cp concentration (r=0,7285; p < 0,0001) were found.

Associação da concentração plasmática de cobre com metaloproteínas cobre-dependentes em atletas de elite / Association between copper plasma concentration and copper-dependent metaloproteins in elite athletes

O cobre é um elemento-traço essencial para a manutenção de vários processos biológicos, tais como metabolismo energético, homeostase de ferro e mecanismos de proteção antioxidante através da atividade da cobre-zinco superóxido dismutase (Cu-Zn SOD), da ceruloplasmina e da metalotioneína. No entanto, o cobre também participa de reações oxidativas que promovem a liberação de radicais livres, podendo prejudicar a integridade e a funcionalidade celular. A atividade física afeta a homeostase do cobre e promove maior utilização de oxigênio, favorecendo a instalação do estresse oxidativo quando mecanismos naturais de proteção antioxidante, incluindo os dependentes de cobre, não atuam adequadamente. Não há relatos na literatura sobre a associação de diferentes níveis de concentração plasmática de cobre com indicadores antioxidantes cobre-dependentes em atletas de elite. O presente estudo objetivou verificar a associação entre diferentes níveis plasmáticos de cobre e metaloproteínas cobre-dependentes, com atividade antioxidante, em atletas de elite. Os indicadores bioquímicos (metalotioneína e Cu-Zn SOD eritrocitárias, ceruloplasmina e cobre plasmáticos) foram avaliados em 50 atletas, homens e adultos, utilizando metodologias já consolidadas. Os resultados mostraram que 32 por cento dos atletas apresentaram níveis de cobre plasmático inferiores a 11µmol/L, 38 por cento entre 11-13µmol/L e 30 por cento > 13µmol/L. As associações encontradas entre cobre plasmático e ceruloplasmina (r = 0,31; p = 0,04) e Cu-Zn SOD (r = 0,32, p = 0,02); metalotioneína eritrocitária e ceruloplasmina (r = 0,40, p = 0,006) e Cu-Zn SOD (0,73, p = 0,001) e entre Cu-Zn SOD e ceruloplasmina (r = 0,37, p < 0,001) demonstraram que a atividade da Cu-Zn SOD e a concentração de metalotioneína eritrocitárias são sensíveis a menor concentração, enquanto que a ceruloplasmina é sensível a elevadas concentrações plasmáticas de cobre, sugerindo que há um equilíbrio homeostático entre...

Copper is a trace element essential in several biological processes, some of them important for physical activity, such as energy metabolism, iron homeostasis and antioxidant protection through the plasma ceruloplasmin, erythrocyte Cu-Zn superoxide dismutase (Cu-Zn SOD) and metallothionein. However, copper also participates in oxidative reactions releasing free radicals, which may adversely affect cell integrity and function. Physical activity is known to affect copper homeostasis and may interfere in the copper antioxidant capacity. Intense physical activity results in higher oxygen consumption, which favors the release of free radicals and may cause irreversible damage to the body when the natural mechanisms of protection, including those copper-dependent, are not properly stimulated. Few studies related exercise with plasma copper level and copper-dependent metalloproteins in elite athletes. The present study aimed at evaluating the association between different levels of plasma copper and copper-dependent metalloproteins in male elite athletes (n = 50). The biochemical indices studied were plasma copper and ceruloplasmin, and erythrocyte Cu-Zn superoxide dismutase and metallothionein by validated methods. The results showed that 32 percent of the athletes had plasma copper levels lower than 11µmol/L, 38 percent between 11-13 µmol/L and 30 percent higher than 13 µmol/L. Plasma copper was associated with plasma ceruloplasmin level (r = 0.31, p = 0.004), and with Cu-Zn SOD (r = -0.32, p = 0.02); metallothionein erythrocyte were associated with Cu-Zn SOD (r = 0.73, p = 0.001) and with ceruloplasmin (r = 0.40, p = 0.006). These results suggest that both plasma and erythrocyte antioxidant capacity favor homeostatic adjustments in agreement with plasma copper levels in elite athletes.

Menkes' disease: case report

Menkes disease is a rare neurodegenerative disorder due to an intracellular defect of a copper transport protein. We describe a 7 months male patient who presented with seizures, hypoactivity and absence of visual contact. The investigation disclosed pilli torti and thrycorrexis nodosa in the hair, low serum levels of both copper and ceruloplasmin, brain magnetic resonance study showed atrophy and white matter hypointensities on T1-weighted images, electroencephalogram reveals moderate background activity disorganization and epileptiform activity, and muscle biopsy with type 2 fiber atrophy. The clinical, laboratorial, genetic, muscle biopsy and neurophysiological findings in MenkesÆ disease are discussed.

A doença de Menkes é uma rara desordem neurodegenerativa causada por defeito intracelular na proteína transportadora do cobre. Descrevemos um paciente de 7 meses, masculino, com crises convulsivas, hipoatividade e ausência de contato visual. A investigação demonstrou pilli torti e thrycorrexis nodosa; níveis séricos baixos de ceruloplasmina e cobre; RNM de crânio com atrofia e redução de sinal da substância branca (imagens em T1); eletroencefalograma com moderada desorganização da atividade de base e atividade irritativa; e biópsia muscular com atrofia de fibras do tipo 2. As características clínicas, laboratoriais, genéticas, biópsia muscular e estudo neurofisiológico na doença de Menkes são discutidas.

Avaliação dos níveis de hiper-homocisteinemia, vitamina E, selênio, cobre, ceruloplasmina e ferritina em pacientes com diagnóstico de acidente vascular cerebral isquêmico / Assesment of homocysteine, vitamin E, selenium, copper, ceruloplasmin and ferritin levels in patients with ischemic stroke diagnosis

OBJETIVO: Avaliar níveis sangüíneos de hiper-homocisteinemia, vitamina E, selênio, cobre, ceruloplasmina e ferritina em pacientes com diagnóstico de acidente vascular cerebral isquêmico (AVCI). CASUíSTICA, MATERIAIS E MÉTODOS: Estudo transversal analítico de 104 pacientes diagnosticados com AVCI e 98 controles. Dosagem de homocisteína e vitamina E feita por cromatografia líquida de alta performance. O selênio foi dosado por tecnologia de espectrometria de massa por plasma indutivamente acoplado; o cobre o foi por colorimetria; a ceruloplasmina, por nefelometria; e a ferritina, por imunoensaio enzimático conjugado à fluorescência (Elfa). Análise estatística feita com testes t de Student e análise de variância. RESULTADOS: Encontrada hiper-homocisteinemia em 43 por cento dos pacientes e 13 por cento dos controles. Os valores obtidos nas dosagens de vitamina E, cobre, ceruloplasmina e ferritina foram significativamente maiores no grupo paciente do que no grupo controle. Quanto ao selênio, a diferença entre os dois grupos não foi significativa. DISCUSSÃO: Níveis de hiper-homocisteinemia foram maiores que os encontrados na literatura, podendo ser conseqüência da genética ou dos hábitos alimentares da população. Os resultados encontrados para vitamina E, selênio, cobre, ceruloplasmina e ferritina coincidem com os de alguns trabalhos publicados, mas o modelo deste estudo não avalia se os níveis de vitamina E e selênio foram alterados pelo AVCI ou por mudanças nos hábitos dos pacientes. A ferritina e a ceruloplasmina podem ser marcadores de AVCI, e não fator causal da sua patogênese. CONCLUSÃO: A hiper-homocisteinemia é mais prevalente nos pacientes diagnosticados com AVCI. Os níveis de vitamina E, cobre, ceruloplasmina e ferritina mais elevados encontrados nos pacientes não podem ser atribuídos à ocorrência de AVCI.

OBJECTIVE: To assess blood levels of hyperhomocysteinaemia, vitamin E, selenium, copper, ceruloplasmin and ferritin in patients with diagnosis of ischemic stroke (IS). METHODS: 104 patients with the diagnosis of IS and 98 healthy controls had blood homocysteine, vitamin E, selenium, copper, ceruloplasmin and ferritin analyzed. Homocysteine and vitamin E were analyzed by high performance liquid chromatography, selenium by inductively coupled plasma - mass spectrometry, copper by colorimetry, ceruloplasmin by nefelometry and ferritin by enzyme linked fluorescent assay. Statistics was performed with Student's t test and analysis of variance. RESULTS: Frequency of hyperhomocysteinaemia was 43 percent for patients and 13 percent for controls. Concentrations of vitamin E, copper, ceruloplasmin and ferritin were significantly higher in patients. Differences in selenium levels were not statistically significant. DISCUSSION: Hyperhomocysteinaemia levels were higher than those found in literature, possibly due to population's genetics or its nutritional habits. As to vitamin E, selenium, copper, ceruloplasmin and ferritin, our results match some published papers but this study's model cannot stablish whether vitamin E and selenium levels were changed by IS or by new habits acquired by the patients. Ferritin and ceruloplasmin can be a marker for IS instead of causative agent of its pathogenesis. CONCLUSION: Hyperhomocysteinaemia is more prevalent in patients with IS diagnosis. Higher levels of vitamin E, copper, ceruloplasmin and ferritin found in patients cannot be imputed to IS.

ATP7B mutations in families in a predominantly Southern Indian cohort of Wilson's disease patients.

OBJECTIVE: To analyze ATP7B mutations in Wilson's disease (WD) patients from the Indian subcontinent and to correlate these with WD phenotype. METHODS: We studied 27 WD patients from 25 unrelated families. Twenty-two families were from three southern Indian states - Tamil Nadu andhra Pradesh and Kerala. We applied conformation- sensitive gel electrophoresis (CSGE) to screen for the mutations in patients and their families. PCR products exhibiting aberrant patterns in CSGE were subjected to direct DNA sequencing. As siblings affected by WD within a family share identical ATP7B genotype, we compared WD phenotype among affected siblings within families. RESULTS: ATP7B mutations were detected in 22 of the 25 probands -13 were homozygotes and 9 were compound heterozygotes. Eleven novel mutations were detected. Only two common mutations were found: G3182A in 4 (16%) and C813A in 3 (12%) probands. 'Hot spots' for ATP7B mutations were exons 18 and 13. Lack of common dominant mutations prevented correlation of individual ATP7B mutations with WD phenotype. Symptomatic WD in a live sibling was not found in any family. In 8 families, a sibling died of presumed WD - in 6 of these, WD phenotype was identical to that in the proband. CONCLUSIONS: We describe the spectrum of ATP7B mutations including 11 novel mutations in Indian WD patients and document lack of a single dominant mutation. Identical WD phenotype among siblings in only 6 of 8 families with >1 child affected by WD suggests that factors other than ATP7B mutations influence WD phenotype.

Pros and cons of immunochemical and enzymatic method in the diagnosis of Wilson's disease.

BACKGROUND: Immunochemical method of measuring Ceruloplasmin (Cp) levels for the diagnosis of Wilson's disease has replaced enzymatic method for the main reason of being more sensitive and quantitative. SETTINGS AND DESIGN: In this study, we compared both the methods for various factors such as sensitivity, specificity and the time consumed in the diagnosis of Wilson's disease. MATERIALS AND METHODS: Serum samples from patients (n=33) with a provisional diagnosis of Wilson's disease were analyzed for Cp levels by enzymatic copper oxidase method and immunochemical method using polyclonal antibodies specific to Cp embedded in agar. STATISTICAL ANALYSIS: Pearson's regression analysis was performed to compare the two methods. RESULTS: The mean Cp obtained by immunochemical method is 5.87 mm +/- 1.17 and by enzymatic method, it is 0.37 (Optical Density) +/- 0.136. Pearson's Regression analysis of the measurements showed a good correlation with an 'r' value of 0.765 between the two methods. CONCLUSION: A good correlation indicated that these two tests are comparable and thus both these methods can be used together for a definitive and better diagnosis of Wilson's disease.

Concentracion sérica de cobre ®libre¼ y ceruloplasmina en niños con desnutricion aguda grave antes de iniciar la terapia nutricional y al momento de recuperar el apetito en turbo, antioquia, colombia / Serum concentrations of free copper and ceruloplasmin in children with severe acute malnutrition before nutritional therapy and after children have recovered the apetite in turbo, antioquia, colombia

Introduction: copper supplementation therapy has been used in children with acute and severe malnutrition. Scientific evidence has shown that malnourished children with edema have free copper in plasma which could produce oxidative stress. Objective: To compared plasma concentrations of free copper between children with acute and severe malnutrition and a control group. Methodology: Cross sectional study where 66 normal and malnourished children were studied. A longitudinal study (before and after type) design was used including 40 children with severe and acute malnutrition; free copper was merasured by high-resolution capillary electrophoresis; ceruloplasmin and PCR by nephelometry. Results: In the cross sectional study children with marasmus had higher free copper serum concentrations than children with oedematous malnutrition, but the difference was not significant. In the control group this metal was not found. Children with oedema showed significant lower ceruloplasmin concentrations than children with marasmus (p=0.00) while the difference in PCR was also no significant for both groups. When the relationship between free copper serum concentrations and the presence of infection was analyzed no significant differences were obtained. However, serum albumin concentration was significantly lower for children with oedematous malnutrition than the marasmic group (p=0.016). After children recovered the appetite in the longitudinal study, serum free copper concentration decreased for both groups and ceruloplasmin concentration increased but no significant differences were observed. Conclusion: Copper supplementation could be considered as nutritional therapy for undernourished children since the beginning of the nutritional treatment, as it is recommended by WHO.

Introducción: La recuperación de los niños con desnutrición aguda grave incluye suplementación con cobre desde el inicio de la terapia; existen evidencias que niños edematosos pueden tener cobre libre en plasma el cual podría generar estrés oxidativo. Objetivo: Comparar concentraciones séricas de cobre libre y ceruloplasmina en un grupo de desnutridos agudos graves y un grupo control Metodología: estudio transversal al ingreso entre niños con y sin desnutrición en una muestra de 66 sujetos y longitudinal de tipo antes y después en 40 niños desnutridos. Se determinó cobre libre por electroforesis capilar de alta resolución; ceruloplasmina y PCR por nefelometría, además de variables clínicas Resultados: En el transversal, los marasmáticos presentaron niveles de cobre libre mayores que los edematosos sin diferencia significativa; en el grupo control este metal no se detectó. En los edematosos la concentración de ceruloplasmina fue significativamente menor con relación a los marasmáticos (p=0.00) y la PCR no presentó diferencia significativa entre ellos. No se encontraron diferencias significativas entre los niveles de cobre libre y presencia o no de infección. La albúmina se encontró más baja en los edematosos con diferencia significativa respecto a los marasmáticos (p=0.016). En el longitudinal, al recuperar el apetito el cobre libre disminuyó y la ceruloplasmina aumentó en los dos grupos sin diferencia significativa Conclusión: el suplemento de cobre como componente de la terapia nutricional puede ser suministrado a los desnutridos desde el inicio como lo propone el protocolo de la OMS.

Alpha-1 antitrypsin deficiency among Indian children with liver disorders.

AIMS: To determine the frequency of alpha-1 antitrypsin (AAT) deficiency in children with chronic liver disease (CLD) and neonatal cholestasis syndrome (NCS). METHODS: All children with NCS (n=23) or CLD (n=35) attending the Pediatric Gastroenterology Clinic between November 2003 and July 2005 were screened for AAT deficiency using phenotyping through isoelectric focusing of plasma. RESULTS: Of the 58 children studied, 57 had normal PiMM phenotype. One child with CLD had the M1E type of normal variant. None of the patients had the abnormal phenotype PiZZ. CONCLUSION: AAT deficiency is infrequent among children with CLD and NCS in our region.