Copper In Liver Tissues Research Articles

Osteoporose em bubalinos em relação à idade, à condição clínica animal e ao estado físico-químico ósseo e do cobre hepático

O presente trabalho faz parte de um programa de pesquisa de doenças metabólicas nutricionais em bubalinos no Trópico Úmido Amazônico e, nesse contexto, objetivou-se estudar os processos osteoporóticos em bubalinos criados em sistema de pastejo na Ilha de Marajó e a relação da enfermidade com a faixa etária e a condição físico-química óssea e do cobre hepático. Foram utilizados 110 animais: 39 jovens e 71 adultos, apresentando Índice de Condição Corporal entre 1 e 2. Avaliou-se um grupo composto por animais jovens e outro por animais adultos. Análises anátomoclínica e da qualidade óssea possibilitaram subdividir cada grupo em dois subgrupos, segundo a presença ou não de manifestações osteopáticas aparentes. Do terço médio da 11ªcostela direita analisou-se o cálcio, fósforo, cinza e densidade óssea e do tecido hepático determinou-se a concentração do cobre. No exame anatomopatológico, respectivamente 98,44% dos animais jovens e 96,16% dos animais adultos apresentavam algum grau de osteoporose (+, ++, +++). Foram demonstrados baixos percentuais na média do fósforo (10,69%), nas cinzas (60,24%) e na densidade (1,46 g/ml). O cobre hepático apresentou valores médios igualmente baixos (19,51 mg/kg). Os resultados mostraram que a osteoporose apresenta um quadro mais severo nos animais jovens, sendo que o processo pode estar relacionado com as baixas reservas de fósforo no tecido ósseo e do cobre no tecido hepático.

Wilson disease—A practical approach to diagnosis, treatment and follow-up

Wilson disease is an inherited, autosomal recessive, copper accumulation and toxicity disorder that affects about 30 individuals per million. This rare disease is caused by mutations in the gene encoding a copper-transporting P-type ATPase, which is important for copper excretion into bile, leading to copper accumulation in the liver. Toxic copper concentrations can also be found in the brain and kidney, and clinical phenotypes include hepatic, haemolytic, neurologic and psychiatric diseases. Diagnosis is based on the combination of clinical features and findings such as increased urinary copper excretion, reduced levels of serum ceruloplasmin, high concentrations of copper in liver tissues and Kayser–Fleischer rings. Genetic studies are also becoming available for clinical use, but the utility of direct mutation analysis is limited. Wilson disease can be treated, and early diagnosis is essential: the goal of therapy is to reduce copper accumulation either by enhancing its urinary excretion or by decreasing its intestinal absorption. Medical therapies include penicillamine, trientine, zinc and tetrathiomolibdate. Liver transplantation is a relatively successful treatment option when medical therapy fails or in case of acute liver failure, even though it is also characterized by short- and long-term complications.

A simple and rapid stain for copper in liver tissue.

We report a modified copper stain that can be completed in 2 hours, rather than overnight, which is required for other stains. This is useful in the diagnosis of chronic cholestatic syndromes, Wilson's disease, Indian childhood cirrhosis, and other conditions associated with copper retention.

肝組織内の銅結合蛋白の組織化学ならびに定量値との比較

A quantitative analysis of copper in liver tissue and a qualitative analysis of copper-binding protein in the histologic sections were subjected to correlative study. Liver sections, obtained from autopsies and surgical resections, were examined histopathologically by orcein staining, rhodanine staining, and other special stainings, in addition to hematoxylin eosin (HE) staining. The quantitative analysis was carried out by an atomic absorption spectrophotometry. Copper and copper-binding protein were not detectable by HE staining. The amount of orcein positive granules (OPGs) was graded semiquantitatively from negative (-) to three plus ( ). The average copper content was 24.9 micrograms/g dry liver weight (micrograms/g) in (-) group, 60.9 micrograms/g in (+) group, 158.9 micrograms/g in (++) group, and 299.3 micrograms/g in ( ) group respectively. Although the copper content and the amount of OPGs were relatively well correlated as mentioned above, standard deviations of copper contents were comparatively large in each group, especially in (++) or ( ) group. Based upon the comparative study of staining qualities of various dyes, rhodanine staining and orcein staining were specific for copper or copper-binding protein. Finally, orcein staining was thought to be the most excellent method from the viewpoints of sensitivity, specificity, contrast and simplicity. The distribution of excessive copper in pathologic conditions was fairly uneven. We recommend a histopathologic examination by using many liver sections stained by orcein, when the condition of copper overload is suspected.

Simple and sensitive determination of hepatic copper by use of an extractive catalytic method

For determination of copper in liver tissue and in liver biopsy samples an extractive catalytic determination is proposed. After digestion of the liver samples with nitric acid, copper is extracted as the salicylate—pyridine complex into chloroform and determined catalytically directly in the organic extract by addition of ethanolic solutions of sulphanilic acid, pyridine and hydrogen peroxide. Copper can be determined in the range from 10 to 350 ppm (dry weight basis) with a relative standard deviation of 5–15%. The method enables copper to be determined in liver biopsy samples of about 5 mg dry weight by use of a simple spectrophotometer, and can be used in diagnosis of Wilson's disease.

Möglichkeiten und Grenzen der Diagnose Kupferspeicherkrankheit aus Biopsiematerial der Leber

IntroductionThe morphological diagnosis of Wilson's disease cannot be made from the histological section alone and is aided by the quantitative analysis of copper in liver tissues. While, according to Sternlieb et al. (1968) , for this determination the liver tissue must be removed and processed under certain given conditions, in this investigation we examined the question as to whether it is possible to obtain and adequately reliable diagnosis using the tissue removed and fixed in formalin in accordance with conventional techniques, and whether this diagnosis is also possible in residual liver embedded in paraffin for histological examination. Material and methodsLiver tissue, obtained per needle biopsy or with the scalpel was examined with respect to its content of copper using the technique of atomic absorption spectrophotometry. Copper determinations in 107 cadaver livers were evaluated. In each of 10 autopsy cases with morphologically intact livers, a single specimen of liver tissue, removed with the scalpel, and also 2–10 needle biopsies from each right liver lobe were processed a) after simple drying b) after fixation in formalin and drying c) after fixation in formalin and embedding in Autotechnicon as for histological processing, followed by de-paraffining in xylol and acetone and drying. The tissue specimens were subjected to acid hydrolysis and then measured against and aqueous standard solution. In the same manner, the hepatic copper was determined in residual material taken out of paraplast block in 90 morphologically intact liver cylinders of all age groups. This material had been sent to the laboratory for histological examination. The values obtained were compared with cases of Wilson's disease that had been confirmed either clinically or at autopsy. Resultats and discussionThe average copper content in 97 biopsy material samples removed with the scalpel and subjected to multiple determinations was 3.2 mg of copper per 100 g dry weight. The tissue of the biopsy specimens obtained with untreated needles was examined using the same method at a considerably stronger dilution of the measured samples and was found to give 10 mg per 100 g of dry liver tissue. The figures for dry liver tissue fixed in formalin are only slightly higher than those for unfixed tissue. Thus it is established that the demonstration of copper with adequate diagnostic information content is possible even in tissue fixed in formalin. The demonstration of copper in residual material from liver cylinders embedded in paraffin provides no diagnostically useful results. The reason for this is largely to be found in the embedding technique commonly employed at our institute. In the case of biopsy material removed with the scalpel and embedded in paraffin, which has had no opportunity of being contaminated exogenously with copper, the diagnosis of disturbed copper metabolism can be made unequivocally.