厚生労働科学研究費補助金（難治性疾患克服研究事業） 「Menkes 病・occipital horn 症候群の実態調査、早期診断基準確立、治療法開発に関する研究」 平成23年度 総括・分担研究報告書

厚生労働科学研究費補助金（難治性疾患克服研究事業） 「Menkes 病・occipital horn 症候群の実態調査、早期診断基準確立、治療法開発に関する研究」 平成23年度 総括・分担研究報告書（page 18/118）[厚生労働科学研究費補助金（難治性疾患克服研究事業） 「Menkes 病・occipital horn 症候群の実態調査、早期診断基準確立、治療法開発に関する研究」 平成23年度 総括・分担研究報告書]

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Assay for Cytochrome c oxidase activityMitochondria were isolated from tissue samples immediately after sacrifice using theMitochondria Isolation Kit for Tissue (Pierce, Rockford, IL). Cytochrome c ox....

Assay for Cytochrome c oxidase activityMitochondria were isolated from tissue samples immediately after sacrifice using theMitochondria Isolation Kit for Tissue (Pierce, Rockford, IL). Cytochrome c oxidase activity in themitochondrial solution was determined using the Cytochome c Oxidase Assay Kit (Sigma-Aldrich,St. Louis, MO). Protein concentration was determined using the Pierce BCA Protein Assay Kit(Pierce, Rockford, IL).Measurement of catecholamine and enzyme concentrationsCerebrum and cerebellum were homogenized in 0.4 N perchloric acid and centrifuged at4°C (12,000 g, 5 min). Catecholamines, including dopamine, noradrenaline, and adrenaline, in thesupernatant were analyzed using a catecholamine autoanalyzer (HLC-8030; Toso Ind., Tokyo,Japan). Serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), bloodurea nitrogen (BUN), and creatinine were analyzed by Oriental Yeast Co., Ltd (Shiga, Japan).Statistical analysisData are presented as mean+standard error (SE). The SE is computed from known samplestatistics, and it provides an unbiased estimate of the standard deviation of the statistic, thus we usedSE. Difference in body weight changes between two groups was analyzed using two-way repeatedmeasure ANOVA. Differences in copper concentration, CCO activity and catecholamine betweentwo groups were analyzed using one-way repeated measure ANOVA (Tukey post hoc test);p<0.05 was considered significant.Results and discussionAs shown in Fig. 1, weight gain in treated macular mice was significantly higher than that ofcontrol macular mice after week 7. Fig. 2 shows the copper concentration in tissues and serum.Consistent with a previous report [12], control macular mice exhibited low copper concentrations in