Blue native-polyacrylamide gel electrophoresis is a powerful technique that enables the separation of intact multi-subunit complexes. However, positive identification of particular enzymes generally requires further separation in a second dimension on a denaturing polyacrylamide gel. Histochemical staining is widely used to demonstrate enzyme activities in tissues, including oxidative phosphorylation enzymes. In this report, we demonstrate that the two techniques can be combined to quantify in situ mitochondrial enzymes, separated on nondenaturing polyacrylamide gels. The method gives quantitative results with human skeletal muscle as well as heart that contains higher mitochondrial numbers. Comparison of muscle from patients with oxidative phosphorylation enzyme deficiencies, such as those of two riboflavin-responsive patients, before and after vitamin treatment, gives results in agreement with those obtained by analyzing the activity of the mitochondrial enzymes in muscle homogenates.

This study examined the relationship between quality of life (QOL) and dual diagnosis among patients in treatment for opioid dependence. The study sample includes 57 patients with opioid dependence alone (OD) and 41 with opioid dependence and a psychiatric axis-I disorder (DD), recruited in 2001 and 2004 at the Drug Addiction Services (SerT) of Bolzano and Pontedera (Italy). Participants were 73.5% males, with a mean age of 35.1 years (SD = 8.0). A comparison group of 45 healthy controls was also included. Assessments included a structured psychiatric interview (SCID) and a self-report quality of life assessment (WHOQOL-BREF). Patients with DD reported significantly (p < 0.05) poorer QOL in the physical and psychological domains as compared with patients with OD. Both groups of patients with and without DD showed significantly (p < 0.001) poorer QOL in the physical, psychological, and social domains with respect to healthy participants. The scores on the "relationship with environment" domain did not differ among OD, DD, and controls. The present study provides preliminary evidence that dual diagnosis is associated with poorer QOL and emphasizes the need to target treatment for the mental disorder concomitantly with the dependence problem in patients in treatment for opioid dependence.

Two unrelated adult males, aged 36 (patient 1) and 25 (patient 2) years, presented with subacute carnitine-deficient lipid storage myopathy that was totally and partly responsive to riboflavin supplementation in the two patients, respectively. Plasma acyl-carnitine and urinary organic acid profiles indicated multiple acyl coenzyme A dehydrogenase deficiency, which was mild in patient 1 and severe in patient 2. The activities of short-chain and medium-chain acyl coenzyme A dehydrogenases in mitochondrial fractions were decreased, especially in patient 2. This was in agreement with Western blotting results. Flavin-dependent complexes I and II were studied by immunoblotting and densitometric quantification of two-dimensional electrophoresis with comparable results. Complex I was present in normal amounts in both patients, whereas complex II was decreased only in the pretherapy muscle of patient 2. Flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) concentrations in muscle and isolated mitochondria, and the activity of mitochondrial FAD pyrophosphatase, showed that patient 1 had low levels of FAD (46%) and FMN (49%) in mitochondria, with a significant increase (P < 0.01) in mitochondrial FAD pyrophosphatase (273%) compared with controls. Patient 2 had similar low levels of FAD and FMN in both total muscle (FAD and FMN 22% of controls) and mitochondria (FAD 26%; FMN 16%) and normal activity of mitochondrial FAD pyrophosphatase. All of these biochemical parameters were either totally or partly corrected after riboflavin therapy.

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