An impaired mitochondrial electron transport chain increases retention of the hypoxia imaging agent diacetylbis(4-methylthiosemicarbazonato)copper II

Radiolabeled diacetylbis(4-methylthiosemicarbazonato)copper^II [Cu^II(atsm)] is an effective positron-emission tomography imaging agent for myocardial ischemia, hypoxic tumors, and brain disorders with regionalized oxidative stress, such as mitochondrial myopathy, encephalopathy, and lactic acidosis with stroke-like episodes (MELAS) and Parkinson’s disease. An excessively elevated reductive state is common to these conditions and has been proposed as an important mechanism affecting cellular retention of Cu from Cu^II(atsm). However, data from whole-cell models to demonstrate this mechanism have not yet been provided. The present study used a unique cell culture model, mitochondrial xenocybrids, to provide whole-cell mechanistic data on cellular retention of Cu from Cu^II(atsm). Genetic incompatibility between nuclear and mitochondrial encoded subunits of the mitochondrial electron transport chain (ETC) in xenocybrid cells compromises normal function of the ETC. As a consequence of this impairment to the ETC we show xenocybrid cells upregulate glycolytic ATP production and accumulate NADH. Compared to control cells the xenocybrid cells retained more Cu after being treated with Cu^II(atsm). By transfecting the cells with a metal-responsive element reporter construct the increase in Cu retention was shown to involve a Cu^II(atsm)-induced increase in intracellular bioavailable Cu specifically within the xenocybrid cells. Parallel experiments using cells grown under hypoxic conditions confirmed that a compromised ETC and elevated NADH levels contribute to increased cellular retention of Cu from CuII(atsm). Using these cell culture models our data demonstrate that compromised ETC function, due to the absence of O2 as the terminal electron acceptor or dysfunction of individual components of the ETC, is an important determinant in driving the intracellular dissociation of Cu^II(atsm) that increases cellular retention of the Cu.

Lactation in Australian women : the effect of oral contraceptives on the vitamin and trace element content of breast milk

The aim of this project was to investigate the effects of oral contraceptives on the nutrient composition of breast milk. The design of the study also allowed the effects of stage of lactation and maternal diet on milk composition to be observed. A prospective study was designed to measure maternal dietary intake and vitamin and trace element concentration in milk and plasma. Vitamin A, ascorbic acid and iron, copper, zinc, manganese, selenium, cobalt, chromium, rubidium and caesium were measured. Two groups of women participated, oral contraceptive users and controls. Fasting milk and blood samples and 24-hour food records were collected from the women once a week for 20 weeks commencing 3-8 weeks post-partum, and 1-2 weeks before they began to take oral contraceptives. Fifteen women participated in the study; 5 took progestogen-only oral contraceptives, 1 took an oestrogen-progestogen oral contraceptive and 9 acted as controls. Progestogen-only oral contraceptives did not affect the milk or plasma concentration of the vitamins and trace elements measured. As only 1 subject took an oestrogen-progestogen preparation no conclusion could be drawn as to its effect. The mean milk and plasma concentration of all nutrients studied did not change significantly with the progression of lactation, with the exception of iron and zinc. The mean milk iron concentration was significantly higher at 16 weeks post-partum than at 8 and 23 weeks post-partum. The mean milk zinc concentration was significantly lower at 23 weeks post-partum than at 8 and 16 weeks post-partum. The infants1 mean estimated daily intakes of ascorbic acid and vitamin A from breast milk were above the U.S. and British Recommended Dietary Allowance for those vitamins. However, their mean estimated intakes of iron, zinc, copper, manganese and selenium were well below the U.S. recommendations. Effects of the maternal dietary intake on milk and plasma composition were variable. Implications of these findings have been discussed.

Blood mineral, trace-element and vitamin concentrations in Huacaya alpacas and Merino sheep grazing the same pasture

We aimed to determine whether the concentration of minerals and trace constituents in blood of Merino sheep and Huacaya alpacas grazing the same pasture differed with species and time of sampling. Blood samples and pasture samples were collected at frequent intervals over a period of 2 years for mineral and trace-nutrient assay. The concentration of the minerals and trace nutrients in the grazed pasture usually met the dietary needs of sheep at maintenance, apart from potassium, sulfur, cobalt and Vitamin E in occasional samples. Restricted maximum likelihood mixed model analysis indicated a significant (P < 0.001) species by month by year interaction for all blood constituents assayed, a significant (P < 0.05) species by coat shade interaction for plasma Vitamin D, E and B12 and a significant (P < 0.001) species by month by Vitamin D interaction for plasma phosphorus concentrations. In general, plasma calcium concentrations were greater in sheep than in alpacas but plasma magnesium concentrations were greater in alpacas than in sheep. There was no consistent difference between the two species in plasma phosphorus concentrations although low values were recorded in individual sheep and alpacas. Plasma Vitamin D concentrations were more responsive to increasing hours of sunlight in alpacas than they were in sheep. Sheep had consistently higher concentrations of plasma copper, zinc and Vitamin B12 and higher concentrations of blood selenium but lower concentrations of plasma selenium and Vitamin A, than did alpacas. No consistent difference was observed between the two species in plasma Vitamin E concentrations.