Uncoupling protein 2 (UCP2) is suggested to be a regulator of reactive oxygen species production in mitochondria. We performed a detailed study of brain injury, including regional and cellular distribution of UCP2 mRNA, as well as measures of oxidative stress markers following permanent middle cerebral artery occlusion in UCP2 knockout (KO) and wild-type (WT) mice. Three days post ischemia, there was a massive induction of UCP2 mRNA confined to microglia in the peri-infarct area of WT mice. KO mice were less sensitive to ischemia as assessed by reduced brain infarct size, decreased densities of deoxyuridine triphosphate nick end-labelling (TUNEL)-labelled cells in the peri-infact area and lower levels of lipid peroxidation compared with WT mice. This resistance may be related to the substantial increase of basal manganese superoxide dismutase levels in neurons of KO mice. Importantly, we found a specific decrease of mitochondrial glutathione (GSH) levels in UCP2 expressing microglia of WT, but not in KO mice after ischemia. This specific association between UCP2 and mitochondrial GSH levels regulation was further confirmed using lipopolysaccharide models of peripheral inflammation, and in purified peritoneal macrophages. Moreover, our data imply that UCP2 is not directly involved in the regulation of ROS production but acts by regulating mitochondrial GSH levels in microglia. Keywords: cerebral ischemic injury, glutathione, lipopolysaccharide, reactive oxygen species, superoxide dismutase, uncoupling protein 2. Uncoupling protein 2 (UCP2) (Fleury et al. 1997;Boss et al. 2000;Ricquier and Bouillaud 2000) a homologue of the brown adipose tissue-specific proton transporter UCP1, belongs to the mitochondrial anion carrier family that are present in the inner mitochondrial membrane (el Moualij et al. 1997). The UCP2 gene is expressed in most tissues (Fleury et al. 1997;Gimeno et al. 1997;Pecqueur et al. 2001) including brain (Richard et al. 1998). Whereas the main function of UCP1 in rodents is to produce heat by allowing

Rabbits were immunized with reduced glutathione (gamma-glutamyl-cysteinyl-glycine) coupled to bovine serum albumin by glutaraldehyde or a mixture of glutaraldehyde and formaldehyde. The antisera that were formed were tested qualitatively, by screening them against more than 50 amino acids and peptide conjugates that had been immobilized on cellulose discs (spot test), and quantitatively, by immunogold analysis of test conjugates that had been embedded in an epoxy resin. It was shown that the antisera selectively recognized the reduced and oxidized forms of glutathione and that they did not exhibit any significant crossreactivity with glutamate, cysteine, glycine, gamma-glutamyl-cysteine or cysteinyl-glycine. Immunocytochemistry of Vibratome sections of rat cerebellum suggested that glutathione occurs in glial cells as well as in neurons. This was confirmed by electron microscopic, immunogold cytochemistry of tissue from rat cerebellum that had been freeze-substituted and embedded in Lowicryl under low temperature. Gold particles were concentrated over Golgi epithelial cells and perivascular glial processes, but also occurred over several types of neuronal profile including Purkinje and granule cell bodies, and mossy fibre terminals. At the subcellular level, glutathione-like immunoreactivity was found in the cytoplasmic matrix, mitochondria and nuclei. The immunolabelling intensity was strongly reduced in animals that had been pretreated with buthionine sulphoximine, which is known to depress the level of glutathione by inhibiting gamma-glutamyl-cysteine synthetase. The availability of antisera to glutathione is likely to further our understanding of the physiological and pathophysiological roles of this tripeptide.

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BackgroundThe prevalence of non-communicable diseases (NCDs) is increasing worldwide, also among children. Information about primary prevention of NCD’s is increasing; however, convincing strategies among children is needed. The present paper describes the design and methods in the Health Oriented Pedagogical Project (HOPP) study. The main objective is to evaluate the effects of a school-based physical activity intervention program on cardio-metabolic risk factors. Secondary objectives include assessment of physical, psychological and academic performance variables.MethodsThe HOPP study is a 7 years longitudinal large-scale controlled intervention in seven elementary schools (n = 1545) with two control schools (n = 752); all aged 6–11 years at baseline. The school-based physical activity intervention program includes an increase in physical activity (PA) of 225 min/week as an integrated part of theoretical learning, in addition to the curriculum based 90 min/week of ordinary PA. Primary outcomes include cardio-metabolic risk factors measured as PA level, BMI status, waist circumference, muscle mass, percent fat, endurance test performance, total serum cholesterol, high-density lipoprotein (HDL), non-HDL, micro C-reactive protein (mCRP) and long-term blood sugar (HbA1c). In addition, secondary outcomes include anthropometric growth measures, physical fitness, quality of life (QoL), mental health, executive functions, diet and academic performance.DiscussionHOPP will provide evidence of effects on cardio-metabolic risk factors after a long-term PA intervention program in elementary schoolchildren. School-based PA intervention programs may be an effective arena for health promotion and disease prevention.Trial registrationThe study is registered in Clinical trials (ClinicalTrials.gov Identifier: NCT02495714) as of June 20th – 2015, retrospectively registered. The collection of baseline values was initiated in mid-January 2015.

The results may be used as reference values for handgrip strength in 6-12-year-old children of both sexes. Handgrip strength may not be used as a screening tool for cardiometabolic risk factors in pre-pubertal children.

The aim of the present study was to elucidate the distribution of glutathione immunoreactivity in the normal hippocampus and after kainate-induced neuronal injury. A specific antibody was used that recognizes both the reduced (GSH) and oxidized (GSSG) forms of glutathione. Immunoreactivity to glutathione was observed in neurons, but few immunolabeled glial cells were observed in the normal hippocampus. After kainate injection, a decrease in glutathione immunoreactivity was observed in pyramidal neurons from as early as 1 day after injection. In contrast, dense staining to glutathione was observed in large numbers of reactive astrocytes at 3 days to 6 weeks after kainate injection. This suggests upregulation of glutathione synthesis in these cells. One possibility is that the high content of glutathione is protective to reactive astrocytes. Another possibility is that the high glutathione concentration in reactive astrocytes may be protective to neurons around the glial scar.

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