Isoflurane disrupts excitatory neurotransmitter dynamics via inhibition of mitochondrial complex I.

BACKGROUND: The mechanisms of action of volatile anaesthetics are unclear. Volatile anaesthetics selectively inhibit complex I in the mitochondrial respiratory chain. Mice in which the mitochondrial complex I subunit NDUFS4 is knocked out [Ndufs4(KO)] either globally or in glutamatergic neurons are hypersensitive to volatile anaesthetics. The volatile anaesthetic isoflurane selectively decreases the frequency of spontaneous excitatory events in hippocampal slices from Ndufs4(KO) mice. METHODS: Complex I inhibition by isoflurane was assessed with a Clark electrode. Synaptic function was measured by stimulating Schaffer collateral fibres and recording field potentials in the hippocampus CA1 region. RESULTS: Isoflurane specifically inhibits complex I dependent respiration at lower concentrations in mitochondria from Ndufs4(KO) than from wild-type mice. In hippocampal slices, after high frequency stimulation to increase energetic demand, short-term synaptic potentiation is less in KO compared with wild-type mice. After high frequency stimulation, both Ndufs4(KO) and wild-type hippocampal slices exhibit striking synaptic depression in isoflurane at twice the 50% effective concentrations (EC50). The pattern of synaptic depression by isoflurane indicates a failure in synaptic vesicle recycling. Application of a selective A1 adenosine receptor antagonist partially eliminates isoflurane-induced short-term depression in both wild-type and Ndufs4(KO) slices, implicating an additional mitochondria-dependent effect on exocytosis. When mitochondria are the sole energy source, isoflurane completely eliminates synaptic output in both mutant and wild-type mice at twice the (EC50) for anaesthesia. CONCLUSIONS: Volatile anaesthetics directly inhibit mitochondrial complex I as a primary target, limiting synaptic ATP production, and excitatory vesicle endocytosis and exocytosis.

Obesogenic environments: Are neighbourhood environments that limit physical activity obesogenic?

The impact of obesogenic environments on adolescent health is poorly understood. This study examines if neighbourhood features related to physical activity are also related to unhealthy weight status. Adolescents (N=4587, age 15-17 years, 51.4% male) self-reported physical activity and neighbourhood perceptions. Trained researchers measured height and weight. Logistic regression identified if neighbourhood perceptions predicted overweight or obesity. Adolescents who reported convenient physical activity facilities were 2% less likely to be overweight/obese and 5% less likely to be obese, controlled for socio-demographic variables and clustering by school. Physical activity does not appear to directly influence or mediate the relationship between perceived convenient facilities and weight status.

General practitioners knowledge, practice and training requirements in relation to doping in sport.

This study examined General Practitioner's (GP) knowledge, practice and training requirements in relation to doping in sport in Ireland. All 2083 GPs on the Irish College of General Practitioners (ICGP) register received a postal questionnaire, yielding a 37% response rate (N=771, 63% male, average age 46.2 +/- 9SD, range 28-74 years). Results revealed that 14% (112) deemed their knowledge of doping agents to be good or very good, 12% (94) had completed specific training modules in doping or sport, and 24% (183) were connected with a specific sport as a team doctor/advisor. Over one in four (28%: 217) had been consulted for advice on doping in Sport, 33% (256) possessed the current list of prohibited substances, and 25% (190) knew of the Irish Sports Council's drug-testing procedures. The current initiatives to discourage doping in sport were felt to be ineffective, and although 92% (716) indicated that GPs had a role to play in the prevention of doping in sport, only 9% (66) felt adequately trained for such a role. There was overwhelming support for further training among GPs, although the most appropriate method of providing training is complex and requires strategic planning.

Quantitative analysis of associations between DNA hypermethylation, hypomethylation, and DNMT RNA levels in ovarian tumors.

How hypermethylation and hypomethylation of different parts of the genome in cancer are related to each other and to DNA methyltransferase (DNMT) gene expression is ill defined. We used ovarian epithelial tumors of different malignant potential to look for associations between 5'-gene region or promoter hypermethylation, satellite, or global DNA hypomethylation, and RNA levels for ten DNMT isoforms. In the quantitative MethyLight assay, six of the 55 examined gene loci (LTB4R, MTHFR, CDH13, PGR, CDH1, and IGSF4) were significantly hypermethylated relative to the degree of malignancy (after adjustment for multiple comparisons; P < 0.001). Importantly, hypermethylation of these genes was associated with degree of malignancy independently of the association of satellite or global DNA hypomethylation with degree of malignancy. Cancer-related increases in methylation of only two studied genes, LTB4R and MTHFR, which were appreciably methylated even in control tissues, were associated with DNMT1 RNA levels. Cancer-linked satellite DNA hypomethylation was independent of RNA levels for all DNMT3B isoforms, despite the ICF syndrome-linked DNMT3B deficiency causing juxtacentromeric satellite DNA hypomethylation. Our results suggest that there is not a simple association of gene hypermethylation in cancer with altered DNMT RNA levels, and that this hypermethylation is neither the result nor the cause of satellite and global DNA hypomethylation.

Inherited phosphofructokinase deficiency in an American cocker spaniel.

A 3-year-old female American Cocker Spaniel with a chronic hemolytic disorder and hemolytic crises was found to have M-type phosphofructokinase deficiency. This inherited erythroenzymopathy and myopathy is commonly diagnosed in English Springer Spaniels, but the family study of this Cocker Spaniel, although supporting an autosomal recessive mode of inheritance, did not reveal any English Springer Spaniel ancestors. Molecular genetic studies did, however, identify the same mutation in this dog as we previously reported in the English Springer Spaniel breed, suggesting that this mutation originated prior to the separation of these 2 breeds.

Esophageal deviation in four English Bulldogs.

Regurgitation, vomiting, and aspiration pneumonia in 4 young male English Bulldogs were attributed to esophageal deviation that was demonstrated by esophageal radiography. A genetic relationship among these dogs was not determined. Exploratory thoractomy, performed in 2 dogs, demonstrated an apparent compression of the esophagus by the left subclavian artery and brachiocephalic artery at the level of the 2nd and 3rd intercostal spaces. This apparent compression correlated with the roentgen location of esophageal deviation. Surgical redirection of the left subclavian artery resulted in immediate and long-lasting remission of clinical signs.

Anomalous portosystemic anastomoses associated with chronic hepatic insufficiency in six young dogs.

Chronic hepatic insufficiency due to anomalies of the portal venous system was diagnosed in 6 young dogs. The disorder was characterized by a variety of abnormal central nervous system signs or ascites, or both. Laboratory findings were characteristic of chronic, generalized hepatic dysfunction. The diagnosis was established by angiographic studies of the portal venous system. Of the 6 dogs, 3 died, 1 was euthanatized, and 2 are still alive and require medical management for ascites.