Global DNA promoter methylation in frontal cortex of alcoholics and controls.

To determine if ethanol consumption and alcoholism cause global DNA methylation disturbances, we examined alcoholics and controls using methylation specific microarrays to detect all annotated gene and non-coding microRNA promoters and their CpG islands. DNA was isolated and immunoprecipitated from the frontal cortex of 10 alcoholics and 10 age and gender-matched controls then labeled prior to co-hybridization. A modified Kolmogorov-Smirnov test was used to predict differentially enriched regions (peaks) from log-ratio estimates of amplified vs input DNA. More than 180,000 targets were identified for each subject which correlated with >30,000 distinct, integrated peaks or high probability methylation loci. Peaks were mapped to regions near 17,810 separate annotated genes per subject representing hypothetical methylation targets. No global methylation differences were observed between the two subject groups with 80% genetic overlap, but extreme methylation was observed in both groups at specific loci corresponding with known methylated genes (e.g., H19) and potentially other genes of unknown methylation status. Methylation density patterns targeting CpG islands visually correlated with recognized chromosome banding. Our study provides insight into global epigenetic regulation in the human brain in relationship to controls and potentially novel targets for hypothesis generation and follow-up studies of alcoholism.

Plasma cytokine levels in children with autistic disorder and unrelated siblings.

BACKGROUND: The pathogenesis of autistic disorder (AD) is not clearly understood but genetic factors and the immune system have been implicated. Disturbed immunoglobulin levels and autoantibodies to neuronal elements have been reported in AD including cytokines encoded by genes involved with cell proliferation, migration and adhesion but there is a paucity of data comparing cytokine levels in children with AD and unrelated siblings without AD. METHODS: We analyzed 39 plasma cytokines in 99 well-characterized children with AD between 5 and 10 years of age and 40 age and gender matched healthy unrelated siblings without AD under the same clinical assessments, specimen processing and laboratory conditions. Multiplex sandwich immunoassays were used with the Luminex fluorescent-bead based platform. Log-transformed values of the 29 cytokines meeting laboratory criteria for inclusion were analyzed by analysis of covariance with a general linear model adjusting for diagnosis, gender, diagnosis by gender interaction effects, age and days of specimen handling. The Tukey-Kramer post hoc test was used to control for multiple comparisons. RESULTS: Eight of 29 cytokine levels analyzed were significantly lower in children with AD compared with unrelated siblings without the diagnosis of AD. Three of the cytokines are known to be involved with hematopoiesis and five with attraction of T-cells, natural killer cells and monocytes. CONCLUSIONS: Plasma cytokine levels representing chemokines involved in the T-helper cell immune system and hematopoiesis were lower in the children with AD compared with unrelated siblings without AD necessitating further studies to confirm immunological disturbances influencing hematopiesis and antibody production in the children with AD. Linking genes that encode immune related proteins and cytokines are important to study for their impact on critical periods of brain development and function.