Alterations in Skeletal Muscle Insulin Signaling DNA Methylation: A Pilot Randomized Controlled Trial of Olanzapine in Healthy Volunteers.

Antipsychotics are associated with severe metabolic side effects including insulin resistance; however, the mechanisms underlying this side effect are not fully understood. The skeletal muscle plays a critical role in insulin-stimulated glucose uptake, and changes in skeletal muscle DNA methylation by antipsychotics may play a role in the development of insulin resistance. A double-blind, placebo-controlled trial of olanzapine was performed in healthy volunteers. Twelve healthy volunteers were randomized to receive 10 mg/day of olanzapine for 7 days. Participants underwent skeletal muscle biopsies to analyze DNA methylation changes using a candidate gene approach for the insulin signaling pathway. Ninety-seven methylation sites were statistically significant (false discovery rate < 0.05 and beta difference between the groups of ≥10%). Fifty-five sites had increased methylation in the skeletal muscle of olanzapine-treated participants while 42 were decreased. The largest methylation change occurred at a site in the Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-Alpha (PPARGC1A) gene, which had 52% lower methylation in the olanzapine group. Antipsychotic treatment in healthy volunteers causes significant changes in skeletal muscle DNA methylation in the insulin signaling pathway. Future work will need to expand on these findings with expression analyses.

Comparison of SOX-10, HMB-45, and Melan-A in Benign Melanocytic Lesions.

PURPOSE: Different immunohistochemical stains are used in dermatopathology to stain melanocytes and diagnose benign and malignant melanocytic lesions. METHODS: SOX-10, HMB-45, and Melan-A immunohistochemical stains were used to assess 32 biopsy specimens with a histologic diagnosis of lentigo. The total number of melanocytes stained with each immunohistochemical stain was counted and an average count was obtained from two readings. RESULTS: Analysis of the data revealed a significant difference in staining melanocytes between these three immunostains (p=0.0010, ANOVA). SOX-10 stained 0.195 more melanocytes than HMB-45 (p=0.0026). Similarly, Melan-A stained 0.195 more melanocytes than HMB-45 (p=0.0011). However, the difference between SOX-10 and Melan-A was not statistically significant (p=0.9810). CONCLUSION: SOX-10 and Melan-A immunostaining stain more melanocytes than HMB-45. No significant difference was noted between Melan-A and SOX-10.

Surgical outcomes for esotropia in children with high accommodative convergence/accommodation ratio.

PURPOSE: To assess if high accommodative convergence/accommodation (AC/A) ratio impacts surgical outcomes in children with esotropia (ET), and evaluate the appropriate target angle in surgical dosing in the presence of high AC/A ratio. METHODS: A retrospective chart review identified patients who underwent primary bilateral medial rectus (BMR) recessions for ET. Patients were excluded if follow-up was ≤2 months. Basic demographic information, visual acuity, stereopsis, alignment, and target angle for surgery were collected. High AC/A was defined as ≥10 prism diopter (Δ) deviation at near compared to distance. Outcome parameters were near and distance deviations ≤10Δ within orthophoria, and/or stereopsis postoperatively. Yates' continuity correction, unpaired t-test, regression analysis, and one-way ANOVA were used. RESULTS: We identified 103 patients, 23 with high AC/A and 80 with normal AC/A, preoperatively. Mean age was 4.0 ± 2.5 years. Surgical success measured by postoperative alignment was 48% and 45% in the high AC/A and normal AC/A groups, respectively (P = 1.0). There was a statistically significant difference in preoperative near deviation between high AC/A and normal AC/A groups (P = 0.0015); however, there was no significant difference in preoperative distance deviation (P = 0.061). In addition, there was not a significant difference in preoperative or postoperative stereopsis between high AC/A and normal AC/A groups (P = 0.88 and P = 0.44, respectively). There was a significant difference in the normal AC/A and high AC/A groups when target angle was directed toward preoperative near deviation as determined by one-way ANOVA (F = 170.88, P < 0.0001 and F = 14.61, P = 0.0010, respectively). CONCLUSION: In the setting of ET treated with BMR recession, the presence of high AC/A does not affect surgical success as measured by alignment and stereopsis. In addition, when high AC/A is present, surgical dosing with a target angle toward near deviation was found to yield the best surgical outcomes in our patient population.

Skeletal muscle DNA methylation modifications and psychopharmacologic treatment in bipolar disorder.

Both severe mental illness and atypical antipsychotics have been independently associated with insulin resistance and weight gain. Altered regulation of skeletal muscle DNA methylation may play a role. We aimed to evaluate DNA methylation modifications in human skeletal muscle samples to further understand its potential role in the metabolic burden observed in psychiatric patients and psychopharmacologic treatment. Subjects were included in our study if they had a bipolar diagnosis and were currently treated with a mood stabilizer or atypical antipsychotic. A healthy control group free of psychiatric or physical disease was also included for comparisons. Anthropometric, BMI and hemoglobin A1C (HbA1C%) were measured. Fasting skeletal muscle biopsies were obtained and methylation levels of 5-methycytosine (5-mC), 5-hydroxymethylcytosine (5-hmC) and 5-formylcytosine (5-fC) were measured. Skeletal muscle global methylation of 5-mC and 5-fC were significantly higher in bipolar subjects compared to healthy controls. 5-mC was significantly higher in the AAP group compared to the mood stabilizer group. Significant correlations were observed between 5-fC methylation and HbA1C%. Our findings suggest that psychiatric disease and treatment may influence some methylation measures in the skeletal muscle of patients with bipolar disorder, which may be further influenced by medication treatment.

Association of Protein Kinase B (AKT) DNA Hypermethylation with Maintenance Atypical Antipsychotic Treatment in Patients with Bipolar Disorder.

STUDY OBJECTIVE: Atypical antipsychotics cause insulin resistance that leads to an increased risk of diabetes mellitus and cardiovascular disease. Skeletal muscle is the primary tissue for uptake of glucose, and its dysfunction is considered one of the primary defects in the development of insulin resistance. Protein kinase B (AKT) plays an important role in overall skeletal muscle health and glucose uptake into the muscle. The objective of this study was to measure AKT isoform-specific gene methylation differences in the skeletal muscle of patients with bipolar disorder treated with atypical antipsychotic or mood stabilizer maintenance therapy. DESIGN: Cross-sectional observational study. SETTING: Clinical research services center at an academic center. PATIENTS: Thirty patients with a confirmed diagnosis of bipolar disorder who were treated with either an atypical antipsychotic (16 patients) or mood stabilizer (14 patients) at a consistent dose for at least 3 months. INTERVENTIONS: A fasting skeletal muscle biopsy was performed in the vastus lateralis in each patient. Patients also underwent fasting blood sample collection and a standard 75-g oral glucose tolerance test. MEASUREMENTS AND MAIN RESULTS: Skeletal muscle DNA methylation near the promoter region for three genes, AKT1, AKT2, and AKT3, was measured by methylation-sensitive high-resolution melting. Gene methylation was analyzed based on atypical antipsychotic versus mood stabilizer maintenance therapy. Associations between gene methylation, insulin resistance, and glucose tolerance were also analyzed. In patients treated with atypical antipsychotics, AKT1 and AKT2 methylation was increased compared with patients treated with mood stabilizers (p=0.03 and p=0.02, respectively). In addition, for patients receiving atypical antipsychotics, a positive trend for AKT2 hypermethylation with increasing insulin resistance was observed, whereas for patients receiving mood stabilizers, a trend for decreased AKT2 methylation with increasing insulin resistance was observed. CONCLUSION: Overall, our findings suggest that the AKT gene is differentially methylated in the skeletal muscle of patients taking atypical antipsychotics or mood stabilizer maintenance therapy. These results may direct future approaches to reduce the harmful adverse effects of atypical antipsychotic treatment.

Association between haptoglobin gene and insulin resistance in Arab-Americans.

AIM: To analyze associations between variation in the HP gene and lipid and glucose-related measures in Arab-Americans. Secondary analyses were performed based on sex. PATIENTS & METHODS: Genomic DNA was extracted from samples obtained from a previous epidemiological study of diabetes in Arab-Americans. The HP 1 and 2 alleles were analyzed by polymerase chain reaction and gel electrophoresis. Associations were analyzed by linear regression. RESULTS & CONCLUSION: Associations were identified between the heterozygous haptoglobin 2-1 genotype and insulin resistance, fasting insulin and fasting c-peptide. The effect of sex did not remain significant after adjustment for relevant variables. HP genetic variation may have utility as a biomarker of insulin resistance and diabetes risk in Arab-Americans, however, future prospective studies are needed.