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Real-time quantitative PCR analysis of human perhiperal blood mononuclear cells following mind-body interventions

ABSTRACT: This study was the first randomized controlled trial to test if mind-body interventions can improve self-regulation in prisoners with different personality disorders, using a combination of gene expression, neural, and behavioral measures.Thirty prisoners with personality disorders who score high on psychopathy were assigned to a mindfulness intervention (n=10), a yoga intervention (n=10), or a wait-list control group (n=10) using stratified random sampling. Both mindfulness and yoga interventions were held at the same time and lasted three hours per day on five consecutive days. At baseline and after the intervention, we measured inflammation-related gene expression; resting state brain activit with electroencephalography (EEG); risk-taking and attention with cognitive tasks; event-related potentials (ERPs) related to the attention task; and stress, emotion regulation and mindfulness with questionnaires. We expected that both yoga and mindfulness would improve self-regulation (i.e., executive attention, emotion regulation and self-awareness), reduce stress and risk-taking behavior, downregulate inflammatory-related gene expression and increase alpha and theta power. Blood was collected for the analysis of changes in gene expression of 38 genes related to inflammation and the immune system (IL-1 signaling pathway). Venipuncture samples were collected into 10ml EDTA tubes, lysed and frozen at -80°C. Peripheral blood mononuclear cells were isolated by density gradient centrifugation. Ribonucleic acid (RNA) was extracted (QiaAmp RNA Blood Mini kit; Qiagen) and tested for suitable mass and integrity (NanoDrop One, Acclaro Sample Intelligence; Thermo Scientific). RNA was then converted to fluorescent cRNA for hybridization to IL-1 signaling pathway plate H96 (Bio-Rad Labs Ltd, UK). Gene expression data were expressed as cycle threshold (CT) values by calculating 2−ΔΔCT, and referencing to the geomean of all genes because endogenous control genes that are supposed to have constant levels of expression were not stable in this case. After normalizing 2−ΔΔCT to the wait-list control group median, the data were imported into SPSS and analyzed by group allocation based on change scores of each normalized 2−ΔΔCT. Intent-to-treat (ITT) principles were used for all the analyses. Post-intervention missing data was handled by running multiple imputations. A p-value of <0.05 was considered significant and Bonferroni correction was implemented to adjust for multiple comparisons (i.e., <.02 was considered significant in individual 2-group comparisons). The data were analyzed with the IBM SPSS 24.0 software (Statistical Package for Social Sciences, SPSS Inc., 237 Chicago, IL). The effectiveness of interventions was tested by calculating changes scores and running non-parametric Kruskal-Wallis test for K independent samples because the assumptions for ANOVA were not met. If there was a significant difference detected, it was followed with Mann-Whitney U test 240 and Kolmogorov-Smirnov Z test to determine specific group differences. Additionally, eta squared was used as a measure of effect size to determine the proportion of variability in outcomes that can be explained by group membership.We found no significant changes in gene expression or any other measures.

ORGANISM(S): Homo sapiens

PROVIDER: GSE134703 | GEO | 2020/10/12

REPOSITORIES: GEO

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Project description:This study is a collaboration between the Center for Applied Genomics (CAG) at Children's Hospital of Philadelphia (CHOP) and the Brain Behavior Laboratory at the University of Pennsylvania (Penn). The cohort consists of youths aged 8-21 years who consulted the CHOP network and volunteered to participate in genomic studies of complex pediatric disorders. All participants underwent clinical assessment, including a neuropsychiatric structured interview and review of electronic medical records. They were also administered a neuroscience based computerized neurocognitive battery (CNB) and a subsample underwent neuroimaging. These are described separately below. Clinical Testing: <ul><li>GOASSESS, a computerized, structured screener developed from a modified version of the Kiddie-Schedule for Affective Disorders and Schizophrenia (K-SADS, Kaufman et al. 1997, PMID: <a href="https://pubmed.ncbi.nlm.nih.gov/9204677/" target="_blank">9204677</a>). Components of the interview include a timeline of life events, demographics and medical history, Global Assessment of Functioning, and Interviewer Observations.</li></ul> <ul><li>A psychopathology symptom and criterion-related assessment of mood disorders (depression, mania/hypomania), anxiety disorders (overanxious/generalized-, separation-, social-anxiety, specific phobia, panic disorder, agoraphobia, obsessive compulsive disorder, post-traumatic stress disorder), behavioral disorders (attention deficit hyperactivity disorder, oppositional defiant disorder, conduct disorder), psychosis spectrum (psychosis and prodromal symptoms), eating disorders, suicidal thinking and behavior, and treatment history. This is also based on K-SADS.</li></ul> <ul><li>An abbreviated Family Interview for Genetics Studies (FIGS) to assess major domains of psychopathology in the proband's first-degree relatives.</li></ul> Computerized Neurocognitive Battery: The CNB, developed for large-scale studies, yields measures of accuracy and speed for domains of executive-control functions (abstraction, attention, working memory), episodic memory (verbal, facial, spatial), complex cognitive processing (language reasoning, nonverbal reasoning, spatial processing), social cognition (emotion identification, emotion intensity differentiation, age differentiation) and sensorimotor and motor speed. The following neurobehavioral domains were assessed: <ul><li>Penn Conditional Exclusion Test is a measure of abstraction and concept formation. Participants decide which of 4 objects does not belong with the other 3, based on one of three sorting principles, which change. Feedback is used.</li></ul> <ul><li>Attention: The Penn Continuous Performance Test. Participants respond to a set of 7-segment displays whenever they form a digit or letter.</li></ul> <ul><li>Working Memory: The Letter N-back Test displays sequences of uppercase letters with a stimulus duration of 500 ms (ISI 2,500 ms.) In the 0-back condition, participants respond to a single target (i.e., X). In the 1-back condition they respond if the letter is identical to that preceding it. In the 2-back condition, they respond if the letter is identical to that presented two trials back.</li></ul> <ul><li>Verbal Memory: The Penn Word Memory Test presents 20 target words that are then mixed with 20 distracters equated for frequency, length, concreteness and low imageability. A 20 min delayed recall procedure is also administered.</li></ul> <ul><li>Face Memory: The Penn Face Memory Test presents 20 digitized faces that are then mixed with 20 distracters equated for age, gender and ethnicity. The procedure is repeated at 20 min delay.</li></ul> <ul><li>Spatial Memory: The Visual Object Learning Test uses Euclidean shapes as stimuli with the same paradigm as the word and face.</li></ul> <ul><li>Language and Analogical Reasoning: The Penn Verbal Reasoning consists of verbal analogy problems.</li></ul> <ul><li>Spatial Processing: Penn Line Orientation Test presents two lines at an angle, and participants click on a button that makes one line rotate until it has the same angle as the other.</li></ul> <ul><li>Emotion Processing: Facial displays of 4 emotions (Happy, Sad, Anger, Fear) and Neutral faces, 8 each, are presented and the subject identifies the emotion in a multiple-choice format. The facial stimuli are balanced for gender, age, and ethnicity.</li></ul> <ul><li>Sensory-motor Processing Speed: The task requires moving the mouse and clicking on a green square that disappears after the click. The square gets increasingly small and appears in unpredictable locations.</li></ul> Neuroimaging Protocol: Studies were performed at Penn using a Siemens Trio (Erlangen, Germany) 3T scanner equipped with 40mT/m gradients and 200 mT/m/s slew-rates. RF transmission utilized a quadrature body-coil, and reception a 12-channel head coil optimized for parallel imaging. Total image acquisition time was about 45 min. Structural Imaging: The T1-weighted protocol utilized a 3D, inversion-recovery, and magnetization-prepared rapid acquisition gradient echo. Relevant imaging procedures include: <ul><li>Structural magnetic imaging</li></ul> <ul><li>Diffusion tensor imaging</li></ul> <ul><li>ASL perfusion</li></ul> <ul><li>BOLD fMRI</li></ul> Neuroimaging tasks: <ul><li>Fractal N-Back Task of Spatial Working Memory</li></ul> <ul><li>Face Emotion Identification Task</li></ul> Neuroimages: The current data release includes over 9700 MRI images that may be downloaded through Authorized Access.

Project description:Lifestyle intervention can improve insulin sensitivity in obese youth yet few studies have examined the biological mechanisms underlying improvements. Therefore, the purpose of this study was to explore biological pathways associated with intervention-induced improvements in insulin sensitivity. Fifteen (7M/8F) overweight/obese (BMI percentile=96.3M-BM-11.1) Latino adolescents (15.0M-BM-10.9 years) completed a 12-week lifestyle intervention that included weekly nutrition education and 180 minutes of moderate-vigorous exercise per week. Insulin sensitivity, estimated by an oral glucose tolerance test and the Matsuda Index, increased 29.2% post intervention (2.4M-BM-10.3 to 3.1M-BM-10.3, p=0.01). Global microarray analysis profiling from whole blood was performed to examine changes in gene expression and to explore biological pathways that were significantly changed in response to the intervention. A total of 1,459 probes corresponding to mRNA transcripts (717 up, 742 down) were differentially expressed with a fold changeM-bM-^IM-%1.2 and P<0.05. Among the genes identified were hexokinase 3 (HK3), ATPase, H+ transporting V0 subunit e2 (ATPV0E), and sterol regulatory element binding transcription factor 1 (SREBF1), and endothelial cell adhesion molecule (ESAM). There were 8 pathways identified that met the criteria for significance, including insulin signaling, type 1 diabetes, and glycerophospholipid metabolism. Participants that increased insulin sensitivity exhibited five times the number of significant genes altered compared to non-responders (1,144 vs. 230). These findings offer insight into the molecular mechanisms underlying health improvements among high-risk Latino youth. Lifestyle interventions may contribute to improved insulin sensitivity through pathways related to insulin signaling and immune response. Further, genetic factors may mediate response to lifestyle intervention. Fifteen (7M/8F) overweight/obese Latino Youth Whole blood RNA samples evaluated pre and post intervention.

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Real-time quantitative PCR analysis of human perhiperal blood mononuclear cells following mind-body interventions

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