Art therapy masks reflect emotional changes in military personnel with PTSS.

Among disabling post-traumatic stress symptoms (PTSS) are irritability, aggressive behavior, distressing memories and general impaired cognition and negative mood. Art therapy interventions, including mask-making, can potentially alleviate these symptoms. We tested the hypothesis that art conveys emotions and predicted that blinded viewers would be able to perceive changes in theoretically derived emotional profiles expressed in art made by military personnel with PTSS from the onset to the end of therapy. Five service members and veterans exhibiting PTSS were enrolled in an 8-session art therapy protocol, during which they artistically transformed papier-mâché masks at the beginning and end of the protocol. We found that blinded viewers without knowledge of the masks' creation stage (onset or end of therapy) read initial masks as conveying more negative emotions (e.g., angry, upset, and challenged) and later masks as conveying more positive emotions (calm and pleasure). Based on the assessments from the blinded evaluators, we infer the emotional transition experienced by the participants was expressed in the masks. In an exploratory arm of the study, we also found that viewers were better able to empathize with the negative emotions experienced by participants with PTSS when asked to explicitly take their perspective.

Preventing Adverse Outcomes for Bereaved Youth: Indirect Effects From a Randomized Trial of the Family Bereavement Program on Fear of Abandonment, Grief, and Mental Health.

OBJECTIVES: We investigated whether the self-system belief of fear of abandonment mediated the effects of intervention-induced change in 2 protective factors-positive parenting and adaptive coping-and one risk factor-stressful events-on youth mental health problems and maladaptive grief. This study extends prior research on fear of abandonment in youth who experience parental death by examining pathways through which a program reduced fear of abandonment and, in turn, affected subsequent pathways to child mental health problems in the context of a randomized experiment. METHODS: This is a secondary data analysis study. We used data from the 4-wave longitudinal 2-arm parallel randomized controlled trial of the Family Bereavement Program conducted between 1996 and 1999 in a large city in the Southwestern United States. The sample consisted of 244 offspring between 8 and 16 at the pretest. They were assessed again at posttest, 11-month follow-up, and 6-year follow-up. Offspring, caregivers, and teachers provided data. RESULTS: Mediation analyses indicated that intervention-induced reductions in stressful events were prospectively associated with a lower fear of abandonment. For girls, fear of abandonment was related to self-reported maladaptive grief and teacher-reported internalizing problems 6 years later. CONCLUSIONS: This study extends prior research on the relation between intervention-induced changes in risk and protective factors and improvements in outcomes of bereaved youth. The findings support the reduction of stressful events as a key proximal target of prevention programs for bereaved children.

Enhancing the Focus: How Does Parental Incarceration Fit into the Overall Picture of Adverse Childhood Experiences (ACEs) and Positive Childhood Experiences (PCEs)?

Despite the five million children in the U.S. with an incarcerated parent, there is limited research on risk and protective factors for this population. We analyzed data from the National Survey for Children's Health (2018) to: (1) examine associations among parental incarceration and other adverse childhood experiences (ACEs), (2) characterize the association between parental incarceration and youth mental health outcomes, (3) examine differences in positive childhood experiences (PCEs; collective socialization, community engagement, neighborhood amenities, and family problem solving) by parental incarceration status, (4) examine whether PCEs were protective against mental health problems and if there was an interaction with parental incarceration status, and (5) examine the interaction between PCEs, parental incarceration, and ACEs on mental health problems. Results revealed that children with incarcerated parents had higher odds of experiencing other ACEs, higher odds of having mental health problems, and experienced fewer PCEs compared to children without incarcerated parents. Further, although PCEs were associated with a lower odds of mental health problems for both children with and without incarcerated parents, they did not mitigate the negative impact of parental incarceration on mental health outcomes. While PCEs attenuated the association between ACEs and mental health, parental incarceration status did not significantly moderate the interaction. These results highlight vulnerabilities and potential protective factors for children with incarcerated parents and have important implications for the development of multilevel intervention strategies that seek to promote resilience and reduce risk for this population.

Effects of a preventive parenting intervention for bereaved families on the intergenerational transmission of parenting attitudes: Mediating processes.

This study evaluated whether the Family Bereavement Program (FBP), a prevention program for parentally bereaved families, improved parenting attitudes toward parental warmth and physical punishment in young adult offspring 15 years after participation and identified mediational cascade pathways. One hundred fifty-six parents and their 244 offspring participated. Data were collected at pretest (ages 8-16), posttest, and six- and 15-year follow-ups. Ethnicity of offspring was: 67% non-Hispanic Caucasian, 16% Hispanic, 7% African American, 3% Native American, 1% Asian or Pacific Islander, and 6% other; 54% were males. There was a direct effect of the FBP on attitudes toward physical punishment; offspring in the FBP had less favorable attitudes toward physical punishment. There were also indirect effects of the FBP on parenting attitudes. The results supported a cascade effects model in which intervention-induced improvements in parental warmth led to fewer externalizing problems in adolescence/emerging adulthood, which in turn led to less favorable attitudes toward physical punishment. In addition, intervention-induced improvements in parental warmth led to improvements in anxious romantic attachment in mid-to-late adolescence/emerging adulthood, which led to more favorable attitudes toward parental warmth in emerging/young adulthood. These findings suggest that the effects of relatively brief prevention programs may persist into subsequent generations.

Does Binge-Watching eHealth Intervention Content Impact Outcomes?

PURPOSE: Advancements in technology have made it possible to deliver parenting interventions online, known as eHealth interventions. Little is known about the rate at which parents participate in eHealth interventions, characteristics of parents who watch eHealth interventions at an accelerated pace (i.e., binge-watching), and if binge-watching impacts intervention outcomes. METHODS: The sample included 142 Hispanic parents who were randomly assigned to an eHealth family-based intervention and completed 100% of eight online, prerecorded and self-paced video group sessions delivered across 12 weeks. We examined baseline predictors (parent sociodemographic characteristics, report of child's externalizing behaviors, and family functioning) of watching group sessions in two weeks or less (n = 23, 16.2%). Using latent growth curve modeling, we tested the impact of binge-watching on the trajectory of adolescent drug use, condomless sex, and depressive symptoms across 36 months. We also examined the impact of binge-watching on changes in family functioning from baseline to 6 months postbaseline. RESULTS: Parents with high levels of education and of children with attention problems were more likely to binge-watch. Conversely, parents of children with conduct disorder symptoms were less likely to binge-watch. The trajectory of depressive symptoms increased for adolescents with parents who binge-watched the intervention, but the trajectory of condomless sex decreased. There was no impact on drug use. Binge-watching was also associated with decreases in parental monitoring. DISCUSSION: The findings of this study have implications for eHealth interventions; the pace that parents watch eHealth interventions may subsequently impact adolescent outcomes, such as condomless sex and depressive symptoms.

The Prospective Effects of Caregiver Parenting on Behavioral Health Outcomes for Children with Incarcerated Parents: a Family Resilience Perspective.

Rates of parental incarceration in the USA have increased dramatically over the past four decades. The Adverse Childhood Experiences study identified parental incarceration as one of several risk factors related to multiple health outcomes during childhood and adulthood. Parents and other caregivers are widely regarded as sources of resilience for children experiencing adversity, yet few studies have examined caregivers' parenting practices as sources of resilience for children with incarcerated parents. This study used secondary data from a longitudinal randomized controlled trial of the prison-based parent management training program Parenting Inside Out (PIO). Specifically, it included 149 caregivers (i.e., the non-incarcerated parent, extended family member, or other adult who provides the day-to-day caretaking of a child during parental incarceration) of children aged 2-14 years whose incarcerated parents were randomly assigned to receive PIO or the control condition. Path analysis was used to examine associations between caregivers' parenting, social support, self-efficacy, and change in child internalizing and externalizing symptoms across a 6-month period. Direct effects of caregivers' parenting were found on improvements in child behavioral health from baseline (conducted when the parent was incarcerated) to the 6-month follow-up (conducted after most parents had been released). Indirect effects were found for caregiver social support and self-efficacy. The findings highlight the importance of caregivers' adaptive parenting as a protective resource for children who experience parental incarceration and have implications for the design of preventive interventions for this underserved population.

Immune-Related Genomic Schizophrenic Subtyping Identified in DLPFC Transcriptome.

Well-documented evidence of the physiologic, genetic, and behavioral heterogeneity of schizophrenia suggests that diagnostic subtyping may clarify the underlying pathobiology of the disorder. Recent studies have demonstrated that increased inflammation may be a prominent feature of a subset of schizophrenics. However, these findings are inconsistent, possibly due to evaluating schizophrenics as a single group. In this study, we segregated schizophrenic patients into two groups ("Type 1", "Type 2") by their gene expression in the dorsolateral prefrontal cortex and explored biological differences between the subgroups. The study included post-mortem tissue samples that were sequenced in multiple, publicly available gene datasets using different sequencing methods. To evaluate the role of inflammation, the expression of genes in multiple components of neuroinflammation were examined: complement cascade activation, glial cell activation, pro-inflammatory mediator secretion, blood-brain barrier (BBB) breakdown, chemokine production and peripheral immune cell infiltration. The Type 2 schizophrenics showed widespread abnormal gene expression across all the neuroinflammation components that was not observed in Type 1 schizophrenics. Our results demonstrate the importance of separating schizophrenic patients into their molecularly defined subgroups and provide supporting evidence for the involvement of the immune-related pathways in a schizophrenic subset.

Chronic Traumatic Encephalopathy in the Brains of Military Personnel.

BACKGROUND: Persistent neuropsychiatric sequelae may develop in military personnel who are exposed to combat; such sequelae have been attributed in some cases to chronic traumatic encephalopathy (CTE). Only limited data regarding CTE in the brains of military service members are available. METHODS: We performed neuropathological examinations for the presence of CTE in 225 consecutive brains from a brain bank dedicated to the study of deceased service members. In addition, we reviewed information obtained retrospectively regarding the decedents' histories of blast exposure, contact sports, other types of traumatic brain injury (TBI), and neuropsychiatric disorders. RESULTS: Neuropathological findings of CTE were present in 10 of the 225 brains (4.4%) we examined; half the CTE cases had only a single pathognomonic lesion. Of the 45 brains from decedents who had a history of blast exposure, 3 had CTE, as compared with 7 of 180 brains from those without a history of blast exposure (relative risk, 1.71; 95% confidence interval [CI], 0.46 to 6.37); 3 of 21 brains from decedents with TBI from an injury during military service caused by the head striking a physical object without associated blast exposure (military impact TBI) had CTE, as compared with 7 of 204 without this exposure (relative risk, 4.16; 95% CI, 1.16 to 14.91). All brains with CTE were from decedents who had participated in contact sports; 10 of 60 contact-sports participants had CTE, as compared with 0 of 165 who had not participated in contact sports (point estimate of relative risk not computable; 95% CI, 6.16 to infinity). CTE was present in 8 of 44 brains from decedents with non-sports-related TBI in civilian life, as compared with 2 of 181 brains from those without such exposure in civilian life (relative risk, 16.45; 95% CI, 3.62 to 74.79). CONCLUSIONS: Evidence of CTE was infrequently found in a series of brains from military personnel and was usually reflected by minimal neuropathologic changes. Risk ratios for CTE were numerically higher among decedents who had contact-sports exposure and other exposures to TBI in civilian life than among those who had blast exposure or other military TBI, but the small number of CTE cases and wide confidence intervals preclude causal conclusions. (Funded by the Department of Defense-Uniformed Services University Brain Tissue Repository and Neuropathology Program and the Henry M. Jackson Foundation for the Advancement of Military Medicine.).

Differential Sphingosine-1-Phosphate Receptor-1 Protein Expression in the Dorsolateral Prefrontal Cortex Between Schizophrenia Type 1 and Type 2.

Understanding the etiology and treatment approaches in schizophrenia is challenged in part by the heterogeneity of this disorder. One encouraging progress is the growing evidence that there are subtypes of schizophrenia. Recent in vitro findings of messenger ribonucleic acid (mRNA) gene expression on postmortem dorsolateral prefrontal cortex (DLPFC) showed that schizophrenia has two subtypes, those with a relatively normal DLPFC transcriptome (Type 1) and those with differentially expressed genes (Type 2). Sphingosine-1-phosphate receptor-1 (S1PR1) is one of the genes that was highly upregulated in Type 2 compared to Type 1 and controls. The impact of that finding is limited because it only can be confirmed through analysis of autopsy tissue, and the clinical characteristics such as symptoms severity or illness duration except for cause of death was not available from that Medical Examiner based autopsy study. However, S1PR1 has great potential because it is a target gene that can be accessed via positron emission tomography (PET) in vivo using specific radioligands (starting with [11C]CS1P1) successfully developed at our center in human brain imaging. As a preliminary study to validate this PET target in schizophrenia, S1PR1 protein expression was assessed by receptor autoradiography (ARG) using [3H]CS1P1 and immunohistochemistry (IHC) in the DLPFC from patients with schizophrenia classified as Type 1 or Type 2 based on their DLPFC transcriptomes and from controls. Our analyses demonstrate that ARG S1PR1 protein expression is significantly higher in Type 2 compared to Type 1 (p < 0.05) and controls (p < 0.05), which was consistent with previous mRNA S1PR1. These findings support the possibility that PET S1PR1 can be used as a future imaging biomarker to distinguish these subgroups of schizophrenic patients during life with obvious implications for both patient management and the design of clinical trials to validate novel pharmacologic therapies.

Deep transcriptome sequencing of subgenual anterior cingulate cortex reveals cross-diagnostic and diagnosis-specific RNA expression changes in major psychiatric disorders.

Despite strong evidence of heritability and growing discovery of genetic markers for major mental illness, little is known about how gene expression in the brain differs across psychiatric diagnoses, or how known genetic risk factors shape these differences. Here we investigate expressed genes and gene transcripts in postmortem subgenual anterior cingulate cortex (sgACC), a key component of limbic circuits linked to mental illness. RNA obtained postmortem from 200 donors diagnosed with bipolar disorder, schizophrenia, major depression, or no psychiatric disorder was deeply sequenced to quantify expression of over 85,000 gene transcripts, many of which were rare. Case-control comparisons detected modest expression differences that were correlated across disorders. Case-case comparisons revealed greater expression differences, with some transcripts showing opposing patterns of expression between diagnostic groups, relative to controls. The ~250 rare transcripts that were differentially-expressed in one or more disorder groups were enriched for genes involved in synapse formation, cell junctions, and heterotrimeric G-protein complexes. Common genetic variants were associated with transcript expression (eQTL) or relative abundance of alternatively spliced transcripts (sQTL). Common genetic variants previously associated with disease risk were especially enriched for sQTLs, which together accounted for disproportionate fractions of diagnosis-specific heritability. Genetic risk factors that shape the brain transcriptome may contribute to diagnostic differences between broad classes of mental illness.

Longitudinal Effects of PostDivorce Interparental Conflict on Children's Mental Health Problems Through Fear of Abandonment: Does Parenting Quality Play a Buffering Role?

In a sample of 559 children (ages 9-18), researchers investigated whether: (a) fear of abandonment mediated the association between postdivorce interparental conflict (IPC) and mental health problems, and (b) parent-child relationship quality moderated the association between IPC and fear of abandonment. Mediation analyses indicated that pretest IPC predicted fear of abandonment 3 months later, which then predicted child- and teacher-reported mental health problems 10 months later. The hypothesized protective effect of a high-quality parent-child relationship was not observed. IPC predicted fear of abandonment for all children, except for those with low- and moderate-quality father-child relationships, for whom IPC was not significantly related to fear of abandonment. Findings highlight the need to optimize child coping programs and improve parenting-after-divorce programs to reduce IPC.

Parenting time, parenting quality, interparental conflict, and mental health problems of children in high-conflict divorce.

Despite widespread acknowledgment that "frequent, continuing, and meaningful" (Pruett & DiFonzo, 2014) time with both parents is beneficial for children from divorced or separated families, and that interparental conflict (IPC) is associated with increased child mental health problems, the joint effects of parenting time (PT), parenting quality (PQ), and IPC on children's mental health problems are less clear. The current study integrates two theoretical models in multiple mediator analyses to test indirect effects of mothers' and fathers' PQ and IPC to explain the association between PT and children's mental health problems within the same model. Participants were children aged 9-18 years (N = 141) who had one or both parents participate in a randomized comparative effectiveness trial of a court-based prevention program for high-conflict divorcing or separating families. Data were collected at pretest and 9-month follow-up. Analyses revealed an indirect effect in which fathers' PQ mediated the association between PT and child internalizing problems both concurrently and 9 months later. There were no significant indirect effects involving IPC. Analyses indicated a significant quadratic relation between PT and fathers' PQ, suggesting that although more PT is associated with better father-child relationships, there is a point beyond which more time is not related to a better relationship. We discuss the study findings, research limitations, and implications for public policy. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Correction: DLPFC transcriptome defines two molecular subtypes of schizophrenia.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Long-term Effects of a Parenting Preventive Intervention on Young Adults' Attitudes Toward Divorce and Marriage.

This study examined whether the New Beginnings Program (NBP), a parenting-focused preventive intervention designed to reduce children's post-divorce mental health problems, affected attitudes toward divorce and marriage in young adults whose mothers had participated 15 years earlier. Participants (M = 25.6 years; 50% female; 88% Caucasian) were from 240 families that had participated in a randomized experimental trial (NBP vs. literature control). Analyses of covariance showed that program effects on both types of attitudes were moderated by gender. Males in the NBP reported more positive attitudes toward marriage and less favorable attitudes toward divorce than males in the literature control.

DLPFC transcriptome defines two molecular subtypes of schizophrenia.

Little is known about the molecular pathogenesis of schizophrenia, possibly because of unrecognized heterogeneity in diagnosed patient populations. We analyzed gene expression data collected from the dorsolateral prefrontal cortex (DLPFC) of post-mortem frozen brains of 189 adult diagnosed schizophrenics and 206 matched controls. Transcripts from 633 genes are differentially expressed in the DLPFC of schizophrenics as compared to controls at Bonferroni-corrected significance levels. Seventeen of those genes are differentially expressed at very high significance levels (<10-8 after Bonferroni correction). The findings were closely replicated in a dataset from an entirely unrelated source. The statistical significance of this differential gene expression is being driven by about half of the schizophrenic DLPFC samples, and importantly, it is the same half of the samples that is driving the significance for almost all of the differentially expressed transcripts. Weighted gene co-expression network analysis (WGCNA) of the schizophrenic subjects, based on the transcripts differentially expressed in the schizophrenics as compared to controls, divides them into two groups. "Type 1" schizophrenics have a DLPFC transcriptome similar to that of controls with only four differentially expressed genes identified. "Type 2" schizophrenics have a DLPFC transcriptome dramatically different from that of controls, with 3529 expression array probes to 3092 genes detecting transcripts that are differentially expressed at very high significance levels. These findings were re-tested and replicated in a separate independent cohort, using the RNAseq data from the DLPFC of an independent set of schizophrenics and control subjects. We suggest the hypothesis that these striking differences in DLPFC transcriptomes, identified and replicated in two populations, imply a fundamental biologic difference between these two groups of diagnosed schizophrenics, and we propose specific paths for further testing and expanding the hypothesis.

Identification of the Novel Tooth-Specific Transcription Factor AmeloD.

Basic-helix-loop-helix (bHLH) transcription factors play an important role in various organs' development; however, a tooth-specific bHLH factor has not been reported. In this study, we identified a novel tooth-specific bHLH transcription factor, which we named AmeloD, by screening a tooth germ complementary DNA (cDNA) library using a yeast 2-hybrid system. AmeloD was mapped onto the mouse chromosome 1q32. Phylogenetic analysis showed that AmeloD belongs to the achaete-scute complex-like ( ASCL) gene family and is a homologue of ASCL5. AmeloD was uniquely expressed in the inner enamel epithelium (IEE), but its expression was suppressed after IEE cell differentiation into ameloblasts. Furthermore, AmeloD expression showed an inverse expression pattern with the epithelial cell-specific cell-cell adhesion molecule E-cadherin in the dental epithelium. Overexpression of AmeloD in dental epithelial cell line CLDE cells resulted in E-cadherin suppression. We found that AmeloD bound to E-box cis-regulatory elements in the proximal promoter region of the E-cadherin gene. These results reveal that AmeloD functions as a suppressor of E-cadherin transcription in IEE cells. Our study demonstrated that AmeloD is a novel tooth-specific bHLH transcription factor that may regulate tooth development through the suppression of E-cadherin in IEE cells.

Pulmonary arterial hypertension - progress in understanding the disease and prioritizing strategies for drug development.

Pulmonary arterial hypertension (PAH), at one time a largely overlooked disease, is now the subject of intense study in many academic and biotech groups. The availability of new treatments has increased awareness of the condition. This in turn has driven a change in the demographics of PAH, with an increase in the mean age at diagnosis. The diagnosis of PAH in more elderly patients has highlighted the need for careful phenotyping of patients and for further studies to understand how best to manage pulmonary hypertension associated with, for example, left heart disease. The breadth and depth of expertise focused on unravelling the molecular pathology of PAH has yielded novel insights, including the role of growth factors, inflammation and metabolic remodelling. The description of the genetic architecture of PAH is accelerating in parallel, with novel variants, such as those reported in potassium two-pore domain channel subfamily K member 3 (KCNK3), adding to the list of more established mutations in genes associated with bone morphogenetic protein receptor type 2 (BMPR2) signalling. These insights have supported a paradigm shift in treatment strategies away from simply addressing the imbalance of vasoactive mediators observed in PAH towards tackling more directly the structural remodelling of the pulmonary vasculature. Here, we summarize the changing clinical and molecular landscape of PAH. We highlight novel drug therapies that are in various stages of clinical development, targeting for example cell proliferation, metabolic, inflammatory/immune and BMPR2 dysfunction, and the challenges around developing these treatments. We argue that advances in the treatment of PAH will come through deep molecular phenotyping with the integration of clinical, genomic, transcriptomic, proteomic and metabolomic information in large populations of patients through international collaboration. This approach provides the best opportunity for identifying key signalling pathways, both as potential drug targets and as biomarkers for patient selection. The expectation is that together these will enable the prioritization of potential therapies in development and the evolution of personalized medicine for PAH.

Alpha-synuclein (SNCA) polymorphisms exert protective effects on memory after mild traumatic brain injury.

Problems with attention and short-term learning and memory are commonly reported after mild traumatic brain injury (mTBI). Due to the known relationships between α-synuclein (SNCA), dopaminergic transmission, and neurologic deficits, we hypothesized that SNCA polymorphisms might be associated with cognitive outcome after mTBI. A cohort of 91 mTBI patients one month after injury and 86 healthy controls completed a series of cognitive tests assessing baseline intellectual function, attentional function, and memory, and was genotyped at 13 common single nucleotide polymorphisms (SNPs) in the SNCA gene. Significant differences in two memory measures (p=0.001 and 0.002), but not baseline intellectual function or attentional function tasks, were found between the mTBI group and controls. A highly significant protective association between memory performance and SNCA promoter SNP rs1372525 was observed in the mTBI patients (p=0.006 and 0.029 for the long and short delay conditions of the California Verbal Learning Tests, respectively), where the presence of at least one copy of the A (minor) allele was protective after mTBI. These results may help elucidate the pathophysiology of cognitive alterations after mTBI, and thus warrant further investigation.

Regulatory T cells are not a strong predictor of survival for patients with glioblastoma.

BACKGROUND: Regulatory T cells (Tregs) are potentially prognostic indicators in patients with glioblastoma. If differences in frequency of Tregs in tumor or blood account for substantial variation in patient survival, then reliably measuring Tregs may enhance treatment selection and improve outcomes. METHODS: We measured Tregs and CD3+ T cells in tumors and blood from 25 patients with newly diagnosed glioblastoma. Tumor-infiltrating Tregs and CD3+ T cells, measured by quantitative DNA demethylation analysis (epigenetic qPCR) and by immunohistochemistry, and peripheral blood Treg proportions measured by flow cytometry were correlated with patient survival. Additionally, we analyzed data from The Cancer Genome Atlas (TCGA) to correlate the expression of Treg markers with patient survival and glioblastoma subtypes. RESULTS: Tregs, as measured in tumor tissue and peripheral blood, did not correlate with patient survival. Although there was a correlation between tumor-infiltrating Tregs expression by epigenetic qPCR and immunohistochemistry, epigenetic qPCR was more sensitive and specific. Using data from TCGA, mRNA expression of Forkhead box protein 3 (FoxP3) and Helios and FoxP3 methylation level did not predict survival. While the classical glioblastoma subtype corresponded to lower expression of Treg markers, these markers did not predict survival in any of the glioblastoma subtypes. CONCLUSIONS: Although immunosuppression is a hallmark of glioblastoma, Tregs as measured in tissue by gene expression, immunohistochemistry, or demethylation and Tregs in peripheral blood measured by flow cytometry do not predict survival of patients. Quantitative DNA demethylation analysis provides an objective, sensitive, and specific way of identifying Tregs and CD3+ T cells in glioblastoma.

Genetic influences on nicotinic α5 receptor (CHRNA5) CpG methylation and mRNA expression in brain and adipose tissue.

INTRODUCTION: The nicotinic α5 receptor subunit, encoded by CHRNA5, harbors multiple functional single nucleotide polymorphisms (SNPs) that affect mRNA expression and alter the encoded protein. These polymorphisms are most notably associated with drug-taking behaviors and cognition. We previously identified common SNPs in a distant regulatory element (DRE) that increase CHRNA5 mRNA expression in the human prefrontal cortex (PFC) and confer risk for nicotine dependence. Genome-wide epigenetic studies in PFC and adipose tissue find strong effects of the DRE SNPs on CpG methylation. However, it is unclear whether DRE SNPs influence CpG methylation en route to modulating CHRNA5 mRNA expression. It is also unclear whether these polymorphisms affect expression in other brain regions, especially those mediating drug-taking behaviors. RESULTS: By measuring total and allelic CHRNA5 mRNA expression in human habenula and putamen autopsy tissues, we found that CHRNA5 DRE variants considerably increase mRNA expression by up to 3.5-fold in both brain regions. Our epigenetic analysis finds no association between CpG methylation and CHRNA5 mRNA expression in the PFC or adipose tissues. CONCLUSIONS: These finding suggests the mechanisms responsible for the genetic modulation of CpG methylation and mRNA expression are independent despite the DRE SNPs being highly associated with both measures. Our findings support a strong association between the DRE SNPs and mRNA expression or CpG methylation in the brain and periphery, but the independence of the two measures leads us to conclude that environmental factors affecting CpG methylation do not appear to directly modulate gene expression.