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Antipsychotic drugs remain the standard for schizophrenia treatment. Despite their effectiveness in treating hallucinations and delusions, prolonged exposure to antipsychotic medications leads to cognitive deficits in both schizophrenia patients and animal models. The molecular mechanisms underlying these negative effects on cognition remain to be elucidated. Here we demonstrate that chronic antipsychotic drug exposure increases nuclear translocation of NF-κB in both mouse and human frontal cortex, a trafficking event triggered via 5-HT 2A -receptor-dependent downregulation of the NF-κB repressor IκBα. This upregulation of NF-κB activity led to its increased binding at the Hdac2 promoter, thereby augmenting Hdac2 transcription. Deletion of HDAC2 in forebrain pyramidal neurons prevented the negative effects of antipsychotic treatment on synaptic remodeling and cognition. Conversely, virally mediated activation of NF-κB signaling decreased cortical synaptic plasticity via HDAC2. Together, these observations may aid in developing therapeutic strategies to improve the outcome of schizophrenia treatment.

Modulation of transcription, either synthetic activation or repression, via dCas9-fusion proteins is a relatively new methodology with the potential to facilitate high-throughput up- or downregulation studies of gene function. Genetic studies of neurodevelopmental disorders have identified a growing list of risk variants, including both common single-nucleotide variants and rare copy-number variations, many of which are associated with genes having limited functional annotations. By applying a CRISPR-mediated gene-activation/repression platform to populations of human-induced pluripotent stem cell-derived neural progenitor cells, neurons, and astrocytes, we demonstrate that it is possible to manipulate endogenous expression levels of candidate neuropsychiatric risk genes across these three cell types. Although proof-of-concept studies using catalytically inactive Cas9-fusion proteins to modulate transcription have been reported, here we present a detailed survey of the reproducibility of gRNA positional effects across a variety of neurodevelopmental disorder-relevant risk genes, donors, neural cell types, and dCas9 effectors., (Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.)

We report locus-specific disintegration of megabase-scale chromosomal conformations in brain after neuronal ablation of Setdb1 (also known as Kmt1e; encodes a histone H3 lysine 9 methyltransferase), including a large topologically associated 1.2-Mb domain conserved in humans and mice that encompasses >70 genes at the clustered protocadherin locus (hereafter referred to as cPcdh). The cPcdh topologically associated domain (TAD cPcdh ) in neurons from mutant mice showed abnormal accumulation of the transcriptional regulator and three-dimensional (3D) genome organizer CTCF at cryptic binding sites, in conjunction with DNA cytosine hypomethylation, histone hyperacetylation and upregulated expression. Genes encoding stochastically expressed protocadherins were transcribed by increased numbers of cortical neurons, indicating relaxation of single-cell constraint. SETDB1-dependent loop formations bypassed 0.2-1 Mb of linear genome and radiated from the TAD cPcdh fringes toward cis-regulatory sequences within the cPcdh locus, counterbalanced shorter-range facilitative promoter-enhancer contacts and carried loop-bound polymorphisms that were associated with genetic risk for schizophrenia. We show that the SETDB1 repressor complex, which involves multiple KRAB zinc finger proteins, shields neuronal genomes from excess CTCF binding and is critically required for structural maintenance of TAD cPcdh .

Objectives: Despite progress in identifying molecular pathophysiological processes in schizophrenia, valid biomarkers are lacking for both the disease and treatment response., Methods: This comprehensive review summarises recent efforts to identify molecular mechanisms on the level of protein and gene expression and epigenetics, including DNA methylation, histone modifications and micro RNA expression. Furthermore, it summarises recent findings of alterations in lipid mediators and highlights inflammatory processes. The potential that this research will identify biomarkers of schizophrenia is discussed., Results: Recent studies have not identified clear biomarkers for schizophrenia. Although several molecular pathways have emerged as potential candidates for future research, a complete understanding of these metabolic pathways is required to reveal better treatment modalities for this disabling condition., Conclusions: Large longitudinal cohort studies are essential that pair a thorough phenotypic and clinical evaluation for example with gene expression and proteome analysis in blood at multiple time points. This approach might identify biomarkers that allow patients to be stratified according to treatment response and ideally also allow treatment response to be predicted. Improved knowledge of molecular pathways and epigenetic mechanisms, including their potential association with environmental influences, will facilitate the discovery of biomarkers that could ultimately be effective tools in clinical practice.

PHF8 is a histone demethylase with specificity for repressive modifications. While mutations of PHF8 have been associated with cognitive defects and cleft lip/palate, its role in mammalian development and physiology remains unexplored. Here, we have generated a Phf8 knockout allele in mice to examine the consequences of Phf8 loss for development and behaviour. Phf8 deficient mice neither display obvious developmental defects nor signs of cognitive impairment. However, we report a striking resiliency to stress-induced anxiety- and depression-like behaviour on loss of Phf8. We further observe misregulation of serotonin signalling within the prefrontal cortex of Phf8 deficient mice and identify the serotonin receptors Htr1a and Htr2a as direct targets of PHF8. Our results clarify the functional role of Phf8 in mammalian development and behaviour and establish a direct link between Phf8 expression and serotonin signalling, identifying this histone demethylase as a potential target for the treatment of anxiety and depression.

Successful repair of defective composite resin restorations is considered a conservative treatment. The aim of this study was to compare the repair bond strengths of a methacrylate composite (MC) and a silorane composite (SC) repaired after 5 different intervals. Seventy-two test specimens of each material (MC and SC) were prepared. The specimens of each material were divided into 6 groups (n = 12): 1, control (additional composite bonded immediately after polymerization of the substrate composite resin); 2, repaired after 20 minutes; 3, repaired after 24 hours; 4, repaired after 1 week; 5, repaired after 1 month; and 6, repaired after 6 months. Repair consisted of placement of a layer of adhesive bonding agent and then new composite using a plastic mold. Each repair was made with the same material as the original specimen but in a different shade to facilitate fracture assessment. The repaired specimens were stored in distilled water at 37°C for 24 hours before they were submitted to a shear test. There was no statistically significant difference between group MC1 and the other MC groups, except for group MC4, which had lower shear bond strength (SBS) values than groups MC1, MC2, and MC3. Among the SC specimens, all the groups had significantly lower SBS values than group SC1. The mean SBS values of groups SC4, SC5, and SC6 were significantly lower than the SBS of group SC2. All the SC repair time groups presented significantly lower SBS values compared to their corresponding MC groups (P ≤ 0.004). The results showed that the repair bond strength of SC was adversely affected at all time periods, while this effect was not detected for MC groups except for group MC4. The SC material exhibited less repairability than the MC material.

Children with ADHD often show symptoms of oppositional defiant disorders (ODD). We investigated the impact of adjuvant risperidone (RISP) to a standard treatment with methylphenidate (MPH) in children with ADHD and symptoms of ODD. Eighty-four children with ADHD and ODD (age: M=8.55; range: 7.28-9.95 years; 73.8% males) took part in a double-blind, randomized, placebo-controlled, clinical trial lasting eight weeks. Participants were randomly assigned either to the MPH+RISP (1mg/kg/d+0.5mg/d) or to the MPH+PLCO (1mg/kg/d+placebo) condition. Symptoms of ADHD, weight, height, and blood pressure were assessed at baseline, and at weeks 2, 4, 6 and 8. Symptoms of ADHD decreased over time, but more so in the MPH+RISP than in the MPH only condition. In the MPH+RISP condition weight, waist circumference and prolactine levels increased over time. Data suggest that adjuvant RISP improved symptoms in children with ADHD and ODD, but weight gain and higher prolactine levels were also observed, which are two alarming side effects. This may become an issue, once children become adolescents, a period of life in which body shape and body self-image are closely linked to self-confidence and peer acceptance. Health care professionals should carefully balance the short-term and long-term costs and benefits of administration of RISP., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Neuropsychiatric disorders have a complex genetic architecture. Human genetic population-based studies have identified numerous heritable sequence and structural genomic variants associated with susceptibility to neuropsychiatric disease. However, these germline variants do not fully account for disease risk. During brain development, progenitor cells undergo billions of cell divisions to generate the ~80 billion neurons in the brain. The failure to accurately repair DNA damage arising during replication, transcription, and cellular metabolism amid this dramatic cellular expansion can lead to somatic mutations. Somatic mutations that alter subsets of neuronal transcriptomes and proteomes can, in turn, affect cell proliferation and survival and lead to neurodevelopmental disorders. The long life span of individual neurons and the direct relationship between neural circuits and behavior suggest that somatic mutations in small populations of neurons can significantly affect individual neurodevelopment. The Brain Somatic Mosaicism Network has been founded to study somatic mosaicism both in neurotypical human brains and in the context of complex neuropsychiatric disorders., (Copyright © 2017, American Association for the Advancement of Science.)

Background: The nervous system may include more than 100 residue-specific posttranslational modifications of histones forming the nucleosome core that are often regulated in cell-type-specific manner. On a genome-wide scale, some of the histone posttranslational modification landscapes show significant overlap with the genetic risk architecture for several psychiatric disorders, fueling PsychENCODE and other large-scale efforts to comprehensively map neuronal and nonneuronal epigenomes in hundreds of specimens. However, practical guidelines for efficient generation of histone chromatin immunoprecipitation followed by deep sequencing (ChIP-seq) datasets from postmortem brains are needed., Methods: Protocols and quality controls are given for the following: 1) extraction, purification, and NeuN neuronal marker immunotagging of nuclei from adult human cerebral cortex; 2) fluorescence-activated nuclei sorting; 3) preparation of chromatin by micrococcal nuclease digest; 4) ChIP for open chromatin-associated histone methylation and acetylation; and 5) generation and sequencing of ChIP-seq libraries., Results: We present a ChIP-seq pipeline for epigenome mapping in the neuronal and nonneuronal nuclei from the postmortem brain. This includes a stepwise system of quality controls and user-friendly data presentation platforms., Conclusions: Our practical guidelines will be useful for projects aimed at histone posttranslational modification mapping in chromatin extracted from hundreds of postmortem brain samples in cell-type-specific manner., Competing Interests: The authors report no biomedical financial interests or potential conflicts of interest., (Copyright © 2016 Society of Biological Psychiatry. All rights reserved.)

Backgrounds: To assess the role of socio-economic inequality in visual impairment (VI) in Varamin district, Iran., Patients and Methods: Using multistage cluster sampling method, 60 clusters (each with 50 subjects) were recruited and underwent clinical eye examinations. Socio-economic status (SES) was identified based on education, occupation, family assets and housing conditions that were measured at the participants' households using a semi-structured questionnaire and a two-step cluster analysis model. In addition, principal component analysis and the concentration index were used to identify the gap between high and low SES groups., Results: Participants were categorized in high (522, 24.4%), moderate (974, 43.1%) and low (763, 33.7%) socio-economic levels. In these levels, the prevalence of VI was 5.9% (95% CI: 3.3 to 8.6), 10.4% (95% CI: 8.4 to 12.4), and 12.6% (95% CI: 10.1 to 15.1), respectively. The prevalence of VI in people with low SES was significantly greater than those in high SES level. The proportions of avoidable causes were relatively high in all SES levels (more than 80%) with no significant difference between different levels., Conclusions: There is significant inequality in VI prevalence in Varamin district. Avoidable causes are high in all SES groups. Therefore, community-based modalities and preventive programs with a specific notice to poorer SES groups are recommended to improve eye health in this district.

Lysine (K) methyltransferase 2a (Kmt2a) and other regulators of H3 lysine 4 methylation, a histone modification enriched at promoters and enhancers, are widely expressed throughout the brain, but molecular and cellular phenotypes in subcortical areas remain poorly explored. We report that Kmt2a conditional deletion in postnatal forebrain is associated with excessive nocturnal activity and with absent or blunted responses to stimulant and dopaminergic agonist drugs, in conjunction with near-complete loss of spike-timing-dependent long-term potentiation in medium spiny neurons (MSNs). Selective ablation of Kmt2a, but not the ortholog Kmt2b, in adult ventral striatum/nucleus accumbens neurons markedly increased anxiety scores in multiple behavioral paradigms. Striatal transcriptome sequencing in adult mutants identified 262 Kmt2a-sensitive genes, mostly downregulated in Kmt2a-deficient mice. Transcriptional repression includes the 5-Htr2a serotonin receptor, strongly associated with anxiety- and depression-related disorders in human and animal models. Consistent with the role of Kmt2a in promoting gene expression, the transcriptional regulators Bahcc1, Isl1, and Sp9 were downregulated and affected by H3K4 promoter hypomethylation. Therefore, Kmt2a regulates synaptic plasticity in striatal neurons and provides an epigenetic drug target for anxiety and dopamine-mediated behaviors.

Background: Early childhood malnutrition affects 113 million children worldwide, impacting health and increasing vulnerability for cognitive and behavioral disorders later in life. Molecular signatures after childhood malnutrition, including the potential for intergenerational transmission, remain unexplored., Methods: We surveyed blood DNA methylomes (~483,000 individual CpG sites) in 168 subjects across two generations, including 50 generation 1 individuals hospitalized during the first year of life for moderate to severe protein-energy malnutrition, then followed up to 48 years in the Barbados Nutrition Study. Attention deficits and cognitive performance were evaluated with the Connors Adult Attention Rating Scale and Wechsler Abbreviated Scale of Intelligence. Expression of nutrition-sensitive genes was explored by quantitative reverse transcriptase polymerase chain reaction in rat prefrontal cortex., Results: We identified 134 nutrition-sensitive, differentially methylated genomic regions, with most (87%) specific for generation 1. Multiple neuropsychiatric risk genes, including COMT, IFNG, MIR200B, SYNGAP1, and VIPR2 showed associations of specific methyl-CpGs with attention and IQ. IFNG expression was decreased in prefrontal cortex of rats showing attention deficits after developmental malnutrition., Conclusions: Early childhood malnutrition entails long-lasting epigenetic signatures associated with liability for attention and cognition, and limited potential for intergenerational transmission., Competing Interests: Competing Financial Interests: The authors report no biomedical financial interests or potential conflicts of interest., (Copyright © 2016 Society of Biological Psychiatry. Published by Elsevier Inc. All rights reserved.)

The Psychiatric Genomics Consortium-Schizophrenia Workgroup (PGC-SCZ) recently identified 108 loci associated with increased risk for schizophrenia (SCZ). The vast majority of these variants reside within non-coding sequences of the genome and are predicted to exert their effects by affecting the mechanism of action of cis regulatory elements (CREs), such as promoters and enhancers. Although a number of large-scale collaborative efforts (e.g. ENCODE) have achieved a comprehensive mapping of CREs in human cell lines or tissue homogenates, it is becoming increasingly evident that many risk-associated variants are enriched for expression Quantitative Trait Loci (eQTLs) and CREs in specific tissues or cells. As such, data derived from previous research endeavors may not capture fully cell-type and/or region specific changes associated with brain diseases. Coupling recent technological advances in genomics with cell-type specific methodologies, we are presented with an unprecedented opportunity to better understand the genetics of normal brain development and function and, in turn, the molecular basis of neuropsychiatric disorders. In this review, we will outline ongoing efforts towards this goal and will discuss approaches with the potential to shed light on the mechanism(s) of action of cell-type specific cis regulatory elements and their putative roles in disease, with particular emphasis on understanding the manner in which the epigenome and CREs influence the etiology of SCZ., Competing Interests: The authors whose names are listed immediately below certify that they have NO affiliations with or involvement in any organization or entity with any financial interest (such as honoraria; educational grants; participation in speakers’ bureaus; membership, employment, consultancies, stock ownership, or other equity interest; and expert testimony or patent-licensing arrangements), or non-financial interest (such as personal or professional relationships, affiliations, knowledge or beliefs) in the subject matter or materials discussed in this manuscript., (Published by Elsevier B.V.)

Nonrandom chromosomal conformations, including promoter-enhancer loopings that bypass kilobases or megabases of linear genome, provide a crucial layer of transcriptional regulation and move vast amounts of non-coding sequence into the physical proximity of genes that are important for neurodevelopment, cognition and behaviour. Activity-regulated changes in the neuronal '3D genome' could govern transcriptional mechanisms associated with learning and plasticity, and loop-bound intergenic and intronic non-coding sequences have been implicated in psychiatric and adult-onset neurodegenerative disease. Recent studies have begun to clarify the roles of spatial genome organization in normal and abnormal cognition.

Psychiatric disorders are complex multifactorial illnesses involving chronic alterations in neural circuit structure and function as well as likely abnormalities in glial cells. While genetic factors are important in the etiology of most mental disorders, the relatively high rates of discordance among identical twins, particularly for depression and other stress-related syndromes, clearly indicate the importance of additional mechanisms. Environmental factors such as stress are known to play a role in the onset of these illnesses. Exposure to such environmental insults induces stable changes in gene expression, neural circuit function, and ultimately behavior, and these maladaptations appear distinct between developmental versus adult exposures. Increasing evidence indicates that these sustained abnormalities are maintained by epigenetic modifications in specific brain regions. Indeed, transcriptional dysregulation and the aberrant epigenetic regulation that underlies this dysregulation is a unifying theme in psychiatric disorders. Here, we provide a progress report of epigenetic studies of the three major psychiatric syndromes, depression, schizophrenia, and bipolar disorder. We review the literature derived from animal models of these disorders as well as from studies of postmortem brain tissue from human patients. While epigenetic studies of mental illness remain at early stages, understanding how environmental factors recruit the epigenetic machinery within specific brain regions to cause lasting changes in disease susceptibility and pathophysiology is revealing new insight into the etiology and treatment of these conditions., Competing Interests: Declaration of Conflicting Interests The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2015.)

Cognitive defects in autism spectrum disorder (ASD) include socialization and communication: key behavioral capacities that separate humans from other species. Here, we analyze gene expression in the prefrontal cortex of 63 autism patients and control individuals, as well as 62 chimpanzees and macaques, from natal to adult age. We show that among all aberrant expression changes seen in ASD brains, a single aberrant expression pattern overrepresented in genes involved synaptic-related pathways is enriched in nucleotide variants linked to autism. Furthermore, only this pattern contains an excess of developmental expression features unique to humans, thus resulting in the disruption of human-specific developmental programs in autism. Several members of the early growth response (EGR) transcription factor family can be implicated in regulation of this aberrant developmental change. Our study draws a connection between the genetic risk architecture of autism and molecular features of cortical development unique to humans., Competing Interests: The authors have declared that no competing interests exist.

Neuronal epigenomes, including chromosomal loopings moving distal cis-regulatory elements into proximity of target genes, could serve as molecular proxy linking present-day-behaviour to past exposures. However, longitudinal assessment of chromatin state is challenging, because conventional chromosome conformation capture assays essentially provide single snapshots at a given time point, thus reflecting genome organization at the time of brain harvest and therefore are non-informative about the past. Here we introduce 'NeuroDam' to assess epigenome status retrospectively. Short-term expression of the bacterial DNA adenine methyltransferase Dam, tethered to the Gad1 gene promoter in mouse prefrontal cortex neurons, results in stable G(methyl)ATC tags at Gad1-bound chromosomal contacts. We show by NeuroDam that mice with defective cognition 4 months after pharmacological NMDA receptor blockade already were affected by disrupted chromosomal conformations shortly after drug exposure. Retrospective profiling of neuronal epigenomes is likely to illuminate epigenetic determinants of normal and diseased brain development in longitudinal context.

Objective: To evaluate shear bond strength (SBS) of the orthodontic brackets bonded to fluorosed and nonfluorosed teeth using Light Bond with and without adhesion promoters and compare their enamel damages following debonding., Materials and Methods: In this study, 30 fluorosed (Thylstrup and Fejerskov Index = 4-5) and 30 nonfluorosed teeth were randomly distributed between two subgroups according to the bonding materials: Group 1, fluorosed teeth bonded with Light Bond; Group 2, fluorosed teeth bonded with adhesion promoters and Light Bond; Group 3, nonfluorosed teeth bonded with Light Bond; Group 4, nonfluorosed bonded with adhesion promoters and Light Bond. After bonding, the SBS of the brackets was tested with a universal testing machine. Stereomicroscopic evaluation was performed by unbiased stereology in all teeth to determine the amount of adhesive remnants and the number and length of enamel cracks before bonding and after debonding. The data were analyzed using two-way analysis of variance, Kruskal-Wallis, Wilcoxon Signed Rank, and Mann-Whitney test., Results: While fluorosis reduced the SBS of orthodontic bracket (P = 0.017), Enhance Locus Ceruleus LC significantly increased the SBS of the orthodontic bracket in fluorosed and nonfluorosed teeth (P = 0.039). Significant increasing in the number and length of enamel crack after debonding was found in all four groups. There were no significant differences in the length of enamel crack increased after debonding among four groups (P = 0.768) while increasing in the number of enamel cracks after debonding was significantly different among the four groups (P = 0.023). Teeth in Group 2 showed the highest enamel damages among four groups following debonding., Conclusion: Adhesion promoters could improve the bond strength of orthodontic brackets, but conservative debonding methods for decreasing enamel damages would be necessary.

Attention is disrupted commonly in psychiatric disorders, yet mechanistic insight remains limited. Deficits in this function are associated with dorsal anterior cingulate cortex (dACC) excitotoxic lesions and pharmacological disinhibition; however, a causal relationship has not been established at the cellular level. Moreover, this association has not yet been examined in a genetically tractable species such as mice. Here, we reveal that dACC neurons causally contribute to attention processing by combining a chemogenetic approach that reversibly suppresses neural activity with a translational, touchscreen-based attention task in mice. We virally expressed inhibitory hM4Di DREADD (designer receptor exclusively activated by a designer drug) in dACC neurons, and examined the effects of this inhibitory action with the attention-based five-choice serial reaction time task. DREADD inactivation of the dACC neurons during the task significantly increased omission and correct response latencies, indicating that the neuronal activities of dACC contribute to attention and processing speed. Selective inactivation of excitatory neurons in the dACC not only increased omission, but also decreased accuracy. The effect of inactivating dACC neurons was selective to attention as response control, motivation, and locomotion remain normal. This finding suggests that dACC excitatory neurons play a principal role in modulating attention to task-relevant stimuli. This study establishes a foundation to chemogenetically dissect specific cell-type and circuit mechanisms underlying attentional behaviors in a genetically tractable species.

Increased neuronal densities in subcortical white matter have been reported for some cases with schizophrenia. The underlying cellular and molecular mechanisms remain unresolved. We exposed 26 young adult macaque monkeys for 6 months to either clozapine, haloperidol or placebo and measured by structural MRI frontal gray and white matter volumes before and after treatment, followed by observer-independent, flow-cytometry-based quantification of neuronal and non-neuronal nuclei and molecular fingerprinting of cell-type specific transcripts. After clozapine exposure, the proportion of nuclei expressing the neuronal marker NeuN increased by approximately 50% in subcortical white matter, in conjunction with a more subtle and non-significant increase in overlying gray matter. Numbers and proportions of nuclei expressing the oligodendrocyte lineage marker, OLIG2, and cell-type specific RNA expression patterns, were maintained after antipsychotic drug exposure. Frontal lobe gray and white matter volumes remained indistinguishable between antipsychotic-drug-exposed and control groups. Chronic clozapine exposure increases the proportion of NeuN+ nuclei in frontal subcortical white matter, without alterations in frontal lobe volumes or cell type-specific gene expression. Further exploration of neurochemical plasticity in non-human primate brain exposed to antipsychotic drugs is warranted., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Astrocytes orchestrate arrangement and functions of neuronal circuits and of the blood-brain barrier. Dysfunctional astrocytes characterize mood disorders, here showcased by deregulation of the astrocyte end-feet protein Aquaporin-4 around blood vessels and, hypothetically, of the astrocyte-specific phagocytic protein MEGF10 to shape synapses. Development of mood disorders is often a result of 'gene × environment' interactions, regulated among others by histone modifications and related modulator enzymes, which rapidly promote adaptive responses. Thus, they represent ideal targets of drugs aimed at inducing stable effects with quick onsets. One of the prevalent features of histone modifications and their modulators is their cell-type specificity. Investigating cell type-specific epigenetic modulations upon drug administration might therefore help to implement therapeutic treatments., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Huntington's disease (HD) is an autosomal-dominant neurodegenerative disorder resulting from expansion of CAG repeats in the Huntingtin (HTT) gene. Previous studies have shown mutant HTT can alter expression of genes associated with dysregulated epigenetic modifications. One of the most widely studied chromatin modifications is trimethylated lysine 4 of histone 3 (H3K4me3). Here, we conducted the first comprehensive study of H3K4me3 ChIP-sequencing in neuronal chromatin from the prefrontal cortex of six HD cases and six non-neurologic controls, and its association with gene expression measured by RNA-sequencing. We detected 2,830 differentially enriched H3K4me3 peaks between HD and controls, with 55% of them down-regulated in HD. Although H3K4me3 signals are expected to be associated with mRNA levels, we found an unexpected discordance between altered H3K4me3 peaks and mRNA levels. Gene ontology (GO) term enrichment analysis of the genes with differential H3K4me3 peaks, revealed statistically significantly enriched GO terms only in the genes with down-regulated signals in HD. The most frequently implicated biological process terms are organ morphogenesis and positive regulation of gene expression. More than 9,000 H3K4me3 peaks were located not near any recognized transcription start sites and approximately 36% of these "distal" peaks co-localized to known enhancer sites. Six transcription factors and chromatin remodelers are differentially enriched in HD H3K4me3 distal peaks, including EZH2 and SUZ12, two core subunits of the polycomb repressive complex 2 (PRC2). Moreover, PRC2 repressive state was significantly depleted in HD-enriched peaks, suggesting the epigenetic role of PRC2 inhibition associated with up-regulated H3K4me3 in Huntington's disease. In summary, our study provides new insights into transcriptional dysregulation of Huntington's disease by analyzing the differentiation of H3K4me3 enrichment.

Huntington's Disease (HD) is a devastating neurodegenerative disorder that is caused by an expanded CAG trinucleotide repeat in the Huntingtin (HTT) gene. Transcriptional dysregulation in the human HD brain has been documented but is incompletely understood. Here we present a genome-wide analysis of mRNA expression in human prefrontal cortex from 20 HD and 49 neuropathologically normal controls using next generation high-throughput sequencing. Surprisingly, 19% (5,480) of the 28,087 confidently detected genes are differentially expressed (FDR<0.05) and are predominantly up-regulated. A novel hypothesis-free geneset enrichment method that dissects large gene lists into functionally and transcriptionally related groups discovers that the differentially expressed genes are enriched for immune response, neuroinflammation, and developmental genes. Markers for all major brain cell types are observed, suggesting that HD invokes a systemic response in the brain area studied. Unexpectedly, the most strongly differentially expressed genes are a homeotic gene set (represented by Hox and other homeobox genes), that are almost exclusively expressed in HD, a profile not widely implicated in HD pathogenesis. The significance of transcriptional changes of developmental processes in the HD brain is poorly understood and warrants further investigation. The role of inflammation and the significance of non-neuronal involvement in HD pathogenesis suggest anti-inflammatory therapeutics may offer important opportunities in treating HD.

DNA methylation patterns change with age, and aging itself is a major confounding risk factor for Parkinson's disease (PD). Duplication and triplication, that is, increased expression of the α-synuclein (SNCA) gene, cause familial PD, and demethylation of SNCA intron 1 has been shown to result in increased expression of SNCA. We thus hypothesized that age-related alterations of SNCA methylation might underly the increased susceptibility toward PD in later life. The present study sought to determine (1) whether alterations of SNCA intron 1 methylation occurred during aging, (2) whether the methylation pattern differed between men and women, and (3) whether purified neurons compared with non-neuronal cells exhibited different methylation patterns. The analysis of DNA from brain tissue and fluorescence activated cell sorting-sorted purified neurons of 41 individuals revealed only a minor increase of SNCA intron 1 DNA methylation levels in presumably healthy individuals during aging but no significant difference between men and women. Interestingly enough, methylation of SNCA intron 1 was higher in neurons compared with non-neuronal cells, although non-neuronal cells express lower levels of SNCA. Therefore, the normal pattern of SNCA methylation during aging should not result in increased expression of α-synuclein protein. It is thus likely that additional, yet not identified, mechanisms contribute to the tissue specificity of SNCA expression and the presumed dysregulation in PD., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Social cognition is a complex process that requires the integration of a wide variety of behaviors, including salience, reward-seeking, motivation, knowledge of self and others, and flexibly adjusting behavior in social groups. Not surprisingly, social cognition represents a sensitive domain commonly disrupted in the pathology of a variety of psychiatric disorders including Autism Spectrum Disorder (ASD) and Schizophrenia (SCZ). Here, we discuss convergent research from animal models to human disease that implicates the prefrontal cortex (PFC) as a key regulator in social cognition, suggesting that disruptions in prefrontal microcircuitry play an essential role in the pathophysiology of psychiatric disorders with shared social deficits. We take a translational perspective of social cognition, and review three key behaviors that are essential to normal social processing in rodents and humans, including social motivation, social recognition, and dominance hierarchy. A shared prefrontal circuitry may underlie these behaviors. Social cognition deficits in animal models of neurodevelopmental disorders like ASD and SCZ have been linked to an altered balance of excitation and inhibition (E/I ratio) within the cortex generally, and PFC specifically. A clear picture of the mechanisms by which altered E/I ratio in the PFC might lead to disruptions of social cognition across a variety of behaviors is not well understood. Future studies should explore how disrupted developmental trajectory of prefrontal microcircuitry could lead to altered E/I balance and subsequent deficits in the social domain.

Purpose: To determine the prevalence of amblyopia and refractive errors among 7 to 12-year-old primary school children in Tehran, Iran., Methods: This population-based cross-sectional study included 2,410 randomly selected students. Visual acuity was tested using an E-chart on Yang vision tester. Refractive errors were measured by photorefractometry and cycloautorefraction. Strabismus was checked using cover test. Direct ophthalmoscopy was used to assess the anterior segment, lens opacities, red reflex and fundus. Functional amblyopia was defined as best corrected visual acuity ≤20/40 in one or both eyes with no anatomical problems., Results: Amblyopia was present in 2.3% (95% CI: 1.8% to 2.9%) of participants with no difference between the genders. Amblyopic subjects were significantly younger than non-amblyopic children (P=0.004). Overall, 15.9% of hyperopic and 5.9% of myopic cases had amblyopia. The prevalence of hyperopia ≥+2.00D, myopia ≤-0.50D, astigmatism ≥0.75D, and anisometropia (≥1.00D) was 3.5%, 4.9%, 22.6%, and 3.9%, respectively. With increasing age, the prevalence of myopia increased (P<0.001), that of hyperopia decreased (P=0.007), but astigmatism showed no change. Strabismus was found in 2.3% of cases. Strabismus (OR=17.9) and refractive errors, especially anisometropia (OR=12.87) and hyperopia (OR=11.87), were important amblyogenic risk factors., Conclusion: The high prevalence of amblyopia in our subjects in comparison to developed countries reveals the necessity of timely and sensitive screening methods. Due to the high prevalence of amblyopia among children with refractive errors, particularly high hyperopia and anisometropia, provision of glasses should be specifically attended by parents and supported by the Ministry of Health and insurance organizations.

Schizophrenia, a major psychiatric disorder defined by delusions and hallucinations, among other symptoms, often with onset in early adulthood, is potentially associated with molecular and cellular alterations in parvalbumin-expressing fast spiking interneurons and other constituents of the cortical inhibitory GABAergic circuitry. The underlying mechanisms, including the role of disease-associated risk factors operating in adolescence such as drug abuse and social stressors, remain incompletely understood. Here, we summarize emerging findings from animal models, highlighting the ability of parvalbuminergic interneurons (PVI) to induce, during the juvenile period, long-term plastic changes in prefrontal and visual cortex, thereby altering perception, cognition and behavior in the adult. Of note, molecular alterations in PVI from subjects with schizophrenia, including downregulated expression of a subset of GABAergic genes, have also been found in juvenile stress models of the disorder. Some of the transcriptional alterations observed in schizophrenia postmortem brain could be linked to changes in the epigenetic architecture of GABAergic gene promoters, including dysregulated DNA methylation, histone modification patterns and disruption of promoter-enhancer interactions at site of chromosomal loop formations. Therefore, we predict that, in the not-to-distant future, PVI- and other cell-type specific epigenomic mappings in the animal model and human brain will provide novel insights into the pathophysiology of schizophrenia and related psychotic diseases, including the role of cortical GABAergic circuitry in shaping long-term plasticity and cognitive function of the cerebral cortex., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Expression of GAD1 GABA synthesis enzyme is highly regulated by neuronal activity and reaches mature levels in the prefrontal cortex not before adolescence. A significant portion of cases diagnosed with schizophrenia show deficits in GAD1 RNA and protein levels in multiple areas of adult cerebral cortex, possibly reflecting molecular or cellular defects in subtypes of GABAergic interneurons essential for network synchronization and cognition. Here, we review 20years of progress towards a better understanding of disease-related regulation of GAD1 gene expression. For example, deficits in cortical GAD1 RNA in some cases of schizophrenia are associated with changes in the epigenetic architecture of the promoter, affecting DNA methylation patterns and nucleosomal histone modifications. These localized chromatin defects at the 5' end of GAD1 are superimposed by disordered locus-specific chromosomal conformations, including weakening of long-range promoter-enhancer loopings and physical disconnection of GAD1 core promoter sequences from cis-regulatory elements positioned 50 kilobases further upstream. Studies on the 3-dimensional architecture of the GAD1 locus in neurons, including developmentally regulated higher order chromatin compromised by the disease process, together with exploration of locus-specific epigenetic interventions in animal models, could pave the way for future treatments of psychosis and schizophrenia., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Objectives: Repairing composite restorations is a challenging procedure especially when two different types of composites are used. This study aimed to compare the repair strength of silorane-based composite (SC) (Filtek P90) with that of preheated SC, methacrylate composite (MC)(Z250), flowable MC (Filtek Supreme Plus) and different adhesive/composite combinations., Materials and Methods: Eighty-four SC specimens were fabricated and randomly divided into seven groups (G). In the control group (G7), SC was bonded immediately to SC. The other specimens were water-aged for two months and were then roughened, etched and repaired with the following materials: G1) Silorane Adhesive Bond (SAB)/SC; G2) Preheated SC; G3) SAB/MC; G4) Adper Single Bond (SB)/MC; G5) Flowable MC/MC; G6) Preheated MC. After water storage and thermocycling, the repaired specimens were subjected to shear bond strength testing. The data were analyzed using ANOVA and Tukey's test., Results: Preheated SC and MC, flowable MC and SAB/SC resulted in bond strength comparable to that of the control group. Preheated SC showed significantly higher bond strength when compared to SAB/MC (P=0.04) and SB/MC (P<0.001). Bond strength of SB/MC was significantly lower than that of the other groups (P<0.05), except for SAB/SC and SAB/MC., Conclusion: All repairing materials except for SB/MC resulted in bond strength values comparable to that of the control group. Repair with preheated SC yielded the highest bond strength.

Purpose: To evaluate intrasubject asymmetry in central corneal thickness (CCT) among patients with primary open-angle glaucoma (POAG) as compared with that of normal subjects and to determine whether the eye with thinner cornea has more severe glaucomatous visual field damage., Methods: In this case-control study, CCT of both eyes was measured using an ultrasonic pachymeter (UP-1000; Nidek Technologies, Gamagori, Japan) among POAG patients and normal subjects. The intrasubject difference in CCT was calculated and compared between the study groups; correlations between CCT and intraocular pressure (IOP), vertical cup-to-disc ratio (VCDR), and visual field defects were also evaluated., Results: A total of 62 patients with bilateral POAG and 56 normal subjects were included. There was no significant difference between the study groups in terms of age, sex, and ocular parameters except for visual acuity, IOP, and VCDR. The POAG patients demonstrated significantly greater intrasubject asymmetry in CCT (8 ± 7 μm vs. 5 ± 3 μm; p = 0.041) and a higher prevalence of significant (>10 μm) intrasubject CCT difference (30.6 vs. 5.4%; p < 0.001) as compared with normal subjects. Although each higher level of glaucomatous damage was associated with 4-μm thinner CCT, the correlation was not statistically significant (95% confidence interval, -8 to 1 μm; p = 0.117; GEE analysis). No significant correlation was observed between CCT and IOP or VCDR (p = 0.302 and 0.137, respectively)., Conclusions: Patients with POAG demonstrate a larger amount of intrasubject difference in CCT as compared with normal subjects.

Turnover and exchange of nucleosomal histones and their variants, a process long believed to be static in post-replicative cells, remains largely unexplored in brain. Here, we describe a novel mechanistic role for HIRA (histone cell cycle regulator) and proteasomal degradation-associated histone dynamics in the regulation of activity-dependent transcription, synaptic connectivity, and behavior. We uncover a dramatic developmental profile of nucleosome occupancy across the lifespan of both rodents and humans, with the histone variant H3.3 accumulating to near-saturating levels throughout the neuronal genome by mid-adolescence. Despite such accumulation, H3.3-containing nucleosomes remain highly dynamic-in a modification-independent manner-to control neuronal- and glial-specific gene expression patterns throughout life. Manipulating H3.3 dynamics in both embryonic and adult neurons confirmed its essential role in neuronal plasticity and cognition. Our findings establish histone turnover as a critical and previously undocumented regulator of cell type-specific transcription and plasticity in mammalian brain., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Background: Gamma radiation effects on the erythrocyte membrane from three different functional parts, lipid bilayer, cytoskeleton and protein components. When the red cell membrane is exposed to radiation, it loses its integrity and hemoglobin leaks out. In addition, irradiation leads to lipid peroxidation and the products of this process, leading to hemolysis. The aim of the present study was to measure osmotic fragility (OF) of red blood cells and malondialdehyde (MDA) levels as a marker of oxidative injury in breast cancer patients treated with radiation and chemotherapy., Materials and Methods: The OF test was performed using different concentrations of a salt solution. The measurement of MDA was done with chemical methods.(11) The sampling was taken during three stages of treatment: first sample was taken before starting chemotherapy, the second sample was taken before radiation therapy and the third sample was taken after radiotherapy., Results: No statistically significant differences between levels of MDA in these three stages of treatment were observed. However, the comparison of mean levels of MDA showed an increase after radiotherapy. The OF rate did not show significant difference (P > 0.05) during the stages of treatment., Conclusion: In a standard treatment program of radiotherapy and chemotherapy lipid peroxidation level and OF do not significantly increase.

Background and aims. Nano-ionomer (NI) interacts with tooth structures superficially, and there is a concern about the enamel bonding ability of mild self-etch Ketac primer. This study compared the effect of different adhesive procedures (self-etching and etch-and-rinse approach) on long-term marginal microleakage of nano-filled resin-modified glass-ionomer (NI) cervical restorations. Materials and methods. Class V cavities were prepared on 72 maxillary premolars. The teeth were divided into six groups: G1: No treatment (NC); G2: Ketac primer (K primer); G3: Etchant + Ketac primer (E+K primer); G4: Self-etch adhesive (Bond Force); G5: Etchant + Bond Force (E+Bond Force); G6: Etchant + Adper Single Bond (Etch and rinse adhesive). All the cavities were restored with Ketac N100. The samples were stored in water for 6 months and thermocycled for 2000 cycles. Marginal sealing was assessed using dye penetration technique. Data were analyzed with non-parametric tests (α=0.05). Results. All the adhesive pretreatments resulted in a lower marginal leakage than that of NC (P≤0.01), except for E+Bond Force at the dentin margin. There was no significant difference between K primer and Bond Force. Microleakage differed significantly between K primer pretreatment and E+K primer (P=0.003), E+Bond Force (P=0.002) and etch-and-rinse adhesive (P=0.001) at the enamel margin, but it did not differ at the dentin margin. E+ Bond Force group showed insignificantly lower leakage at the enamel margin and significantly higher leakage at the dentin margin (P=0.02). Conclusion. Etch-and-rinse adhesive and selective enamel etching along with self-etch adhesive/Ketac primer might improve marginal sealing of aged nano-ionomer restoration.

Many neurological and psychiatric disorders exhibit gender disparities, and sex differences in the brain likely explain some of these effects. Recent work in rodents points to a role for epigenetics in the development or maintenance of neural sex differences, although genome-wide studies have so far been lacking. Here we review the existing literature on epigenetics and brain sexual differentiation and present preliminary analyses on the genome-wide distribution of histone-3 lysine-4 trimethylation in a sexually dimorphic brain region in male and female mice. H3K4me3 is a histone mark primarily organized as 'peaks' surrounding the transcription start site of active genes. We microdissected the bed nucleus of the stria terminalis and preoptic area (BNST/POA) in adult male and female mice and used ChIP-Seq to compare the distribution of H3K4me3 throughout the genome. We found 248 genes and loci with a significant sex difference in H3K4me3. Of these, the majority (71%) had larger H3K4me3 peaks in females. Comparisons with existing databases indicate that genes and loci with increased H3K4me3 in females are associated with synaptic function and with expression atlases from related brain areas. Based on RT-PCR, only a minority of genes with a sex difference in H3K4me3 has detectable sex differences in expression at baseline conditions. Together with previous findings, our data suggest that there may be sex biases in the use of epigenetic marks. Such biases could underlie sex differences in vulnerabilities to drugs or diseases that disrupt specific epigenetic processes., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Epigenetic modifications are alterations that regulate gene expression without modifying the underlying DNA sequence. DNA methylation and histone modifications, for example, are capable of spatial and temporal regulation of expression-with several studies demonstrating that these epigenetic marks are heritable. Thus, like DNA sequence, epigenetic marks are capable of storing information and passing it from one generation to the next. Because the epigenome is dynamic and epigenetic modifications can respond to external environmental stimuli, such changes may play an important role in adaptive evolution. While recent studies provide strong evidence for species-specific signatures of epigenetic marks, little is known about the mechanisms by which such modifications evolve. In order to address this question, we analyze the genome wide distribution of an epigenetic histone mark (H3K4me3) in prefrontal cortex neurons of humans, chimps and rhesus macaques. We develop a novel statistical framework to quantify within- and between-species variation in histone methylation patterns, using an ANOVA-based method and defining an FST -like measure for epigenetics (termed epi- FST), in order to develop a deeper understanding of the evolutionary pressures acting on epigenetic variation. Results demonstrate that genes with high epigenetic FST values are indeed significantly overrepresented among genes that are differentially expressed between species, and we observe only a weak correlation with SNP density.

Alterations in histone lysine methylation and epigenetic regulators of gene expression could play a role in the neurobiology and treatment of patients diagnosed with mood spectrum disorder, including depression and anxiety. Mutations and altered expression of various lysine methyltransferases (KMTs) and demethylases (KDMs) have been linked to changes in motivational and emotional behaviors in preclinical model systems. However, it is not known whether regulators operating downstream of histone lysine methylation could affect mood-related behavior. Malignant Brain Tumor (MBT) domain 'chromatin reader' proteins bind to methylated histone lysine residues and associate with chromatin remodeling complexes to facilitate or repress gene expression. MBT proteins, including the founding member, L3mbtl1, maintain high levels of expression in neurons of the mature brain. Here, we exposed L3mbtl1 null mutant mice to a wide range of tests exploring cognition and mood-relevant behaviors at baseline and in the context of social isolation, as a stressor to elicit depression-related behavior in susceptible mice. L3mbtl1 loss-of-function was associated with significant decreases in depression and and anxiety in some of the behavioral paradigms. This was not associated with a more generalized neurological dysfunction because cognition and memory remained unaltered in comparison to controls. These findings warrant further investigations on the role of MBT chromatin reader proteins in the context of emotional and affective behaviors.

Background: A recent survey of avoidable blindness in Varamin District, Iran, identified moderately high levels of visual impairment (10%) and blindness (1.5%) in people >50 years. This study aimed to define current provision, identify gaps and suggest practical solutions for improving eye health services in this area., Methods: The World Health Organization (WHO) framework for analyzing health systems has several key components: service delivery, health workforce, information system, medical products and technologies, financing, and governance. We used this structure to investigate the strengths and weaknesses of the eye health system in Varamin. All public and private eye care facilities and a random selection of primary health care (PHC) units were assessed using semi-structured researcher-administered questionnaires., Results: Varamin has 16 ophthalmic clinics, including two secondary hospitals that provide cataract surgery. There were ten ophthalmologists (1:68,000 population), two ophthalmic nurses and five optometrists working in Varamin district. There were no eye care social or community workers, ophthalmic counsellors, low vision rehabilitation staff. Although the Vision 2020 target for ophthalmologists has been met, numbers of other eye care staff were insufficient. The majority of patients travel to Tehran for surgery. The recent survey identified cataract as the leading cause of blindness, despite the availability of surgical services in the district and high health insurance coverage. Poor awareness is a major barrier. No units had a written blindness prevention plan, formal referral pathways or sufficient eye health promotion activities. Only one of the PHC units referred people with diabetes for retinal examination. There is partial integration between eye care services and the general health system particularly for prevention of childhood blindness: chemo-prophylaxis for ophthalmia neonatorum, school vision tests, measles immunization and Vitamin A supplementation., Conclusions: This analysis demonstrated the need for better integration between eye care services and the general health system, local planning for prevention of blindness, an information system, a better staff mix and health education to increase community awareness and service uptake. There is the capacity to deliver far more surgery locally. All aspects of a health system need to be developed to deliver comprehensive and efficient eye care.

Purpose: To assess the association of LTBP2 mutations with primary angle closure glaucoma (PACG)., Methods: We studied 54 unrelated patients with PACG and one individual with pseudoexfoliation accompanied with angle closure glaucoma; these consisted of 28 female and 27 male subjects aged 27 to 82 (mean, 63) years. The 36 exons and flanking intronic sequences of LTBP2 in all patients were amplified by PCR and sequenced by the Sanger protocol. The sequences were compared to LTBP2 reference sequences. A total of 100 to 400 controls aged at least 60 years old were screened for various variations., Results: Out of 24 observed sequence variations, ten were in amino acid coding regions; of these four created synonymous codons while six caused amino acid changes. Based on allele frequencies, biochemical parameters, absence in control individuals, evolutionary conservation of affected amino acids, and bioinformatic predictions on the effects on protein function, it was concluded that only two mutations causing p. Gln1417Arg and p. Gly1660Trp may contribute to PACG. The p. Gly1660Trp mutation was observed in a patient with both PACG and PEX syndrome. P. Gln1417Arg had previously been reported only in a subject with POAG., Conclusion: LTBP2 may contribute to PACG. This finding emphasizes that there may be an overlap in the etiology of various forms of glaucoma and the overlaps likely contribute to common features in various forms of glaucoma.

Background: MicroRNAs (miRNAs) are small non-coding RNAs that recognize sites of complementarity of target messenger RNAs, resulting in transcriptional regulation and translational repression of target genes. In Huntington's disease (HD), a neurodegenerative disease caused by a trinucleotide repeat expansion, miRNA dyregulation has been reported, which may impact gene expression and modify the progression and severity of HD., Methods: We performed next-generation miRNA sequence analysis in prefrontal cortex (Brodmann Area 9) from 26 HD, 2 HD gene positive, and 36 control brains. Neuropathological information was available for all HD brains, including age at disease onset, CAG-repeat size, Vonsattel grade, and Hadzi-Vonsattel striatal and cortical scores, a continuous measure of the extent of neurodegeneration. Linear models were performed to examine the relationship of miRNA expression to these clinical features, and messenger RNA targets of associated miRNAs were tested for gene ontology term enrichment., Results: We identified 75 miRNAs differentially expressed in HD brain (FDR q-value <0.05). Among the HD brains, nine miRNAs were significantly associated with Vonsattel grade of neuropathological involvement and three of these, miR-10b-5p, miR-10b-3p, and miR-302a-3p, significantly related to the Hadzi-Vonsattel striatal score (a continuous measure of striatal involvement) after adjustment for CAG length. Five miRNAs (miR-10b-5p, miR-196a-5p, miR-196b-5p, miR-10b-3p, and miR-106a-5p) were identified as having a significant relationship to CAG length-adjusted age of onset including miR-10b-5p, the mostly strongly over-expressed miRNA in HD cases. Although prefrontal cortex was the source of tissue profiled in these studies, the relationship of miR-10b-5p expression to striatal involvement in the disease was independent of cortical involvement. Correlation of miRNAs to the clinical features clustered by direction of effect and the gene targets of the observed miRNAs showed association to processes relating to nervous system development and transcriptional regulation., Conclusions: These results demonstrate that miRNA expression in cortical BA9 provides insight into striatal involvement and support a role for these miRNAs, particularly miR-10b-5p, in HD pathogenicity. The miRNAs identified in our studies of postmortem brain tissue may be detectable in peripheral fluids and thus warrant consideration as accessible biomarkers for disease stage, rate of progression, and other important clinical characteristics of HD.

Aim: To evaluate the bonding temperature effect on dentin-restoration microleakage. The null hypothesis of the study is that the score of microleakage is identical among different adhesive bondings at different temperatures., Materials and Methods: Ninety caries free maxillary premolars were selected. Class V cavities were prepared on the cemento enamel junction (CEJ) of the buccal sides with enamel margins on occlusal sides and cementum margins on gingival sides. The specimens were divided into 3 groups: G1, single bond adhesive + Z250 composite; G2, P90 adhesive + Filtek Silorane composite; and G3, Clearfil SE bond + Clearfil APX. All groups were divided into three subgroups based on the adhesive temperature: A-4°C; B-25°C; and C-40°C. After coating the specimens with nail polish 1 mm beyond the margin of the restorations, they were stored in 0.5% basic Fuchsin dye solution for 24 hours. The teeth then were buccolingually sectioned and observed under a stereomicroscope., Results: There was no significant difference between microleakage of occlusal and gingival margins in each group. Clearfil SE bond and Adper single bond displayed lower microleakage than P90 adhesive at 4°C and 25°C. The most and least microleakage score for Adper single bond was at 40°C and 25°C respectively. Clearfil SE bond showed less microleakage at 25°C than 4°C and 40°C., Conclusion: Clearfil SE bond and Adper single bond displayed less microleakage at 25°C while there was no significant difference among for P90 adhesive microleakage at three temperatures.

To investigate epigenetic contributions to Huntington's disease (HD) pathogenesis, we carried out genome-wide mapping of the transcriptional mark, trimethyl-histone H3-lysine 4 (H3K4me3) in neuronal nuclei extracted from prefrontal cortex of HD cases and controls using chromatin immunoprecipitation followed by deep-sequencing. Neuron-specific mapping of the genome-wide distribution of H3K4me3 revealed 136 differentially enriched loci associated with genes implicated in neuronal development and neurodegeneration, including GPR3, TMEM106B, PDIA6 and the Notch signaling genes hairy and enhancer of split 4 (HES4) and JAGGED2, supporting the view that the neuronal epigenome is affected in HD. Importantly, loss of H3K4me3 at CpG-rich sequences on the HES4 promoter was associated with excessive DNA methylation, reduced binding of nuclear proteins to the methylated region and altered expression of HES4 and HES4 targeted genes MASH1 and P21 involved in striatal development. Moreover, hypermethylation of HES4 promoter sequences was strikingly correlated with measures of striatal degeneration and age-of-onset in a cohort of 25 HD brains (r = 0.56, P = 0.006). Lastly, shRNA knockdown of HES4 in human neuroblastoma cells altered MASH1 and P21 mRNA expression and markedly increased mutated HTT-induced aggregates and cell death. These findings, taken together, suggest that epigenetic dysregulation of HES4 could play a critical role in modifying HD disease pathogenesis and severity., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Purpose: To compare ocular biometric parameters using low-coherence interferometry among siblings affected with different degrees of primary angle closure (PAC)., Methods: In this cross-sectional comparative study, a total of 170 eyes of 86 siblings from 47 families underwent low-coherence interferometry (LenStar 900; Haag-Streit, Koeniz, Switzerland) to determine central corneal thickness, anterior chamber depth (ACD), aqueous depth (AD), lens thickness (LT), vitreous depth, and axial length (AL). Regression coefficients were applied to show the trend of the measured variables in different stages of angle closure. To evaluate the discriminative power of the parameters, receiver operating characteristic curves were used. Best cutoff points were selected based on the Youden index. Sensitivity, specificity, positive and negative predicative values, positive and negative likelihood ratios, and diagnostic accuracy were determined for each variable., Results: All biometric parameters changed significantly from normal eyes to PAC suspects, PAC, and PAC glaucoma; there was a significant stepwise decrease in central corneal thickness, ACD, AD, vitreous depth, and AL, and an increase in LT and LT/AL. Anterior chamber depth and AD had the best diagnostic power for detecting angle closure; best levels of sensitivity and specificity were obtained with cutoff values of 3.11 mm for ACD and 2.57 mm for AD., Conclusions: Biometric parameters measured by low-coherence interferometry demonstrated a significant and stepwise change among eyes affected with various degrees of angle closure. Although the current classification scheme for angle closure is based on anatomical features, it has excellent correlation with biometric parameters.

Prenatal protein malnutrition (PPM) in rats causes enduring changes in brain and behavior including increased cognitive rigidity and decreased inhibitory control. A preliminary gene microarray screen of PPM rat prefrontal cortex (PFC) identified alterations in KCNJ3 (GIRK1/Kir3.1), a gene important for regulating neuronal excitability. Follow-up with polymerase chain reaction and Western blot showed decreased KCNJ3 expression in the PFC, but not hippocampus or brainstem. To verify localization of the effect to the PFC, baseline regional brain activity was assessed with (14)C-2-deoxyglucose. Results showed decreased activation in the PFC but not hippocampus. Together these findings point to the unique vulnerability of the PFC to the nutritional insult during early brain development, with enduring effects in adulthood on KCNJ3 expression and baseline metabolic activity., (Copyright © 2014 IBRO. Published by Elsevier Ltd. All rights reserved.)

Purpose: To determine the frequency of angle closure in siblings of patients with primary angle-closure glaucoma (PACG)., Methods: In this cross-sectional observational study, siblings of consecutive patients with PACG underwent a comprehensive ophthalmologic examination, including slit-lamp biomicroscopy, measurement of intraocular pressure, stereoscopic optic nerve head examination, and dynamic gonioscopy. Ultrasonic pachymetry was obtained in all subjects; peripapillary retinal nerve fiber layer thickness measurement by optical coherence tomography and standard achromatic perimetry were performed in subjects with angle closure, categorized as primary angle-closure suspect (PACS), primary angle closure (PAC), and PACG, or any suspicion of glaucoma in the presence of open angles., Results: Overall, 95 siblings from 47 families with at least 1 subject affected with PACG participated in the evaluations; 55 (57.9%) individuals were categorized within the spectrum of angle closure, including 34 (35.8%) subjects with PACS, 7 (7.4%)individuals with PAC, and 14 (14.7%) cases of PACG. Nine (9.5%) individuals with open angles demonstrated other abnormal features, these included 3 (3.2%) subjects with suspicious discs, 5 (5.3%) cases of primary open-angle glaucoma, and 1 patient (1.1%) with normal-tension glaucoma. The remaining 31 individuals (32.6%) had no evidence of glaucoma., Conclusions: Our findings indicate familial segregation of angle closure. Siblings of PACG patients are at high risk for the condition, such that two thirds of them demonstrate clinical findings related to glaucoma. These observations suggest a hereditary basis for angle closure; therefore, siblings of patients affected with angle closure should undergo targeted screening for glaucoma.