Inhibition of DNA methyltransferase leads to increased genomic 5-hydroxymethylcytosine levels in hematopoietic cells.

5-Hydroxymethylcytosine (5hmC) is produced from 5-methylcytosine (5mC) by Ten-eleven translocation (TET) dioxygenases. The epigenetic modification 5hmC has crucial roles in both cellular development and differentiation. The 5hmC level is particularly high in the brain. While 5mC is generally associated with gene silencing/reduced expression, 5hmC is a more permissive epigenetic mark. To understand its physiological function, an easy and accurate quantification method is required. Here, we have developed a novel LC-MS/MS-based approach to quantify both genomic 5mC and 5hmC contents. The method is based on the liberation of nucleobases by formic acid. Applying this method, we characterized the levels of DNA methylation and hydroxymethylation in mouse brain and liver, primary hepatocytes, and various cell lines. Using this approach, we confirm that the treatment of different cell lines with the DNA methyltransferase inhibitor 5-aza-2\u27-deoxycytidine leads to a decrease in 5mC content. This decrease was accompanied by an increase in 5hmC levels in cell lines of hematopoietic origin. Finally, we showed that ascorbate elevates the levels of 5hmC and augments the effect of 5-aza-2\u27-deoxycytidine without significantly influencing 5mC levels

Reliability of Reward ERPs in Middle-Late Adolescents Using a Custom and a Standardized Preprocessing Pipeline

Despite advantage of neuroimaging measures in translational research frameworks, less is known about the psychometric properties thereof, especially in middle‐late adolescents. Earlier, we examined evidence of convergent and incremental validity of reward anticipation and response event‐related potentials (ERPs) and here we examined, in the same sample of 43 adolescents (M (age) = 15.67 years; SD = 1.01; range: 14–18; 32.6% boys), data quality (signal‐to‐noise ratio [SNR]), stability (mean amplitude across trials), and internal consistency (Cronbach’s α and split‐half reliability) of the same ERPs. Further, because observed time course and peak amplitude of ERP grand averages and thus findings on SNR, stability, and internal consistency may depend on preprocessing method, we employed a custom and a standardized preprocessing pipeline and compared findings across those. Using our custom pipeline, reward anticipation components were stable by the 40th trial, achieved acceptable internal consistency by the 19th, and all (but the stimulus‐preceding negativity [SPN]) achieved acceptable SNR by the 41st trial. Initial response to reward components were stable by the 20th trial and achieved acceptable internal consistency by the 11th and acceptable SNR by the 45th trial. Difference scores had worse psychometric properties than parent measures. Time course and peak amplitudes of ERPs and thus results on SNR, stability, and internal consistency were comparable across preprocessing pipelines. In case of reward anticipation ERPs examined here, 41 trials (+4 artifacted and removed) and, in case of reward response ERPs, 45 trials (+5 artifacted) yielded stable and internally consistent estimates with acceptable SNR. Results are robust across preprocessing methods

Galanin-immunoreactivity identifies a distinct population of inhibitory interneurons in laminae I-III of the rat spinal cord

Background: Inhibitory interneurons constitute 30-40% of neurons in laminae I-III and have an important anti-nociceptive role. However, because of the difficulty in classifying them we know little about their organisation. Previous studies have identified 3 non-overlapping groups of inhibitory interneuron, which contain neuropeptide Y (NPY), neuronal nitric oxide synthase (nNOS) or parvalbumin, and have shown that these differ in postsynaptic targets. Some inhibitory interneurons contain galanin and the first aim of this study was to determine whether these form a different population from those containing NPY, nNOS or parvalbumin. We also estimated the proportion of neurons and GABAergic axons that contain galanin in laminae I-III. Results: Galanin cells were concentrated in laminae I-IIo, with few in laminae IIi-III. Galanin showed minimal co-localisation with NPY, nNOS or parvalbumin in laminae I-II, but most galanin-containing cells in lamina III were nNOS-positive. Galanin cells constituted similar to 7%, 3% and 2% of all neurons in laminae I, II and III, and we estimate that this corresponds to 26%, 10% and 5% of the GABAergic neurons in these laminae. However, galanin was only found in similar to 6% of GABAergic boutons in laminae I-IIo, and similar to 1% of those in laminae IIi-III. Conclusions: These results show that galanin, NPY, nNOS and parvalbumin can be used to define four distinct neurochemical populations of inhibitory interneurons. Together with results of a recent study, they suggest that the galanin and NPY populations account for around half of the inhibitory interneurons in lamina I and a quarter of those in lamina I

A koherencia mint a lelki és testi egészség alapvető meghatározója a mai magyar társadalomban = Sense of coherence as an important determinant of mental and physical health

Az ún. salutogenezis modell olyan keretet kínál, amelyben a koherencia élmény bevezetésével lehetőség nyílik az „egész”-ség dinamikus értelmezésére. Vizsgálatunkban a Richard Rahe-féle, az „élet értelme” koherencia kérdőív összefüggéseit vizsgáltuk az egészségi állapottal. A Hungarostudy 2002 felmérés a 18 évesnél idősebb magyar népességet életkor, nem és terület szerint képviseli. 12 640 személlyel vettünk fel otthoni interjút. Az így vizsgált koherencia mutatót az egészségi állapot igen fontos előrejelzőjének találtuk. Ha az adatokat életkor, nem és iskolázottság szerint korrigáltuk, az egészségi állapot önbecslése mintegy 10-szer, a munkaképesség 8-szor jobb, a depresszió valószínűsége 7-szer alacsonyabb volt. Az „élet értelme” mutató igen szoros kapcsolatban áll az önhatékonysággal, a problémaorientált megbirkózással, a társas támogatással, ezzel szemben kevésbé függ az iskolázottságtól, az életkortól és a nemtől

Genome-wide association analysis reveals KCTD12 and miR-383-binding genes in the background of rumination

Ruminative response style is a passive and repetitive way of responding to stress, associated with several disorders. Although twin and candidate gene studies have proven the genetic underpinnings of rumination, no genome-wide association study (GWAS) has been conducted yet. We performed a GWAS on ruminative response style and its two subtypes, brooding and reflection, among 1758 European adults recruited in the general population of Budapest, Hungary, and Manchester, United Kingdom. We evaluated single-nucleotide polymorphism (SNP)-based, gene-based and gene set-based tests, together with inferences on genes regulated by our most significant SNPs. While no genome-wide significant hit emerged at the SNP level, the association of rumination survived correction for multiple testing with KCTD12 at the gene level, and with the set of genes binding miR-383 at the gene set level. SNP-level results were concordant between the Budapest and Manchester subsamples for all three rumination phenotypes. SNPlevel results and their links to brain expression levels based on external databases supported the role of KCTD12, SRGAP3, and SETD5 in rumination, CDH12 in brooding, and DPYSL5, MAPRE3, KCNK3, ATXN7L3B, and TPH2 in reflection, among others. The relatively low sample size is a limitation of our study. Results of the first GWAS on rumination identified genes previously implicated in psychiatric disorders underscoring the transdiagnostic nature of rumination, and pointed to the possible role of the dorsolateral prefrontal cortex, hippocampus, and cerebellum in this cognitive process

Recommendations, guidelines, and best practice for the use of human induced pluripotent stem cells for neuropharmacological studies of neuropsychiatric disorders

The number of individuals suffering from neuropsychiatric disorders (NPDs) has increased worldwide, with 3 million disability-adjusted life-years calculated in 2019. Though research using various approaches including genetics, imaging, clinical and animal models has advanced our knowledge regarding NPDs, we still lack basic knowledge regarding the underlying pathophysiological mechanisms. Moreover, there is an urgent need for highly effective therapeutics for NPDs. Human induced pluripotent stem cells (hiPSCs) generated from somatic cells enabled scientists to create brain cells in a patient-specific manner. However, there are challenges to the use of hiPSCs that need to be addressed. In the current paper, consideration of best practices for neuropharmacological and neuropsychiatric research using hiPSCs will be discussed. Specifically, we provide recommendations for best practice in patient recruitment, including collecting demographic, clinical, medical (before and after treatment and response), diagnostic (including scales) and genetic data from the donors. We highlight considerations regarding donor genetics and sex, in addition to discussing biological and technical replicates. Furthermore, we present our views on selecting control groups/lines, experimental designs, and considerations for conducting neuropharmacological studies using hiPSC-based models in the context of NPDs. In doing so, we explore key issues in the field concerning reproducibility, statistical analysis, and how to translate in vitro studies into clinically relevant observations. The aim of this article is to provide a key resource for hiPSC researchers to perform robust and reproducible neuropharmacological studies, with the ultimate aim of improving identification and clinical translation of novel therapeutic drugs for NPDs

Live fast, die young? A review on the developmental trajectories of ADHD across the lifespan

Attention-deficit/hyperactivity disorder (ADHD) is highly heritable and the most common neurodevelopmental disorder in childhood. In recent decades, it has been appreciated that in a substantial number of cases the disorder does not remit in puberty, but persists into adulthood. Both in childhood and adulthood, ADHD is characterised by substantial comorbidity including substance use, depression, anxiety, and accidents. However, course and symptoms of the disorder and the comorbidities may fluctuate and change over time, and even age of onset in childhood has recently been questioned. Available evidence to date is poor and largely inconsistent with regard to the predictors of persistence versus remittance. Likewise, the development of comorbid disorders cannot be foreseen early on, hampering preventive measures. These facts call for a lifespan perspective on ADHD from childhood to old age. In this selective review, we summarise current knowledge of the long-term course of ADHD, with an emphasis on clinical symptom and cognitive trajectories, treatment effects over the lifespan, and the development of comorbidities. Also, we summarise current knowledge and important unresolved issues on biological factors underlying different ADHD trajectories. We conclude that a severe lack of knowledge on lifespan aspects in ADHD still exists for nearly every aspect reviewed. We encourage large-scale research efforts to overcome those knowledge gaps through appropriately granular longitudinal studies