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Background Kabuki syndrome (KS) is a rare developmental disorder characterised by multiple congenital anomalies and intellectual disability. UTX (ubiquitously transcribed tetratricopeptide repeat, X chromosome), which encodes a histone demethylase, is one of the two major pathogenic risk genes for KS. Although intellectual disability is a key phenotype of KS, the role of UTX in cognitive function remains unclear. Currently, no targeted therapies are available for KS.Aims This study aimed to investigate how UTX regulates cognition, to explore the mechanisms underlying UTX dysfunction and to identify potential molecular targets for treatment.Methods We generated UTX conditional knockout mice and found that UTX deletion downregulated calmodulin transcription by disrupting H3K27me3 (trimethylated histone H3 at lysine 27) demethylation.Results UTX-knockout mice showed decreased phosphorylation of calcium / calmodulin-dependent protein kinase II, impaired long-term potentiation and deficit in remote contextual fear memory. These effects were reversed by an Food and Drug Administration-approved drug desipramine.Conclusions Our results reveal an epigenetic mechanism underlying the important role of UTX in synaptic plasticity and cognitive function, and suggest that desipramine could be a potential treatment for KS.

The mammalian epididymis not only plays a fundamental role in the maturation of spermatozoa, but also provides protection against various stressors. The foremost among these is the threat posed by oxidative stress, which arises from an imbalance in reactive oxygen species and can elicit damage to cellular lipids, proteins, and nucleic acids. In mice, the risk of oxidative damage to spermatozoa is mitigated through the expression and secretion of glutathione peroxidase 5 (GPX5) as a major luminal scavenger in the proximal caput epididymidal segment. Accordingly, the loss of GPX5-mediated protection leads to impaired DNA integrity in the spermatozoa of aged Gpx5-/- mice. To explore the underlying mechanism, we have conducted transcriptomic analysis of caput epididymidal epithelial cells from aged (13 months old) Gpx5-/- mice. This analysis revealed the dysregulation of several thousand epididymal mRNA transcripts, including the downregulation of a subgroup of piRNA pathway genes, in aged Gpx5-/- mice. In agreement with these findings, we also observed the loss of piRNAs, which potentially bind to the P-element-induced wimpy testis (PIWI)-like proteins PIWIL1 and PIWIL2. The absence of these piRNAs was correlated with the elevated mRNA levels of their putative gene targets in the caput epididymidis of Gpx5-/- mice. Importantly, the oxidative stress response genes tend to have more targeting piRNAs, and many of them were among the top increased genes upon the loss of GPX5. Taken together, our findings suggest the existence of a previously uncharacterized somatic piRNA pathway in the mammalian epididymis and its possible involvement in the aging and oxidative stress-mediated responses.

Body axial patterning develops via a rostral-to-caudal sequence and relies on the temporal colinear activation of Hox genes. However, the underlying mechanism of Hox gene temporal colinear activation remains largely elusive. Here, with small-molecule inhibitors and conditional gene knockout mice, we identified Jmjd3, a subunit of TrxG, as an essential regulator of temporal colinear activation of Hox genes with its H3K27me3 demethylase activity. We demonstrated that Jmjd3 not only initiates but also maintains the temporal collinear expression of Hox genes. However, we detected no antagonistic roles between Jmjd3 and Ezh2, a core subunit of PcG repressive complex 2, during the processes of axial skeletal patterning. Our findings provide new insights into the regulation of Hox gene temporal collinear activation for body axial patterning in mice.

Summary: Histone methyl groups can be removed by demethylases. Although LSD1 and JmjC domain-containing proteins have been identified as histone demethylases, enzymes for many histone methylation states or sites are still unknown. Here, we perform a screening of a cDNA library containing 2,500 nuclear proteins and identify hHR23A as a histone H4K20 demethylase. Overexpression of hHR23A reduces the levels of H4K20me1/2/3 in cells. In vitro, hHR23A specifically demethylates H4K20me1/2/3 and generates formaldehyde. The enzymatic activity requires Fe(II) and α-ketoglutarate as cofactors and the UBA domains of hHR23A. hHR23B, a protein homologous to hHR23A, also demethylates H4K20me1/2/3 in vitro and in vivo. We further demonstrate that hHR23A/B activate the transcription of coding genes by demethylating H4K20me1 and the transcription of repetitive elements by demethylating H4K20me3. Nuclear magnetic resonance (NMR) analyses demonstrate that an HxxxE motif in the UBA1 domain is crucial for iron binding and demethylase activity. Thus, we identify two hHR23 proteins as histone demethylases. : Cao et al. screen 2,500 nuclear proteins and find that hHR23A/B are histone H4K20 demethylases. They also show that hHR23A/B activate transcription via demethylation. Keywords: histone demethylase, H4K20 demethylase, H4K20 methylation, UBA domain-containing proteins

UTX is a tumor suppressor gene located on the X-chromosome so it could potentially contribute to the cancer gender bias. Here the authors, using a mouse model of B cell lymphoma, show that UTX is a dosage sensitive tumor suppressor and may be responsible for some of the increased incidence and possibly aggressiveness of male cancers that harbour UTX mutations.

Mutations in PHF8 gene are genetically associated with X-linked mental retardation. Here, Chen et al. show that Phf8 KO mouse have cognitive and synaptic plasticity impairment, and pharmacological inhibition of mTOR signaling can partially alleviate such defects.

Epigenomic abnormalities caused by genetic mutation in epigenetic regulators can result in neurodevelopmental disorders, deficiency in neural plasticity and mental retardation. As a histone demethylase, plant homeodomain finger protein 8 (Phf8) is a candidate gene for syndromal and non-specific forms of X-chromosome-linked intellectual disability (XLID). Here we report that Phf8 knockout mice displayed impaired learning and memory, and impaired hippocampal long-term potentiation (LTP) without gross morphological defects. We also show that mTOR signaling pathway is hyperactive in hippocampus in Phf8 knockout mouse. Mechanistically, we show that demethylation of H4K20me1 by Phf8 results in transcriptional suppression of RSK1 and homeostasis of mTOR signaling. Pharmacological suppression of mTOR signaling with rapamycin in Phf8 knockout mice recovers the weakened LTP and cognitive deficits. Together, our results indicate that loss of Phf8 in animals causes deficient learning and memory by epigenetic disruption of mTOR signaling, and provides a potential therapeutic drug target to treat XLID. [ABSTRACT FROM AUTHOR]

Interleukin (IL) 17-producing T helper (Th17) cells play critical roles in the clearance of extracellular bacteria and fungi as well as the pathogenesis of various autoimmune diseases, such as multiple sclerosis, psoriasis, and ulcerative colitis. Although a global transcrip-tional regulatory network of Th17 cell differentiation has been mapped recently, the participation of epigenetic modifications in the differentiation process has yet to be elucidated. We demonstrated here that histone H3 lysine-27 (H3K27) demethylation, predominantly mediated by the H3K27 demethylase Jmjd3, crucially regulated Th17 cell differentiation. Activation of naive CD4+ T cells immediately induced high expression of Jmjd3. Genetic depletion of Jmjd3 in CD4+ T cells specifically impaired Th17 cell differentiation both in vitro and in vivo. Ectopic expression of Jmjd3 largely rescued the impaired differentiation of Th17 cells in vitro in Jmjd3-deficient CD4+ T cells. Importantly, Jmjd3-deficient mice were resistant to the induction of experimental autoimmune encephalomyelitis (EAE). Furthermore, inhibition of the H3K27 demethylase activity with the specific inhibitor GSK-J4 dramatically suppressed Th17 cell differentiation in vitro. At the molecular level, Jmjd3 directly bound to and reduced the level of H3K27 trimethylation (me3) at the genomic sites of Rorc, which encodes the master Th17 transcription factor RorΥt, and Th17 cytokine genes such as Il17, Il17f, and Il22. Therefore, our studies established a critical role of Jmjd3-mediated H3K27 demethylation in Th17 cell differentiation and suggest that Jmjd3 can be a novel therapeutic target for suppressing autoimmune responses. [ABSTRACT FROM AUTHOR]

Aimed to provide a more close-to-reality and rapid calculation method for optimization design of low voltage circuit breaker with complex mechanism, an interruption model is constructed, which can describe the coupling of air blast process with arc model, complex mechanical movement, magnetic field, and electric circuit. The model is based on the non-stationary gas conservation equations, and considers the influence of the chamber structure on the gas dynamics. To realize the interaction between the arc movement and the gas flow simply, on the one hand, the moving electric arc is deemed an energy input source of the chamber gas. On the other hand, the movement equation of the arc considering action of the gas flow is built. With this model, the general principle of the chamber gas flow, arc voltage and current during the interruption process of an actual breaker are calculated and compared with the experiment results. As an application, the influence of the chamber vent size on the arc movement is studied with the model as well. [ABSTRACT FROM AUTHOR]

Introduction: Histone H3 lysine 27 trimethylation (H3K27me3) and H3 lysine 36 trimethylation (H3K36me3) are important epigenetic modifications correlated with transcription repression and activation, respectively. These two opposing modifications rarely co-exist in the same H3 polypeptide. However, a small but significant amount of H3 tails are modified with 5 methyl groups on K27 and K36 in mouse embryonic stem cells (mESCs) and it is unclear how the trimethylation is distributed on K27 or K36. Methods: A label-free, bottom-up mass spectrum method, named specific ions of isobaric modification chromatogram (SIMC), was established to quantify the relative abundance of K27me2-K36me3 and K27me3-K36me2 in the same histone H3 tail. Results: By using this method, we demonstrated that the H3K27me3-K36me2 comprises about 85 % of the penta-methylated H3 tails at K27 and K36 in mESCs. Upon mESC differentiation, the abundance of H3K27me3-K36me2 significantly decreased, while the level of H3K27me2-K36me3 remains unchanged. Conclusion: Our study not only revealed the cis-existence of H3K27me3-K36me2 in mESCs, but also suggested that this combinatorial histone modification may assume a specific regulatory function during differentiation. [ABSTRACT FROM AUTHOR]

Although it is well established that embryonic stem (ES) cells have the potential to differentiate into dopamine neurons, the molecular basis of this process, particularly the role of microRNAs (miRNAs), remains largely unknown. Here we report that miR-132 plays a key role in the differentiation of dopamine neurons by directly regulating the expression of Nurr1 (also known as nuclear receptor subfamily 4 group A member 2; Nr4a2). We constructed a mouse ES cell line CGR8, which stably expresses GFP under the tyrosine hydroxylase (TH) promoter, so the TH-positive neurons could be easily sorted using fluorescence-activated cell sorting (FACS). Then, we performed a miRNA array analysis on the purified TH-positive neurons and found that 45 of 585 miRNAs had more than a fivefold change in expression level during dopamine neuron differentiation. Among the 45 miRNAs, we were particularly interested in miR-132 because this miRNA has been reported to be highly expressed in neurons and to have a potential role in neurodegenerative diseases. We found that the direct downregulation of endogenous miR-132 induced by miR-132 antisense oligonucleotide (miR-132-ASO) promoted the differentiation of TH-positive neurons, whereas ectopic expression of miR-132 in ES cells reduced the number of differentiated THpositive neurons but did not change the total number of differentiated neurons. Furthermore, we identified that miR-132-ASO could substantially reverse the miR-132-mediated suppression of TH-positive neuron differentiation. Moreover, through a bioinformatics assay we identified the Nurr1 gene as a potential molecular target of miR-132. Using a luciferase-reporter assay and western blot analysis, we demonstrated that miR-132 could directly regulate the expression of Nurr1. Collectively, our data provide the first evidence that miR-132 is an important molecule regulating ES cell differentiation into dopamine neurons by directly targeting Nurr1 gene expression. [ABSTRACT FROM AUTHOR]

Dimethylation of histone H3 lysine 9 (H3K9me2) is an important epigenetic mark associated with transcription repression. Here, we identified PHF8, a JmjC-domain-containing protein, as a histone demethylase specific for this repressing mark. Recombinant full-length wild type protein could remove methylation from H3K9me2, but mutation of a conserved histidine to alanine H247A abolished the demethylase activity. Overexpressed exogenous PHF8 was colocalized with B23 staining. Endogenous PHF8 was also colocalized with B23 and fibrillarin, two well-established nucleolus proteins, suggesting that PHF8 is localized in the nucleolus and may regulate rRNA transcription. Indeed, PHF8 bound to the promoter region of the rDNA gene. Knockdown of PHF8 reduced the expression of rRNA, and overexpression of the gene resulted in upregulation of rRNA transcript. Concomitantly, H3K9me2 level was elevated in the promoter region of the rDNA gene in PHF8 knockdown cells and reduced significantly when the wild type but not the catalytically inactive H247A mutant PHF8 was overexpressed. Thus, our study identified a histone demethylase for H3K9me2 that regulates rRNA transcription. [ABSTRACT FROM AUTHOR]

Dimethylations of histone H3 lysine 9 and lysine 27 are important epigenetic marks associated with transcription repression. Here, we identified KIAA1718 (KDM7A) as a novel histone demethylase specific for these two repressing marks. Using mouse embryonic stem cells, we demonstrated that KIAA1718 expression increased at the early phase of neural differentiation. Knockdown of the gene blocked neural differentiation and the effect was rescued by the wild-type human gene, and not by a catalytically inactive mutant. In addition, overexpression of KIAA1718 accelerated neural differentiation. We provide the evidence that the pro-neural differentiation effect of KDM7A is mediated through direct transcriptional activation of FGF4, a signal molecule implicated in neural differentiation. Thus, our study identified a dual-specificity histone demethylase that regulates neural differentiation through FGF4. [ABSTRACT FROM AUTHOR]

This paper is devoted to study the influence of ferromagnetic material on arc behavior in low voltage electric apparatus. With the self-programmed interface, one 2-D magnetohydrodynamic arc model can be developed based on the commercial software FLUENT and ANSYS, to study the vapor of ferromagnetic material effects on air arc behavior. Then, the corresponding interruption experiments are performed to verify the simulation qualitatively. It demonstrates that the vapor of ferromagnetic material is disadvantageous to arc interruption characteristics. In addition, the influence of the size of the ferromagnetic splitter plate on arc motion is simulated. It shows that the bigger splitter plate may accelerate the arc motion. [ABSTRACT FROM AUTHOR]

Histone methylation is a dynamic process that participates in a diverse array of cellular processes and has been found to associate with cancer. Recently, several histone demethylases have been identified that catalyze the removal of methylation from histone H3 lysine residues. Through bioinformatic and biochemical analysis, we identified JARID1B as a H3K4 demethylase. Overexpression of JARID1B resulted in loss of tn-, di-, and monomethyl H3K4 but did not affect other histone lysine methylations. In vitro biochemical experiments demonstrated that JARID1B directly catalyzes the demethylation. The enzymatic activity requires the JmjC domain and uses Fe(II) and a-ketoglutarate as cofactors. Furthermore, we found that JARID1B is up-regulated in prostate cancer tissues, compared with benign prostate samples. We also demonstrated that JARID1B associates with androgen receptor and regulates its transcriptional activity. Thus, we identified JARID1B as a demethylase capable of removing three methyl groups from histone H3 lysine 4 and up-regulated in prostate cancer. [ABSTRACT FROM AUTHOR]

A two-dimensional arc magnetohydrodynamic model is presented to study the effects of metallic vapour and gassing material on air switching arc motion based on different mixture plasma properties (pure air, 99% air-1% Cu, 99% air-1% Ag, 99% air-1% Fe, 90% air-10% PA6). It demonstrates that metallic vapours may cool the arc central region and reduce the arc motion velocity, and iron vapours have the strongest action compared with copper and silver vapours; the gassing material may improve the arc motion velocity. In addition, the effect of the net emission coefficient model and the grey body radiation model on the calculation results for pure air arc motion is compared. To verify the simulation results partially, using the integrated imaging diagnostics system, experiments are carried out to measure the average velocity of arc motion with one model chamber. [ABSTRACT FROM AUTHOR]

To the optimization design of molded case circuit breakers (MCCBs), it is important and necessary to simulate the interruption process and analyze the characteristics. A set of differential equations can describe the interactive coupled phenomena of electric circuit, electromagnetic field and mechanism motion system in MCCB. First, with virtual prototyping technology, the simulation model for the operation mechanism of MCCB can be built, and experiments have been carried out to verify its validity. Then, with the existence of electrodynamic repulsion force, the interruption process of the MCCB has been investigated with the influence of arc voltage, closing phase angle, prospective current, mechanism starting motion time and blow open force. It demonstrates the proposed method is effective and is capable of evaluating the new design of MCCB products. [ABSTRACT FROM AUTHOR]

Arc-motion process is crucial to the interruption capability of the low-voltage circuit breaker. First, to a simplified chamber with full open venting, with a 2-D optical-fiber arc-motion measurement system, experiments were done to measure the arc-motion velocity in order to analyze the influence of different arc ignition position on the arc motion. It demonstrates that the larger the volume behind the arc is, the slower the arc moves. In addition, the influence of the gassing material on arc-motion velocity was also studied experimentally. Then, the commercial computational fluid dynamics software FLUENT was adapted and modified to develop a 2-D magneto-hydrodynamic model of the arc with the same conditions as the experimental model. The simulation result was consistent with the experiment conclusions. [ABSTRACT FROM AUTHOR]

To the optimization design of molded case circuit breakers (MCCBs), it is necessary and important to calculate the electro-dynamic repulsion force acting on the movable conductor. With three-dimensional (3-D) finite element nonlinear analysis, according to the equations among current-magnetic field-repulsion force and taking into account the ferromagnet, contact bridge model is introduced to simulate the current constriction between contacts, so Lorentz and Holm force acting on the movable conductor and contact, respectively, can be combined to calculate. Coupled with circuit equations, the opening time of movable contact also can be obtained using iteration with the restriction of contact force. Simulation and experiment for repulsion force and opening time of five different configuration models have been investigated. The results indicate that the proposed method is effective and capable of evaluating new design of contact systems in MCCBs. [ABSTRACT FROM AUTHOR]

This paper is devoted to the study of the air arc plasma characteristics and consists of two parts. An optical high-speed imaging system is integrated with pressure sensing, arc current, and voltage measurement to investigate high-speed and high-temperature unsteady arc plasma motion in a molded case circuit breaker (MCCB) model, especially with the influence of polyoxymethylene (POM), polycarbonate (PBT), nylon, and ceramic arc chamber wall materials. It demonstrates that the performance can be improved effectively by adopting POM and nylon gassing materials. Also, spectrum diagnostics is introduced to analyze the influence of polymethylmethacrylate (PMMA) gassing material on the temperature of arc in a simple model. It shows that material added adjacent to cathode is more effective to cool the arc, compared with the case of anode. [ABSTRACT FROM AUTHOR]