The emulsifying and dispersing mechanisms of oil-in-water emulsions stabilized by 2,2,6,6-tetramethylpiperidin-1-oxyl (TEMPO)-oxidized cellulose nanofibers (CNFs) have been investigated. The ...emulsifying mechanism was studied by changing the oil/water interfacial tension from 8.5 to 53.3 mN/m using various types of oils. The results showed that the higher the oil/water interfacial tension, the greater is the amount of CNFs adsorbed at the oil/water interface, making the CNF-adsorbed oil-in-water emulsions thermodynamically more stable. Moreover, the amount of CNFs adsorbed on the surfaces of the oil droplets increased with increasing interfacial area. The dispersion stability of the oil droplets was dominated by the CNF concentration in the water phase. Above the critical concentration (0.15% w/w), the CNFs formed network structures in the water phase, and the emulsion was effectively stabilized against creaming. Emulsion formation and the CNF network structures in the emulsion were visualized by cryo-scanning electron microscopy.
After the Fukushima Daiichi Nuclear Power Plant accident, radiocesium, one of the main radioactive materials, has been accumulated inside konara oak trunks. Radiocesium has been thought to move ...radially through the trunk, but it has not been scientifically vindicated because the mechanism of the radial movement of minerals has not yet been experimentally determined. In this study, mineral radial movement was investigated in konara oak trunks of standing trees. A stable isotope cesium (Cs) solution was injected as a tracer into the outer sapwood of standing konara oak tree trunks. A trunk part was subsequently freeze-fixed with liquid nitrogen and subjected to Cs distribution analysis using cryo-scanning electron microscopy/energy-dispersive X-ray spectroscopy. By comparing normal samples that included living cells and freeze–thaw treated samples that contained no living cells, it was concluded that the injected Cs has been moving through the sapwood by the combination of rapid symplasmic movement by virtue of the living xylem parenchyma cells and slow apoplastic diffusion. Conversely, the Cs solution did not diffuse from the sapwood to the heartwood, implying that Cs is exuded from the living parenchyma cells to the apoplast, i.e., cell walls or adjoined dead parenchyma cells in the layer at the sapwood–heartwood boundary, and then diffused into the heartwood. By integrating the results of this study and our previous results obtained on Japanese cedar, we conclude that the mechanism of the radial movement of minerals through the sapwood seems to be a universal characteristic of tree species. In contrast, since mineral concentrations varied among tree species, the movement mechanism across the sapwood–heartwood boundary can differ among tree species.
Covalent modification of histones plays an important role in regulating chromatin dynamics and transcription. Histone methylation was thought to be an irreversible modification until recently. Using ...a biochemical assay coupled with chromatography, we have purified a JmjC domain-containing protein, JHDM2A, which specifically demethylates mono- and dimethyl-H3K9. Similar to JHDM1, JHDM2A-mediated histone demethylation requires cofactors Fe(II) and α-ketoglutarate. Mutational studies indicate that a JmjC domain and a zinc finger present in JHDM2A are required for its enzymatic activity. Overexpression of JHDM2A greatly reduced the H3K9 methylation level in vivo. Knockdown of JHDM2A results in an increase in the dimethyl-K9 levels at the promoter region of a subset of genes concomitant with decrease in their expression. Finally, JHDM2A exhibits hormone-dependent recruitment to androgen-receptor target genes, resulting in H3K9 demethylation and transcriptional activation. Thus, our work identifies a histone demethylase and links its function to hormone-dependent transcriptional activation.
Post-translational modification of chromatin has profound effects on many biological processes including transcriptional regulation, heterochromatin organization, and X-chromosome inactivation. ...Recent studies indicate that methylation on specific histone lysine (K) residues participates in many of these processes. Lysine methylation occurs in three distinct states, having either one (me1), two (me2) or three (me3) methyl groups attached to the amine group of the lysine side chain. These differences in modification state have an important role in defining how methylated chromatin is recognized and interpreted. Until recently, histone lysine methylation was considered a stable modification, but the identification of histone demethylase enzymes has demonstrated the reversibility of this epigenetic mark. So far, all characterized histone demethylases show enzymatic activity towards lysine residues modified in the me1 or me2 state, leaving open the possibility that me3 constitutes an irreversible modification. Here we demonstrate that JHDM3A (jumonji C (JmjC)-domain-containing histone demethylase 3A; also known as JMJD2A) is capable of removing the me3 group from modified H3 lysine 9 (H3K9) and H3 lysine 36 (H3K36). Overexpression of JHDM3A abrogates recruitment of HP1 (heterochromatin protein 1) to heterochromatin, indicating a role for JHDM3A in antagonizing methylated H3K9 nucleated events. siRNA-mediated knockdown of JHDM3A leads to increased levels of H3K9 methylation and upregulation of a JHDM3A target gene, ASCL2, indicating that JHDM3A may function in euchromatin to remove histone methylation marks that are associated with active transcription.
Key message
Ray parenchyma cells represent the main pathway for mineral radial movement from sapwood towards intermediate wood, based on direct cryo-scanning electron microscopy/energy dispersive ...X-ray spectroscopy analysis.
Ray parenchyma cells are considered to be associated with the radial movement of some elements from sapwood to heartwood in tree trunks. However, this function has not yet been experimentally confirmed. In this study, we analysed the radial movement of caesium (Cs), as a tracer of minerals, through the xylem at the cellular level in standing trees. A solution containing a stable Cs isotope diluted in aqueous acid fuchsin was injected into the outer sapwood of Japanese cedar trees for 3 h, 1, or 4 days. After the injection, the trunk (including the injection points) was freeze-fixed with liquid nitrogen. Subsequently, the trees were felled. The red colour from acid fuchsin was detected in the outer sapwood and did not move passively in the radial direction with free water. For detailed analysis of the Cs distribution in the radial direction (from the red-coloured sapwood towards the pith), we performed cryo-scanning electron microscopy/energy-dispersive X-ray spectroscopy point analysis of the frozen-hydrated samples to analyse the Cs distribution by detecting the characteristic Cs-Lα X-ray peak. In the outer sapwood, Cs was detected in all cell structures at all injection periods. In the inner xylem tissue, Cs was detected in the cell wall and lumen of ray parenchyma cells at 4-day injection, but not at 3-h and 1-day injection. These results suggest that ray parenchyma cells represent the main pathway whereby Cs can move radially from sapwood towards intermediate wood in standing Japanese cedar trees.
Lysine-specific demethylase 1 (LSD1) was recently identified as the first histone demethylase that specifically demethylates monomethylated and dimethylated histone H3 at K4. It is a component of the ...CoREST and other corepressor complexes and plays an important role in silencing neuronal-specific genes in nonneuronal cells, but the molecular mechanisms of its action remain unclear. The 2.8-Å-resolution crystal structure of the human LSD1 reveals that LSD1 defines a new subfamily of FAD-dependent oxidases. The active center of LSD1 is characterized by a remarkable$1,245-\circ{A}^{3}$substrate-binding cavity with a highly negative electrostatic potential. Although the protein core of LSD1 resembles other flavoenzymes, its enzymatic activity and functions require two additional structural modules: an N-terminal SWIRM domain important for protein stability and a large insertion in the catalytic domain indispensable both for the demethylase activity and the interaction with CoREST. These results provide a framework for further probing the catalytic mechanism and the functional roles of LSD1.
The radial movement of minerals in tree trunks is a widely accepted function of ray parenchyma cells, but there is little experimental evidence for this. We previously obtained experimental data ...showing that the parenchyma cells were the site of the radial mineral movement in Japanese cedar (Cryptomeria japonica D. Don) trunks in winter. Therefore, the aim of this study was to answer two remaining questions: do parenchyma cells move minerals via active transport or passive diffusion and how do seasonality and the injection duration affect the radial movement of minerals. To analyze this, we compared mineral movement in living standing Japanese cedar trees with heartwood in which the trunk had been left untreated or freeze–thawed with liquid nitrogen to kill the living cells. A solution of a stable isotope of cesium (Cs), as a tracer of mineral movement, was continuously injected into the outer sapwood of these normal and freeze–thaw-treated trees for an objective period, following which the trunk was freeze-fixed with liquid nitrogen. The Cs distribution in frozen samples was then analyzed by cryo-scanning electron microscopy/energy-dispersive X-ray spectroscopy. After 1 and 5 days of injection, the Cs detection area was almost the same among parenchyma cells and tracheid cell walls in the freeze–thaw-treated samples (without living cells) but was further toward the inner xylem in the parenchyma cells than the tracheids in the normal samples (with living cells), indicating that living parenchyma cells move Cs. Furthermore, after 5 days of injection, Cs in the tracheid cell walls was detected further toward the inner xylem in the normal samples than in the freeze–thaw-treated samples, indicating that Cs is exuded from the parenchyma cells into the tracheid cell walls. Together, these results suggest that the radial movement of minerals in standing Japanese cedar trees occurs through a combination of active transport by parenchyma cells and diffusion in the cell walls.
Iron- and 2-oxoglutarate-dependent dioxygenases are a diverse family of non-heme iron enzymes that catalyze various important oxidations in cells. A key structural motif of these dioxygenases is a ...facial triad of 2-histidines-1-carboxylate that coordinates the Fe(II) at the catalytic site. Using histone demethylase JMJD1A and DNA repair enzyme ABH2 as examples, we show that this family of dioxygenases is highly sensitive to inhibition by carcinogenic nickel ions. We find that, with iron, the 50% inhibitory concentrations of nickel (IC50 Ni(II)) are 25 μm for JMJD1A and 7.5 μm for ABH2. Without iron, JMJD1A is 10 times more sensitive to nickel inhibition with an IC50 Ni(II) of 2.5 μm, and approximately one molecule of Ni(II) inhibits one molecule of JMJD1A, suggesting that nickel causes inhibition by replacing the iron. Furthermore, nickel-bound JMJD1A is not reactivated by excessive iron even up to a 2 mm concentration. Using x-ray absorption spectroscopy, we demonstrate that nickel binds to the same site in ABH2 as iron, and replacement of the iron by nickel does not prevent the binding of the cofactor 2-oxoglutarate. Finally, we show that nickel ions target and inhibit JMJD1A in intact cells, and disruption of the iron-binding site decreases binding of nickel ions to ABH2 in intact cells. Together, our results reveal that the members of this dioxygenase family are specific targets for nickel ions in cells. Inhibition of these dioxygenases by nickel is likely to have widespread impacts on cells (e.g. impaired epigenetic programs and DNA repair) and may eventually lead to cancer development.
This study sought to identify subgroups of attention-deficit hyperactivity disorder (ADHD) defined by specific patterns of emotional and behavioral symptoms according to the parent-rated Child ...Behavior Checklist (CBCL). Our clinical sample comprised 314 children (aged 4 to 15 years) diagnosed with ADHD according to the DSM-5. In addition, comorbid psychiatric disorders, general functioning, and medication status were assessed. Cluster analysis was performed on the CBCL syndrome subscales and yielded a solution with four distinct subgroups. The “High internalizing/externalizing” group displayed an overlap between internalizing and externalizing problems in the CBCL profile. In addition, the “High internalizing/externalizing” group revealed a high rate of comorbid autism spectrum disorder and elevated autistic traits. The “Inattention and internalizing” group revealed a high rate of the predominantly inattentive presentation according to ADHD specifier from the DSM-5. The “Aggression and externalizing” group revealed a high rate of comorbid oppositional defiant disorder and conduct disorder. The “Less psychopathology” group scored low on all syndrome scales. Children with ADHD were subdivided into four distinct subgroups characterized by psychopathological patterns, with and without internalizing and externalizing problems. The overlap between internalizing and externalizing problems may be mediated with emotional dysregulation and associated neurobiological bases.
Changes in histone methylation status regulate chromatin structure and DNA-dependent processes such as transcription. Recent studies indicate that, analogous to other histone modifications, histone ...methylation is reversible. Retinoblastoma binding protein 2 (RBP2), a nuclear protein implicated in the regulation of transcription and differentiation by the retinoblastoma tumor suppressor protein, contains a JmjC domain recently defined as a histone demethylase signature motif. Here we report that RBP2 is a demethylase that specifically catalyzes demethylation on H3K4, whose methylation is normally associated with transcriptionally active genes.
RBP2−/− mouse cells displayed enhanced transcription of certain cytokine genes, which, in the case of
SDF1, was associated with increased H3K4 trimethylation. Furthermore, RBP2 specifically demethylated H3K4 in biochemical and cell-based assays. These studies provide mechanistic insights into transcriptional regulation by RBP2 and provide the first example of a mammalian enzyme capable of erasing trimethylated H3K4.
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