Revealing the Mechanism of Xist-mediated Silencing

Xist initiates XCI by spreading across the future inactive X-chromosome, excluding RNA polymerase II, recruiting the polycomb repressive complex and its associated repressive chromatin modifications, and repositioning active genes into a transcriptionally silenced nuclear compartment. While much is known about the events that occur during XCI, the mechanism by which Xist carries out these various roles remains unclear. Here we identify ten proteins that directly associate with Xist, and we further show that three of these proteins are required for Xist-mediated transcriptional silencing. One of these proteins, SHARP, which is known to interact with the SMRT co-repressor that activates HDAC3, is not only essential for silencing, but is also required for the exclusion of PolII from the inactive X. We show that both SMRT and HDAC3 are required for Xist-mediated silencing and RNA polymerase II exclusion. Another of these proteins, LBR, is required for repositioning actively transcribed genes into the Xist-silenced compartment. We further show that Xist, through its interaction with LBR, a protein that is anchored in the inner nuclear membrane, would effectively reposition Xist-coated DNA to the nuclear lamina, thereby changing the accessibility of other genes on the X-chromosome to enable Xist to spread to active genes across the entire chromosome to silence chromosome-wide transcription. Together, these results present an integrative picture of how Xist can scaffold multiple proteins to orchestrate the complex functions required for the establishment of the inactive X-chromosome.</p

A study of the total chromatic number of equibipartite graphs

AbstractThe total chromatic number χt(G) of a graph G is the least number of colors needed to color the vertices and edges of G so that no adjacent vertices or edges receive the same color, no incident edges receive the same color as either of the vertices it is incident with. In this paper, we obtain some results of the total chromatic number of the equibipartite graphs of order 2n with maximum degree n − 1. As a part of our results, we disprove the biconformability conjecture

A study of the total chromatic number of equibipartite graphs

AbstractThe total chromatic number χt(G) of a graph G is the least number of colors needed to color the vertices and edges of G so that no adjacent vertices or edges receive the same color, no incident edges receive the same color as either of the vertices it is incident with. In this paper, we obtain some results of the total chromatic number of the equibipartite graphs of order 2n with maximum degree n − 1. As a part of our results, we disprove the biconformability conjecture

High-Reliability Trigate Poly-Si Channel Flash Memory Cell With Si-Nanocrystal Embedded Charge-Trapping Layer

Abstract-This letter introduces a polycrystalline-silicon nanowire (NW) thin-film nonvolatile memory (NVM) with a self-assembled silicon-nanocrystal (Si-NC) embedded chargetrapping (CT) layer. This process is simple and compatible with conventional CMOS processes. Experimental results indicate that this NW NVM exhibits high reliability due to a deep-quantum-well structure and immunity of enhanced electric field underneath a disk-shaped Si-NC. After 10 000 P/E cycles, the memory window loss of the NVM with a Si-NC embedded CT layer is less than 12% until 10 4 s at 150 • C. Accordingly, a poly-Si thin-film transistor with a Si-NC embedded CT layer is highly promising for NVM applications. Index Terms-Nanocrystal (NC), nonvolatile memory (NVM), thin-film transistor (TFT)

Implications for Chk1 Regulation: The 1.7 Å Crystal Structure of Human Cell Cycle Checkpoint Kinase Chk1

AbstractThe checkpoint kinase Chk1 is an important mediator of cell cycle arrest following DNA damage. The 1.7 Å resolution crystal structures of the human Chk1 kinase domain and its binary complex with an ATP analog has revealed an identical open kinase conformation. The secondary structure and side chain interactions stabilize the activation loop of Chk1 and enable kinase activity without phosphorylation of the catalytic domain. Molecular modeling of the interaction of a Cdc25C peptide with Chk1 has uncovered several conserved residues that are important for substrate selectivity. In addition, we found that the less conserved C-terminal region negatively impacts Chk1 kinase activity

Xist recruits the X chromosome to the nuclear lamina to enable chromosome-wide silencing

The Xist long noncoding RNA orchestrates X chromosome inactivation, a process that entails chromosome-wide silencing and remodeling of the three-dimensional (3D) structure of the X chromosome. Yet, it remains unclear whether these changes in nuclear structure are mediated by Xist and whether they are required for silencing. Here, we show that Xist directly interacts with the Lamin B receptor, an integral component of the nuclear lamina, and that this interaction is required for Xist-mediated silencing by recruiting the inactive X to the nuclear lamina and by doing so enables Xist to spread to actively transcribed genes across the X. Our results demonstrate that lamina recruitment changes the 3D structure of DNA, enabling Xist and its silencing proteins to spread across the X to silence transcription

Effect of Porosity Gradient in Gas Diffusion Layer on Cell Performance with Thin-Film Agglomerate Model in Cathode Catalyst Layer of a PEM Fuel Cell

ABSTRACT A one-dimensional, steady-state, two-phase, isothermal numerical simulations were performed to investigate the effect on cell performance of a PEM fuel cell under non-uniform porosity of gas diffusion layer. In the simulation, the non-uniform porosity of gas diffusion layer was taken into account to analyze the transport phenomena of water flooding and mass transport in the gas diffusion layer. The porosity of the gas diffusion layer is treated as a linear function. Furthermore, the structure of the catalyst layer is considered to be a cylindrical thin-film agglomerate. Regarding the distribution analysis of liquid water saturation, oxygen concentration and water concentration depend on the porosity of gas diffusion layer. In the simulation, the ε CG and ε GC represent the porosity of the interfaces between the channel and gas diffusion layer and the gas diffusion layer and the catalyst layer, respectively. The simulation results indicate that when the (ε CG , ε GC ) = (0.8, 0.4), higher liquid water saturation appears in the gas diffusion layer and the catalyst layer. On the contrary, when the (ε CG , ε GC ) = (0.4, 0.4), lower liquid water saturation appears. Once the liquid water produced by the electrochemical reaction and condensate of vapor water may accumulate in the open pores of the gas diffusion layer and reduced the oxygen transport to the catalyst sites. This research attempts to use a thin-film agglomerate model, which analyze the significant transport phenomena of water flooding and mass transport under linear porosity gradient of gas diffusion layer in the cathode of a PEM fuel cell

Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing

Understanding the structure-function relationships at cellular, circuit, and organ-wide scale requires 3D anatomical and phenotypical maps, currently unavailable for many organs across species. At the root of this knowledge gap is the absence of a method that enables whole-organ imaging. Herein, we present techniques for tissue clearing in which whole organs and bodies are rendered macromolecule-permeable and optically transparent, thereby exposing their cellular structure with intact connectivity. We describe PACT (passive clarity technique), a protocol for passive tissue clearing and immunostaining of intact organs; RIMS (refractive index matching solution), a mounting media for imaging thick tissue; and PARS (perfusion-assisted agent release in situ), a method for whole-body clearing and immunolabeling. We show that in rodents PACT, RIMS, and PARS are compatible with endogenous-fluorescence, immunohistochemistry, RNA single-molecule FISH, long-term storage, and microscopy with cellular and subcellular resolution. These methods are applicable for high-resolution, high-content mapping and phenotyping of normal and pathological elements within intact organs and bodies

Phosphorylation of Synaptojanin Differentially Regulates Endocytosis of Functionally Distinct Synaptic Vesicle Pools

The rapid replenishment of synaptic vesicles through endocytosis is crucial for sustaining synaptic transmission during intense neuronal activity. Synaptojanin (Synj), a phosphoinositide phosphatase, is known to play an important role in vesicle recycling by promoting the uncoating of clathrin following synaptic vesicle uptake. Synj has been shown to be a substrate of the minibrain (Mnb) kinase, a fly homolog of the dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A); however, the functional impacts of Synj phosphorylation by Mnb are not well understood. Here we identify that Mnb phosphorylates Synj at S1029 in Drosophila. We find that phosphorylation of Synj at S1029 enhances Synj phosphatase activity, alters interaction between Synj and endophilin, and promotes efficient endocytosis of the active cycling vesicle pool (also referred to as exo-endo cycling pool) at the expense of reserve pool vesicle endocytosis. Dephosphorylated Synj, on the other hand, is deficient in the endocytosis of the active recycling pool vesicles but maintains reserve pool vesicle endocytosis to restore total vesicle pool size and sustain synaptic transmission. Together, our findings reveal a novel role for Synj in modulating reserve pool vesicle endocytosis and further indicate that dynamic phosphorylation and dephosphorylation of Synj differentially maintain endocytosis of distinct functional synaptic vesicle pools. SIGNIFICANCE STATEMENT Synaptic vesicle endocytosis sustains communication between neurons during a wide range of neuronal activities by recycling used vesicle membrane and protein components. Here we identify that Synaptojanin, a protein with a known role in synaptic vesicle endocytosis, is phosphorylated at S1029 in vivo by the Minibrain kinase. We further demonstrate that the phosphorylation status of Synaptojanin at S1029 differentially regulates its participation in the recycling of distinct synaptic vesicle pools. Our results reveal a new role for Synaptojanin in maintaining synaptic vesicle pool size and in reserve vesicle endocytosis. As Synaptojanin and Minibrain perturbations are associated with various neurological disorders, such as Parkinson's, autism, and Down syndrome, understanding mechanisms modulating Synaptojanin function provides valuable insights into processes affecting neuronal communication

Safety and efficacy of intracoronary artery administration of human bone marrow-derived mesenchymal stem cells in STEMI of Lee-Sung pigs—A preclinical study for supporting the feasibility of the OmniMSC-AMI phase I clinical trial

BackgroundThis study tested whether early left intracoronary arterial (LAD) administration of human bone marrow-derived mesenchymal stem cells (hBMMSCs, called OmniMSCs) in acute ST-segment elevation myocardial infarction (STEMI) of Lee-Sung pigs induced by 90 min balloon-occluded LAD was safe and effective.Methods and resultsYoung male Lee-Sung pigs were categorized into SC (sham-operated control, n = 3), AMI-B (STEMI + buffer/21 cc/administered at 90 min after STEMI, n = 6), and AMI-M [acute myocardial infarction (AMI) + hBMMSCs/1.5 × 107/administered at 90 min after STEMI, n = 6] groups. By 2 and 5 months after STEMI, the cardiac magnetic resonance imaging demonstrated that the muscle scar score (MSS) and abnormal cardiac muscle exercise score in the infarct region were significantly increased in the AMI-B than in the SC group that were significantly reversed in the AMI-M group, whereas the left ventricular ejection function by each month (from 1 to 5) displayed an opposite pattern of MSS among the groups (all p &lt; 0.001). By 5 months, histopathological findings of infarct and fibrosis areas and isolectin-B4 exhibited an identical pattern, whereas the cellular expressions of troponin-I/troponin-T/von Willebrand factor exhibited an opposite pattern of MSS among the groups (all p &lt; 0.001). The ST-segment resolution (&gt;80%) was significantly earlier (estimated after 6-h AMI) in the AMI-M group than in the AMI-B group (p &lt; 0.001). The protein expressions of inflammation (IL-1β/TNF-α/NF-κB)/oxidative stress (NOX-1/NOX-2/oxidized protein)/apoptosis (cleaved caspase-3/cleaved PARP)/DNA damage (γ-H2AX) displayed an identical pattern to MSS among the groups, whereas the protein expressions of angiogenesis factors (SDF-1α/VEGF) were significantly and progressively increased from SC, AMI-B, to AMI-M groups (all p &lt; 0.001).ConclusionEarly intra-LAD transfusion of OmniMSC treatment effectively reduced the infarct size and preserved LV function in porcine STEMI