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Recently, we reported that device performance degradation mechanisms, which are generated by the γ-ray irradiation in GaN-based metal-insulator-semiconductor high electron mobility transistors (MIS-HEMTs), use extremely thin gate insulators. When the γ-ray was radiated, the total ionizing dose (TID) effects were generated and the device performance deteriorated. In this work, we investigated the device property alteration and its mechanisms, which were caused by the proton irradiation in GaN-based MIS-HEMTs for the 5 nm-thick Si3N4 and HfO2 gate insulator. The device property, such as threshold voltage, drain current, and transconductance varied by the proton irradiation. When the 5 nm-thick HfO2 layer was employed for the gate insulator, the threshold voltage shift was larger than that of the 5 nm-thick Si3N4 gate insulator, despite the HfO2 gate insulator exhibiting better radiation resistance compared to the Si3N4 gate insulator. On the other hand, the drain current and transconductance degradation were less for the 5 nm-thick HfO2 gate insulator. Unlike the γ-ray irradiation, our systematic research included pulse-mode stress measurements and carrier mobility extraction and revealed that the TID and displacement damage (DD) effects were simultaneously generated by the proton irradiation in GaN-based MIS-HEMTs. The degree of the device property alteration was determined by the competition or superposition of the TID and DD effects for the threshold voltage shift and drain current and transconductance deterioration, respectively. The device property alteration was diminished due to the reduction of the linear energy transfer with increasing irradiated proton energy. We also studied the frequency performance degradation that corresponded to the irradiated proton energy in GaN-based MIS-HEMTs using an extremely thin gate insulator.
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While two-dimensional (2D) hexagonal boron nitride (h-BN) is emerging as an atomically thin and dangling bond-free insulating layer for next-generation electronics and optoelectronics, its practical implementation into miniaturized integrated circuits has been significantly limited due to difficulties in large-scale growth directly on epitaxial semiconductor wafers. Herein, the realization of a wafer-scale h-BN van der Waals heterostructure with a 2 in. AlGaN/GaN high-electron mobility transistor (HEMT) wafer using metal-organic chemical vapor deposition is presented. The combination of state-of-the-art microscopic and spectroscopic analyses and theoretical calculations reveals that the heterointerface between â¼2.5 nm-thick h-BN and AlGaN layers is atomically sharp and exhibits a very weak van der Waals interaction without formation of a ternary or quaternary alloy that can induce undesired degradation of device performance. The fabricated AlGaN/GaN HEMT with h-BN shows very promising performance including a cutoff frequency (fT) and maximum oscillation frequency (fMAX) as high as 28 and 88 GHz, respectively, enabled by an effective passivation of surface defects on the HEMT wafer to deliver accurate information with minimized power loss. These findings pave the way for practical implementation of 2D materials integrated with conventional microelectronic devices and the realization of future all-2D electronics.
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Devices based on AlGaN/GaN heterostructures, for example, Schottky barrier diodes (SBDs) and high electron mobility transistors (HEMTs), have been intensively investigated for applications to high-frequency and high-power areas. Presently, the substrates widely distributed are AlGaN/GaN on SiC for its high performance in radio frequency (RF) applications, for examples high cutoff frequency (fT) or high maximum oscillation frequency (fmax), and AlGaN/GaN on Si for its high power performance, for examples high breakdown voltage or high voltage operation. Chemical vapor deposition (CVD) diamond substrates have a thermal conductivity of 12 W/cm·K, and this is a remarkable point because HEMTs or SBDs on AlGaN/GaN on CVD diamonds are one of the promising alternatives for power and RF applications. In comparison, the thermal conductivity of AlGaN/GaN on a sapphire substrate is 0.33 W/cm·K while that of AlGaN/GaN on a Si substrate is 1.3 W/cm·K and that of AlGaN/GaN on a SiC substrate is 4.9 W/cm·K. In this work, we fabricated SBDs with a 137 mm Schottky channel length on AlGaN/GaN on Si and also on a CVD diamond substrate. We also compared the thermal behaviors of these fabricated large scale SBDs on Si and a CVD diamond substrate.
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The device performance deterioration mechanism caused by the total ionizing dose effect after the γ-ray irradiation was investigated in GaN-based metal-insulator-semiconductor high electron mobility transistors (MIS-HEMTs) for a 5 nm-thick SiN and HfO2 gate dielectric layer. The γ-ray radiation hardness according to the gate dielectric layer was also compared between the two different GaN-based MIS-HEMTs. Although HfO2 has exhibited strong tolerance to the total ionizing dose effect in Si-based devices, there is no detail report of the γ-ray radiation effects in GaN-based MIS-HEMTs employing a HfO2 gate dielectric layer. The pulsed-mode stress measurement results and carrier mobility behavior revealed that the device properties not only have direct current (DC) characteristics, but radio frequency (RF) performance has also been mostly degraded by the deterioration of the gate dielectric quality and the trapped charges inside the gate insulator. We also figured out that the immunity to the γ-ray radiation was improved when HfO2 was employed instead of SiN as a gate dielectric layer due to its stronger endurance to the γ-ray irradiation. Our results highlight that the application of a gate insulator that shows superior immunity to the γ-ray irradiation is a crucial factor for the improvement of the total ionizing dose effect in GaN-based MIS-HEMTs.
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An enhancement-mode AlGaN/GaN metal-insulator-semiconductor high-electron- mobility-transistor was fabricated using a recess gate and CF4 plasma treatment to investigate its reliable applicability to high-power devices and circuits. The fluorinated-gate device showed hysteresis during the DC current-voltage measurement, and the polarity and magnitude of hysteresis depend on the drain voltage. The hysteresis phenomenon is due to the electron trapping at the Al2O3/AlGaN interface and charging times longer than milliseconds were obtained by pulse I-V measurement. In addition, the subthreshold slope of the fluorinated-gate device was increased after the positive gate bias stress because of the two-dimensional electron gas reduction by ionized fluorine. Our systematic observation revealed that the effect of fluorine ions should be considered for the design of AlGaN/GaN power circuits.
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Investigation of the dielectric properties of cell membranes plays an important role in understanding the biological activities that sustain cellular life and realize cellular functionalities. Herein, the variable dielectric polarization characteristics of cell membranes are reported. In controlling the dielectric polarization of a cell using dielectrophoresis force spectroscopy, different cellular crossover frequencies were observed by modulating both the direction and sweep rate of the frequency. The crossover frequencies were used for the extraction of the variable capacitance, which is involved in the dielectric polarization across the cell membranes. In addition, this variable phenomenon was investigated by examining cells whose membranes were cholesterol-depleted with methyl-ß-cyclodextrin, which verified a strong correlation between the variable dielectric polarization characteristics and membrane composition changes. This study presented the dielectric polarization properties in live cells' membranes that can be modified by the regulation of external stimuli and provided a powerful platform to explore cellular membrane dielectric polarization.
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Membrana Celular/metabolismo , Membrana Celular/efectos de los fármacos , Supervivencia Celular , Impedancia Eléctrica , Humanos , Células MCF-7 , beta-Ciclodextrinas/farmacologíaRESUMEN
Fabrication of normally-off field effect transistors (FETs) possessed uniform turn-on threshold voltage (Vth) is of special interests. In this work, they were fabricated using dry etching recess techniques under the gate region, with dry etching conditions of extremely low rate. We report how the recess depth under the gate area induced the Vth shift of normally-off FETs on AlGaN/GaN heterostructure, which were fabricated with a 1.5 nm/min etching rate. Chlorine-based inductively coupled plasma (ICP) was applied to perform the etching process for the AlGaN/GaN heterostructure. Devices were fabricated with different recess depths under the gate area, and examined to determine their performances, particularly the dependence of recess time and recess depth on Vth shift. The applied dry etching conditions resulted in a low-damaged and not-rough morphology on the etched surfaces of AlGaN/GaN. Fine controlled and well defined recess depth of the AlGaN/GaN heterostructure under the gate region was achieved with no etch-stop layers. Conventional fabrication processes were applied with the dry etching conditions of extremely low rate to fabricate normally-off MOSFETs of Al2O3/AlGaN/GaN. The achieved Vth of +5.64 V was high positive and the leakage current of off-state was measured as ~10-6 A/mm.
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High electron mobility transistors (HEMTs) and Schottky barrier diodes (SBDs) based on AlGaN/GaN heterostructure have been widely studied for high-frequency and/or high-power application. Widely distributed substrates for the high performance of RF applications are presently AlGaN/GaN on SiC, and those for high power performance are AlGaN/GaN on Si. Because the thermal conductivity of CVD diamond substrates is as high as 12 W/cm · K, devices on AlGaN/GaN on CVD diamond are one of the excellent alternatives for power and RF applications. In comparison, the thermal conductivity of AlGaN/GaN on SiC is 4.9 W/cm K, and that of AlGaN/GaN on Si is 1.3 W/cm · K. In this work, we report the fabrication of SBD devices with 163.8 mm Schottky channel length. We also compared the thermal properties of the fabricated large scale SBD devices on different substrates.
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We investigate DC characteristics of AlGaN/GaN high-electron mobility transistors by using a source-bridged field plate and additional bottom plate (BP) structure. The analysis of experimental data was performed with a two-dimensional simulator. Source connected BP structure stabilized threshold voltage and transconductance regardless of various drain voltages. The effect of BP location was also analyzed, which had optimal DC values because of the dependence of breakdown voltage and drain current of the device on BP position between gate and drain. Finally, the optimum distance of 0.8 µm from drain side gate head edge to BP was achieved for optimum DC characteristics and the highest breakdown voltage of 341 V.
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In this study, we investigated the operational characteristics of AlGaN/GaN high electron mobility transistors (HEMTs) by applying the copper-filled trench and via structures for improved heat dissipation. Therefore, we used a basic T-gate HEMT device to construct the thermal structures. To identify the heat flow across the device structure, a thermal conductivity model and the heat transfer properties corresponding to the GaN, SiC, and Cu materials were applied. Initially, we simulated the direct current (DC) characteristics of a basic GaN on SiC HEMT to confirm the self-heating effect on AlGaN/GaN HEMT. Then, to verify the heat sink effect of the copper-filled thermal structures, we compared the DC characteristics such as the threshold voltage, transconductance, saturation current, and breakdown voltage. Finally, we estimated and compared the lattice temperature of a two-dimensional electron gas channel, the vertical lattice temperature near the drain-side gate head edge, and the transient thermal analysis for the copper-filled thermal trench and via structures. Through this study, we could optimize the operational characteristics of the device by applying an effective heat dissipation structure to the AlGaN/GaN HEMT.
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PURPOSE: There is still a clinical need to easily evaluate the metastatic status of lymph nodes during breast cancer surgery. We hypothesized that ex vivo shear-wave elastography (SWE) would predict precisely the presence of metastasis in the excised lymph nodes. METHODS: A total of 63 patients who underwent breast cancer surgery were prospectively enrolled in this study from May 2014 to April 2015. The excised axillary lymph nodes were examined using ex vivo SWE. Metastatic status was confirmed based on the final histopathological diagnosis of the permanent section. Lymph node characteristics and elasticity values measured by ex vivo SWE were assessed for possible association with nodal metastasis. RESULTS: A total of 274 lymph nodes, harvested from 63 patients, were examined using ex vivo SWE. The data obtained from 228 of these nodes from 55 patients were included in the analysis. Results showed that 187 lymph nodes (82.0%) were nonmetastatic and 41 lymph nodes (18.0%) were metastatic. There was significant difference between metastatic and nonmetastatic nodes with respect to the mean (45.4 kPa and 17.7 kPa, p<0.001) and maximum (55.3 kPa and 23.2 kPa, p<0.001) stiffness. The elasticity ratio was higher in the metastatic nodes (4.36 and 1.57, p<0.001). Metastatic nodes were significantly larger than nonmetastatic nodes (mean size, 10.5 mm and 7.5 mm, p<0.001). The size of metastatic nodes and nodal stiffness were correlated (correlation coefficient of mean stiffness, r=0.553). The area under curve of mean stiffness, maximum stiffness, and elasticity ratio were 0.794, 0.802, and 0.831, respectively. CONCLUSION: Ex vivo SWE may be a feasible method to predict axillary lymph node metastasis intraoperatively in patients undergoing breast cancer surgery.
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Facioscapulohumeral muscular dystrophy (FSHD) is caused by insufficient epigenetic repression of D4Z4 macrosatellite repeat where DUX4, an FSHD causing gene is embedded. There are two forms of FSHD, FSHD1 with contraction of D4Z4 repeat and FSHD2 with chromatin compaction defects mostly due to SMCHD1 mutation. Previous reports showed DUX4-induced gene expression changes as well as changes in microRNA expression in FSHD muscle cells. However, a genome wide analysis of small noncoding RNAs that might be regulated by DUX4 or by mutations in SMCHD1 has not been reported yet. Here, we identified several types of small noncoding RNAs including known microRNAs that are differentially expressed in FSHD2 muscle cells compared to control. Although fewer small RNAs were differentially expressed during muscle differentiation in FSHD2 cells compared to controls, most of the known myogenic microRNAs, such as miR1, miR133a and miR206 were induced in both FSHD2 and control muscle cells during differentiation. Our small RNA sequencing data analysis also revealed both DUX4- and SMCHD1-specific changes in FSHD2 muscle cells. Six FSHD2 microRNAs were affected by DUX4 overexpression in control myoblasts, whereas increased expression of tRNAs and 5S rRNAs in FSHD2 muscle cells was largely recapitulated in SMCHD1-depleted control myoblasts. Altogether, our studies suggest that the small noncoding RNA transcriptome changes in FSHD2 might be different from those in FSHD1 and that these differences may provide new diagnostic and therapeutic tools specific to FSHD2.
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Proteínas Cromosómicas no Histona/genética , Proteínas de Homeodominio/genética , Distrofia Muscular Facioescapulohumeral/genética , ARN Pequeño no Traducido/genética , Estudios de Casos y Controles , Diferenciación Celular/genética , Regulación de la Expresión Génica , Humanos , MicroARNs/genética , Fibras Musculares Esqueléticas/patología , Fibras Musculares Esqueléticas/fisiología , Mutación , Mioblastos/patología , Mioblastos/fisiología , ARN Ribosómico 5S/genética , ARN de Transferencia/genética , Reproducibilidad de los ResultadosRESUMEN
The DUX4 transcription factor is encoded by a retrogene embedded in each unit of the D4Z4 macrosatellite repeat. DUX4 is normally expressed in the cleavage-stage embryo, whereas chromatin repression prevents DUX4 expression in most somatic tissues. Failure of this repression causes facioscapulohumeral muscular dystrophy (FSHD) due to mis-expression of DUX4 in skeletal muscle. In this study, we used CRISPR/Cas9 engineered chromatin immunoprecipitation (enChIP) locus-specific proteomics to characterize D4Z4-associated proteins. These and other approaches identified the Nucleosome Remodeling Deacetylase (NuRD) and Chromatin Assembly Factor 1 (CAF-1) complexes as necessary for DUX4 repression in human skeletal muscle cells and induced pluripotent stem (iPS) cells. Furthermore, DUX4-induced expression of MBD3L proteins partly relieved this repression in FSHD muscle cells. Together, these findings identify NuRD and CAF-1 as mediators of DUX4 chromatin repression and suggest a mechanism for the amplification of DUX4 expression in FSHD muscle cells.
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Epigénesis Genética , Proteínas de Homeodominio/genética , Distrofia Muscular Facioescapulohumeral/genética , Cromatina/genética , Factor 1 de Ensamblaje de la Cromatina/genética , Repeticiones Palindrómicas Cortas Agrupadas y Regularmente Espaciadas , Regulación del Desarrollo de la Expresión Génica , Silenciador del Gen , Proteínas de Homeodominio/química , Humanos , Complejo Desacetilasa y Remodelación del Nucleosoma Mi-2/química , Complejo Desacetilasa y Remodelación del Nucleosoma Mi-2/genética , Fibras Musculares Esqueléticas/química , Fibras Musculares Esqueléticas/patología , Músculo Esquelético/química , Músculo Esquelético/metabolismo , Distrofia Muscular Facioescapulohumeral/fisiopatología , Nucleosomas/química , Nucleosomas/genéticaRESUMEN
Magnetic nanoparticles (MNPs) are widely used in biomedical and clinical applications, including medical imaging, therapeutics, and biological sample processing. Rapid characterization of MNPs, notably their magnetic moments, should facilitate optimization of particle synthesis and accelerate assay development. Here, we report a compact and low-cost magnetometer for fast, on-site MNP characterization. Termed integrated microHall magnetometer (iHM), our device was fabricated using standard semiconductor processes: an array of Hall sensors, transistor switches, and amplifiers were integrated into a single chip, thus improving the detection sensitivity and facilitating chip operation. By applying the iHM, we demonstrate versatile magnetic assays. We measured the magnetic susceptibility and moments of MNPs using small sample amounts (â¼10 pL), identified different MNP compositions in mixtures, and detected MNP-labeled single cells.
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Dispositivos Laboratorio en un Chip , Nanopartículas de Magnetita/química , Magnetometría/instrumentación , Línea Celular Tumoral , Diseño de Equipo , Humanos , Nanopartículas de Magnetita/análisis , Magnetometría/métodos , Metales/químicaRESUMEN
BACKGROUND: Facioscapulohumeral muscular dystrophy (FSHD) is in most cases caused by a contraction of the D4Z4 macrosatellite repeat on chromosome 4 (FSHD1) or by mutations in the SMCHD1 or DNMT3B gene (FSHD2). Both situations result in the incomplete epigenetic repression of the D4Z4-encoded retrogene DUX4 in somatic cells, leading to the aberrant expression of DUX4 in the skeletal muscle. In mice, Smchd1 regulates chromatin repression at different loci, having a role in CpG methylation establishment and/or maintenance. METHODS: To investigate the global effects of harboring heterozygous SMCHD1 mutations on DNA methylation in humans, we combined 450k methylation analysis on mononuclear monocytes from female heterozygous SMCHD1 mutation carriers and unaffected controls with reduced representation bisulfite sequencing (RRBS) on FSHD2 and control myoblast cell lines. Candidate loci were then evaluated for SMCHD1 binding using ChIP-qPCR and expression was evaluated using RT-qPCR. RESULTS: We identified a limited number of clustered autosomal loci with CpG hypomethylation in SMCHD1 mutation carriers: the protocadherin (PCDH) cluster on chromosome 5, the transfer RNA (tRNA) and 5S rRNA clusters on chromosome 1, the HOXB and HOXD clusters on chromosomes 17 and 2, respectively, and the D4Z4 repeats on chromosomes 4 and 10. Furthermore, minor increases in RNA expression were seen in FSHD2 myoblasts for some of the PCDHß cluster isoforms, tRNA isoforms, and a HOXB isoform in comparison to controls, in addition to the previously reported effects on DUX4 expression. SMCHD1 was bound at DNAseI hypersensitivity sites known to regulate the PCDHß cluster and at the chromosome 1 tRNA cluster, with decreased binding in SMCHD1 mutation carriers at the PCDHß cluster sites. CONCLUSIONS: Our study is the first to investigate the global methylation effects in humans resulting from heterozygous mutations in SMCHD1. Our results suggest that SMCHD1 acts as a repressor on a limited set of autosomal gene clusters, as an observed reduction in methylation associates with a loss of SMCHD1 binding and increased expression for some of the loci.
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Proteínas Cromosómicas no Histona/genética , Metilación de ADN , Sitios Genéticos , Distrofia Muscular Facioescapulohumeral/genética , Células Cultivadas , Proteínas Cromosómicas no Histona/metabolismo , Islas de CpG , Femenino , Heterocigoto , Humanos , Familia de Multigenes , Distrofia Muscular Facioescapulohumeral/metabolismo , Mutación , Mioblastos/metabolismo , Unión ProteicaRESUMEN
To better understand transcriptional regulation during human oogenesis and preimplantation development, we defined stage-specific transcription, which highlighted the cleavage stage as being highly distinctive. Here, we present multiple lines of evidence that a eutherian-specific multicopy retrogene, DUX4, encodes a transcription factor that activates hundreds of endogenous genes (for example, ZSCAN4, KDM4E and PRAMEF-family genes) and retroviral elements (MERVL/HERVL family) that define the cleavage-specific transcriptional programs in humans and mice. Remarkably, mouse Dux expression is both necessary and sufficient to convert mouse embryonic stem cells (mESCs) into 2-cell-embryo-like ('2C-like') cells, measured here by the reactivation of '2C' genes and repeat elements, the loss of POU5F1 (also known as OCT4) protein and chromocenters, and the conversion of the chromatin landscape (as assessed by transposase-accessible chromatin using sequencing (ATAC-seq)) to a state strongly resembling that of mouse 2C embryos. Thus, we propose mouse DUX and human DUX4 as major drivers of the cleavage or 2C state.
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Proteínas de Homeodominio/metabolismo , Retroelementos/genética , Adulto , Empalme Alternativo , Animales , Blastocisto/fisiología , Cromatina/genética , Cromatina/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Proteínas de Homeodominio/genética , Humanos , Ratones Transgénicos , Oocitos/fisiología , TranscriptomaRESUMEN
Facioscapulohumeral muscular dystrophy (FSHD) is caused by the aberrant expression of the DUX4 transcription factor in skeletal muscle. The DUX4 retrogene is encoded in the D4Z4 macrosatellite repeat array, and smaller array size or a mutation in the SMCHD1 gene results in inefficient epigenetic repression of DUX4 in skeletal muscle, causing FSHD1 and FSHD2, respectively. Previously we showed that the entire D4Z4 repeat is bi-directionally transcribed with the generation of small si- or miRNA-like fragments and suggested that these might suppress DUX4 expression through the endogenous RNAi pathway. Here we show that exogenous siRNA targeting the region upstream of the DUX4 transcription start site suppressed DUX4 mRNA expression and increased both H3K9 methylation and AGO2 recruitment. In contrast, similarly targeted MOE-gapmer antisense oligonucleotides that degrade RNA but do not engage the RNAi pathway did not repress DUX4 expression. In addition, knockdown of DICER or AGO2 using either siRNA or MOE-gapmer chemistries resulted in the induction of DUX4 expression in control muscle cells that normally do not express DUX4, indicating that the endogenous RNAi pathway is necessary to maintain repression of DUX4 in control muscle cells. Together these data demonstrate a role of the endogenous RNAi pathway in repeat-mediated epigenetic repression of the D4Z4 macrosatellite repeat, and show that enhancing the activity of this pathway by supplying exogenous siRNA oligonucleotides represents a potential therapeutic approach to silencing DUX4 in FSHD.
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Proteínas Argonautas/metabolismo , Epigénesis Genética , Silenciador del Gen , Repeticiones de Microsatélite , Distrofia Muscular Facioescapulohumeral/genética , Distrofia Muscular Facioescapulohumeral/metabolismo , ARN Interferente Pequeño/genética , Ribonucleasa III/metabolismo , Línea Celular , Cromatina/genética , Cromatina/metabolismo , Regulación de la Expresión Génica , Histonas/metabolismo , Proteínas de Homeodominio/genética , Humanos , Distrofia Muscular Facioescapulohumeral/terapia , Interferencia de ARN , Sitio de Iniciación de la TranscripciónRESUMEN
Skeletal muscle development is orchestrated by the myogenic regulatory factor MyoD, whose activity is blocked in myoblasts by proteins preventing its nuclear translocation and/or binding to G/C-centered E-boxes in target genes. Recent evidence indicates that muscle gene expression is also regulated at the cis level by differential affinity for DNA between MyoD and other E-box binding proteins during myogenesis. MyoD binds to G/C-centered E-boxes, enriched in muscle differentiation genes, in myotubes but not in myoblasts. Here, we used cell-based and in vivo Drosophila, Xenopus laevis, and mouse models to show that ZEB1, a G/C-centered E-box binding transcriptional repressor, imposes a temporary stage-dependent inhibition of muscle gene expression and differentiation via CtBP-mediated transcriptional repression. We found that, contrary to MyoD, ZEB1 binds to G/C-centered E-boxes in muscle differentiation genes at the myoblast stage but not in myotubes. Its knockdown results in precocious expression of muscle differentiation genes and acceleration of myotube formation. Inhibition of muscle genes by ZEB1 occurs via transcriptional repression and involves recruitment of the CtBP corepressor. Lastly, we show that the pattern of gene expression associated with muscle differentiation is accelerated in ZEB1(-/-) mouse embryos. These results set ZEB1 as an important regulator of the temporal pattern of gene expression controlling muscle differentiation.
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Diferenciación Celular/genética , Regulación de la Expresión Génica , Proteínas de Homeodominio/genética , Factores de Transcripción de Tipo Kruppel/genética , Músculo Esquelético/fisiología , Activación Transcripcional , Oxidorreductasas de Alcohol/genética , Oxidorreductasas de Alcohol/metabolismo , Animales , Línea Celular , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Elementos E-Box , Expresión Génica , Proteínas de Homeodominio/metabolismo , Factores de Transcripción de Tipo Kruppel/metabolismo , Ratones , Ratones Endogámicos C57BL , Desarrollo de Músculos/genética , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/citología , Músculo Esquelético/metabolismo , Proteína MioD/genética , Proteína MioD/metabolismo , Mioblastos/metabolismo , Proteínas Represoras/genética , Proteínas Represoras/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Transcripción Genética , Xenopus laevis , Homeobox 1 de Unión a la E-Box con Dedos de ZincRESUMEN
Regulating the transition from lineage-restricted progenitors to terminally differentiated cells is a central aspect of nervous system development. Here, we investigated the role of the nucleoprotein geminin in regulating neurogenesis at a mechanistic level during both Xenopus primary neurogenesis and mammalian neuronal differentiation in vitro. The latter work utilized neural cells derived from embryonic stem and embryonal carcinoma cells in vitro and neural stem cells from mouse forebrain. In all of these contexts, geminin antagonized the ability of neural basic helix-loop-helix (bHLH) transcription factors to activate transcriptional programs promoting neurogenesis. Furthermore, geminin promoted a bivalent chromatin state, characterized by the presence of both activating and repressive histone modifications, at genes encoding transcription factors that promote neurogenesis. This epigenetic state restrains the expression of genes that regulate commitment of undifferentiated stem and neuronal precursor cells to neuronal lineages. However, maintaining geminin at high levels was not sufficient to prevent terminal neuronal differentiation. Therefore, these data support a model whereby geminin promotes the neuronal precursor cell state by modulating both the epigenetic status and expression of genes encoding neurogenesis-promoting factors. Additional developmental signals acting in these cells can then control their transition toward terminal neuronal or glial differentiation during mammalian neurogenesis.
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Proteínas de Ciclo Celular/genética , Epigénesis Genética , Neurogénesis/genética , Proteínas Nucleares/genética , Prosencéfalo/metabolismo , Xenopus laevis/genética , Animales , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Proteínas de Ciclo Celular/metabolismo , Diferenciación Celular , Cromatina/genética , Cromatina/metabolismo , Embrión no Mamífero , Células Madre Embrionarias/citología , Células Madre Embrionarias/metabolismo , Geminina , Regulación del Desarrollo de la Expresión Génica , Histonas/genética , Histonas/metabolismo , Ratones , Células Madre Neoplásicas/citología , Células Madre Neoplásicas/metabolismo , Células-Madre Neurales/citología , Células-Madre Neurales/metabolismo , Neuronas/citología , Neuronas/metabolismo , Proteínas Nucleares/metabolismo , Prosencéfalo/citología , Prosencéfalo/embriología , Activación Transcripcional , Proteínas de Xenopus , Xenopus laevis/embriología , Xenopus laevis/metabolismoRESUMEN
PURPOSE: Laparoscopic totally extraperitoneal (TEP) herniorrhaphy has been recognized as a treatment option for inguinal hernia. The objective of this study was to clarify the learning curve for laparoscopic TEP herniorrhaphy using the moving average method. METHODS: A total of 90 patients underwent laparoscopic TEP herniorrhaphy by a single surgeon between March 2009 and March 2011. We analyzed medical records including the demographic data, operating time, hospital stay, and postoperative complications. RESULTS: The mean operating time of the initial 30 cases (learning period group) was 66.3 minutes. After the initial 30 cases were performed, the time decreased to 52.8 minutes in the later 60 cases (experienced period group, P = 0.015). This represents the operating time becoming stabilized and then decreasing as the number of performed cases accumulates. Hospital stay was shorter and frequency of pain control, and complication rate were lower in the experienced period, however, there was no statistical significance. CONCLUSION: We suggest that number of patients needed for the learning curve for laparoscopic TEP herniorrhaphy should be 30 cases. The operating time for laparoscopic TEP herniorrhaphy stabilizes after 40 cases in moving average analysis.