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The COVID-19 pandemic has caused a massive disruption in the way traditional higher education institutions deliver their courses. Unlike transitions from face-to-face teaching to blended, online or flipped classroom in the past, changes in emergency remote teaching -a temporary shift of instructional delivery to an alternate remote delivery mode due to crisis circumstances- happen suddenly and in an unplanned way. This study analyzes the move to emergency remote teaching at the School of Telecommunication Engineering (Universidad Politécnica de Madrid), and the impact of organizational aspects related to unplanned change, instruction-related variables -class size, synchronous/asynchronous delivery- and use of digital supporting technologies, on students' academic performance. Using quantitative data of academic records across all (N = 43) courses of a bachelor's degree programme in Telecommunication Engineering and qualitative data from a questionnaire delivered to all (N = 43) course coordinators, the research also compares the academic results of students during the COVID-19 pandemic with those of previous years. The results of this case study show an increase in students' academic performance in emergency remote teaching, and support the idea that organizational factors may contribute to successful implementation of emergency remote teaching; the analysis does not find differences across courses with different class sizes or delivery modes. The study further explores possible explanations for the results of the analysis, considering organizational, individual and instruction-related aspects.
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The effective search for the missing and identification of persons, alive or dead, are core components in the prevention and in resolving the issue of Missing Persons. Despite the growing literature on this topic, there is still a lack of publications describing the Search as a process that includes different phases inherently composed of forensic investigative and identification principles for both living and deceased missing persons. This paper is the result of discussions between the Forensic Unit of the International Committee of the Red Cross (ICRC) and members of its external Forensic Advisory Board. It aims to present the Search process as an overarching concept that includes the investigation and identification phases of the missing in any state (dead or alive), in any scenario (with or without bodies), with an integrated, multidisciplinary, and multiagency approach for implementation by all actors involved in the investigation and identification phases of missing persons.
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The precise control and stabilization of magnetic domain walls is key for the development of the next generation magnetic nano-devices. Among the multitude of magnetic configurations of a magnetic domain wall, topologically protected states are of particular interest due to their intrinsic stability. In this work, using XMCD-PEEM, we have observed a topologically protected magnetic domain wall in a ferromagnetic cylindrical nanowire. Its structure is stabilized by periodic sharp alterations of the chemical composition in the nanowire. The large stability of this topologically protected domain wall contrasts with the mobility of other non-protected and non-chiral states also present in the same nanowire. The micromagnetic simulations show the structure and the conditions required to find the topologically protected state. These results are relevant for the design of future spintronic devices such as domain wall based RF oscillators or magnetic memories.
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The objective of this study was to apply the method for calculating dental age proposed by Demirjian et al. to a sample of Spanish children, followed by a comparison between their dental and chronological ages. This study also set out to create tables to convert specific dental age using the maturity data from our sample. This study was performed on a sample of 1010 orthopantograms taken of Spanish children (485 boys and 525 girls) aged 2-16. We found that the mean estimated dental age exceeded the mean chronological age in both boys and girls, with the mean difference being 0.87 and 0.55 years respectively. We adapted Demirjian's method to our study sample to obtain specific conversion tables and curves.
Assuntos
Determinação da Idade pelos Dentes/métodos , Dente/crescimento & desenvolvimento , Adolescente , Criança , Pré-Escolar , Feminino , Odontologia Legal , Humanos , Masculino , Radiografia Panorâmica , Caracteres Sexuais , Espanha , Dente/diagnóstico por imagemRESUMO
The fundamental process of ribosome biogenesis requires hundreds of factors and takes place in the nucleolus. This process has been most thoroughly characterized in baker's yeast and is generally well conserved from yeast to humans. However, some of the required proteins in yeast are not found in humans, raising the possibility that they have been replaced by functional analogs. Our objective was to identify non-conserved interaction partners for the human ribosome biogenesis factor, hUTP4/Cirhin, since the R565W mutation in the C-terminus of hUTP4/Cirhin was reported to cause North American Indian childhood cirrhosis (NAIC). By screening a yeast two-hybrid cDNA library derived from human liver, and through affinity purification followed by mass spectrometry, we identified an uncharacterized nucleolar protein, NOL11, as an interaction partner for hUTP4/Cirhin. Bioinformatic analysis revealed that NOL11 is conserved throughout metazoans and their immediate ancestors but is not found in any other phylogenetic groups. Co-immunoprecipitation experiments show that NOL11 is a component of the human ribosomal small subunit (SSU) processome. siRNA knockdown of NOL11 revealed that it is involved in the cleavage steps required to generate the mature 18S rRNA and is required for optimal rDNA transcription. Furthermore, abnormal nucleolar morphology results from the absence of NOL11. Finally, yeast two-hybrid analysis shows that NOL11 interacts with the C-terminus of hUTP4/Cirhin and that the R565W mutation partially disrupts this interaction. We have therefore identified NOL11 as a novel protein required for the early stages of ribosome biogenesis in humans. Our results further implicate a role for NOL11 in the pathogenesis of NAIC.
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Indígenas Norte-Americanos/genética , Cirrose Hepática/genética , Proteínas Nucleares/genética , RNA Ribossômico 18S/genética , Ribonucleoproteínas/genética , Ribossomos/genética , Sítios de Ligação , Nucléolo Celular/genética , Nucléolo Celular/metabolismo , Criança , Sequência Conservada , Biblioteca Gênica , Células HeLa , Humanos , Imunoprecipitação , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Mutação , Proteínas Nucleares/antagonistas & inibidores , Proteínas Nucleares/metabolismo , Ligação Proteica , Estrutura Terciária de Proteína , Precursores de RNA , RNA Ribossômico 18S/metabolismo , RNA Interferente Pequeno/genética , Ribonucleoproteínas/metabolismo , Ribossomos/metabolismo , Ribossomos/patologia , Técnicas do Sistema de Duplo-HíbridoAssuntos
Tumores Neuroendócrinos/diagnóstico , Neoplasias Pancreáticas/diagnóstico , Protocolos Clínicos , Endossonografia , Gastrinoma/diagnóstico , Gastrinoma/diagnóstico por imagem , Gastrinoma/patologia , Humanos , Insulinoma/diagnóstico , Insulinoma/diagnóstico por imagem , Insulinoma/patologia , Imageamento por Ressonância Magnética , Tumores Neuroendócrinos/diagnóstico por imagem , Tumores Neuroendócrinos/patologia , Neoplasias Pancreáticas/diagnóstico por imagem , Neoplasias Pancreáticas/patologia , Tomografia por Emissão de Pósitrons , Tomografia Computadorizada por Raios XRESUMO
The purpose of this study is to clarify the chronology of different stages of dental development, according to Demirjian, in a sample of Spanish children, which will enable us to build a database that will be used as a reference in regard to the dental development of individuals of our socio-geographic environment. In the same studied sample, a calculation of the dental age according to Demirjian was carried out. This study was conducted in a final sample consisting of 1010 orthopantograms, corresponding to Spanish children (485 boys and 525 girls) ages 2-16. Comparing the age of onset of the different stages among the children, evidence was found that girls had an earlier general development than boys. These differences were only statistically significant in teeth and concrete stages. The canine teeth revealed greater gender dimorphism, with significant differences in all stages compared with the upper canines. The method proposed by Demirjian for dental age calculation resulted in a significant overestimation of dental age in relation to the chronological age in boys (average of 0.87 years) and girls (average of 0.55 years). Data from this study may be used as reference for dental maturity, as well as a standard for estimating age in Spanish children.
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Determinação da Idade pelos Dentes/métodos , Dentição Permanente , Dente/crescimento & desenvolvimento , Adolescente , Criança , Pré-Escolar , Feminino , Odontologia Legal/métodos , Humanos , Masculino , Radiografia Panorâmica , Caracteres Sexuais , Espanha , Dente/diagnóstico por imagemRESUMO
In mammalian cells, transcriptionally active ribosomal genes are replicated in the early S phase, and the silent ribosomal genes in the late S phase, though mechanisms of this timing remain unknown. UBF (Upstream Binding Factor), a DNA binding protein and component of the pol I transcription machinery, is considered to be responsible for the loose chromatin structure of the active rDNA. Here we question whether such structure alone can ensure early replication of DNA. We investigate this problem on the model of pseudo-NORs, the tandem arrays of heterologous DNA sequence with high affinity for UBF, introduced into human chromosomes. Such arrays are not transcribed, yet efficiently bind UBF and mimic the chromatin structure of active rDNA. In our study, a human derived stable cell line containing one pseudo-NOR on the chromosome 10 was transiently transfected with UBF-GFP and PCNA-RFP, which allowed us to observe in vivo the growth of pseudo-NORs resulted from their replication. We found that replication of pseudo-NORs is not restricted to the early S phase, but continues in the late S phase at a significant level. These results were confirmed in the experiments with incorporation of thymidin analog EdU and BrdU ChIP assay. Similar results were obtained with another cell line containing pseudo-NOR on the chromosome 7. Our data indicate that the specific loose structure of chromatin, produced by the architect protein UBF, is not sufficient for the early replication.
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Região Organizadora do Nucléolo/metabolismo , Linhagem Celular Tumoral , DNA Ribossômico/genética , DNA Ribossômico/metabolismo , Humanos , Imuno-Histoquímica , Região Organizadora do Nucléolo/genética , Proteínas Pol1 do Complexo de Iniciação de Transcrição/genética , Proteínas Pol1 do Complexo de Iniciação de Transcrição/metabolismo , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Fase S/genética , Fase S/fisiologiaRESUMO
Eukaryotic 18S rRNA processing is mediated by the small subunit (SSU) processome, a machine comprised of the U3 small nucleolar RNP (U3 snoRNP), tUTP, bUTP, MPP10, and BMS1/RCL1 subcomplexes. We report that the human SSU processome is a dynamic structure with the recruitment and release of subcomplexes occurring during the early stages of ribosome biogenesis. A novel 50S U3 snoRNP accumulated when either pre-rRNA transcription was blocked or the tUTP proteins were depleted. This complex did not contain the tUTP, bUTP, MPP10, and BMS1/RCL1 subcomplexes but was associated with the RNA-binding proteins nucleolin and RRP5 and the RNA helicase DBP4. Our data suggest that the 50S U3 snoRNP is an SSU assembly intermediate that is likely recruited to the pre-rRNA through the RNA-binding proteins nucleolin and RRP5. We predict that nucleolin is only transiently associated with the SSU processome and likely leaves the complex not long after 50S U3 snoRNP recruitment. The nucleolin-binding site potentially overlaps that of several other key factors, and we propose that this protein must leave the SSU processome for pre-rRNA processing to occur.
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Proteínas Nucleares/metabolismo , Fosfoproteínas/metabolismo , Subunidades Proteicas/metabolismo , RNA Helicases/metabolismo , Proteínas de Ligação a RNA/metabolismo , Ribonucleoproteínas Nucleolares Pequenas/metabolismo , Nucléolo Celular , RNA Helicases DEAD-box , Células HeLa , Humanos , Ligação Proteica , Transporte Proteico , RNA Polimerase I/metabolismo , Precursores de RNA/metabolismo , Transcrição Gênica , NucleolinaRESUMO
Nucleolar organiser regions (NORs) are comprised of tandem arrays of ribosomal gene (rDNA) repeats that are transcribed by RNA polymerase I (Pol I), ultimately resulting in formation of a nucleolus. Upstream binding factor (UBF), a DNA binding protein and component of the Pol I transcription machinery, binds extensively across the rDNA repeat in vivo. Pseudo-NORs are tandem arrays of a heterologous DNA sequence with high affinity for UBF introduced into human chromosomes. In this review we describe how analysis of pseudo-NORs has provided important insights into nucleolar formation. Pseudo-NORs mimic endogenous NORs in a number of important respects. On metaphase chromosomes both appear as secondary constrictions comprised of undercondensed chromatin. The transcriptional silence of pseudo-NORs provides a platform for studying the transcription independent recruitment of factors required for nucleolar formation by this specialised chromatin structure. During interphase, pseudo-NORs appear as distinct and novel sub-nuclear bodies. Analysis of these bodies and comparison to their endogenous counterpart has provided insights into nucleolar formation and structure.
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Nucléolo Celular/metabolismo , DNA Ribossômico/metabolismo , Região Organizadora do Nucléolo/metabolismo , Ciclo Celular/fisiologia , Nucléolo Celular/ultraestrutura , Cromatina/genética , Cromatina/metabolismo , Cromossomos Humanos , DNA Ribossômico/genética , Humanos , Região Organizadora do Nucléolo/ultraestrutura , Proteínas Pol1 do Complexo de Iniciação de Transcrição/genética , Proteínas Pol1 do Complexo de Iniciação de Transcrição/metabolismo , RNA Polimerase I/metabolismo , Transcrição GênicaRESUMO
Efficient ribosome biogenesis requires coordination of a highly complex series of events. Early events include pre-RNA transcription, processing, and modification. Analysis in yeast has demonstrated that t-UTPs, components of the U3 snoRNA-containing pre-rRNA processing complex, are required for efficient transcription of ribosomal genes (rDNA) by RNA polymerase I (pol I). Here, we characterize human t-UTPs and establish that their ability to link transcription and pre-rRNA processing is evolutionarily conserved. The pol I transcription factor UBF binds extensively across rDNA throughout the cell cycle, resulting in a specialized form of chromatin that is the hallmark of active nucleolar organizer regions (NORs). Transcriptionally silent pseudo-NORs are ectopic, chromosomally integrated, artificial arrays that mimic this specialized chromatin structure. Pseudo-NORs sequester t-UTPs and factors linking transcription with pre-rRNA modification (Nopp140 and Treacle). Recruitment is independent of transcription, the underlying DNA sequence, and location within the nucleolus. Previously, we have demonstrated that pseudo-NORs sequester every component of the pol I transcription machinery. Taken together, these results highlight the importance of the specialized chromatin structure at active NORs in coordinating early events in ribosome biogenesis and nucleolar formation.
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Região Organizadora do Nucléolo/metabolismo , Proteínas Pol1 do Complexo de Iniciação de Transcrição/metabolismo , Precursores de RNA/metabolismo , Sequência de Bases , Ciclo Celular , Cromatina/genética , Cromatina/metabolismo , DNA Ribossômico/genética , DNA Ribossômico/metabolismo , Células HeLa , Humanos , Complexos Multiproteicos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Região Organizadora do Nucléolo/genética , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , RNA Polimerase I/genética , RNA Polimerase I/metabolismo , Precursores de RNA/genética , Processamento Pós-Transcricional do RNA , RNA Interferente Pequeno/genética , Ribonucleoproteínas Nucleolares Pequenas/genética , Ribonucleoproteínas Nucleolares Pequenas/metabolismo , Transcrição GênicaRESUMO
Human ribosomal genes are located in NORs (nucleolar organizer regions) on the short arms of acrocentric chromosomes. During metaphase, previously active NORs appear as prominent chromosomal features termed secondary constrictions, which are achromatic in chromosome banding and positive in silver staining. The architectural RNA polymerase I transcription factor UBF (upstream binding factor) binds extensively across the ribosomal gene repeat throughout the cell cycle. Evidence that UBF underpins NOR structure is provided by an examination of cell lines in which large arrays of a heterologous UBF binding sequences are integrated at ectopic sites on human chromosomes. These arrays efficiently recruit UBF even to sites outside the nucleolus, and during metaphase form novel silver-stainable secondary constrictions, termed pseudo-NORs, that are morphologically similar to NORs.
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Cromatina/genética , Cromatina/metabolismo , Região Organizadora do Nucléolo/genética , Região Organizadora do Nucléolo/metabolismo , Proteínas Pol1 do Complexo de Iniciação de Transcrição/genética , Proteínas Pol1 do Complexo de Iniciação de Transcrição/metabolismo , Ribossomos/genética , Ribossomos/metabolismo , Sítios de Ligação/genética , DNA Ribossômico/genética , DNA Ribossômico/metabolismo , Humanos , Ligação ProteicaRESUMO
The light from historical supernovae could in principle still be visible as scattered-light echoes centuries after the explosion. The detection of light echoes could allow us to pinpoint the supernova event both in position and age and, most importantly, permit the acquisition of spectra to determine the 'type' of the supernova centuries after the direct light from the explosion first reached Earth. Although echoes have been discovered around some nearby extragalactic supernovae, targeted searches have not found any echoes in the regions of historical Galactic supernovae. Here we report three faint variable-surface-brightness complexes with high apparent proper motions pointing back to three of the six smallest (and probably youngest) previously catalogued supernova remnants in the Large Magellanic Cloud, which are believed to have been thermonuclear (type Ia) supernovae. Using the distance and apparent proper motions of these echo arcs, we estimate ages of 610 and 410 years for two of them.
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Human ribosomal genes (rDNA) are located in nucleolar organizer regions (NORs) on the short arms of acrocentric chromosomes. Metaphase NORs that were transcriptionally active in the previous cell cycle appear as prominent chromosomal features termed secondary constrictions that are achromatic in chromosome banding and positive in silver staining. The architectural RNA polymerase I (pol I) transcription factor UBF binds extensively across rDNA throughout the cell cycle. To determine if UBF binding underpins NOR structure, we integrated large arrays of heterologous UBF-binding sequences at ectopic sites on human chromosomes. These arrays efficiently recruit UBF even to sites outside the nucleolus and, during metaphase, form novel silver stainable secondary constrictions, termed pseudo-NORs, morphologically similar to NORs. We demonstrate for the first time that in addition to UBF the other components of the pol I machinery are found associated with sequences across the entire human rDNA repeat. Remarkably, a significant fraction of these same pol I factors are sequestered by pseudo-NORs independent of both transcription and nucleoli. Because of the heterologous nature of the sequence employed, we infer that sequestration is mediated primarily by protein-protein interactions with UBF. These results suggest that extensive binding of UBF is responsible for formation and maintenance of the secondary constriction at active NORs. Furthermore, we propose that UBF mediates recruitment of the pol I machinery to nucleoli independently of promoter elements.
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Região Organizadora do Nucléolo/ultraestrutura , Proteínas Pol1 do Complexo de Iniciação de Transcrição/metabolismo , RNA Polimerase I/metabolismo , Transcrição Gênica , Animais , Sítios de Ligação/genética , Linhagem Celular , Nucléolo Celular/metabolismo , Cromossomos , DNA Ribossômico/metabolismo , Engenharia Genética , Humanos , Substâncias Macromoleculares , Metáfase , Proteínas Pol1 do Complexo de Iniciação de Transcrição/genética , Proteínas Pol1 do Complexo de Iniciação de Transcrição/fisiologia , Ligação Proteica , Transporte Proteico , Sequências Repetitivas de Ácido Nucleico , XenopusRESUMO
Isolated mesophyll cells from darkened leaves of the C(4) plant Digitaria sanguinalis keep functional plasmodesmata that allow the free exchange of low molecular mass compounds with the surrounding medium. This cell suspension system has been used to measure C(4) PEPC activity in situ using a spectrophotometric assay. Compared to the extracted enzyme assayed in vitro, the essentially non-phosphorylated 'in-cell' C(4) PEPC showed altered functional and regulatory properties. While the S (0.5) for PEP at pH 7.3 was only modestly changed (0.4-0.6 mM), the response to pH was shifted towards the acidic range, being close to the maximal value at pH 7.3. Using expected physiological concentrations of the metabolites, at pH 7.3, the IC(50) for malate showed a five-fold increase, from 1.5 to 8 mM, and was increased further to 22 mM in the presence of the allosteric activator glucose-6-phosphate (4 mM). Thiol compounds like DTT, mercaptoethanol and reduced glutathione weakened the in-situ sensitivity of C(4) PEPC to malate. However, none of them had any effect on this process in vitro. This was not due to thioredoxin-mediated or phoshorylation-dependent processes. Since glutathione is a physiological compound that is present mostly in the reduced state in the cell cytosol, a possible contribution of this thiol to the protection of the enzyme against malate in situ is proposed.
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Thus far, no transcription factor IIIA (TFIIIA) from higher plants has been cloned and characterized. We have cloned and characterized TFIIIA and ribosomal protein L5 from Arabidopsis thaliana. Primary sequence comparison revealed a high divergence of AtTFIIIA and a relatively high conservation of AtL5 when compared with other organisms. The AtTFIIIA cDNA encodes a protein with nine Cys(2)-His(2)-type zinc fingers, a 23 amino acid spacer between fingers 1 and 2, a 66 amino acid spacer between fingers 4 and 5, and a 50 amino acid non-finger C-terminal tail. Aside from the amino acids required for proper zinc finger folding, AtTFIIIA is highly divergent from other known TFIIIAs. AtTFIIIA can bind 5S rDNA, as well as 5S rRNA, and efficiently stimulates the transcription of an Arabidopsis 5S rRNA gene in vitro. AtL5 identity was confirmed by demonstrating that this protein binds to 5S rRNA but not to 5S rDNA. Protoplast transient expression assays with green fluorescent protein fusion proteins revealed that AtTFIIIA is absent from the cytoplasm and concentrated at several nuclear foci including the nucleolus. AtL5 protein accumulates in the nucleus, especially in the nucleolus, and is also present in the cytoplasm.