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Recent findings point to plant root traits as potentially important for shaping the boundaries of biomes and for maintaining the plant communities within. We examined two hypotheses: 1) Thin-rooted plant strategies might be favored in biomes with low soil resources; and 2) these strategies may act, along with fire, to maintain the sharp boundary between the Fynbos and Afrotemperate Forest biomes in South Africa. These biomes differ in biodiversity, plant traits, and physiognomy, yet exist as alternative stable states on the same geological substrate and in the same climate conditions. We conducted a 4-y field experiment to examine the ability of Forest species to invade the Fynbos as a function of growth-limiting nutrients and belowground plant-plant competition. Our results support both hypotheses: First, we found marked biome differences in root traits, with Fynbos species exhibiting the thinnest roots reported from any biome worldwide. Second, our field manipulation demonstrated that intense belowground competition inhibits the ability of Forest species to invade Fynbos. Nitrogen was unexpectedly the resource that determined competitive outcome, despite the long-standing expectation that Fynbos is severely phosphorus constrained. These findings identify a trait-by-resource feedback mechanism, in which most species possess adaptive traits that modify soil resources in favor of their own survival while deterring invading species. Our findings challenge the long-held notion that biome boundaries depend primarily on external abiotic constraints and, instead, identify an internal biotic mechanism-a selective feedback among traits, plant-plant competition, and ecosystem conditions-that, along with contrasting fire regime, can act to maintain biome boundaries.
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Ecossistema , Fenômenos Fisiológicos Vegetais , Raízes de Plantas/fisiologia , África do SulRESUMO
BACKGROUND: The field of genome editing has been revolutionized by the development of an easily programmable editing tool, the CRISPR-Cas9. Despite its promise, off-target activity of Cas9 posed a great disadvantage for genome editing purposes by causing DNA double strand breaks at off-target locations and causing unwanted editing outcomes. Furthermore, for gene integration applications, which introduce transgene sequences, integration of transgenes to off-target sites could be harmful, hard to detect, and reduce faithful genome editing efficiency. METHOD: Here we report the development of a multicolour fluorescence assay for studying CRISPR-Cas9-directed gene integration at an endogenous locus in human cell lines. We examine genetic integration of reporter genes in transiently transfected cells as well as puromycin-selected stable cell lines to determine the fidelity of multiple CRISPR-Cas9 strategies. RESULT: We found that there is a high occurrence of unwanted DNA integration which tarnished faithful knock-in efficiency. Integration outcomes are influenced by the type of DNA DSBs, donor design, the use of enhanced specificity Cas9 variants, with S-phase regulated Cas9 activity. Moreover, restricting Cas9 expression with a self-cleaving system greatly improves knock-in outcomes by substantially reducing the percentage of cells with unwanted DNA integration. CONCLUSION: Our results highlight the need for a more stringent assessment of CRISPR-Cas9-mediated knock-in outcomes, and the importance of careful strategy design to maximise efficient and faithful transgene integration.
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Sistemas CRISPR-Cas , Edição de Genes , Humanos , Edição de Genes/métodos , Quebras de DNA de Cadeia Dupla , Transgenes , DNARESUMO
PREMISE: Increases in genome size in plants-often associated with larger, low-density stomata and greater water-use efficiency (WUE)-could affect plant ecophysiological and hydraulic function. Variation in plant genome size is often due to polyploidy, having occurred repeatedly in the austral sedge genus Schoenus in the Cape Floristic Region (CFR), while species in the other major schoenoid genus in the region, Tetraria, have smaller genomes. Comparing these genera is useful as they co-occur at the landscape level, under broadly similar bioclimatic conditions. We hypothesized that CFR Schoenus have greater WUE, with lower maximum stomatal conductance (gwmax) imposed by larger, less-dense stomata. METHODS: We investigated relationships between genome size and stomatal parameters in a phylogenetic context, reconstructing a phylogeny of CFR-occurring Schoeneae (Cyperaceae). Species' stomatal and functional traits were measured from field-collected and herbarium specimens. Carbon stable isotopes were used as an index of WUE. Genome size was derived from flow-cytometric measurements of leafy shoots. RESULTS: Evolutionary regressions demonstrated that stomatal size and density covary with genome size, positively and negatively, respectively, with genome size explaining 72-75% of the variation in stomatal size. Larger-genomed species had lower gwmax and C:N ratios, particularly in culms. CONCLUSIONS: We interpret differences in vegetative physiology between the genera as evidence of more-conservative strategies in CFR Schoenus compared to the more-acquisitive Tetraria. Because Schoenus have smaller, reduced leaves, they likely rely more on culm photosynthesis than Tetraria. Across the CFR Schoeneae, ecophysiology correlates with genome size, but confounding sources of trait variation limit inferences about causal relationships between traits.
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Understanding climate change impacts on the Cape Floristic Region requires improved knowledge of plant physiological responses to the environment. Studies examining physiological responses of mountain fynbos have consisted of campaign-based measurements, capturing snapshots of plant water relations and photosynthesis. We examine conclusions drawn from prior studies by tracking in situ physiological responses of three species, representing dominant growth forms (proteoid, ericoid, restioid), over 2 years using miniature continuous sap flow technology, long-term observations of leaf/culm water potential and gas exchange, and xylem vulnerability to embolism. We observed considerable inter-specific variation in the timing and extent of seasonal declines in productivity. Shallow-rooted Erica monsoniana exhibited steep within-season declines in sap flow and water potentials, and pronounced inter-annual variability in total daily sap flux (Js). Protea repens showed steady reductions in Js across both years, despite deeper roots and less negative water potentials. Cannomois congesta-a shallow-rooted restioid-was least negatively impacted. Following rehydrating rain at the end of summer, gas exchange recovery was lower in the drier year compared with the normal year, but did not differ between species. Loss of function in the drier year was partially accounted for by loss of xylem transport capacity in Erica and Cannomois, but not Protea. Hitherto unseen water use patterns, including inter-annual variability of gas exchange associated with contrasting water uptake properties, reveal that species use different mechanisms to cope with summer dry periods. Revealing physiological responses of key growth forms enhances predictions of plant function within mountain fynbos under future conditions.
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Fenômenos Fisiológicos Vegetais , Água , Água/fisiologia , Folhas de Planta/fisiologia , Fotossíntese , Estações do Ano , Secas , Árvores/fisiologiaRESUMO
Growth plasticity may allow fire-prone species to maximize their recovery rates during temporary, sporadic periods of rainfall availability in the post-fire environment. However, moisture-driven growth plasticity could be maladaptive in nutrient-limited environments that require tighter control of growth and resource use. We investigated whether a trade-off between plasticity and conservatism mediates growth responses to altered rainfall seasonality in neighbouring shrubland communities that occupy different soils. We monitored post-fire vegetation regrowth in two structurally similar, Mediterranean-type shrublands for 3 years. We investigated the effects of experimentally altered rainfall seasonality on post-fire species' growth rates. We found that moisture-driven growth plasticity was higher among species occupying the fertile soils of the renosterveld site relative to those occupying the nutrient-poor soils of the fynbos site. This resulted in higher overall responsiveness of post-fire recovery patterns in renosterveld to experimental shifts in rainfall seasonality. In post-fire shrubland communities, the trade-off between moisture-dependent growth plasticity and resource conservatism could be mediated by soil nutrient availability. Therefore, edaphic differences between structurally similar shrublands could lead to differences in their sensitivity to post-fire rainfall seasonality.
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Incêndios , Ecossistema , SoloRESUMO
Most children with peripheral facial palsy will not have a cause identified. Although leukemia can cause facial nerve palsy, the magnitude of the risk is unknown and recommendations for investigations are variable. We are currently conducting a randomized, placebo-controlled trial of prednisolone for the treatment of Bell's palsy in children within the Paediatric Research in Emergency Departments International Collaborative emergency research network. In the course of the assessment for eligibility of the trial, from 644 acute-onset facial palsy presentations we identified 5 children with previously undiagnosed leukemia. We estimate the rate of leukemia in children with acute-onset facial palsy who present to emergency departments to be 0.6% (95% confidence interval 0.2% to 1.6%). In accordance with these cases, we suggest consideration of a screening CBC count for acute-onset peripheral facial palsy presentations in children before initiation of corticosteroid treatment.
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Paralisia de Bell/diagnóstico por imagem , Paralisia de Bell/etiologia , Glucocorticoides/uso terapêutico , Leucemia/complicações , Prednisolona/uso terapêutico , Austrália , Contagem de Células Sanguíneas , Criança , Pré-Escolar , Serviço Hospitalar de Emergência , Feminino , Humanos , Masculino , Nova Zelândia , RiscoRESUMO
BACKGROUND: Quality improvement systems are needed to overcome the 'Quality Gap' - difference between evidence-based guidelines and the care delivered. While there are a large array of potential quality assurance measures exists in the Paediatric Emergency Department, parent's/carer's perception of these is unknown. This study aimed to identify what 'quality of care' means to parents/carers of Paediatric Emergency Department (PED) patients, further determine which aspects of these are most important to them. Also, to identify which of the existing PED quality measures are most important to parents/carers, and their preferred method of providing feedback. METHODS: A Modified Rand-Delphi study was performed with parents/carers as the expert group and consensus was obtained from them via three web-based surveys. All parents/carers of children attending a tertiary paediatric hospital during six-week in winter were eligible- no exclusions. Quality measures scoring at least 7 on a 9-point Likert scale during the final survey were considered "very important", while those scoring at least an 8 were considered "extremely important". RESULTS: One hundred four parents/carers responded from a total of 1095 participants. Parents/carers generated 527 free text entries, to the initial survey on what 'quality of care' means. These were mapped to 48 quality measure which they ranked on subsequent surveys. Eighteen quality measures were considered very important by at least 90% of respondents. Of these, six were considered extremely important by at least 70% of respondents: 'Thorough medical assessment' (84%); 'A triage system' (84%); 'Experienced and knowledgeable staff that are skilled in paediatrics' (77%); 'Resources and equipment available to provide care' (72%); and 'Clear follow up plans and reviews that are communicated and scheduled' (72%). Parents/carers considered existing quality measures as important with 'timely treatment of a critical condition' as the most important. Most participants preferred to provide anonymous feedback (N = 69, 66%), online (N = 77, 72%) after discharge (N = 82, 70%). CONCLUSION: We have elicited what 'quality of care' means to parents/carers, and which aspects are most important to them. Parents/carers consider commonly used PED quality measure as very important. However, they are less important than outcomes generated by themselves. Further parents/carers in this study preferred to provide feedback that was anonymous and electronically distributed after they leave the ED.
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Cuidadores , Pais , Criança , Serviço Hospitalar de Emergência , Hospitais Pediátricos , Humanos , TriagemRESUMO
AIM: The number of children and young people presenting to emergency departments (EDs) with anxiety and depression is increasing. We aimed to determine parent perspectives on: (i) barriers to accessing non-ED mental health services; and (ii) improving access in the paediatric mental health service system. METHODS: Qualitative study with parents of children and young people aged 0-19 years who attended one of four EDs across Victoria between October 2017 and September 2018 and received a primary diagnosis of anxiety or depression. EXCLUSION CRITERIA: child or young person without a parent/guardian, or presented with self-harm or suicide attempt. Eligible participants completed semi-structured phone interviews. Interviews were recorded and transcripts were coded and analysed using content analysis. RESULTS: A total of 72 parents completed interviews. The average child age was 14 years (standard deviation 2.5) and two thirds identified as female (64%). A total of 57% of children and young people presented with a primary diagnosis of anxiety. Parents reported barriers in accessing care including: service shortages and inaccessibility, underresourced schools, lack of clinician mental health expertise, lack of child-clinician rapport, inconsistent care, financial constraints, lack of mental health awareness among parents, and stigma. Parents want expanded and improved access to services, more respite and support services, supportive schools, and improved mental health education for parents. CONCLUSIONS: Parents of children and young people attending the ED for anxiety and depression are generally dissatisfied with services for child mental health. Solutions that enable parents to better care for their child in the community are needed to improve care.
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Serviços de Saúde Mental , Adolescente , Adulto , Transtornos de Ansiedade , Criança , Pré-Escolar , Feminino , Acessibilidade aos Serviços de Saúde , Humanos , Lactente , Recém-Nascido , Saúde Mental , Pais , Pesquisa Qualitativa , Adulto JovemRESUMO
OBJECTIVES: Stable isotope analysis has been used to investigate consumption of marine resources in a variety of terrestrial mammals, including humans, but not yet in extant nonhuman primates. We sought to test the efficacy of stable isotope analysis as a tool for such studies by comparing isotope- and observation-based estimates of marine food consumption by a troop of noncommensal, free-ranging chacma baboons. MATERIALS AND METHODS: We determined δ13 C and δ15 N values of baboon hair (n = 9) and fecal samples (n = 144), and principal food items (n = 362). These values were used as input for diet models, the outputs of which were compared to observation-based estimates of marine food consumption. RESULTS: Fecal δ13 C values ranged from -29.3 to -25.6. δ15 N values ranged from 0.9 to 6.3 and were positively correlated with a measure of marine foraging during the dietary integration period. Mean (± SD) δ13 C values of adult male and female baboon hairs were -21.6 (± 0.1) and -21.8 (± 0.3) respectively, and corresponding δ15 N values were 5.0 (± 0.3) and 3.9 (± 0.2). Models indicated that marine contributions were ≤10% of baboon diet within any season, and contributed ≤17% of dietary protein through the year. DISCUSSION: Model output and observational data were in agreement, both indicating that despite their abundance in the intertidal region, marine foods comprised only a small proportion of baboon diet. This suggests that stable isotope analysis is a viable tool for investigating marine food consumption by natural-foraging primates in temperate regions.
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Isótopos de Carbono/análise , Dieta/estatística & dados numéricos , Dieta/veterinária , Comportamento Alimentar/fisiologia , Isótopos de Nitrogênio/análise , Papio ursinus/fisiologia , Animais , Antropologia Física , Fezes/química , Feminino , Alimentos/estatística & dados numéricos , Cabelo/química , Masculino , África do SulRESUMO
Attempts to understand mechanisms underlying plant mortality during drought have led to the emergence of a hydraulic framework describing distinct hydraulic strategies among coexisting species. This framework distinguishes species that rapidly decrease stomatal conductance (gs), thereby maintaining high water potential (Px; isohydric), from those species that maintain relatively high gs at low Px, thereby maintaining carbon assimilation, albeit at the cost of loss of hydraulic conductivity (anisohydric). This framework is yet to be tested in biodiverse communities, potentially due to a lack of standardized reference values upon which hydraulic strategies can be defined. We developed a system of quantifying hydraulic strategy using indices from vulnerability curves and stomatal dehydration response curves and tested it in a speciose community from South Africa's Cape Floristic Region. Degree of stomatal regulation over cavitation was defined as the margin between Px at stomatal closure (Pg12) and Px at 50% loss of conductivity. To assess relationships between hydraulic strategy and mortality mechanisms, we developed proxies for carbon limitation and hydraulic failure using time since Pg12 and loss of conductivity at minimum seasonal Px, respectively. Our approach captured continuous variation along an isohydry/anisohydry axis and showed that this variation was linearly related to xylem safety margin. Degree of isohydry/anisohydry was associated with contrasting predictions for mortality during drought. Merging stomatal regulation strategies that represent an index of water use behavior with xylem vulnerability facilitates a more comprehensive framework with which to characterize plant response to drought, thus opening up an avenue for predicting the response of diverse communities to future droughts.
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Secas , Estômatos de Plantas/fisiologia , Xilema/fisiologia , Biodiversidade , Carbono/química , Dessecação , Folhas de Planta/fisiologia , Fenômenos Fisiológicos Vegetais , Brotos de Planta/fisiologia , Caules de Planta/fisiologia , Transpiração Vegetal/fisiologia , Pressão , Estações do Ano , África do Sul , ÁguaRESUMO
Increases in woody plant cover in savanna grassland environments have been reported on globally for over 50 years and are generally perceived as a threat to rangeland productivity and biodiversity. Despite this, few attempts have been made to estimate the extent of woodland increase at a national scale, principally due to technical constraints such as availability of appropriate remote sensing products. In this study, we aimed to measure the extent to which woodlands have replaced grasslands in South Africa's grassy biomes. We use multiseason Landsat data in conjunction with satellite L-band radar backscatter data to estimate the extent of woodlands and grasslands in 1990 and 2013. The method employed allows for a unique, nationwide measurement of transitions between grassland and woodland classes in recent decades. We estimate that during the 23-year study period, woodlands have replaced grasslands over ~57 000 km2 and conversely that grasslands have replaced woodlands over ~30 000 km2 , a net increase in the extent of woodland of ~27 000 km2 and an annual increase of 0.22%. The changes varied markedly across the country; areas receiving over 500 mm mean annual precipitation showed higher rates of woodland expansion than regions receiving <500 mm (0.31% yr-1 and 0.11% yr-1 , respectively). Protected areas with elephants showed clear loss of woodlands (-0.43% yr-1 ), while commercial rangelands and traditional rangelands showed increases in woodland extent (>0.19% yr-1 ). The woodland change map presented here provides a unique opportunity to test the numerous models of woody plant encroachment at a national/regional scale.
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Conservação dos Recursos Naturais , Florestas , Poaceae , Ecossistema , Plantas , Dinâmica Populacional , África do Sul , ÁrvoresRESUMO
AIM: To compare the parental motivators and referring general practitioner's (GP's) reasons for advising emergency department (ED) attendance with the assessment of ED medical staff. To compare ED clinician opinion with other published methods that have attempted to define 'primary care suitable' presentations to the ED. METHODS: A prospective observational study and series of surveys regarding the attendance of children presenting to a single tertiary paediatric ED. Surveys were distributed to the treating ED clinician, the child's parent/guardian, and the referring GP. Results between the three groups were analysed and compared. RESULTS: There were a total of 1069 presentations during the study period. Six hundred (58.4%, 95% CI 55.3-61.4%) presentations were judged as 'ED appropriate' by the treating ED clinician. When compared with methods used to retrospectively judge whether ED patients are considered 'primary care suitable', ED clinicians disagree between 22.4 and 38.8% of the time. For patients who presented directly to ED, 85.6% did so for a medical reason, whilst 32.1% did so for a GP access reason. Being referred by a GP improved the ED clinicians' opinion of the appropriateness of the presentation (49.2 vs. 73.9%, P < 0.05). CONCLUSIONS: We caution that many strategies attempting to 'solve' the issue of increasing ED attendances by paediatric patients have been driven by opinion, and a better understanding of the motivators that drive this behaviour is needed. We believe the solution to increasing utilisation of EDs by children must be a balanced approach that addresses community expectations and appropriately resources EDs.
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Atitude Frente a Saúde , Serviço Hospitalar de Emergência/estatística & dados numéricos , Motivação , Pais/psicologia , Atenção Primária à Saúde/estatística & dados numéricos , Encaminhamento e Consulta/estatística & dados numéricos , Adolescente , Criança , Pré-Escolar , Feminino , Medicina Geral , Pesquisas sobre Atenção à Saúde , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Humanos , Lactente , Recém-Nascido , Masculino , Estudos Prospectivos , VitóriaRESUMO
Recent work suggests that hydraulic mechanisms, rather than cambium necrosis, may account for rapid post-fire tree mortality. We experimentally tested for xylem cavitation, as a result of exposure to high-vapour-deficit (D) heat plumes, and permanent xylem deformation, as a result of thermal softening of lignin, in two tree species differing in fire tolerance. We measured percentage loss of conductance (PLC) in distal branches that had been exposed to high-D heat plumes or immersed in hot water baths (high temperature, but not D). Results were compared with predictions from a parameterized hydraulic model. Physical damage to the xylem was examined microscopically. Both species suffered c. 80% PLC when exposed to a 100°C plume. However, at 70°C, the fire-sensitive Kiggelaria africana suffered lower PLC (49%) than the fire-resistant Eucalytpus cladocalyx (80%). Model simulations suggested that differences in PLC between species were a result of greater hydraulic segmentation in E. cladocalyx. Kiggelaria africana suffered considerable PLC (59%), as a result of heat-induced xylem deformation, in the water bath treatments, but E. cladocalyx did not. We suggest that a suite of 'pyrohydraulic' traits, including hydraulic segmentation and heat sensitivity of the xylem, may help to explain why some tree species experience rapid post-fire mortality after low-intensity fires and others do not.
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Temperatura Alta , Árvores/fisiologia , Xilema/fisiologia , Análise de Variância , Simulação por Computador , Eucalyptus/fisiologia , Modelos Biológicos , Folhas de Planta/fisiologia , Fatores de Tempo , Xilema/ultraestruturaRESUMO
The famous nonmetal-to-metal transition in Na-ammonia solutions is investigated in nanoscale solution droplets by photoelectron spectroscopy. In agreement with the bulk solutions, a strong indication for a transition to the metallic state is found at an average metal concentration of 8.8±2.2â mole%. The smallest entity for the phase transition to be observed consists of approximately 100-200 solvent molecules. The quantification of this critical entity size is a stepping stone toward a deeper understanding of these quantum-classical solutions through direct modeling at the molecular level.
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Many species have the ability to resprout vegetatively after a substantial loss of biomass induced by environmental stress, including drought. Many of the regions characterised by ecosystems where resprouting is common are projected to experience more frequent and intense drought during the 21st Century. However, in assessments of ecosystem response to drought disturbance there has been scant consideration of the resilience and post-drought recovery of resprouting species. Systematic differences in hydraulic and allocation traits suggest that resprouting species are more resilient to drought-stress than nonresprouting species. Evidence suggests that ecosystems dominated by resprouters recover from disturbance more quickly than ecosystems dominated by nonresprouters. The ability of resprouters to avoid mortality and withstand drought, coupled with their ability to recover rapidly, suggests that the impact of increased drought stress in ecosystems dominated by these species may be small. The strategy of resprouting needs to be modelled explicitly to improve estimates of future climate-change impacts on the carbon cycle, but this will require several important knowledge gaps to be filled before resprouting can be properly implemented.
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Secas , Desenvolvimento Vegetal , Fenômenos Fisiológicos Vegetais , Regeneração/fisiologia , Ecossistema , Estresse FisiológicoRESUMO
TAL1 is an important regulator of hematopoiesis and its expression is tightly controlled despite complexities in its genomic organization. It is frequently misregulated in T-cell acute lymphoblastic leukemia (T-ALL), often due to deletions between TAL1 and the neighboring STIL gene. To better understand the events that lead to TAL1 expression in hematopoiesis and in T-ALL, we studied looping interactions at the TAL1 locus. In TAL1-expressing erythroid cells, the locus adopts a looping "hub" which brings into close physical proximity all known TAL1 cis-regulatory elements including CTCF-bound insulators. Loss of GATA1 results in disassembly of the hub and loss of CTCF/RAD21 from one of its insulators. Genes flanking TAL1 are partly dependent on hub integrity for their transcriptional regulation. We identified looping patterns unique to TAL1-expressing T-ALL cells, and, intriguingly, loops occurring between the TAL1 and STIL genes at the common TAL1/STIL breakpoints found in T-ALL. These findings redefine how TAL1 and neighboring genes communicate within the nucleus, and indicate that looping facilitates both normal and aberrant TAL1 expression and may predispose to structural rearrangements in T-ALL. We also propose that GATA1-dependent looping mechanisms may facilitate the conservation of TAL1 regulation despite cis-regulatory remodeling during vertebrate evolution.
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Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Cromatina/genética , Regulação Neoplásica da Expressão Gênica , Leucemia-Linfoma Linfoblástico de Células T Precursoras/genética , Proteínas Proto-Oncogênicas/genética , Animais , Cromatina/química , Elementos Facilitadores Genéticos/genética , Fator de Transcrição GATA1/genética , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Células Jurkat , Células K562 , Linfócitos/citologia , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos ICR , Núcleosídeo-Fosfato Quinase/genética , Regiões Promotoras Genéticas/genética , Conformação Proteica , Proteína 1 de Leucemia Linfocítica Aguda de Células TRESUMO
The nuclear genomes of vertebrates show a highly organized program of DNA replication where GC-rich isochores are replicated early in S-phase, while AT-rich isochores are late replicating. GC-rich regions are gene dense and are enriched for active transcription, suggesting a connection between gene regulation and replication timing. Insulator elements can organize independent domains of gene transcription and are suitable candidates for being key regulators of replication timing. We have tested the impact of inserting a strong replication origin flanked by the ß-globin HS4 insulator on the replication timing of naturally late replicating regions in two different avian cell types, DT40 (lymphoid) and 6C2 (erythroid). We find that the HS4 insulator has the capacity to impose a shift to earlier replication. This shift requires the presence of HS4 on both sides of the replication origin and results in an advance of replication timing of the target locus from the second half of S-phase to the first half when a transcribed gene is positioned nearby. Moreover, we find that the USF transcription factor binding site is the key cis-element inside the HS4 insulator that controls replication timing. Taken together, our data identify a combination of cis-elements that might constitute the basic unit of multi-replicon megabase-sized early domains of DNA replication.
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Replicação do DNA , Elementos Isolantes , Origem de Replicação , Fatores Estimuladores Upstream/metabolismo , Acetilação , Alelos , Animais , Sítios de Ligação , Linhagem Celular , Galinhas/genética , Galinhas/metabolismo , Imunoprecipitação da Cromatina , Mapeamento Cromossômico , DNA/genética , DNA/metabolismo , Células Eritroides/citologia , Células Eritroides/metabolismo , Histonas/genética , Histonas/metabolismo , Linfócitos/citologia , Linfócitos/metabolismo , Mutagênese Insercional , Fase S , Fatores de Tempo , Transcrição Gênica , Ativação Transcricional , Transfecção , Transgenes , Fatores Estimuladores Upstream/genética , Globinas beta/genética , Globinas beta/metabolismoRESUMO
This paper reports on a curiosity concerning magic numbers in neutral molecular clusters, namely on magic numbers related to the photoelectron anisotropy in angle-resolved photoelectron spectra. With a combination of density functional calculations and experiment, we search for magic numbers in Na(H2O)n, Na(NH3)n, Na(CH3OH)n, and Na(CH3OCH3)n clusters. In clusters of high symmetry, the highest occupied molecular orbital can be delocalized over an extended region, forming a symmetric charge distribution of high s character, which results in a pronounced anisotropy in the photoelectron angular distribution. We find magic numbers at n = 6 and 4 for sodium-doped dimethyl ether and ammonia clusters, respectively, but not for sodium-doped water and methanol clusters, which is likely a consequence of the degree of hydrogen bonding and the number of structural isomers.
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We present a combined computational and threshold photoelectron photoion coincidence study of two isotopologues of dimethyl ether, (DME - h6)n and (DME - d6)n n = 1 and 2, in the 9-14 eV photon energy range. Multiple isomers of neutral dimethyl ether dimer were considered, all of which may be present, and exhibited varying C-Hâ¯O interactions. Results from electronic structure calculations predict that all of them undergo barrierless proton transfer upon photoionization to the ground electronic state of the cation. In fact, all neutral isomers were found to relax to the same radical cation structure. The lowest energy dissociative photoionization channel of the dimer leads to CH3OHCH3 (+) by the loss of CH2OCH3 with a 0 K appearance energy of 9.71 ± 0.03 eV and 9.73 ± 0.03 eV for (DME - h6)2 and deuterated (DME - d6)2, respectively. The ground state threshold photoelectron spectrum band of the dimethyl ether dimer is broad and exhibits no vibrational structure. Dimerization results in a 350 meV decrease of the valence band appearance energy, a 140 meV decrease of the band maximum, thus an almost twofold increase in the ground state band width, compared with DME - d6 monomer.
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Ligação de Hidrogênio , Éteres Metílicos/química , Prótons , Carbono/química , Cátions/química , Simulação por Computador , Dimerização , Hidrogênio/química , Isomerismo , Modelos Químicos , Oxigênio/química , Processos Fotoquímicos , Espectroscopia Fotoeletrônica , Fótons , TemperaturaRESUMO
We propose angle-resolved photoelectron spectroscopy of aerosol particles as an alternative way to determine the electron mean free path of low energy electrons in solid and liquid materials. The mean free path is obtained from fits of simulated photoemission images to experimental ones over a broad range of different aerosol particle sizes. The principal advantage of the aerosol approach is twofold. First, aerosol photoemission studies can be performed for many different materials, including liquids. Second, the size-dependent anisotropy of the photoelectrons can be exploited in addition to size-dependent changes in their kinetic energy. These finite size effects depend in different ways on the mean free path and thus provide more information on the mean free path than corresponding liquid jet, thin film, or bulk data. The present contribution is a proof of principle employing a simple model for the photoemission of electrons and preliminary experimental data for potassium chloride aerosol particles.