357 research outputs found

    Implication of polymerase recycling for nascent transcript quantification by live cell imaging.

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    Transcription enables the production of RNA from a DNA template. Due to the highly dynamic nature of transcription, live-cell imaging methods play a crucial role in measuring the kinetics of this process. For instance, transcriptional bursts have been visualized using fluorescent phage-coat proteins that associate tightly with messenger RNA (mRNA) stem loops formed on nascent transcripts. To convert the signal emanating from a transcription site into meaningful estimates of transcription dynamics, the influence of various parameters on the measured signal must be evaluated. Here, the effect of gene length on the intensity of the transcription site focus was analyzed. Intuitively, a longer gene can support a larger number of transcribing polymerases, thus leading to an increase in the measured signal. However, measurements of transcription induced by hyper-osmotic stress responsive promoters display independence from gene length. A mathematical model of the stress-induced transcription process suggests that the formation of gene loops that favor the recycling of polymerase from the terminator to the promoter can explain the observed behavior. One experimentally validated prediction from this model is that the amount of mRNA produced from a short gene should be higher than for a long one as the density of active polymerase on the short gene will be increased by polymerase recycling. Our data suggest that this recycling contributes significantly to the expression output from a gene and that polymerase recycling is modulated by the promoter identity and the cellular state

    Wakeful rest alleviates interference-based forgetting

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    Retroactive interference (RI)—the disruptive influence of events occurring after the formation of a new memory—is one of the primary causes of forgetting. Placing individuals within an environment that postpones interference should, therefore, greatly reduce the likelihood of information being lost from memory. For example, a short period of wakeful rest should diminish interference-based forgetting. To test this hypothesis, participants took part in a foreign language learning activity and were shown English translations of 20 Icelandic words for immediate recall. Half of the participants were then given an 8-min rest before completing a similar or dissimilar interfering distractor task. The other half did not receive a rest until after the distractor task, at which point interference had already taken place. All participants were then asked to translate the Icelandic words for a second time. Results revealed that retention was significantly worse at the second recall test, but being allowed a brief rest before completing the distractor task helped reduce the amount of forgetting. Taking a short, passive break can shield new memories from RI and alleviate forgetting.ERAS Scheme, University of Wolverhampto

    Maximal entropy inference of oncogenicity from phosphorylation signaling

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    Point mutations in the phosphorylation domain of the Bcr-Abl fusion oncogene give rise to drug resistance in chronic myelogenous leukemia patients. These mutations alter kinase-mediated signaling function and phenotypic outcome. An information theoretic analysis of the correlation of phosphoproteomic profiling and transformation potency of the oncogene in different mutants is presented. The theory seeks to predict the leukemic transformation potency from the observed signaling by constructing a distribution of maximal entropy of site-specific phosphorylation events. The theory is developed with special reference to systems biology where high throughput measurements are typical. We seek sets of phosphorylation events most contributory to predicting the phenotype by determining the constraints on the signaling system. The relevance of a constraint is measured by how much it reduces the value of the entropy from its global maximum, where all events are equally likely. Application to experimental phospho-proteomics data for kinase inhibitor-resistant mutants shows that there is one dominant constraint and that other constraints are not relevant to a similar extent. This single constraint accounts for much of the correlation of phosphorylation events with the oncogenic potency and thereby usefully predicts the trends in the phenotypic output. An additional constraint possibly accounts for biological fine structure

    Changes in lower limb rotation after soft tissue surgery in spastic diplegia: 3-dimensional gait analysis in 28 children

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    Background and purpose Rotational osteotomies are usually necessary to correct pronounced rotational deformities in ambulant children with cerebral palsy. The effects of soft tissue surgery on such deformities are unclear. In this retrospective study, we determined whether multilevel soft tissue surgery, performed to correct deformities in the sagittal plane, would also have an effect on rotational parameters

    Incorporating field wind data to improve crop evapotranspiration parameterization in heterogeneous regions

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    Accurate parameterization of reference evapotranspiration ( ET0) is necessary for optimizing irrigation scheduling and avoiding costs associated with over-irrigation (water expense, loss of water productivity, energy costs, and pollution) or with under-irrigation (crop stress and suboptimal yields or quality). ET0 is often estimated using the FAO-56 method with meteorological data gathered over a reference surface, usually short grass. However, the density of suitable ET0 stations is often low relative to the microclimatic variability of many arid and semi-arid regions, leading to a potentially inaccurate ET0 for irrigation scheduling. In this study, we investigated multiple ET0 products from six meteorological stations, a satellite ET0 product, and integration (merger) of two stations’ data in Southern California, USA. We evaluated ET0 against lysimetric ET observations from two lysimeter systems (weighing and volumetric) and two crops (wine grapes and Jerusalem artichoke) by calculating crop ET ( ETc) using crop coefficients for the lysimetric crops with the different ET0. ETc calculated with ET0 products that incorporated field-specific wind speed had closer agreement with lysimetric ET, with RMSE reduced by 36 and 45% for grape and Jerusalem artichoke, respectively, with on-field anemometer data compared to wind data from the nearest station. The results indicate the potential importance of on-site meteorological sensors for ET0 parameterization; particularly where microclimates are highly variable and/or irrigation water is expensive or scarce

    Reviews and syntheses : Turning the challenges of partitioning ecosystem evaporation and transpiration into opportunities

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    Evaporation (E) and transpiration (T) respond differently to ongoing changes in climate, atmospheric composition, and land use. It is difficult to partition ecosystem-scale evapotranspiration (ET) measurements into E and T, which makes it difficult to validate satellite data and land surface models. Here, we review current progress in partitioning E and T and provide a prospectus for how to improve theory and observations going forward. Recent advancements in analytical techniques create new opportunities for partitioning E and T at the ecosystem scale, but their assumptions have yet to be fully tested. For example, many approaches to partition E and T rely on the notion that plant canopy conductance and ecosystem water use efficiency exhibit optimal responses to atmospheric vapor pressure deficit (D). We use observations from 240 eddy covariance flux towers to demonstrate that optimal ecosystem response to D is a reasonable assumption, in agreement with recent studies, but more analysis is necessary to determine the conditions for which this assumption holds. Another critical assumption for many partitioning approaches is that ET can be approximated as T during ideal transpiring conditions, which has been challenged by observational studies. We demonstrate that T can exceed 95% of ET from certain ecosystems, but other ecosystems do not appear to reach this value, which suggests that this assumption is ecosystem-dependent with implications for partitioning. It is important to further improve approaches for partitioning E and T, yet few multi-method comparisons have been undertaken to date. Advances in our understanding of carbon-water coupling at the stomatal, leaf, and canopy level open new perspectives on how to quantify T via its strong coupling with photosynthesis. Photosynthesis can be constrained at the ecosystem and global scales with emerging data sources including solar-induced fluorescence, carbonyl sulfide flux measurements, thermography, and more. Such comparisons would improve our mechanistic understanding of ecosystem water fluxes and provide the observations necessary to validate remote sensing algorithms and land surface models to understand the changing global water cycle.Peer reviewe
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