975 research outputs found
A microfluidic processor for gene expression profiling of single human embryonic stem cells
The gene expression of human embryonic stem cells (hESC) is a critical aspect for understanding the normal and pathological development of human cells and tissues. Current bulk gene expression assays rely on RNA extracted from cell and tissue samples with various degree of cellular heterogeneity. These cell population averaging data are difficult to interpret, especially for the purpose of understanding the regulatory relationship of genes in the earliest phases of development and differentiation of individual cells. Here, we report a microfluidic approach that can extract total mRNA from individual single-cells and synthesize cDNA on the same device with high mRNA-to-cDNA efficiency. This feature makes large-scale single-cell gene expression profiling possible. Using this microfluidic device, we measured the absolute numbers of mRNA molecules of three genes (B2M, Nodal and Fzd4) in a single hESC. Our results indicate that gene expression data measured from cDNA of a cell population is not a good representation of the expression levels in individual single cells. Within the G0/G1 phase pluripotent hESC population, some individual cells did not express all of the 3 interrogated genes in detectable levels. Consequently, the relative expression levels, which are broadly used in gene expression studies, are very different between measurements from population cDNA and single-cell cDNA. The results underscore the importance of discrete single-cell analysis, and the advantages of a microfluidic approach in stem cell gene expression studies
A Simple Procedure for Constructing 5'-Amino-Terminated Oligodeoxynucleotides in Aqueous Solution
A rapid method for the synthesis of oligodeoxynucleotides (ODNs) terminated by 5'-amino-5'-deoxythymidine is described. A 3'-phosphorylated ODN (the donor) is incubated in aqueous solution with 5'-amino- 5'-deoxythymidine in the presence of N-(3-dimethylaminopropyl)-)N'-ethylcarbodiimide hydrochloride (EDC), extending the donor by one residue via a phosphoramidate bond. Template- directed ligation of the extended donor and an acceptor ODN, followed by acid hydrolysis, yields the acceptor ODN extended by a single 5'-amino-5'-deoxythymidine residue at its 5'terminus
Phytosociological analysis of restored and managed grassland habitat within an urban national park
Floyd Bennett Field (FBF), 579 ha in extent, is a division of Gateway National Recreation Area. It is the site of a former airfield, constructed by filling salt marshes with dredged materials. Except for the portion known locally as the “North Forty,” all sections of FBF have been cut over to maintain low vegetation. A grassland management plan (GRAMP) for 165 ha was initiated in 1986, to maintain habitats for open-country birds. Over the next few years, encroaching woody vegetation was removed manually and mechanically from the management area. Since then, it has been maintained as a grassland and receives annual mowing, as well as continued manual removal of the larger woody sprouts.
A portion of the GRAMP management area (III) was selected for intensive study of vegetation composition. A grid system was created and vegetation cover was estimated in 127, 1 m × 1 m quadrats. The quadrats were subjected to cluster analysis (CA). Eleven clusters were recognized. These clusters were treated as “plant associations.” The following types were distinguished: (native) little bluestem–dewberry grassland, six-weeks fescue annual grassland, a grass marsh, a rush marsh, a switchgrass dry grassland, and a deer-tongue panicgrass grassland; (exotic) mugwort herbland, oriental bittersweet-Japanese honeysuckle vineland, Kentucky bluegrass-mixed grassland, Japanese knotweed tall herbland, and spotted knapweed-common St. Johnswort herbland. The little bluestem–dewberry association accounted for nearly half of all quadrats; six subclusters were recognized. The plant associations determined by CA were compared with plant lists compiled during traverses of all of the map categories in the six GRAMP Areas (I, II, III, IV, V, VI). A table was created to relate the quantitative data of the plant associations to the appropriate map categories. A nonmetric multidimensional scaling ordination (NMDS) was performed on the quadrat data. Finally, the plant associations were compared with those described in the literature of local vegetation studies. The mowing program has been effective in decreasing woody plant cover and has permitted the invasion of a few taxa into monospecific communities, but attendant disturbance of the substrate is likely to cause an increase in exotic plant taxa. As earlier studies noted, mowing has caused the increase in cover of sod-forming grass, and bare ground has virtually disappeared in the managed area. This has negative implications for the maintenance of those grassland bird species that require open ground for nesting
Two Earth-sized planets orbiting Kepler-20
Since the discovery of the first extrasolar giant planets around Sun-like
stars, evolving observational capabilities have brought us closer to the
detection of true Earth analogues. The size of an exoplanet can be determined
when it periodically passes in front of (transits) its parent star, causing a
decrease in starlight proportional to its radius. The smallest exoplanet
hitherto discovered has a radius 1.42 times that of the Earth's radius (R
Earth), and hence has 2.9 times its volume. Here we report the discovery of two
planets, one Earth-sized (1.03R Earth) and the other smaller than the Earth
(0.87R Earth), orbiting the star Kepler-20, which is already known to host
three other, larger, transiting planets. The gravitational pull of the new
planets on the parent star is too small to measure with current
instrumentation. We apply a statistical method to show that the likelihood of
the planetary interpretation of the transit signals is more than three orders
of magnitude larger than that of the alternative hypothesis that the signals
result from an eclipsing binary star. Theoretical considerations imply that
these planets are rocky, with a composition of iron and silicate. The outer
planet could have developed a thick water vapour atmosphere.Comment: Letter to Nature; Received 8 November; accepted 13 December 2011;
Published online 20 December 201
Nanometer Scale Dielectric Fluctuations at the Glass Transition
Using non-contact scanning probe microscopy (SPM) techniques, dielectric
properties were studied on 50 nanometer length scales in poly-vinyl-acetate
(PVAc) films in the vicinity of the glass transition. Low frequency (1/f) noise
observed in the measurements, was shown to arise from thermal fluctuations of
the electric polarization. Anomalous variations observed in the noise spectrum
provide direct evidence for cooperative nano-regions with heterogeneous
kinetics. The cooperative length scale was determined. Heterogeneity was
long-lived only well below the glass transition for faster than average
processes.Comment: 4 pages, 4 embedded PS figures, RevTeX - To appear in Phys. Rev. Let
Kepler Flares II: The Temporal Morphology of White-Light Flares on GJ 1243
We present the largest sample of flares ever compiled for a single M dwarf,
the active M4 star GJ 1243. Over 6100 individual flare events, with energies
ranging from to erg, are found in 11 months of 1-minute
cadence data from Kepler. This sample is unique for its completeness and
dynamic range. We have developed automated tools for finding flares in
short-cadence Kepler light curves, and performed extensive validation and
classification of the sample by eye. From this pristine sample of flares we
generate a median flare template. This template shows that two exponential
cooling phases are present during the white-light flare decay, providing
fundamental constraints for models of flare physics. The template is also used
as a basis function to decompose complex multi-peaked flares, allowing us to
study the energy distribution of these events. Only a small number of flare
events are not well fit by our template. We find that complex, multi-peaked
flares occur in over 80% of flares with a duration of 50 minutes or greater.
The underlying distribution of flare durations for events 10 minutes and longer
appears to follow a broken power law. Our results support the idea that
sympathetic flaring may be responsible for some complex flare events.Comment: 12 pages, 9 figures, accepted for publication in Ap
Study protocol for a type III hybrid effectiveness-implementation trial of strategies to implement firearm safety promotion as a universal suicide prevention strategy in pediatric primary care
BACKGROUND: Insights from behavioral economics, or how individuals\u27 decisions and behaviors are shaped by finite cognitive resources (e.g., time, attention) and mental heuristics, have been underutilized in efforts to increase the use of evidence-based practices in implementation science. Using the example of firearm safety promotion in pediatric primary care, which addresses an evidence-to-practice gap in universal suicide prevention, we aim to determine: is a less costly and more scalable behavioral economic-informed implementation strategy (i.e., Nudge ) powerful enough to change clinician behavior or is a more intensive and expensive facilitation strategy needed to overcome implementation barriers?
METHODS: The Adolescent and child Suicide Prevention in Routine clinical Encounters (ASPIRE) hybrid type III effectiveness-implementation trial uses a longitudinal cluster randomized design. We will test the comparative effectiveness of two implementation strategies to support clinicians\u27 use of an evidence-based firearm safety practice, S.A.F.E. Firearm, in 32 pediatric practices across two health systems. All pediatric practices in the two health systems will receive S.A.F.E. Firearm materials, including training and cable locks. Half of the practices (k = 16) will be randomized to receive Nudge; the other half (k = 16) will be randomized to receive Nudge plus 1 year of facilitation to target additional practice and clinician implementation barriers (Nudge+). The primary implementation outcome is parent-reported clinician fidelity to the S.A.F.E Firearm program. Secondary implementation outcomes include reach and cost. To understand how the implementation strategies work, the primary mechanism to be tested is practice adaptive reserve, a self-report practice-level measure that includes relationship infrastructure, facilitative leadership, sense-making, teamwork, work environment, and culture of learning.
DISCUSSION: The ASPIRE trial will integrate implementation science and behavioral economic approaches to advance our understanding of methods for implementing evidence-based firearm safety promotion practices in pediatric primary care. The study answers a question at the heart of many practice change efforts: which strategies are sufficient to support change, and why? Results of the trial will offer valuable insights into how best to implement evidence-based practices that address sensitive health matters in pediatric primary care.
TRIAL REGISTRATION: ClinicalTrials.gov, NCT04844021 . Registered 14 April 2021
3DKL v1.0: creating the first 3D geological model of Kuala Lumpur
The objective of UN Sustainable Development Goal 11 is to make cities and human settlements inclusive, safe, resilient and sustainable. Geoscience can play a significant role in achieving targets within this goal by developing a better understanding of geological properties and processes within urban environments, and by ensuring that this understanding is integrated into urban development. A key step in this process will be enhancing awareness of urban geology among non-geoscience decision-makers, so that inherent subsurface risks and benefits are understood and accounted for during all phases of development. Three-dimensional geological models are an effective tool for geologists to communicate with stakeholders in government and industry during that process. They can also provide a framework to enable geological data and information to be integrated into Building and City Information Models, and thus facilitate more effective infrastructure and utility asset management. This paper describes the modelling workflow adopted by a consortium of geoscientists from government, industry and academia to deliver the first 3D geological model of Kuala Lumpur – 3DKL v1.0. The modelling workflow involved: digitising borehole logs from site investigation reports and storing them in a dedicated geospatially-enabled SQLite borehole database; viewing and interpreting that borehole data using QGIS software; generating multiple orthogonally oriented cross-section profiles across the modelled area using Groundhog Desktop software; and integrating the information derived from the interpreted boreholes, surface data and cross-section profiles to generate a 3D geological model in Leapfrog Geo software. 3DKL v1.0 has demonstrated proof-of-concept: we have developed a workflow, based largely on freely-available software, for transforming borehole information, previously captured in paper records, into a conceptual 3D model. The modelling process has also identified areas where geological knowledge and data need to be enhanced if 3DKL is to fulfil its potential to support more sustainable and resilient urban development in Kuala Lumpur
Phase 2 Study of the Safety and Tolerability of Maraviroc-Containing Regimens to Prevent HIV Infection in Men Who Have Sex With Men (HPTN 069/ACTG A5305)
Maraviroc (MVC) is a candidate for human immunodeficiency virus (HIV) pre-exposure prophylaxis
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