256 research outputs found

    Quantification of total T-cell receptor diversity by flow cytometry and spectratyping

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    BACKGROUND: T-cell receptor diversity correlates with immune competency and is of particular interest in patients undergoing immune reconstitution. Spectratyping generates data about T-cell receptor CDR3 length distribution for each BV gene but is technically complex. Flow cytometry can also be used to generate data about T-cell receptor BV gene usage, but its utility has not been compared to or tested in combination with spectratyping. RESULTS: Using flow cytometry and spectratype data, we have defined a divergence metric that quantifies the deviation from normal of T-cell receptor repertoire. We have shown that the sample size is a sensitive parameter in the predicted flow divergence values, but not in the spectratype divergence values. We have derived two ways to correct for the measurement bias using mathematical and statistical approaches and have predicted a lower bound in the number of lymphocytes needed when using the divergence as a substitute for diversity. CONCLUSIONS: Using both flow cytometry and spectratyping of T-cells, we have defined the divergence measure as an indirect measure of T-cell receptor diversity. We have shown the dependence of the divergence measure on the sample size before it can be used to make predictions regarding the diversity of the T-cell receptor repertoire

    Inverted Fluorescence Microscope

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    Team F13 is composed of Trevor Blythe, Spencer Hann, Matthew Pfeiffer, and Thomas Eggenberger. We are all majoring in mechanical engineering and in our final year of study here at Cal Poly San Luis Obispo. This project is a continuation of a 2019-2020 senior project. The previous team designed and built a functioning inverted fluorescence microscope (IFM) from scratch. This device was created as a lab tool for undergraduate students to be able to perform experiments on microfluidic devices constructed in Cal Poly’s Microfabrication Laboratory. Although substantially functional, several design constraints had not yet been met. Our team has improved microscope robustness and functionality for practical undergraduate lab use. To do this, we set overarching goals including decreasing microscope footprint, increasing the accuracy of microscope positional repeatability, and improving user-friendliness. Within this Final Design Review report, the full design, manufacturing, and testing processes of this project are explicitly detailed, as well as project logistics, future suggestions, and project management

    Effect of Music Interventions on Sedation in Children Undergoing Magnetic Resonance Imaging: Clinical Trial

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    Background: Although parenteral sedation is often required in MRI studies in children, it is stressful and increases the cost of healthcare. Objectives: We evaluated the impact of music interventions in children receiving parenteral sedation for MRI studies on total number of doses of sedation medications, sedation time, levels of cortisol and cytokines, sedation success, adverse events, parental satisfaction, and cost savings. Methods: We conducted a prospective open unblinded four-arm clinical evaluation of interventions on 471 children 1-12 years of age undergoing MRI and receiving parenteral sedation. Children were assigned to active music therapy (AMT), facilitated music listening (FML), and as comparison another intervention (child life intervention or CLI), or no intervention (NI); measures included number of doses of sedation medications, time of sedation, sedation success, adverse events, parental satisfaction, and salivary levels of the stress hormone cortisol and pro-inflammatory cytokines, before and after intervention. Results: The total number of sedation doses, total sedation time, and levels of salivary cortisol and cytokines did not differ between the four groups. One FMLA choice, Wee Sing Animal Songs, resulted in significant decrease in total sedation time and reduction of associated costs. Conclusions: The use of one type of FML led to decreased total sedation time in children. This is an important finding since FML is an inexpensive non-invasive intervention which could be of significant time and cost saving benefits

    Kinetochore assembly and heterochromatin formation occur autonomously in Schizosaccharomyces pombe

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    Kinetochores in multicellular eukaryotes are usually associated with heterochromatin. Whether this heterochromatin simply promotes the cohesion necessary for accurate chromosome segregation at cell division or whether it also has a role in kinetochore assembly is unclear. Schizosaccharomyces pombe is an important experimental system for investigating centromere function, but all of the previous work with this species has exploited a single strain or its derivatives. The laboratory strain and most other S. pombe strains contain three chromosomes, but one recently discovered strain, CBS 2777, contains four. We show that the genome of CBS 2777 is related to that of the laboratory strain by a complex chromosome rearrangement. As a result, two of the kinetochores in CBS 2777 contain the central core sequences present in the laboratory strain centromeres, but lack adjacent heterochromatin. The closest block of heterochromatin to these rearranged kinetochores is ∼100 kb away at new telomeres. Despite lacking large amounts of adjacent heterochromatin, the rearranged kinetochores bind CENP-ACnp1 and CENP-CCnp3 in similar quantities and with similar specificities as those of the laboratory strain. The simplest interpretation of this result is that constitutive kinetochore assembly and heterochromatin formation occur autonomously

    Two spatial scales in a bleaching event : corals from the mildest and the most extreme thermal environments escape mortality

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    Author Posting. © Association for the Sciences of Limnology and Oceanography, 2013. This article is posted here by permission of Association for the Sciences of Limnology and Oceanography for personal use, not for redistribution. The definitive version was published in Limnology and Oceanography 58 (2013): 1531-1545, doi:10.4319/lo.2013.58.5.1531.In summer 2010, a bleaching event decimated the abundant reef flat coral Stylophora pistillata in some areas of the central Red Sea, where a series of coral reefs 100–300 m wide by several kilometers long extends from the coastline to about 20 km offshore. Mortality of corals along the exposed and protected sides of inner (inshore) and mid and outer (offshore) reefs and in situ and satellite sea surface temperatures (SSTs) revealed that the variability in the mortality event corresponded to two spatial scales of temperature variability: 300 m across the reef flat and 20 km across a series of reefs. However, the relationship between coral mortality and habitat thermal severity was opposite at the two scales. SSTs in summer 2010 were similar or increased modestly (0.5°C) in the outer and mid reefs relative to 2009. In the inner reef, 2010 temperatures were 1.4°C above the 2009 seasonal maximum for several weeks. We detected little or no coral mortality in mid and outer reefs. In the inner reef, mortality depended on exposure. Within the inner reef, mortality was modest on the protected (shoreward) side, the most severe thermal environment, with highest overall mean and maximum temperatures. In contrast, acute mortality was observed in the exposed (seaward) side, where temperature fluctuations and upper water temperature values were relatively less extreme. Refuges to thermally induced coral bleaching may include sites where extreme, high-frequency thermal variability may select for coral holobionts preadapted to, and physiologically condition corals to withstand, regional increases in water temperature.J.C.B.S. was partially supported by Fundac¸a˜o para a Cieˆncia e a Tecnologia (project PEst-C/MAR/LA0015/2011) and by the European Regional Development Fund through the Operational Competitiveness Programme (National Strategic Reference Framework). Kristen Davis was partially supported by a Woods Hole Oceanographic Institution postdoctoral scholarship. This research was supported by KAUST with awards USA 00002 and KSA 00011

    Mapping forest cover and forest cover change with airborne S-band radar

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    Assessments of forest cover, forest carbon stocks and carbon emissions from deforestation and degradation are increasingly important components of sustainable resource management, for combating biodiversity loss and in climate mitigation policies. Satellite remote sensing provides the only means for mapping global forest cover regularly. However, forest classification with optical data is limited by its insensitivity to three-dimensional canopy structure and cloud cover obscuring many forest regions. Synthetic Aperture Radar (SAR) sensors are increasingly being used to mitigate these problems, mainly in the L-, C- and X-band domains of the electromagnetic spectrum. S-band has not been systematically studied for this purpose. In anticipation of the British built NovaSAR-S satellite mission, this study evaluates the benefits of polarimetric S-band SAR for forest characterisation. The Michigan Microwave Canopy Scattering (MIMICS-I) radiative transfer model is utilised to understand the scattering mechanisms in forest canopies at S-band. The MIMICS-I model reveals strong S-band backscatter sensitivity to the forest canopy in comparison to soil characteristics across all polarisations and incidence angles. Airborne S-band SAR imagery over the temperate mixed forest of Savernake Forest in southern England is analysed for its information content. Based on the modelling results, S-band HH- and VV-polarisation radar backscatter and the Radar Forest Degradation Index (RFDI) are used in a forest/non-forest Maximum Likelihood classification at a spatial resolution of 6 m (70% overall accuracy, κ = 0.41) and 20 m (63% overall accuracy, κ = 0.27). The conclusion is that S-band SAR such as from NovaSAR-S is likely to be suitable for monitoring forest cover and its change
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