Use of Rapid-Scan EPR to Improve Detection Sensitivity for Spin-Trapped Radicals

AbstractThe short lifetime of superoxide and the low rates of formation expected in vivo make detection by standard continuous wave (CW) electron paramagnetic resonance (EPR) challenging. The new rapid-scan EPR method offers improved sensitivity for these types of samples. In rapid-scan EPR, the magnetic field is scanned through resonance in a time that is short relative to electron spin relaxation times, and data are processed to obtain the absorption spectrum. To validate the application of rapid-scan EPR to spin trapping, superoxide was generated by the reaction of xanthine oxidase and hypoxanthine with rates of 0.1–6.0 μM/min and trapped with 5-tert-butoxycarbonyl-5-methyl-1-pyrroline-N-oxide (BMPO). Spin trapping with BMPO to form the BMPO-OOH adduct converts the very short-lived superoxide radical into a more stable spin adduct. There is good agreement between the hyperfine splitting parameters obtained for BMPO-OOH by CW and rapid-scan EPR. For the same signal acquisition time, the signal/noise ratio is >40 times higher for rapid-scan than for CW EPR. Rapid-scan EPR can detect superoxide produced by Enterococcus faecalis at rates that are too low for detection by CW EPR

COVID-19 and OD: Unplanned Disruption and the Opportunity for Planned Talent Development

COVID-19 created an unprecedented global health crisis and caused a rapid, global economic meltdown. Organizations of all sizes are scrambling to salvage workforces and adopting policies to protect employees from the virus. Talent development practices face new challenges as the world reckons with our altered COVID-19 pandemic reality. Organization leaders must think creatively to design the employment practices of the future. In this article, we consider how changes in work environments affect employees and organization performance, examine how recruitment, training, onboarding, and developing talent present complex challenges that must be addressed for sustainability, and consider how talent development principles can enhance OD practices. Finally, we offer perspectives on employing OD principles and practices to facilitate workforce changes to maximize productivity and performance

Sewage reflects the microbiomes of human populations

© The Author(s), 2015. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in mBio 6 (2015): e02574-14, doi:10.1128/mBio.02574-14.Molecular characterizations of the gut microbiome from individual human stool samples have identified community patterns that correlate with age, disease, diet, and other human characteristics, but resources for marker gene studies that consider microbiome trends among human populations scale with the number of individuals sampled from each population. As an alternative strategy for sampling populations, we examined whether sewage accurately reflects the microbial community of a mixture of stool samples. We used oligotyping of high-throughput 16S rRNA gene sequence data to compare the bacterial distribution in a stool data set to a sewage influent data set from 71 U.S. cities. On average, only 15% of sewage sample sequence reads were attributed to human fecal origin, but sewage recaptured most (97%) human fecal oligotypes. The most common oligotypes in stool matched the most common and abundant in sewage. After informatically separating sequences of human fecal origin, sewage samples exhibited ~3× greater diversity than stool samples. Comparisons among municipal sewage communities revealed the ubiquitous and abundant occurrence of 27 human fecal oligotypes, representing an apparent core set of organisms in U.S. populations. The fecal community variability among U.S. populations was significantly lower than among individuals. It clustered into three primary community structures distinguished by oligotypes from either: Bacteroidaceae, Prevotellaceae, or Lachnospiraceae/Ruminococcaceae. These distribution patterns reflected human population variation and predicted whether samples represented lean or obese populations with 81 to 89% accuracy. Our findings demonstrate that sewage represents the fecal microbial community of human populations and captures population-level traits of the human microbiome.Funding for this work was provided by the NIH grant R01AI091829-01A1 to S.L.M. and M.L.S

Unique Microbial Communities Persist in Individual Cystic Fibrosis Patients throughout a Clinical Exacerbation

Cystic fibrosis (CF) is caused by inherited mutations in the cystic fibrosis transmembrane conductance regulator gene and results in a lung environment that is highly conducive to polymicrobial infection. Over a lifetime, decreasing bacterial diversity and the presence of Pseudomonas aeruginosa in the lung are correlated with worsening lung disease. However, to date, no change in community diversity, overall microbial load or individual microbes has been shown to correlate with the onset of an acute exacerbation in CF patients. We followed 17 adult CF patients throughout the course of clinical exacerbation, treatment and recovery, using deep sequencing and quantitative PCR to characterize spontaneously expectorated sputum sample

Distributed deformation ahead of the Cocos-Nazca Rift at the Galapagos triple junction

Author Posting. © American Geophysical Union, 2011. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Geochemistry Geophysics Geosystems 12 (2011): Q11003, doi:10.1029/2011GC003689.The Galapagos triple junction is not a simple ridge-ridge-ridge (RRR) triple junction. The Cocos-Nazca Rift (C-N Rift) tip does not meet the East Pacific Rise (EPR). Instead, two secondary rifts form the link: Incipient Rift at 2°40′N and Dietz Deep volcanic ridge, the southern boundary of the Galapagos microplate (GMP), at 1°10′N. Recently collected bathymetry data are used to investigate the regional tectonics prior to the establishment of the GMP (∼1.5 Ma). South of C-N Rift a band of northeast-trending cracks cuts EPR-generated abyssal hills. It is a mirror image of a band of cracks previously identified north of C-N Rift on the same age crust. In both areas, the western ends of the cracks terminate against intact abyssal hills suggesting that each crack initiated at the EPR spreading center and cut eastward into pre-existing topography. Each crack formed a short-lived triple junction until it was abandoned and a new crack and triple junction initiated nearby. Between 2.5 and 1.5 Ma, the pattern of cracking is remarkably symmetric about C-N Rift providing support for a crack interaction model in which crack initiation at the EPR axis is controlled by stresses associated with the tip of the westward-propagating C-N Rift. The model also shows that offsets of the EPR axis may explain times when cracking is not symmetric. South of C-N Rift, cracks are observed on seafloor as old as 10.5 Ma suggesting that this triple junction has not been a simple RRR triple junction during that time.HS was supported by the U.S. National Science Foundation (NSF) grant OCE‐0751831, DS by NSF grant OCE‐1028537, WZ by NSF grant EAR‐1056317, and LM by NSF grant EAR‐0911151.2012-05-0

Using workplace population statistics to understand retail store performance

We explore the value of recently released workplace geographies and accompanying census-based workplace zone statistics (WZS) and an associated classification of workplace zones (COWZ). We consider how these data could support retailers in their operational and strategic decision making, including the evaluation of retail demand and retail store performance in localities where trade is driven by non-residential demand. In collaboration with major UK grocery retailer ‘The Co-operative Group’ we explore the relationship between workplace population composition and store trading characteristics using a series of case study stores within Inner London. We use empirical store trading data to identify store and product category level temporal sales fluctuations attributable to workplace populations. We also use census-derived flow data to identify the spatial origins of workplace population inflow. We identify that store performance exhibits characteristics attributable to demand driven by these populations. We conclude that workplace population geographies, WZS and the COWZ afford considerable potential for understanding drivers of store performance, observed store trading patterns and evaluation of retail store performance. We suggest that the next step is to build these populations and their micro geography spatial and temporal characteristics into predictive models and evaluate their potential for store performance evaluation and location-based store and network decision making within this sector

Caring for the patient, caring for the record: an ethnographic study of 'back office' work in upholding quality of care in general practice

© 2015 Swinglehurst and Greenhalgh; licensee BioMed Central. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.Additional file 1: Box 1. Field notes on summarising (Clover Surgery). Box 2. Extract of document prepared for GPs by summarisers at Clover Surgery. Box 3. Fieldnotes on coding incoming post, Clover (original notes edited for brevity).This work was funded by a research grant from the UK Medical Research Council (Healthcare Electronic Records in Organisations 07/133) and a National Institute of Health Research doctoral fellowship award for DS (RDA/03/07/076). The funders were not involved in the selection or analysis of data nor did they make any contribution to the content of the final manuscript

Characterization and quantification of the fungal microbiome in serial samples from individuals with cystic fibrosis

Abstract Background Human-associated microbial communities include fungi, but we understand little about which fungal species are present, their relative and absolute abundances, and how antimicrobial therapy impacts fungal communities. The disease cystic fibrosis (CF) often involves chronic airway colonization by bacteria and fungi, and these infections cause irreversible lung damage. Fungi are detected more frequently in CF sputum samples upon initiation of antimicrobial therapy, and several studies have implicated the detection of fungi in sputum with worse outcomes. Thus, a more complete understanding of fungi in CF is required. Results We characterized the fungi and bacteria in expectorated sputa from six CF subjects. Samples were collected upon admission for systemic antibacterial therapy and upon the completion of treatment and analyzed using a pyrosequencing-based analysis of fungal internal transcribed spacer 1 (ITS1) and bacterial 16S rDNA sequences. A mixture of Candida species and Malassezia dominated the mycobiome in all samples (74%–99% of fungal reads). There was not a striking trend correlating fungal and bacterial richness, and richness showed a decline after antibiotic therapy particularly for the bacteria. The fungal communities within a sputum sample resembled other samples from that subject despite the aggressive antibacterial therapy. Quantitative PCR analysis of fungal 18S rDNA sequences to assess fungal burden showed variation in fungal density in sputum before and after antibacterial therapy but no consistent directional trend. Analysis of Candida ITS1 sequences amplified from sputum or pure culture-derived genomic DNA from individual Candida species found little (<0.5%) or no variation in ITS1 sequences within or between strains, thereby validating this locus for the purpose of Candida species identification. We also report the enhancement of the publically available Visualization and Analysis of Microbial Population Structures (VAMPS) tool for the analysis of fungal communities in clinical samples. Conclusions Fungi are present in CF respiratory sputum. In CF, the use of intravenous antibiotic therapy often does not profoundly impact bacterial community structure, and we observed a similar stability in fungal species composition. Further studies are required to predict the effects of antibacterials on fungal burden in CF and fungal community stability in non-CF populations.http://deepblue.lib.umich.edu/bitstream/2027.42/134558/1/40168_2014_Article_67.pd

Characterization and Quantification of the Fungal Microbiome in Serial Samples from Individuals with Cystic Fibrosis

Background: Human-associated microbial communities include fungi, but we understand little about which fungal species are present, their relative and absolute abundances, and how antimicrobial therapy impacts fungal communities. The disease cystic fibrosis (CF) often involves chronic airway colonization by bacteria and fungi, and these infections cause irreversible lung damage. Fungi are detected more frequently in CF sputum samples upon initiation of antimicrobial therapy, and several studies have implicated the detection of fungi in sputum with worse outcomes. Thus, a more complete understanding of fungi in CF is required. Results: We characterized the fungi and bacteria in expectorated sputa from six CF subjects. Samples were collected upon admission for systemic antibacterial therapy and upon the completion of treatment and analyzed using a pyrosequencing-based analysis of fungal internal transcribed spacer 1 (ITS1) and bacterial 16S rDNA sequences. A mixture of Candida species and Malassezia dominated the mycobiome in all samples (74% – 99% of fungal reads). There was not a striking trend correlating fungal and bacterial richness, and richness showed a decline after antibiotic therapy particularly for the bacteria. The fungal communities within a sputum sample resembled other samples from that subject despite the aggressive antibacterial therapy. Quantitative PCR analysis of fungal 18S rDNA sequences to assess fungal burden showed variation in fungal density in sputum before and after antibacterial therapy but no consistent directional trend. Analysis of Candida ITS1 sequences amplified from sputum or pure culture-derived genomic DNA from individual Candida species found little (\u3c0.5%) or no variation in ITS1 sequences within or between strains, thereby validating this locus for the purpose of Candida species identification. We also report the enhancement of the publically available Visualization and Analysis of Microbial Population Structures (VAMPS) tool for the analysis of fungal communities in clinical samples. Conclusions: Fungi are present in CF respiratory sputum. In CF, the use of intravenous antibiotic therapy often does not profoundly impact bacterial community structure, and we observed a similar stability in fungal species composition. Further studies are required to predict the effects of antibacterials on fungal burden in CF and fungalcommunity stability in non-CF populations