Evaluation of bacterial sampling methods for use with the bacterial tag-encoded flexible (FLX) amplicon pyrosequencing (bTEFAP) technique

2012 Summer.Includes bibliographical references.Background: The need to enumerate airborne microorganisms during infectious disease outbreaks, indoor air quality evaluations, and agricultural health studies has identified limitations in culture-based or viable sampling and characterization of bioaerosols. Pyrosequencing promises to be a novel, molecular-based technology that is exceptionally sensitive, low-cost, and provides a reasonable turnaround in the identification, distribution and concentration of aerosolized microorganisms. However, bioaerosol sampling methods for use with pyrosequencing have not been thoroughly evaluated. The intent of this project was to investigate a standardized sampling protocol for use with bTEFAP that would ultimately provide occupational scientists a novel and effective tool in the quest to characterize bioaerosol exposure and its subsequent relationship to worker health. Methods: Four filter types (Millipore Durapore® Membrane Filter, SKC water-soluble gelatin filter, SKC PTFE, SKC PVC) were prescreened for low-background DNA content using Pyrosequencing. Studies comparing the performance of the SKC Polyvinylchloride (PVC) and SKC gelatin filters in IOM samplers to an impinger - the SKC biosampler - were conducted in a previously characterized bioaerosol chamber using a Collision nebulizer. The challenge organism was a spore former, Bacillus atrophaeus. Tag-encoded FLX amplicon pyrosequencing analyses utilized Roche 454 FLX instrument with DNA extraction, massively parallel bTEFAP and bacterial identification data analysis was performed at the Research and Testing Laboratory (Lubbock, TX). Results: From an initial filter analysis, both the SKC PVC and SKC gelatin filters were selected for use in this project based on low-background DNA content, ease of use and cost. The two filter types and the SKC biosampler were challenged against B. atrophaeus for 30 minute sampling times in a series of six trials. Post pyrosequencing of detectable samples, it was demonstrated that the biosampler performed less effectively when compared to the PVC (p=0.0002) and gelatin filter (p=0.0006) based on an alpha value of 0.05. No significant difference was demonstrated between the two filter types (p=0.8). Of the original n=66 samples analyzed through pyrosequencing, only n=15 were reported to have counts for the challenge organism. In comparison to the pyrosequencing data, the cultured count demonstrated a significant difference when compared to the filters and biosampler media (p=0.003) in countable spores. Conclusions: The results indicate that with the model used in this study, the biosampler performed significantly different when compared to two filter types, the SKC PVC and the SKC gelatin, when challenged with B. atrophaeus. In addition, the microbial results suggest that there is possible significant contamination in the pyrosequencing methods used and or in the handling methods prior to analysis. Method analysis needs to be completed before further studies are completed

At My Best 4-5-6: Users’ observations of a health education curriculum-support resource

Background and context: At My Best 4-5-6 is a curriculum-support resource intended primarily for elementary generalist or physical and health education specialist teachers in Canada. Created by Physical and Health Education Canada and AstraZeneca, At My Best 4-5-6 aims to educate students about—and encourage healthy behaviours related to—physical activity, healthy eating, and emotional wellness. Objective: At My Best 4-5-6 is a relatively new program that is now being subject to a pilot-revision-pilot process. Given the lack of research literature focused upon the first iteration’s implementation, the objective was to complete an investigation of teachers’ observations related to the resource. Method: A cross-sectional survey design utilized both quantitative and qualitative data collected through on-line surveys. The data gleaned from 66 survey questions were analysed to explore program implementation and effectiveness, as well as overall teacher opinions regarding At My Best 4-5-6. Results and conclusion: We share results focused upon the following: 1) demographics: users, locations, schools, and usage, 2) need for and suitability of At My Best 4-5-6, 3) implementation, and 4) cultural relevance. Given these results, we offer some discussion and concluding observations related to present and future versions of At My Best 4-5-6. Moreover, we suggest such concluding observations might be of particular interest to others engaged in the creation, dissemination, and/or research of similar health education curriculum-support resources.Historique et contexte: À mon meilleur 4-5-6 est une ressource conçue comme soutien au programme d’études à l’élémentaire et visant surtout les enseignants généralistes ou les enseignants spécialistes en éducation physique ou en éducation à la santé au Canada. Créé par Éducation physique et santé Canada et AstraZeneca, À mon meilleur 4-5-6 a pour objectif de conscientiser les élèves quant à l’activité physique, une saine alimentation et le bienêtre émotionnel. Objectif : À mon meilleur 4-5-6 est un programme relativement récent qui fait maintenant l’objet d’un processus d’essai de modification. Compte tenu du manque de documents de recherche portant sur la mise en œuvre de la première version du programme, notre objectif était d’enquêter sur les observations des enseignants relatives à la ressource. Méthode: Une étude transversale a porté sur des données quantitatives et qualitatives provenant d’enquêtes enligne. Les données tirées de 66 questions d’enquête ont servi dans l’analyse de la mise en œuvre et l’efficacité du programme, ainsi que les opinions globales qu’ont les enseignants de cette ressource. Résultats et conclusion : Nos conclusions portent sur les quatre points suivants : 1) des données démographiques : usagers, emplacements, écoles et usage, 2) la nécessité et la pertinence de À mon meilleur 4-5-6, 3) la mise en œuvre, et 4) la pertinence culturelle. En nous appuyant sur ces résultats, nous offrons des éléments de discussion et des observations finales en lien avec les versions actuelle et future de À mon meilleur 4-5-6. Nous proposons, de plus, que de telles observations finales pourraient intéresser tout particulièrement les personnes impliquées dans la création, la diffusion ou la recherche de telles ressources conçues comme soutien au programme d’éducation à la santé.

Veterinarian nominated common conditions of rabbits and guinea pigs compared with published literature

Rabbits and guinea pigs are increasingly popular pets in the UK, yet little is known about their common ailments, or how these relate to what appears in the published literature. The aim of this study was to characterise the common conditions of rabbits and guinea pigs, and to compare these with the topics found in the published literature. Information about the common conditions seen in rabbits and guinea pigs in clinical practice was obtained from a survey of UK veterinarians. The common conditions seen were compared with results from a structured literature search. Conditions relating to the dental (29.9%), and skin (37.6%) body systems were commonly nominated by veterinarians for rabbits and guinea pigs, respectively. A total of 655 rabbit and 1086 guinea pig citations were examined and there appeared to be a mismatch between the conditions nominated in the veterinary questionnaire, and those found in the literature. This is the first time that the published literature has been compared to the nominated caseload of veterinarians in practice, and there is concern that the literature about rabbits and guinea pigs may not be representative of, or relevant to the caseload seen in clinical practice. This is of importance for clinicians being able to apply an objective, evidence-based approach. The publishing of clinically-relevant, research-based evidence should be prioritised

Forecasting Innovation Pathways (FIP) for new and emerging science and technologies

International audience"New" and "Emerging Science" and "Technologies" ("NESTs") have tremendous innovation potential. However this must be weighed against enormous uncertainties caused by many unknowns. The authors of this paper offer a framework to analyze NESTs to help ascertain likely innovation pathways.We have devised a 10-step framework based on extensive Future-oriented Technology Analyses ("FTA") experience, enriched by in-depth case analyses. In the paper, we describe our analytical activities in two case studies. The nanobiosensor experience is contrasted with that of deep brain stimulation in relative quantitative and qualitative emphases.We close the paper by reflecting on this systematic FTA framework for emerging science and technologies, for its intended goal, that is to support decision making

Functional Plasticity in Lamellar Autotomy by Larval Damselflies in Response to Predatory Larval Dragonfly Cues

Adaptive autotomy is the self-amputation of an appendage in response to external stimuli that benefits survival. Variation in the ease of appendage removal among populations suggests that autotomy performance is under selection, evolves, or is phenotypically plastic, although the latter has never been experimentally tested. We model an autotomy threshold that optimally balances how the benefits of surviving predator attack versus the costs of losing an appendage vary with predator presence. We test for functional plasticity in autotomy threshold in the caudal lamellae of Enallagma damselfly larvae by experimentally manipulating non-lethal cues from predatory dragonfly larvae. Predator cues lead to functional plastic responses in the form of smaller lamellar joints that required lower peak breaking force. This is the first experimental demonstration of functional plasticity in autotomy to cues from a grasping predator, a novel form of indirect predator effects on prey, realized through plasticity in morphological traits that govern the autotomy threshold. This supports the model of optimized autotomy performance and provides a novel explanation for variation in performance among populations under different predator conditions. Plastic autotomy responses that mitigate costs in the face of variation in mortality risks might be a form of inducible defense

Using Gravity Recovery and Climate Experiment data to derive corrections to precipitation data sets and improve modelled snow mass at high latitudes

The amount of lying snow calculated by a land surface model depends in part on the amount of snowfall in the meteorological data that are used to drive the model. We show that commonly used data sets differ in the amount of snowfall, and more generally precipitation, over four large Arctic basins. An independent estimate of the cold-season precipitation is obtained by combining water balance information from the Gravity Recovery and Climate Experiment (GRACE) with estimates of evaporation and river discharge and is generally higher than that estimated by four commonly used meteorological data sets. We use the Joint UK Land Environment Simulator (JULES) land surface model to calculate the snow water equivalent (SWE) over the four basins. The modelled seasonal maximum SWE is 38 % less than observation-based estimates on average, and the modelled basin discharge is significantly underestimated, consistent with the lack of snowfall. We use the GRACE-derived estimate of precipitation to define per-basin scale factors that are applied to the driving data and increase the amount of cold-season precipitation by 28 % on average. In turn this increases the modelled seasonal maximum SWE by 30 %, although this is still underestimated compared to observations by 19 % on average. A correction for the undercatch of precipitation by gauges is compared with the the GRACE-derived correction. Undercatch correction increases the amount of cold-season precipitation by 23 % on average, which indicates that some, but not all, of the underestimation can be removed by implementing existing undercatch correction algorithms. However, even undercatch-corrected data sets contain less precipitation than the GRACE-derived estimate in some regions, and it is likely that there are other biases that are not currently accounted for in gridded meteorological data sets. This study shows that revised estimates of precipitation can lead to improved modelling of SWE, but much more modest improvements are found in modelled river discharge. By providing methods to better define the precipitation inputs to the system, the current study paves the way for subsequent work on key hydrological processes in high-latitude basins

Trends in atmospheric evaporative demand in Great Britain using high-resolution meteorological data

Observations of climate are often available on very different spatial scales from observations of the natural environments and resources that are affected by climate change. In order to help bridge the gap between these scales using modelling, a new dataset of daily meteorological variables was created at 1 km resolution over Great Britain for the years 1961–2012, by interpolating coarser resolution climate data and including the effects of local topography. These variables were used to calculate atmospheric evaporative demand (AED) at the same spatial and temporal resolution. Two functions that represent AED were chosen: one is a standard form of potential evapotranspiration (PET) and the other is a derived PET measure used by hydrologists that includes the effect of water intercepted by the canopy (PETI). Temporal trends in these functions were calculated, with PET found to be increasing in all regions, and at an overall rate of 0.021 ± 0.021 mm day−1 decade−1 in Great Britain. PETI was found to be increasing at a rate of 0.019 ± 0.020 mm day−1 decade−1 in Great Britain, but this was not statistically significant. However, there was a trend in PETI in England of 0.023 ± 0.023 mm day−1 decade−1. The trends were found to vary by season, with spring PET increasing by 0.043 ± 0.019 mm day−1 decade−1 (0.038 ± 0.018 mm day−1 decade−1 when the interception correction is included) in Great Britain, while there is no statistically significant trend in other seasons. The trends were attributed analytically to trends in the climate variables; the overall positive trend was predominantly driven by rising air temperature, although rising specific humidity had a negative effect on the trend. Recasting the analysis in terms of relative humidity revealed that the overall effect is that falling relative humidity causes the PET to rise. Increasing downward short- and longwave radiation made an overall positive contribution to the PET trend, while decreasing wind speed made a negative contribution to the trend in PET. The trend in spring PET was particularly strong due to a strong decrease in relative humidity and increase in downward shortwave radiation in the spring

Exploitation of the Escherichia coli lac operon promoter for controlled recombinant protein production

The Escherichia coli lac operon promoter is widely used as a tool to control recombinant protein production in bacteria. Here we give a brief review of how it functions, how it is regulated, and how, based on this knowledge, a suite of lac promoter derivatives has been developed to give controlled expression that is suitable for diverse biotechnology applications

The Direct Medical Costs of Undiagnosed Chronic Obstructive Pulmonary Disease

AbstractObjectiveTo estimate the costs of undiagnosed chronic obstructive pulmonary disease (COPD) by describing inpatient, outpatient, and pharmacy utilization in the years before and after the diagnosis.MethodsA total of 6864 patients who were enrolled in the Lovelace Health Plan for at least 12 months during the study period (January 1, 1999 through December 31, 2004) were identified. The first date that utilization was attributed to COPD was considered the first date of diagnosis. Each COPD case was matched to up to three age- and sex-matched controls. All utilization and direct medical costs during the study period were compiled monthly and compared based on the time before and after the initial diagnosis.ResultsTotal costs were higher by an average of $1182 per patient in the 2 years before the initial COPD diagnosis, and$2489 in the 12 months just before the initial diagnosis, compared to matched controls. Most of the higher cost for undiagnosed COPD was attributable to hospitalizations. Inpatient costs did not increase after the diagnosis was made, but approximately one-third of admissions after the diagnosis were attributed to respiratory disease. Outpatient and pharmacy costs did not differ substantially between cases and matched controls until just a few months before the initial diagnosis, but remained 50% to 100% higher than for controls in the 2 years after diagnosis.ConclusionsUndiagnosed COPD has a substantial impact on health-care costs and utilization in this integrated managed care system, particularly for hospitalizations