Evaluation of the environmental specificity of fluorescence in situ hybridization (FISH) using fluorescence-activated cell sorting (FACS) of probe (PSE1284)-positive cells extracted from rhizosphere soil

We explicitly tested for the first time the ‘environmental specificity’ of traditional 16S rRNAtargeted fluorescence in situ hybridization (FISH) through comparison of the bacterial diversity actually targeted in the environment with the diversity that should be exactly targeted (i.e. without mismatches) according to in silico analysis. To do this, we exploited advances in modern Flow Cytometry that enabled improved detection and therefore sorting of sub-micron-sized particles and used probe PSE1284 (designed to target Pseudomonads) applied to Lolium perenne rhizosphere soil as our test system. The 6-carboxyfluorescein (6-FAM)-PSE1284-hybridised population, defined as displaying enhanced green fluorescence in Flow Cytometry, represented 3.51±1.28% of the total detected population when corrected using a nonsense (NON-EUB338) probe control. Analysis of 16S rRNA gene libraries constructed from Fluorescence Activated Cell Sorted (FACS) -recovered fluorescent populations (n=3), revealed that 98.5% (Pseudomonas spp. comprised 68.7% and Burkholderia spp. 29.8%) of the total sorted population was specifically targeted as evidenced by the homology of the 16S rRNA sequences to the probe sequence. In silico evaluation of probe PSE1284 with the use of RDP-10 probeMatch justified the existence of Burkholderia spp. among the sorted cells. The lack of novelty in Pseudomonas spp. sequences uncovered was notable, probably reflecting the well-studied nature of this functionally important genus. To judge the diversity recorded within the FACS-sorted population, rarefaction and DGGE analysis were used to evaluate, respectively, the proportion of Pseudomonas diversity uncovered by the sequencing effort and the representativeness of the Nycodenz® method for the extraction of bacterial cells from soil.

Emotional well-being and adjustment to vision loss in later life: a meta-synthesis of qualitative studies.

Purpose: To review perceived emotional well-being in older people with visual impairment and perceived factors that inhibit/facilitate psychosocial adjustment to vision loss. Method: The databases of MEDLINE, EMBASE, PsycINFO and CINAHL were searched for studies published from January 1980 to December 2010, which recruited older people with irreversible vision loss, and used qualitative methods for both data collection and analysis. Results sections of the papers were synthesised using a thematic-style analysis to identify the emergent and dominant themes. Results: Seventeen qualitative papers were included in the review, and five main themes emerged from the synthesis: 1) the trauma of an ophthalmic diagnosis, 2) impact of vision loss on daily life, 3) negative impact of visual impairment on psychosocial well-being, 4) factors that inhibit social well-being, and 5) factors that facilitate psychological well-being. We found the response shift model useful for explaining our synthesis. Conclusions: Acquired visual impairment can have a significant impact on older people's well-being and make psychosocial adjustment to the condition a major challenge. Acceptance of the condition and a positive attitude facilitate successful psychosocial adjustment to vision loss as well as social support from family, friends and peers who have successfully adjusted to the condition. [Box: see text].

Evaluation of field wetlands for mitigation of diffuse pollution from agriculture: sediment retention, cost and effectiveness

Diffuse pollution, and the contribution from agriculture in particular, has become increasingly important as pollution from point sources has been addressed by wastewater treatment. Land management approaches, such as construction of field wetlands, provide one group of mitigation options available to farmers. Although field wetlands are widely used for diffuse pollution control in temperate environments worldwide, there is a shortage of evidence for the effectiveness and viability of these mitigation options in the UK. The Mitigation Options for Phosphorus and Sediment Project aims to make recommendations regarding the design and effectiveness of field wetlands for diffuse pollution control in UK landscapes. Ten wetlands have been built on four farms in Cumbria and Leicestershire. This paper focuses on sediment retention within the wetlands, estimated from annual sediment surveys in the first two years, and discusses establishment costs. It is clear that the wetlands are effective in trapping a substantial amount of sediment. Estimates of annual sediment retention suggest higher trapping rates at sandy sites (0.5–6 t ha�1 yr�1), compared to silty sites (0.02–0.4 t ha�1 yr�1) and clay sites (0.01–0.07 t ha�1 yr�1). Establishment costs for the wetlands ranged from £280 to £3100 and depended more on site specific factors, such as fencing and gateways on livestock farms, rather than on wetland size or design. Wetlands with lower trapping rates would also have lower maintenance costs, as dredging would be required less frequently. The results indicate that field wetlands show promise for inclusion in agri-environment schemes, particularly if capital payments can be provided for establishment, to encourage uptake of these multi-functional features.

The impact of social and relational contexts on innovation transfer in foreign subsidiaries

It is widely acknowledged that innovation is one of the pillars of multinational enterprises (MNEs) and that technological knowledge from different host locations is a key factor to the MNEs’ competitive advantages development. Concerning these assumptions, in this paper we aim to understand how the social and the relational contexts affect the conventional and reverse transfer of innovation from MNEs’ subsidiaries hosted in emerging markets. We analyzed the social context through the institutional profile (CIP) level and the relational context through trust and integration levels utilizing a survey sent to 172 foreign subsidiaries located in Brazil, as well as secondary data. Through an ordinary least squares regression (OLS) analysis we found that the relational context affects the conventional and reverse innovation transfer in subsidiaries hosted in emerging markets. We however did not find support for the social context effect.

Social citizenship, housing wealth and the cost of social care: is the Care Act 2014 'Fair'?

This article assesses the extent to which it is ‘fair’ for the government to require owner-occupiers to draw on the equity accumulated in their home to fund their social care costs. The question is stimulated by the report of the Commission on Funding of Care and Support, Fairer Care Funding (the Dilnot Commission) and the subsequent Care Act 2014. The enquiry is located within the framework of social citizenship and the new social contract. It argues that the individualistic, contractarian approach, exemplified by the Dilnot Commission and reflected in the Act, raises questions when considered from the perspective of intergenerational fairness. We argue that our concerns with the Act could be addressed by inculcating an expectation of drawing on housing wealth to fund older age: a policy of asset-based welfare.

An evaluation of the theoretical potential and practical opportunity for using greywater for domestic purposes in Ghana

Assessments concerning the effects of climate change, water resource availability and water deprivation in West Africa have not frequently considered the positive contribution to be derived from collecting and reusing water for domestic purposes. Where the originating water is taken from a clean water source and has been used the first time for washing or bathing, this water is commonly called “greywater”. Greywater is a prolific resource that is generated wherever people live. Treated greywater can be used for domestic cleaning, for flushing toilets where appropriate, for washing cars, sometimes for watering kitchen gardens, and for clothes washing prior to rinsing. Therefore, a large theoretical potential exists to increase total water resource availability if greywater were to be widely reused. Locally treated greywater reduces the distribution network requirement, lower construction effort and cost and, wherever possible, minimising the associated carbon footprint. Such locally treated greywater offers significant practical opportunities for increasing the total available water resources at a local level. The reuse of treated greywater is one important action that will help to mitigate the reducing availability of clean water supplies in some areas, and the expected mitigation required in future aligns well with WHO/UNICEF (2012) aspirations. The evaluation of potential opportunities for prioritising greywater systems to support water reuse takes into account the availability of water resources, water use indicators and published estimates in order to understand typical patterns of water demand. The approach supports knowledge acquisition regarding local conditions for enabling capacity building for greywater reuse, the understanding of systems that are most likely to encourage greywater reuse, and practices and future actions to stimulate greywater infrastructure planning, design and implementation. Although reuse might be considered to increase the uncertainty of achieving a specified quality of the water supply, robust methods and technologies are available for local treatment. Resource strategies for greywater reuse have the potential to consistently improve water efficiency and availability in water impoverished and water stressed regions of Ghana and West Africa. Untreated greywater is referred to as “greywater”; treated greywater is referred to as “treated greywater” in this paper.

Synthesis and evaluation of a novel hydrophilic 6,6′-bis(1,2,4-triazin-3-yl)-2,2′-bipyridine ligand for separating actinide(III) from lanthanide(III)

We report the synthesis and evaluation of a novel hydrophilic 6,6′-bis(1,2,4-triazin-3-yl)-2,2′-bipyridine (BTBP) ligand containing carboxylate groups as a selective aqueous complexing agent for the minor actinides over lanthanides. The novel ligand is able to complex and separate Am(III) from Eu(III) in alkaline solutions selectively.

Evaluation of the realism of climate reconstruction using the Coexistence Approach with modern pollen samples from the Tibetan Plateau

We apply the Coexistence Approach (CoA) to reconstruct mean annual precipitation (MAP), mean annual temperature (MAT), mean temperature of thewarmestmonth (MTWA) and mean temperature of the coldest month (MTCO) at 44 pollen sites on the Qinghai–Tibetan Plateau. The modern climate ranges of the taxa are obtained (1) from county-level presence/absence data and (2) from data on the optimum and range of each taxon from Lu et al. (2011). The CoA based on the optimumand range data yields better predictions of observed climate parameters at the pollen sites than that based on the county-level data. The presence of arboreal pollen, most of which is derived fromoutside the region, distorts the reconstructions. More reliable reconstructions are obtained using only the non-arboreal component of the pollen assemblages. The root mean-squared error (RMSE) of the MAP reconstructions are smaller than the RMSE of MAT, MTWA and MTCO, suggesting that precipitation gradients are the most important control of vegetation distribution on the Qinghai–Tibetan Plateau. Our results show that CoA could be used to reconstruct past climates in this region, although in areas characterized by open vegetation the most reliable estimates will be obtained by excluding possible arboreal contaminants.

Evaluation of a modern-analogue methodology for reconstructing Australian palaeoclimate from pollen

Quantitative palaeoclimate reconstructions are widely used to evaluate climatemodel performance. Here, as part of an effort to provide such a data set for Australia, we examine the impact of analytical decisions and sampling assumptions on modern-analogue reconstructions using a continent-wide pollen data set. There is a high degree of correlation between temperature variables in the modern climate of Australia, but there is sufficient orthogonality in the variations of precipitation, summer and winter temperature and plant–available moisture to allow independent reconstructions of these four variables to be made. The method of analogue selection does not affect the reconstructions, although bootstrap resampling provides a more reliable technique for obtaining robust measures of uncertainty. The number of analogues used affects the quality of the reconstructions: the most robust reconstructions are obtained using 5 analogues. The quality of reconstructions based on post-1850 CE pollen samples differ little from those using samples from between 1450 and 1849 CE, showing that European post settlement modification of vegetation has no impact on the fidelity of the reconstructions although it substantially increases the availability of potential analogues. Reconstructions based on core top samples are more realistic than those using surface samples, but only using core top samples would substantially reduce the number of available analogues and therefore increases the uncertainty of the reconstructions. Spatial and/or temporal averaging of pollen assemblages prior to analysis negatively affects the subsequent reconstructions for some variables and increases the associated uncertainties. In addition, the quality of the reconstructions is affected by the degree of spatial smoothing of the original climate data, with the best reconstructions obtained using climate data froma 0.5° resolution grid, which corresponds to the typical size of the pollen catchment. This study provides a methodology that can be used to provide reliable palaeoclimate reconstructions for Australia, which will fill in a major gap in the data sets used to evaluate climate models.

Evaluation of simulated sea-ice concentrations from sea-ice/ocean models using satellite data and polynya classification methods

Sea-ice concentrations in the Laptev Sea simulated by the coupled North Atlantic-Arctic Ocean-Sea-Ice Model and Finite Element Sea-Ice Ocean Model are evaluated using sea-ice concentrations from Advanced Microwave Scanning Radiometer-Earth Observing System satellite data and a polynya classification method for winter 2007/08. While developed to simulate largescale sea-ice conditions, both models are analysed here in terms of polynya simulation. The main modification of both models in this study is the implementation of a landfast-ice mask. Simulated sea-ice fields from different model runs are compared with emphasis placed on the impact of this prescribed landfast-ice mask. We demonstrate that sea-ice models are not able to simulate flaw polynyas realistically when used without fast-ice description. Our investigations indicate that without landfast ice and with coarse horizontal resolution the models overestimate the fraction of open water in the polynya. This is not because a realistic polynya appears but due to a larger-scale reduction of ice concentrations and smoothed ice-concentration fields. After implementation of a landfast-ice mask, the polynya location is realistically simulated but the total open-water area is still overestimated in most cases. The study shows that the fast-ice parameterization is essential for model improvements. However, further improvements are necessary in order to progress from the simulation of large-scale features in the Arctic towards a more detailed simulation of smaller-scaled features (here polynyas) in an Arctic shelf sea.

Evaluation of the pH and electrical conductivity in milk from jersey cows during the first month of lactation

In order to assess the influence of the colostrum period on pH and, electrical conductivity, we collected 418 milk samples from 127 Jersey cows. The samples were collected from healthy udders that did not present any bacterial growth in the microbiological examination. They were divided into eight groups as follows < 1/2 day; 1/2 and 1 degrees day; 2 degrees day; 3 degrees day; 4 degrees and 5 degrees day; 6 degrees and 7 degrees day; 8 degrees to 15 degrees day; 16 degrees to 30 degrees days of lactation. The samples were collected before milking and the following analyses were conducted: pH, electrical conductivity. In the first 24 hours of lactation, there was an reduction in electrical conductivity value, associated with an increase in pH value. We observed that transition of secretion from colostrum to milk, occurs during the first week of lactation; from 6(rd) day of lactation for pH value and 3(th) day for electrical conductivity value. We recommend the use the following figures as normal ranges for the first 24 hours of lactation (colostrum period): pH <= 6,51 and electrical conductivity <= 6,33 mS/cm; while for the interval between 2(nd) and 7(th) days of lactation (transition from colostrum to milk) we suggest the use of the values as normal ranges: pH <= 6,66 and electrical conductivity <= 5,93 mS/cm.

Research on Candida dubliniensis in a Brazilian yeast collection obtained from cardiac transplant, tuberculosis, and HIV-positive patients, and evaluation of phenotypic tests using agar screening methods

The aim of this study was to research Candida dubliniensis among isolates present in a Brazilian yeast collection and to evaluate the main phenotypic methods for discrimination between C. albicans and C. dubliniensis from oral cavity. A total of 200 isolates, presumptively identified as C. albicans or C. dubliniensis obtained from heart transplant patients under immunosuppressive therapy, tuberculosis patients under antibiotic therapy, HIV-positive patients under antiretroviral therapy, and healthy subjects, were analyzed using the following phenotypic tests: formation and structural arrangement of chlamydospores on corn meal agar, casein agar, tobacco agar, and sunflower seed agar; growth at 45 degrees C; and germ tube formation. All strains were analyzed by polymerase chain reaction (PCR). In a preliminary screen for C. dubliniensis, 48 of the 200 isolates on corn meal agar, 30 of the 200 on casein agar, 16 of the 200 on tobacco agar, and 15 of the 200 on sunflower seed agar produced chlamydoconidia; 27 of the 200 isolates showed no or poor growth at 45 degrees C. All isolates were positive for germ tube formation. These isolates were considered suggestive of C. dubliniensis. All of them were subjected to PCR analysis using C. dubliniensis-specific primers. C. dubliniensis isolates were not found. C. dubliniensis isolates were not recovered in this study done with immunocompromised patients. Sunflower seed agar was the medium with the smallest number of isolates of C. albicans suggestive of C. dubliniensis. None of the phenotypic methods was 100% effective for discrimination between C. albicans and C. dubliniensis. (C) 2011 Elsevier Inc. All rights reserved.

Inconclusive results in conventional serological screening for Chagas` disease in blood banks: evaluation of cellular and humoral response

To find the most reliable screening method for Trypanosoma cruzi infection in blood banks. Epidemiological data, lymphoproliferation assay, parasitological, conventional serological tests: immunofluorescence, haemagglutination, ELISA with epimastigote and trypomastigote antigens and reference serological tests: trypomastigote excreted-secreted antigens (TESA) blot and chemiluminescent ELISA assay with mucine from trypomastigote forms were applied to individuals with inconclusive serology, non-chagasic individuals and chronic chagasic patients. TESA blot had the best performance when used as a single test in all the groups. In the inconclusive group 20.5% of individuals were positive for TESA blot, 23.3% for either lymphoproliferation or TESA blot, and 17.8% for lymphoproliferation only. Positive lymphoproliferation without detectable antibodies was observed in 5.47% of all inconclusive serology cases. Analysis of six parameters (three serological assays, at least one parasitological test, one lymphoproliferation assay and epidemiological data) in the inconclusive group showed that diagnosis of Chagas` disease was probable in 15 patients who were positive by two or more serological tests or for whom three of those six parameters were positive. TESA blot is a good confirmatory test for Chagas` disease in the inconclusive group. Although lymphoproliferation suggests the diagnosis of Chagas` disease in the absence of antibodies when associated with a high epidemiological risk of acquiring Chagas` disease, the data from this study and the characteristics of the lymphoproliferation assay (which is both laborious and time-consuming) do not support its use as a confirmatory test in blood-bank screening. However, our findings underscore the need to develop alternative methods that are not based on antibody detection to improve the diagnosis when serological tests are inconclusive.