bluuebunny/crossref_metadata_2025 huggingface dataset:
Improvement in Thermostability of an Achaetomium sp. Strain Xz8 Endopolygalacturonase via the Optimization of Charge-Charge Interactions
Authors: Tu Tao , Luo Huiying , Meng Kun , Cheng Yanli , Ma Rui , Shi Pengjun , Huang Huoqing , Bai Yingguo , Wang Yaru , Zhang Lujia , Yao Bin
Published: July 2015

Improving enzyme thermostability is of importance for widening the spectrum of application of enzymes. In this study, a structure-based rational design approach was used to improve the thermostability of a highly active, wide-pH-range-adaptable, and stable endopolygalacturonase (PG8fn) from
Achaetomium
sp. strain Xz8 via the optimization of charge-charge interactions. By using the enzyme thermal stability system (ETSS), two residues—D244 and D299—were inferred to be crucial contributors to thermostability. Single (D244A and D299R) and double (D244A/D299R) mutants were then generated and compared with the wild type. All mutants showed improved thermal properties, in the order D244A < D299R < D244A/D299R. In comparison with PG8fn, D244A/D299R showed the most pronounced shifts in temperature of maximum enzymatic activity (
T
max
), temperature at which 50% of the maximal activity of an enzyme is retained (
T
50
), and melting temperature (

T
m

), of about 10, 17, and 10.2°C upward, respectively, with the half-life (
t
1/2
) extended by 8.4 h at 50°C and 45 min at 55°C. Another distinguishing characteristic of the D244A/D299R mutant was its catalytic activity, which was comparable to that of the wild type (23,000 ± 130 U/mg versus 28,000 ± 293 U/mg); on the other hand, it showed more residual activity (8,400 ± 83 U/mg versus 1,400 ± 57 U/mg) after the feed pelleting process (80°C and 30 min). Molecular dynamics (MD) simulation studies indicated that mutations at sites D244 and D299 lowered the overall root mean square deviation (RMSD) and consequently increased the protein rigidity. This study reveals the importance of charge-charge interactions in protein conformation and provides a viable strategy for enhancing protein stability.
Exposure of Salmonella enterica Serovar Typhimurium to Three Humectants Used in the Food Industry Induces Different Osmoadaptation Systems
Authors: Finn Sarah , Rogers Lisa , Händler Kristian , McClure Peter , Amézquita Alejandro , Hinton Jay C. D. , Fanning Séamus
Published: July 2015

Common salt (NaCl) is frequently used by the food industry to add flavor and to act as a humectant in order to reduce the water content of a food product. The improved health awareness of consumers is leading to a demand for food products with reduced salt content; thus, manufacturers require alternative water activity-reducing agents which elicit the same general effects as NaCl. Two examples include KCl and glycerol. These agents lower the water activity of a food matrix and also contribute to limit the growth of the microbiota, including foodborne pathogens. Little is currently known about how foodborne pathogens respond to these water activity-lowering agents. Here we examined the response of
Salmonella enterica
serovar Typhimurium 4/74 to NaCl, KCl, and glycerol at three time points, using a constant water activity level, compared with the response of a control inoculum. All conditions induced the upregulation of gluconate metabolic genes after 6 h of exposure. Bacteria exposed to NaCl and KCl demonstrated the upregulation of the osmoprotective transporter mechanisms encoded by the
proP
,
proU
, and
osmU
(STM1491 to STM1494) genes. Glycerol exposure elicited the downregulation of these osmoadaptive mechanisms but stimulated an increase in lipopolysaccharide and membrane protein-associated genes after 1 h. The most extensive changes in gene expression occurred following exposure to KCl. Because many of these genes were of unknown function, further characterization may identify KCl-specific adaptive processes that are not stimulated by NaCl. This study shows that the response of
S
. Typhimurium to different humectants does not simply reflect reduced water activity and likely involves systems that are linked to specific humectants.
Novel Glucose-1-Phosphatase with High Phytase Activity and Unusual Metal Ion Activation from Soil Bacterium Pantoea sp. Strain 3.5.1
Authors: Suleimanova Aliya D. , Beinhauer Astrid , Valeeva Liia R. , Chastukhina Inna B. , Balaban Nelly P. , Shakirov Eugene V. , Greiner Ralf , Sharipova Margarita R.
Published: July 2015

Phosphorus is an important macronutrient, but its availability in soil is limited. Many soil microorganisms improve the bioavailability of phosphate by releasing it from various organic compounds, including phytate. To investigate the diversity of phytate-hydrolyzing bacteria in soil, we sampled soils of various ecological habitats, including forest, private homesteads, large agricultural complexes, and urban landscapes. Bacterial isolate
Pantoea
sp. strain 3.5.1 with the highest level of phytase activity was isolated from forest soil and investigated further. The
Pantoea
sp. 3.5.1
agpP
gene encoding a novel glucose-1-phosphatase with high phytase activity was identified, and the corresponding protein was purified to apparent homogeneity, sequenced by mass spectroscopy, and biochemically characterized. The AgpP enzyme exhibits maximum activity and stability at pH 4.5 and at 37°C. The enzyme belongs to a group of histidine acid phosphatases and has the lowest

K
m

values toward phytate, glucose-6-phosphate, and glucose-1-phosphate. Unexpectedly, stimulation of enzymatic activity by several divalent metal ions was observed for the AgpP enzyme. High-performance liquid chromatography (HPLC) and high-performance ion chromatography (HPIC) analyses of phytate hydrolysis products identify
dl
-
myo
-inositol 1,2,4,5,6-pentakisphosphate as the final product of the reaction, indicating that the
Pantoea
sp. AgpP glucose-1-phosphatase can be classified as a 3-phytase. The identification of the
Pantoea
sp. AgpP phytase and its unusual regulation by metal ions highlight the remarkable diversity of phosphorus metabolism regulation in soil bacteria. Furthermore, our data indicate that natural forest soils harbor rich reservoirs of novel phytate-hydrolyzing enzymes with unique biochemical features.
Origin of the Outbreak in France of Pseudomonas syringae pv. actinidiae Biovar 3, the Causal Agent of Bacterial Canker of Kiwifruit, Revealed by a Multilocus Variable-Number Tandem-Repeat Analysis
Authors: Cunty A. , Cesbron S. , Poliakoff F. , Jacques M.-A. , Manceau C.
Published: July 2015

The first outbreaks of bacterial canker of kiwifruit caused byPseudomonas syringaepv. actinidiae biovar 3 were detected in France in 2010.P. syringaepv. actinidiae causes leaf spots, dieback, and canker that sometimes lead to the death of the vine.P. syringaepv. actinidifoliorum, which is pathogenic on kiwi as well, causes only leaf spots. In order to conduct an epidemiological study to track the spread of the epidemics of these two pathogens in France, we developed a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA). MLVA was conducted on 340 strains ofP. syringaepv. actinidiae biovar 3 isolated in Chile, China, France, Italy, and New Zealand and on 39 strains ofP. syringaepv. actinidifoliorum isolated in Australia, France, and New Zealand. Eleven polymorphic VNTR loci were identified in the genomes ofP. syringaepv. actinidiae biovar 3 ICMP 18744 and ofP. syringaepv. actinidifoliorum ICMP 18807. MLVA enabled the structuring ofP. syringaepv. actinidiae biovar 3 andP. syringaepv. actinidifoliorum strains in 55 and 16 haplotypes, respectively. MLVA and discriminant analysis of principal components revealed that strains isolated in Chile, China, and New Zealand are genetically distinct fromP. syringaepv. actinidiae strains isolated in France and in Italy, which appear to be closely related at the genetic level. In contrast, no structuring was observed forP. syringaepv. actinidifoliorum. We developed an MLVA scheme to explore the diversity withinP. syringaepv. actinidiae biovar 3 and to trace the dispersal routes of epidemicP. syringaepv. actinidiae biovar 3 in Europe. We suggest using this MLVA scheme to trace the dispersal routes ofP. syringaepv. actinidiae at a global level.
The Bacterial Communities of Full-Scale Biologically Active, Granular Activated Carbon Filters Are Stable and Diverse and Potentially Contain Novel Ammonia-Oxidizing Microorganisms
Authors: LaPara Timothy M. , Hope Wilkinson Katheryn , Strait Jacqueline M. , Hozalski Raymond M. , Sadowksy Michael J. , Hamilton Matthew J.
Published: July 2015

The bacterial community composition of the full-scale biologically active, granular activated carbon (BAC) filters operated at the St. Paul Regional Water Services (SPRWS) was investigated using Illumina MiSeq analysis of PCR-amplified 16S rRNA gene fragments. These bacterial communities were consistently diverse (Shannon index, >4.4; richness estimates, >1,500 unique operational taxonomic units [OTUs]) throughout the duration of the 12-month study period. In addition, only modest shifts in the quantities of individual bacterial populations were observed; of the 15 most prominent OTUs, the most highly variable population (a
Variovorax
sp.) modulated less than 13-fold over time and less than 8-fold from filter to filter. The most prominent population in the profiles was a
Nitrospira
sp., representing 13 to 21% of the community. Interestingly, very few of the known ammonia-oxidizing bacteria (AOB; <0.07%) and no ammonia-oxidizing
Archaea
were detected in the profiles. Quantitative PCR of
amoA
genes, however, suggested that AOB were prominent in the bacterial communities (
amoA
/16S rRNA gene ratio, 1 to 10%). We conclude, therefore, that the BAC filters at the SPRWS potentially contained significant numbers of unidentified and novel ammonia-oxidizing microorganisms that possess
amoA
genes similar to those of previously described AOB.
Distinct SagA from Hospital-Associated Clade A1 Enterococcus faecium Strains Contributes to Biofilm Formation
Authors: Paganelli F. L. , de Been M. , Braat J. C. , Hoogenboezem T. , Vink C. , Bayjanov J. , Rogers M. R. C. , Huebner J. , Bonten M. J. M. , Willems R. J. L. , Leavis H. L.
Published: July 2015

Enterococcus faecium
is an important nosocomial pathogen causing biofilm-mediated infections. Elucidation of
E. faecium
biofilm pathogenesis is pivotal for the development of new strategies to treat these infections. In several bacteria, extracellular DNA (eDNA) and proteins act as matrix components contributing to biofilm development. In this study, we investigated biofilm formation capacity and the roles of eDNA and secreted proteins for 83
E. faecium
strains with different phylogenetic origins that clustered in clade A1 and clade B. Although there was no significant difference in biofilm formation between
E. faecium
strains from these two clades, the addition of DNase I or proteinase K to biofilms demonstrated that eDNA is essential for biofilm formation in most
E. faecium
strains, whereas proteolysis impacted primarily biofilms of
E. faecium
clade A1 strains. Secreted antigen A (SagA) was the most abundant protein in biofilms from
E. faecium
clade A1 and B strains, although its localization differed between the two groups.
sagA
was present in all sequenced
E. faecium
strains, with a consistent difference in the repeat region between the clades, which correlated with the susceptibility of biofilms to proteinase K. This indicates an association between the SagA variable repeat profile and the localization and contribution of SagA in
E. faecium
biofilms.
Transcriptomic Analysis of the Adaptation of Listeria monocytogenes to Growth on Vacuum-Packed Cold Smoked Salmon
Authors: Tang Silin , Orsi Renato H. , den Bakker Henk C. , Wiedmann Martin , Boor Kathryn J. , Bergholz Teresa M.
Published: July 2015

The foodborne pathogen
Listeria monocytogenes
is able to survive and grow in ready-to-eat foods, in which it is likely to experience a number of environmental stresses due to refrigerated storage and the physicochemical properties of the food. Little is known about the specific molecular mechanisms underlying survival and growth of
L. monocytogenes
under different complex conditions on/in specific food matrices. Transcriptome sequencing (RNA-seq) was used to understand the transcriptional landscape of
L. monocytogenes
strain H7858 grown on cold smoked salmon (CSS; water phase salt, 4.65%; pH 6.1) relative to that in modified brain heart infusion broth (MBHIB; water phase salt, 4.65%; pH 6.1) at 7°C. Significant differential transcription of 149 genes was observed (false-discovery rate [FDR], <0.05; fold change, ≥2.5), and 88 and 61 genes were up- and downregulated, respectively, in H7858 grown on CSS relative to the genes in H7858 grown in MBHIB. In spite of these differences in transcriptomes under these two conditions, growth parameters for
L. monocytogenes
were not significantly different between CSS and MBHIB, indicating that the transcriptomic differences reflect how
L. monocytogenes
is able to facilitate growth under these different conditions. Differential expression analysis and Gene Ontology enrichment analysis indicated that genes encoding proteins involved in cobalamin biosynthesis as well as ethanolamine and 1,2-propanediol utilization have significantly higher transcript levels in H7858 grown on CSS than in that grown in MBHIB. Our data identify specific transcriptional profiles of
L. monocytogenes
growing on vacuum-packaged CSS, which may provide targets for the development of novel and improved strategies to control
L. monocytogenes
growth on this ready-to-eat food.
Characterization of
para
-Nitrophenol-Degrading Bacterial Communities in River Water by Using Functional Markers and Stable Isotope Probing
Authors: Kowalczyk Agnieszka , Eyice Özge , Schäfer Hendrik , Price Oliver R. , Finnegan Christopher J. , van Egmond Roger A. , Shaw Liz J. , Barrett Glyn , Bending Gary D.
Published: July 2015

Microbial degradation is a major determinant of the fate of pollutants in the environment.
para
-Nitrophenol (PNP) is an EPA-listed priority pollutant with a wide environmental distribution, but little is known about the microorganisms that degrade it in the environment. We studied the diversity of active PNP-degrading bacterial populations in river water using a novel functional marker approach coupled with [
13
C
6
]PNP stable isotope probing (SIP). Culturing together with culture-independent terminal restriction fragment length polymorphism analysis of 16S rRNA gene amplicons identified
Pseudomonas syringae
to be the major driver of PNP degradation in river water microcosms. This was confirmed by SIP-pyrosequencing of amplified 16S rRNA. Similarly, functional gene analysis showed that degradation followed the Gram-negative bacterial pathway and involved
pnpA
from
Pseudomonas
spp. However, analysis of maleylacetate reductase (encoded by
mar
), an enzyme common to late stages of both Gram-negative and Gram-positive bacterial PNP degradation pathways, identified a diverse assemblage of bacteria associated with PNP degradation, suggesting that
mar
has limited use as a specific marker of PNP biodegradation. Both the
pnpA
and
mar
genes were detected in a PNP-degrading isolate,
P. syringae
AKHD2, which was isolated from river water. Our results suggest that PNP-degrading cultures of
Pseudomonas
spp. are representative of environmental PNP-degrading populations.
Topographical Mapping of the Rainbow Trout (Oncorhynchus mykiss) Microbiome Reveals a Diverse Bacterial Community with Antifungal Properties in the Skin
Authors: Lowrey Liam , Woodhams Douglas C. , Tacchi Luca , Salinas Irene
Published: July 2015

The mucosal surfaces of wild and farmed aquatic vertebrates face the threat of many aquatic pathogens, including fungi. These surfaces are colonized by diverse symbiotic bacterial communities that may contribute to fight infection. Whereas the gut microbiome of teleosts has been extensively studied using pyrosequencing, this tool has rarely been employed to study the compositions of the bacterial communities present on other teleost mucosal surfaces. Here we provide a topographical map of the mucosal microbiome of an aquatic vertebrate, the rainbow trout (
Oncorhynchus mykiss
). Using 16S rRNA pyrosequencing, we revealed novel bacterial diversity at each of the five body sites sampled and showed that body site is a strong predictor of community composition. The skin exhibited the highest diversity, followed by the olfactory organ, gills, and gut.
Flectobacillus
was highly represented within skin and gill communities. Principal coordinate analysis and plots revealed clustering of external sites apart from internal sites. A highly diverse community was present within the epithelium, as demonstrated by confocal microscopy and pyrosequencing. Using
in vitro
assays, we demonstrated that two
Arthrobacter
sp. skin isolates, a
Psychrobacter
sp. strain, and a combined skin aerobic bacterial sample inhibit the growth of
Saprolegnia australis
and
Mucor hiemalis
, two important aquatic fungal pathogens. These results underscore the importance of symbiotic bacterial communities of fish and their potential role for the control of aquatic fungal diseases.
Effects of Chicken Litter Storage Time and Ammonia Content on Thermal Resistance of Desiccation-Adapted Salmonella spp
Authors: Chen Zhao , Wang Hongye , Ionita Claudia , Luo Feng , Jiang Xiuping
Published: July 2015

Broiler chicken litter was kept as a stacked heap on a poultry farm, and samples were collected up to 9 months of storage. Chicken litter inoculated with desiccation-adapted
Salmonella
cells was heat-treated at 75, 80, 85, and 150°C.
Salmonella
populations decreased in all these samples during heat treatment, and the inactivation rates became lower in chicken litter when storage time was extended from 0 to 6 months. There was no significant difference (
P
> 0.05) in thermal resistance of
Salmonella
in 6- and 9-month litter samples, indicating that a threshold for thermal resistance was reached after 6 months. Overall, the thermal resistance of
Salmonella
in chicken litter was affected by the storage time of the litter. The changes in some chemical, physical, and microbiological properties during storage could possibly contribute to this difference. Moisture and ammonia could be two of the most significant factors influencing the thermal resistance of
Salmonella
cells in chicken litter. Our results emphasize the importance of adjusting time and temperature conditions for heat processing chicken litter when it is removed from the chicken house at different time intervals.
Aspergillus glaucus Aquaglyceroporin Gene
glpF
Confers High Osmosis Tolerance in Heterologous Organisms
Authors: Liu Xiao-Dan , Wei Yi , Zhou Xiao-Yang , Pei Xue , Zhang Shi-Hong
Published: July 2015

Aquaglyceroporins (GlpFs) that transport glycerol along with water and other uncharged solutes are involved in osmoregulation in myriad species. Fungal species form a large group of eukaryotic organisms, and their GlpFs may be diverse, exhibiting various activities. However, few filamentous fungal GlpFs have been biologically investigated. Here, a
glpF
gene from the halophilic fungus
Aspergillus glaucus
(
AgglpF
) was verified to be a channel of water or glycerol in
Xenopus laevis
oocytes and was further functionally analyzed in three heterologous systems. In
Saccharomyces cerevisiae
, cells overexpressing
AgglpF
possessed significant tolerance of drought, salt, and certain metal ions.
AgglpF
was then characterized in the filamentous fungus of
Neurospora crassa
. Based on the
N. crassa
aquaporin gene (
NcAQP
) disruption mutant (the Δ
aqp
mutant), a series of complementary strains carrying
NcAQP
and
AgglpF
and three asparagine-proline-alanine-gene (
NPA
)-deleted
AgglpF
fragments were created. As revealed by salt resistance analysis, the
AgglpF
complementary strain possessed the highest salt resistance among the tested strains. In addition, the intracellular glycerol content in the
AgglpF
complementary strain was markedly higher than that in the other strains. The AgGlpF-green fluorescent protein (GFP) fusion protein was subcellularly localized in the plasma membrane of onion epidermal cells, suggesting that
AgglpF
functions in plants. Indeed, when
AgglpF
was expressed in
Arabidopsis thaliana
, transgenic lines survived under conditions of high osmotic stress and under conditions of drought stress in particular. Overall, our results revealed that AgGlpF as a water/glycerol transporter is required for survival of both fungi and plants under conditions of high osmotic stress and may have value in applications in genetic engineering for generating high salt and drought resistance.
The Octatricopeptide Repeat Protein Raa8 Is Required for Chloroplast
trans
Splicing
Authors: Marx Christina , Wünsch Christiane , Kück Ulrich
Published: July 2015

The mRNA maturation of the tripartite chloroplast
psaA
gene from the green alga
Chlamydomonas reinhardtii
depends on various nucleus-encoded factors that participate in
trans
splicing of two group II introns. Recently, a multiprotein complex was identified that is involved in processing the
psaA
precursor mRNA. Using coupled tandem affinity purification (TAP) and mass spectrometry analyses with the
trans
-splicing factor Raa4 as a bait protein, we recently identified a multisubunit ribonucleoprotein (RNP) complex comprising the previously characterized
trans
-splicing factors Raa1, Raa3, Raa4, and Rat2 plus novel components. Raa1 and Rat2 share a structural motif, an octatricopeptide repeat (OPR), that presumably functions as an RNA interaction module. Two of the novel RNP complex components also exhibit a predicted OPR motif and were therefore considered potential
trans
-splicing factors. In this study, we selected bacterial artificial chromosome (BAC) clones encoding these OPR proteins and conducted functional complementation assays using previously generated
trans
-splicing mutants. Our assay revealed that the
trans
-splicing defect of mutant F19 was restored by a new factor we named
RAA8
; molecular characterization of complemented strains verified that Raa8 participates in splicing of the first
psaA
group II intron. Three of six OPR motifs are located in the C-terminal end of Raa8, which was shown to be essential for restoring
psaA
mRNA
trans
splicing. Our results support the important role played by OPR proteins in chloroplast RNA metabolism and also demonstrate that combining TAP and mass spectrometry with functional complementation studies represents a vigorous tool for identifying
trans
-splicing factors.
Management of developmental speech and language disorders: Part 1
Authors: O'Hare Anne , Bremner Lynne
Published: July 2015

The identification of developmental problems in a child's acquisition of speech, language and/or communication is a core activity in child surveillance. These are common difficulties with up to 15% of toddlers being ‘late talkers’ and 7% of children entering school with persisting impairments of their language development. These delays can confer disadvantages in the long term, adversely affecting language, cognition, academic attainment, behaviour and mental health. All children presenting with significant speech and language delay should be investigated with a comprehensive hearing assessment and be considered for speech and language therapy assessment. Socioeconomic adversity correlates with delayed language development. Clinical assessment should confirm that the presentation is definitely not acquired (see part 2) and will also guide whether the difficulty is primary, in which there are often familial patterns, or secondary, from a very wide range of aetiologies. Symptoms may be salient, such as the regression of communication in <3-year-olds which ‘flags up’ autism spectrum disorder. Further investigation will be informed from this clinical assessment, for example, genetic investigation for sex aneuploidies in enduring primary difficulties. Management of the speech and language difficulty itself is the realm of the speech and language therapist, who has an ever-increasing evidence-based choice of interventions. This should take place within a multidisciplinary team, particularly for children with more severe conditions who may benefit from individualised parental and educational supports.
Variation in cervical and breast cancer screening coverage in England: a cross-sectional analysis to characterise districts with atypical behaviour
Authors: Massat Nathalie J , Douglas Elaine , Waller Jo , Wardle Jane , Duffy Stephen W
Published: July 2015

Objectives

Reducing cancer screening inequalities in England is a major focus of the 2011 Department of Health cancer outcome strategy. Screening coverage requires regular monitoring in order to implement targeted interventions where coverage is low. This study aimed to characterise districts with atypical coverage levels for cervical or breast screening.

Design

Observational study of district-level coverage in the English Cervical and Breast screening programmes in 2012.

Setting

England, UK.

Participants

All English women invited to participate in the cervical (age group 25–49 and 50–64) and breast (age group 50–64) screening programmes.

Outcomes

Risk adjustment models for coverage were developed based on district-level characteristics. Funnel plots of adjusted coverage were constructed, and atypical districts examined by correlation analysis.

Results

Variability in coverage was primarily explained by population factors, whereas general practice characteristics had little independent effect. Deprivation and ethnicity other than white, Asian, black or mixed were independently associated with poorer coverage in both screening programmes, with ethnicity having the strongest effect; by comparison, the influence of Asian, black or mixed ethnic minority was limited. Deprivation, ethnicity and urbanisation largely accounted for the lower cervical screening coverage in London. However, for breast screening, being located in London remained a strong negative predictor. A subset of districts was identified as having atypical coverage across programmes. Correlates of deprivation in districts with relatively low adjusted coverage were substantially different from overall correlates of deprivation.

Discussion

These results inform the continuing drive to reduce avoidable cancer deaths in England, and encourage implementation of targeted interventions in communities residing in districts identified as having atypically low coverage. Sequential implementation to monitor the impact of local interventions would help accrue evidence on ‘what works’.
A questionnaire survey exploring healthcare professionals’ attitudes towards teamwork and safety in acute care areas in South Korea
Authors: Kim Sung Eun , Kim Chan Woong , Lee Sang Jin , Oh Je Hyeok , Lee Dong Hoon , Lim Tae Ho , Choi Hyuk Joong , Chung Hyun Soo , Ryu Ji Yeong , Jang Hye Young , Choi Yoon Hee , Kim Su Jin , Jung Jin Hee
Published: July 2015

Objectives

Although human factors are important in terms of patient safety, there have been very few reports on the attitudes of healthcare professionals working in acute care settings in South Korea. In the present study, we investigated the attitudes of such professionals, their cultures and their management systems.

Design

A questionnaire survey with 65 items covering nine themes affecting patient safety. Nine themes were compared via a three-or-more-way analysis of variance, with interaction, followed by multiple comparisons among several groups.

Setting

Intensive care units, emergency departments and surgical units of nine urban hospitals.

Participants

592 nurses and 160 physicians.

Intervention

None.

Outcome measures

Mean scores using a five-point scale and combined response scores for each of the nine themes.

Results

The mean score for information-sharing was the highest (3.78±0.49) and that for confidence/assertion was the lowest (2.97±0.34). The mean scores for teamwork, error management, work value, organisational climate, leadership, stress and fatigue level, and error/procedural compliance were intermediate. Physicians showed lower scores in leadership and higher scores in information-sharing than nurses. Respondents with 24 months or less of a clinical career showed higher scores in leadership, stress and fatigue, and error scores and lower scores in work value than more experienced respondents.

Conclusions

Our results suggest that medical personnel in Korea are relatively reluctant to disclose error or assert their different opinions with others. Many did not adequately recognise the negative effects of fatigue and stress, attributed errors to personal incompetence, and error-management systems were inadequate. Discrepancies in leadership and information-sharing were evident between professional groups, and leadership, stress, fatigue level, work value and error scores varied with the length of work experience. These can be used as baseline data to establish training programmes for patient safety in Korea.
‘Seizure First Aid Training’ for people with epilepsy who attend emergency departments, and their family and friends: study protocol for intervention development and a pilot randomised controlled trial
Authors: Noble AJ , Marson AG , Tudur-Smith C , Morgan M , Hughes DA , Goodacre S , Ridsdale L
Published: July 2015

Introduction

People with chronic epilepsy (PWE) often make costly but clinically unnecessary emergency department (ED) visits. Offering them and their carers a self-management intervention that improves confidence and ability to manage seizures may lead to fewer visits. As no such intervention currently exists, we describe a project to develop and pilot one.

Methods and analysis

To develop the intervention, an existing group-based seizure management course that has been offered by the Epilepsy Society within the voluntary sector to a broader audience will be adapted. Feedback from PWE, carers and representatives from the main groups caring for PWE will help refine the course so that it addresses the needs of ED attendees. Its behaviour change potential will also be optimised. A pilot randomised controlled trial will then be completed. 80 PWE aged ≥16 who have visited the ED in the prior 12 months on ≥2 occasions, along with one of their family members or friends, will be recruited from three NHS EDs. Dyads will be randomised to receive the intervention or treatment as usual alone. The proposed primary outcome is ED use in the 12 months following randomisation. For the pilot, this will be measured using routine hospital data. Secondary outcomes will be measured by patients and carers completing questionnaires 3, 6 and 12 months postrandomisation. Rates of recruitment, retention and unblinding will be calculated, along with the ED event rate in the control group and an estimate of the intervention's effect on the outcome measures.

Ethics and dissemination

Ethical approval: NRES Committee North West—Liverpool East (Reference number 15/NW/0225). The project's findings will provide robust evidence on the acceptability of seizure management training and on the optimal design of a future definitive trial. The findings will be published in peer-reviewed journals and presented at conferences.

Trial registration number

ISRCTN13 871 327.
Safety culture in long-term care: a cross-sectional analysis of the Safety Attitudes Questionnaire in nursing and residential homes in the Netherlands
Authors: Buljac-Samardzic Martina , van Wijngaarden Jeroen DH , Dekker–van Doorn Connie M
Published: July 2015

Objective

The first objective was to investigate if the Safety Attitudes Questionnaire (SAQ) is appropriate to measure the safety attitude of caregivers in nursing and residential homes, and second, to compare safety attitude of these caregivers with available data of caregivers in other settings (ie, inpatients, intensive care unit (ICU) and ambulatory care).

Methods

Using a cross-sectional survey methodology, we obtained completed SAQ surveys from 521 caregivers (response rate of 53%) working in nine units in nine different nursing and residential homes in The Netherlands. Exploratory factor and Cronbach's alpha measures were used to analyse the psychometric properties of the SAQ. A correlation matrix was performed to study the relationship among the SAQ dimensions. A t test was performed to test significant differences between our sample and the benchmark settings.

Results

The factor analyses and calculated Cronbach's alphas (α=0.56–0.80) for this sample confirmed the robustness of the SAQ scales. There was a high positive correlation between teamwork climate, job satisfaction, perceptions of management, safety climate and working conditions (r=0.31 to 63), but stress recognition had a negative correlation with each of the other dimensions (r=−0.13 to −0.18). Overall, the scores from the nursing and residential homes differed significantly from the benchmark settings.

Conclusions

The findings in this study confirmed that the SAQ could also be used in the nursing and residential homes setting. However, stress recognition in nursing and residential homes setting does not seem to be one of the dimensions of the safety attitude construct. Furthermore, Dutch nursing and residential homes have significantly higher scores on most dimensions of the SAQ compared with US inpatient units and comparable scores to ICUs (Dutch and US) and ambulatory services.
What do patients say about emergency departments in online reviews? A qualitative study
Authors: Kilaru Austin S , Meisel Zachary F , Paciotti Breah , Ha Yoonhee P , Smith Robert J , Ranard Benjamin L , Merchant Raina M
Published: July 2015

Background

Patients have adopted web-based tools to report on the quality of their healthcare experiences. We seek to examine online reviews for US emergency departments (EDs) posted on Yelp, a popular consumer ratings website.

Methods

We conducted a qualitative analysis of unstructured, publicly accessible reviews for hospitals available onhttp://www.yelp.com. We collected all reviews describing experiences of ED care for a stratified random sample of 100 US hospitals. We analysed the content of the reviews using themes derived from the Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) inpatient care survey. We also used modified grounded theory to iteratively code the text of the reviews, identifying additional themes specific to emergency care. The data were double-coded, and discrepancies were evaluated to ensure consensus.

Results

Of the 1736 total reviews, 573 (33%) described patient experiences involving the ED. The reviews contained several themes assessed by the HCAHPS survey, including communication with nurses, communication with doctors, and pain control. The reviews also contained key themes specific to emergency care: waiting and efficiency; decisions to seek care in the ED; and events following discharge, including administrative difficulties.

Conclusions

These exploratory findings suggest that online reviews for EDs contain similar themes to survey-based assessments of inpatient hospital care as well as themes specific to emergency care. Consumer rating websites allow patients to provide rapid and public feedback on their experience of medical care. Web-based platforms may offer a novel strategy for assessing patient-centred quality in emergency care.
Implementation of HIV treatment as prevention strategy in 17 Canadian sites: immediate and sustained outcomes from a 35-month Quality Improvement Collaborative
Authors: Clarke Christina M , Cheng Tessa , Reims Kathleen G , Steinbock Clemens M , Thumath Meaghan , Milligan Robert Sam , Barrios Rolando
Published: July 2015

Background

Rapid scale-up of effective antiretroviral therapy (ART) is required to meet global targets to eliminate new HIV infections and AIDS-related deaths. Yet, gaps persist in all nations striving for these targets. In the intervention setting of British Columbia (BC), Canada, where ART is publicly funded, 73% of HIV-diagnosed were on ART in 2011, and only 49% were achieving viral suppression.

Methods

An observational case descriptive study of HIV care sites in BC recruited to participate in a 35-month Breakthrough Series Quality Improvement Collaborative and sustainability network. Sites collected four quality indicators, qualitative change descriptions and implemented the chronic care model (CCM) and HIV care and treatment guidelines. Two reviewers assigned monthly implementation scores to evaluate site progress (January 2011–2012). All quality indicators were pooled and analysed using probability-based run chart rules.

Results

Seventeen teams with a pooled median population of 2296 HIV patients joined the initiative. Comprehensive CCM implementation and evidence of improvement was achieved by 29% of sites (implementation score of 4.0 or higher on 5.0 scale). Evidence of sustained improvement was observed for patient engagement (88.8–90.4%), ART uptake among patients unequivocally in need (92.9–94.8%), and ART uptake (≥6 months) and achieving viral suppression (57.3–78.4%) (all p<0.05).

Conclusions

This study shows evidence of sustained improvements in HIV care processes and treatment outcomes for an estimated population of 2296 HIV patients in 17 BC sites. Overall success points to opportunities for other high-income countries seeking to improve HIV health outcomes.
Changing an ingrained culture: Improving the safety of oxygen therapy at University Hospitals Bristol NHS Foundation Trust
Authors: Gatter Michelle , Dixon Giles , Wall Josh , Mew Ed
Published: July 2015

Oxygen is one of the most commonly administered drugs in UK hospitals. Our quality improvement project aimed to increase the safety of oxygen therapy at University Hospitals Bristol NHS Foundation Trust. We aimed to increase the rate of oxygen prescribing and increase the percentage of nurses signing appropriately for oxygen titration and administration. We hypothesised this would result in a higher percentage of patients achieving their appropriate oxygen saturations.

Our project ran on several acute medical and surgical wards. We tested several interventions with a plan, do, study, act method of continuous data collection. We firstly focussed on the education of junior doctors and then the wider multi-disciplinary team with a trust-wide “safety focus”. We utilised patient safety systems already in place in the hospital, such as the clinical risk register and incident reporting system. We also trialled an intervention that was successfully implemented by another group in a different trust in the UK.

Oxygen prescription increased from 44.4% to 76.9% over the duration of the project. Appropriate nursing signatures increased from 26.6% to 60%. The number of patients achieving appropriate target saturations rose from 61.8% to 76.7%. The most successful interventions were the trust safety briefing and oxygen safety hangers.

Our project has showed the importance of integrating new projects within safety schemes already available. Persistence and careful intervention are key to changing strongly engrained cultures in large organisations. Interventions that have proved to be successful in other trusts can be implemented to enact change.
Association between patterns of leisure time physical activity and asthma control in adult patients
Authors: Bacon Simon L , Lemiere Catherine , Moullec Gregory , Ninot Gregory , Pepin Véronique , Lavoie Kim L
Published: July 2015

Background

Physical activity has been shown to have various health benefits in patients with asthma, especially in children. However, there are still limited data on the nature of the association between physical activity and asthma control in adults.

Objective

The objective of the current study was to determine the nature of the association between physical activity and asthma control, with particular emphasis on the intensity of the activity and seasonal variations.

Methods

643 adult patients with objectively confirmed asthma (mean age (SD)=53 (15) years, 60% women) were interviewed by telephone. Patients completed the asthma control questionnaire (ACQ), the asthma quality of life questionnaire, and a 1-year physical activity recall questionnaire to assess leisure time physical activity (LTPA).

Results

Total LTPA was related to control (β (95% CI)=−0.013 (−0.030 to 0.006)), with those doing recommended levels of LTPA being nearly 2.5 times more likely to have good control compared with inactive patients. Analysis of seasonal exercise habits found that winter LTPA (β=−0.027 (−0.048 to −0.006)) was more strongly associated with ACQ scores than summer LTPA (β=−0.019 (−0.037 to −0.001)). Adjustment for age, sex, season of assessment, inhaled corticosteroid (ICS) dose, body mass index, and current smoking status reduced the strength of the relationships.

Conclusions

Data indicate that higher levels of LTPA are associated with better levels of asthma control in adult patients with asthma, and that this seems to be more pronounced among asthmatics who do the recommended levels of exercise.
Palliative care for cirrhosis: a UK survey of health professionals’ perceptions, current practice and future needs
Authors: Low Joseph , Vickerstaff Victoria , Davis Sarah , Bichard Julia , Greenslade Lynda , Hopkins Katherine , Marshall Aileen , Thorburn Douglas , Jones Louise
Published: July 2015

Objective

To determine the knowledge and practice patterns of a UK cohort of relevant healthcare professionals (HCPs) about delivering palliative care in cirrhosis, and to inform priorities for future research.

Design

An on-line questionnaire survey with closed and open responses.

Setting

HCPs identified from the mailing list of special interest groups in hepatology and gastroenterology (liver), general practice and specialist palliative care (SPC) across the UK.

Results

Of the 6181 potential contacts identified, 517 HCPs responded. Most believed a role exists for SPC in caring for people with cirrhosis, but many SPC HCPs felt ill prepared to provide good care to those facing death. Further training was needed in managing liver-related symptoms, symptom control and end of life issues. All HCP groups wished to increase community provision of palliative care support, but many general practitioners felt unable to manage advanced cirrhosis in the community. There were differences in the optimal trigger for SPC referral with liver HCPs less likely to refer at symptom deterioration. Prognostication, symptom management and service configuration were key areas identified for future research.

Conclusions

All who responded acknowledged the role of SPC in caring for those dying with cirrhosis and need for further training to improve confidence and enable joint working between SPC, general practice and liver teams. Low response rates make it difficult to generalise these findings, which require further validation.
Second harmonic generation microscopy is a novel technique for differential diagnosis of breast fibroepithelial lesions
Authors: Tan Wai Jin , Yan Jie , Xu Shuoyu , Thike Aye Aye , Bay Boon Huat , Yu Hanry , Tan Min-Han , Tan Puay Hoon
Published: July 2015

Breast fibroepithelial lesions, including fibroadenomas and phyllodes tumours, are commonly encountered in clinical practice. As histological differences between these two related entities may be subtle, resulting in a challenging differential diagnosis, pathological techniques to assist the differential diagnosis of these two entities are of high interest. An accurate diagnosis at biopsy is important given corresponding implications for clinical decision-making including surgical extent and monitoring. Second harmonic generation (SHG) microscopy is a recently developed optical imaging technique capable of robust, powerful and unbiased label-free direct detection of collagen fibril structure in tissue without the use of antibodies. We constructed tissue microarrays emulating limited materials on biopsy to investigate quantitative collagen signal in fibroepithelial lesions using SHG microscopy. Archived formalin-fixed paraffin-embedded materials of 47 fibroepithelial lesions (14 fibroadenomas and 33 phyllodes tumours) were evaluated. Higher collagen signal on SHG microscopy was observed in fibroadenomas than phyllodes tumours on SHG imaging (p<0.001, area under the curve 0.859). At an automated threshold (2.5 million positive pixels), the sensitivity and specificity of the SHG microscopy for fibroadenoma classification was 71.4% and 84.4%, respectively. To corroborate these findings, we performed immunohistochemistry on tissue array sections using collagen I and III primary antibodies. Both collagen I and III immunohistochemical expressions were also significantly higher in fibroadenomas than in phyllodes tumours (p<0.001). In conclusion, label-free collagen quantitation on SHG microscopy is a novel imaging approach that can aid the differential diagnosis of fibroepithelial lesions.
Detection of aneurysmal subarachnoid hemorrhage 3 months after initial bleeding: evaluation of T2* and FLAIR MR sequences at 3 T in comparison with initial non-enhanced CT as a gold standard
Authors: Mulé Sébastien , Soize Sébastien , Benaissa Azzedine , Portefaix Christophe , Pierot Laurent
Published: July 2015

Objective

To investigate the ability of T2* and fluid-attenuated inversion recovery (FLAIR) MR sequences to detect hemosiderin deposition 3 months after aneurysmal subarachnoid hemorrhage (SAH) in comparison with early non-enhanced CT (NECT) as a gold standard.

Materials and methods

From September 2008 through May 2013, patients with aneurysmal SAH were included if a NECT less than 24 h after the onset of symptoms showed a SAH, and MRI, including T2* and FLAIR sequences, was performed 3 months later. All aneurysms were treated endovascularly. NECT and MR sequences were blindly analyzed for the presence of SAH (NECT) or hemosiderin deposition (MRI). When positive, details of the spatial distribution of SAH or hemosiderin deposits were noted. Sensitivities were calculated for each patient. Sensitivities, specificities, and positive predictive values (PPVs) were calculated for each location.

Results

Forty-nine patients (mean age 52.9 years) were included. Bleeding-related patterns were identified in 43 patients (87.8%) on T2* and 10 patients (20.4%) on FLAIR. T2* was highly predictive of the location of the initial hemorrhage, especially in the Sylvian cisterns (PPVs 95% and 100%) and the anterior interhemispheric fissure (PPV 90%).

Conclusions

The T2* sequence can detect and localize a previous SAH a few months after aneurysmal bleeding.
Canine adipose‐derived stromal cell viability following exposure to synovial fluid from osteoarthritic joints
Authors: Kiefer Kristina M. , O'Brien Timothy D. , Pluhar Elizabeth G. , Conzemius Michael
Published: July 2015

Introduction

Stem cell therapy used in clinical application of osteoarthritis in veterinary medicine typically involves intra‐articular injection of the cells, however the effect of an osteoarthritic environment on the fate of the cells has not been investigated.

Aims and Objectives

Assess the viability of adipose derived stromal cells following exposure to osteoarthritic joint fluid.

Materials and Methods

Adipose derived stromal cells (ASCs) were derived from falciform adipose tissue of five adult dogs, and osteoarthritic synovial fluid (SF) was obtained from ten patients undergoing surgical intervention on orthopedic diseases with secondary osteoarthritis. Normal synovial fluid was obtained from seven adult dogs from an unrelated study. ASCs were exposed to the following treatment conditions: culture medium, normal SF, osteoarthritic SF, or serial dilutions of 1:1 to 1:10 of osteoarthritic SF with media. Cells were then harvested and assessed for viability using trypan blue dye exclusion.

Results

There was no significant difference in the viability of cells in culture medium or normal SF. Significant differences were found between cells exposed to any concentration of osteoarthritic SF and normal SF and between cells exposed to undiluted osteoarthritic SF and all serial dilutions. Subsequent dilutions reduced cytotoxicity.

Conclusions

Osteoarthritic synovial fluid in this ex vivo experiment is cytotoxic to ASCs, when compared with normal synovial fluid. Current practice of direct injection of ASCs into osteoarthritic joints should be re‐evaluated to determine if alternative means of administration may be more effective.