bluuebunny/crossref_metadata_2025 huggingface dataset:
Parents’ knowledge, attitudes, use of pain relief methods and satisfaction related to their children's postoperative pain management: a descriptive correlational study
Authors: Chng Hui Yi , He Hong‐Gu , Chan Sally Wai‐Chi , Liam Joanne Li Wee , Zhu Lixia , Cheng Karis Kin Fong
Published: February 2015

Aims and objectives

To examine parents’ knowledge about and attitudes towards pain management, use of pain relief strategies and satisfaction with their children's postoperative pain management, as well as the relationships among these variables.

Background

Children's postoperative pain is inadequately managed worldwide. Despite increasing emphasis on parental involvement in children's postoperative pain management, few studies have examined parents’ management of their child's postoperative pain in Singapore.

Design

A descriptive correlational study.

Methods

A convenience sample of 60 parents having 6‐ to 14‐year‐old children undergoing inpatient elective surgery in a public hospital in Singapore was recruited. Data were collected one day after each child's surgery using the Pain Management Knowledge and Attitudes questionnaire and the Parents’ Use of Pain Relief Strategies questionnaire. Descriptive statistics, Mann–Whitney U tests, Kruskal–Wallis tests and Spearman's rank correlation coefficient were used for data analyses.

Results

Parents displayed moderate levels of knowledge, attitudes and use of pain relief methods in relation to their children's postoperative pain and pain management. They were generally satisfied with the management of their child's postoperative pain. There was significant difference in Parents’ Use of Pain Relief Strategies scores between parents with and without previous experience of caring for their hospitalised child. Statistically significant positive correlations were found between parents’ knowledge and attitude, knowledge and satisfaction, attitude and satisfaction, knowledge and pain relief strategies, as well as attitude and pain relief strategies.

Conclusion

This study indicates the need to provide parents with more information regarding their children's postoperative pain management. Future studies are needed to develop educational interventions for parents to improve their knowledge and attitudes, which will eventually improve their use of pain relief strategies for their children.

Relevance to clinical practice

Health professionals need to provide more information to equip parents with knowledge and skills regarding paediatric postoperative pain management.
Increased risk of breakthrough infection among cytomegalovirus donor‐positive/recipient‐negative kidney transplant recipients receiving lower‐dose valganciclovir prophylaxis
Authors: Stevens D.R. , Sawinski D. , Blumberg E. , Galanakis N. , Bloom R.D. , Trofe‐Clark J.
Published: February 2015

Objective

We compared the effectiveness of lower‐dose (LD) (450 mg/day for 6 months) to standard‐dose (SD) (900 mg/day for 6 months) valganciclovir (VGCV) prophylaxis for prevention of cytomegalovirus (CMV) infection and disease in high‐risk CMV donor‐positive/recipient‐negative (D+/R−) kidney recipients.

Methods

We performed a single‐center, retrospective cohort study, in a 750‐bed academic medical center, involving a total of 90 evaluable CMV high‐risk kidney recipients. All patients were retrospectively followed from day of transplantation to November 1, 2012, or to the development of CMV infection or disease, death, or loss to follow‐up. CMV screening was only done if suggestive symptoms or abnormal laboratory values were present. Our immunosuppressive protocol otherwise did not differ between periods.

Results

In total, 45 consecutive eligible patients initiated SD prophylaxis in the 22 months before the institutional protocol change regarding CMV prophylaxis. One patient developed CMV infection in the setting of non‐adherence. In the 16 months after the protocol update, 45 consecutive eligible patients receiving LD prophylaxis were evaluated: 6 developed CMV infection while receiving prophylaxis (P = 0.11). Ganciclovir (GCV)‐resistant infection was confirmed in 1 patient in the LD prophylaxis group. Late‐onset CMV infection or disease occurred in 11 patients (24%) in the SD group and in 12 patients (27%) in the LD group (P = 0.86). More patients in the SD group developed leukopenia (75% vs. 44%, P < 0.01). During the study period, no significant differences were seen between the groups in mean mg/kg exposure to rabbit anti‐thymocyte globulin induction courses, mean tacrolimus troughs, number of rejection episodes, mean estimated renal function, graft survival, or patient survival. Overall mean follow‐up (± standard deviation) was 357 days (± 53) in the SD group and 320 days (± 103) in the LD group (P = 0.03).

Conclusion

Breakthrough CMV infection while receiving VGCV prophylaxis occurred more often after the institutional protocol revision to LD VGCV prophylaxis. Given our concern for increased risk of breakthrough infection and GCV resistance when prophylaxis is under‐dosed, our institutional protocols were revised back to SD prophylaxis for all CMV D+/R− kidney transplant recipients.
Expression activation and functional analysis of HLA3, a putative inorganic carbon transporter in Chlamydomonas reinhardtii
Authors: Gao Han , Wang Yingjun , Fei Xiaowen , Wright David A. , Spalding Martin H.
Published: February 2015

The CO2 concentrating mechanism (CCM) is a key component of the carbon assimilation strategy of aquatic microalgae. Induced by limiting CO2 and tightly regulated, the CCM enables these microalgae to respond rapidly to varying environmental CO2 supplies and to perform photosynthetic CO2 assimilation in a cost‐effective way. A functional CCM in eukaryotic algae requires Rubisco sequestration, rapid interconversion between CO2 and catalyzed by carbonic anhydrases (CAs), and active inorganic carbon (Ci) uptake. In the model microalga Chlamydomonas reinhardtii, a membrane protein HLA3 is proposed to be involved in active Ci uptake across the plasma membrane. In this study, we use an artificially designed transcription activator‐like effector (dTALE) to activate the expression of HLA3. The successful activation of HLA3 expression demonstrates dTALE as a promising tool for gene‐specific activation and investigation of gene function in Chlamydomonas. Activation of HLA3 expression in high CO2 acclimated cells, where HLA3 is not expressed, resulted in increased Ci accumulation and Ci‐dependent photosynthetic O2 evolution specifically in very low CO2 concentrations, which confirms that HLA3 is indeed involved in Ci uptake, and suggests it is mainly associated with transport in very low CO2 concentrations, conditions in which active CO2 uptake is highly limited.
Two-Dimensional Free Convection Heat Transfer Below a Horizontal Hot Isothermal Flat Strip
Authors: Samie Milad , Nouri Gheimassi Arash , Salari Alinaghi , Behshad Shafii Mohammad
Published: February 2015

Convection heat transfer below a horizontal, hot, and isothermal strip of infinite length and width of 2L embedded in fluids with different Prandtl number (Pr) and Nusselt number (Nu) is analyzed with the aid of integral method. A new concept is utilized to determine the boundary layer thickness at the strip's edge, which is based on matching the flow rate of the boundary layer below the strip at its edge and the flow rate of the plume, which forms after the heated fluid detaches from the strip's edge. In addition to these novelties, a numerical model is developed to verify the analytical framework, and an excellent agreement is observed between the analytical and numerical models.
Surface analysis of zinc-porphyrin functionalized carbon nano-onions
Authors: Spampinato Valentina , Ceccone Giacomo , Giordani Silvia
Published: February 2015

Multishell fullerenes, known as carbon nano-onions (CNOs), are an interesting class of carbon-based nanomaterials. They display several unique properties, such as a large surface area to volume ratio, a low density, and a graphitic multilayer morphology, which have made them appealing for several applications in many fields, including biology. Chemical functionalization of CNOs dramatically enhances their solubility and attenuates their inflammatory properties, thereby increasing their applicability especially in the fields of biology and medicine. CNOs functionalized with fluorescent probes can be used for cellular imaging. In this article, detailed surface characterization of CNOs functionalized with a zinc porphyrin (ZnTPP) as the fluorescent probe is presented. In particular, time-of-flight secondary ion mass spectrometry and x-ray photoelectron spectroscopy provide a detailed surface characterization of the organic functionalities introduced via “click chemistry” and clearly demonstrate the success of the CNOs functionalization process. XPS data reveal the presence of Zn and N, whilst ToF-SIMS is able to identify specific fragments related to the presence of the ZnTPP, such as the quasimolecular mass peak [C32N4H20Zn]+ and the molecular mass peak [C44N4H28Zn]+. Moreover, ions fragments deriving from the triazole ring formed by the click chemistry reaction have been identified by ToF-SIMS analysis, proving unambiguously the covalent binding of the fluorescent molecules to the CNOs surfaces.
Latest applications of 3D ToF-SIMS bio-imaging
Authors: Fletcher John S.
Published: February 2015

Time-of-flight secondary ion mass spectrometry (ToF-SIMS) is a rapidly developing technique for the characterization of a wide range of materials. Recently, advances in instrumentation and sample preparation approaches have provided the ability to perform 3D molecular imaging experiments. Polyatomic ion beams, such as C60, and gas cluster ion beams, often Arn (n = 500–4000), substantially reduce the subsurface damage accumulation associated with continued bombardment of organic samples with atomic beams. In this review, the capabilities of the technique are discussed and examples of the 3D imaging approach for the analysis of model membrane systems, plant single cell, and tissue samples are presented. Ongoing challenges for 3D ToF-SIMS imaging are also discussed along with recent developments that might offer improved 3D imaging prospects in the near future.
First Report of Macrolide Resistance Gene
erm
(T) Harbored by a Novel Small Plasmid from Erysipelothrix rhusiopathiae
Authors: Xu Chang-Wen , Zhang An-Yun , Yang Chun-Mei , Pan Yun , Guan Zhong-Bin , Lei Chang-Wei , Peng Lin-Yao , Li Qing-Zhou , Wang Hong-Ning
Published: February 2015

The macrolide resistance gene
erm
(T) was identified for the first time in a porcine
Erysipelothrix rhusiopathiae
isolate from swine in China. The novel 3,749-bp small plasmid pER29, which carries
erm
(T), had a G+C content of 31% and four distinct open reading frames. The presence of pER29 increased by at least 128-fold the MICs of clindamycin and erythromycin for
E. rhusiopathiae
. The fitness cost of pER29 could be responsible for the low frequency of
erm
(T) in
E. rhusiopathiae
.
Factors Influencing Culture Positivity in Pyogenic Vertebral Osteomyelitis Patients with Prior Antibiotic Exposure
Authors: Kim Chung-Jong , Kang Seung-Ji , Yoon Doran , Lee Myung Jin , Kim Moonsuk , Song Kyoung-Ho , Jang Hee-Chang , Jung Sook-In , Kim Eu Suk , Kim Hong Bin , Oh Myoung-don , Park Kyung-Hwa , Kim Nam Joong
Published: February 2015

We conducted a retrospective cohort study to evaluate factors influencing tissue culture positivity in patients with pyogenic vertebral osteomyelitis exposed to antibiotics before diagnosis. Tissue culture was positive in 48.3% (28/58) of the patients, and the median antibiotic-free period was 1.5 days (range, 0.7 to 5.7 days). In a multivariate analysis, a higher C-reactive protein (CRP) level (adjusted odds ratio [aOR], 1.18; 95% confidence interval, 1.07 to 1.29) and open surgical biopsy (aOR, 6.33; 95% confidence interval, 1.12 to 35.86) were associated with tissue culture positivity.
Use of
In Vitro
Vancomycin Testing Results To Predict Susceptibility to Oritavancin, a New Long-Acting Lipoglycopeptide
Authors: Jones Ronald N. , Turnidge John D. , Moeck Greg , Arhin Francis F. , Mendes Rodrigo E.
Published: February 2015

Oritavancin is a recently approved lipoglycopeptide antimicrobial agent with activity against Gram-positive pathogens. Its extended serum elimination half-life and concentration-dependent killing enable single-dose treatment of acute bacterial skin and skin structure infections. At the time of regulatory approval, new agents, including oritavancin, are not offered in the most widely used susceptibility testing devices and therefore may require application of surrogate testing using a related antimicrobial to infer susceptibility. To evaluate vancomycin as a predictive susceptibility marker for oritavancin, 26,993 recent Gram-positive organisms from U.S. and European hospitals were tested using reference MIC methods. Organisms included
Staphylococcus aureus
, coagulase-negative staphylococci (CoNS), beta-hemolytic streptococci (BHS), viridans group streptococci (VGS), and enterococci (ENT). These five major pathogen groups were analyzed by comparing results with FDA-approved susceptible breakpoints for both drugs, as well as those suggested by epidemiological cutoff values and supported by pharmacokinetic/pharmacodynamic analyses. Vancomycin susceptibility was highly accurate (98.1 to 100.0%) as a surrogate for oritavancin susceptibility among the indicated pathogen species. Furthermore, direct MIC comparisons showed high oritavancin potencies, with vancomycin/oritavancin MIC
90
results of 1/0.06, 2/0.06, 0.5/0.12,1/0.06, and >16/0.06 μg/ml for
S. aureus
, CoNS, BHS, VGS, and ENT, respectively. In conclusion, vancomycin demonstrated acceptable accuracy as a surrogate marker for predicting oritavancin susceptibility when tested against the indicated pathogens. In contrast, 93.3% of vancomycin-nonsusceptible enterococci had oritavancin MIC values of ≤0.12 μg/ml, indicating a poor predictive value of vancomycin for oritavancin resistance against these organisms. Until commercial oritavancin susceptibility testing devices are readily available, isolates that when tested show vancomycin susceptibility can be inferred to be susceptible to oritavancin by using FDA-approved breakpoints.
Emergence of the First Levofloxacin-Resistant Strains of Streptococcus agalactiae Isolated in Italy
Authors: Piccinelli G. , Gargiulo F. , Corbellini S. , Ravizzola G. , Bonfanti C. , Caruso A. , De Francesco M. A.
Published: February 2015

Of 901 group B streptococcus strains analyzed, 13 (1.4%) were resistant to levofloxacin (MICs of >32 μg/ml for seven isolates, 2 μg/ml for four isolates, and 1.5 μg/ml for four isolates). Mutations in the quinolone resistance-determining regions (QRDRs) of gyrase and topoisomerase IV were identified. A double mutation involving the Ser-81 change to Leu for
gyrA
and the Ser-79 change to Phe or to Tyr for
parC
was linked to a high level of fluoroquinolone resistance. In addition, two other mutational positions in
parC
were observed, resulting in an Asp-83-to-Tyr substitution and an Asp-83-to-Asn substitution. Different mutations were also observed in
gyrB
, with unknown significance. Most levofloxacin-resistant GBS strains were of serotype Ib and belonged to sequence type 19 (ST19) and clonal complex 19 (CC-19). Most of them exhibited the
epsilon
gene.
Antimicrobial Peptides Expressed in Medicinal Maggots of the Blow Fly Lucilia sericata Show Combinatorial Activity against Bacteria
Authors: Pöppel Anne-Kathrin , Vogel Heiko , Wiesner Jochen , Vilcinskas Andreas
Published: February 2015

The larvae of the common green bottle fly (
Lucilia sericata
) produce antibacterial secretions that have a therapeutic effect on chronic and nonhealing wounds. Recent developments in insect biotechnology have made it possible to use these larvae as a source of novel anti-infectives. Here, we report the application of next-generation RNA sequencing (RNA-Seq) to characterize the transcriptomes of the larval glands, crop, and gut, which contribute to the synthesis of antimicrobial peptides (AMPs) and proteins secreted into wounds. Our data confirm that
L. sericata
larvae have adapted in order to colonize microbially contaminated habitats, such as carrion and necrotic wounds, and are protected against infection by a diverse spectrum of AMPs.
L. sericata
AMPs include not only lucifensin and lucimycin but also novel attacins, cecropins, diptericins, proline-rich peptides, and sarcotoxins. We identified 47 genes encoding putative AMPs and produced 23 as synthetic analogs, among which some displayed activities against a broad spectrum of microbial pathogens, including
Pseudomonas aeruginosa
,
Proteus vulgaris
, and
Enterococcus faecalis
. Against
Escherichia coli
(Gram negative) and
Micrococcus luteus
(Gram positive), we found mostly additive effects but also synergistic activity when selected AMPs were tested in combination. The AMPs that are easy to synthesize are currently being produced in bulk to allow their evaluation as novel anti-infectives that can be formulated in hydrogels to produce therapeutic wound dressings and adhesive bandages.
Interstrain Interactions between Bacteria Isolated from Vacuum-Packaged Refrigerated Beef
Authors: Zhang Peipei , Baranyi József , Tamplin Mark
Published: February 2015

The formation of bacterial spoilage communities in food is influenced by both extrinsic and intrinsic environmental factors. Although many reports describe how these factors affect bacterial growth, much less is known about interactions among bacteria, which may influence community structure. This study investigated interactions among representative species of bacteria isolated from vacuum-packaged (VP) beef. Thirty-nine effectors and 20 target isolates were selected, representing 10 bacterial genera:
Carnobacterium
,
Pseudomonas
,
Hafnia
,
Serratia
,
Yersinia
,
Rahnella
,
Brochothrix
,
Bacillus
,
Leuconostoc
, and
Staphylococcus
. The influence of live effectors on growth of target isolates was measured by spot-lawn agar assay and also in liquid culture medium broth using live targets and effector cell-free supernatants. Inhibition on agar was quantified by diameter of inhibition zone and in broth by measuring detection time, growth rate, and maximum population density. A number of interactions were observed, with 28.6% of isolates inhibiting and 4.2% promoting growth. The majority of
Pseudomonas
isolates antagonized growth of approximately one-half of target isolates. Two
Bacillus
spp. each inhibited 16 targets. Among lactic acid bacteria (LAB),
Carnobacterium maltaromaticum
inhibited a wider range of isolates compared to other LAB. The majority of effector isolates enhancing target isolate growth were Gram-negative, including
Pseudomonas
spp. and
Enterobacteriaceae
. These findings markedly improve the understanding of potential interactions among spoilage bacteria, possibly leading to more mechanistic descriptions of bacterial community formation in VP beef and other foods.
The Importance of the Pseudomonas aeruginosa Type III Secretion System in Epithelium Traversal Depends upon Conditions of Host Susceptibility
Authors: Sullivan Aaron B. , Tam K. P. Connie , Metruccio Matteo M. E. , Evans David J. , Fleiszig Suzanne M. J.
Published: February 2015

Pseudomonas aeruginosa
is invasive or cytotoxic to host cells, depending on the type III secretion system (T3SS) effectors encoded. While the T3SS is known to be involved in disease
in vivo
, how it participates remains to be clarified. Here, mouse models of superficial epithelial injury (tissue paper blotting with EGTA treatment) and immunocompromise (MyD88 deficiency) were used to study the contribution of the T3SS transcriptional activator ExsA to epithelial traversal. Corneas of excised eyeballs were inoculated with green fluorescent protein (GFP)-expressing PAO1 or isogenic
exsA
mutants for 6 h
ex vivo
before bacterial traversal and epithelial thickness were quantified by using imaging. In the blotting-EGTA model,
exsA
mutants were defective in capacity for traversal. Accordingly, an ∼16-fold variability in
exsA
expression among PAO1 isolates from three sources correlated with epithelial loss. In contrast, MyD88
−/−
epithelia remained susceptible to
P. aeruginosa
traversal despite
exsA
mutation. Epithelial lysates from MyD88
−/−
mice had reduced antimicrobial activity compared to those from wild-type mice with and without prior antigen challenge, particularly 30- to 100-kDa fractions, for which mass spectrometry revealed multiple differences, including (i) lower baseline levels of histones, tubulin, and lumican and (ii) reduced glutathione
S
-transferase, annexin, and dermatopontin, after antigen challenge. Thus, the importance of ExsA in epithelial traversal by invasive
P. aeruginosa
depends on the compromise enabling susceptibility, suggesting that strategies for preventing infection will need to extend beyond targeting the T3SS. The data also highlight the importance of mimicking conditions allowing susceptibility in animal models and the need to monitor variability among bacterial isolates from different sources, even for the same strain.
Molecular Analysis of Asymptomatic Bacteriuria Escherichia coli Strain VR50 Reveals Adaptation to the Urinary Tract by Gene Acquisition
Authors: Beatson Scott A. , Ben Zakour Nouri L. , Totsika Makrina , Forde Brian M. , Watts Rebecca E. , Mabbett Amanda N. , Szubert Jan M. , Sarkar Sohinee , Phan Minh-Duy , Peters Kate M. , Petty Nicola K. , Alikhan Nabil-Fareed , Sullivan Mitchell J. , Gawthorne Jayde A. , Stanton-Cook Mitchell , Nhu Nguyen Thi Khanh , Chong Teik Min , Yin Wai-Fong , Chan Kok-Gan , Hancock Viktoria , Ussery David W. , Ulett Glen C. , Schembri Mark A.
Published: February 2015

Urinary tract infections (UTIs) are among the most common infectious diseases of humans, with
Escherichia coli
responsible for >80% of all cases. One extreme of UTI is asymptomatic bacteriuria (ABU), which occurs as an asymptomatic carrier state that resembles commensalism. To understand the evolution and molecular mechanisms that underpin ABU, the genome of the ABU
E. coli
strain VR50 was sequenced. Analysis of the complete genome indicated that it most resembles
E. coli
K-12, with the addition of a 94-kb genomic island (GI-VR50-
pheV
), eight prophages, and multiple plasmids. GI-VR50-
pheV
has a mosaic structure and contains genes encoding a number of UTI-associated virulence factors, namely, Afa (afimbrial adhesin), two autotransporter proteins (Ag43 and Sat), and aerobactin. We demonstrated that the presence of this island in VR50 confers its ability to colonize the murine bladder, as a VR50 mutant with GI-VR50-
pheV
deleted was attenuated in a mouse model of UTI
in vivo
. We established that Afa is the island-encoded factor responsible for this phenotype using two independent deletion (Afa operon and AfaE adhesin) mutants.
E. coli
VR50
afa
and VR50
afaE
displayed significantly decreased ability to adhere to human bladder epithelial cells. In the mouse model of UTI, VR50
afa
and VR50
afaE
displayed reduced bladder colonization compared to wild-type VR50, similar to the colonization level of the GI-VR50-
pheV
mutant. Our study suggests that
E. coli
VR50 is a commensal-like strain that has acquired fitness factors that facilitate colonization of the human bladder.
Toll-Like Receptor 2 (TLR2) and TLR9 Play Opposing Roles in Host Innate Immunity against Salmonella enterica Serovar Typhimurium Infection
Authors: Zhan Renhui , Han Qiuju , Zhang Cai , Tian Zhigang , Zhang Jian
Published: February 2015

Toll-like receptors (TLRs) are evolutionarily conserved host proteins that are essential for effective host defense against pathogens. However, recent studies suggest that some TLRs can negatively regulate immune responses. We observed here that TLR2 and TLR9 played opposite roles in regulating innate immunity against oral infection ofSalmonella entericaserovar Typhimurium in mice. WhileTLR9−/−mice exhibited shortened survival, an increased cytokine storm, and more severeSalmonellahepatitis than wild-type (WT) mice,TLR2−/−mice exhibited the opposite phenomenon. Further studies demonstrated that TLR2 deficiency and TLR9 deficiency in macrophages both disrupted NK cell cytotoxicity againstS. Typhimurium-infected macrophages by downregulating NK cell degranulation and gamma interferon (IFN-γ) production through decreased macrophage expression of the RAE-1 NKG2D ligand. But more importantly, we found thatS. Typhimurium-infectedTLR2−/−macrophages upregulated inducible nitric oxide synthase (iNOS) expression, resulting in a lower bacterial load than that in WT macrophagesin vitroand liversin vivoas well as low proinflammatory cytokine levels. In contrast,TLR9−/−macrophages showed decreased reactive oxygen species (ROS) expression concomitant with a high bacterial load in the macrophages and in livers ofTLR9−/−mice.TLR9−/−macrophages were also more susceptible than WT macrophages toS. Typhimurium-induced necroptosisin vitro, likely contributing to bacterial spread and transmissionin vivo. Collectively, these findings indicate that TLR2 negatively regulates anti-S. Typhimurium immunity, whereas TLR9 is vital to host defense and survival againstS. Typhimurium invasion. TLR2 antagonists or TLR9 agonists may thus serve as potential anti-S. Typhimurium therapeutic agents.
The Class II Phosphatidylinositol 3-Phosphate Kinase PIK3C2A Promotes Shigella flexneri Dissemination through Formation of Vacuole-Like Protrusions
Authors: Dragoi Ana-Maria , Agaisse Hervé
Published: February 2015

Intracellular pathogens such asShigella flexneriandListeria monocytogenesachieve dissemination in the intestinal epithelium by displaying actin-based motility in the cytosol of infected cells. As they reach the cell periphery, motile bacteria form plasma membrane protrusions that resolve into vacuoles in adjacent cells, through a poorly understood mechanism. Here, we report on the role of the class II phosphatidylinositol 3-phosphate kinase PIK3C2A inS. flexneridissemination. Time-lapse microscopy revealed that PIK3C2A was required for the resolution of protrusions into vacuoles through the formation of an intermediate membrane-bound compartment that we refer to as a vacuole-like protrusion (VLP). Genetic rescue of PIK3C2A depletion with RNA interference (RNAi)-resistant cDNA constructs demonstrated that VLP formation required the activity of PIK3C2A in primary infected cells. PIK3C2A expression was required for production of phosphatidylinositol 3-phosphate [PtdIns(3)P] at the plasma membrane surrounding protrusions. PtdIns(3)P production was not observed in the protrusions formed byL. monocytogenes, whose dissemination did not rely on PIK3C2A. PIK3C2A-mediated PtdIns(3)P production inS. flexneriprotrusions was regulated by host cell tyrosine kinase signaling and relied on the integrity of theS. flexneritype 3 secretion system (T3SS). We suggest a model ofS. flexneridissemination in which the formation of VLPs is mediated by the PIK3C2A-dependent production of the signaling lipid PtdIns(3)P in the protrusion membrane, which relies on the T3SS-dependent activation of tyrosine kinase signaling in protrusions.
Analysis of the σ
E
Regulon in Crohn's Disease-Associated Escherichia coli Revealed Involvement of the
waaWVL
Operon in Biofilm Formation
Authors: Chassaing Benoit , Garénaux Estelle , Carriere Jessica , Rolhion Nathalie , Guérardel Yann , Barnich Nicolas , Bonnet Richard , Darfeuille-Michaud Arlette
Published: February 2015

Ileal lesions of patients with Crohn's disease are colonized by adherent-invasive
Escherichia coli
(AIEC), which is able to adhere to and to invade intestinal epithelial cells (IEC), to replicate within macrophages, and to form biofilms on the surface of the intestinal mucosa. Previous analyses indicated the involvement of the σ
E
pathway in AIEC-IEC interaction, as well as in biofilm formation, with σ
E
pathway inhibition leading to an impaired ability of AIEC to colonize the intestinal mucosa and to form biofilms. The aim of this study was to characterize the σ
E
regulon of AIEC strain LF82 in order to identify members involved in AIEC phenotypes. Using comparative
in silico
analysis of the σ
E
regulon, we identified the
waaWVL
operon as a new member of the σ
E
regulon in reference AIEC strain LF82. We determined that the
waaWVL
operon is involved in AIEC lipopolysaccharide structure and composition, and the
waaWVL
operon was found to be essential for AIEC strains to produce biofilm and to colonize the intestinal mucosa.

IMPORTANCE
An increased prevalence of adherent-invasive
Escherichia coli
(AIEC) bacteria was previously observed in the intestinal mucosa of Crohn's disease (CD) patients, and clinical observations revealed bacterial biofilms associated with the mucosa of CD patients. Here, analysis of the σ
E
regulon in AIEC and commensal
E. coli
identified 12 genes controlled by σ
E
only in AIEC. Among them, WaaWVL factors were found to play an essential role in biofilm formation and mucosal colonization by AIEC. In addition to identifying molecular tools that revealed a pathogenic population of
E. coli
colonizing the mucosa of CD patients, these results indicate that targeting the
waaWVL
operon could be a potent therapeutic strategy to interfere with the ability of AIEC to form biofilms and to colonize the gut mucosa.
Genome-Wide Analysis of Phosphorylated PhoP Binding to Chromosomal DNA Reveals Several Novel Features of the PhoPR-Mediated Phosphate Limitation Response in Bacillus subtilis
Authors: Salzberg Letal I. , Botella Eric , Hokamp Karsten , Antelmann Haike , Maaß Sandra , Becher Dörte , Noone David , Devine Kevin M.
Published: February 2015

The PhoPR two-component signal transduction system controls one of three responses activated by
Bacillus subtilis
to adapt to phosphate-limiting conditions (PHO response). The response involves the production of enzymes and transporters that scavenge for phosphate in the environment and assimilate it into the cell. However, in
B. subtilis
and some other
Firmicutes
bacteria, cell wall metabolism is also part of the PHO response due to the high phosphate content of the teichoic acids attached either to peptidoglycan (wall teichoic acid) or to the cytoplasmic membrane (lipoteichoic acid). Prompted by our observation that the phosphorylated WalR (WalR∼P) response regulator binds to more chromosomal loci than are revealed by transcriptome analysis, we established the PhoP∼P bindome in phosphate-limited cells. Here, we show that PhoP∼P binds to the chromosome at 25 loci: 12 are within the promoters of previously identified PhoPR regulon genes, while 13 are newly identified. We extend the role of PhoPR in cell wall metabolism showing that PhoP∼P binds to the promoters of four cell wall-associated operons (
ggaAB
,
yqgS
,
wapA
, and
dacA
), although none show PhoPR-dependent expression under the conditions of this study. We also show that positive autoregulation of
phoPR
expression and full induction of the PHO response upon phosphate limitation require PhoP∼P binding to the 3′ end of the
phoPR
operon.

IMPORTANCE
The PhoPR two-component system controls one of three responses mounted by
B. subtilis
to adapt to phosphate limitation (PHO response). Here, establishment of the phosphorylated PhoP (PhoP∼P) bindome enhances our understanding of the PHO response in two important ways. First, PhoPR plays a more extensive role in adaptation to phosphate-limiting conditions than was deduced from transcriptome analyses. Among 13 newly identified binding sites, 4 are cell wall associated (
ggaAB
,
yqgS
,
wapA
, and
dacA
), revealing that PhoPR has an extended involvement in cell wall metabolism. Second, amplification of the PHO response must occur by a novel mechanism since positive autoregulation of
phoPR
expression requires PhoP∼P binding to the 3′ end of the operon.
Role of RNA Secondary Structure and Processing in Stability of the
nifH1
Transcript in the Cyanobacterium Anabaena variabilis
Authors: Pratte Brenda S. , Ungerer Justin , Thiel Teresa
Published: February 2015

In the cyanobacterium
Anabaena variabilis
ATCC 29413, aerobic nitrogen fixation occurs in micro-oxic cells called heterocysts. Synthesis of nitrogenase in heterocysts requires expression of the large
nif1
gene cluster, which is primarily under the control of the promoter for the first gene,
nifB1
. Strong expression of
nifH1
requires the
nifB1
promoter but is also controlled by RNA processing, which leads to increased
nifH1
transcript stability. The processing of the primary
nifH1
transcript occurs at the base of a predicted stem-loop structure that is conserved in many heterocystous cyanobacteria. Mutations that changed the predicted secondary structure or changed the sequence of the stem-loop had detrimental effects on the amount of
nifH1
transcript, with mutations that altered or destabilized the structure having the strongest effect. Just upstream from the transcriptional processing site for
nifH1
was the promoter for a small antisense RNA,
sava4870.1
. This RNA was more strongly expressed in cells grown in the presence of fixed nitrogen and was downregulated in cells 24 h after nitrogen step down. A mutant strain lacking the promoter for
sava4870.1
showed delayed nitrogen fixation; however, that phenotype might have resulted from an effect of the mutation on the processing of the
nifH1
transcript. The
nifH1
transcript was the most abundant and most stable
nif1
transcript, while
nifD1
and
nifK1
, just downstream of
nifH1
, were present in much smaller amounts and were less stable. The
nifD1
and
nifK1
transcripts were also processed at sites just upstream of
nifD1
and
nifK1
.

IMPORTANCE
In the filamentous cyanobacterium
Anabaena variabilis
, the
nif1
cluster, encoding the primary Mo nitrogenase, functions under aerobic growth conditions in specialized cells called heterocysts that develop in response to starvation for fixed nitrogen. The large cluster comprising more than a dozen
nif1
genes is transcribed primarily from the promoter for the first gene,
nifB1
; however, this does not explain the large amount of transcript for the structural genes
nifH1
,
nifD1
, and
nifK1
, which are also under the control of the distant
nifB1
promoter. Here, we demonstrate the importance of a predicted stem-loop structure upstream of
nifH1
that controls the abundance of
nifH1
transcript through transcript processing and stabilization and show that
nifD1
and
nifK1
transcripts are also controlled by transcript processing.
‘Pseudotumour’ invading the proximal femur with normal metal ions following metal on metal hip resurfacing
Authors: Krishnan Harry , Sugand Kapil , Ali Ibrahim , Smith Jay
Published: February 2015

A 75-year-old woman who had undergone hybrid metal-on-metal hip resurfacing 8 years earlier underwent revision arthroplasty because of hip, groin and lateral thigh pain. The main differential was aseptic loosening; however, serum cobalt and chromium levels were normal. Multiple imaging modalities revealed a periprosthetic, cystic soft tissue mass adjacent to the proximal femur. A large ‘pseudotumour’ with proximal femoral invasion was found at revision arthroplasty. We report the first finding of a ‘pseudotumour’ invading the proximal femur with normal metal ions following metal on metal hip resurfacing.
Cystic meningioma
Authors: Amin Osama S M , Shwani Saad Suud , Khalifa Firas
Published: February 2015

Cystic meningioma is an uncommon form of meningioma and the radiological appearance and location of the cystic/solid components of the mass may create a diagnostic dilemma. We report on the case of an 80-year-old left-handed man who presented with recurrent focal motor seizures and secondary generalisation. A large left-sided parieto-occipital cystic mass was detected on contrast CT of the brain. The appearance suggested a malignant tumour. However, brain MRI with gadolinium delineated the solid and cystic component precisely. A provisional diagnosis of cystic meningioma was made and this was confirmed histologically after resecting the tumour surgically. It was a benign WHO Grade I fibroblastic meningioma. The preoperative diagnosis of cystic meningioma is not that straightforward. Brain MRI with gadolinium has a better diagnostic yield than CT scanning. Histopathological examination of the tumour cells should always be performed to confirm the category and subtype of the tumour.
Patients’ experiences of the choice of GP practice pilot, 2012/2013: a mixed methods evaluation
Authors: Tan Stefanie , Erens Bob , Wright Michael , Mays Nicholas
Published: February 2015

Objectives

To investigate patients’ experiences of the choice of general practitioner (GP) practice pilot.

Design

Mixed-method, cross-sectional study.

Setting

Patients in the UK National Health Service (NHS) register with a general practice responsible for their primary medical care and practices set geographic boundaries. In 2012/2013, 43 volunteer general practices in four English NHS primary care trusts (PCTs) piloted a scheme allowing patients living outside practice boundaries to register as an out of area patient or be seen as a day patient.

Participants

Analysis of routine data for 1108 out of area registered patients and 250 day patients; postal survey of out of area registered (315/886, 36%) and day (64/188, 34%) patients over 18 years of age, with a UK mailing address; comparison with General Practice Patient Survey (GPPS); semistructured interviews with 24 pilot patients.

Results

Pilot patients were younger and more likely to be working than non-pilot patients at the same practices and reported generally more or at least as positive experiences than patients registered at the same practices, practices in the same PCT and nationally, despite belonging to subgroups of the population who typically report poorer than average experiences. Out of area patients who joined a pilot practice did so: after moving house and not wanting to change practice (26.2%); for convenience (32.6%); as newcomers to an area who selected a practice although they lived outside its boundary (23.6%); because of dissatisfaction with their previous practice (13.9%). Day patients attended primarily on grounds of convenience (68.8%); 51.6% of the day patient visits were for acute infections, most commonly upper respiratory infections (20.4%). Sixty-six per cent of day patients received a prescription during their visit.

Conclusions

Though the 12-month pilot was too brief to identify all costs and benefits, the scheme provided a positive experience for participating patients and practices.
Zonal T2* and T1Gdassessment of knee joint cartilage in various histological grades of cartilage degeneration: an observational in vitro study
Authors: Bittersohl Bernd , Hosalkar Harish S , Miese Falk R , Schibensky Jonas , König Dietmar P , Herten Monika , Antoch Gerald , Krauspe Rüdiger , Zilkens Christoph
Published: February 2015

Objectives

Accurate assessment of cartilage status is increasingly becoming important to clinicians for offering joint preservation surgeries versus joint replacements. The goal of this study was to evaluate the validity of three-dimensional (3D), gradient-echo (GRE)-based T2* and T1Gdmapping for the assessment of various histological severities of degeneration in knee joint cartilage with potential implications for clinical management.

Methods

MRI and histological assessment were conducted in 36 ex vivo lateral femoral condyle specimens. The MRI protocol included a 3D GRE multiecho data image combination sequence in order to assess the T2* decay, a 3D double-echo steady-state sequence for assessment of cartilage morphology, and a dual flip angle 3D GRE sequence with volumetric interpolated breathhold examination for the T1Gdassessment. The histological sample analysis was performed according to the Mankin system. The data were then analysed statistically and correlated.

Results

We observed a significant decrease in the T2* and T1Gdvalues with increasing grades of cartilage degeneration (p<0.001) and a moderate correlation between T2* (r=0.514)/T1Gd(r=0.556) and the histological grading of cartilage degeneration (p<0.001). In addition, we noted a zonal variation in the T2* and T1Gdvalues reflecting characteristic zonal differences in the biochemical composition of hyaline cartilage.

Conclusions

This study outlines the potential of GRE-based T2* and T1Gdmapping to identify various grades of cartilage damage. Early changes in specific zones may assist clinicians in identifying methods of early intervention involving the targeted joint preservation approach versus moving forward with unicompartmental, bicompartmental or tricompartmental joint replacement procedures.

Trial registration number

DRKS00000729.
Effectiveness of a multifactorial intervention on preventing development of frailty in pre-frail older people: study protocol for a randomised controlled trial
Authors: Fairhall Nicola , Kurrle Susan E , Sherrington Catherine , Lord Stephen R , Lockwood Keri , John Beatrice , Monaghan Noeline , Howard Kirsten , Cameron Ian D
Published: February 2015

Introduction

Frailty is a major concern due to its costly and widespread consequences, yet evidence of effective interventions to delay or reduce frailty is lacking. Our previous study found that a multifactorial intervention was feasible and effective in reducing frailty in older people who were already frail. Identifying and treating people in the pre-frail state may be an effective means to prevent or delay frailty. This study describes a randomised controlled trial that aims to evaluate the effectiveness of a multifactorial intervention on development of frailty in older people who are pre-frail.

Methods and analysis

A single centre randomised controlled trial with concealed allocation, assessor blinding and intention-to-treat analysis. Two hundred and thirty people aged above 70 who meet the Cardiovascular Health Study frailty criteria for pre-frailty, reside in the community and are without severe cognitive impairment will be recruited. Participants will be randomised to receive a multifactorial intervention or usual care. The intervention group will receive a 12-month interdisciplinary intervention targeting identified characteristics of frailty and problems identified during geriatric assessment. Participants will be followed for a 12-month period. Primary outcome measures will be degree of frailty measured by the number of Cardiovascular Health Study frailty criteria present, and mobility measured with the Short Physical Performance Battery. Secondary outcomes will include measures of mobility, mood and use of health and community services.

Ethics and dissemination

The study was approved by the Northern Sydney Local Health District Health Research Ethics Committee (1207-213M). The findings will be disseminated through scientific and professional conferences, and in peer-reviewed journals.

Trial registration number

Australian New Zealand Clinical Trials Registry: ACTRN12613000043730.
Promoting vision and hearing aids use in an intensive care unit
Authors: Zhou Qiaoling , Faure Walker Nicholas
Published: February 2015

Vision and hearing impairments have long been recognised as modifiable risk factors for delirium.[1,2,3] Delirium in critically ill patients is a frequent complication (reported as high as 60% to 80% of intensive care patients), and is associated with a three-fold increase in mortality and prolonged hospital stay.[1] Guidelines by the UK Clinical Pharmacy Association recommend minimising risk factors to prevent delirium, rather than to treat it with pharmacological agents which may themselves cause delirium.[4] To address risk factors is a measure of multi-system management, such as sleep-wake cycle correction, orientation and use of vision and hearing aids, etc.[5]

We designed an audit to survey the prevalence and availability of vision and hearing aids use in the intensive care unit (ICU) of one university hospital. The baseline data demonstrated a high level of prevalence and low level of availability of vision /hearing aid use.

We implemented changes to the ICU Innovian assessment system, which serves to remind nursing staff performing daily checks on delirium reduction measures. This has improved practice in promoting vision and hearing aids use in ICU as shown by re-audit at six month. Further amendments to the Innovian risk assessments have increased the rate of assessment to 100% and vision aid use to near 100%.
Training veterinarians and agricultural advisers on a novel tool for tail biting prevention
Authors: vom Brocke A. L. , Madey D. P. , Gauly M. , Schrader L. , Dippel S.
Published: February 2015

Introduction

Many health and welfare problems in modern livestock production are multifactorial problems which require innovative solutions, such as novel risk assessment and management tools. However, the best way to distribute such novel ‐ and usually complex ‐ tools to the key applicants still has to be discussed.

Materials and methods

This paper shares experiences from distributing a novel tail biting prevention tool (‘SchwIP’) to 115 farm advisers and 19 veterinarians in 23 one‐day workshops. Participants gave written and oral feedback at the end of the workshops, which was later analysed together with the number of farms they had visited after the workshops. Workshop groups were categorised into groups showing (a) HIGH, (b) INTermediate or (c) LOW levels of antagonism against SchwIP or parts of it during workshop discussions.

Results

Group types did not significantly differ in their evaluation of knowledge transfer. However, HIGH group members evaluated the on‐farm usability of the tool significantly lower in the workshop feedback and tended to visit fewer farms.

Conclusions

As antagonistic discussion can influence workshop output, future workshop leaders should strive for basic communication training as well as some group leadership experience before setting up and leading workshops.
Influence of dog appeasing pheromone (DAP) on dogs housed in a long‐term kennelling facility
Authors: Grigg E. K. , Piehler M.
Published: February 2015

Introduction

Kennel facilities are commonly acknowledged as a stressful environment for many domestic dogs (Canis familiaris). One therapeutic measure used to reduce anxiety in dogs is dog appeasing pheromone (DAP), which has been found effective in reducing stress‐related behaviours in a number of contexts.

Aims and Objectives

A pilot study was conducted to assess whether DAP would reduce frequency of stress‐related behaviours in a group of eight dogs housed for teaching purposes in a long‐term kennelling facility.

Materials and Methods

Using video analysis, proportion of time spent in stress‐related behaviours for six dogs fitted with DAP collars, versus two control dogs (without collars), was compared for the time before and during DAP exposure.

Results

No significant differences were found either in the proportion of time spent in stress‐related behaviours in the baseline versus treatment periods or between the collared and control dogs in the change in proportion of time they spent in any of the focal behaviours in the baseline versus treatment periods.

Conclusions

Possible reasons for these findings include an actual lack of effect of DAP on dogs housed in this long‐term kennelling facility, an apparent lack of effect due to small sample size in this pilot study and high behavioural variation among individual dogs. Despite lack of a demonstrated effect of the DAP collars on these dogs, attention brought by this study to the behavioural issues seen in some of the dogs did have a positive impact, as it contributed to the development of an active, coordinated behavioural wellness and enrichment programme for the colony.