bluuebunny/crossref_metadata_2025 huggingface dataset:
Development and Evaluation of an ELISA for the Quantitation of Anti‐Lagenidium giganteum forma caninum Antibodies in Dogs
Authors: Hartfield J.N. , Grooters A.M. , Waite K.J.
Published: October 2014

Background

Lagenidium giganteum forma caninum infection causes severe cutaneous and disseminated disease in dogs. Currently, diagnosis requires culture and rRNA gene sequencing.

Objective

To develop and evaluate an ELISA for quantitation of anti‐L. giganteum f. caninum IgG in canine serum.

Animals

Sera were evaluated from 22 dogs infected with L. giganteum f. caninum, 12 dogs infected with Paralagenidium karlingii, 18 dogs infected with Pythium insidiosum, 26 dogs with nonoomycotic fungal infections or other cutaneous or systemic diseases, and 10 healthy dogs.

Methods

Antigen was prepared from a soluble mycelial extract of L. giganteum f. caninum. Optimal antigen and antibody concentrations were determined by checkerboard titration. Results were expressed as percent positivity (PP) relative to a strongly positive control serum.

Results

Medians and ranges for PP for each group were: L. giganteum f. caninum (73.9%, 27.9–108.9%), P. karlingii (55.0%, 21.0–90.6%), P. insidiosum (31.3%, 15.8–87.5%), nonoomycotic fungal infection or other cutaneous or systemic diseases (19.2%, 3.2–61.0%), and healthy dogs (9.9%, 7.6–24.6%). Using a PP cutoff value of 40%, sensitivity and specificity (with 95% CI) of the ELISA for detecting L. giganteum f. caninum infection in clinically affected dogs were 90.9% (72.2–97.5%) and 73.2% (60.4–83.0%), respectively. Specificity in dogs infected with P. karlingii was 41.7% (19.3–68.1%) and with P. insidiosum was 66.7% (43.8–83.7%).

Conclusions and Clinical Importance

Quantitation of anti‐L. giganteum f. caninum antibodies for detection of this infection in dogs has moderately high sensitivity but poor specificity, the latter because of substantial cross‐reactivity with anti‐P. karlingii and anti‐P. insidiosum antibodies.
Pharmacokinetics of intravenous, plain oral and enteric‐coated oral omeprazole in the horse
Authors: Sykes B. W. , Underwood C. , McGowan C. M. , Mills P. C.
Published: October 2014

The objectives were to document the pharmacokinetics of intravenous, enteric‐coated oral and plain oral omeprazole in fasted horses and to investigate the impact of feeding on the bioavailability of an enteric‐coated omeprazole. Twelve horses received four treatments: intravenous omeprazole (0.5 mg/kg) in the fasted state (IV‐Fasted), enteric‐coated omeprazole (4 mg/kg) orally in the fasted state (ECO‐Fasted), enteric‐coated omeprazole (4 mg/kg) orally in the fed state (ECO‐Fed) and plain omeprazole (4 mg/kg) orally in the fasted state (PL‐Fasted). Plasma omeprazole concentrations were determined by UHPLC‐MS. Bioavailability was higher (P = 0.038) in the ECO‐Fasted group (21.5 [9.0–27.7]%) than the PL‐Fasted group (10.1 [7.7–13.3]%). Similarly, AUC0‐∞ was higher in the ECO‐Fasted group than the PL‐Fasted group (P = 0.027). No significant differences were present between the ECO‐Fasted and ECO‐Fed groups with regards to bioavailability, Cmax, Tmax or AUC0‐∞. When the half‐life data from the oral formulations was pooled, it was longer than that observed in the IV‐Fasted group (100 [73–118] min) and 35 [34‐39] min, respectively; P < 0.0001). Bioavailability of enteric‐coated omeprazole was higher than previously reported and feeding had minimal impact. Bioavailability of plain omeprazole was approximately half that of enteric‐coated omeprazole. The longer half‐life observed following oral administration was consistent with the flip‐flop effect and has not previously been described for omeprazole in the horse.
Job satisfaction of Malaysian registered nurses: a qualitative study
Authors: Atefi Narges , Abdullah Khatijah L , Wong Li P
Published: October 2014

Background

Job satisfaction is an important factor in health care settings. Strong empirical evidence supports a causal relationship between job satisfaction, patient safety and quality of care. However, there have not been any studies exploring the job satisfaction of Malaysian nurses.

Aim

The main purpose of this qualitative descriptive study was to explore the factors related to feelings of job satisfaction as well as job dissatisfaction experienced by registered nurses in Malaysia.

Method

A convenient sample of 46 Malaysian nurses recruited from a large hospital (number of beds = 895) participated in the study. A total of seven focus group discussions were conducted with nurses from surgical, medical and critical care wards. A semi‐structured interview guide was used to facilitate the interviews, which were audio‐recorded, transcribed verbatim and checked. The transcripts were used as data and were analysed using a thematic approach.

Finding

The study identified three main themes that influenced job satisfaction: (1) nurses' personal values and beliefs; (2) work environment factors and (3) motivation factors. Concerning the nurses' personal values and beliefs, the ability to help people made the nurses felt honoured and happy, which indirectly contributed to job satisfaction. For work environment factors, team cohesion, benefit and reward, working conditions play an important role in the nurses' job satisfaction. Motivation factors, namely, professional development and clinical autonomy contributed to job satisfaction.

Conclusion

It is important for nurse leaders to provide more rewards, comfortable work environments and to understand issues that affect nurses' job satisfaction.

Relevance to clinical practice

Our findings highlight the importance of factors that can improve nurses' job satisfaction. The study provides basic information for hospital administrators in planning effective and efficient policies to improve nursing job satisfaction in order to increase the quality of patient care and decrease nursing turnover.
Intervention to improve intensive care nurses' knowledge of sedation assessment and management
Authors: Ramoo Vimala , Abdullah Khatijah L , Tan Patrick SK , Wong Li P , Chua Piaw Y
Published: October 2014

Background

Sedation management is an integral component of critical care practice. It requires the greatest attention of critical care practitioners because it carries significant risks to patients. Therefore, it is imperative that nurses are aware of potential adverse consequences of sedation therapy and current sedation practice recommendations.

Aims and objectives

To evaluate the impact of an educational intervention on nurses' knowledge of sedation assessment and management.

Designs and methods

A quasi‐experimental design with a pre‐ and post‐test method was used. The educational intervention included theoretical sessions on assessing and managing sedation and hands‐on sedation assessment practice using the Richmond Agitation Sedation Scale. Its effect was measured using self‐administered questionnaire, completed at the baseline level and 3 months following the intervention.

Results

Participants were 68 registered nurses from an intensive care unit of a teaching hospital in Malaysia. Significant increases in overall mean knowledge scores were observed from pre‐ to post‐intervention phases (mean of 79·00 versus 102·00, p < 0·001). Nurses with fewer than 5 years of work experience, less than 26 years old, and with a only basic nursing education had significantly greater level of knowledge improvement at the post‐intervention phase compared to other colleagues, with mean differences of 24·64 (p = 0·001), 23·81 (p = 0·027) and 27·25 (p = 0·0001), respectively. A repeated‐measures analysis of variance revealed a statistically significant effect of educational intervention on knowledge score after controlling for age, years of work and level of nursing education (p = 0·0001, ηp2 = 0·431).

Conclusion

An educational intervention consisting of theoretical sessions and hands‐on sedation assessment practice was found effective in improving nurses' knowledge and understanding of sedation management.

Relevance to clinical practice

This study highlighted the importance of continuing education to increase nurses' understanding of intensive care practices, which is vital for improving the quality of patient care.
Masticatory Muscles Dysfunction after CT‐guided Percutaneous Trigeminal Radiofrequency Thermocoagulation for Trigeminal Neuralgia: A Detailed Analysis
Authors: Zheng Shuyue , Wu Baishan , Zhao Ying , Wang Xiaoyu , Li Xuanying , Yang Liqiang , He Mingwei , Yue Jianning , Ni Jiaxiang
Published: October 2014

Objective

The aim of this study was to investigate the severity and the natural course of masticatory muscles weakness that developed after CT‐guided percutaneous trigeminal radiofrequency thermocoagulation (PT‐RFT) for the treatment of idiopathic trigeminal neuralgia (ITN).

Methods

Twenty‐seven patients with ITN were treated by CT‐guided percutaneous trigeminal radiofrequency thermocoagulation. Each patients' occlusal function and surface electromyographic (sEMG) activity of the ipsilateral anterior temporalis (TA) and masseter muscles (MM) at mandibular postural position (MPP), and during a fast maximum voluntary clenching (MVC) from MPP to intercuspal position (ICP), were simultaneously recorded by the T‐Scan III system and Bio‐pak sEMG III system before (baseline), 3 days, 3 months, and 12 months after procedure. The incidence, degree, and prognosis of masticatory muscles dysfunction related to trigeminal nerve motor‐branch injury were analyzed.

Results

Three days and 3 months after procedure, both the occlusal symmetry and the sEMG activity of ipsilateral TA and MM became significantly decreased compared to the baseline (P < 0.05). However, they demonstrated a gradual improvement toward preoperative values in follow‐up, returning to complete in 23 patients at 12 months after procedure. None reported permanent masticatory paralysis. Pain relief was most significant on the third day after procedure. At the final clinical visit, a pain‐free status was observed in 25 patients (92.6%). Meanwhile, the intensity of facial dysesthesia was mildest, whereas there were statistic differences compared with baseline.

Conclusion

CT‐guided PT‐RFT for ITN remains an effective and safe surgical procedure, but there is a high rate of temporary masticatory dysfunction during a short time after procedure, appearing to be reversible in a period of 12 months.
A novel method for determination of the 15N isotopic composition of Rubisco in wheat plants exposed to elevated atmospheric carbon dioxide
Authors: Aranjuelo Iker , Molero Gemma , Avice Jean Christophe , Bourguignon Jacques
Published: October 2014

Although ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) is mostly known as a key enzyme involved in CO2 assimilation during the Calvin cycle, comparatively little is known about its role as a pool of nitrogen storage in leaves. For this purpose, we developed a protocol to purify Rubisco that enables later analysis of its 15N isotope composition (δ15N) at the natural abundance and 15N‐labeled plants. In order to test the utility of this protocol, durum wheat (Triticum durum var. Sula) exposed to an elevated CO2 concentration (700 vs 400 µmol mol−1) was labeled with K15NO3 (enriched at 2 atom %) during the ear development period. The developed protocol proves to be selective, simple, cost effective and reproducible. The study reveals that 15N labeling was different in total organic matter, total soluble protein and the Rubisco fraction. The obtained data suggest that photosynthetic acclimation in wheat is caused by Rubisco depletion. This depletion may be linked to preferential nitrogen remobilization from Rubisco toward grain filling.
Temperature modulation of thermal tolerance of a CAM‐tank bromeliad and the relationship with acid accumulation in different leaf regions
Authors: Chaves Cleber Juliano Neves , Leal Bárbara Simões Santos , de Lemos‐Filho José Pires
Published: October 2014

Physiological changes that increase plant performance during exposure to high temperatures may play an inverse role during exposure to low temperatures. The objective of this study was to test variations in photosystem II response to heat and cold stress in the leaves of a bromeliad with crassulacean acid metabolism submitted to high or low temperatures. Leaves were maintained under constant temperatures of 10 and 35°C and used to examine possible relationships among physiological responses to high and low temperatures and organic acid accumulation. We also tested if distinct parts of bromeliad leaves show differences in photosynthetic thermotolerance. The samples from leaves maintained at 35°C showed greater heat tolerance values, while those from leaves maintained at 10°C showed lower cold tolerance values. Our results identified a strong negative relationship between the organic acid accumulation and thermal tolerance of bromeliad leaves that largely explained the differences in thermal tolerance among groups. One of these differences occurred among regions of a single leaf, with the base showing critical heat values of up to 8°C higher than the top region, suggesting a possible partitioning of leaf response among its regions. Differences in thermal tolerance were also observed between sampling times, with higher values observed in the morning.
Temperament, Parenting, and Moral Development: Specificity of Behavior and Context
Authors: Augustine Mairin E. , Stifter Cynthia A.
Published: October 2014

This longitudinal study highlights the role of specific parenting behaviors in specific contexts when predicting moral development in children of varying temperament types. A sample of mother–child dyads took part in a competing demands task involving differing ‘do’ and ‘don't’ contextual demands when the child was 2 years of age. Child temperament was also assessed at this time, yielding inhibited, exuberant, and low‐reactive temperament groups. Children's moral behavior was assessed at 5.5 years of age. Models examining the interaction of temperament and mother behaviors in each context indicated that mother's reasoning/explanation and ignoring in the ‘do’ context predicted later moral behavior in inhibited children whereas redirection and commands in the ‘don't’ context predicted moral behavior in exuberant children.
Mothers' and Fathers' Emotion Socialization and Children's Emotion Regulation: A Within‐Family Model
Authors: Shewark Elizabeth A. , Blandon Alysia Y.
Published: October 2014

In the current study, we examined whether mothers' and fathers' reactions to young children's positive and negative emotions were associated with children's negativity and emotion regulation. We utilized a within‐family design with 70 families (mother, father, and two siblings between the ages of 2 and 5 years). Mothers and fathers completed questionnaires about their emotion socialization as well as children's negativity and emotion regulation. Results indicated that mothers' and fathers' unsupportive reactions to children's positive emotions were associated with children's negativity. Fathers' unsupportive reactions to children's emotional displays were differentially associated with older and younger siblings' emotion regulation. Fathers' unsupportive responses to children's positive and negative emotions also contributed jointly to children's emotion regulation. The results suggest that exploring the within‐family correlates of children's emotion regulation and negativity is useful for understanding children's emotional development.
STEM Teachers' Planned and Enacted Attempts at Implementing Engineering Design‐Based Instruction
Authors: Capobianco Brenda M. , Rupp Madeline
Published: October 2014

This study investigates grades 5 and 6 science, technology, engineering, and mathematics (STEM) teachers' planned and actualized engineering design‐based instruction, the instruments used to characterize their efforts, and the implications this work has for teachers' implementations of an integrated approach to STEM education. Participants included 23 STEM teachers from six schools (three rural, two suburban, and one urban). Data were gathered via lesson implementation plans and classroom observations. Teachers demonstrated strength in planning for standards‐ and engineering design‐based lessons, incorporating engineering practices within their respective implementation plans, and aligning their plans with content and design process standards. Missing from their plans was attention to science concepts and their placement, use, and application within a design task. Classroom observations indicated that the teacher participants gave priority to “front loading,” the design process by concentrating more of their instructional time on problem identification and planning and less time on testing designs, communicating performance results, and redesigning. Measures utilized in this study provided insight into the content of teachers' planning and subsequent instruction and suggest potential for capturing content planning in the context of classrooms in which teachers are attempting to integrate novel curriculum, such as the new standards for engineering practices.
The Effect of a Horseshoe Crab Citizen Science Program on Middle School Student Science Performance and STEM Career Motivation
Authors: Hiller Suzanne E. , Kitsantas Anastasia
Published: October 2014

The purpose of the present quasi‐experimental study was to examine the impact of a horseshoe crab citizen science program on student achievement and science, technology, engineering, and mathematics (STEM) career motivation with 86 (n = 86) eighth‐grade students. The treatment group conducted fieldwork with naturalists and collected data for a professional biologist studying horseshoe crab speciation and a mock survey. The comparison group studied curriculum related to horseshoe crabs in the science classroom. A series of measures related to self‐efficacy, interest, outcome expectations, choice goals, and content knowledge were given to participants before and after the intervention. It was hypothesized that students would report higher motivational beliefs regarding science and show higher levels of achievement following the intervention than the comparison group. Support was shown for most of the hypotheses. In addition, path analyses indicated that students' motivational beliefs influence content knowledge and outcome expectations, which in turn affect their career goals. These results have implications for incorporating authentic fieldwork within a formal school structure as an effective method for promoting student achievement and STEM career motivation.
The Effect of GATT/WTO on Export and Import Price Volatility
Authors: Cao Vinh T. H. , Flach Lisandra
Published: October 2014

Despite much interest and discussion concerning the trade‐promoting effects of membership in multilateral trade agreements, little is known about the effect of membership on world trade prices. This paper fills this gap by studying the effects of membership on export and import price volatility. We document a surprisingly strong and robust empirical regularity: GATT/WTO membership reduces the volatility of prices over time for both import and export countries, with similar results found for free trade agreements. We show that results are not driven by sample selection or endogeneity concerns and that the effect is captured by members subject to rigorous accession procedures.
A pulse‐dose topical 1% 5‐fluorouracil treatment regimen in a young dog with corneal squamous cell carcinoma
Authors: Overton Taryn L. , Allbaugh Rachel A. , Whitley David , Ben‐Shlomo Gil , Griggs Angel , Tofflemire Kyle L. , Whitley Elizabeth M.
Published: October 2014

Purpose

To describe the use of a pulse‐dose topical 5‐fluorouracil (5‐FU) treatment regimen in a Pug dog with corneal squamous cell carcinoma (SCC).

Methods

A 1‐year‐old, spayed female Pug was evaluated for a corneal perforation of the right eye, which was surgically stabilized with a conjunctival pedicle graft. At the time of medial canthoplasty 7 weeks later, two areas of gray–white discoloration had developed medial and lateral to the graft. Biopsy samples were obtained via superficial keratectomy while under general anesthesia.

Results

Definitive diagnosis of cornealSCCwas made through histopathological examination of the surgical biopsies. Thoracic radiography and submandibular lymph node cytology revealed no evidence of metastatic neoplasia. Following healing of the corneal biopsy sites, topical 1% 5‐FUointment was applied four times daily for four consecutive days once a month, for six treatment cycles. Twenty‐three months after diagnosis, the patient remains visual and comfortable with no evidence ofSCCrecurrence. Long‐term therapy with once daily topical 1% cyclosporine solution was used to manage corneal pigmentation bilaterally.

Conclusions

The pulse‐therapy 1% 5‐FUprotocol was a successful, convenient, and cost‐effective adjunctive treatment with few adverse effects.
Nonlinear Vibrations and Chaotic Dynamics of the Laminated Composite Piezoelectric Beam
Authors: Yao Minghui , Zhang Wei , Yao Zhigang
Published: October 2014

This paper investigates the complicated dynamics behavior and the evolution law of the nonlinear vibrations of the simply supported laminated composite piezoelectric beam subjected to the axial load and the transverse load. Using the third-order shear deformation theory and the Hamilton's principle, the nonlinear governing equations of motion for the laminated composite piezoelectric beam are derived. Applying the method of multiple scales and Galerkin's approach to the partial differential governing equation, the four-dimensional averaged equation is obtained for the case of principal parametric resonance and 1:9 internal resonance. From the averaged equations obtained, numerical simulation is performed to study nonlinear vibrations of the laminated composite piezoelectric beam. The axial load, the transverse load, and the piezoelectric parameter are selected as the controlling parameters to analyze the law of complicated nonlinear dynamics of the laminated composite piezoelectric beam. Based on the results of numerical simulation, it is found that there exists the complex nonlinear phenomenon in motions of the laminated composite piezoelectric beam. In summary, numerical studies suggest that periodic motions and chaotic motions exist in nonlinear vibrations of the laminated composite piezoelectric beam. In addition, it is observed that the axial load, the transverse load and the piezoelectric parameter have significant influence on the nonlinear dynamical behavior of the beam. We can control the response of the system from chaotic motions to periodic motions by changing these parameters.
A Total Lagrangian ANCF Liquid Sloshing Approach for Multibody System Applications
Authors: Wei Cheng , Wang Liang , Shabana Ahmed A.
Published: October 2014

The objective of this investigation is to develop a total Lagrangian nonincremental liquid sloshing solution procedure based on the finite element (FE) absolute nodal coordinate formulation (ANCF). The proposed liquid sloshing modeling approach can be used to avoid the difficulties of integrating most of fluid dynamics formulations, which are based on the Eulerian approach, with multibody system (MBS) dynamics formulations, which are based on a total Lagrangian approach. The proposed total Lagrangian FE fluid dynamics formulation, which can be systematically integrated with computational MBS algorithms, differs significantly from the conventional FE or finite volume methods which are based on an Eulerian representation that employs the velocity field of a fixed control volume in the region of interest. The ANCF fluid equations are expressed in terms of displacement and gradient coordinates of material points, allowing for straightforward implementation of kinematic constraint equations and for the systematic modeling of the interaction of the fluid with the external environment or with rigid and flexible bodies. The fluid incompressibility conditions and surface traction forces are considered and derived directly from the Navier–Stokes equations. Two ANCF brick elements, one is obtained using an incomplete polynomial representation and the other is obtained from a B-spline volume representation, are used. The new approach ensures the continuity of the displacement gradients at the nodal points and allows for imposing higher degree of continuity across the element interface by applying algebraic constraint equations that can be used to eliminate dependent variables and reduce the model dimensionality. Regardless of the magnitude of the fluid displacement, the fluid has a constant mass matrix, leading to zero Coriolis and centrifugal forces. The analysis presented in this paper demonstrates the feasibility of developing an efficient nonincremental total Lagrangian approach for modeling sloshing problems in MBS system applications in which the bodies can experience large displacements including finite rotations. Several examples are presented in order to shed light on the potential of using the ANCF liquid sloshing formulation developed in this study.
Evaluation of Two Low-Constraint Toughness Test Methods in a Single Specimen
Authors: Park Dong-Yeob , Gravel Jean-Philippe , Arafin Muhammad , Liang Jie , Hari Manoj Simha C.
Published: October 2014

In previous studies, the single- and double-clip gauge methods were successfully consolidated in a single-edge notched tension (SE(T)) single specimen so that crack tip opening displacement (CTOD) values obtained from both SE(T) methods could be compared under identical test conditions. The current study investigated the effect of unloading compliance crack size equations on resistance curves obtained from both gauging methods combined in a single specimen. It was found that the unloading compliance crack size equations of Cravero and Ruggieri and the single clip gauge method predict crack sizes well within approximately 2% error in average. Two CTOD-resistance curves obtained from both gauging methods produce approximately the same results until peak loads, and thereafter the curves deviate. The results obtained from the double clip gauge method are consistently higher than those from the single clip gauge, although the difference between two resistance curves is reduced when the same unloading compliance crack size prediction procedure is used. This observation is very important within the framework of engineering critical assessment (ECA) and defect assessment procedures. An “apparent” higher resistance curve will generate larger tolerable defects thereby reducing the conservatism of an ECA analysis.
Extrinsic Tau-Coupling and the Regulation of Interceptive Reaching Under Varying Task Constraints
Authors: Smith Joanne , Grealy Madeleine , Pepping Gert-Jan
Published: October 2014

We investigated a general theory accounting for the guidance of ongoing movements in an interceptive reaching task. The aim was to assess the premise of tau-coupling that the coupling constant k, the ratio of taus (τs) of motion gaps between hand and object, reflects the kinematics of the on-going movement. The spatial and temporal constraints of the interceptive action were manipulated in three task conditions. While the time dependent counterpart of k, K(t) exhibited task effects, k itself could not distinguish between task manipulations. K(t) showed large variability during the initial acceleration phase, small variability during the rest of the movement, and task dependent changes during the final deceleration phase of interception. The findings highlight the importance of clarifying what constitutes as t-coupling.
A Comprehensive Functional Map of the Hepatitis C Virus Genome Provides a Resource for Probing Viral Proteins
Authors: Remenyi Roland , Qi Hangfei , Su Sheng-Yao , Chen Zugen , Wu Nicholas C. , Arumugaswami Vaithilingaraja , Truong Shawna , Chu Virginia , Stokelman Tamar , Lo Hung-Hao , Olson C. Anders , Wu Ting-Ting , Chen Shu-Hwa , Lin Chung-Yen , Sun Ren
Published: October 2014

Pairing high-throughput sequencing technologies with high-throughput mutagenesis enables genome-wide investigations of pathogenic organisms. Knowledge of the specific functions of protein domains encoded by the genome of the hepatitis C virus (HCV), a major human pathogen that contributes to liver disease worldwide, remains limited to insight from small-scale studies. To enhance the capabilities of HCV researchers, we have obtained a high-resolution functional map of the entire viral genome by combining transposon-based insertional mutagenesis with next-generation sequencing. We generated a library of 8,398 mutagenized HCV clones, each containing one 15-nucleotide sequence inserted at a unique genomic position. We passaged this library in hepatic cells, recovered virus pools, and simultaneously assayed the abundance of mutant viruses in each pool by next-generation sequencing. To illustrate the validity of the functional profile, we compared the genetic footprints of viral proteins with previously solved protein structures. Moreover, we show the utility of these genetic footprints in the identification of candidate regions for epitope tag insertion. In a second application, we screened the genetic footprints for phenotypes that reflected defects in later steps of the viral life cycle. We confirmed that viruses with insertions in a region of the nonstructural protein NS4B had a defect in infectivity while maintaining genome replication. Overall, our genome-wide HCV mutant library and the genetic footprints obtained by high-resolution profiling represent valuable new resources for the research community that can direct the attention of investigators toward unidentified roles of individual protein domains.

IMPORTANCE
Our insertional mutagenesis library provides a resource that illustrates the effects of relatively small insertions on local protein structure and HCV viability. We have also generated complementary resources, including a website (http://hangfei.bol.ucla.edu) and a panel of epitope-tagged mutant viruses that should enhance the research capabilities of investigators studying HCV. Researchers can now detect epitope-tagged viral proteins by established antibodies, which will allow biochemical studies of HCV proteins for which antibodies are not readily available. Furthermore, researchers can now quickly look up genotype-phenotype relationships and base further mechanistic studies on the residue-by-residue information from the functional profile. More broadly, this approach offers a general strategy for the systematic functional characterization of viruses on the genome scale.
LpxC Inhibitors as New Antibacterial Agents and Tools for Studying Regulation of Lipid A Biosynthesis in Gram-Negative Pathogens
Authors: Tomaras Andrew P. , McPherson Craig J. , Kuhn Michael , Carifa Arlene , Mullins Lisa , George David , Desbonnet Charlene , Eidem Tess M. , Montgomery Justin I. , Brown Matthew F. , Reilly Usa , Miller Alita A. , O’Donnell John P.
Published: October 2014

The problem of multidrug resistance in serious Gram-negative bacterial pathogens has escalated so severely that new cellular targets and pathways need to be exploited to avoid many of the preexisting antibiotic resistance mechanisms that are rapidly disseminating to new strains. The discovery of small-molecule inhibitors of LpxC, the enzyme responsible for the first committed step in the biosynthesis of lipid A, represents a clinically unprecedented strategy to specifically act against Gram-negative organisms such as
Pseudomonas aeruginosa
and members of the
Enterobacteriaceae
. In this report, we describe the microbiological characterization of LpxC-4, a recently disclosed inhibitor of this bacterial target, and demonstrate that its spectrum of activity extends to several of the pathogenic species that are most threatening to human health today. We also show that spontaneous generation of LpxC-4 resistance occurs at frequencies comparable to those seen with marketed antibiotics, and we provide an in-depth analysis of the mechanisms of resistance utilized by target pathogens. Interestingly, these isolates also served as tools to further our understanding of the regulation of lipid A biosynthesis and enabled the discovery that this process occurs very distinctly between
P. aeruginosa
and members of the
Enterobacteriaceae
. Finally, we demonstrate that LpxC-4 is efficacious
in vivo
against multiple strains in different models of bacterial infection and that the major first-step resistance mechanisms employed by the intended target organisms can still be effectively treated with this new inhibitor.

IMPORTANCE
New antibiotics are needed for the effective treatment of serious infections caused by Gram-negative pathogens, and the responsibility of identifying new drug candidates rests squarely on the shoulders of the infectious disease community. The limited number of validated cellular targets and approaches, along with the increasing amount of antibiotic resistance that is spreading throughout the clinical environment, has prompted us to explore the utility of inhibitors of novel targets and pathways in these resistant organisms, since preexisting target-based resistance should be negligible. Lipid A biosynthesis is an essential process for the formation of lipopolysaccharide, which is a critical component of the Gram-negative outer membrane. In this report, we describe the
in vitro
and
in vivo
characterization of novel inhibitors of LpxC, an enzyme whose activity is required for proper lipid A biosynthesis, and demonstrate that our lead compound has the requisite attributes to warrant further consideration as a novel antibiotic.
Change in prevalence of chronic kidney disease in England over time: comparison of nationally representative cross-sectional surveys from 2003 to 2010
Authors: Aitken Grant R , Roderick Paul J , Fraser Simon , Mindell Jennifer S , O'Donoghue Donal , Day Julie , Moon Graham
Published: October 2014

Objectives

To determine whether the prevalence of chronic kidney disease (CKD) in England has changed over time.

Design

Cross-sectional analysis of nationally representative Health Survey for England (HSE) random samples.

Setting

England 2003 and 2009/2010.

Survey participants

13 896 adults aged 16+ participating in HSE, adjusted for sampling and non-response, 2009/2010 surveys combined.

Main outcome measure

Change in prevalence of estimated glomerular filtration rate (eGFR) <60 mL/min/1.73 m2(as proxy for stage 3–5 CKD), from 2003 to 2009/2010 based on a single serum creatinine measure using an isotope dilution mass spectrometry traceable enzymatic assay in a single laboratory; eGFR derived using Modified Diet in Renal Disease (MDRD) and Chronic Kidney Disease Epidemiology Collaboration (CKDEPI) eGFR formulae.

Analysis

Multivariate logistic regression modelling to adjust time changes for sociodemographic and clinical factors (body mass index, hypertension, diabetes, lipids). A correction factor was applied to the 2003 HSE serum creatinine to account for a storage effect.

Results

National prevalence of low eGFR (<60) decreased within each age and gender group for both formulae except in men aged 65–74. Prevalence of obesity and diabetes increased in this period, while there was a decrease in hypertension. Adjustment for demographic and clinical factors led to a significant decrease in CKD between the surveyed periods. The fully adjusted OR for eGFR <60 mL/min/1.73 m2was 0.75 (0.61 to 0.92) comparing 2009/2010 with 2003 using the MDRD equation, and was similar using the CKDEPI equation 0.73 (0.57 to 0.93).

Conclusions

The prevalence of a low eGFR indicative of CKD in England appeared to decrease over this 7-year period, despite the rising prevalence of obesity and diabetes, two key causes of CKD. Hypertension prevalence declined and blood pressure control improved but this did not appear to explain the fall. Periodic assessment of eGFR and albuminuria in future HSEs is needed to evaluate trends in CKD.
Short-term use of remifentanil during endotracheal extubation for prophylactic analgesia in neurosurgical patients after craniotomy (SURE after Craniotomy Study): a study protocol and statistical analysis plan for a randomised controlled trial
Authors: Wu Yuan-Xing , Chen Han , Zhou Jian-Xin
Published: October 2014

Introduction

Acute pain is common during the endotracheal extubation period, and is related to complications and adverse outcomes. Patients with delayed extubation after craniotomy are vulnerable to pain and complications of extubation. However, pain control during extubation is still inadequate. Remifentanil, a new opioid with rapid onset and short duration of action, provides adequate analgesia during procedures with minimal effect of respiratory depression.

Methods and analysis

The study is a prospective, randomised, double-blinded, controlled parallel-group design. Patients with delayed extubation after intracranial surgery are screened daily. Adult patients ready for extubation are enrolled and assigned randomly to one of the two treatment study groups, labelled as the ‘Remi group’ or ‘Saline group’. Patients in the Remi group receive an intravenous bolus dose of remifentanil 0.5 μg/kg over 60 s followed by a continuous infusion 0.05 μg/kg/min for 20 min. Patients in the Saline group receive an intravenous infusion of 0.9% sodium chloride at a volume and rate equal to that of remifentanil. Pain intensity is measured by the visual analogue scale (VAS) pain score. Adverse events during drug infusion are documented and reported. Patients will be followed up until hospital discharge, death or 60 days after the trial intervention on a first come, first served basis. Details of the incidence of reintubation and reoperation within 72 h after extubation, length of stay in the intensive care unit and hospital and mortality are collected. The primary end point is the incidence of severe pain (defined as a VAS pain score more than 5 cm) during the periextubation period (defined as the period of time from immediately before extubation to 20 min after extubation).

Ethics and dissemination

The study was approved by the Institutional Review Board (IRB) of the Beijing Tiantan Hospital, Capital Medical University. The study findings will be disseminated through peer-reviewed publications and conference presentations.

Trial registration number

ClinicalTrials (NCT): ChiCTR-PRC-13003879.
Study protocol for a double blind, randomised, placebo-controlled trial of continuous subpectoral local anaesthetic infusion for pain and shoulder function following mastectomy: SUB-pectoral Local anaesthetic Infusion following MastEctomy (SUBLIME) study
Authors: Langford R , Brown I , Vickery J , Mitchell K , Pritchard C , Creanor S
Published: October 2014

Introduction

Over 16 000 mastectomies are performed in England and Wales annually. Acute postoperative pain and nausea are common. The most frequently occurring long-term complications are chronic pain (up to 50%) and reduced shoulder function (reported at 35%). Regional techniques that improve acute postoperative pain relief may reduce the incidence of these complications. This study assesses the effectiveness of a 24-hour continuous local anaesthetic in the subpectoral plane in improving postoperative pain and quality of life in patients undergoing mastectomy.

Methods and analysis

This is a randomised, double blind, placebo-controlled, two-centre, parallel group trial in women undergoing mastectomy with or without axillary involvement. One hundred and sixty participants will be randomised in a 1:1 ratio to receive either 0.25% levobupivacaine or 0.9% saline by subpectoral infusion postoperatively for 24 h. All participants will be provided with an intravenous morphine patient-controlled analgesia (PCA) system. Participants will be followed-up for 24 h in hospital and at approximately 14 days and 6 months postoperatively. Joint primary outcome measures are total morphine consumption and total pain score (captured via patient-recorded visual analogue scale (VAS) 4 hourly) during the first 24 h postoperatively. Primary statistical analysis of total pain is based on the area under the curve of pain versus time graph. Secondary outcomes include PCA attempts in first 24 h; VAS pain scores and shoulder function by goniometry at 24 h, 14 days (approximately) and 6 months; Verbal Rating Scale pain scores in first 24 h; Brief Pain Inventory and Oxford Shoulder Score at 6 months; duration of hospital stay; incidence of postoperative nausea and vomiting; cost-effectiveness.

Ethics and dissemination

The study is approved by the South West England Research Ethics Committee (12/SW/0149).

Results

will be published in a peer-reviewed journal and presented at local, national and international scientific meetings.

Trial registration

ISRCTN46621916. EudraCT 2011-005775-16.
Key characteristics of successful quality improvement curricula in physician education: a realist review
Authors: Jones Anne C , Shipman Scott A , Ogrinc Greg
Published: October 2014

Purpose

Quality improvement (QI) is a common competency that must be taught in all physician training programmes, yet, there is no clear best approach to teach this content in clinical settings. We conducted a realist systematic review of the existing literature in QI curricula within the clinical setting, highlighting examples of trainees learning QI by doing QI.

Method

Candidate theories describing successful QI curricula were articulateda priori. We searched MEDLINE (1 January 2000 to 12 March 2013), the Cochrane Library (2013) and Web of Science (15 March 2013) and reviewed references of prior systematic reviews. Inclusion criteria included study design, setting, population, interventions, clinical and educational outcomes. The data abstraction tool included categories for setting, population, intervention, outcomes and qualitative comments. Themes were iteratively developed and synthesised using realist review methodology. A methodological quality tool assessed the biases, confounders, secular trends, reporting and study quality.

Results

Among 39 studies, most were before–after design with resident physicians as the primary population. Twenty-one described clinical interventions and 18 described educational interventions with a mean intervention length of 6.58 (SD=9.16) months. Twenty-eight reported successful clinical improvements; no studies reported clinical outcomes that worsened. Characteristics of successful clinical QI curricula include attention to the interface of educational and clinical systems, careful choice of QI work for the trainees and appropriately trained local faculty.

Conclusions

This realist review identified success characteristics to guide training programmes, medical schools, faculty, trainees, accrediting organisations and funders to further develop educational and improvement resources in QI educational programmes.
Efficient application of next-generation sequencing for the diagnosis of rare genetic syndromes
Authors: Madrigal Irene , Alvarez-Mora Maria Isabel , Karlberg Olof , Rodríguez-Revenga Laia , Elurbe Dei M , Rabionet Raquel , Mur Antonio , Pie Juan , Ballesta Francisca , Sauer Sascha , Syvänen Ann-Christine , Milà Montserrat
Published: October 2014

Aims

The causes of intellectual disability, which affects 1%–3% of the general population, are highly heterogeneous and the genetic defect remains unknown in around 40% of patients. The application of next-generation sequencing is changing the nature of biomedical diagnosis. This technology has quickly become the method of choice for searching for pathogenic mutations in rare uncharacterised genetic diseases.

Methods

Whole-exome sequencing was applied to a series of families affected with intellectual disability in order to identify variants underlying disease phenotypes.

Results

We present data of three families in which we identified the disease-causing mutations and which benefited from receiving a clinical diagnosis: Cornelia de Lange, Cohen syndrome and Dent-2 disease. The genetic heterogeneity and the variability in clinical presentation of these disorders could explain why these patients are difficult to diagnose.

Conclusions

The accessibility to next-generation sequencing allows clinicians to save much time and cost in identifying the aetiology of rare diseases. The presented cases are excellent examples that demonstrate the efficacy of next-generation sequencing in rare disease diagnosis.
Recanalization of acute basilar artery occlusion improves outcomes: a meta-analysis
Authors: Kumar Gyanendra , Shahripour Reza Bavarsad , Alexandrov Andrei V
Published: October 2014

Objective

Acute basilar artery occlusion (BAO) is a catastrophe with high mortality and disability. Randomized controlled trial (RCT) data supporting/refuting reperfusion treatment are sparse in acute BAO. The study objective was to pool risk estimates for death or dependency (DoD), mortality, and intracerebral hemorrhage (ICH) from published studies on BAO.

Methods

Medline, Embase, The Cochrane Library, and clinicaltrials.gov were searched through August 2013 for studies of reperfusion therapies in acute BAO. Two independent authors selected studies and abstracted the data. Pooled risk ratios (RR) and mean weighted probabilities (MWP) were calculated using random effects model.

Results

45 studies (n=2056) met the inclusion criteria. Recanalization was associated with a lower risk of DoD (RR 0.67; number needed to treat (NNT) 3) and mortality (RR 0.49; NNT 2.5). ICH rate (MWP) with acute therapies was 13%. With recanalization, RR for DoD in those treated ≤12 h was 0.63, and for those treated >12 h, it was 0.67. With recanalization, RR for DoD in the intravenous thrombolysis (IVT) subgroup was 0.68; in the intra-arterial/endovascular therapy (IA/EVT) subgroup, DoD RR was 0.67. Recanalization resulted in mortality RR of 0.46 in those treated ≤12 h; for >12 h group, RR was 0.50. Recanalization resulted in mortality RR of 0.53 in the IVT subgroup, and RR of 0.48 in the IA/EVT subgroup. ICH rate for ≤12 h was 10%; ICH rate for >12 h was 14%. With IVT, the ICH rate was 9%, and with IA/EVT the ICH rate was 14%.

Conclusions

Recanalization of acute BAO leads to reduction in mortality by two-fold and reduction in the risk of DoD by 1.5-fold. This study underscores the equipoise in acute BAO for comparing systemic and endovascular therapies, and testing different time windows through RCTs.
Characterization of phospholipid-encapsulated gold nanoparticles: a versatile platform to study drug delivery and cellular uptake mechanisms
Authors: Wang Meijing , Petersen Nils O.
Published: October 2014

The data presented in this work aim to provide a comprehensive characterization of lipid-coated gold nanoparticles. We show that it is possible to envelop gold nanoparticles with a coating of lipids during the formation of the gold nanoparticles, that the gold in these lipid-coated gold nanoparticles is crystalline and the size and shape can be controlled by the reaction conditions (within limits), that the lipid coating corresponds to a thickness consistent with the formation of a bilayer, that the bilayer can include fluorescent probes that, while quenched, can be used to trace the fate of the gold nanoparticles in cellular systems, that their surface charge, and hence their overall stability in solution, is influenced by the lipid coating, and that while some lipid dyes may exchange among the particles, they are sufficiently stable to exchange to permit their use as tracers in cell studies. We believe this is the most comprehensive characterization of these systems to date.