bluuebunny/crossref_metadata_2025 huggingface dataset:
Simple variables predict miserable outcome after intravenous thrombolysis
Authors: Seiffge D. J. , Karagiannis A. , Strbian D. , Gensicke H. , Peters N. , Bonati L. H. , Kotisaari K. , Leppä M. , Kejda‐Scharler J. , Tränka C. , Ginsbach P. , Tatlisumak T. , Lyrer P. A. , Engelter S. T.
Published: September 2013

Background and purpose

To test the predictability of miserable outcome amongst ischaemic stroke patients receiving intravenous thrombolysis (IVT) based on a simple variables model (SVM) and to compare the model's predictive performance with that of an existing score which includes imaging and laboratory parameters (DRAGON).

Methods

The SVM consists of the parameters age, independence before stroke, normal Glasgow coma verbal score, able to lift arms and able to walk. In a derivation cohort (n = 1346) and a validation cohort (n = 638) of consecutive IVT‐treated stroke patients, the probability estimated by SVM and the observed occurrence of miserable 3‐month outcome (modified Rankin score 5–6) were compared. The performances of SVM and the DRAGON score were compared. The area under the receiver operating curve (AUC) (95% confidence interval, CI) and the bootstrapping approach were used to compare the predictive performance.

Results

The AUCs to predict miserable outcome in the derivation cohort were 0.807 (95% CI 0.774–0.838) using the SVM and 0.822 (0.790–0.850) using the DRAGON score (P = 0.3). For the validation cohort, AUCs were 0.786 (0.742–0.829) for the SVM and 0.809 (0.774–0.845) for the DRAGON score (P = 0.23). Only one patient with an SVM probability of >70% for miserable outcome in either cohort had a good outcome whilst 83% had a miserable outcome. An online SVM calculator to estimate the probability of miserable outcome for individual patients is available under http://www.unispital-basel.ch/SVM‐Tool.

Conclusion

The SVM was similar in accuracy to the DRAGON score for predicting miserable outcome after IVT. As these simple variables are available already at the pre‐hospital stage, the SVM may facilitate and accelerate pre‐hospital triage of patients at high risk for miserable outcome after IVT towards endovascular treatment.
Squaring the circle: reconciling fishing and conservation of aquatic ecosystems
Authors: Law Richard , Kolding Jeppe , Plank Michael J
Published: September 2013

Size‐at‐entry regulations in fisheries cause major disruption to aquatic ecosystems, including truncation of age‐ and size‐structures, destabilization of fish stocks, directional selection on phenotypic traits and a by‐catch of unwanted species and sizes. Here, we use simple dynamic models of size‐spectra to examine an alternative, so‐called balanced harvesting. Balanced harvesting helps in retaining the approximate power‐law size‐structure of natural ecosystems, whereas size‐at‐entry regulations do not. Balanced harvesting is less likely to destabilize steady states than size‐at‐entry regulations set close to the size at maturation. Surprisingly, our numerical results suggest that steady‐state biomass yield can be substantially increased by switching from size‐at‐entry to balanced harvesting. On the basis of these results, we argue that the goals of conservation and of greater yields seem less difficult to reconcile than have previously been thought. However, to work towards these goals require a change in our approach to fishing.
Mechanistic insights into 1‐deoxy‐d‐xylulose 5‐phosphate reductoisomerase, a key enzyme of the MEP terpenoid biosynthetic pathway
Authors: Li Heng , Tian Jie , Sun Wei , Qin Wei , Gao Wen‐Yun
Published: September 2013

The binding mode of 1‐deoxy‐d‐xylulose 5‐phosphate (DXP) to 1‐deoxy‐d‐xylulose 5‐phosphate reductoisomerase (DXR) (EC 1.1.1.267) from Escherichia coli was investigated via 18O isotope exchange experiments and determination of the kinetic parameters of the reaction. The results support a C3–C4 substrate binding mode in which DXP chelates a DXR‐bound divalent cation via its hydroxyl groups at C3 and C4. Based on this binding mode and the early results, a catalytic cycle for the conversion of DXP to 2‐methyl‐d‐erythritol 4‐phosphate mediated by DXR including a pseudo‐single molecule transition state of the retro‐aldol intermediates is proposed. Taking into account the binding mode of DXP and the catalytic cycle of DXR, the mechanistic insights of DXR are disclosed and the current discrepancies concerning the catalysis of this enzyme are interpreted within the accepted retro‐aldol/aldol sequence.
The effect of modified atmosphere packaging on the microbiological, physical, chemical and sensory characteristics of broadtail squid (Illex coindetii)
Authors: Bouletis Achilleas D. , Arvanitoyannis Ioannis S. , Hadjichristodoulou Christos , Neofitou Christos , Sakkomitrou Maria , Kolokythopoulou Foteini
Published: September 2013

The aim of this work was to evaluate the effect of various atmosphere compositions (20% CO2/80% N2 for modified atmosphere packaging (MAP) 1, 50% CO2/50% N2 for MAP 2, 70% CO2/30% N2 for MAP 3 and vacuum packaging) on the microbial (mesophiles, psychrophiles, Pseudomonas spp., Brochothrix thermosphacta and Enterobacteriaceae), physical, chemical [trimethylamine (TMA) and total volatile basic nitrogen (TVBN)] and sensorial characteristics of broadtail squid (Illex coindetii) stored for 10 days at 2 ± 1 °C. All microbial populations were severely restrained by MAP 3 with the exception of Enterobacteriaceae, which seemed to take advantage of the lack of competitive microflora and had enhanced microbial counts on MAP samples (P < 0.05). Colour attributes were better maintained on MAP‐stored samples. Drip loss was less on vacuum‐packaged squids. MAP 2 was the best atmosphere for the preservation of tissue consistency. TMA and TVBN formation was limited by high CO2 atmospheres, even though both elevated in all studied conditions. Shelf life based on sensory characteristics was determined to be 10, 8, 6, 6 and 4 days for MAP 3, MAP 2, MAP 1, vacuum and control samples, respectively.
Variations in lipid and fatty acid contents in different body parts of Black Sea whiting, Merlangius merlangus euxinus (Nordmann, 1840)
Authors: Tufan Bekir , Köse Sevim
Published: September 2013

Whiting is a commercially important fish species of the world. This study demonstrates monthly variations in lipid and fatty acid (FA) contents of muscle, liver and roes of Black Sea whiting, Merlangius merlangus euxinus. Significant changes occurred in lipid contents between months (P < 0.05) with the highest values representing in liver 33.8–64.5%. Total polyunsaturated fatty acids (PUFA) in all groups were higher than total saturated and monounsaturated FAs with significant variations between months (P < 0.05). The highest PUFA of muscles, livers and roes were 60.0, 45.9 and 50.9%, respectively. The main FA was docosahexaenoic acid (DHA) of muscle tissue and roes, while oleic acid was the major FA in livers. Although about 164–357 g in muscle tissue or 224–392 g of whiting roe are necessary to consume to cover 1 g day−1 of eicosapentaenoic acid (EPA)+DHA for a healthy diet, only as low as 5.5–10.0 g of liver would be enough to cover the same amount of daily EPA+DHA requirement. The results indicated that whiting livers constitute a rich and underexploited source of polyunsatured FAs. Furthermore, the results may aid further research on the nutritional studies, the physiology and stock management of whiting species.
Isolation, purification and characterisation of polygalacturonase from ripened banana (Musa acuminata cv. Kadali)
Authors: Gayathri Thulasy , Nair Ashalatha S.
Published: September 2013

Bananas are the most important fruit crop in the world. Short shelf life of the fruit is the major limiting factor in international trade, and this is due to the softening of the pulp during ripening. Fruit softening is an important aspect of ripening process in fleshy fruits and is caused by the cumulative action of a group of cell wall‐modifying enzymes. Polygalacturonase (PG) is the key enzyme involved in the fruit softening process in banana, and this study reports the isolation, purification and characterisation of polygalacturonase enzyme from ripened fruits of a delayed ripened banana cultivar found specifically in Kerala (Musa acuminata cv. Kadali). PG was purified by ammonium sulphate fractionation followed by DEAE cellulose ion exchange chromatography and gel filtration using Sephadex G 100. The purified protein showed two subunits on SDS‐PAGE and a single band on native PAGE. Enzyme showed maximum activity at pH 3.5 and 40 °C. Fe3+ enhanced the activity more, while Mg2+ and Ca2+ slightly stimulated the activity of purified enzyme. Km value for substrate polygalacturonic acid was 0.06%.
Effect of blanching and drying treatments on quality of bamboo shoot slices
Authors: Zheng Jiong , Zhang Fusheng , Song Jiaxin , Lin Mao , Kan Jianquan
Published: September 2013

The enzymes activity, texture, pectin, microstructure and colour change in bamboo shoots (Dendrocalamus latiflorus) were examined to evaluate the effect of blanching and drying treatments on quality of bamboo shoot slices (BSS). Peroxidase (POD), phenylalanine ammonia‐lyase and polyphenol oxidase in BSS will become completely inactive at 95 °C for 6‐, 9‐ and 12‐min blanching treatments, respectively. The hardness and protopectin (PP) content decreased after the blanching treatment. Meanwhile, the content of water‐soluble pectin increased initially and then decreased gradually during the blanching. After the blanching treatment, the L* and a* of BSS decreased, while b* increased, ΔE > 2. The hardness of rehydrated BSS decreased by 43.57% after freeze drying (FD), which, however, decreased more after hot air drying (HAD). The pectin of rehydrated BSS was also determined to explain the hardness change in BSS. The BSS acquired similar microstructure of fresh samples after FD, while microstructure of BSS was severely damaged after HAD. The colour of BSS changes significantly after drying treatment. However, FD can maintain better colour of BSS than HAD. The results may provide a reference for industrial production of BSS.
Structural, functional, thermal and rheological properties of nixtamalised and extruded blue maize (Zea maysL.) flour with different calcium sources
Authors: Sánchez‐Madrigal Miguel Ángel , Meléndez‐Pizarro Carmen O. , Martínez‐Bustos Fernando , Ruiz‐Gutiérrez Martha G. , Quintero‐Ramos Armando , Márquez‐Meléndez Rubén , Lardizábal‐Gutiérrez Daniel , Campos‐Venegas Karla
Published: September 2013

The effects of different concentrations of calcium hydroxide [Ca(OH)2] and calcium lactate [C6H10O6Ca] on the functional and physical properties of extruded (EF) and nixtamalised (NF) blue maize flours were evaluated. Calcium source and concentration showed no significant effects on theEFexpansion index. The water absorption index (WAI) ofEFdecreased as the concentrations of both calcium sources increased, andNFwith C6H10O6Ca had the lowestWAI. The thermal and pasting properties ofNFs were higher than those ofEF.NFwith C6H10O6Ca showed the highest final viscosity (FinV), indicating less damage to the starch granules, and this was correlated with microscopic analysis. In contrast, the FinV ofEFs was significantly affected by calcium source and concentration. Extrusion with 0.3% and nixtamalisation at 2.95% of C6H10O6Ca yielded highWAIvalue and the best rheological properties in maize flour, respectively. These results suggest the use of C6H10O6Ca in extrusion or nixtamalisation to produce blue maize flours for tortilla or snacks with antioxidants.
Plasma levels of high‐sensitive C‐reactive protein do not correlate with inflammatory activity in carotid atherosclerotic plaques
Authors: Grufman H. , Gonçalves I. , Edsfeldt A. , Nitulescu M. , Persson A. , Nilsson M. , Nilsson J.
Published: September 2013

Background

It is well established that subjects with moderately elevated plasma levels of C‐reactive protein (CRP) have an increased risk of development of cardiovascular events. As atherosclerosis is a disease characterized by chronic arterial inflammation, it is possible that moderate increases in CRP level reflect the presence of plaque inflammation. To investigate this possibility, we compared plasma levels of hsCRP the day before carotid endarterectomy with the degree of inflammation in the excised plaque tissue.

Methods

Luminex immunoassays were used to determine the levels of IL‐6, IL‐10, monocyte chemoattractant protein‐1 and tumour necrosis factor‐α (TNF‐α) in plasma and in homogenized plaque tissue from 160 endarterectomy specimens. Plaque sections were stained with antibodies against CD68 to determine the plaque macrophage content.

Results

Plasma high‐sensitivity (hs)CRP levels were significantly correlated with plasma IL‐6 and TNF‐α. However, there were no significant associations between plasma hsCRP concentration and plaque cytokine levels or macrophage contents.

Conclusions

The present findings strongly argue against hsCRP as a marker of plaque inflammation. Hence, it is more likely that elevated hsCRP is a sign of a subclinical systemic inflammation and this in turn may contribute to progression of cardiovascular disease.
Putting incertae sedis taxa in their place: a proposal for ten new families and three new genera in Sphaeropleales (Chlorophyceae, Chlorophyta)
Authors: Fučíková Karolina , Lewis Paul O. , Lewis Louise A.
Published: September 2013

Best known for aquatic colonial algae such as Hydrodictyon, Pediastrum, or Scenedesmus, the order Sphaeropleales also contains numerous coccoid taxa from aquatic and terrestrial habitats. Recent findings indicate that coccoid lineages in this order are very diverse genetically and may be the prevalent form, although their diversity is often hidden morphologically. This study characterizes coccoid algae recently discovered from desert soil crusts that share morphological and ecological features with the genera Bracteacoccus, Pseudomuriella, and Chromochloris. Analyses of a multi‐gene data set that includes members from all sphaeroplealean families are used to examine the monophyly of these morphologically similar taxa, which are shown instead to be phylogenetically distinct and very divergent. We propose new generic names for these lineages: Bracteamorpha, Rotundella, and Tumidella. In addition, we propose an updated family‐level taxonomy within Sphaeropleales that includes ten new families of coccoid algae to accommodate the newly presented genera and many incertae sedis taxa in the order: Bracteamorphaceae, Chromochloridaceae, Dictyococcaceae, Dictyochloridaceae, Mychonastaceae, Pseudomuriellaceae, Rotundellaceae, Schizochlamydaceae, Schroederiaceae, and Tumidellaceae.
Analytic versus systemic group therapy for women with a history of child sexual abuse: 1‐Year follow‐up of a randomized controlled trial
Authors: Elkjaer Henriette , Kristensen Ellids , Mortensen Erik L. , Poulsen Stig , Lau Marianne
Published: September 2013

Objective

This randomized prospective study examines durability of improvement in general symptomatology, psychosocial functioning and interpersonal problems, and compares the long‐term efficacy of analytic and systemic group psychotherapy in women 1 year after completion of treatment for childhood sexual abuse.

Design and Methods

Women (n = 106) randomly assigned to analytic or systemic psychotherapy completed the Symptom Checklist‐90‐R, Global Assessment of Functioning, Global Life Quality, Registration Chart Questionnaire, and Flashback Registration at pre‐treatment, post‐treatment, and at a 1‐year follow‐up.

Results

Post‐treatment gains were significant for both treatment modalities on all measures, but significantly larger after systemic therapy. Significant treatment response was maintained 1‐year post‐treatment, but different trajectories were observed: 1 year after treatment completion, improvements for analytic therapy were maintained, whereas they decreased after systemic therapy, resulting in no statistically significant difference in gains between the groups at the 1‐year follow‐up. Despite maintaining significant gains, more than half of the patients remained above cut‐off for caseness concerning general symptomatology at post‐treatment and at 1‐year follow‐up.

Conclusion

The findings stress the importance of long‐term follow‐up data in effect studies. Different trajectories were associated with the two treatments, but improvement in the two treatment groups did not differ significantly at the 1‐year follow‐up. Implications of the difference in trajectories for treatment planning are discussed.

Practitioner points

Both analytic and systemic group therapy proved efficient in improving general symptomatology, psychosocial functioning, and interpersonal problems in women with a history of CSA and gains were maintained at a 1‐year follow‐up.

Despite maintaining statistically significant gains at the 1‐year follow‐up, 54% of the patients remained above the cut‐off for caseness with respect to general symptomatology, which may indicate a need for further treatment.

Different pre‐post follow‐up treatment trajectories were observed between the two treatment modalities. Thus, while systemic group therapy showed a significantly better outcome immediately after termination, gains in the systemic treatment group decreased during follow‐up, while gains were maintained during follow‐up in analytic group therapy.
High‐frequency ultrasound imaging of tattoo reactions with histopathology as a comparative method. Introduction of preoperative ultrasound diagnostics as a guide to therapeutic intervention
Authors: Hutton Carlsen K. , Tolstrup J. , Serup J.
Published: September 2013

Background and Aim

Tattoo adverse reactions requiring diagnostic evaluation and treatment are becoming more common. The aim of this study was to assess tattoo reactions by 20‐MHz ultrasonography referenced to histopathology as a comparative method.

Materials and Methods

A total of 73 individuals with clinical adverse reactions in their tattoos were studied. Punch biopsies for reference histology were available from 58 patients. The Dermascan C® of Cortex Technology, Denmark, was employed. Total skin thickness and echo density of the echolucent band in the outer dermis were measured. Biopsy served for diagnosis and for determination of the level of cellular infiltration in the dermis.

Results

In every tattoo reaction studied, the skin affected was found thicker compared with regional control of the same individual (mean difference 0.73 mm). A prominent echolucent band of mean thickness 0.89 mm was demonstrated, primarily located in the very outer dermis but propagating to deeper dermal layers parallel to increasing severity of reactions. The thickness of the echolucent band correlated with the thickness of cellular infiltration determined by microscopic examination, R = 0.6412 (P < 0.001). Special diagnoses such as granulomatous and pseudolymphomatous tattoo reactions showed no distinct characteristics by ultrasound, but mainly displayed themselves by their advanced inflammatory component.

Discussion and Conclusion

It is demonstrated for the first time that ultrasound, with histopathology as the comparative method, can quantify the severity of tattoo reactions and non‐invasively diagnose the depth of the inflammatory process in the dermis elicited by the microparticulate tattoo pigment, which itself is too minute to be imaged by ultrasound. Preoperative 20‐MHz ultrasound scanning is introduced as a potentially useful method to guide therapeutic interventions by surgery and lasers.
Crossing Industry Borders: German Manufacturers as Services Exporters
Authors: Kelle Markus
Published: September 2013

Manufacturing firms increasingly engage in service trade activities. Microlevel data show that German manufacturers accounted for nearly 30 billion euros of service exports in 2005. I have found that particular construction, engineering and R&D services are exported. The machinery industries, motor vehicles and chemicals producers dominate the overall pattern. The types of services exported vary strongly across industries. Service exports of advertising, data processing, management and R&D services are found to likely support foreign affiliates of firms. However, these headquarter services are only infrequently observable. Much more important are construction and engineering services exported by machinery firms. These might represent installation and maintenance services complementing exported machinery. R&D services exports generally represent the transfer of knowledge and technology, which is found to be particularly relevant for motor vehicle producers. Beyond the support of foreign production of firms, R&D services might be relevant also in R&D cooperations of firms or when firms have partnerships with foreign suppliers or buyers of intermediate products.
Thermotoga lettingae Can Salvage Cobinamide To Synthesize Vitamin B12
Authors: Butzin Nicholas C. , Secinaro Michael A. , Swithers Kristen S. , Gogarten J. Peter , Noll Kenneth M.
Published: September 2013

We recently reported that theThermotogalesacquired the ability to synthesize vitamin B12by acquisition of genes from two distantly related lineages,ArchaeaandFirmicutes(K. S. Swithers et al., Genome Biol. Evol. 4:730–739, 2012). Ancestral state reconstruction suggested that the cobinamide salvage gene cluster was present in theThermotogales' most recent common ancestor. We also predicted thatThermotoga lettingaecould not synthesize B12de novobut could use the cobinamide salvage pathway to synthesize B12. In this study, these hypotheses were tested, and we found thatTt. lettingaedid not synthesize B12de novobut salvaged cobinamide. The growth rate ofTt. lettingaeincreased with the addition of B12or cobinamide to its medium. It synthesized B12when the medium was supplemented with cobinamide, and no B12was detected in cells grown on cobinamide-deficient medium. Upstream of the cobinamide salvage genes is a putative B12riboswitch. In other organisms, B12riboswitches allow for higher transcriptional activity in the absence of B12. WhenTt. lettingaewas grown with no B12, the salvage genes were upregulated compared to cells grown with B12or cobinamide. Another gene cluster with a putative B12riboswitch upstream is thebtuFCDABC transporter, and it showed a transcription pattern similar to that of the cobinamide salvage genes. The BtuF proteins from species that can and cannot salvage cobinamides were shownin vitroto bind both B12and cobinamide. These results suggest thatThermotogalesspecies can use the BtuFCD transporter to import both B12and cobinamide, even if they cannot salvage cobinamide.
Thermal Inactivation of Desiccation-Adapted Salmonella spp. in Aged Chicken Litter
Authors: Chen Zhao , Diao Junshu , Dharmasena Muthu , Ionita Claudia , Jiang Xiuping , Rieck James
Published: September 2013

Thermal inactivation of desiccation-adapted
Salmonella
spp. in aged chicken litter was investigated in comparison with that in a nonadapted control to examine potential cross-tolerance of desiccation-adapted cells to heat treatment. A mixture of four
Salmonella
serovars was inoculated into the finished compost with 20, 30, 40, and 50% moisture contents for a 24-h desiccation adaptation. Afterwards, the compost with desiccation-adapted cells was inoculated into the aged chicken litter with the same moisture content for heat treatments at 70, 75, 80, 85, and 150°C. Recovery media were used to allow heat-injured cells to resuscitate. A 5-log reduction in the number of the desiccation-adapted cells in aged chicken litter with a 20% moisture content required >6, >6, ∼4 to 5, and ∼3 to 4 h of exposure at 70, 75, 80, and 85°C, respectively. As a comparison, a 5-log reduction in the number of nonadapted control cells in the same chicken litter was achieved within ∼1.5 to 2, ∼1 to 1.5, ∼0.5 to 1, and <0.5 h at 70, 75, 80, and 85°C, respectively. The exposure time required to obtain a 5-log reduction in the number of desiccation-adapted cells gradually became shorter as temperature and moisture content were increased. At 150°C, desiccation-adapted
Salmonella
cells survived for 50 min in chicken litter with a 20% moisture content, whereas control cells were detectable by enrichment for only 10 min. Our results demonstrated that the thermal resistance of
Salmonella
in aged chicken litter was increased significantly when the cells were adapted to desiccation. This study also validated the effectiveness of thermal processing being used for producing chicken litter free of
Salmonella
contamination.
BopA Does Not Have a Major Role in the Adhesion of Bifidobacterium bifidum to Intestinal Epithelial Cells, Extracellular Matrix Proteins, and Mucus
Authors: Kainulainen Veera , Reunanen Justus , Hiippala Kaisa , Guglielmetti Simone , Vesterlund Satu , Palva Airi , Satokari Reetta
Published: September 2013

The ability of bifidobacteria to adhere to the intestine of the human host is considered to be important for efficient colonization and achieving probiotic effects.
Bifidobacterium bifidum
strains DSM20456 and MIMBb75 adhere well to the human intestinal cell lines Caco-2 and HT-29. The surface lipoprotein BopA was previously described to be involved in mediating adherence of
B. bifidum
to epithelial cells, but thioacylated, purified BopA inhibited the adhesion of
B. bifidum
to epithelial cells in competitive adhesion assays only at very high concentrations, indicating an unspecific effect. In this study, the role of BopA in the adhesion of
B. bifidum
was readdressed. The gene encoding BopA was cloned and expressed without its lipobox and hydrophobic signal peptide in
Escherichia coli
, and an antiserum against the recombinant BopA was produced. The antiserum was used to demonstrate the abundant localization of BopA on the cell surface of
B. bifidum
. However, blocking of
B. bifidum
BopA with specific antiserum did not reduce adhesion of bacteria to epithelial cell lines, arguing that BopA is not an adhesin. Also, adhesion of
B. bifidum
to human colonic mucin and fibronectin was found to be BopA independent. The recombinant BopA bound only moderately to human epithelial cells and colonic mucus, and it failed to bind to fibronectin. Thus, our results contrast the earlier findings on the major role of BopA in adhesion, indicating that the strong adhesion of
B. bifidum
to epithelial cell lines is BopA independent.
Lineage and Genogroup-Defining Single Nucleotide Polymorphisms of Escherichia coli O157:H7
Authors: Jung Woo Kyung , Bono James L. , Clawson Michael L. , Leopold Shana R. , Shringi Smriti , Besser Thomas E.
Published: September 2013

Escherichia coliO157:H7 is a zoonotic human pathogen for which cattle are an important reservoir host. Using both previously published and new sequencing data, a 48-locus single nucleotide polymorphism (SNP)-based typing panel was developed that redundantly identified 11 genogroups that span six of the eight lineages recently described forE. coliO157:H7 (J. L. Bono, T. P. Smith, J. E. Keen, G. P. Harhay, T. G. McDaneld, R. E. Mandrell, W. K. Jung, T. E. Besser, P. Gerner-Smidt, M. Bielaszewska, H. Karch, M. L. Clawson, Mol. Biol. Evol. 29:2047–2062, 2012) and additionally defined subgroups within four of those lineages. This assay was applied to 530 isolates from human and bovine sources. The SNP-based lineage groups were concordant with previously identifiedE. coliO157:H7 genotypes identified by other methods and were strongly associated with carriage of specific Stx genes. Two SNP lineages (Ia and Vb) were disproportionately represented among cattle isolates, and three others (IIa, Ib, and IIb) were disproportionately represented among human clinical isolates. This 48-plex SNP assay efficiently and economically identifies biologically relevant lineages withinE. coliO157:H7.
Rapid Sample Processing for Detection of Food-Borne Pathogens via Cross-Flow Microfiltration
Authors: Li Xuan , Ximenes Eduardo , Amalaradjou Mary Anne Roshni , Vibbert Hunter B. , Foster Kirk , Jones Jim , Liu Xingya , Bhunia Arun K. , Ladisch Michael R.
Published: September 2013

This paper reports an approach to enable rapid concentration and recovery of bacterial cells from aqueous chicken homogenates as a preanalytical step of detection. This approach includes biochemical pretreatment and prefiltration of food samples and development of an automated cell concentration instrument based on cross-flow microfiltration. A polysulfone hollow-fiber membrane module having a nominal pore size of 0.2 μm constitutes the core of the cell concentration instrument. The aqueous chicken homogenate samples were circulated within the cross-flow system achieving 500- to 1,000-fold concentration of inoculated
Salmonella enterica
serovar Enteritidis and naturally occurring microbiota with 70% recovery of viable cells as determined by plate counting and quantitative PCR (qPCR) within 35 to 45 min. These steps enabled 10 CFU/ml microorganisms in chicken homogenates or 10
2
CFU/g chicken to be quantified. Cleaning and sterilizing the instrument and membrane module by stepwise hydraulic and chemical cleaning (sodium hydroxide and ethanol) enabled reuse of the membrane 15 times before replacement. This approach begins to address the critical need for the food industry for detecting food pathogens within 6 h or less.
Restricted Substrate Specificity for the Geranylgeranyltransferase-I Enzyme in Cryptococcus neoformans: Implications for Virulence
Authors: Selvig Kyla , Ballou Elizabeth R. , Nichols Connie B. , Alspaugh J. Andrew
Published: September 2013

Proper cellular localization is required for the function of many proteins. The CaaX prenyltransferases (where CaaX indicates a cysteine followed by two aliphatic amino acids and a variable amino acid) direct the subcellular localization of a large group of proteins by catalyzing the attachment of hydrophobic isoprenoid moieties onto C-terminal CaaX motifs, thus facilitating membrane association. This group of enzymes includes farnesyltransferase (Ftase) and geranylgeranyltransferase-I (Ggtase-1). Classically, the variable (X) amino acid determines whether a protein will be an Ftase or Ggtase-I substrate, with Ggtase-I substrates often containing CaaL motifs. In this study, we identify the gene encoding the β subunit of Ggtase-I (
CDC43
) and demonstrate that Ggtase-mediated activity is not essential. However,
Cryptococcus neoformans CDC43
is important for thermotolerance, morphogenesis, and virulence. We find that Ggtase-I function is required for full membrane localization of Rho10 and the two Cdc42 paralogs (Cdc42 and Cdc420). Interestingly, the related Rac and Ras proteins are not mislocalized in the
cdc43
Δ mutant even though they contain similar CaaL motifs. Additionally, the membrane localization of each of these GTPases is dependent on the prenylation of the CaaX cysteine. These results indicate that
C. neoformans
CaaX prenyltransferases may recognize their substrates in a unique manner from existing models of prenyltransferase specificity. It also suggests that the
C. neoformans
Ftase, which has been shown to be more important for
C. neoformans
proliferation and viability, may be the primary prenyltransferase for proteins that are typically geranylgeranylated in other species.
Identification and Characterization of Non-Cellulose-Producing Mutants of Gluconacetobacter hansenii Generated by Tn
5
Transposon Mutagenesis
Authors: Deng Ying , Nagachar Nivedita , Xiao Chaowen , Tien Ming , Kao Teh-hui
Published: September 2013

The
acs
operon of
Gluconacetobacter
is thought to encode AcsA, AcsB, AcsC, and AcsD proteins that constitute the cellulose synthase complex, required for the synthesis and secretion of crystalline cellulose microfibrils. A few other genes have been shown to be involved in this process, but their precise role is unclear. We report here the use of Tn
5
transposon insertion mutagenesis to identify and characterize six non-cellulose-producing (Cel
−
) mutants of
Gluconacetobacter hansenii
ATCC 23769. The genes disrupted were
acsA
,
acsC
,

ccp
Ax

(encoding cellulose-complementing protein [the subscript “
Ax
” indicates genes from organisms formerly classified as
Acetobacter xylinum
]),
dgc1
(encoding guanylate dicyclase), and
crp-fnr
(encoding a cyclic AMP receptor protein/fumarate nitrate reductase transcriptional regulator). Protein blot analysis revealed that (i) AcsB and AcsC were absent in the
acsA
mutant, (ii) the levels of AcsB and AcsC were significantly reduced in the

ccp
Ax

mutant, and (iii) the level of AcsD was not affected in any of the Cel
−
mutants. Promoter analysis showed that the
acs
operon does not include
acsD
, unlike the organization of the
acs
operon of several strains of closely related
Gluconacetobacter xylinus
. Complementation experiments confirmed that the gene disrupted in each Cel
−
mutant was responsible for the phenotype. Quantitative real-time PCR and protein blotting results suggest that the transcription of

bgl
Ax

(encoding β-glucosidase and located immediately downstream from
acsD
) was strongly dependent on Crp/Fnr. A

bgl
Ax

knockout mutant, generated via homologous recombination, produced only ∼16% of the wild-type cellulose level. Since the
crp-fnr
mutant did not produce any cellulose, Crp/Fnr may regulate the expression of other gene(s) involved in cellulose biosynthesis.
Two DHH Subfamily 1 Proteins in Streptococcus pneumoniae Possess Cyclic Di-AMP Phosphodiesterase Activity and Affect Bacterial Growth and Virulence
Authors: Bai Yinlan , Yang Jun , Eisele Leslie E. , Underwood Adam J. , Koestler Benjamin J. , Waters Christopher M. , Metzger Dennis W. , Bai Guangchun
Published: September 2013

Cyclic di-AMP (c-di-AMP) and cyclic di-GMP (c-di-GMP) are signaling molecules that play important roles in bacterial biology and pathogenesis. However, these nucleotides have not been explored inStreptococcus pneumoniae, an important bacterial pathogen. In this study, we characterized the c-di-AMP-associated genes ofS. pneumoniae. The results showed that SPD_1392 (DacA) is a diadenylate cyclase that converts ATP to c-di-AMP. Both SPD_2032 (Pde1) and SPD_1153 (Pde2), which belong to the DHH subfamily 1 proteins, displayed c-di-AMP phosphodiesterase activity. Pde1 cleaved c-di-AMP into phosphoadenylyl adenosine (pApA), whereas Pde2 directly hydrolyzed c-di-AMP into AMP. Additionally, Pde2, but not Pde1, degraded pApA into AMP. Our results also demonstrated that both Pde1 and Pde2 played roles in bacterial growth, resistance to UV treatment, and virulence in a mouse pneumonia model. These results indicate that c-di-AMP homeostasis is essential for pneumococcal biology and disease.
The Clostridium perfringens Germinant Receptor Protein GerKC Is Located in the Spore Inner Membrane and Is Crucial for Spore Germination
Authors: Banawas Saeed , Paredes-Sabja Daniel , Korza George , Li Yunfeng , Hao Bing , Setlow Peter , Sarker Mahfuzur R.
Published: September 2013

The Gram-positive, anaerobic, spore-forming bacterium
Clostridium perfringens
causes a variety of diseases in both humans and animals, and spore germination is thought to be the first stage of
C. perfringens
infection. Previous studies have indicated that the germinant receptor (GR) proteins encoded by the bicistronic
gerKA-gerKC
operon as well as the proteins encoded by the
gerKB
and
gerAA
genes are required for normal germination of
C. perfringens
spores. We now report the individual role of these GR proteins by analyzing the germination of strains carrying mutations in
gerKA
,
gerKC
, or both
gerKB
and
gerAA
. Western blot analysis was also used to determine the location and numbers of GerKC proteins in spores. Conclusions from this work include the following: (i)
gerKC
mutant spores germinate extremely poorly with KCl,
l
-asparagine, a mixture of asparagine and KCl, or NaP
i
; (ii)
gerKC
spores germinate significantly more slowly than wild-type and other GR mutant spores with a 1:1 chelate of Ca
2+
and dipicolinic acid and very slightly more slowly with dodecylamine; (iii) the germination defects in
gerKC
spores are largely restored by expressing the wild-type
gerKA-gerKC
operon in
trans
; (iv) GerKC is required for the spores' viability, almost certainly because of the
gerKC
spores' poor germination; and (v) GerKC is located in the spores' inner membrane, with ∼250 molecules/spore. Collectively, these results indicate that GerKC is the main GR protein required for nutrient and nonnutrient germination of spores of
C. perfringens
food-poisoning isolates.
THE VALUE OF ADMITTING PATIENTS FOLLOWING A POSITIVE MYOGLOBIN USING A RAPID POINT OF CARE TRIPLE CARDIAC PANEL TEST
Authors: Coates J , Walker C
Published: September 2013

Objectives & Background

Admissions to the Emergency Department with chest pain constitute a significant proportion of the work-load. In England it represents 6% of all Emergency Department (ED) attendances. These attendances translate to accounting for approximately 25% of acute medical admissions. One method of excluding myocardial infarction is the use of a rapid point of care Triple Cardiac Marker test. This allows testing at time point 0 and 90 minutes and negates the need for a delayed troponin. One of the markers, myoglobin, has a high sensitivity but low specificity. If there is a 25% rise in myoglobin between two tests then it is considered a positive result. The patient then requires a 12-hour troponin. Locally, there was concern over the value of including myoglobin in the triple test as it was felt that it lead to inappropriate admissions.

Methods

This was a retrospective case review. The Triage© Cardiac Panel machine from Alere records patient numbers and test results. Patient numbers and results were extracted over a continuous 8 month period. The Emergency Department notes were then interrogated and this was combined with data from the hospital pathology system, looking specifically for patients that were admitted on the basis of positive myoglobin result. Hospital notes were requested if points needed to be clarified.

Results

A total of 636 paired tests were analysed. 103 (16.3%) of patients had a positive myoglobin rise. The range of myoglobin rises can be seen in Figure 1. 100 notes were available, of which only 56 were admitted for a 12 hour troponin. 54 underwent 12 hour troponin testing and three were positive (5.6%). There appeared to be no correlation between the % rise in myoglobin and the rise in troponin. 44 patients were discharged after achieving a positive triple test result. 35 of these were felt to be inappropriate. 16 sets of notes reported mentioned somewhere that the triple test was “negative.” The use of the chest pain proforma did not seem to improve the performance of the doctors–Figure 2.

Abstract 011 Figure 1

Range of Myoglobin Rises.

Abstract 011 Figure 2

Effect of completion of Chest Pain Protocol on patient disposition.

Conclusion

The use of myoglobin in the triple test does appear to be appropriate for the local population. There is a significant short-coming in the application of the triple test that is putting patients at risk of an adverse outcome. The current chest pain proforma as it stands does not appear to prevent inappropriate discharges.
INTRODUCING CHECKLISTS FOR SEDATION AND INTUBATION INTO THE EMERGENCY DEPARTMENT; A CHALLENGE WORTH RISING TO?
Authors: Galloway R I , Swann F
Published: September 2013

Objectives & Background

The Emergency Department (ED) is a fertile ground for medical error. In numerous other areas checklists (and the associated cultural changes) have been brought in to reduce error. Following the National Audit Project into airway complications in 2011, guidelines were produced recommending the use of a checklist prior to rapid sequence induction (RSI) along with capnography in the ED. Similarly, College of Emergency Medicine (CEM) guidelines recommended ‘checks’ prior to sedation and capnography use. The project objectives were firstly to review the national use of checklists (and capnography). Secondly, within one trust which has already implemented the checklists, to ascertain if they were used in reality. Finally, the study looked into attitudes and how barriers to implementation could be overcome.

Methods

A telephone survey of all English Emergency Departments was conducted to determine the use of checklists (and capnography) for intubation and sedation. Within Brighton and Sussex University (BSUH) NHS trust, a retrospective notes audit was conducted to determine checklist use. Finally, staff involved in patient care were interviewed regarding their experiences of checklists.

Results

138 out of 178 EDs agreed to take part in the survey. 48.6% and 58% of the EDs reported using a RSI and sedation checklist, respectively. An association (p<0.0003) between checklist use and capnography in sedation was identified. The audit data from BSUH NHS trust showed checklists were used in 21.5% of RSIs and 57.8% of sedations. Interviews showed that barriers included problems of implementation across multiple specialties and overcoming staff's pre-conceived attitudes towards the checklists.

Conclusion

The use of checklists within Emergency Medicine remains limited. Their uptake within a trust that has implemented them has been slow. Cultural resistance within the workplace remains a barrier that needs to be overcome.
SEE AND TREAT FOR MAJOR TRAUMA? AN EVALUATION OF THE USE OF A TWO-TIERED TRAUMA TEAM ACTIVATION SYSTEM IN A UK MAJOR TRAUMA CENTRE
Authors: Jenkins P E , Rogers J , Kehoe A , Smith J
Published: September 2013

Objectives & Background

Appropriate activation of multi-disciplinary trauma teams improves outcome for severely injured patients, but can disrupt normal service in the rest of the hospital. Derriford Hospital uses a two-tiered trauma team activation system. The emergency department (ED) trauma team is activated in response to a significant traumatic mechanism; the hospital trauma team is activated when this mechanism co-exists with physiological abnormality or specific anatomical injury. The aim of this study was to compare characteristics, process measures and outcomes between patients treated by ED or hospital trauma teams to evaluate the approach in a UK setting and to estimate any cost savings involved. Figure 1 outlines the composition and activation criteria of the teams.

Abstract 014 Figure 1

(a) ED trauma team activation and (b) Hospital trauma team activation.

Methods

A retrospective database review was performed, using the TARN database and the local source trauma database. All patients who activated a trauma team between 1 Apr 12 to 30 Sep 12 were included. Patients were categorised according to the type of trauma team activated to receive them. Data included time to primary survey X-rays, time to CT, time to intubation, numbers discharged from ED, ICU admission, injury severity score (ISS), and mortality. Cost savings associated with activating an ED rather than hospital response were extrapolated from previously described methodology.

Results

During the study period 456 patients activated a trauma team, 358 ED and 98 hospital (see Figure 2). Patients seen by the ED team were significantly less likely to have severe injury or require hospital admission, intubation, emergency operation or blood transfusion. Differences in time taken to key investigations were statistically but not clinically significant. The cost saving of avoiding hospital team activation for those patients not requiring specialist care was estimated at £180,663 per annum.

Abstract 014 Figure 2

Statistics comparing Hospital and ED calls.

Conclusion

A two-tiered trauma team activation system is an efficient, safe and cost-effective way of dealing with trauma patients presenting to a Major Trauma Centre in the UK.
WHEN DO ED TRAINEES GET ASSESSMENTS DONE?
Authors: Townend W
Published: September 2013

Objectives & Background

Formative assessments support learning. Ideally then, such trainee/ trainer interaction would occur throughout the training year and include cases that are stretching. In this way areas for trainee development could be identified and a training plan refined and executed. Such an approach would give time to revisit themes with further assessments towards the end of the year.

Methods

The CEM e-portfolio was interrogated and anonymised data retrieved. Records for ‘Formative mini-cex’ were collated for 3rd August 2011 to 4th August 2012. The date and outcome were recorded for each assessment

Results

2022 ‘Formative mini-cex’ records were found. Of these, 94 (5%) were completed in the first 2 months, and 787 (39%) in the 2 months prior to ARCP. The monthly completion of these assessments was August 24 (1.4%), September 70 (3.5%), October 119 (6%), November 128 (6.3%), December 103 (5%), January 245 (12%), February 118 (6%), March 135 (6.7%), April 181 (8.9%), May 343 (17%), June 444 (22%), July 101 (5%). Each episode included an opportunity to evaluate eight domains of practice. 85% of evaluations across all domains were of excellent practice. 14% of evaluations included recommendations that the trainee ‘should consider…’ and 1% that the trainee ‘must consider…'. Written narrative feedback was recorded in 1874 (93%) of assessments. 1346 (67%) were assessed by consultants, 177 (9%) by SAS doctors and 495 (25%) by ST4-6 trainees.

Conclusion

There is a high rate of excellence recorded is assessments, and around half of assessments are performed at the end of the academic year. There were more assessments performed in the month of ARCP (June) than in the first 5 months of the academic year. Future iterations of the assessment schedule will include means of ensuring trainee/trainer interaction is scheduled throughout the year and includes a case mix that stretches the trainee with trainer support.
ACUTE ALLERGY AND ANAPHYLAXIS IN THE EMERGENCY DEPARTMENT–A REVIEW OF 1 YEAR'S PATIENT PRESENTATIONS
Authors: Noble R , Friedlaender G , Cuthbertson B , Gray A
Published: September 2013

Objectives & Background

There is limited epidemiological data for allergy Emergency Department (ED) presentations. Following recent launch of NICE guidelines and World Allergy Organisation (WAO) severity descriptions we investigated the epidemiology, management and outcomes of allergy patients presenting to a single ED.

Methods

A retrospective review of consecutive presentations to a single tertiary ED (assessing 112,000 adults annually) between 28/02/2011 and 29/02/2012 was conducted. Key-word search of ED PAS records identified a suitable cohort. WAO criteria were used to define severity of allergic presentation.

Results

405 patients were identified (268 acute allergy patients, 137 anaphylaxis). Median age 33 (range 13 to 92) years. Male to female ratio was 0.6:1. 28 patients met criteria for life-threatening reactions. 259 (64%) patients self-presented and 141 (34.8%) presented via ambulance. 6 (21.4%) patients with life-threatening anaphylaxis self-presented. Precipitants were identified in 235 (58%) patients. 89 (21.9%) patients had a known allergy to the precipitating agent, including 38 (27.7%) anaphylaxis patients. 33 (12.8%) acute allergy and 65 (47.4%) anaphylaxis patients received pre-hospital adrenaline (48 from ambulance service, 49 self-administered). In the ED, 103 (75.2%) anaphylaxis patients received H1 antagonists, 86 (62.8%) received oral or IV steroids, 24 (17.5%) received IM adrenaline and 23 (16.8%) received salbutamol. Only 63 (15.5%; n=405) were admitted to hospital including 3 anaphylaxis patients to critical care. Of those admitted; 57 (90.5%) received adrenaline pre-hospital or in the ED. Overall 21 (5%) patients re-presented within 72 hours of discharge; 3 of these patients had been admitted. An epipen was prescribed on discharge to 82 (20.2%) patients overall and 44 (10.9%) were referred to allergy services. 34 (54%) admitted patients were not referred to specialists and 36 (57.1%) did not receive an epipen.

Conclusion

NICE guidelines were not consistently followed but this did not seem to result in measurable short terms complications. A significant number of patients had a known precipitant.