bluuebunny/crossref_metadata_2025 huggingface dataset:
Site-Directed Mutations in the Lanthipeptide Mutacin 1140
Authors: Chen Shaorong , Wilson-Stanford Shawanda , Cromwell William , Hillman Jeffrey D. , Guerrero Adam , Allen Charlotte A. , Sorg Joseph A. , Smith Leif
Published: April 2013

The oral bacterium
Streptococcus mutans
, strain JH1140, produces the antibiotic mutacin 1140. Mutacin 1140 belongs to a group of antibiotics called lanthipeptides. More specifically, mutacin 1140 is related to the epidermin type A(I) lanthipeptides. Mutagenesis experiments of this group of lanthipeptides have been primarily restricted to the posttranslationally modified
meso
-lanthionine and 3-methyllanthionine residues. Site-directed mutagenesis of the core peptide of mutacin 1140 was performed using the suicide vector pVA891. Substitutions of the N-terminal residue, the charged residue in the hinge region, and residues in ring A and intertwined rings C and D were investigated. A truncation and insertion of residues in ring A and intertwined rings C and D were also performed to determine whether or not they would alter the antimicrobial activity of the producing strain. Bioassays revealed that five of 14 mutants studied had improved antimicrobial activity against the indicator strain
Micrococcus luteus
ATCC 10240. MICs against
Streptococcus mutans
UA159,
Streptococcus pneumoniae
ATCC 27336,
Staphylococcus aureus
ATCC 25923,
Clostridium difficile
UK1, and
Micrococcus luteus
ATCC 10240 were determined for three mutacin 1140 variants that had the most significant increases in bioactivity in the
M. luteus
bioassay. This mutagenesis study of the epidermin group of lanthipeptides shows that antimicrobial activity can be significantly improved.
Dechlorination of Chloral Hydrate Is Influenced by the Biofilm Adhesin Protein LapA in Pseudomonas putida LF54
Authors: Zhang Wanjun , Huhe  , Pan Yuanbai , Toyofuku Masanori , Nomura Nobuhiko , Nakajima Toshiaki , Uchiyama Hiroo
Published: April 2013

LapA is the largest surface adhesion protein of
Pseudomonas putida
that initiates biofilm formation. Here, by using transposon insertion mutagenesis and a conditional
lapA
mutant, we demonstrate for the first time that LapA influences chloral hydrate (CH) dechlorination in
P. putida
LF54.
Development of a Reverse Transcription-Quantitative PCR System for Detection and Genotyping of Aichi Viruses in Clinical and Environmental Samples
Authors: Kitajima Masaaki , Hata Akihiko , Yamashita Teruo , Haramoto Eiji , Minagawa Hiroko , Katayama Hiroyuki
Published: April 2013

Aichi viruses (AiVs) have been proposed as a causative agent of human gastroenteritis potentially transmitted by fecal-oral routes through contaminated food or water. In the present study, we developed a TaqMan minor groove binder (MGB)-based reverse transcription-quantitative PCR (RT-qPCR) system that is able to quantify AiVs and differentiate between genotypes A and B. This system consists of two assays, an AiV universal assay utilizing a universal primer pair and a universal probe and a duplex genotype-specific assay utilizing the same primer pair and two genotype-specific probes. The primers and probes were designed based on multiple alignments of the 21 available AiV genome sequences containing the capsid gene. Using a 10-fold dilution of plasmid DNA containing the target sequences, it was confirmed that both assays allow detection and quantification of AiVs with a quantitative range of 1.0 × 10
1
to 1.0 × 10
7
copies/reaction, and the genotype-specific assay reacts specifically to each genotype. To validate the newly developed assays, 30 clinical stool specimens were subsequently examined with the assays, and the AiV RNA loads were determined to be 1.4 × 10
4
to 6.6 × 10
9
copies/g stool. We also examined 12 influent and 12 effluent wastewater samples collected monthly for a 1-year period to validate the applicability of the assays for detection of AiVs in environmental samples. The AiV RNA concentrations in influent and effluent wastewater were determined to be up to 2.2 × 10
7
and 1.8 × 10
4
copies/liter, respectively. Our RT-qPCR system is useful for routine diagnosis of AiVs in clinical stool specimens and environmental samples.
In Vivo
Assessment of Growth and Virulence Gene Expression during Commensal and Pathogenic Lifestyles of
luxABCDE
-Tagged Enterococcus faecalis Strains in Murine Gastrointestinal and Intravenous Infection Models
Authors: La Rosa Sabina Leanti , Casey Pat G. , Hill Colin , Diep Dzung B. , Nes Ingolf F. , Brede Dag A.
Published: April 2013

Cytolysin and gelatinase are prominent pathogenicity determinants associated with highly virulent
Enterococcus faecalis
strains. In an effort to explore the expression profiles of these virulence traits
in vivo
, we have employed
E. faecalis
variants expressing the
luxABCDE
cassette under the control of either the P

16S

, cytolysin, or gelatinase promoter for infections of
Galleria mellonella
caterpillars and mice. Systemic infection of
G. mellonella
with bioluminescence-tagged
E. faecalis
MMH594 revealed temporal regulation of both gelatinase and cytolysin promoters and demonstrated that these traits were induced in response to the host environment. Gavage of mice pretreated perorally with antibiotics resulted in efficient colonization of the murine gastrointestinal tract (GIT) in a strain-dependent manner, where the commensal baby isolate EF62 was more persistent than the nosocomial isolate MMH594. A highly significant correlation (
R
2
> 0.94) was found between bioluminescence and the CFU counts in mouse fecal samples. Both strains showed similar preferences for growth and persistence in the ileum, cecum, and colon. Cytolysin expression was uniform in these compartments of the intestinal lumen. In spite of high numbers (10
9
CFU/g of intestinal matter) in the ileum, cecum, and colon, no evidence of translocation or systemic infection could be observed. In the murine intravenous infection model, cytolysin expression was readily detected in the liver, kidneys, and bladder. At 72 h postinfection, the highest bacterial loads were found in the liver, kidneys, and spleen, with organ-specific expression levels of cytolysin ∼400- and ∼900-fold higher in the spleen and heart, respectively, than in the liver and kidneys. Taken together, this system based on the bioluminescence imaging technology is established as a new, powerful method to monitor the differential regulation of
E. faecalis
virulence determinants and to study the spatiotemporal course of infection in living animals in real time.
Biofilm Formation by Psychrobacter arcticus and the Role of a Large Adhesin in Attachment to Surfaces
Authors: Hinsa-Leasure Shannon M. , Koid Cassandra , Tiedje James M. , Schultzhaus Janna N.
Published: April 2013

Psychrobacter arcticus
strain 273-4, an isolate from a Siberian permafrost core, is capable of forming biofilms when grown in minimal medium under laboratory conditions. Biofilms form at 4 to 22°C when acetate is supplied as the lone carbon source and with 1 to 7% sea salt.
P. arcticus
is also capable of colonizing quartz sand. Transposon mutagenesis identified a gene important for biofilm formation by
P. arcticus
. Four transposon mutants were mapped to a 20.1-kbp gene, which is predicted to encode a protein of 6,715 amino acids (Psyc_1601). We refer to this open reading frame as
cat1
, for cold attachment gene 1. The
cat1
mutants are unable to form biofilms at levels equivalent to that of the wild type, and there is no impact on the planktonic growth characteristics of the strains, indicating a specific role in biofilm formation. Through time course studies of the static microtiter plate assay, we determined that
cat1
mutants are unable to form biofilms equivalent to that of the wild type under all conditions tested. In flow cell experiments,
cat1
mutants initially are unable to attach to the surface. Over time, however, they form microcolonies, an architecture very different from that produced by wild-type biofilms. Our results demonstrate that Cat1 is involved in the initial stages of bacterial attachment to surfaces.
Characterization of
Tt
ALV2, an Essential Charged Repeat Motif Protein of the Tetrahymena thermophila Membrane Skeleton
Authors: El-Haddad Houda , Przyborski Jude M. , Kraft Lesleigh G. K. , McFadden Geoffrey I. , Waller Ross F. , Gould Sven B.
Published: April 2013

Alveolins are a recently described class of proteins common to all members of the superphylum Alveolata that are characterized by conserved charged repeat motifs (CRMs) but whose exact function remains unknown. We have analyzed the smaller of the two alveolins of
Tetrahymena thermophila
,
Tt
ALV2. The protein localizes to dispersed, broken patches arranged between the rows of the longitudinal microtubules. Macronuclear knockdown of
Ttalv2
leads to multinuclear cells with no apparent cell polarity and randomly occurring cell protrusions, either by interrupting pellicle integrity or by disturbing cytokinesis. Correct association of
Tt
ALV2 with the alveoli or the pellicle is complex and depends on both the termini as well as the charged repeat motifs of the protein. Proteins containing similar CRMs are a dominant part of the ciliate membrane cytoskeleton, suggesting that these motifs may play a more general role in mediating membrane attachment and/or cytoskeletal association. To better understand their integration into the cytoskeleton, we localized a range of CRM-based fusion proteins, which suggested there is an inherent tendency for proteins with CRMs to be located in the peripheral cytoskeleton, some nucleating as filaments at the basal bodies. Even a synthetic protein, mimicking the charge and repeat pattern of these proteins, directed a reporter protein to a variety of peripheral cytoskeletal structures in
Tetrahymena
. These motifs might provide a blueprint for membrane and cytoskeleton affiliation in the complex pellicles of Alveolata.
A Central Role for Ras1 in Morphogenesis of the Basidiomycete Schizophyllum commune
Authors: Knabe Nicole , Jung Elke-Martina , Freihorst Daniela , Hennicke Florian , Horton J. Stephen , Kothe Erika
Published: April 2013

Fungi have been used as model systems to define general processes in eukaryotes, for example, the one gene-one enzyme hypothesis, as well as to study polar growth or pathogenesis. Here, we show a central role for the regulator protein Ras in a mushroom-forming, filamentous basidiomycete linking growth, pheromone signaling, sexual development, and meiosis to different signal transduction pathways.
ras1
and Ras-specific
gap1
mutants were generated and used to modify the intracellular activation state of the Ras module. Transformants containing constitutive
ras1
alleles (
ras1
G12V
and
ras1
Q61L
), as well as their compatible mating interactions, did show strong phenotypes for growth (associated with Cdc42 signaling) and mating (associated with mitogen-activated protein kinase signaling). Normal fruiting bodies with abnormal spores exhibiting a reduced germination rate were produced by outcrossing of these mutant strains. Homozygous Δ
gap1
primordia, expected to experience increased Ras signaling, showed overlapping phenotypes with a block in basidium development and meiosis. Investigation of cyclic AMP (cAMP)-dependent protein kinase A indicated that constitutively active
ras1
, as well as Δ
gap1
mutant strains, exhibit a strong increase in Tpk activity. Ras1-dependent, cAMP-mediated signal transduction is, in addition to the known signaling pathways, involved in fruiting body formation in
Schizophyllum commune
. To integrate these analyses of Ras signaling, microarray studies were performed. Mutant strains containing constitutively active Ras1, deletion of RasGap1, or constitutively active Cdc42 were characterized and compared. At the transcriptome level, specific regulation highlighting the phenotypic differences of the mutants is clearly visible.
Mutations in Escherichia coli ExbB Transmembrane Domains Identify Scaffolding and Signal Transduction Functions and Exclude Participation in a Proton Pathway
Authors: Baker Kristin R. , Postle Kathleen
Published: April 2013

The TonB system couples cytoplasmic membrane proton motive force (pmf) to active transport of diverse nutrients across the outer membrane. Current data suggest that cytoplasmic membrane proteins ExbB and ExbD harness pmf energy. Transmembrane domain (TMD) interactions between TonB and ExbD allow the ExbD C terminus to modulate conformational rearrangements of the periplasmic TonB C terminus
in vivo
. These conformational changes somehow allow energization of high-affinity TonB-gated transporters by direct interaction with TonB. While ExbB is essential for energy transduction, its role is not well understood. ExbB has N-terminus-out, C-terminus-in topology with three TMDs. TMDs 1 and 2 are punctuated by a cytoplasmic loop, with the C-terminal tail also occupying the cytoplasm. We tested the hypothesis that ExbB TMD residues play roles in proton translocation. Reassessment of TMD boundaries based on hydrophobic character and residue conservation among distantly related ExbB proteins brought earlier widely divergent predictions into congruence. All TMD residues with potentially function-specific side chains (Lys, Cys, Ser, Thr, Tyr, Glu, and Asn) and residues with probable structure-specific side chains (Trp, Gly, and Pro) were substituted with Ala and evaluated in multiple assays. While all three TMDs were essential, they had different roles: TMD1 was a region through which ExbB interacted with the TonB TMD. TMD2 and TMD3, the most conserved among the ExbB/TolQ/MotA/PomA family, played roles in signal transduction between cytoplasm and periplasm and the transition from ExbB homodimers to homotetramers. Consideration of combined data excludes ExbB TMD residues from direct participation in a proton pathway.
Periodic Reversals in Paenibacillus dendritiformis Swarming
Authors: Be'er Avraham , Strain Shinji K. , Hernández Roberto A. , Ben-Jacob Eshel , Florin E.-L.
Published: April 2013

Bacterial swarming is a type of motility characterized by a rapid and collective migration of bacteria on surfaces. Most swarming species form densely packed dynamic clusters in the form of whirls and jets, in which hundreds of rod-shaped rigid cells move in circular and straight patterns, respectively. Recent studies have suggested that short-range steric interactions may dominate hydrodynamic interactions and that geometrical factors, such as a cell's aspect ratio, play an important role in bacterial swarming. Typically, the aspect ratio for most swarming species is only up to 5, and a detailed understanding of the role of much larger aspect ratios remains an open challenge. Here we study the dynamics of
Paenibacillus dendritiformis C
morphotype, a very long, hyperflagellated, straight (rigid), rod-shaped bacterium with an aspect ratio of ∼20. We find that instead of swarming in whirls and jets as observed in most species, including the shorter
T
morphotype of
P. dendritiformis
, the
C
morphotype moves in densely packed straight but thin long lines. Within these lines, all bacteria show periodic reversals, with a typical reversal time of 20 s, which is independent of their neighbors, the initial nutrient level, agar rigidity, surfactant addition, humidity level, temperature, nutrient chemotaxis, oxygen level, illumination intensity or gradient, and cell length. The evolutionary advantage of this unique back-and-forth surface translocation remains unclear.
Core-Gene-Encoded Peptide Regulating Virulence-Associated Traits in Streptococcus mutans
Authors: Kim Jeong Nam , Stanhope Michael J. , Burne Robert A.
Published: April 2013

Recently, high-coverage genome sequence of 57 isolates of
Streptococcus mutans
, the primary etiological agent of human dental caries, was completed. The SMU.1147 gene, encoding a 61-amino-acid (61-aa) peptide, was present in all sequenced strains of
S. mutans
but absent in all bacteria in current databases. Reverse transcription-PCR revealed that SMU.1147 is cotranscribed with
scnK
and
scnR
, which encode the histidine kinase and response regulator, respectively, of a two-component system (TCS). The C terminus of the SMU.1147 gene product was tagged with a FLAG epitope and shown to be expressed in
S. mutans
by Western blotting with an anti-FLAG antibody. A nonpolar mutant of SMU.1147 formed less biofilm in glucose-containing medium and grew slower than did the wild-type strain under aerobic and anaerobic conditions, at low pH, or in the presence of H
2
O
2
. Mutation of SMU.1147 dramatically reduced genetic competence and expression of
comX
and
comY
, compared to
S. mutans
UA159. The competence defect of the SMU.1147 mutant could not be overcome by addition of
sigX
-inducing peptide (XIP) in defined medium or by competence-stimulating peptide (CSP) in complex medium. Complementation with SMU.1147 on a plasmid restored all phenotypes. Interestingly, mutants lacking either one of the TCS components and a mutant lacking all three genes behaved like the wild-type strain for all phenotypes mentioned above, but all mutant strains grew slower than UA159 in medium supplemented with 0.3 M NaCl. Thus, the SMU.1147-encoded peptide affects virulence-related traits and dominantly controls quorum-sensing pathways for development of genetic competence in
S. mutans
.
Paracoccus denitrificans PD1222 Utilizes Hypotaurine via Transamination Followed by Spontaneous Desulfination To Yield Acetaldehyde and, Finally, Acetate for Growth
Authors: Felux Ann-Katrin , Denger Karin , Weiss Michael , Cook Alasdair M. , Schleheck David
Published: April 2013

Hypotaurine (HT; 2-aminoethane-sulfinate) is known to be utilized by bacteria as a sole source of carbon, nitrogen, and energy for growth, as is taurine (2-aminoethane-sulfonate); however, the corresponding HT degradation pathway has remained undefined. Genome-sequenced
Paracoccus denitrificans
PD1222 utilized HT (and taurine) quantitatively for heterotrophic growth and released the HT sulfur as sulfite (and sulfate) and HT nitrogen as ammonium. Enzyme assays with cell extracts suggested that an HT-inducible HT:pyruvate aminotransferase (Hpa) catalyzes the deamination of HT in an initial reaction step. Partial purification of the Hpa activity and peptide fingerprinting-mass spectrometry (PF-MS) identified the Hpa candidate gene; it encoded an archetypal taurine:pyruvate aminotransferase (Tpa). The same gene product was identified via differential PAGE and PF-MS, as was the gene of a strongly HT-inducible aldehyde dehydrogenase (Adh). Both genes were overexpressed in
Escherichia coli
. The overexpressed, purified Hpa/Tpa showed HT:pyruvate-aminotransferase activity. Alanine, acetaldehyde, and sulfite were identified as the reaction products but not sulfinoacetaldehyde; the reaction of Hpa/Tpa with taurine yielded sulfoacetaldehyde, which is stable. The overexpressed, purified Adh oxidized the acetaldehyde generated during the Hpa reaction to acetate in an NAD
+
-dependent reaction. Based on these results, the following degradation pathway for HT in strain PD1222 can be depicted. The identified aminotransferase converts HT to sulfinoacetaldehyde, which desulfinates spontaneously to acetaldehyde and sulfite; the inducible aldehyde dehydrogenase oxidizes acetaldehyde to yield acetate, which is metabolized, and sulfite, which is excreted.
FrnE, a Cadmium-Inducible Protein in Deinococcus radiodurans, Is Characterized as a Disulfide Isomerase Chaperone
In Vitro
and for Its Role in Oxidative Stress Tolerance
In Vivo
Authors: Khairnar Nivedita P. , Joe Min-Ho , Misra H. S. , Lim Sang-Yong , Kim Dong-Ho
Published: April 2013

Deinococcus radiodurans
R1 exposed to a lethal dose of cadmium shows differential expression of a large number of genes, including
frnE
(dr
frnE
) and some of those involved in DNA repair and oxidative stress tolerance. The dr
frnE
::
nptII
mutant of
D. radiodurans
showed growth similar to that of the wild type, but its tolerance to 10 mM cadmium and 10 mM diamide decreased by ∼15- and ∼3-fold, respectively. These cells also showed nearly 6 times less resistance to gamma radiation at 12 kGy and ∼2-fold-higher sensitivity to 40 mM hydrogen peroxide than the wild type. In
trans
expression of drFrnE increased cytotoxicity of dithiothreitol (DTT) in the
dsbA
mutant of
Escherichia coli
. Recombinant drFrnE showed disulfide isomerase activity and could maintain insulin in its reduced form in the presence of DTT. While an equimolar ratio of wild-type protein could protect malate dehydrogenase completely from thermal denaturation at 42°C, the C22S mutant of drFrnE provided reduced protection to malate dehydrogenase from thermal inactivation. These results suggested that drFrnE is a protein disulfide isomerase
in vitro
and has a role in oxidative stress tolerance of
D. radiodurans
possibly by protecting the damaged cellular proteins from inactivation.
Molecular diagnostics and the public health management of legionellosis
Authors: Yates Tom A , Bruin Jacob P , Harrison Timothy G , Mannes Trish
Published: April 2013

In 2009–2010, we investigated fourlegionellacases notified over an 8-month period in two adjacent villages in South East England. Molecular techniques enabled us to conclude that three of the cases had distinct infections. The absence of an adequate respiratory sample in one case necessitated epidemiological investigations to exclude a potential common environmental source of further infections. One of the cases had spent a part of their incubation period in a country in South East Asia. DNA-sequence-based typing of their isolate showed it to be of theLegionella pneumophilaserogroup 1 (LP1) DNA-sequence type (ST) 481. Intriguingly, the only other two ST 481 isolates in the European Working Group for Legionella Infections database were among Dutch travellers to the same country in 2003 and 2006. This case makes clear the value of molecular diagnostics and the importance of obtaining adequate clinical specimens. The potential future uses for typing data are discussed.
Caecal lymphangioma: a rare cause of gastrointestinal blood loss
Authors: Rai P , Rao R N , Chakraborthy S B D
Published: April 2013

Lymphangioma is the malformation of the lymphatic system. Lymphangioma is a benign tumour and most colonic lymphangiomas do not cause symptoms and do not require treatment. Resection is required in patients with bleeding or intussusceptions. We report a case of intestinal lymphangiomatosis in a 31-year-old man who presented with recurrent melena and anaemia, which were diagnosed endoscopically and treated with surgical resection.
Mixed collecting duct and renal cell carcinoma presenting with spinal cord compression
Authors: Hennessey Derek B , Thomas Arun Z , Lynch Thomas H
Published: April 2013

Collecting duct carcinoma (CDC) is a rare renal malignancy thought to develop from the collecting duct epithelium of the kidney. CDC tends to have a more aggressive clinical course than conventional renal cell carcinoma (RCC), with early metastases. The occurrence of a mixed CDC and conventional RCC is infrequently reported in the literature. We report the first case of a metastatic mixed CDC and RCC presenting as back pain in a young adult. In addition we discuss the epidemiology of and current adjuvant therapies for CDC.
Treatment imprudence leading to missed tooth fragment
Authors: Barua Pranamee , Chaudhary Seema , Kaur Harsimran , Mallikarjuna Rachappa
Published: April 2013

Traumatic dental injuries (TDI) represent one of the most common oral health problems in children and adolescents. Dental trauma requires a special consideration when it accompanies soft tissue lacerations. Tooth fragments occasionally penetrate into soft tissues and may cause severe complications. This article describes the case of a 12-year-old girl with a fractured tooth fragment embedded in the lower lip for 4 months, which went unnoticed at her primary health centre. This report highlights the importance of proper radiographic diagnosis along with clinical examination after trauma in order to prevent any future complications.
20-year-old injury: an unusual cause of stroke
Authors: Lim V W L , Flynn C A , O'Connor M , Lyons D
Published: April 2013

A 62-year-old man presented with a 2-day history of headache, nausea and dizziness on a background of recently diagnosed stage 2c prostate carcinoma and controlled hypertension. Imaging confirmed bilateral cerebellar strokes; on further investigations, he was found to have undiagnosed subclavian steal syndrome following severe injury to his left arm more than 20 years earlier.
Syncope associated with water pipe smoking
Authors: Karaca Yunus , Eryigit Umut , Aksut Nurhak , Turkmen Suha
Published: April 2013

The water pipe (narghile) in particular is widely used in the Arabian Peninsula and the Turkish world, and has also recently become an increasingly popular way of consuming tobacco in Europe. Contrary to popular belief, it contains more tar, carbon monoxide (CO) and toxic gases than cigarettes. This report describes a patient presenting to the emergency department with syncope as a result of water pipe use, with tests revealing toxically high CO levels.
An open-label cohort study of the improvement of quality of life and pain in de novo cervical dystonia patients after injections with 500 U botulinum toxin A (Dysport)
Authors: Hefter H , Benecke R , Erbguth F , Jost W , Reichel G , Wissel J
Published: April 2013

Objectives

It remains to be determined whether the benefits of botulinum toxin type A (BoNT-A) on cervical dystonia (CD) motor symptoms extend to improvements in patient's quality of life (QoL). This analysis of a large, multicentre study was conducted with the aim of investigating changes in QoL and functioning among de novo patients receiving 500 U BoNT-A (abobotulinumtoxinA; Dysport) for the treatment of the two most frequent forms of CD, predominantly torticollis and laterocollis.

Design

A prospective, open-label study of Dysport (500 U; Ipsen Biopharm Ltd) administered according to a defined intramuscular injection algorithm.

Setting

German and Austrian outpatient clinics.

Participants

516 male and female patients (aged ≥18 years) with de novo CD. The majority of patients had torticollis (78.1%). 35 patients had concomitant depression (MedDRA-defined).

Main outcome measures

Change from baseline to weeks 4 and 12 in Craniocervical Dystonia Questionnaire (CDQ-24) total and subscale scores, patient diary items (‘day-to-day capacities and activities’, ‘pain’ and ‘duration of pain’) and global assessment of pain.

Results

Significant improvements were observed in CDQ-24 total and subscale scores at week 4 and were sustained up to week 12 (p<0.001). Changes in CDQ-24 scores did not significantly differ between the torticollis and laterocollis groups or between patients with or without depression. There were also significant reductions in patient diary item scores for activities of daily living, pain and pain duration at weeks 4 and 12 (p<0.001). Pain relief (less or no pain) was reported by 66% and 74.1% of patients at weeks 4 and 12, respectively. Changes in pain parameters demonstrated a positive relationship with change in Tsui score.

Conclusions

After standardised open-label treatment with Dysport 500 U, improvements in QoL and pain intensity up to 12 weeks in patients with CD were observed.
Mediation of smoking-associated postoperative mortality by perioperative complications in veterans undergoing elective surgery: data from Veterans Affairs Surgical Quality Improvement Program (VASQIP)—a cohort study
Authors: Singh Jasvinder A , Hawn Mary , Campagna Elizabeth J , Henderson William G , Richman Joshua , Houston Thomas K
Published: April 2013

Objective

To assess the mediation of smoking-associated postoperative mortality by postoperative complications.

Design

Observational cohort study.

Setting

Using data from the Veterans Affairs (VA) Surgical Quality Improvement Programme, a quality assurance programme for major surgical procedures in the VA healthcare system, we assessed the association of current smoking at the time of the surgery with 6-month and 1-year mortality.

Primary and secondary outcome measures

Using mediation analyses, we calculated the relative contribution of each smoking-associated complication to smoking-associated postoperative mortality, both unadjusted and adjusted for age, race/ethnicity, work relative value unit of the operation, surgeon specialty, American Society of Anesthesiologists class and year of surgery. Smoking-associated complications included surgical site infection (SSI), cardiovascular complications (myocardial infarction, cardiac arrest and/or stroke) and pulmonary complications (pneumonia, failure to wean and/or reintubation).

Results

There were 186 632 never smokers and 135 741 current smokers. The association of smoking and mortality was mediated by smoking-related complications with varying effects. In unadjusted analyses, the proportions of mediation of smoking to 6-month mortality explained by the complications were as follows: SSIs 22%, cardiovascular complications 12% and pulmonary complications 89%. In adjusted analyses, the per cents mediated by each complication were as follows: SSIs 2%, cardiovascular complications 4% and pulmonary complications 22%. In adjusted analyses for 1-year mortality, respective per cents mediated were 2%, 3% and 16%.

Conclusions

Pulmonary complications, followed by cardiovascular complications and SSIs were mediators of smoking-associated 6-month and 1-year mortality. Interventions targeting smoking cessation and prevention and early treatment of pulmonary complications has the likelihood of reducing postoperative mortality after elective surgery.
The effect of experience on the sensitivity and specificity of the whispered voice test: a diagnostic accuracy study
Authors: McShefferty David , Whitmer William M , Swan Iain R C , Akeroyd Michael A
Published: April 2013

Objectives

To determine the sensitivity and specificity of the whispered voice test (WVT) in detecting hearing loss when administered by practitioners with different levels of experience.

Design

Diagnostic accuracy study of WVT, through acoustic analysis of whispers of experienced and inexperienced practitioners (experiment 1) and behavioural validation of these recordings (experiment 2).

Setting

Research institute with a pool of patients sourced from local clinics in the Greater Glasgow area.

Participants

22 people had their whispers recorded and analysed in experiment 1; 4 older experienced (OE), 4 older inexperienced (OI) and 14 younger inexperienced (YI). In experiment 2, 73 people (112 individual ears) took part in a digit recognition task using 2 OE and 2 YI whisperers from experiment 1.

Main outcome measures

Average level (dB sound pressure level) across frequency, average level across all utterances (dB A) and within/across-digit deviation (dB A) for experiment 1. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of WVT for experiment 2.

Results

In experiment 1, OE whisperers were 8–10 dB more intense than inexperienced whisperers across all whispered utterances. Variability was low and comparable regardless of age or experience. In experiment 2, at an optimum threshold of 40 dB HL, sensitivity and specificity were 63% (95% CI of 58% to 68%) and 93% (92% to 94%), respectively, for OE whisperers. PPV was 56% (51% to 61%), NPV was 95% (94% to 96%). For YI whisperers at an optimum threshold of 29 dB HL, sensitivity and specificity were 80% (78% to 82%) and 52% (50% to 55%), respectively. PPV was 65% (63% to 67%) and NPV was 70% (67% to 72%).

Conclusions

WVT is an effective screening test, providing the level of the whisperer is considered when setting the test's hearing-loss criterion. Possible implications are voice measurement while training for inexperienced whisperers.
Patterns in the recording of vital signs and early warning scores: compliance with a clinical escalation protocol
Authors: Hands Chris , Reid Eleanor , Meredith Paul , Smith Gary B , Prytherch David R , Schmidt Paul E , Featherstone Peter I
Published: April 2013

Background

The recognition of patient deterioration depends largely on identifying abnormal vital signs, yet little is known about the daily pattern of vital signs measurement and charting.

Methods

We compared the pattern of vital signs and VitalPAC Early Warning Score (ViEWS) data collected from admissions to all adult inpatient areas (except high care areas, such as critical care units) of a NHS district general hospital from 1 May 2010 to 30 April 2011, to the hospital's clinical escalation protocol. Main outcome measures were hourly and daily patterns of vital signs and ViEWS value documentation; numbers of vital signs in the periods 08:00–11:59 and 20:00–23:59 with subsequent vital signs recorded in the following 6 h; and time to next observation (TTNO) for vital signs recorded in the periods 08:00–11:59 and 20:00–23:59.

Results

950 043 vital sign datasets were recorded. The daily pattern of observation documentation was not uniform; there were large morning and evening peaks, and lower night-time documentation. The pattern was identical on all days. 23.84% of vital sign datasets with ViEWS ≥ 9 were measured at night compared with 10.12–19.97% for other ViEWS values. 47.42% of patients with ViEWS=7–8 and 31.22% of those with ViEWS ≥ 9 in the period 20:00–23:59 did not have vital signs recorded in the following 6 h. TTNO decreased with increasing ViEWS value, but less than expected by the monitoring protocol.

Conclusions

There was only partial adherence to the vital signs monitoring protocol. Sicker patients appear more likely to have vital signs measured overnight, but even their observations were often not followed by timely repeat assessments. The observed pattern of monitoring may reflect the impact of competing clinical priorities.
Accuracy of prognosis prediction by PPI in hospice inpatients with cancer: a multi-centre prospective study
Authors: Subramaniam Sivakumar , Thorns Andrew , Ridout Martin , Thirukkumaran Thiru , Osborne Thomas Richard
Published: April 2013

The Palliative Prognostic Index (PPI) is a prognostication tool for palliative care patients based on clinical indices developed in Japan and further validated by one study in the UK. The aim of this study was to test its prediction accuracy in a large inpatient hospice sample. The admitting doctor in three inpatient hospices calculated the PPI score on admission. Two hundred and sixty-two patients were included in this study. Based on the PPI score, three subgroups were identified. Group 1 corresponded to patients with PPI ≤4 and the median survival of 53 days (95% CI 40 to 80 days). Group 2 corresponded to those with PPI >4 and ≤6 and the median survival 15 days (95% CI 12 to 26 days) and Group 3 corresponded to patients with PPI >6 and the median survival of 5 days (95% CI 3 to 7 days). In this study, PPI was able to identify patients’ likelihood of dying within 3 weeks with a sensitivity of 64% and specificity of 83%. It was able to identify a 6-week survival chance with a sensitivity of 62% and specificity of 86%. A one-unit increase in PPI score was estimated to increase the hazard for death by a factor of 1.33 (95% CI 1.26 to 1.40), based on fitting a stratified Cox proportional hazards model. The authors conclude that PPI can be used to predict prognosis for patients with advanced cancer.
Barriers to research in palliative care in France
Authors: Rhondali Wadih , Berthiller Julien , Hui David , Yennu Sriram , Lafumas Veronique , Ledoux Mathilde , Strasser Florian , Filbet Marilène
Published: April 2013

Purpose

Palliative care (PC) needs expansion of its research capacity to improve the quality of care. This is particularly true for France that contributed less than 2% of all European PC research publications. We conducted a survey to assess the barriers French healthcare professionals (HCPs) involved in PC research had to face.

Methods

We surveyed all 420 PC departments registered in the French National Association for Palliative Care (SFAP) database using a previously used questionnaire. We documented the ethical limitations, time constraints, financial resources, patient issues and methodological issues related to PC research.

Results

We obtained 382 responses. Ninety-two (24.1%) HCPs were involved in a research project during the last 5 years. In univariate results, predictors of being involved in PC research were men (p=0.004), physician (p=0.022), working at a university hospital (p<0.001). There was a trend towards working in a PC unit (p=0.052). The main barriers to participating in PC research were lack of time (80.1%) and patient issues (47.4%). Lack of methodological support (33.0%) and financial limitations (30.4%) were also reported as major barriers.

Conclusions

There is a consensus that PC research and publication in the English language for peer-reviewed journals must be expanded in France but at this stage, clinical teams still lack specific funding and appropriate support. More research is needed to confirm our results and to determine the best ways to develop PC research capacity in France.
Evaluation of the DAVROS (Development And Validation of Risk-adjusted Outcomes for Systems of emergency care) risk-adjustment model as a quality indicator for healthcare
Authors: Wilson Richard , Goodacre Steve W , Klingbajl Marcin , Kelly Anne-Maree , Rainer Tim , Coats Tim , Holloway Vikki , Townend Will , Crane Steve
Published: April 2013

Background and objective

Risk-adjusted mortality rates can be used as a quality indicator if it is assumed that the discrepancy between predicted and actual mortality can be attributed to the quality of healthcare (ie, the model has attributional validity). The Development And Validation of Risk-adjusted Outcomes for Systems of emergency care (DAVROS) model predicts 7-day mortality in emergency medical admissions. We aimed to test this assumption by evaluating the attributional validity of the DAVROS risk-adjustment model.

Methods

We selected cases that had the greatest discrepancy between observed mortality and predicted probability of mortality from seven hospitals involved in validation of the DAVROS risk-adjustment model. Reviewers at each hospital assessed hospital records to determine whether the discrepancy between predicted and actual mortality could be explained by the healthcare provided.

Results

We received 232/280 (83%) completed review forms relating to 179 unexpected deaths and 53 unexpected survivors. The healthcare system was judged to have potentially contributed to 10/179 (8%) of the unexpected deaths and 26/53 (49%) of the unexpected survivors. Failure of the model to appropriately predict risk was judged to be responsible for 135/179 (75%) of the unexpected deaths and 2/53 (4%) of the unexpected survivors. Some 10/53 (19%) of the unexpected survivors died within a few months of the 7-day period of model prediction.

Conclusions

We found little evidence that deaths occurring in patients with a low predicted mortality from risk-adjustment could be attributed to the quality of healthcare provided.
Methodological proposal for implementing an intervention to prevent pedestrian injuries, a multidisciplinary approach: the case of Cuernavaca, Morelos, Mexico
Authors: Rodríguez Jorge M , Híjar Medina Martha , Campuzano Julio César , Bangdiwala Shrikant I , Villaveces Andrés
Published: April 2013

Objective

To describe the process for implementing an intervention to prevent pedestrian injuries in Cuernavaca, Mexico, using a multidisciplinary and evidence-based approach.

Methodology

This study included two phases with several stages. The first was an overall assessment of pedestrian mortality in Cuernavaca that included a comparison of pedestrian mortality between Cuernavaca and the rest of Morelos State (1998–2007), an analysis and geocoding of pedestrian injuries (2008–2009), and a description of techniques for selecting and analysing clustered events using road audits. The second phase focused on selecting an intervention through a review of the literature seeking the ‘best evidence’ adaptable to the local context and a methodological evaluation for implementation including techniques for cluster randomisation and for the evaluation of possible effects of the intervention.

Results

Between 2008 and 2009, in Cuernavaca there were 620 pedestrian injury events: 59.4% were men, the mean age was 36.3 years, 70% occurred during daylight hours, 55% had mild severity and there were no differences by sex (p>0.05). We generated evidence, that when combined with the results from a systematic search of the literature, provided sufficient information for the implementation of a pedestrian injury prevention intervention.

Conclusions

A combination of strategies and disciplines makes it possible to comprehensively address this public health problem, allowing interventions to strengthen aspects of pedestrian safety while accounting for characteristics of the local context; the promotion of local measures, of low cost and high impact, with modifications to the road infrastructure, are more effective in addressing pedestrian vulnerabilities.
Biomarkers of oxidative stress and tissue damage released by muscle and liver after a single bout of swimming exercise
Authors: Ramos Dionizio , Martins Eduarda Gabrielle , Viana-Gomes Diego , Casimiro-Lopes Gustavo , Salerno Verônica P.
Published: April 2013

Both acute exercise and excessive training can cause oxidative stress. The resulting increase in free radicals and the inadequate response from antioxidant systems can lead to a framework of cellular damage. An association between affected tissue and the biomarkers of oxidative stress that appear in plasma has not been clearly established. The aim of this study was to evaluate the source of oxidative stress biomarkers found in the plasma of untrained rats after a single bout of swimming exercise at 2 different intensities: low intensity (SBLIE) or high intensity (SBHIE). Immediately after the exercise, aspartate transaminase (AST), alanine transaminase (ALT), γ-glutamyltransferase (GGT), and lactate dehydrogenase (LDH) were measured in plasma to characterize cell damage. Oxidative stress was assessed using protein carbonylation (PC), total antioxidant capacity (TAC), and thiobarbituric acid reactive substances (TBARS) quantified by malondialdehyde concentration. SBHIE raised levels of plasma AST (93%) and ALT (17%), and both exercise regimens produced an increase in GGT (7%) and LDH (∼55%). Plasma levels of PC and TBARS were greater in the SBHIE group; there were no changes in TAC. SBLIE caused only a modest increase in TBARS. In muscle, there were no changes in TAC, PC, or TBARS, regardless of exercise intensity, In the liver, TAC and TBARS increased significantly in both the SBLIE and SBHIE groups. This indicates that the oxidative stress biomarkers measured in the plasma immediately after a single bout of swimming exercise were generated primarily in the liver, not in muscle.
Comparison of digital outcrop and conventional data collection approaches for the characterization of naturally fractured reservoir analogues
Authors: Seers Thomas D. , Hodgetts David
Published: April 2013

In this study, fracture systems developed within faulted, high-porosity sandstones in the decommissioned mines of Alderley Edge, Cheshire, UK are characterized using lidar (Light Detection And Ranging)-based analysis. The geometry of the mine workings prove to be conducive to the extraction of fracture attributes, whilst providing a degree of exposure of a notable Triassic-aged reservoir outcrop analogue (Helsby Sandstone Formation) not afforded at the surface.

To test the fidelity of the approach, fracture statistics generated from lidar-derived digital outcrop models are compared to an equivalent dataset collected using conventional manual surveys, with digital outcrop and manually acquired fracture attributes used to populate discrete fracture network models. These are upscaled to provide equivalent porous medium properties, enabling the impact of uncertainties introduced into fracture modelling workflows by lidar-based techniques to be assessed.

Whilst broadly comparable to fracture attributes obtained using manual surveys, the systematic underrepresentation of fracture properties is observed within lidar-derived dataset, resulting in the underestimation of fracture network flow capacity. The study results suggest that, whilst enhancing data acquisition rates and coverage of exposure surfaces, the use of digital discontinuity analysis may introduce additional biases into fracture datasets, increasing the level of uncertainty within resultant modelled networks.
Landscape evolution and ice-sheet behaviour in a semi-arid polar environment: James Ross Island, NE Antarctic Peninsula
Authors: Davies Bethan J. , Glasser Neil F. , Carrivick Jonathan L. , Hambrey Michael J. , Smellie John L. , Nývlt Daniel
Published: April 2013

This study of landscape evolution presents both new modern and palaeo process-landform data, and analyses the behaviour of the Antarctic Peninsula Ice Sheet through the Last Glacial Maximum (LGM), the Holocene and to the present day. Six sediment-landform assemblages are described and interpreted for Ulu Peninsula, James Ross Island, NE Antarctic Peninsula: (1) the Glacier Ice and Snow Assemblage; (2) the Glacigenic Assemblage, which relates to LGM sediments and comprises both erratic-poor and erratic-rich drift, deposited by cold-based and wet-based ice and ice streams respectively; (3) the Boulder Train Assemblage, deposited during a Mid-Holocene glacier readvance; (4) the Ice-cored Moraine Assemblage, found in front of small cirque glaciers; (5) the Paraglacial Assemblage including scree, pebble-boulder lags, and littoral and fluvial processes; and (6) the Periglacial Assemblage including rock glaciers, protalus ramparts, blockfields, solifluction lobes and extensive patterned ground. The interplay between glacial, paraglacial and periglacial processes in this semi-arid polar environment is important in understanding polygenetic landforms. Crucially, cold-based ice was capable of sediment and landform genesis and modification. This landsystem model can aid the interpretation of past environments, but also provides new data to aid the reconstruction of the last ice sheet to overrun James Ross Island.
The Offshore New Harbour Project: deciphering the Middle Miocene through Late Eocene seismic stratigraphy of Offshore New Harbour, western Ross Sea, Antarctica
Authors: Pekar Stephen F. , Speece Marvin A. , Wilson Gary S. , Sunwall David S. , Tinto Kirsty J.
Published: April 2013

In the Austral spring of 2008, over 48 km of multi-channel seismic data were collected offshore of New Harbour, which is located in the western most Ross Sea, Antarctica. This project is part of the ANDRILL (Antarctic Geological Drilling) Programme's Offshore New Harbour Project, with its aim to investigate the stratigraphic and tectonic history of the inner shelf of southern McMurdo Sound. Correlating the seismic data to CIROS-1 and ANDRILL AND-2A drillholes provided age dates ranging from Late Miocene to at least Late Eocene for the reflectors and lithological descriptions for the seismic units. This permitted development of a glacial history of this area, resulting in dividing the seismic data into three units: an Upper Eocene–Lower Oligocene unit; an Upper Oligocene–Lower Miocene unit; and a Middle Miocene and younger unit. The seismic section below 700 mbsf has two possible interpretations: (1) substantial Eocene strata are present downdip of CIROS-1, which would represent a potential future-drilling objective for the ANDRILL Programme; or (2) these reflectors may be equivalent to Devonian strata recovered at the Cape Roberts 3 as the two-way traveltime of Eocene and Devonian strata are nearly identical, making it difficult to discriminate between them in seismic data.