bluuebunny/crossref_metadata_2025 huggingface dataset:
Resistance training increases skeletal muscle oxidative capacity and net intramuscular triglyceride breakdown in type I and II fibres of sedentary males
Authors: Shepherd S. O. , Cocks M. , Tipton K. D. , Witard O. C. , Ranasinghe A. M. , Barker T. A. , Wagenmakers A. J. M. , Shaw C. S.
Published: April 2014

New Findings

What is the central question of this study?

Recent research from our laboratory, supported by in vitro effects of perilipins, suggested that improvements in insulin sensitivity following endurance training are mechanistically linked to increases in muscle oxidative capacity, intramuscular triglyceride utilization during moderate endurance exercise and increases in the content of the lipid droplet‐associated perilipins 2 and 5. This study aimed to investigate whether these adaptations also occur in response to resistance training.

What is the main finding and its importance?

Six weeks of resistance training increased all the mentioned variables. These novel data suggest that improvements in muscle oxidative capacity and lipid metabolism contribute to the increase in insulin sensitivity following resistance training.

Recent in vitro and in vivo experimental observations suggest that improvements in insulin sensitivity following endurance training are mechanistically linked to increases in muscle oxidative capacity, intramuscular triglyceride (IMTG) utilization during endurance exercise and increases in the content of the lipid droplet‐associated perilipin 2 (PLIN2) and perilipin 5 (PLIN5). This study investigated the hypothesis that similar adaptations may also underlie the resistance training (RT)‐induced improvements in insulin sensitivity. Thirteen sedentary men (20 ± 1 years old; body mass index 24.8 ± 0.8 kg m−2) performed 6 weeks of whole‐body RT (three times per week), and changes in peak O2 uptake (in millilitres per minute per kilogram) and insulin sensitivity were assessed. Muscle biopsies (n = 8) were obtained before and after 60 min steady‐state cycling at ∼65% peak O2 uptake. Immunofluorescence microscopy was used to assess changes in oxidative capacity (measured as cytochrome c oxidase protein content), IMTG and PLIN2 and PLIN5 protein content. Resistance training increased peak O2 uptake (by 8 ± 3%), COX protein content (by 46 ± 13 and 61 ± 13% in type I and II fibres, respectively) and the Matsuda insulin sensitivity index (by 47 ± 6%; all P < 0.05). In type I fibres, IMTG (by 52 ± 11%; P < 0.05) and PLIN2 content (by 107 ± 19%; P < 0.05) were increased and PLIN5 content tended to increase (by 54 ± 22%; P = 0.054) post‐training. In type II fibres, PLIN2 content increased (by 57 ± 20%; P < 0.05) and IMTG (by 46 ± 17%; P = 0.1) and PLIN5 content (by 44 ± 24%; P = 0.054) tended to increase post‐training. Breakdown of IMTG during moderate‐intensity exercise was greater in both type I and type II fibres (by 43 ± 5 and 37 ± 5%, respectively; P < 0.05) post‐RT. The results confirm the hypothesis that RT enhances muscle oxidative capacity and increases IMTG breakdown and the content of PLIN2 and PLIN5 in both type I and type II fibres during endurance‐type exercise.
Mass and Density Measurements of Live and Dead Gram-Negative and Gram-Positive Bacterial Populations
Authors: Lewis Christina L. , Craig Caelli C. , Senecal Andre G.
Published: April 2014

Monitoring cell growth and measuring physical features of food-borne pathogenic bacteria are important for better understanding the conditions under which these organisms survive and proliferate. To address this challenge, buoyant masses of live and dead
Escherichia coli
O157:H7 and
Listeria innocua
were measured using Archimedes, a commercially available suspended microchannel resonator (SMR). Cell growth was monitored with Archimedes by observing increased cell concentration and buoyant mass values of live growing bacteria. These growth data were compared to optical density measurements obtained with a Bioscreen system. We observed buoyant mass measurements with Archimedes at cell concentrations between 10
5
and 10
8
cells/ml, while growth was not observed with optical density measurements until the concentration was 10
7
cells/ml. Buoyant mass measurements of live and dead cells with and without exposure to hydrogen peroxide stress were also compared; live cells generally had a larger buoyant mass than dead cells. Additionally, buoyant mass measurements were used to determine cell density and total mass for both live and dead cells. Dead
E. coli
cells were found to have a larger density and smaller total mass than live
E. coli
cells. In contrast, density was the same for both live and dead
L. innocua
cells, while the total mass was greater for live than for dead cells. These results contribute to the ongoing challenge to further develop existing technologies used to observe cell populations at low concentrations and to measure unique physical features of cells that may be useful for developing future diagnostics.
Transcription of Two Adjacent Carbohydrate Utilization Gene Clusters in Bifidobacterium breve UCC2003 Is Controlled by LacI- and Repressor Open Reading Frame Kinase (ROK)-Type Regulators
Authors: O'Connell Kerry Joan , O'Connell Motherway Mary , Liedtke Andrea , Fitzgerald Gerald F. , Ross R. Paul , Stanton Catherine , Zomer Aldert , van Sinderen Douwe
Published: April 2014

Members of the genus
Bifidobacterium
are commonly found in the gastrointestinal tracts of mammals, including humans, where their growth is presumed to be dependent on various diet- and/or host-derived carbohydrates. To understand transcriptional control of bifidobacterial carbohydrate metabolism, we investigated two genetic carbohydrate utilization clusters dedicated to the metabolism of raffinose-type sugars and melezitose. Transcriptomic and gene inactivation approaches revealed that the raffinose utilization system is positively regulated by an activator protein, designated RafR. The gene cluster associated with melezitose metabolism was shown to be subject to direct negative control by a LacI-type transcriptional regulator, designated MelR1, in addition to apparent indirect negative control by means of a second LacI-type regulator, MelR2.
In silico
analysis, DNA-protein interaction, and primer extension studies revealed the MelR1 and MelR2 operator sequences, each of which is positioned just upstream of or overlapping the correspondingly regulated promoter sequences. Similar analyses identified the RafR binding operator sequence located upstream of the
rafB
promoter. This study indicates that transcriptional control of gene clusters involved in carbohydrate metabolism in bifidobacteria is subject to conserved regulatory systems, representing either positive or negative control.
Population Structure of Listeria monocytogenes Serotype 4b Isolates from Sporadic Human Listeriosis Cases in the United States from 2003 to 2008
Authors: Lee Sangmi , Ward Todd J. , Graves Lewis M. , Tarr Cheryl L. , Siletzky Robin M. , Kathariou Sophia
Published: April 2014

Listeria monocytogenes
can cause severe food-borne disease (listeriosis). Numerous outbreaks have involved three serotype 4b epidemic clones (ECs): ECI, ECII, and ECIa. However, little is known about the population structure of
L. monocytogenes
serotype 4b from sporadic listeriosis in the United States, even though most cases of human listeriosis are in fact sporadic. Here we analyzed 136 serotype 4b isolates from sporadic cases in the United States, 2003 to 2008, utilizing multiple tools including multilocus genotyping, pulsed-field gel electrophoresis, and sequence analysis of the
inlAB
locus. ECI, ECII, and ECIa were frequently encountered (32, 17, and 7%, respectively). However, annually 30 to 68% of isolates were outside these ECs, and several novel clonal groups were identified. An estimated 33 and 17% of the isolates, mostly among the ECs, were resistant to cadmium and arsenic, respectively, but resistance to benzalkonium chloride was uncommon (3%) among the sporadic isolates. The frequency of clonal groups fluctuated within the 6-year study period, without consistent trends. However, on several occasions, temporal clusters of isolates with indistinguishable genotypes were detected, suggesting the possibility of hidden multistate outbreaks. Our analysis suggests a complex population structure of serotype 4b
L. monocytogenes
from sporadic disease, with important contributions by ECs and several novel clonal groups. Continuous monitoring will be needed to assess long-term trends in clonality patterns and population structure of
L. monocytogenes
from sporadic listeriosis.
Insects Represent a Link between Food Animal Farms and the Urban Environment for Antibiotic Resistance Traits
Authors: Zurek Ludek , Ghosh Anuradha
Published: April 2014

Antibiotic-resistant bacterial infections result in higher patient mortality rates, prolonged hospitalizations, and increased health care costs. Extensive use of antibiotics as growth promoters in the animal industry represents great pressure for evolution and selection of antibiotic-resistant bacteria on farms. Despite growing evidence showing that antibiotic use and bacterial resistance in food animals correlate with resistance in human pathogens, the proof for direct transmission of antibiotic resistance is difficult to provide. In this review, we make a case that insects commonly associated with food animals likely represent a direct and important link between animal farms and urban communities for antibiotic resistance traits. Houseflies and cockroaches have been shown to carry multidrug-resistant clonal lineages of bacteria identical to those found in animal manure. Furthermore, several studies have demonstrated proliferation of bacteria and horizontal transfer of resistance genes in the insect digestive tract as well as transmission of resistant bacteria by insects to new substrates. We propose that insect management should be an integral part of pre- and postharvest food safety strategies to minimize spread of zoonotic pathogens and antibiotic resistance traits from animal farms. Furthermore, the insect link between the agricultural and urban environment presents an additional argument for adopting prudent use of antibiotics in the food animal industry.
A Mesophilic, Autotrophic, Ammonia-Oxidizing Archaeon of Thaumarchaeal Group I.1a Cultivated from a Deep Oligotrophic Soil Horizon
Authors: Jung Man-Young , Park Soo-Je , Kim So-Jeong , Kim Jong-Geol , Sinninghe Damsté Jaap S. , Jeon Che Ok , Rhee Sung-Keun
Published: April 2014

Soil nitrification plays an important role in the reduction of soil fertility and in nitrate enrichment of groundwater. Various ammonia-oxidizing archaea (AOA) are considered to be members of the pool of ammonia-oxidizing microorganisms in soil. This study reports the discovery of a chemolithoautotrophic ammonia oxidizer that belongs to a distinct clade of nonmarine thaumarchaeal group I.1a, which is widespread in terrestrial environments. The archaeal strain MY2 was cultivated from a deep oligotrophic soil horizon. The similarity of the 16S rRNA gene sequence of strain MY2 to those of other cultivated group I.1a thaumarchaeota members, i.e.,
Nitrosopumilus maritimus
and “
Candidatus
Nitrosoarchaeum koreensis,” is 92.9% for both species. Extensive growth assays showed that strain MY2 is chemolithoautotrophic, mesophilic (optimum temperature, 30°C), and neutrophilic (optimum pH, 7 to 7.5). The accumulation of nitrite above 1 mM inhibited ammonia oxidation, while ammonia oxidation itself was not inhibited in the presence of up to 5 mM ammonia. The genome size of strain MY2 was 1.76 Mb, similar to those of
N. maritimus
and “
Ca
. Nitrosoarchaeum koreensis,” and the repertoire of genes required for ammonia oxidation and carbon fixation in thaumarchaeal group I.1a was conserved. A high level of representation of conserved orthologous genes for signal transduction and motility in the noncore genome might be implicated in niche adaptation by strain MY2. On the basis of phenotypic, phylogenetic, and genomic characteristics, we propose the name “
Candidatus
Nitrosotenuis chungbukensis” for the ammonia-oxidizing archaeal strain MY2.
Environmental Transmission of Human Noroviruses in Shellfish Waters
Authors: Campos Carlos J. A. , Lees David N.
Published: April 2014

Human noroviruses (NoV) are the most common cause of epidemic gastroenteritis following consumption of bivalve shellfish contaminated with fecal matter. NoV levels can be effectively reduced by some sewage treatment processes such as activated sludge and membrane bioreactors. However, tertiary sewage treatment and substantial sewage dilution are usually required to achieve low concentrations of virus in shellfish. Most outbreaks have been associated with shellfish harvested from waters affected by untreated sewage from, for example, storm overflows or overboard disposal of feces from boats. In coastal waters, NoV can remain in suspension or associate with organic and inorganic matter and be accumulated by shellfish. Shellfish take considerably longer to purge NoV than fecal indicator bacteria when transferred from sewage-polluted estuarine waters to uncontaminated waters. The abundance and distribution of NoV in shellfish waters are influenced by the levels of sewage treatment, proximity of shellfish beds to sewage sources, rainfall, river flows, salinity, and water temperature. Detailed site-specific information on these factors is required to design measures to control the viral risk.
Role of
norEF
in Denitrification, Elucidated by Physiological Experiments with Rhodobacter sphaeroides
Authors: Bergaust Linda L. , Hartsock Angela , Liu Binbin , Bakken Lars R. , Shapleigh James P.
Published: April 2014

Many denitrifying organisms contain the
norEF
gene cluster, which codes for two proteins that are thought to be involved in denitrification because they are expressed during the reduction of nitrite and nitric oxide. The products of both genes are predicted to be membrane associated, and the
norE
product is a member of the cytochrome
c
oxidase subunit III family. However, the specific role of
norEF
is unknown. The denitrification phenotypes of
Rhodobacter sphaeroides
strains with and without
norEF
genes were studied, and it was found that loss of
norEF
lowered the rate of denitrification from nitrate and resulted in accumulation of micromolar concentrations of nitric oxide during denitrification from nitrite.
norEF
appears to have no direct role in the reduction of nitric oxide; however, since deletion of
norEF
in the wild-type 2.4.3 strain had essentially no influence on the kinetics of potential nitric oxide reduction (
V
max
and

K
s

), as measured by monitoring the depletion of a bolus of nitric oxide injected into anoxic cultures without any other electron acceptors. However,
norEF
-deficient cells that had undergone a more chronic exposure to micromolar concentrations of nitric oxide showed an ∼50% reduction in
V
max
but no change in apparent

K
s

. These results can explain the occurrence of
norEF
in the 2.4.3 strain of
R. sphaeroides
, which can reduce nitrate to nitrous oxide, and their absence from strains such as 2.4.1, which likely use nitric oxide reductase to mitigate stress due to episodic exposure to nitric oxide from exogenous sources.
Influence of Iron and Aeration on Staphylococcus aureus Growth, Metabolism, and Transcription
Authors: Ledala Nagender , Zhang Bo , Seravalli Javier , Powers Robert , Somerville Greg A.
Published: April 2014

Staphylococcus aureus
is a prominent nosocomial pathogen and a major cause of biomaterial-associated infections. The success of
S. aureus
as a pathogen is due in part to its ability to adapt to stressful environments. As an example, the transition from residing in the nares to residing in the blood or deeper tissues is accompanied by changes in the availability of nutrients and elements such as oxygen and iron. As such, nutrients, oxygen, and iron are important determinants of virulence factor synthesis in
S. aureus
. In addition to influencing virulence factor synthesis, oxygen and iron are critical cofactors in enzymatic and electron transfer reactions; thus, a change in iron or oxygen availability alters the bacterial metabolome. Changes in metabolism create intracellular signals that alter the activity of metabolite-responsive regulators such as CodY, RpiRc, and CcpA. To assess the extent of metabolomic changes associated with oxygen and iron limitation,
S. aureus
cells were cultivated in iron-limited medium and/or with decreasing aeration, and the metabolomes were examined by nuclear magnetic resonance (NMR) spectroscopy. As expected, oxygen and iron limitation dramatically decreased tricarboxylic acid (TCA) cycle activity, creating a metabolic block and significantly altering the metabolome. These changes were most prominent during post-exponential-phase growth, when TCA cycle activity was maximal. Importantly, many of the effects of iron limitation were obscured by aeration limitation. Aeration limitation not only obscured the metabolic effects of iron limitation but also overrode the transcription of iron-regulated genes. Finally, in contrast to previous speculation, we confirmed that acidification of the culture medium occurs independent of the availability of iron.
Genetic Dissection of T4 Lysis
Authors: Moussa Samir H. , Lawler Jessica L. , Young Ry
Published: April 2014

t
is the holin gene for coliphage T4, encoding a 218-amino-acid (aa) protein essential for the inner membrane hole formation that initiates lysis and terminates the phage infection cycle. T is predicted to be an integral membrane protein that adopts an N
in
-C
out
topology with a single transmembrane domain (TMD). This holin topology is different from those of the well-studied holins S105 (3 TMDs; N
out
-C
in
) of the coliphage lambda and S68 (2 TMDs; N
in
-C
in
) of the lambdoid phage 21. Here, we used random mutagenesis to construct a library of lysis-defective alleles of
t
to discern residues and domains important for holin function and for the inhibition of lysis by the T4 antiholin, RI. The results show that mutations in all 3 topological domains (N-terminal cytoplasmic, TMD, and C-terminal periplasmic) can abrogate holin function. Additionally, several lysis-defective alleles in the C-terminal domain are no longer competent in binding RI. Taken together, these results shed light on the roles of the previously uncharacterized N-terminal and C-terminal domains in lysis and its real-time regulation.
EscO, a Functional and Structural Analog of the Flagellar FliJ Protein, Is a Positive Regulator of EscN ATPase Activity of the Enteropathogenic Escherichia coli Injectisome
Authors: Romo-Castillo Mariana , Andrade Angel , Espinosa Norma , Monjarás Feria Julia , Soto Eduardo , Díaz-Guerrero Miguel , González-Pedrajo Bertha
Published: April 2014

Type III secretion systems (T3SSs) are multiprotein molecular devices used by many Gram-negative bacterial pathogens to translocate effector proteins into eukaryotic cells. A T3SS is also used for protein export in flagellar assembly, which promotes bacterial motility. The two systems are evolutionarily related, possessing highly conserved components in their export apparatuses. Enteropathogenic
Escherichia coli
(EPEC) employs a T3SS, encoded by genes in the locus of enterocyte effacement (LEE) pathogenicity island, to colonize the human intestine and cause diarrheal disease. In the present work, we investigated the role of the LEE-encoded EscO protein (previously Orf15 or EscA) in T3SS biogenesis. We show that EscO shares similar properties with the flagellar FliJ and the
Yersinia
YscO protein families. Our findings demonstrate that EscO is essential for secretion of all categories of T3SS substrates. Consistent with its central role in protein secretion, it was found to interact with the ATPase EscN and its negative regulator, EscL, of the export apparatus. Moreover, we show that EscO stimulates EscN enzymatic activity; however, it is unable to upregulate ATP hydrolysis in the presence of EscL. Remarkably, EscO partially restored the swimming defect of a
Salmonella
flagellar
fliJ
mutant and was able to stimulate the ATPase activity of FliI. Overall, our data indicate that EscO is the virulence counterpart of the flagellar FliJ protein.
Analysis of Factors That Affect FlgM-Dependent Type III Secretion for Protein Purification with Salmonella enterica Serovar Typhimurium
Authors: Guo Shukui , Alshamy Israa , Hughes Kelly T. , Chevance Fabienne F. V.
Published: April 2014

The FlgM protein is secreted in response to flagellar hook-basal body secretion and can be used as a secretion signal to direct selected protein secretion via the flagellar type III secretion (T3S) system [H. M. Singer, M. Erhardt, A. M. Steiner, M. M. Zhang, D. Yoshikami, G. Bulaj, B. M. Olivera, and K. T. Hughes, mBio 3(3):e00115-12, 2012,
http://dx.doi.org/10.1128/mBio.00115-12
]. Conditions known to affect flagellar gene expression, FlgM stability, and flagellar T3S were tested either alone or in combination to determine their effects on levels of secreted FlgM. These conditions included mutations that affect activity of the flagellar FlhD
4
C
2
master regulatory protein complex or the FlgM T3S chaperone σ
28
, the removal of
Salmonella p
athogenicity
i
sland 1 (Spi1), the removal of flagellar late secretion substrates that could compete with FlgM for secretion, and changes in the ionic strength of the growth medium. Conditions that enhanced FlgM secretion were combined in order to maximize levels of secreted FlgM. An optimized FlgM secretion strain was used to secrete and isolate otherwise difficult-to-produce proteins and peptides fused to the C terminus of FlgM. These include cysteine-rich, hydrophobic peptides (conotoxins δ-SVIE and MrVIA), nodule-specific, cysteine-rich antimicrobial peptides (NCR), and a malaria surface antigen domain of apical membrane antigen AMA-1.
FlgN Is Required for Flagellum-Based Motility by Bacillus subtilis
Authors: Cairns Lynne S. , Marlow Victoria L. , Kiley Taryn B. , Birchall Christopher , Ostrowski Adam , Aldridge Phillip D. , Stanley-Wall Nicola R.
Published: April 2014

The assembly of the bacterial flagellum is exquisitely controlled. Flagellar biosynthesis is underpinned by a specialized type III secretion system that allows export of proteins from the cytoplasm to the nascent structure.
Bacillus subtilis
regulates flagellar assembly using both conserved and species-specific mechanisms. Here, we show that YvyG is essential for flagellar filament assembly. We define YvyG as an orthologue of the
Salmonella enterica
serovar Typhimurium type III secretion system chaperone, FlgN, which is required for the export of the hook-filament junction proteins, FlgK and FlgL. Deletion of
flgN
(
yvyG
) results in a nonmotile phenotype that is attributable to a decrease in
hag
translation and a complete lack of filament polymerization. Analyses indicate that a
flgK-flgL
double mutant strain phenocopies deletion of
flgN
and that overexpression of
flgK-flgL
cannot complement the motility defect of a Δ
flgN
strain. Furthermore, in contrast to previous work suggesting that phosphorylation of FlgN alters its subcellular localization, we show that mutation of the identified tyrosine and arginine FlgN phosphorylation sites has no effect on motility. These data emphasize that flagellar biosynthesis is differentially regulated in
B. subtilis
from classically studied Gram-negative flagellar systems and questions the biological relevance of some posttranslational modifications identified by global proteomic approaches.
The LacI-Type Transcriptional Regulator AraR Acts as an
l
-Arabinose-Responsive Repressor of
l
-Arabinose Utilization Genes in Corynebacterium glutamicum ATCC 31831
Authors: Kuge Takayuki , Teramoto Haruhiko , Yukawa Hideaki , Inui Masayuki
Published: April 2014

The
Corynebacterium glutamicum
ATCC 31831
araBDA
operon consists of three
l
-arabinose catabolic genes, upstream of which the
galM
,
araR
, and
araE
genes are located in opposite orientation.
araR
encodes a LacI-type transcriptional regulator that negatively regulates the
l
-arabinose-inducible expression of
araBDA
and
araE
(encoding an
l
-arabinose transporter), through a mechanism that has yet to be identified. Here we show that the AraR protein binds
in vitro
to three sites: one upstream of
araBDA
and two upstream of
araE
. We verify that a 16-bp consensus palindromic sequence is essential for binding of AraR, using a series of mutations introduced upstream of
araB
in electrophoretic mobility shift assays. Moreover, the DNA-binding activity of AraR is reduced by
l
-arabinose. We employ quantitative reverse transcription-PCR (qRT-PCR) analyses using various mutant strains deficient in
l
-arabinose utilization genes to demonstrate that the prominent upregulation of
araBDA
and
araE
within 5 min of
l
-arabinose supplementation is dependent on the uptake but independent of the catabolism of
l
-arabinose. Similar expression patterns, together with the upregulation by
araR
disruption without
l
-arabinose, are evident with the apparent
galM-araR
operon, although attendant changes in expression levels are much smaller than those realized with the expression of
araBDA
and
araE
. The AraR-binding site upstream of
araB
overlaps the −10 region of the divergent
galM
promoter. These observations indicate that AraR acts as a transcriptional repressor of
araBDA
,
araE
, and
galM-araR
and that
l
-arabinose acts as an intracellular negative effector of the AraR-dependent regulation.
A Proposed Genus Boundary for the Prokaryotes Based on Genomic Insights
Authors: Qin Qi-Long , Xie Bin-Bin , Zhang Xi-Ying , Chen Xiu-Lan , Zhou Bai-Cheng , Zhou Jizhong , Oren Aharon , Zhang Yu-Zhong
Published: April 2014

Genomic information has already been applied to prokaryotic species definition and classification. However, the contribution of the genome sequence to prokaryotic genus delimitation has been less studied. To gain insights into genus definition for the prokaryotes, we attempted to reveal the genus-level genomic differences in the current prokaryotic classification system and to delineate the boundary of a genus on the basis of genomic information. The average nucleotide sequence identity between two genomes can be used for prokaryotic species delineation, but it is not suitable for genus demarcation. We used the percentage of conserved proteins (POCP) between two strains to estimate their evolutionary and phenotypic distance. A comprehensive genomic survey indicated that the POCP can serve as a robust genomic index for establishing the genus boundary for prokaryotic groups. Basically, two species belonging to the same genus would share at least half of their proteins. In a specific lineage, the genus and family/order ranks showed slight or no overlap in terms of POCP values. A prokaryotic genus can be defined as a group of species with all pairwise POCP values higher than 50%. Integration of whole-genome data into the current taxonomy system can provide comprehensive information for prokaryotic genus definition and delimitation.
Improving the accessibility of trust guidelines for junior doctors at North Bristol NHS Trust
Authors: Tarrant Madelaine , Honeyman Calum , Aquilina Alex , Young Katie
Published: April 2014

Medicine is becoming increasingly protocol driven. This provides a standardised format for doctors to deliver best practice, especially in the acute setting.

40 junior doctors were asked to locate three Trust guidelines: venous thromboembolism (VTE) prophylaxis; antibiotic prescribing; and management of upper gastrointestinal bleed (UGIB). For each doctor the time taken and number of mouse clicks to access each guideline was recorded. Following successful redesign of the Trust intranet we completed a re-audit.

Initial results showed 48% of doctors were unable to locate the UGIB or the VTE guidelines within 5 min. For those who were able to locate the guidelines it took an average of 111 sec and 17 mouse clicks. 100% of doctors were able to locate the antibiotic guidelines in 12 sec and with two clicks. These are accessible via a single port of access.

Following our redesign of the Trust intranet 100% of doctors located all three guidelines in an average time of 7.2 sec and in 2.1 clicks. Improvement in access to VTE prophylaxis and UGIB was statistically significant (p=0.001).

Redesigning our Trust intranet homepage has significantly improved the accessibility of acute surgical and medical guidelines.
Obesity management in a Paediatric Emergency Department
Authors: Knight Martin , Booth Charlotte
Published: April 2014

Increasing numbers of children are either overweight or obese, which has a significant negative impact on their current quality of life and their future health. The results of England's National Child Measurement Programme 2012-13 revealed that approximately one fifth of 4- to 5-year-olds and one third of children aged 10 to 11 years were overweight or obese. Emergency departments (EDs) see large numbers of children every year and therefore represent a potential opportunity to address the obesity issue.

A pathway was devised to guide referral of overweight or obese children attending the ED to an existing local healthy living programme. Training sessions on the referral process were provided to ED staff. Audits of ED attendance records were undertaken before and after the implementation of the pathway, and the local programme uptake rate for the first 25 referrals was assessed.

Of the first 25 children referred, two (8%) had participated in a programme, six (24%) were on a waiting list, and four (16%) were referred on to services in their local area, giving an overall engagement rate of 48%.

Further work needs to be undertaken to increase referrals; however, addressing childhood obesity by accessing existing local services from the ED remains a valuable endeavour.
Prehospital use of furosemide for the treatment of heart failure
Authors: Pan Andy , Stiell Ian G , Dionne Richard , Maloney Justin
Published: April 2014

Introduction

The diagnosis and management of acute decompensated heart failure (HF) in the prehospital setting can be challenging. The objectives of this study are to evaluate the appropriateness of furosemide use by Emergency Medical Services (EMS) and its association with adverse outcomes.

Methods

This study was a multi-centre health records review of EMS patients who received prehospital furosemide or had an emergency department (ED) diagnosis of HF. We included acutely ill patients ≥50 years of age with shortness of breath transported by land EMS. Univariate and logistic regression analyses were performed to determine associations between furosemide use and serious adverse outcomes (acute renal failure, intubation, vasopressors or death).

Results

The study population consisted of 330 patients (N=58, furosemide given by EMS but no HF diagnosed in ED; N=110, furosemide given, HF diagnosed; N=162, no furosemide given, HF diagnosed). The median dose of intravenous furosemide was 80 mg (range 20–80 mg). Serious adverse outcomes occurred in 61 patients (19.0%, 23.6% and 14.8% of the three groups, respectively; p=0.18). The adjusted ORs for adverse events with furosemide use was 0.62 (95% CI 0.33 to 1.43) in patients with a diagnosis of HF and 1.14 (95% CI 0.58 to 2.23) in those without.

Conclusions

More than a third of patients who received prehospital furosemide did not have an HF diagnosis, suggesting that the prehospital diagnosis of HF can be challenging. Serious adverse outcomes were identified in all patient groups and we found no statistically significant associations between furosemide use and adverse events.
Enquiry time as part of turnaround time: when do our clinicians really consult our results?
Authors: Rodríguez-Borja Enrique , Villalba-Martínez C , Carratalá-Calvo A
Published: April 2014

Background

Traditionally, laboratories’ turnaround times (TAT) have been calculated by only considering analytical or intralaboratory steps. The measure of the postanalytical impact in TAT has barely been studied and, more specifically, the running time from when finalised results are available to when clinicians make their first enquiry with an electronic medical record.

Methods

During May–June of 2013, two ‘Times’ were collected from our laboratory information system for all the priority requests coming from our day hospitals: ‘Validation time’ (TV), as the request report time with full verified results and ‘Enquiry time’ (TQ), as the time when the first consult was made via electronic medical record. We classified requests in groups depending on time results, and TQ-TV (percentiles) were calculated for each group.

Results

654 (69%) requests were consulted by clinicians before 15 : 00 on the same day with available results. 191 (20%) were consulted after 15 : 00 and had complete results as well (p50 (TQ–TV): 5 days) while 61 (7%) were never consulted (up to 31/12/2013). 39 (4%) requests were finally consulted before 15 : 00 h with no available results, but the average time difference between validation and enquiry was 31 min.

Conclusions

The results obtained lead us to reconsider the TAT established with our day hospitals in order to know if priority has to be reviewed or if there are failures in follow-up results. ‘Enquiry time’ appears to be a powerful tool in detecting these issues and shows that TATs are no longer just a ‘laboratory problem’.
Survey of tail injuries sustained by working gundogs and terriers in Scotland
Authors: Lederer R. , Bennett D. , Parkin T.
Published: April 2014

Working dog owners in Scotland were invited to take part in an internet survey regarding the 2010/2011 shooting season, which was designed to estimate the prevalence of tail injuries; assess the risk of tail injuries in docked and undocked working dogs; and identify risk factors for owner‐reported tail injuries. Of 2860 working dogs, 13.5 per cent sustained at least one tail injury during the 2010/2011 shooting season. Undocked spaniels and hunt point retrievers (HPRs) were at greatest risk of tail injury with 56.6 per cent of undocked spaniels and 38.5 per cent of undocked HPRs sustaining at least one tail injury during the season. There was no statistically significant difference in the risk of tail injury in dogs with tails docked by one‐third, half or shorter. To prevent one tail injury in one shooting season, between two and 18 spaniels or HPRs would need to be docked as puppies. The authors believe that this work provides the best available evidence on which to base a consultation for changes to the legislation on tail docking in working dogs in Scotland. Docking the tails of HPRs and spaniels by one‐third would significantly decrease the risk of tail injury sustained while working in these breeds.
The prevalence of tail injuries in working and non‐working breed dogs visiting veterinary practices in Scotland
Authors: Cameron N. , Lederer R. , Bennett D. , Parkin T.
Published: April 2014

The aim of this paper was to estimate the prevalence of tail injuries that required veterinary examination in different breeds of dog in Scotland. The study population included all dogs that had visited one of 16 veterinary practices located in Scotland between 2002 and early 2012. The overall prevalence of tail injuries in dogs visiting one of the 16 veterinary practices was 0.59 per cent. The prevalence of tail injuries in dogs of working breeds was estimated to be 0.90 per cent. Working dog breeds that were examined by a veterinary surgeon were at a significantly greater risk of sustaining a tail injury than non‐working breeds (P<0.001). To prevent one such tail injury in these working breeds approximately 232 dogs would need to be docked as puppies. To prevent one tail amputation in spaniels, 320 spaniel puppies would need to be docked. Spaniels presented after January 2009 were 2.3 times more likely to have a tail injury than those presented before April 29, 2007 (date of the legislation that banned tail docking in Scotland). Given the results of this and the accompanying paper it may be appropriate to consider changes to the current legislation for specific breeds of working dogs.
Evidence for exsolution of Au-ore fluids from granites crystallized in the mid-crust, Archaean Louis Lake Batholith, Wyoming
Authors: Vaughn E. S. , Ridley J. R.
Published: April 2014

The Louis Lake Batholith is contemporaneous with and sits adjacent to Archaean lode gold deposits and minor copper stockwork vein systems of the South Pass Greenstone Belt, Wyoming Province, and may thus have contributed fluid into these hydrothermal systems. The batholith is granodiorite to granite, lacks extreme fractionation, and contains evidence for early formation and later replacement of an immiscible early sulphide phase. Mafic enclaves contain anomalous concentrations of gold and copper, suggesting that metals were introduced from in-mixed mafic magma. Apatite SO
3
and Cl values imply relatively early exsolution of sulphur- and chlorine-bearing fluids during fractionation. Exsolution of possibly immiscible high- and low-salinity fluids is implied from fluid inclusion petrography, even though crystallization pressures were greater than 3 kbar. Low salinity inclusions share affinities with those of orogenic gold systems, but contain notably higher Pb and Zn concentrations. High salinity inclusions have similar elemental compositions to brines of intrusion-centred ore systems.

It is proposed that sulphide melt separated early during magma fractionation, scavenging Au and Cu, but became later destabilized, possibly because of increased
f
O
2
, releasing scavenged metals. Metals were transported from the pluton in sulphur- and chlorine-bearing hydrothermal fluids of low and high salinity, with Au being preferentially transported in the low-salinity fluid.

Supplementary material:

Whole Rock Geochemical data are available at
http://www.geolsoc.org.uk/SUP18751
.
More gap than record? Qualitative and quantitative assessment of stratigraphic gaps in a field based study, with examples from the Lower Silurian Pentamerus Beds of Shropshire, England and the Lower Ordovician Ribband Gp of County Wexford, Ireland
Authors: Crossley Joseph D. , Clark Hazel E.
Published: April 2014

To assess the extent to which it is possible to identify, characterize and quantify gaps in the stratigraphic record, two contrasting sections, the Silurian Pentamerus Beds of Shropshire, England, and the Ordovician Ribband Gp of County Wexford, Ireland, have been examined in the field. In the Pentamerus Beds the life assemblages contained in silty mudstones suggest slow to little/no deposition during fair weather periods. Interbedded lenticular calcirudites characteristically feature death assemblages of thick-shelled brachiopods and are interpreted as storm deposits, implying erosional breaks in accumulation. In contrast, the Ribband Gp's regular ribbon banding of fine sandstones, siltstones and mudstones/silty mudstones suggests a more distal shelf environment. Fodinichnial trace fossils are witness to periods of more or less continuous sedimentation. The succession may have formed during seasonal alternations of stormy (thin cross-laminated sandstone beds) and fair weather periods (laminated mudstones); though longer-term climatic variations cannot be ruled out. There is minimal evidence for hiatuses in the accumulation.
NOSIX
Authors: Yu Minlan , Wundsam Andreas , Raju Muruganantham
Published: April 2014

In spite of the standardization of the OpenFlow API, it is very difficult to write an SDN controller application that is portable (i.e., guarantees correct packet processing over a wide range of switches) and achieves good performance (i.e., fully leverages switch capabilities). This is because the switch landscape is fundamentally diverse in performance, feature set and supported APIs. We propose to address this challenge via a lightweight portability layer that acts as a rendezvous point between the requirements of controller application and the vendor knowledge of switch implementations. Above, applications specify rules in virtual flow tables annotated with semantic intents and expectations. Below, vendor specific drivers map them to optimized switch-specific rule sets. NOSIX represents a first step towards achieving both portability and good performance across a diverse set of switches.
Geometry is a major determinant of flow reversal in proximal aorta
Authors: Bensalah Mourad Z. , Bollache Emilie , Kachenoura Nadjia , Giron Alain , De Cesare Alain , Macron Laurent , Lefort Muriel , Redheuill Alban , Mousseaux Elie
Published: April 2014

The aim of this study is to quantify aortic backward flow (BF) using phase-contrast cardiovascular magnetic resonance (PC-CMR) and to study its associations with age, indexes of arterial stiffness, and geometry. Although PC-CMR blood flow studies showed a simultaneous presence of BF and forward flow (FF) in the ascending aorta (AA), the relationship between aortic flows and aging as well as arterial stiffness and geometry in healthy volunteers has never been reported. We studied 96 healthy subjects [47 women, 39 ± 15 yr old (19–79 yr)]. Aortic stiffness [arch pulse wave velocity (PWVAO), AA distensibility], geometry (AA diameter and arch length), and parameters related to AA BF and FF (volumes, peaks, and onset times) were estimated from CMR. Applanation tonometry carotid-femoral pulse-wave velocity (PWVCF), carotid augmentation index, and time to return of the reflected pressure wave were assessed. Whereas FF parameters remained unchanged, BF onset time shortened significantly ( R2 = 0.18, P < 0.0001) and BF volume and BF-to-FF peaks ratio increased significantly ( R2 = 0.38 and R2 = 0.44, respectively, P < 0.0001) with aging. These two latter BF indexes were also related to stiffness indexes (PWVCF, R2 > 0.30; PWVAO, R2 > 0.24; and distensibility, R2 > 0.20, P < 0.001), augmentation index ( R2 > 0.20, P < 0.001), and aortic geometry (AA diameter, R2 > 0.58; and arch length, R2 > 0.31, P < 0.001). In multivariate analysis, aortic diameter was the strongest independent correlate of BF beyond age effect. In conclusion, AA BF estimated using PC-CMR increased significantly in terms of magnitude and volume and appeared earlier with aging and was mostly determined by aortic geometry. Thus BF indexes could be relevant markers of subclinical arterial wall alterations.
Activation of α1B-adrenoceptors contributes to intermittent hypobaric hypoxia-improved postischemic myocardial performance via inhibiting MMP-2 activation
Authors: Gao Ling , Chen Le , Lu Zhi-Zhen , Gao Hong , Wu Lan , Chen Yi-Xiong , Zhang Cai-Mei , Jiang Yu-Kun , Jing Qing , Zhang You-Yi , Yang Huang-Tian
Published: April 2014

Inhibition of matrix metalloproteinases-2 (MMP-2) activation renders cardioprotection from ischemia/reperfusion (I/R) injury; however, the signaling pathways involved have not been fully understood. Intermittent hypobaric hypoxia (IHH) has been shown to enhance myocardial tolerance to I/R injury via triggering intrinsic adaptive responses. Here we investigated whether IHH protects the heart against I/R injury via the regulation of MMP-2 and how the MMP-2 is regulated. IHH (Po2 = 84 mmHg, 4-h/day, 4 wk) improved postischemic myocardial contractile performance, lactate dehydrogenase (LDH) release, and infarct size in isolated perfused rat hearts. Moreover, IHH reversed I/R-induced MMP-2 activation and release, disorders in the levels of MMP-2 regulators, peroxynitrite (ONOO−) and tissue inhibitor of metalloproteinase-4 (TIMP-4), and loss of the MMP-2 targets α-actinin and troponin I. This protection was mimicked, but not augmented, by a MMP inhibitor doxycycline and lost by the α1-adrenoceptor (AR) antagonist prazosin. Furthermore, IHH increased myocardial α1A-AR and α1B-AR density but not α1D-AR after I/R. Concomitantly, IHH further enhanced the translocation of PKC epsilon (PKCε) and decreased the release of mitochondrial cytochrome c due to I/R via the activation of α1B-AR but not α1A-AR or α1D-AR. IHH-conferred cardioprotection in the postischemic contractile function, LDH release, MMP-2 activation, and nitrotyrosine as well as TIMP-4 contents were mimicked but not additive by α1-AR stimulation with phenylephrine and were abolished by an α1B-AR antagonist chloroethylclonidine and a PKCε inhibitor PKCε V1–2. These findings demonstrate that IHH exerts cardioprotection through attenuating excess ONOO− biosynthesis and TIMP-4 loss and sequential MMP-2 activation via the activation of α1B-AR/PKCε pathway.
Mutations flanking the carbohydrate binding site of surfactant protein D confer antiviral activity for pandemic influenza A viruses
Authors: Nikolaidis Nikolaos M. , White Mitchell R. , Allen Kimberly , Tripathi Shweta , Qi Li , McDonald Barbara , Taubenberger Jeffery , Seaton Barbara A. , McCormack Francis X. , Crouch Erika C. , Hartshorn Kevan L.
Published: April 2014

We recently reported that a trimeric neck and carbohydrate recognition domain (NCRD) fragment of human surfactant protein D (SP-D), a host defense lectin, with combinatorial substitutions at the 325 and 343 positions (D325A+R343V) exhibits markedly increased antiviral activity for seasonal strains of influenza A virus (IAV). The NCRD binds to glycan-rich viral envelope proteins including hemagglutinin (HA). We now show that replacement of D325 with serine to create D325S+R343V provided equal or increased neutralizing activity compared with D325A+R343V. The activity of the double mutants was significantly greater than that of either single mutant (D325A/S or R343V). D325A+R343V and D325S+R343V also strongly inhibited HA activity, and markedly aggregated, the 1968 pandemic H3N2 strain, Aichi68. D325S+R343V significantly reduced viral loads and mortality of mice infected with Aichi68, whereas wild-type SP-D NCRD did not. The pandemic H1N1 strains of 1918 and 2009 have only one N-linked glycan side on the head region of the HA and are fully resistant to inhibition by native SP-D. Importantly, we now show that D325A+R343V and D325S+R343V inhibited Cal09 H1N1 and related strains, and reduced uptake of Cal09 by epithelial cells. Inhibition of Cal09 was mediated by the lectin activity of the NCRDs. All known human pandemic strains have at least one glycan attachment on the top or side of the HA head, and our results indicate that they may be susceptible to inhibition by modified host defense lectins.
The Pseudomonas aeruginosa exoenzyme Y impairs endothelial cell proliferation and vascular repair following lung injury
Authors: Stevens Trevor C. , Ochoa Cristhiaan D. , Morrow K. Adam , Robson Matthew J. , Prasain Nutan , Zhou Chun , Alvarez Diego F. , Frank Dara W. , Balczon Ron , Stevens Troy
Published: April 2014

Exoenzyme Y (ExoY) is a Pseudomonas aeruginosa toxin that is introduced into host cells through the type 3 secretion system (T3SS). Once inside the host cell cytoplasm, ExoY generates cyclic nucleotides that cause tau phosphorylation and microtubule breakdown. Microtubule breakdown causes interendothelial cell gap formation and tissue edema. Although ExoY transiently induces interendothelial cell gap formation, it remains unclear whether ExoY prevents repair of the endothelial cell barrier. Here, we test the hypothesis that ExoY intoxication impairs recovery of the endothelial cell barrier following gap formation, decreasing migration, proliferation, and lung repair. Pulmonary microvascular endothelial cells (PMVECs) were infected with P. aeruginosa strains for 6 h, including one possessing an active ExoY (PA103 exoUexoT::Tc pUCPexoY; ExoY+), one with an inactive ExoY (PA103ΔexoUexoT::Tc pUCPexoYK81M; ExoYK81M), and one that lacks PcrV required for a functional T3SS (ΔPcrV). ExoY+ induced interendothelial cell gaps, whereas ExoYK81M and ΔPcrV did not promote gap formation. Following gap formation, bacteria were removed and endothelial cell repair was examined. PMVECs were unable to repair gaps even 3–5 days after infection. Serum-stimulated growth was greatly diminished following ExoY intoxication. Intratracheal inoculation of ExoY+ and ExoYK81M caused severe pneumonia and acute lung injury. However, whereas the pulmonary endothelial cell barrier was functionally improved 1 wk following ExoYK81M infection, pulmonary endothelium was unable to restrict the hyperpermeability response to elevated hydrostatic pressure following ExoY+ infection. In conclusion, ExoY is an edema factor that chronically impairs endothelial cell barrier integrity following lung injury.