bluuebunny/crossref_metadata_2025 huggingface dataset:
The Cockroach Origin of the Termite Gut Microbiota: Patterns in Bacterial Community Structure Reflect Major Evolutionary Events
Authors: Dietrich Carsten , Köhler Tim , Brune Andreas
Published: February 2014

Termites digest wood and other lignocellulosic substrates with the help of their intestinal microbiota. While the functions of the symbionts in the digestive process are slowly emerging, the origin of the bacteria colonizing the hindgut bioreactor is entirely unknown. Recently, our group discovered numerous representatives of bacterial lineages specific to termite guts in a closely related omnivorous cockroach, but it remains unclear whether they derive from the microbiota of a common ancestor or were independently selected by the gut environment. Here, we studied the bacterial gut microbiota in 34 species of termites and cockroaches using pyrotag analysis of the 16S rRNA genes. Although the community structures differed greatly between the major host groups, with dramatic changes in the relative abundances of particular bacterial taxa, we found that the majority of sequence reads belonged to bacterial lineages that were shared among most host species. When mapped onto the host tree, the changes in community structure coincided with major events in termite evolution, such as acquisition and loss of cellulolytic protists and the ensuing dietary diversification. UniFrac analysis of the core microbiota of termites and cockroaches and construction of phylogenetic tree of individual genus level lineages revealed a general host signal, whereas the branching order often did not match the detailed phylogeny of the host. It remains unclear whether the lineages in question have been associated with the ancestral cockroach since the early Cretaceous (cospeciation) or are diet-specific lineages that were independently acquired from the environment (host selection).
Inactivation of Human Norovirus in Contaminated Oysters and Clams by High Hydrostatic Pressure
Authors: Ye Mu , Li Xinhui , Kingsley David H. , Jiang Xi , Chen Haiqiang
Published: February 2014

Human norovirus (NoV) is the most frequent causative agent of food-borne disease associated with shellfish consumption. In this study, the effect of high hydrostatic pressure (HHP) on inactivation of NoV was determined. Genogroup I.1 (GI.1) or genogroup II.4 (GII.4) NoV was inoculated into oyster homogenates and treated at 300 to 600 MPa at 25, 6, and 1°C for 5 min. After HHP, samples were treated with RNase and viral particles were extracted with porcine gastric mucin (PGM)-conjugated magnetic beads (PGM-MBs). Viral RNA was then quantified by real-time reverse transcription (RT)-PCR. Since PGM contains histo-blood group-like antigens, which can act as receptors for NoV, deficiency for binding to PGM is an indication of loss of infectivity of NoV. After binding to PGM-MBs, RT-PCR-detectable NoV RNA in oysters was reduced by 0.4 to >4 log
10
by HHP at 300 to 600 MPa. The GI.1 NoV was more resistant to HHP than the GII.4 NoV (
P
< 0.05). HHP at lower temperatures significantly enhanced the inactivation of NoV in oysters (
P
< 0.05). Pressure treatment was also conducted for clam homogenates. Treatment at 450 MPa at 1°C achieved a >4 log
10
reduction of GI.1 NoV in both oyster and clam homogenates. It is therefore concluded that HHP could be applied as a potential intervention for inactivating NoV in raw shellfish. The method of pretreatment of samples with RNase, extraction of viral particles using PGM-MB binding, and quantification of viral RNA using RT-PCR can be explored as a practical means of distinguishing between infectious and noninfectious NoV.
H
2
O
2
Production in Species of the Lactobacillus acidophilus Group: a Central Role for a Novel NADH-Dependent Flavin Reductase
Authors: Hertzberger Rosanne , Arents Jos , Dekker Henk L. , Pridmore R. David , Gysler Christof , Kleerebezem Michiel , de Mattos M. Joost Teixeira
Published: February 2014

Hydrogen peroxide production is a well-known trait of many bacterial species associated with the human body. In the presence of oxygen, the probiotic lactic acid bacterium
Lactobacillus johnsonii
NCC 533 excretes up to 1 mM H
2
O
2
, inducing growth stagnation and cell death. Disruption of genes commonly assumed to be involved in H
2
O
2
production (e.g., pyruvate oxidase, NADH oxidase, and lactate oxidase) did not affect this. Here we describe the purification of a novel NADH-dependent flavin reductase encoded by two highly similar genes (
LJ
_
0548
and
LJ
_
0549
) that are conserved in lactobacilli belonging to the
Lactobacillus acidophilus
group. The genes are predicted to encode two 20-kDa proteins containing flavin mononucleotide (FMN) reductase conserved domains. Reductase activity requires FMN, flavin adenine dinucleotide (FAD), or riboflavin and is specific for NADH and not NADPH. The

K
m

for FMN is 30 ± 8 μM, in accordance with its proposed
in vivo
role in H
2
O
2
production. Deletion of the encoding genes in
L. johnsonii
led to a 40-fold reduction of hydrogen peroxide formation. H
2
O
2
production in this mutant could only be restored by in
trans
complementation of both genes. Our work identifies a novel, conserved NADH-dependent flavin reductase that is prominently involved in H
2
O
2
production in
L. johnsonii
.
In-Feed Use of Heavy Metal Micronutrients in U.S. Swine Production Systems and Its Role in Persistence of Multidrug-Resistant Salmonellae
Authors: Medardus Julius J. , Molla Bayleyegn Z. , Nicol Matthew , Morrow W. Morgan , Rajala-Schultz Paivi J. , Kazwala Rudovick , Gebreyes Wondwossen A.
Published: February 2014

The study aimed to characterize the role of heavy metal micronutrients in swine feed in emergence of heavy-metal-tolerant and multidrug-resistantSalmonellaorganisms. We conducted a longitudinal study in 36 swine barns over a 2-year period. The feed and fecal levels of Cu2+and Zn2+were measured.Salmonellawas isolated at early and late finishing. MICs of copper sulfate and zinc chloride were measured using agar dilution. Antimicrobial susceptibility was tested using the Kirby-Bauer method, and 283 isolates were serotyped. We amplifiedpcoAandczcDgenes that encode Cu2+and Zn2+tolerance, respectively. Of the 283 isolates, 113 (48%) showed Cu2+tolerance at 24 mM and 164 (58%) showed Zn2+tolerance at 8 mM. In multivariate analysis, serotype and source of isolates were significantly associated with Cu2+tolerance (P< 0.001). Fecal isolates were more likely to be Cu2+tolerant than those of feed origin (odds ratio [OR], 27.0; 95% confidence interval [CI], 2.8 to 250;P= 0.0042) or environmental origin (OR, 5.8), implying the significance of gastrointestinal selective pressure.Salmonella entericaserotypes Typhimurium and Heidelberg, highly significant for public health, had higher odds of having >20 mM MICs of Cu2+than did “other” serotypes. More than 60% ofSalmonellaisolates with resistance type (R-type) AmStTeKm (32 of 53) carriedpcoA; only 5% with R-type AmClStSuTe carried this gene.czcDgene carriage was significantly associated with a higher Zn2+MIC (P< 0.05). The odds of having a high Zn2+MIC (≥8 mM) were 14.66 times higher in isolates with R-type AmClStSuTe than in those with R-type AmStTeKm (P< 0.05). The findings demonstrate strong association between heavy metal tolerance and antimicrobial resistance, particularly amongSalmonellaserotypes important in public health.
Two-Component Histidine Phosphotransfer Protein Ypd1 Is Not Essential for Viability in Candida albicans
Authors: Mavrianos John , Desai Chirayu , Chauhan Neeraj
Published: February 2014

Prokaryotes and lower eukaryotes, such as yeasts, utilize two-component signal transduction pathways to adapt cells to environmental stress and to regulate the expression of genes associated with virulence. One of the central proteins in this type of signaling mechanism is the phosphohistidine intermediate protein Ypd1. Ypd1 is reported to be essential for viability in the model yeast
Saccharomyces cerevisiae
. We present data here showing that this is not the case for
Candida albicans
. Disruption of
YPD1
causes cells to flocculate and filament constitutively under conditions that favor growth in yeast form. To determine the function of Ypd1 in the Hog1 mitogen-activated protein kinase (MAPK) pathway, we measured phosphorylation of Hog1 MAPK in
ypd1
Δ/Δ and wild-type strains of
C. albicans
. Constitutive phosphorylation of Hog1 was observed in the
ypd1
Δ/Δ strain compared to the wild-type strain. Furthermore, fluorescence microscopy revealed that green fluorescent protein (GFP)-tagged Ypd1 is localized to both the nucleus and the cytoplasm. The subcellular segregation of GFP-tagged Ypd1 hints at an important role(s) of Ypd1 in regulation of Ssk1 (cytosolic) and Skn7 (nuclear) response regulator proteins via phosphorylation in
C. albicans
. Overall, our findings have profound implications for a mechanistic understanding of two-component signaling pathways in
C. albicans
, and perhaps in other pathogenic fungi.
The Acetylation Motif in AMP-Forming Acyl Coenzyme A Synthetases Contains Residues Critical for Acetylation and Recognition by the Protein Acetyltransferase Pat of Rhodopseudomonas palustris
Authors: Crosby Heidi A. , Escalante-Semerena Jorge C.
Published: February 2014

The AMP-forming acyl coenzyme A (acyl-CoA) synthetases are a large class of enzymes found in both anabolic and catabolic pathways that activate fatty acids to acyl-CoA molecules. The protein acetyltransferase (Pat) from
Rhodopseudomonas palustris
(
Rp
Pat) inactivates AMP-forming acyl-CoA synthetases by acetylating the ε-amino group of a conserved, catalytic lysine residue. In all of the previously described
Rp
Pat substrates, this lysine residue is located within a PX
4
GK motif that has been proposed to be a recognition motif for
Rp
Pat. Here, we report five new substrates for
Rp
Pat, all of which are also AMP-forming acyl-CoA synthetases. This finding supports the idea that Pat enzymes may have evolved to control the activity of this family of enzymes. Notably,
Rp
Pat did not acetylate the wild-type long-chain acyl-CoA synthetase B (
Rp
LcsB; formerly Rpa2714) enzyme of this bacterium. However, a single amino acid change two residues upstream of the acetylation site was sufficient to convert
Rp
LcsB into an
Rp
Pat substrate. The results of mutational and functional analyses of
Rp
LcsB and
Rp
PimA variants led us to propose PK/R
T
X
S/T/V/N
GKX
2
K/R as a substrate recognition motif. The underlined positions within this motif are particularly important for acetylation by
Rp
Pat. The first residue, threonine, is located 4 amino acids upstream of the acetylation site. The second residue can be S/T/V/N and is located two positions upstream of the acetylation site. Analysis of published crystal structures suggests that the side chains of these two residues are very close to the acetylated lysine residue, indicating that they may directly interact with
Rp
Pat.
Roles of Cyclic Di-GMP and the Gac System in Transcriptional Control of the Genes Coding for the Pseudomonas putida Adhesins LapA and LapF
Authors: Martínez-Gil Marta , Ramos-González María Isabel , Espinosa-Urgel Manuel
Published: February 2014

LapA and LapF are large extracellular proteins that play a relevant role in biofilm formation by
Pseudomonas putida
. Current evidence favors a sequential model in which LapA is first required for the initial adhesion of individual bacteria to a surface, while LapF participates in later stages of biofilm development. In agreement with this model,
lapF
transcription was previously shown to take place at late times of growth and to respond to the stationary-phase sigma factor RpoS. We have now analyzed the transcription pattern of
lapA
and other regulatory elements that influence expression of both genes. The
lapA
promoter shows a transient peak of activation early during growth, with a second increase in stationary phase that is independent of RpoS. The same pattern is observed in biofilms although expression is not uniform in the population. Both
lapA
and
lapF
are under the control of the two-component regulatory system GacS/GacA, and their transcription also responds to the intracellular levels of the second messenger cyclic diguanylate (c-di-GMP), although in surprisingly reverse ways. Whereas expression from the
lapA
promoter increases with high levels of c-di-GMP, the opposite is true for
lapF
. The transcriptional regulator FleQ is required for the modulation of
lapA
expression by c-di-GMP but has a minor influence on
lapF
. This work represents a further step in our understanding of the regulatory interactions controlling biofilm formation in
P. putida
.
Natural Competence and the Evolution of DNA Uptake Specificity
Authors: Mell Joshua Chang , Redfield Rosemary J.
Published: February 2014

Many bacteria are naturally competent, able to actively transport environmental DNA fragments across their cell envelope and into their cytoplasm. Because incoming DNA fragments can recombine with and replace homologous segments of the chromosome, competence provides cells with a potent mechanism of horizontal gene transfer as well as access to the nutrients in extracellular DNA. This review starts with an introductory overview of competence and continues with a detailed consideration of the DNA uptake specificity of competent proteobacteria in the
Pasteurellaceae
and
Neisseriaceae
. Species in these distantly related families exhibit strong preferences for genomic DNA from close relatives, a self-specificity arising from the combined effects of biases in the uptake machinery and genomic overrepresentation of the sequences this machinery prefers. Other competent species tested lack obvious uptake bias or uptake sequences, suggesting that strong convergent evolutionary forces have acted on these two families. Recent results show that uptake sequences have multiple “dialects,” with clades within each family preferring distinct sequence variants and having corresponding variants enriched in their genomes. Although the genomic consensus uptake sequences are 12 and 29 to 34 bp, uptake assays have found that only central cores of 3 to 4 bp, conserved across dialects, are crucial for uptake. The other bases, which differ between dialects, make weaker individual contributions but have important cooperative interactions. Together, these results make predictions about the mechanism of DNA uptake across the outer membrane, supporting a model for the evolutionary accumulation and stability of uptake sequences and suggesting that uptake biases may be more widespread than currently thought.
Functional Analysis of the Accessory Protein TapA in Bacillus subtilis Amyloid Fiber Assembly
Authors: Romero Diego , Vlamakis Hera , Losick Richard , Kolter Roberto
Published: February 2014

Bacillus subtilis
biofilm formation relies on the assembly of a fibrous scaffold formed by the protein TasA. TasA polymerizes into highly stable fibers with biochemical and morphological features of functional amyloids. Previously, we showed that assembly of TasA fibers requires the auxiliary protein TapA. In this study, we investigated the roles of TapA sequences from the C-terminal and N-terminal ends and TapA cysteine residues in its ability to promote the assembly of TasA amyloid-like fibers. We found that the cysteine residues are not essential for the formation of TasA fibers, as their replacement by alanine residues resulted in only minor defects in biofilm formation. Mutating sequences in the C-terminal half had no effect on biofilm formation. However, we identified a sequence of 8 amino acids in the N terminus that is key for TasA fiber formation. Strains expressing TapA lacking these 8 residues were completely defective in biofilm formation. In addition, this TapA mutant protein exhibited a dominant negative effect on TasA fiber formation. Even in the presence of wild-type TapA, the mutant protein inhibited fiber assembly
in vitro
and delayed biofilm formation
in vivo
. We propose that this 8-residue sequence is crucial for the formation of amyloid-like fibers on the cell surface, perhaps by mediating the interaction between TapA or TapA and TasA molecules.
Comparative Functional Genomics of Lactobacillus spp. Reveals Possible Mechanisms for Specialization of Vaginal Lactobacilli to Their Environment
Authors: Mendes-Soares Helena , Suzuki Haruo , Hickey Roxana J. , Forney Larry J.
Published: February 2014

Lactobacilli are found in a wide variety of habitats. Four species,
Lactobacillus crispatus
,
L. gasseri
,
L. iners
, and
L. jensenii
, are common and abundant in the human vagina and absent from other habitats. These may be adapted to the vagina and possess characteristics enabling them to thrive in that environment. Furthermore, stable codominance of multiple
Lactobacillus
species in a single community is infrequently observed. Thus, it is possible that individual vaginal
Lactobacillus
species possess unique characteristics that confer to them host-specific competitive advantages. We performed comparative functional genomic analyses of representatives of 25 species of
Lactobacillus
, searching for habitat-specific traits in the genomes of the vaginal lactobacilli. We found that the genomes of the vaginal species were significantly smaller and had significantly lower GC content than those of the nonvaginal species. No protein families were found to be specific to the vaginal species analyzed, but some were either over- or underrepresented relative to nonvaginal species. We also found that within the vaginal species, each genome coded for species-specific protein families. Our results suggest that even though the vaginal species show no general signatures of adaptation to the vaginal environment, each species has specific and perhaps unique ways of interacting with its environment, be it the host or other microbes in the community. These findings will serve as a foundation for further exploring the role of lactobacilli in the ecological dynamics of vaginal microbial communities and their ultimate impact on host health.
Germination of Spores of Bacillus Species: What We Know and Do Not Know
Authors: Setlow Peter
Published: February 2014

Spores of
Bacillus
species can remain in their dormant and resistant states for years, but exposure to agents such as specific nutrients can cause spores' return to life within minutes in the process of germination. This process requires a number of spore-specific proteins, most of which are in or associated with the inner spore membrane (IM). These proteins include the (i) germinant receptors (GRs) that respond to nutrient germinants, (ii) GerD protein, which is essential for GR-dependent germination, (iii) SpoVA proteins that form a channel in spores' IM through which the spore core's huge depot of dipicolinic acid is released during germination, and (iv) cortex-lytic enzymes (CLEs) that degrade the large peptidoglycan cortex layer, allowing the spore core to take up much water and swell, thus completing spore germination. While much has been learned about nutrient germination, major questions remain unanswered, including the following. (i) How do nutrient germinants penetrate through spores' outer layers to access GRs in the IM? (ii) What happens during the highly variable and often long lag period between the exposure of spores to nutrient germinants and the commitment of spores to germinate? (iii) What do GRs and GerD do, and how do these proteins interact? (iv) What is the structure of the SpoVA channel in spores' IM, and how is this channel gated? (v) What is the precise state of the spore IM, which has a number of novel properties even though its lipid composition is very similar to that of growing cells? (vi) How is CLE activity regulated such that these enzymes act only when germination has been initiated? (vii) And finally, how does the germination of spores of clostridia compare with that of spores of bacilli?
RNase E Affects the Expression of the Acyl-Homoserine Lactone Synthase Gene
sinI
in Sinorhizobium meliloti
Authors: Baumgardt Kathrin , Charoenpanich Pornsri , McIntosh Matthew , Schikora Adam , Stein Elke , Thalmann Sebastian , Kogel Karl-Heinz , Klug Gabriele , Becker Anke , Evguenieva-Hackenberg Elena
Published: February 2014

Quorum sensing of
Sinorhizobium meliloti
relies on
N
-acyl-homoserine lactones (AHLs) as autoinducers. AHL production increases at high population density, and this depends on the AHL synthase SinI and two transcriptional regulators, SinR and ExpR. Our study demonstrates that ectopic expression of the gene
rne
, coding for RNase E, an endoribonuclease that is probably essential for growth, prevents the accumulation of AHLs at detectable levels. The ectopic
rne
expression led to a higher level of
rne
mRNA and a lower level of
sinI
mRNA independently of the presence of ExpR, the AHL receptor, and AHLs. In line with this, IPTG (isopropyl-β-
d
-thiogalactopyranoside)-induced overexpression of
rne
resulted in a shorter half-life of
sinI
mRNA and a strong reduction of AHL accumulation. Moreover, using translational
sinI-egfp
fusions, we found that
sinI
expression is specifically decreased upon induced overexpression of
rne
, independently of the presence of the global posttranscriptional regulator Hfq. The 28-nucleotide 5′ untranslated region (UTR) of
sinI
mRNA was sufficient for this effect. Random amplification of 5′ cDNA ends (5′-RACE) analyses revealed a potential RNase E cleavage site at position +24 between the Shine-Dalgarno site and the translation start site. We postulate therefore that RNase E-dependent degradation
of sinI
mRNA from the 5′ end is one of the steps mediating a high turnover of
sinI
mRNA, which allows the Sin quorum-sensing system to respond rapidly to changes in transcriptional control of AHL production.
Mechanics of turning and jumping and skier speed are associated with injury risk in men's World Cup alpine skiing: a comparison between the competition disciplines
Authors: Gilgien Matthias , Spörri Jörg , Kröll Josef , Crivelli Philip , Müller Erich
Published: February 2014

Background/aim

In alpine ski racing, there is limited information about skiers’ mechanical characteristics and their relation to injury risk, in particular for World Cup (WC) competitions. Hence, current findings from epidemiological and qualitative research cannot be linked to skiers’ mechanics. This study was undertaken to investigate whether recently reported differences in numbers of injuries per 1000 runs for competition disciplines can be explained by differences in the skiers’ mechanics.

Methods

During seven giant slalom, four super-G and five downhill WC competitions, mechanical characteristics of a forerunner were captured using differential global navigation satellite technology and a precise terrain surface model. Finally, the discipline-specific skiers’ mechanics were compared with the respective number of injuries per hour skiing.

Results

While the number of injuries per hour skiing was approximately equal for all disciplines, kinetic energy, impulse, run time, turn radius and turn speed were significantly different and increased from giant slalom to super-G and downhill. Turn ground reaction forces were largest for giant slalom, followed by super-G and downhill. The number of jumps was doubled from super-G to downhill.

Conclusions

Associating the number of injuries per hour in WC skiing with skiers’ mechanical characteristics, injuries in super-G and downhill seem to be related to increased speed and jumps, while injuries in giant slalom may be related to high loads in turning. The reported differences in the number of injuries per 1000 runs might be explained by a bias in total exposure time per run and thus potentially by emerged fatigue.
Hospital-based surveillance of rotavirus gastroenteritis among children under 5 years of age in the Republic of Ivory Coast: a cross-sectional study
Authors: Akoua-Koffi Chantal , Asse Kouadio Vincent , Yao Atteby Jean Jacques
Published: February 2014

Objectives

To estimate the proportion of rotavirus gastroenteritis (RVGE) among children aged less than 5 years who had been diagnosed with acute gastroenteritis (AGE) and admitted to hospitals and emergency rooms (ERs). The seasonal distribution of RVGE and most prevalent rotavirus (RV) strains was also assessed.

Design

A cross-sectional hospital-based surveillance study.

Setting

5 reference paediatric hospitals across Abidjan.

Participants

Children aged less than 5 years, who were hospitalised/visiting ERs for WHO-defined AGE, were enrolled. Written informed consent was obtained from parents/guardians before enrolment. Children who acquired nosocomial infection were excluded from the study.

Primary and secondary outcome measures

The proportion of RVGE among AGE hospitalisations and ER visits was expressed with 95% exact CI. Stool samples were collected from all enrolled children and were tested for the presence of RV using an enzyme immunoassay. RV-positive samples were serotyped using reverse transcriptase-PCR.

Results

Of 357 enrolled children (mean age 13.6±11.14 months), 332 were included in the final analyses; 56.3% (187/332) were hospitalised and 43.7% (145/332) were admitted to ERs. The proportion of RVGE hospitalisations and ER visits among all AGE cases was 30.1% (95% CI 23.6% to 37.3%) and 26.9% (95% CI 19.9% to 34.9%), respectively. Ninety-five children (28.6%) were RV positive; the highest number of RVGE cases was observed in children aged 6–11 months. The number of GE cases peaked in July and August 2008; the highest percentage of RV-positive cases was observed in January 2008. G1P[8] wild-type and G8P[6] were the most commonly detected strains.

Conclusions

RVGE causes substantial morbidity among children under 5 years of age and remains a health concern in the Republic of Ivory Coast, where implementation of prevention strategies such as vaccination might help to reduce disease burden.
Journey to vaccination: a protocol for a multinational qualitative study
Authors: Wheelock Ana , Miraldo Marisa , Parand Anam , Vincent Charles , Sevdalis Nick
Published: February 2014

Introduction

In the past two decades, childhood vaccination coverage has increased dramatically, averting an estimated 2–3 million deaths per year. Adult vaccination coverage, however, remains inconsistently recorded and substandard. Although structural barriers are known to limit coverage, social and psychological factors can also affect vaccine uptake. Previous qualitative studies have explored beliefs, attitudes and preferences associated with seasonal influenza (flu) vaccination uptake, yet little research has investigated how participants’ context and experiences influence their vaccination decision-making process over time. This paper aims to provide a detailed account of a mixed methods approach designed to understand the wider constellation of social and psychological factors likely to influence adult vaccination decisions, as well as the context in which these decisions take place, in the USA, the UK, France, India, China and Brazil.

Methods and analysis

We employ a combination of qualitative interviewing approaches to reach a comprehensive understanding of the factors influencing vaccination decisions, specifically seasonal flu and tetanus. To elicit these factors, we developed thejourney to vaccination, a new qualitative approach anchored on theheuristics and biasestradition and thecustomer journey mappingapproach. A purposive sampling strategy is used to select participants who represent a range of key sociodemographic characteristics. Thematic analysis will be used to analyse the data. Typical journeys to vaccination will be proposed.

Ethics and dissemination

Vaccination uptake is significantly influenced by social and psychological factors, some of which are under-reported and poorly understood. This research will provide a deeper understanding of the barriers and drivers to adult vaccination. Our findings will be published in relevant peer-reviewed journals and presented at academic conferences. They will also be presented as practical recommendations at policy and industry meetings and healthcare professionals’ forums. This research was approved by relevant local ethics committees.
A randomised controlled trial of a consumer-focused e-health strategy for cardiovascular risk management in primary care: the Consumer Navigation of Electronic Cardiovascular Tools (CONNECT) study protocol
Authors: Redfern Julie , Usherwood T , Harris M F , Rodgers A , Hayman N , Panaretto K , Chow C , Lau A Y S , Neubeck L , Coorey G , Hersch F , Heeley E , Patel A , Jan S , Zwar N , Peiris D
Published: February 2014

Introduction

Fewer than half of all people at highest risk of a cardiovascular event are receiving and adhering to best practice recommendations to lower their risk. In this project, we examine the role of an e-health-assisted consumer-focused strategy as a means of overcoming these gaps between evidence and practice. Consumer Navigation of Electronic Cardiovascular Tools (CONNECT) aims to test whether a consumer-focused e-health strategy provided to Aboriginal and Torres Strait Islander and non-indigenous adults, recruited through primary care, at moderate-to-high risk of a cardiovascular disease event will improve risk factor control when compared with usual care.

Methods and analysis

Randomised controlled trial of 2000 participants with an average of 18 months of follow-up to evaluate the effectiveness of an integrated consumer-directed e-health portal on cardiovascular risk compared with usual care in patients with cardiovascular disease or who are at moderate-to-high cardiovascular disease risk. The trial will be augmented by formal economic and process evaluations to assess acceptability, equity and cost-effectiveness of the intervention. The intervention group will participate in a consumer-directed e-health strategy for cardiovascular risk management. The programme is electronically integrated with the primary care provider's software and will include interactive smart phone and Internet platforms. The primary outcome is a composite endpoint of the proportion of people meeting the Australian guideline-recommended blood pressure (BP) and cholesterol targets. Secondary outcomes include change in mean BP and fasting cholesterol levels, proportion meeting BP and cholesterol targets separately, self-efficacy, health literacy, self-reported point prevalence abstinence in smoking, body mass index and waist circumference, self-reported physical activity and self-reported medication adherence.

Ethics and dissemination

Primary ethics approval was received from the University of Sydney Human Research Ethics Committee and the Aboriginal Health and Medical Research Council. Results will be disseminated via the usual scientific forums including peer-reviewed publications and presentations at international conferences

Clinical Trials registration number

ACTRN12613000715774.
Infrequent and incomplete registration of test accuracy studies: analysis of recent study reports
Authors: Korevaar Daniël A , Bossuyt Patrick M M , Hooft Lotty
Published: February 2014

Objectives

To identify the proportion of articles reporting on test accuracy for which the corresponding study had been registered.

Design

Analysis of a consecutive sample of published study reports.

Participants

PubMed was searched for publications in journals with an impact factor of 5 or higher in May and June 2012. Articles were included if they reported on original studies evaluating the accuracy of one or more diagnostic or prognostic tests or markers against a clinical reference standard in humans.

Primary and secondary outcome measures

Primary outcome was registration of the reported test accuracy study. We additionally explored study characteristics associated with registration.

Results

We found 1941 references; 351 study reports fulfilled the inclusion criteria, of which 52 studies (15%) had been registered. Of these, 27 (52%) provided a registration number in the publication, and 12 (23%) provided a reference to the publication in the registry. Registration rates were similar for studies on diagnostic versus those on prognostic tests, and among studies on imaging tests versus those on laboratory techniques. Studies reporting some form of industry involvement were more often registered (33%) than studies reporting another source of funding (11%), and studies without a (reported) source of (external) funding (9%; p<0.001). Of the registered studies, 8 (15%) had been registered after completion, 14 were registered before initiation (27%) and 30 (58%) between initiation and completion. Only 16 (31%; 5% of the total sample) had registered the published primary outcome measures before completion.

Conclusions

Few test accuracy studies published in higher impact journals are registered. Only 1 in 22 of such studies register their primary outcomes before study completion. Owing to the reasons for registering studies that investigate the cause-and-effect relationship between health-related interventions and health outcomes also apply to test accuracy studies, prospective study registration of these studies should be further promoted among investigators and journal editors.
The preventive effects of low-dose enteric-coated aspirin tablets on the development of colorectal tumours in Asian patients: a randomised trial
Authors: Ishikawa Hideki , Mutoh Michihiro , Suzuki Sadao , Tokudome Shinkan , Saida Yoshihisa , Abe Takashi , Okamura Shozo , Tajika Masahiro , Joh Takashi , Tanaka Shinji , Kudo Shin-ei , Matsuda Takahisa , Iimuro Masaki , Yukawa Tomomi , Takayama Tetsuji , Sato Yasushi , Lee Kyowon , Kitamura Shinji , Mizuno Motowo , Sano Yasushi , Gondo Nobuhisa , Sugimoto Kenji , Kusunoki Masato , Goto Chiho , Matsuura Nariaki , Sakai Toshiyuki , Wakabayashi Keiji
Published: February 2014

Objective

To evaluate the influence of low-dose, enteric-coated aspirin tablets (100 mg/day for 2 years) on colorectal tumour recurrence in Asian patients with single/multiple colorectal tumours excised by endoscopy.

Design

A double-blinded, randomised, placebo-controlled multicentre clinical trial was conducted.

Participants

311 subjects with single/multiple colorectal adenomas and adenocarcinomas excised by endoscopy were enrolled in the study (152 patients in the aspirin group and 159 patients in the placebo group). Enrolment began at the hospitals (n=19) in 2007 and was completed in 2009.

Results

The subjects treated with aspirin displayed reduced colorectal tumourigenesis and primary endpoints with an adjusted OR of 0.60 (95% CI 0.36 to 0.98) compared with the subjects in the placebo group. Subgroup analysis revealed that subjects who were non-smokers, defined as those who had smoked in the past or who had never smoked, had a marked reduction in the number of recurrent tumours in the aspirin-treated group. The adjusted OR for aspirin treatment in non-smokers was 0.37 (CI 0.21 to 0.68, p<0.05). Interestingly, the use of aspirin in smokers resulted in an increased risk, with an OR of 3.44. In addition, no severe adverse effects were observed in either group.

Conclusions

Low-dose, enteric-coated aspirin tablets reduced colorectal tumour recurrence in an Asian population. The results are consistent with those obtained from other randomised controlled trials in Western countries.

The clinical trial registry website and the clinical trial number

http://www.umin.ac.jp(number UMIN000000697).
Cell lineage distribution atlas of the human stomach reveals heterogeneous gland populations in the gastric antrum
Authors: Choi Eunyoung , Roland Joseph T , Barlow Brittney J , O'Neal Ryan , Rich Amy E , Nam Ki Taek , Shi Chanjuan , Goldenring James R
Published: February 2014

Objective

The glands of the stomach body and antral mucosa contain a complex compendium of cell lineages. In lower mammals, the distribution of oxyntic glands and antral glands define the anatomical regions within the stomach. We examined in detail the distribution of the full range of cell lineages within the human stomach.

Design

We determined the distribution of gastric gland cell lineages with specific immunocytochemical markers in entire stomach specimens from three non-obese organ donors.

Results

The anatomical body and antrum of the human stomach were defined by the presence of ghrelin and gastrin cells, respectively. Concentrations of somatostatin cells were observed in the proximal stomach. Parietal cells were seen in all glands of the body of the stomach as well as in over 50% of antral glands. MIST1 expressing chief cells were predominantly observed in the body although individual glands of the antrum also showed MIST1 expressing chief cells. While classically described antral glands were observed with gastrin cells and deep antral mucous cells without any parietal cells, we also observed a substantial population of mixed type glands containing both parietal cells and G cells throughout the antrum.

Conclusions

Enteroendocrine cells show distinct patterns of localisation in the human stomach. The existence of antral glands with mixed cell lineages indicates that human antral glands may be functionally chimeric with glands assembled from multiple distinct stem cell populations.
Release kinetics of early ischaemic biomarkers in a clinical model of acute myocardial infarction
Authors: Liebetrau Christoph , Nef Holger M , Dörr Oliver , Gaede Luise , Hoffmann Jedrzej , Hahnel Astrid , Rolf Andreas , Troidl Christian , Lackner Karl J , Keller Till , Hamm Christian W , Möllmann Helge
Published: February 2014

Objective

To determine the release kinetics of different biomarkers with potential as novel early ischaemic biomarkers in patients with acute coronary syndrome (ACS); it is difficult to establish the detailed release kinetics in patients with acute myocardial infarction (AMI).

Methods

We analysed the release kinetics of soluble fms-like tyrosine kinase (sFlt-1), ischaemia modified albumin (IMA), and heart-type fatty acid binding protein (hFABP) in patients with hypertrophic obstructive cardiomyopathy who were undergoing transcoronary ablation of septal hypertrophy (TASH), a procedure mimicking AMI. Consecutive patients (n=21) undergoing TASH were included. Blood samples were collected before TASH and 15, 30, 45, 60, 75, 90, and 105 min and 2, 4, 8, and 24 h after TASH. sFlt-1 and hFABP were quantified in serum, and IMA was quantified in plasma using immunoassays.

Results

sFLT-1 and hFABP increased significantly 15 min after induction of AMI vs baseline as follows: sFlt-1, 3657.5 ng/L (IQR 2302.3–4475.0) vs 76.0 ng/L (IQR 71.2–88.8) (p<0.001); hFABP, 9.0 ng/mL (IQR 7.0–15.4) vs 4.6 ng/mL (IQR 3.4–7.1) (p<0.001). sFlt-1 demonstrated a continuous decrease after the 15th min. hFABP showed a continuous increase until the 8th hour with a decline afterwards. The IMA concentrations increased significantly 30 min after induction of AMI vs baseline, with values of 26.0 U/mL (IQR 21.8–38.6) vs 15.6 U/mL (IQR 10.1–24.7) (p=0.02), and then decreased after 75 min.

Conclusions

sFlt-1 and hFABP increased very early after induction of myocardial ischaemia, showing different release kinetics. The additional information provided by these findings is helpful for developing their potential combined use with cardiac troponins in patients with suspected AMI.
Practical techniques for reducing radiation exposure during cerebral angiography procedures
Authors: Pearl Monica S , Torok Collin , Wang Jiangxia , Wyse Emily , Mahesh Mahadevappa , Gailloud Philippe
Published: February 2014

Purpose

DSA remains the gold standard imaging method for the evaluation of many cerebrovascular disorders, in particular cerebral aneurysms and vascular malformations. The purpose of this study was to demonstrate the effect of modifying DSA frame rate, fluoroscopic and roadmap pulse rates, and flat panel detector (FPD) position on the radiation dose delivered during routine views for a cerebral angiogram in a phantom model.

Materials and methods

Adult skull and abdomen/pelvis anthropomorphic phantoms were used to compare the radiation dose metrics Ka,r (in mGy), PKA (in μGym2), and fluoroscopy time (in minutes) after modification of fluoroscopic pulses per second (p/s), DSA frames per second (f/s), and FPD position and collimation in three components of a cerebral angiogram: (1) femoral artery access, (2) roadmap guidance, and (3) biplane cerebral DSA.

Results

For femoral artery access, DSA protocols resulted in significantly higher doses than those utilizing fluoroscopy alone (p=0.007). Roadmaps using 3 p/s or 4 p/s delivered significantly less dose than higher pulse rates (p=0.008). The ranges of delivered doses for biplane cerebral DSA were 347.3–1188.5 mGy and 3914.54–9518.78 μGym2. The lowest radiation doses were generated by the variable frame rate DSA protocols.

Conclusions

Replacing femoral arterial access evaluations by DSA with fluoroscopy, utilizing lower pulse rates during fluoroscopy and roadmap guidance, and choosing variable frame rates for DSA are simple techniques that may be considered by operators in their clinical practices to lower radiation dose during cerebral angiography procedures.
Hybrid surgery for dural arteriovenous fistula in the neurosurgical hybrid operating suite
Authors: Shen Shih-Chieh , Tsuei Yuang-Seng , Chen Wen-Hsien , Shen Chiung-Chyi
Published: February 2014

Treatment of a dural arteriovenous fistula (AVF), which is difficult to access by either the surgical or endovascular approach, is challenging. A hybrid technique, combining a microsurgical approach and endovascular embolization, can provide less invasive management of dural AVFs in a modern neurosurgical hybrid operating suite. We present a case of intracerebral hemorrhage in the left cerebellum secondary to dural AVF, Cognard type IV with numerous tiny feeders from the ascending pharyngeal artery branches. No adequate arterial or venous route for endovascular embolization was found by neuroangiography. The hybrid technique, combining keyhole pterional craniotomy and embolization with n-butyl cyanoacrylate glue injection via direct cannulation of the periclival venous plexus, succeeded in obliterating the dural AVF. Intraoperative angiography showed successful embolization of the dural AVF without any complication. This report illustrates the usefulness of the neurosurgical hybrid operating suite for the treatment of difficult dural AVFs.
Physical and electrical characterization of TiO2 particles after high temperature processing and before and after ultraviolet irradiation
Authors: Molina J. , Zúñiga C. , Moreno M. , Calleja W. , Rosales P. , Ambrosio R. , De la Hidalga F.J. , Gutiérrez E. , Heredia A. , Bandala E.R. , Sánchez J.L.
Published: February 2014

In this work, rutile-phase TiO2 particles (r-TiO2, about 360 nm in size) are embedded within a silicon oxide matrix using different concentration ratios of r-TiO2 with respect to SiO2:H2O, so that suspensions of mixed TiO2:SiO2 oxides were obtained and analyzed. These TiO2:SiO2 suspensions were deposited on previously-cleaned crystalline silicon and quartz substrates so that thin films of TiO2:SiO2 were obtained. All films were then exposed to relatively high-temperature thermal treatments in nitrogen and different characterization techniques were used to determine their physical and electrical properties before and after ultraviolet (UV) irradiation. Before high thermal treatment, X-ray diffraction patterns show that the main diffraction peaks for the obtained TiO2:SiO2 films correspond to the crystalline phase of rutile-TiO2. Infrared analyses before and after thermal treatment show significant changes in the chemical bonding of the final films relative to the temperatures used during annealing. Also, UV–visible spectra provide a constant optical band gap for the films, independent of different TiO2 concentrations as expected. On the other hand, atomic-force microscopy measurements before and after UV irradiation show an appreciable difference in the grain size and surface morphology of the resulting TiO2:SiO2 oxides annealed at 1000 °C. Finally, photoelectrical I–V properties were obtained for all TiO2:SiO2 films by depositing ultrathin titanium stripes on top of the photoactive material and then, measuring the total current flowing through the metal electrode before and after UV irradiation. From these last measurements, a detectable increase in the I–V slope (lower resistance of the titanium stripe) is found for all samples during UV exposure, thus making this device to act as a simple photoresistor based on r-TiO2 particles.
Characteristics of karst springs in Aydıncık (Mersin, Turkey), based on recession curves and hydrochemical and isotopic parameters
Authors: Hatipoglu-Bagci Zubeyde , Sazan Mehmet Selim
Published: February 2014

We evaluate the hydrogeology of the area around the Soğuksu spring in the Aydıncık district, Mersin, Turkey, with a focus on the groundwater and aquifer systems. An increasing population and the expansion of agricultural land have resulted in increasing demand for water in the Aydıncık area. The Soğuksu spring, discharging from the Aydıncık aquifer, is the major source of water for Aydıncık and the surrounding area. To establish a sustainable water management plan for the region it is essential to determine the characteristics of the hydrogeological structure of the area. Water samples taken from the Soğuksu spring and other small springs in the Aydıncık area were analysed and the spring recession curve, hydrochemistry, stable isotopes (δ
18
O, δ
2
H), and tritium isotopes (
3
H) were used to characterize the aquifer. The master recession coefficient obtained from analysis of the spring discharge data for 1999–2012 indicates one exponential term. Analysis of the groundwater composition identified two groups, the Ca–HCO
3
and Ca–Mg–HCO
3
groups, based on the major cation and major anion in the sampled water, as expected from karstic carbonate aquifers. The stable isotopes demonstrate the predominance of low-elevation precipitation. High tritium concentrations between 2.32 and 4.25 TU suggest recent recharge of groundwater from rainfall.
Microbiotic signatures of the Anthropocene in marginal marine and freshwater palaeoenvironments
Authors: Wilkinson I. P. , Poirier C. , Head M. J. , Sayer C. D. , Tibby J.
Published: February 2014

The term ‘Anthropocene’ has been proposed to indicate a geological interval characterized by global anthropogenic environmental change. This paper attempts to recognize a method by which the Anthropocene can be defined micropalaeontologically. In order to do this, microfloras and microfaunas (diatoms, macrophytes, dinoflagellate cysts, foraminifera and ostracods) from nearshore waters through to paralic and freshwater aquatic milieux are considered, and biotic variability with an anthropogenic causation identified. Microbiotic change can be related to anthropogenically induced extinctions, pollution-related mutation, environmentally influenced assemblage variability, geochemistry of carapaces/tests, floral change related to lacustrine acidification, faunal and floral correlation to industrial and agricultural signatures and introduction of exotic species via shipping. The influence of humanity on a local scale can be recognized in assemblages as far back as 5000 years BP. However, widespread anthropogenic change took place in Europe and America, particularly in the nineteenth and twentieth centuries, although in Asia (e.g. Japan) it cannot be observed prior to the twentieth century. Profound and global biotic change began in the mid-twentieth century and, if the Anthropocene is to be defined in this way, then the period 1940–1945 might encompass the biotic base of the interval.