bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ethylene Glycol Metabolism in the Acetogen Acetobacterium woodii
Authors: Trifunović Dragan , Schuchmann Kai , Müller Volker 
Published: January 2016

ABSTRACT

The acetogenic bacterium
Acetobacterium woodii
is able to grow by the oxidation of diols, such as 1,2-propanediol, 2,3-butanediol, or ethylene glycol. Recent analyses demonstrated fundamentally different ways for oxidation of 1,2-propanediol and 2,3-butanediol. Here, we analyzed the metabolism of ethylene glycol. Our data demonstrate that ethylene glycol is dehydrated to acetaldehyde, which is then disproportionated to ethanol and acetyl coenzyme A (acetyl-CoA). The latter is further converted to acetate, and this pathway is coupled to ATP formation by substrate-level phosphorylation. Apparently, the product ethanol is in part further oxidized and the reducing equivalents are recycled by reduction of CO
2
to acetate in the Wood-Ljungdahl pathway. Biochemical data as well as the results of protein synthesis analysis are consistent with the hypothesis that the propane diol dehydratase (PduCDE) and CoA-dependent propionaldehyde dehydrogenase (PduP) proteins, encoded by the
pdu
gene cluster, also catalyze ethylene glycol dehydration to acetaldehyde and its CoA-dependent oxidation to acetyl-CoA. Moreover, genes encoding bacterial microcompartments as part of the
pdu
gene cluster are also expressed during growth on ethylene glycol, arguing for a dual function of the Pdu microcompartment system.

IMPORTANCE
Acetogenic bacteria are characterized by their ability to use CO
2
as a terminal electron acceptor by a specific pathway, the Wood-Ljungdahl pathway, enabling in most acetogens chemolithoautotrophic growth with H
2
and CO
2
. However, acetogens are very versatile and can use a wide variety of different substrates for growth. Here we report on the elucidation of the pathway for utilization of ethylene glycol by the model acetogen
Acetobacterium woodii
. This diol is degraded by dehydration to acetaldehyde followed by a disproportionation to acetate and ethanol. We present evidence that this pathway is catalyzed by the same enzyme system recently described for the utilization of 1,2-propanediol. The enzymes for ethylene glycol utilization seem to be encapsulated in protein compartments, known as bacterial microcompartments.
Gene Deletion by Fluorescence-Reported Allelic Exchange Mutagenesis in Chlamydia trachomatis
Authors: Mueller Konrad E. , Wolf Katerina , Fields Kenneth A. 
Published: January 2016

ABSTRACT

Although progress in
Chlamydia
genetics has been rapid, genomic modification has previously been limited to point mutations and group II intron insertions which truncate protein products. The bacterium has thus far been intractable to gene deletion or more-complex genomic integrations such as allelic exchange. Herein, we present a novel suicide vector dependent on inducible expression of a chlamydial gene that renders
Chlamydia trachomatis
fully genetically tractable and permits rapid reverse genetics by fluorescence-reported allelic exchange mutagenesis (FRAEM). We describe the first available system of targeting chlamydial genes for deletion or allelic exchange as well as curing plasmids from
C. trachomatis
serovar L2. Furthermore, this approach permits the monitoring of mutagenesis by fluorescence microscopy without disturbing bacterial growth, a significant asset when manipulating obligate intracellular organisms. As proof of principle,
trpA
was successfully deleted and replaced with a sequence encoding both green fluorescent protein (GFP) and β-lactamase. The
trpA
-deficient strain was unable to grow in indole-containing medium, and this phenotype was reversed by complementation with
trpA
expressed in
trans
. To assess reproducibility at alternate sites, FRAEM was repeated for genes encoding type III secretion effectors CTL0063, CTL0064, and CTL0065. In all four cases, stable mutants were recovered one passage after the observation of transformants, and allelic exchange was limited to the specific target gene, as confirmed by whole-genome sequencing. Deleted sequences were not detected by quantitative real-time PCR (qPCR) from isogenic mutant populations. We demonstrate that utilization of the chlamydial suicide vector with FRAEM renders
C. trachomatis
highly amenable to versatile and efficient genetic manipulation.

IMPORTANCE
The obligate intracellular nature of a variety of infectious bacteria presents a significant obstacle to the development of molecular genetic tools for dissecting pathogenicity. Although progress in chlamydial genetics has been rapid, genomic modification has previously been limited to point mutations and group II intron insertions which truncate protein products. The bacterium has thus far been intractable to gene deletion or more-complex genomic integrations such as allelic exchange. Here, we present a novel suicide vector dependent on inducible expression of a chlamydial gene that renders
Chlamydia trachomatis
fully genetically tractable and permits rapid reverse genetics by fluorescence-reported allelic exchange mutagenesis (FRAEM). We describe the first available system of targeting chlamydial genes for deletion or allelic exchange as well as curing plasmids from
C. trachomatis
L2. Furthermore, this approach permits monitoring of mutagenesis by fluorescence microscopy without disturbing bacterial growth, a significant asset when manipulating obligate intracellular organisms.
Consideration of Epstein-Barr Virus-Encoded Noncoding RNAs EBER1 and EBER2 as a Functional Backup of Viral Oncoprotein Latent Membrane Protein 1
Authors: Herbert Kristina M. , Pimienta Genaro 
Published: January 2016

ABSTRACT

The Epstein-Barr virus (EBV)-encoded noncoding RNAs EBER1 and EBER2 are highly abundant through all four latency stages of EBV infection (III-II-I-0) and have been associated with an oncogenic phenotype when expressed in cell lines cultured
in vitro
.
In vivo
, EBV-infected B cells derived from freshly isolated lymphocytes show that EBER1/2 deletion does not impair viral latency. Based on published quantitative proteomics data from BJAB cells expressing EBER1 and EBER2, we propose that the EBERs, through their activation of AKT in a B-cell-specific manner, are a functionally redundant backup of latent membrane protein 1 (LMP1)—an essential oncoprotein in EBV-associated malignancies, with a main role in AKT activation. Our proposed model may explain the lack of effect on viral latency establishment in EBER-minus EBV infection.
The Stealthy Superbug: the Role of Asymptomatic Enteric Carriage in Maintaining a Long-Term Hospital Outbreak of ST228 Methicillin-Resistant Staphylococcus aureus
Authors: Senn Laurence , Clerc Olivier , Zanetti Giorgio , Basset Patrick , Prod’hom Guy , Gordon Nicola C. , Sheppard Anna E. , Crook Derrick W. , James Richard , Thorpe Harry A. , Feil Edward J. , Blanc Dominique S. 
Published: January 2016

ABSTRACT

Whole-genome sequencing (WGS) of 228 isolates was used to elucidate the origin and dynamics of a long-term outbreak of methicillin-resistant
Staphylococcus aureus
(MRSA) sequence type 228 (ST228) SCC
mec
I that involved 1,600 patients in a tertiary care hospital between 2008 and 2012. Combining of the sequence data with detailed metadata on patient admission and movement confirmed that the outbreak was due to the transmission of a single clonal variant of ST228, rather than repeated introductions of this clone into the hospital. We note that this clone is significantly more frequently recovered from groin and rectal swabs than other clones (
P
< 0.0001) and is also significantly more transmissible between roommates (
P
< 0.01). Unrecognized MRSA carriers, together with movements of patients within the hospital, also seem to have played a major role. These atypical colonization and transmission dynamics can help explain how the outbreak was maintained over the long term. This “stealthy” asymptomatic colonization of the gut, combined with heightened transmissibility (potentially reflecting a role for environmental reservoirs), means the dynamics of this outbreak share some properties with enteric pathogens such as vancomycin-resistant enterococci or
Clostridium difficile
.

IMPORTANCE
Using whole-genome sequencing, we showed that a large and prolonged outbreak of methicillin-resistant
Staphylococcus aureus
was due to the clonal spread of a specific strain with genetic elements adapted to the hospital environment. Unrecognized MRSA carriers, the movement of patients within the hospital, and the low detection with clinical specimens were also factors that played a role in this occurrence. The atypical colonization of the gut means the dynamics of this outbreak may share some properties with enteric pathogens.
Growth data of underprivileged children living in rural areas of Chin State, Burma/Myanmar, compared to the WHO reference growth standards: an observational study
Authors: Prenkert Malin , Ehnfors Margareta 
Published: January 2016

ObjectivesTo explore growth data (height-for-age, weight-for-age and BMI-for-age) of children living in poor socioeconomic conditions in rural areas of Chin State, Burma/Myanmar; and to compare these data with the growth and development z-score (GDZ) values for school-aged children and adolescents, provided by the WHO.SettingA support and educational programme, run by the Swedish association Chin Development and Research Society (CDRS), was carried out among underprivileged school-aged children, unable to attend school without economic and practical support, living in villages and remote areas in Chin State.ParticipantsCommunity leaders who were well familiar with the citizens in the community identified children in need of this support. Other community members could also suggest or apply for this. The sample includes all participating children in the CDRS programme at the time of the data collection in six townships. The children were placed in host families, close to a suitable school. Two samples with a total of 639 children from 144 villages and remote areas were obtained:1. Children in the CDRS Chin Programme (CCP) (2007–2010) comprised 558 children: 50% girls and boys.2. Children in the Chin Society (CCS) (2010) comprised 81 children: 44% girls and 56% boys.Primary outcome measuresGrowth data.ResultsAll growth data from both groups deviated significantly from the WHO standard references (p≤0.001). The prevalence of stunting (height-for-age ≤–2SD) was 52% among girls and 68% among boys. High levels of wasting (weight-for-age ≤–2SD) were found among girls 29% and boys 36% aged 5–10 years. In addition, severe thinness (BMI-for-age ≤–2SD) was found among girls 31% and boys 44%, all results to be compared to the expected 2.27%.ConclusionsMany more than expected—according to the WHO reference values—in CCP and CCS suffered from stunting, wasting and thinness.
Rigorous anaesthesia management protocol for patients with intracranial arterial stenosis: a prospective controlled-cohort study
Authors: Laiwalla Azim N , Ooi Yinn Cher , Van De Wiele Barbara , Ziv Keren , Brown Adam , Liou Raymond , Saver Jeffrey L , Gonzalez Nestor R 
Published: January 2016

ObjectivesReducing variability is integral in quality management. As part of the ongoing Encephaloduroarteriosynangiosis Revascularisation for Symptomatic Intracranial Arterial Stenosis (ERSIAS) trial, we developed a strict anaesthesia protocol to minimise fluctuations in patient parameters affecting cerebral perfusion. We hypothesise that this protocol reduces the intraoperative variability of targeted monitored parameters compared to standard management.DesignProspective cohort study of patients undergoing encephaloduroarteriosynangiosis surgery versus standard neurovascular interventions. Patients with ERSIAS had strict perioperative management that included normocapnia and intentional hypertension. Control patients received regular anaesthetic standard of care. Minute-by-minute intraoperative vitals were electronically collected. Heterogeneity of variance tests were used to compare variance across groups. Mixed-model regression analysis was performed to establish the effects of treatment group on the monitored parameters.SettingTertiary care centre.Participants24 participants: 12 cases (53.8 years±16.7 years; 10 females) and 12 controls (51.3 years±15.2 years; 10 females). Adults aged 30–80 years, with transient ischaemic attack or non-disabling stroke (modified Rankin Scale <3) attributed to 70–99% intracranial stenosis of the carotid or middle cerebral artery, were considered for enrolment. Controls were matched according to age, gender and history of neurovascular intervention.Main outcome measuresVariability of heart rate, mean arterial blood pressure (MAP), systolic blood pressure and end tidal CO2(ETCO2) throughout surgical duration.ResultsThere were significant reductions in the intraoperative MAP SD (4.26 vs 10.23 mm Hg; p=0.007) and ETCO2 SD (0.94 vs 1.26 mm Hg; p=0.05) between the ERSIAS and control groups. Median MAP and ETCO2 in the ERSIAS group were higher (98 mm Hg, IQR 23 vs 75 mm Hg, IQR 15; p<0.001, and 38 mm Hg, IQR 4 vs 32 mm Hg, IQR 3; p<0.001, respectively).ConclusionsThe ERSIAS anaesthesia protocol successfully reduced intraoperative fluctuations of MAP and ETCO2. The protocol also achieved normocarbia and the intended hypertension.Trial registration numberNCT01819597; Pre-results.
The efficacy of a movement control exercise programme to reduce injuries in youth rugby: a cluster randomised controlled trial
Authors: Hislop M D , Stokes K A , Williams S , McKay C D , England M , Kemp S P T , Trewartha G 
Published: January 2016

BackgroundInjuries to youth rugby players have become an increasingly prominent health concern, highlighting the importance of developing and implementing appropriate preventive strategies. A growing body of evidence from other youth sports has demonstrated the efficacy of targeted exercise regimens to reduce injury risk. However, studies have yet to investigate the effect of such interventions in youth contact sport populations like rugby union.ObjectiveTo determine the efficacy of an evidence-based movement control exercise programme compared with a sham exercise programme to reduce injury risk in youth rugby players. Exercise programme compliance between trial arms and the effect of coach attitudes on compliance will also be evaluated.SettingSchool rugby coaches in England will be the target of the researcher intervention, with the effects of the injury prevention programmes being measured in male youth players aged 14–18 years in school rugby programmes over the 2015–2016 school winter term.MethodsA cluster-randomised controlled trial with schools randomly allocated to either a movement control exercise programme or a sham exercise programme, both of which are coach-delivered. Injury measures will derive from field-based injury surveillance, with match and training exposure and compliance recorded. A questionnaire will be used to evaluate coach attitudes, knowledge, beliefs and behaviours both prior to and on the conclusion of the study period.Outcome measuresSummary injury measures (incidence, severity and burden) will be compared between trial arms, as will the influence of coach attitudes on compliance and injury burden. Additionally, changes in these outcomes through using the exercise programmes will be evaluated.Trial registration numberISRTCNN13422001.
Evaluating a DVD promoting breast cancer awareness among black women aged 25–50 years in East London
Authors: Greenhough Beth , Dembinsky Melanie , Dyck Isabel , Brown Tim , Robson John , Homer Kate , Sajani Cynthia , Carter Lucy , Duffy Stephen W , Ornstein Mark 
Published: January 2016

BackgroundThe mean age of presentation for breast cancer among black women is substantially earlier than their white counterparts. Black women also present with adverse prognostic factors that have major clinical implications, including lower survival. To pilot the use of a 6 min DVD on breast cancer in young (under 50 years) black women, to raise awareness and examine the impact of the DVD on increased consultation and referral rates among these women.MethodsTwo general practices (intervention practices) in the Hackney area were randomised to have the DVD mailed to all black women aged 25–50 years registered with the practices, and two practices to no intervention (control practices). EMIS data was used to compare consultation rates preintervention and postintervention, in the intervention as well as control practices. Interviews with practice staff and focus groups with patients in participating practices provided qualitative data on the study context and DVD effectiveness.ResultsA trend of declining consultations for breast symptoms was observed (−22% and −31% among non-black women in the control and intervention practices, and −23% among black women in the control practice) except among the target population of black women aged 25–50 years for the DVD in the intervention practices, which saw an increase of 28% in consultations. The qualitative data indicated that the DVD was well received in the target population, and suggested further ways of disseminating awareness messages and overcoming barriers to help-seeking.ConclusionsPilot results suggest that the strategy of distributing the DVD may increase consultations for breast problems.
Expert consensus on dynamics of laboratory tests for diagnosis of macrophage activation syndrome complicating systemic juvenile idiopathic arthritis
Authors: Ravelli Angelo , Minoia Francesca , Davì Sergio , Horne AnnaCarin , Bovis Francesca , Pistorio Angela , Aricò Maurizio , Avcin Tadej , Behrens Edward M , De Benedetti Fabrizio , Filipovic Alexandra , Grom Alexei A , Henter Jan-Inge , Ilowite Norman T , Jordan Michael B , Khubchandani Raju , Kitoh Toshiyuki , Lehmberg Kai , Lovell Daniel J , Miettunen Paivi , Nichols Kim E , Ozen Seza , Pachlopnik Schmid Jana , Ramanan Athimalaipet V , Russo Ricardo , Schneider Rayfel , Sterba Gary , Uziel Yosef , Wallace Carol , Wouters Carine , Wulffraat Nico , Demirkaya Erkan , Brunner Hermine I , Martini Alberto , Ruperto Nicolino , Cron Randy Q 
Published: January 2016

ObjectiveTo identify which laboratory tests that change over time are most valuable for the timely diagnosis of macrophage activation syndrome (MAS) complicating systemic juvenile idiopathic arthritis (sJIA).MethodsA multistep process, based on a combination of expert consensus and analysis of real patient data, was conducted. A panel of experts was first asked to evaluate 115 profiles of patients with MAS, which included the values of laboratory tests at the pre-MAS visit and at MAS onset, and the change in values between the two time points. The experts were asked to choose the 5 laboratory tests in which change was most important for the diagnosis of MAS and to rank the 5 selected tests in order of importance. The relevance of change in laboratory parameters was further discussed and ranked by the same experts at a consensus conference.ResultsPlatelet count was the most frequently selected test, followed by ferritin level, aspartate aminotransferase (AST), white cell count, neutrophil count, and fibrinogen and erythrocyte sedimentation rate. Ferritin was most frequently assigned the highest score. At the end of the process, platelet count, ferritin level and AST were the laboratory tests in which the experts found change over time to be most important.ConclusionsWe identified the laboratory tests in which change over time is most valuable for the early diagnosis of MAS in sJIA. The dynamics of laboratory values during the course of MAS should be further scrutinised in a prospective study in order to establish the optimal cut-off values for their variation.
Mastermind-Like 3 Controls Proliferation and Differentiation in Neuroblastoma
Authors: Heynen Guus J.J.E. , Nevedomskaya Ekaterina , Palit Sander , Jagalur Basheer Noorjahan , Lieftink Cor , Schlicker Andreas , Zwart Wilbert , Bernards Rene , Bajpe Prashanth Kumar 
Published: January 2016

Abstract
Neuroblastoma cell lines can differentiate upon treatment with retinoic acid (RA), a finding that provided the basis for the clinical use of RA to treat neuroblastoma. However, resistance to RA is often observed, which limits its clinical utility. Using a gain-of-function genetic screen, we identified an unexpected link between RA signaling and mastermind-like 3 (MAML3), a known transcriptional coactivator for NOTCH. Our findings indicate that MAML3 expression leads to the loss of activation of a subset of RA target genes, which hampers RA-induced differentiation and promotes resistance to RA. The regulatory DNA elements of this subset of RA target genes show overlap in binding of MAML3 and the RA receptor, suggesting a direct role for MAML3 in the regulation of these genes. In addition, MAML3 has RA-independent functions, including the activation of IGF1R and downstream AKT signaling via upregulation of IGF2, resulting in increased proliferation. These results demonstrate an important mechanistic role for MAML3 in proliferation and RA-mediated differentiation.
Implications: MAML3 coordinates transcription regulation with receptor tyrosine kinase pathway activation, shedding new light on why this gene is mutated in multiple cancers. Mol Cancer Res; 14(5); 411–22. ©2016 AACR.
Regulatory T Cells from Colon Cancer Patients Inhibit Effector T-cell Migration through an Adenosine-Dependent Mechanism
Authors: Sundström Patrik , Stenstad Hanna , Langenes Veronica , Ahlmanner Filip , Theander Lisa , Ndah Tapuka Gordon , Fredin Kamilla , Börjesson Lars , Gustavsson Bengt , Bastid Jérémy , Quiding-Järbrink Marianne 
Published: January 2016

AbstractT cell–mediated immunity is a major component of antitumor immunity. In order to be efficient, effector T cells must leave the circulation and enter into the tumor tissue. Regulatory T cells (Treg) from gastric cancer patients, but not from healthy volunteers, potently inhibit migration of conventional T cells through activated endothelium. In this study, we compared T cells from colon cancer patients and healthy donors to determine the mechanisms used by Tregs from cancer patients to inhibit conventional T-cell migration. Our results showed that circulating Tregs from cancer patients expressed high levels of CD39, an ectoenzyme mediating hydrolysis of ATP to AMP, as a rate-determining first step in the generation of immunosuppressive adenosine. Tumor-associated Tregs expressed even more CD39, and we therefore examined the importance of adenosine in Treg-mediated inhibition of T-cell transendothelial migration in vitro. Exogenous adenosine significantly reduced migration of conventional T cells from healthy volunteers, and blocking either adenosine receptors or CD39 enzymatic activity during transmigration restored the ability of conventional T cells from cancer patients to migrate. Adenosine did not directly affect T cells or endothelial cells, but reduced the ability of monocytes to activate the endothelium. Taken together, our results indicate that Treg-derived adenosine acts on monocytes and contributes to reduced transendothelial migration of effector T cells into tumors. This effect of Tregs is specific for cancer patients, and our results indicate that Tregs may affect not only T-cell effector functions but also their migration into tumors. Cancer Immunol Res; 4(3); 183–93. ©2016 AACR.