bluuebunny/crossref_metadata_2025 huggingface dataset:
Midtrimester spontaneous torsion of unruptured gravid rudimentary horn: Presurgical diagnosis on magnetic resonance imaging
Authors: Singh Pragya , Gupta Rohini , Das Banashree , Bajaj Sunil Kumar , Misra Ritu
Published: May 2015

Unicornuate uterus with rudimentary horn occurs due to failure of complete development and partial fusion of one of the Müllerian ducts. Pregnancy in a non‐communicating rudimentary horn is extremely rare, with a reported incidence of 1 in 76 000–150 000 pregnancies, and usually terminates in rupture during the first or second trimester. Clinical diagnosis of rudimentary horn pregnancy in a woman with history of normal vaginal delivery in prior gestations is difficult. The role of sonography, and more recently, magnetic resonance imaging, in the presurgical diagnosis of rudimentary horn pregnancy is established. We present a case of magnetic resonance imaging diagnosis of 20‐week pregnancy in the unruptured non‐communicating rudimentary horn in a patient with previous history of two full‐term normal vaginal deliveries. The novelty of the case lies in the fact that there was associated torsion of the gravid rudimentary horn and ipsilateral ovary, which has not been reported previously.
Accuracy of visual inspection with acetic acid and with Lugol's iodine for cervical cancer screening: Meta‐analysis
Authors: Qiao Liang , Li Bo , Long Mei , Wang Xiao , Wang Anrong , Zhang Guonan
Published: May 2015

The aim of this review was to provide an updated summary estimation of the accuracy of visual inspection with acetic acid (VIA) and with Lugol's iodine (VILI) in detecting cervical cancer and precancer. Studies on VIA/VILI accuracy were eligible in which VIA/VILI was performed on asymptomatic women who all underwent confirmatory testing of histology, combination of colposcopy and histology, or combination of multiple screening tests, colposcopy and histology, to detect cervical intraepithelial neoplasia grade 2 or worse (CIN2+ or CIN3+). A bivariate model was fitted to estimate the accuracy of VIA/VILI and provide estimates of heterogeneity. Subgroup analysis was used to investigate the source of heterogeneity. A total of 29 studies on VIA and 19 studies on VILI were included finally in the meta‐analysis. The summary sensitivity and specificity of VIA for CIN2+ were 73.2% (95%CI: 66.5–80.0%) and 86.7% (95%CI: 82.9–90.4%), respectively, and those for VILI were 88.1% (95%CI: 81.5–94.7%) and 85.9% (95%CI: 81.7–90.0%), respectively. VIA and VILI were both more sensitive in detecting more severe outcome, although there was a slight loss in specificity. Apparent heterogeneity existed in sensitivity and specificity for both VIA and VILI. High sensitivity of both VIA and VILI for CIN2+ was found when a combination of colposcopy and histology was used as disease confirmation. VIA, VILI, even a combination of them in parallel, could be good options for cervical screening in low‐resource settings. Significant differences in sensitivity between different gold standards might provide a proxy for optimization of ongoing cervical cancer screening programs.
Improving Access to Psychological Therapies (IAPT) and treatment outcomes: Epistemological Assumptions and Controversies
Authors: Williams C. H. J.
Published: May 2015

Accessible summary

Cognitive behaviour therapy (CBT) is recommended as a primary treatment choice in England, for anxiety and depression, by the National Institute for Health and Care Excellence (NICE).

It has been argued that CBT has enjoyed political and cultural dominance and this has arguably led to maintained government investment in England for the cognitive and behavioural treatment of mental health problems. The government programme ‘Improving Access to Psychological Therapies’ (IAPT) aims to improve the availability of CBT.

The criticism of the NICE evidence‐based guidelines supporting the IAPT programme, has been the dominance of the gold standard randomized controlled trial methodology, with a focus on numerical outcome data, rather than a focus on a recovery narrative. RCT‐based research is influenced by a philosophical paradigm called positivism.

The IAPT culture is arguably influenced by one research paradigm and such an influence can skew services only towards numerical outcome data as the only truth of ‘recovery’. An interpretative paradigm could assist in shaping service‐based cultures, alter how services are evaluated and improve the richness of CBT research.

This paper explores the theory of knowledge (epistemology) that underpins the evidence‐based perspective of CBT and how this influences service delivery. The paper argues that the inclusion of service user narrative (qualitative data) can assist the evaluation of CBT from the user's perspective and can understand the context in which people live and how they access services. A qualitative perspective is discussed as a research strategy, capturing the lived experience of under‐represented groups, such as sexual, gender and ethnic minorities.

Cognitive behaviour therapy (CBT) has enjoyed political and cultural dominance within mental healthcare, with renewed government investment in England for the‘Improving Access to Psychological Therapies’ (IAPT) programme. The criticism of the evidence‐based guidelines, published by the National Institute for Health and Care Excellence (NICE), which supports the IAPT programme has been the dominance of the gold standard randomized controlled trial methodology. The definition of ‘recovery’ used by IAPT is based on a positivist position, with a focus on numerical outcome data garnered through psychometric measures. An interpretative perspective of recovery, which would include a subjective individual patient/service user narrative and would include a collaborative qualitative dialogue, is arguably absent from the IAPT programme. The challenge inherent in the IAPT programme is the high demand/high turnover culture, and psychometric measures are quick to administer; however, this culture is driven from one research paradigm. An interpretative paradigm may assist in shaping service‐based cultures, alter how services are evaluated, and improve the richness of CBT research.
Long‐term presence and habitat use of Cuvier's beaked whale (Ziphius cavirostris) in the Central Tyrrhenian Sea
Authors: Arcangeli Antonella , Campana Ilaria , Marini Luca , MacLeod Colin D.
Published: May 2015

Cuvier's beaked whale (Ziphius cavirostris, G. Cuvier 1823) is a poorly known species and many international agreements have asked for a better understanding of its biology for conservation purposes. In the present study, systematic cetacean surveys were carried out from ferries along a trans‐regional fixed transect in the Central Tyrrhenian Sea (Civitavecchia, Latium – Golfo Aranci, Sardinia), just outside the southeastern border of the Pelagos Sanctuary. This research provided long‐term, consistent data on Cuvier's beaked whale during two research periods (1990–1992 and 2007–2011). The objective of the research was to compare the presence, distribution and habitat use of Cuvier's beaked whale between the two investigated periods. Summer data (June–September) from the two periods were compared in terms of frequency of sightings, group size and spatial distribution related to the main ecogeographical features. A presence‐absence model (generalized additive modelling) was performed to predict habitat suitability in the two study periods. The results highlight long‐term site fidelity of Cuvier's beaked whale in the Central Tyrrhenian Sea with encounter rates comparable to the ones reported for other key areas. Separate suitability models based on 1990s and 2000s data appeared to work for each individual time period but differences were evident between the two periods, indicating changes in habitat selection over time. Our findings of the study appear to expand the definition of suitable beaked whale habitat and underline how the temporal scale of the analysis can affect the results in habitat studies. Moreover, this research highlights the importance of the Central Tyrrhenian Sea marine region for Cuvier's beaked whale and the ability of continuous monitoring to identify changes in cetacean frequency and distribution, necessary for adaptive conservation management approaches.
Population genomic analyses reveal possible drivers of population divergence
Authors: Skrede Inger , Brandström Durling Mikael
Published: May 2015

Recent advances in sequencing technology and efficiency enable new and improved methods to investigate how populations diverge and species evolve. Fungi have relatively small and simple genomes and can often be cultured in the laboratory. Fungal populations can thus be sequenced for a relatively low cost, which makes them ideal for population genomic analyses. In several recent population genomic studies, wild populations of fungal model organisms and human pathogens have been analysed, for example Neurospora crassa (Ellison et al. ), Saccharomyces uvarum (Almeida et al. ), Coccidioides spp. (Neafsey et al. ) and Cryptococcus gatti (Engelthaler et al. ). In this issue of Molecular Ecology, Branco et al. () apply population genomic tools to understand population divergence and adaptation in a symbiotic (mycorrhizal) fungus. This study exemplifies the possibilities of diving deeper into the genomic features involved in population divergence and speciation, also for nonmodel organisms, and how molecular and analytical tools will improve our understanding of the patterns and mechanisms that underlie adaptation to habitats, population divergence and dispersal limitation of fungi.
Fritillaria avromanica sp. nov. (Liliaeceae) from Iran and notes on F. melananthera in Turkey
Authors: Advay Mahfouz , Tekşen Mehtap , Maroofi Hosein
Published: May 2015

Fritillaria avromanica is described as a new species from Iran based on specimens collected from Hawraman. Taxonomic descriptions, illustrations, geographical distribution, habitat, ecology, and conservation status of the new species and the closely related F. assyriaca and F. melananthera are provided. In addition F. melananthera is given the rank of species, rather than being subspecies of F. assyriaca. The differences between F. melananthera and F. assyriaca are discussed with pollen and chromosomal characteristics.
Morphometry of the hard palate in Down's syndrome through CBCT‐image analysis
Authors: Abeleira M. T. , Outumuro M. , Diniz M. , Limeres J. , Ramos I. , Diz P.
Published: May 2015

Structured Abstract

Objectives

To define the morphometry of the hard palate in Down's syndrome (DS) on cone beam computed tomography (CBCT) images.

Setting and Sample Population

Santiago de Compostela University (Spain). The study group included 40 white DS individuals aged 10 to 40 years (mean = 18.8 ± 7.3 years), 25 males and 15 females. The control group consisted of 40 individuals matched for age and sex were selected.

Material & Methods

Nine measurements were taken on the CBCT images. Axial plane: anteroposterior length (aAPL) and arch length (aARL); sagittal plane: anteroposterior length (sAPL), maximum height (sMH) and sagittal arch (sAR); coronal plane: interdental width (cIDW), height (cHE), skeletal width (cSW) and coronal arch (cAR).

Results

aAPL, aARL, sAPL, sMH, sAR, cMH and cAR were comparable in the two groups. cIDW and cSW were greater in controls than in DS. We found no statistically significant differences between males and females with DS. In the controls, sAPL and sAR were greater in males than females. In DS, age only had a statistically significantly increasing effect on aAPL and sAPL. In the controls, age significantly affected sAR and cHE.

Conclusion

The hard palate is narrower in DS than in controls, but the anteroposterior measurements and the height of the vault are comparable in both groups.
New prenyllipid metabolites identified in Arabidopsis during photo‐oxidative stress
Authors: Dłużewska Jolanta , Zieliński Kamil , Nowicka Beatrycze , Szymańska Renata , Kruk Jerzy
Published: May 2015

In the present study, we have identified new prenyllipid metabolites formed during high light stress in Arabidopsis thaliana, whose origin and function remained unknown so far. It was found that plastoquinone‐C accumulates mainly in the reduced form under high light conditions, as well as during short‐term excess light illumination both in the wild‐type and tocopherol biosynthetic vte1 mutant, suggesting that plastoquinone‐C, a singlet oxygen‐derived prenyllipid, is reduced in chloroplasts by photosystem II or enzymatically, outside thylakoids. Plastoquinone‐B, a fatty acid ester of plastoquinone‐C, was identified for the first time in Arabidopsis in high light grown wild‐type plants and during short‐time, excess light illumination of the wild‐type plants and the vte1 mutant. The gene expression analysis showed that vte2 gene is most pronouncedly up‐regulated among the prenyllipid biosynthetic genes under high light and induction of its expression is mainly caused by an increased level of singlet oxygen, as was demonstrated in experiments with D2O‐treated plants under excess light conditions.
Magnetic resonance tagging for diagnosis of focal hypertrophic cardiomyopathy in a child
Authors: Çoban Gökçen , Yıldırım Müge Afşar , Dönmez Fuldem Yıldırım , Tarhan Nefise Çağla , Erdoğan İlkay , Coşkun Mehmet
Published: May 2015

Magnetic resonance imaging has become an important diagnostic tool in the differential diagnosis of lesions for evaluation of cardiovascular disorders. In magnetic resonance tagging (MRt), tissue elements are magnetically labeled so that their positions can be tracked as a function of time. Thus, MRt evaluates heart wall motion both qualitatively and quantitatively. We present herein the case of a 12‐year‐old boy who had chest pain, dyspnea on effort and murmur. On cardiac computed tomography, there was focal thickening of the left ventricular posterior wall, similar to a mass. MRt indicated active displacement and deformation of the tags at the level of the hypertrophic myocardium during systole, as with normal myocardium. Thus, the tagged images supported the diagnosis of focal hypertrophic cardiomyopathy (HCM). In view of these results, MRt should be considered as a useful technique for differentiating between a mass‐like focal lesion such as neoplasm and HCM.
Two siblings with cortical dysplasia: Clinico‐electroencephalographic features
Authors: Fukasawa Tatsuya , Kubota Tetsuo , Negoro Tamiko , Maruyama Shinsuke , Honda Ryoko , Saito Yuko , Itoh Masayuki , Kakita Akiyoshi , Sugai Kenji , Otsuki Taisuke , Kato Mitsuhiro , Natsume Jun , Watanabe Kazuyoshi
Published: May 2015

The older of two siblings began to have spasms and partial seizures at 1 month of age. Head magnetic resonance imaging showed an abnormal area in the left temporo‐parieto‐occipital region. Interictal electroencephalogram (EEG) showed a suppression–burst pattern. Adrenocorticotropic hormone stopped the spasms, but the seizures continued. Clonazepam, carbamazepine, zonisamide, and clobazam were ineffective. She underwent focal resection at age 8 months. Postoperatively, the seizures disappeared. Histopathologically, the lesion appeared to be focal cortical dysplasia typeIIa. The younger sibling had spasms from birth. Head magnetic resonance imaging showed left hemi‐megalencephaly. InterictalEEGshowed a suppression‐burst pattern. Phenobarbital, valproic acid, and zonisamide were ineffective. He underwent hemispherotomy at age 2 months and became seizure free. The histopathological features were consistent with those of hemi‐megalencephaly. The siblings’EEGand clinical courses had some similarities. These siblings’ conditions may have the same genetic background.
How Narrow is Aristotle's Contemplative Ideal?
Authors: Walker Matthew D.
Published: May 2015

InNicomachean EthicsX.7–8, Aristotle defends a striking view about the good for human beings. According to Aristotle, the single happiest way of life is organized around philosophical contemplation. According tothe narrowness worry, however, Aristotle's contemplative ideal is unduly Procrustean, restrictive, inflexible, and oblivious of human diversity. In this paper, I argue that Aristotle has resources for responding to the narrowness worry, and that his contemplative ideal can take due account of human diversity.
Evaluation of the effectiveness of a pathogen inactivation technology against clinically relevant transfusion‐transmitted bacterial strains
Authors: Schmidt Michael , Hourfar Michael K. , Sireis Walid , Pfeiffer Ulrich , Göttig Stephan , Kempf Volkhard A.J. , McDonald Carl P. , Seifried Erhard
Published: May 2015

BACKGROUND

To increase blood safety, various procedures are currently implemented, including donor selection, optimized donor arm disinfection, and diversion. In addition, pathogen inactivation (PI) techniques can be used for platelets (PLTs) and plasma concentrates.

STUDY DESIGN AND METHODS

This study investigated the clinical efficacy of an inactivation technique for different blood components at two time points (12 and 35.5 hr). Eight transfusion‐relevant bacterial strains were spiked at two different concentrations (100 and 1000 colony‐forming units [CFUs]/bag) into whole blood (WB), apheresis PLTs (APs), and buffy coat (BC)‐derived minipool PLTs.

RESULTS

The bacterial concentrations were higher than 106 CFUs/mL within 24 hours after spiking depending on the particular bacterial strain. PI was absolute for all of the APs performed 12 hours after inoculation, but the bacterial strains of Klebsiella pneumoniae and Bacillus cereus were not completely inactivated in WB or BC PLTs, performed 35.5 and 12 hours after inoculation, respectively.

CONCLUSION

The INTERCEPT PI system was not 100% effective for high concentrations of certain K. pneumoniae strains or spore‐forming B. cereus. A critical observation was that the period between blood donation and inactivation needs to be minimal to enable efficient PI. In the case where PI cannot be performed immediately after preparation, a combination of a PI technology after the production of blood components with a rapid bacterial screen test on Day 4 or 5 after donation may offer a solution to further prevent the risk of bacterial transmission by transfusion.
The water‐energy nexus: future water resource availability and its implications on UK thermal power generation
Authors: Murrant Daniel , Quinn Andrew , Chapman Lee
Published: May 2015

An increasing population coupled with the uncertain, but increasingly likely, impacts of climate change have led to a heightened level of global academic attention to the interdependencies that exist between the water and energy infrastructure networks. However, to date there has been limited research considering the water‐energy nexus within a UK context. This article reviews the global and national literature to identify how a future lack of available water resource will impact upon the UK thermal power generation fleet, both in terms of freshwater resource and environmental constraints. It concludes that a combination of freshwater resource management and adaptation to use alternative water sources will be key in mitigating and adapting to climate impacts.
Unprecedented Silver Resistance in Clinically Isolated Enterobacteriaceae: Major Implications for Burn and Wound Management
Authors: Finley Phillip J. , Norton Rhy , Austin Cindy , Mitchell Amber , Zank Sara , Durham Paul
Published: May 2015

Increased utilization of inorganic silver as an adjunctive to many medical devices has raised concerns of emergent silver resistance in clinical bacteria. Although the molecular basis for silver resistance has been previously characterized, to date, significant phenotypic expression of these genes in clinical settings is yet to be observed. Here, we identified the first strains of clinical bacteria expressing silver resistance at a level that could significantly impact wound care and the use of silver-based dressings. Screening of 859 clinical isolates confirmed 31 harbored at least 1 silver resistance gene. Despite the presence of these genes, MIC testing revealed most of the bacteria displayed little or no increase in resistance to ionic silver (200 to 300 μM Ag
+
). However, 2 isolates (
Klebsiella pneumonia
and
Enterobacter cloacae
) were capable of robust growth at exceedingly high silver concentrations, with MIC values reaching 5,500 μM Ag
+
. DNA sequencing of these two strains revealed the presence of genes homologous to known genetic determinants of heavy metal resistance. Darkening of the bacteria's pigment was observed after exposure to high silver concentrations. Scanning electron microscopy images showed the presence of silver nanoparticles embedded in the extracellular polymeric substance of both isolates. This finding suggested that the isolates may neutralize ionic silver via reduction to elemental silver. Antimicrobial testing revealed both organisms to be completely resistant to many commercially available silver-impregnated burn and wound dressings. Taken together, these findings provide the first evidence of clinical bacteria capable of expressing silver resistance at levels that could significantly impact wound management.
In Vitro
and
In Vivo
Efficacy of Amphotericin B Combined with Posaconazole against Experimental Disseminated Sporotrichosis
Authors: Mario Débora Nunes , Guarro Josep , Santurio Janio Morais , Alves Sydney Hartz , Capilla Javier
Published: May 2015

We evaluated the combination of posaconazole with amphotericin B
in vitro
and in a murine model of systemic infections caused by
Sporothrix brasiliensis
and
Sporothrix schenckii
sensu stricto
.
In vitro
data demonstrated a synergistic effect, and although posaconazole alone was effective against sporotrichosis, efficacy in terms of survival and burden reduction was increased with the combination. This combination might be an option against disseminated sporotrichosis, especially when itraconazole or amphotericin B at optimal doses are contraindicated.
Effect of Lactobacillus rhamnosus GG Administration on Vancomycin-Resistant Enterococcus Colonization in Adults with Comorbidities
Authors: Doron Shira , Hibberd Patricia L. , Goldin Barry , Thorpe Cheleste , McDermott Laura , Snydman David R.
Published: May 2015

Vancomycin-resistant enterococci (VRE) are endemic in health care settings. These organisms colonize the gastrointestinal tract and can lead to infection which is associated with increased mortality. There is no treatment for VRE colonization. We conducted a randomized, double-blind, placebo-controlled clinical trial to examine the safety and efficacy of administration of the probiotic
Lactobacillus rhamnosus
GG (LGG) for the reduction or elimination of intestinal colonization by VRE. Colonized adults were randomized to receive LGG or placebo for 14 days. Quantitative stool cultures for LGG and VRE were collected at baseline and days 7, 14, 21, 28, and 56. Day 14 stool samples from some subjects were analyzed by quantitative PCR (qPCR) for LGG. Patients were closely monitored for adverse events. Eleven subjects, of whom 5 received LGG and 6 received placebo, were analyzed. No differences in VRE colony counts were seen at any time points between groups. No decline in colony counts was seen over time in subjects who received LGG. LGG was detected by PCR in all samples tested from subjects who received LGG but was only isolated in culture from 2 of 5 subjects in the LGG group. No treatment-related adverse events were seen. We demonstrated that LGG could be administered safely to patients with comorbidities and is recoverable in some patients' stool cultures. Concomitant administration of antibiotics may have resulted in an inability to recover viable organisms from stool samples, but LGG DNA could still be detected by qPCR. LGG administration did not affect VRE colonization in this study. (This study was registered at Clinicaltrials.gov under registration no. NCT00756262.)
Ex Vivo
Drug Susceptibility Testing and Molecular Profiling of Clinical Plasmodium falciparum Isolates from Cambodia from 2008 to 2013 Suggest Emerging Piperaquine Resistance
Authors: Chaorattanakawee Suwanna , Saunders David L. , Sea Darapiseth , Chanarat Nitima , Yingyuen Kritsanai , Sundrakes Siratchana , Saingam Piyaporn , Buathong Nillawan , Sriwichai Sabaithip , Chann Soklyda , Se Youry , Yom You , Heng Thay Kheng , Kong Nareth , Kuntawunginn Worachet , Tangthongchaiwiriya Kuntida , Jacob Christopher , Takala-Harrison Shannon , Plowe Christopher , Lin Jessica T. , Chuor Char Meng , Prom Satharath , Tyner Stuart D. , Gosi Panita , Teja-Isavadharm Paktiya , Lon Chanthap , Lanteri Charlotte A.
Published: May 2015

Cambodia's first-line artemisinin combination therapy, dihydroartemisinin-piperaquine (DHA-PPQ), is no longer sufficiently curative against multidrug-resistant
Plasmodium falciparum
malaria at some Thai-Cambodian border regions. We report recent (2008 to 2013) drug resistance trends in 753 isolates from northern, western, and southern Cambodia by surveying for
ex vivo
drug susceptibility and molecular drug resistance markers to guide the selection of an effective alternative to DHA-PPQ. Over the last 3 study years, PPQ susceptibility declined dramatically (geomean 50% inhibitory concentration [IC
50
] increased from 12.8 to 29.6 nM), while mefloquine (MQ) sensitivity doubled (67.1 to 26 nM) in northern Cambodia. These changes in drug susceptibility were significantly associated with a decreased prevalence of
P. falciparum
multidrug resistance 1 gene (Pf
mdr1
) multiple copy isolates and coincided with the timing of replacing artesunate-mefloquine (AS-MQ) with DHA-PPQ as the first-line therapy. Widespread chloroquine resistance was suggested by all isolates being of the
P. falciparum
chloroquine resistance transporter gene CVIET haplotype. Nearly all isolates collected from the most recent years had
P. falciparum
kelch13
mutations, indicative of artemisinin resistance.
Ex vivo
bioassay measurements of antimalarial activity in plasma indicated 20% of patients recently took antimalarials, and their plasma had activity (median of 49.8 nM DHA equivalents) suggestive of substantial
in vivo
drug pressure. Overall, our findings suggest DHA-PPQ failures are associated with emerging PPQ resistance in a background of artemisinin resistance. The observed connection between drug policy changes and significant reduction in PPQ susceptibility with mitigation of MQ resistance supports reintroduction of AS-MQ, in conjunction with monitoring of the
P. falciparum
mdr1
copy number, as a stop-gap measure in areas of DHA-PPQ failure.
Antibiofilm Activity of Low-Amperage Continuous and Intermittent Direct Electrical Current
Authors: Schmidt-Malan Suzannah M. , Karau Melissa J. , Cede Julia , Greenwood-Quaintance Kerryl E. , Brinkman Cassandra L. , Mandrekar Jayawant N. , Patel Robin
Published: May 2015

Bacterial biofilms are difficult to treat using available antimicrobial agents, so new antibiofilm strategies are needed. We previously showed that 20, 200, and 2,000 μA of electrical current reduced bacterial biofilms of
Staphylococcus aureus
,
Staphylococcus epidermidis
, and
Pseudomonas aeruginosa
. Here, we tested continuous direct current at lower amperages, intermittent direct current, and combinations of surface materials (Teflon or titanium) and electrode compositions (stainless steel, graphite, titanium, or platinum) against
S. aureus
,
S. epidermidis
, and
P. aeruginosa
biofilms. In addition, we tested 200 or 2,000 μA for 1 and 4 days against biofilms of 33 strains representing 13 species of microorganisms. The logarithmic reduction factor was used to measure treatment effects. Using continuous current delivery, the lowest active amperage was 2 μA for 1, 4, or 7 days against
P. aeruginosa
and 5 μA for 7 days against
S. epidermidis
and
S. aureus
biofilms. Delivery of 200 μA for 4 h a day over 4 days reduced
P. aeruginosa
,
S. aureus
, and
S. epidermidis
biofilms on Teflon or titanium discs. A reduction of
P. aeruginosa
,
S. aureus
, and
S. epidermidis
biofilms was measured for 23 of 24 combinations of surface materials and electrode compositions tested. Four days of direct current delivery reduced biofilms of 25 of 33 strains studied. In conclusion, low-amperage current or 4 h a day of intermittent current delivered using a variety of electrode compositions reduced
P. aeruginosa
,
S. aureus
, and
S. epidermidis
biofilms on a variety of surface materials. The electricidal effect was observed against a majority of bacterial species studied.
Frequency of Spontaneous Resistance to Peptide Deformylase Inhibitor GSK1322322 in Haemophilus influenzae, Staphylococcus aureus, Streptococcus pyogenes, and Streptococcus pneumoniae
Authors: Min Sharon , Ingraham Karen , Huang Jianzhong , McCloskey Lynn , Rilling Sarah , Windau Anne , Pizzollo Jason , Butler Deborah , Aubart Kelly , Miller Linda A. , Zalacain Magdalena , Holmes David J. , O'Dwyer Karen
Published: May 2015

The continuous emergence of multidrug-resistant pathogenic bacteria is compromising the successful treatment of serious microbial infections. GSK1322322, a novel peptide deformylase (PDF) inhibitor, shows good
in vitro
antibacterial activity and has demonstrated safety and efficacy in human proof-of-concept clinical studies.
In vitro
studies were performed to determine the frequency of resistance (FoR) to this antimicrobial agent in major pathogens that cause respiratory tract and skin infections. Resistance to GSK1322322 occurred at high frequency through loss-of-function mutations in the formyl-methionyl transferase (FMT) protein in
Staphylococcus aureus
(4/4 strains) and
Streptococcus pyogenes
(4/4 strains) and via missense mutations in
Streptococcus pneumoniae
(6/21 strains), but the mutations were associated with severe
in vitro
and/or
in vivo
fitness costs. The overall FoR to GSK1322322 was very low in
Haemophilus influenzae
, with only one PDF mutant being identified in one of four strains. No target-based mutants were identified from
S. pyogenes
, and only one or no PDF mutants were isolated in three of the four
S. aureus
strains studied. In
S. pneumoniae
, PDF mutants were isolated from only six of 21 strains tested; an additional 10 strains did not yield colonies on GSK1322322-containing plates. Most of the PDF mutants characterized from those three organisms (35/37 mutants) carried mutations in residues at or in close proximity to one of three highly conserved motifs that are part of the active site of the PDF protein, with 30 of the 35 mutations occurring at position V71 (using the
S. pneumoniae
numbering system).
Effects of a Mutation in the
gyrA
Gene on the Virulence of Uropathogenic Escherichia coli
Authors: Sánchez-Céspedes Javier , Sáez-López Emma , Frimodt-Møller N. , Vila Jordi , Soto Sara M.
Published: May 2015

Fluoroquinolones are among the drugs most extensively used for the treatment of bacterial infections in human and veterinary medicine. Resistance to quinolones can be chromosome or plasmid mediated. The chromosomal mechanism of resistance is associated with mutations in the DNA gyrase- and topoisomerase IV-encoding genes and mutations in regulatory genes affecting different efflux systems, among others. We studied the role of the acquisition of a mutation in the
gyrA
gene in the virulence and protein expression of uropathogenic
Escherichia coli
(UPEC). The HC14366M strain carrying a mutation in the
gyrA
gene (S83L) was found to lose the capacity to cause cystitis and pyelonephritis mainly due to a decrease in the expression of the
fimA
,
papA
,
papB
, and
ompA
genes. The levels of expression of the
fimA
,
papB
, and
ompA
genes were recovered on complementing the strain with a plasmid containing the
gyrA
wild-type gene. However, only a slight recovery was observed in the colonization of the bladder in the GyrA complement strain compared to the mutant strain in a murine model of ascending urinary tract infection. In conclusion, a mutation in the
gyrA
gene of uropathogenic
E. coli
reduced the virulence of the bacteria, likely in association with the effect of DNA supercoiling on the expression of several virulence factors and proteins, thereby decreasing their capacity to cause cystitis and pyelonephritis.
Evidence of Plasmodium falciparum Malaria Multidrug Resistance to Artemisinin and Piperaquine in Western Cambodia: Dihydroartemisinin-Piperaquine Open-Label Multicenter Clinical Assessment
Authors: Leang Rithea , Taylor Walter R. J. , Bouth Denis Mey , Song Lijiang , Tarning Joel , Char Meng Chuor , Kim Saorin , Witkowski Benoit , Duru Valentine , Domergue Anais , Khim Nimol , Ringwald Pascal , Menard Didier
Published: May 2015

Western Cambodia is recognized as the epicenter of
Plasmodium falciparum
multidrug resistance. Recent reports of the efficacy of dihydroartemisinin (DHA)-piperaquine (PP), the latest of the artemisinin-based combination therapies (ACTs) recommended by the WHO, have prompted further investigations. The clinical efficacy of dihydroartemisinin-piperaquine in uncomplicated falciparum malaria was assessed in western and eastern Cambodia over 42 days. Day 7 plasma piperaquine concentrations were measured and day 0 isolates tested for
in vitro
susceptibilities to piperaquine and mefloquine, polymorphisms in the
K13
gene, and the copy number of the
Pfmdr-1
gene. A total of 425 patients were recruited in 2011 to 2013. The proportion of patients with recrudescent infections was significantly higher in western (15.4%) than in eastern (2.5%) Cambodia (
P
<10
−3
). Day 7 plasma PP concentrations and median 50% inhibitory concentrations (IC
50
) of PP were independent of treatment outcomes, in contrast to median mefloquine IC
50
, which were found to be lower for isolates from patients with recrudescent infections (18.7 versus 39.7 nM;
P
= 0.005). The most significant risk factor associated with DHA-PP treatment failure was infection by parasites carrying the
K13
mutant allele (odds ratio [OR], 17.5; 95% confidence interval [CI], 1 to 308;
P
= 0.04). Our data show evidence of
P. falciparum
resistance to PP in western Cambodia, an area of widespread artemisinin resistance. New therapeutic strategies, such as the use of triple ACTs, are urgently needed and must be tested. (This study has been registered at the Australian New Zealand Clinical Trials Registry under registration no. ACTRN12614000344695.)
Borrelia burgdorferi, the Causative Agent of Lyme Disease, Forms Drug-Tolerant Persister Cells
Authors: Sharma Bijaya , Brown Autumn V. , Matluck Nicole E. , Hu Linden T. , Lewis Kim
Published: May 2015

Borrelia burgdorferi
is the causative agent of Lyme disease, which affects an estimated 300,000 people annually in the United States. When treated early, the disease usually resolves, but when left untreated, it can result in symptoms such as arthritis and encephalopathy. Treatment of the late-stage disease may require multiple courses of antibiotic therapy. Given that antibiotic resistance has not been observed for
B. burgdorferi
, the reason for the recalcitrance of late-stage disease to antibiotics is unclear. In other chronic infections, the presence of drug-tolerant persisters has been linked to recalcitrance of the disease. In this study, we examined the ability of
B. burgdorferi
to form persisters. Killing growing cultures of
B. burgdorferi
with antibiotics used to treat the disease was distinctly biphasic, with a small subpopulation of surviving cells. Upon regrowth, these cells formed a new subpopulation of antibiotic-tolerant cells, indicating that these are persisters rather than resistant mutants. The level of persisters increased sharply as the culture transitioned from the exponential to stationary phase. Combinations of antibiotics did not improve killing. Daptomycin, a membrane-active bactericidal antibiotic, killed stationary-phase cells but not persisters. Mitomycin C, an anticancer agent that forms adducts with DNA, killed persisters and eradicated growing and stationary cultures of
B. burgdorferi
. Finally, we examined the ability of pulse dosing an antibiotic to eliminate persisters. After addition of ceftriaxone, the antibiotic was washed away, surviving persisters were allowed to resuscitate, and the antibiotic was added again. Four pulse doses of ceftriaxone killed persisters, eradicating all live bacteria in the culture.
Artemether Exhibits Amoebicidal Activity against Acanthamoeba castellanii through Inhibition of the Serine Biosynthesis Pathway
Authors: Deng Yihong , Ran Wei , Man Suqin , Li Xueping , Gao Hongjian , Tang Wei , Tachibana Hiroshi , Cheng Xunjia
Published: May 2015

Acanthamoeba
sp. parasites are the causative agents of
Acanthamoeba
keratitis, fatal granulomatous amoebic encephalitis, and cutaneous infections. However, there are currently no effective drugs for these organisms. Here, we evaluated the activity of the antimalarial agent artemether against
Acanthamoeba castellanii
trophozoites and identified potential targets of this agent through a proteomic approach. Artemether exhibited
in vitro
amoebicidal activity in a time- and dose-dependent manner and induced ultrastructural modification and cell apoptosis. The iTRAQ quantitative proteomic analysis identified 707 proteins that were differentially expressed after artemether treatment. We focused on phosphoglycerate dehydrogenase and phosphoserine aminotransferase in the serine biosynthesis pathway because of their importance to the growth and proliferation of protozoan and cancer cells. The expression of these proteins in
Acanthamoeba
was validated using quantitative real-time PCR and Western blotting after artemether treatment. The changes in the expression levels of phosphoserine aminotransferase were consistent with those of phosphoglycerate dehydrogenase. Therefore, the downregulation of phosphoserine aminotransferase may be due to the downregulation of phosphoglycerate dehydrogenase. Furthermore, exogenous serine might antagonize the activity of artemether against
Acanthamoeba
trophozoites. These results indicate that the serine biosynthesis pathway is important to amoeba survival and that targeting these enzymes would improve the treatment of
Acanthamoeba
infections. Artemether may be used as a phosphoglycerate dehydrogenase inhibitor to control or block
Acanthamoeba
infections.
Four-Component Staphylococcus aureus Vaccine 4C-Staph Enhances Fcγ Receptor Expression in Neutrophils and Monocytes and Mitigates S. aureus Infection in Neutropenic Mice
Authors: Torre Antonina , Bacconi Marta , Sammicheli Chiara , Galletti Bruno , Laera Donatello , Fontana Maria Rita , Grandi Guido , De Gregorio Ennio , Bagnoli Fabio , Nuti Sandra , Bertholet Sylvie , Bensi Giuliano
Published: May 2015

Staphylococcus aureus
is a human bacterial pathogen causing a variety of diseases. The occurrence of multidrug-resistant strains of
Staphylococcus aureus
underlines the need for a vaccine. Defining immune correlates of protection may support the design of an effective vaccine. We used a murine
Staphylococcus aureus
infection model, in which bacteria were inoculated in an air pouch generated on the back of the animal. Analysis of the air-pouch content in mice immunized or not with an adjuvanted multiantigen vaccine formulation, four-component
S. aureus
vaccine (4C-Staph), prior to infection allowed us to measure bacteria, cytokines, and 4C-Staph-specific antibodies and to analyze host immune cells recruited to the infection site. Immunization with 4C-Staph resulted in accumulation of antigen-specific antibodies in the pouch and mitigated the infection. Neutrophils were the most abundant cells in the pouch, and they showed the upregulation of Fcγ receptor (FcγR) following immunization with 4C-Staph. Reduction of the infection was also obtained in mice immunized with 4C-Staph and depleted of neutrophils; these mice showed an increase in monocytes and macrophages. Upregulation of the FcγR and the presence of antigen-specific antibodies induced by immunization with 4C-Staph may contribute to increase bacterial opsonophagocytosis. Protection in neutropenic mice indicated that an effective vaccine could activate alternative protection mechanisms compensating for neutropenia, a condition often occurring in
S. aureus
-infected patients.
Host-Like Carbohydrates Promote Bloodstream Survival of Vibrio vulnificusIn Vivo
Authors: Lubin Jean-Bernard , Lewis Warren G. , Gilbert Nicole M. , Weimer Cory M. , Almagro-Moreno Salvador , Boyd E. Fidelma , Lewis Amanda L.
Published: May 2015

Sialic acids are found on all vertebrate cell surfaces and are part of a larger class of molecules known as nonulosonic acids. Many bacterial pathogens synthesize related nine-carbon backbone sugars; however, the role(s) of these non-sialic acid molecules in host-pathogen interactions is poorly understood.Vibrio vulnificusis the leading cause of seafood-related death in the United States due to its ability to quickly access the host bloodstream, which it can accomplish through gastrointestinal or wound infection. However, little is known about how this organism persists systemically. Here we demonstrate that sialic acid-like molecules are present on the lipopolysaccharide ofV. vulnificus, are required for full motility and biofilm formation, and also contribute to the organism's natural resistance to polymyxin B. Further experiments in a murine model of intravenousV. vulnificusinfection demonstrated that expression of nonulosonic acids had a striking benefit for bacterial survival during bloodstream infection and dissemination to other tissuesin vivo. In fact, levels of bacterial persistence in the blood corresponded to the overall levels of these molecules expressed byV. vulnificusisolates. Taken together, these results suggest that molecules similar to sialic acids evolved to facilitate the aquatic lifestyle ofV. vulnificusbut that their emergence also resulted in a gain of function with life-threatening potential in the human host.
Every Site Counts: Submitting Transcription Factor-Binding Site Information through the CollecTF Portal
Authors: Erill Ivan
Published: May 2015

Experimentally verified transcription factor-binding sites represent an information-rich and highly applicable data type that aptly summarizes the results of time-consuming experiments and inference processes. Currently, there is no centralized repository for this type of data, which is routinely embedded in articles and extremely hard to mine. CollecTF provides the first standardized resource for submission and deposition of these data into the NCBI RefSeq database, maximizing its accessibility and prompting the community to adopt direct submission policies.
Copper Tolerance and Characterization of a Copper-Responsive Operon,
copYAZ
, in an M1T1 Clinical Strain of Streptococcus pyogenes
Authors: Young Christie A. , Gordon Lily D. , Fang Zhong , Holder Robert C. , Reid Sean D.
Published: May 2015

Infection with
Streptococcus pyogenes
is associated with a breadth of clinical manifestations ranging from mild pharyngitis to severe necrotizing fasciitis. Elevated levels of intracellular copper are highly toxic to this bacterium, and thus, the microbe must tightly regulate the level of this metal ion by one or more mechanisms, which have, to date, not been clearly defined. In this study, we have identified two virulence mechanisms by which
S. pyogenes
protects itself against copper toxicity. We defined a set of putative genes,
copY
(for a regulator),
copA
(for a P1-type ATPase), and
copZ
(for a copper chaperone), whose expression is regulated by copper. Our results indicate that these genes are highly conserved among a range of clinical
S. pyogenes
isolates. The
copY
,
copA
, and
copZ
genes are induced by copper and are transcribed as a single unit. Heterologous expression assays revealed that
S. pyogenes
CopA can confer copper tolerance in a copper-sensitive
Escherichia coli
mutant by preventing the accumulation of toxic levels of copper, a finding that is consistent with a role for CopA in copper export. Evaluation of the effect of copper stress on
S. pyogenes
in a planktonic or biofilm state revealed that biofilms may aid in protection during initial exposure to copper. However, copper stress appears to prevent the shift from the planktonic to the biofilm state. Therefore, our results indicate that
S. pyogenes
may use several virulence mechanisms, including altered gene expression and a transition to and from planktonic and biofilm states, to promote survival during copper stress.

IMPORTANCE
Bacterial pathogens encounter multiple stressors at the host-pathogen interface. This study evaluates a virulence mechanism(s) utilized by
S. pyogenes
to combat copper at sites of infection. A better understanding of pathogen tolerance to stressors such as copper is necessary to determine how host-pathogen interactions impact bacterial survival during infections. These insights may lead to the identification of novel therapeutic targets that can be used to address antibiotic resistance.
Critical Components of the Conjugation Machinery of the Integrative and Conjugative Element ICE
Bs1
of Bacillus subtilis
Authors: Leonetti Cori T. , Hamada Matt A. , Laurer Stephanie J. , Broulidakis Matthew P. , Swerdlow Kyle J. , Lee Catherine A. , Grossman Alan D. , Berkmen Melanie B.
Published: May 2015

Conjugation, or mating, plays a profound role in bacterial evolution by spreading genes that allow bacteria to adapt to and colonize new niches. ICE
Bs1
, an integrative and conjugative element of
Bacillus subtilis
, can transfer itself and mobilize resident plasmids. DNA transfer is mediated by a type IV secretion system (T4SS). Characterized components of the ICE
Bs1
T4SS include the conserved VirB4-like ATPase ConE, the bifunctional cell wall hydrolase CwlT, and the presumed VirD4-like coupling protein ConQ. A fusion of ConE to green fluorescent protein (GFP) localizes to the membrane preferentially at the cell poles. One or more ICE
Bs1
proteins are required for ConE's localization at the membrane, as ConE lacks predicted transmembrane segments and ConE-GFP is found dispersed throughout the cytoplasm in cells lacking ICE
Bs1
. Here, we analyzed five ICE
Bs1
genes to determine if they are required for DNA transfer and/or ConE-GFP localization. We found that
conB
,
conC
,
conD
, and
conG
, but not
yddF
, are required for both ICE
Bs1
transfer and plasmid mobilization. All four required genes encode predicted integral membrane proteins.
conB
and, to some extent,
conD
were required for localization of ConE-GFP to the membrane. Using an adenylate cyclase-based bacterial two-hybrid system, we found that ConE interacts with ConB. We propose a model in which the ICE
Bs1
conjugation machinery is composed of ConB, ConC, ConD, ConE, ConG, CwlT, ConQ, and possibly other ICE
Bs1
proteins, and that ConB interacts with ConE, helping to recruit and/or maintain ConE at the membrane.

IMPORTANCE
Conjugation is a major form of horizontal gene transfer and has played a profound role in bacterial evolution by moving genes, including those involved in antibiotic resistance, metabolism, symbiosis, and infectious disease. During conjugation, DNA is transferred from cell to cell through the conjugation machinery, a type of secretion system. Relatively little is known about the conjugation machinery of Gram-positive bacteria. Here, we analyzed five genes of the integrative and conjugative element ICE
Bs1
of
Bacillus subtilis
. Our research identifies four new components of the ICE
Bs1
conjugation machinery (ConB, ConC, ConD, and ConG) and shows an interaction between ConB and ConE that is required for ConE to associate with the cell membrane.